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    Cell and Molecular Biology
    The effect of exosomal miR-1260a derived from human umbilical mesenchymal stem cells on apoptosis of granulosa cells in PCOS patients
    ZHAO Yuanyuan, WU Xiaohua
    2023, 51 (4):  337-343.  doi: 10.11958/20221070
    Abstract ( 597 )   HTML ( 10 )   PDF (3916KB) ( 952 )  

    Objective To explore the effect of exosomal miR-1260a derived from human umbilical mesenchymal stem cells (hUC-MSCs) on apoptosis of granulosa cells (GCs) in polycystic ovary syndrome (PCOS) by targeting CASP8. Methods hUC-MSCs were obtained from the umbilical cord of healthy full-term fetus by tissue explants adherent method. The surface markers CD73 and CD90 were detected by flow cytometry. The hUC-MSCs- exosomes (exos) were extracted from the supernatant of hUC-MSCs, and the morphology was observed by transmission electron microscopy (TEM). The surface heat shock protein (HSP70) and tumor susceptibility gene 101 (TSG101) were detected by Western blot assay. Confocal microscope was used to observe whether PKH67 labeled hUC-MSCs-exos could be absorbed by GCs. Western blot assay was used to assess the protective effect of hUC-MSCs-exos on PCOS GCs. Clinical samples of GSE69909 were used for bioinformatics analysis. The differentially expressed miRNAs in hUC-MSCs-exos were obtained and verified by qPCR. miR-1260a mimics and negative control (NC) were synthesized and then transfected into KGN cells. Subsequently, cell apoptosis was measured by flow cytometry. Target genes of miR-1260a were predicted using Target Scan Human 7.2 database and verified by a dual-luciferase reporter gene assay. Expression levels of miR-1260a and CASP8 before and after uptake of hUC-MSCs-exos by PCOS GCs were detected by qPCR. Results The isolated cells accorded with the characteristics of hUC-MSCs. hUC-MSCs-exos were circular or elliptical membranous vesicle with diameter ranged from 30-150 nm and expressed the typical exosome markers HSP70 and TSG101. Confocal microscopy results showed that hUC-MSCs-exos could be absorbed by GCs. Compared with the control group, the expression of Cleaved-Caspase-8, Cleaved-Caspase-3 and Bax were significantly down-regulated and the expression of Bcl-2 was significantly up-regulated in the hUC-MSCs supernatant treated and the hUC-MSCs-exos treated groups (P<0.05). After GSE69909 database screening and qPCR verification, it was found that miR-1260a was enriched in hUC-MSCs-exos, and the overexpression of miR-1260a significantly inhibited the apoptosis of KGN cells (P<0.05). A dual-luciferase reporter gene assay showed that CASP8 was a direct target of miR-1260a. Moreover, the uptake of hUC-MSCs-exos by PCOS GCs increased the expression of miR-1260a and decreased the expression of CASP8 (P<0.05). Conclusion Our study reveals that exosomal miR-1260a derived from hUC-MSCs inhibits apoptosis of GCs in PCOS by targeting CASP8.

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    Molecular mechanism of hyperoside inhibiting the biological behavior of nasopharyngeal carcinoma cells
    GE Yuan, MA Zuopeng, WANG Yanhua
    2023, 51 (4):  344-350.  doi: 10.11958/20220805
    Abstract ( 313 )   HTML ( 7 )   PDF (1831KB) ( 937 )  

    Objective To investigate the effect and mechanism of hyperoside on the biological behavior such as cell proliferation, migration and invasion of nasopharyngeal carcinoma cells. Methods SUNE1 cells were incubated with hyperin medium of 10 mg/L, 20 mg/L and 40 mg/L, and cells were divided into the low, the medium and the high dose groups. The lncRNA PAX8-AS1 overexpression or knockdown plasmid and miR-494-3p mimic were transfected into 40 mg/L hypericin-treated SUNE1 cells respectively. The activity, clonal formation, migration and invasion of SUNE1 cells were detected by CCK-8 assays, plate cloning experiment and Transwell assays. Western blot assay was used to analyze the expression of matrix metalloproteinase (MMP) -2 and MMP-9 proteins. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of long non-coding RNA (lncRNA) paired-box gene 8 antisense RNA 1 (PAX8-AS1) and miR-494-3p. Dual luciferase report experiment and RT-qPCR were used to determine the targeting relationship between lncRNA PAX8-AS1 and miR-494-3p. Results Low, medium and high dose-hyperoside significantly reduced the cell viability, migration number and invasion number of SUNE1 cells, and down-regulated expression levels of MMP-2 protein, MMP-9 protein and miR-494-3p, and up-regulated the expression level of lncRNA PAX8-AS1 (P<0.05). lncRNA PAX8-AS1 negatively regulated miR-494-3p expression. lncRNA PAX8-AS1 overexpression significantly enhanced inhibitory effects of 40 mg/L hyperoside on SUNE1 cell viability, migration and invasion, and MMP-2 and MMP-9 protein expression (P<0.05). Knockdown of lncRNA PAX8-AS1 and overexpression of miR-494-3p significantly reduced inhibitory effects of 40 mg/L hyperoside on SUNE1 cell viability, migration, invasion and MMP-2 and MMP-9 protein expression (P<0.05). Conclusion Hyperoside may inhibit the proliferation and migration, and invasion of nasopharyngeal carcinoma cells by up-regulating the lncRNA PAX8-AS1 /miR-494-3p axis.

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    Study on the expression of pyrolysis-related factors in the decidua tissue of patients with early abortion after IVF/ICSI assisted pregnancy
    CHE Xiaoqun, WANG Lilian, WANG Xiao, XU Lihua, MA Tianzhong, JING Xia, CHEN Bi
    2023, 51 (4):  351-354.  doi: 10.11958/20220918
    Abstract ( 318 )   HTML ( 3 )   PDF (804KB) ( 990 )  

    Objective To investigate the expression and effect of pyrodeath related factors in decidual tissue of in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) assisted early postpregnancy abortion. Methods A total of 102 patients with early abortion after IVF/ICSI assisted pregnancy were selected as the research group, and 100 patients who underwent artificial abortion due to unexpected pregnancy during the same period were selected as the control group. Samples of decidua tissue were collected. Quantitative real-time PCR (qRT-PCR) method and Western blot assay were used to detect expression levels of Nod-like receptor pyrin domain 3 (NLRP3), interleukin-1β (IL-1β), Cysteinyl aspartate specific proteinase 1 (Caspase1), gasdermin D (GSDMD) mRNA and protein in decidua tissue. TUNEL method was used to detect the cell pyroptosis in decidua tissue. Pearson method was used to analyze the correlation between pyroptosis rate and the protein levels of NLRP3, IL-1β, Caspase1 and GSDMD in decidua. Results Compared with the control group, expression levels of NLRP3, IL-1β, Caspase1, GSDMD mRNA and protein, the cell pyroptosis rate in decidua tissues were significantly increased in the research group (P<0.05). There was a positive correlation between cell pyroptosis rate in decidua tissue and expression levels of NLRP3, IL-1β, Caspase1 and GSDMD in the research group (r=0.429, 0.474, 0.498, 0.540, all P<0.05). Conclusion The expression levels of pyrolysis-related factors NLRP3, IL-1β, Caspase1 and GSDMD in decidua tissue are increased in patients with early abortion after IVF/ICSI assisted pregnancy, and which are related to the cell pyroptosis of decidua tissue.

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    Inhibitory effect and mechanism of metformin on diffuse large B-cell lymphoma cells
    TAN Chunlian, LI Xiaohong, XIA Guodong, ZHANG Zhihong, LI Xiaoming
    2023, 51 (4):  355-359.  doi: 10.11958/20220684
    Abstract ( 622 )   HTML ( 6 )   PDF (1250KB) ( 929 )  

    Objective To investigate the inhibitory effect and related mechanism of metformin on human diffuse large B-cell lymphoma (DLBCL) U2932 cells. Methods U2932 cells cultured in vitro were treated with different concentrations of metformin, and cells were divided into the control group (0 mmol/L) and the metformin experimental groups (5, 10, 20 and 40 mmol/L). Cells were cultured for 24 h, 48 h and 72 h, respectively. Changes of cell proliferation activity were detected by CCK-8 assay. Cell cycle distribution and apoptosis were detected by flow cytometry. Expression levels of AMPK, CyclinD1, autophagy and apoptosis pathway-related proteins were detected by Western blot assay. Results The proliferation activity of U2932 cells treated with metformin was significantly lower than that of the control group (P<0.05). Compared with the control group, the percentage of cells in G0/G1 phase treated with 5, 10, 20 mmol/L metformin and the apoptosis rate of U2932 cells treated with 20 mmol/L metformin for 24 h, 48 h and 72 h were significantly increased (P<0.05). Western blot assay showed that 5, 10, 20 mmol/L metformin could up-regulate the expression of p-AMPKα, P53, Bax, Cleaved Caspase-3, LC3B proteins, but Beclin1 only partially increased, and down-regulate the expression levels of Bcl-2, p-AKT, CyclinD1 proteins in U2932 cells (P<0.05). Conclusion Metformin can activate AMPK, apoptosis and autophagy pathways, inhibit the proliferation of U2932 cells, and promote their apoptosis.

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    Experimental Research
    Study on the mechanism of let-7e-5p regulating Fas/FasL to affect the pathogenesis of allergic rhinitis in mice
    TANG Qiaofei, CAO Jianqiu, ZHANG Shuang
    2023, 51 (4):  360-365.  doi: 10.11958/20221110
    Abstract ( 290 )   HTML ( 7 )   PDF (1151KB) ( 936 )  

    Objective To investigate the effect and mechanism of let-7e-5p on allergic rhinitis in mice. Methods Eighteen BALB/c mice were randomly divided into the control group, the allergic rhinitis (AR) group, the let-7e-5p agonist negative control (AR+agomir-NC) group, the let-7e-5p agonist (AR+agomir) group, the let-7e-5p inhibitor negative control (AR+antagomir-NC) group and the let-7e-5p inhibitor (AR+antagomir) group. Except the control group, the other groups were sensitized with ovalbumin (OVA) to establish AR model. The AR+agomir group and the AR+antagomir group were treated with let-7e-5p agomir or let-7e-5p antagomir at the same time for intervention. After the last sensitization, the allergic rhinitis of mice were scored. HE staining was used to observe nasal mucosa. The number of eosinophils was counted. Real-time PCR was used to detect mRNA expression levels of let-7e-5p, Fas and FasL, and Western blot assay was used to detect protein expression levels of Fas and FasL. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum levels of immunoglobulin E (IgE), specific immunoglobulin E (sIgE), interferon gamma (IFN-γ), interleukin (IL)-12, IL-4 and IL-13. The targeting relationship between let-7e-5p and Fas and FasL were analyzed by dual luciferase assay. Results Compared with the control group, the allergic rhinitis score was increased in the AR group, nasal mucosa hyperplasia was obvious and the number of eosinophils was increased. The expression levels of let-7e-5p and IFN-γ and IL-12 were decreased, while the expression of Fas and FasL and levels of IgE, sIgE, IL-4 and IL-13 were increased (all P<0.05). Compared with the AR+agomir-NC group, the allergic rhinitis score was decreased in the AR+agomir group, the nasal mucosa structure was clear and the number of eosinophils was reduced. The expression of let-7e-5p and levels of IFN-γ and IL-12 were increased, while the expression of Fas and FasL and levels of IgE, sIgE, IL-4 and IL-13 were decreased (all P<0.05). Compared with the AR+antagomir-NC group, the allergic rhinitis score was increased in the AR+antagomir group, the nasal mucosa was significantly thickened, and the number of eosinophils was increased. The expression of let-7e-5p was decreased, and the expression of Fas, FasL and levels of IgE, sIgE, IL-4 and IL-13 were increased (all P<0.05). There were no significant differences in IL-12 and IFN-γ between the AR+antagomir-NC group and the AR+antagomir group (P>0.05). Compared with the NC+3'UTR-WT group, luciferase activity was decreased in the let-7e-5p agomir+3'UTR-WT group (P<0.05). Conclusion Let-7e-5p may affect the balance of T helper type 1 (Th1)/ T helper type 2 (Th2) cells by targeting Fas/FasL, and thus participate in the progression of AR in mice.

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    The role of sigma-1 receptor in endoplasmic reticulum stress in neuropathic pain rats
    MO Shaoe, LIU Kepeng, FU Wen, XIE Chouqin, YU Dongling, CHEN Shi, LAN Yuyan
    2023, 51 (4):  366-370.  doi: 10.11958/20221491
    Abstract ( 469 )   HTML ( 5 )   PDF (1298KB) ( 958 )  

    Objective To investigate the role of sigma-1 receptor (sig-1R) in endoplasmic reticulum stress in neuropathic pain (NP) rats. Methods Twenty-four SPF male SD rats with successful sheath tube were divided into the sham operation group (group S), the neuropathic pain model group (group C) and the BD1047 intervention group (group B) according to random number table, with 8 rats in each group. Chronic crush injury of sciatic nerve was used to establish NP model. On the 4th, 5th and 6th day after operation, normal saline 20 μL was injected intrathecally in the group S and the group C, and BD1047 (120 μmol/L) 20 μL was injected intrathecally in the group B once a day. The mechanical foot withdrawal response threshold (MWT) of the surgical side foot was measured on the first day before operation and on the first, 3rd, 5th and 7th day after operation. Western blot assay and immunofluorescence staining were used to detect the expression levels of sig-1R, glucose-regulated protein (BIP), activating transcription factor 4 (ATF4) and C/EBP-homologous protein (CHOP) in dorsal root neurons (DRG). HE staining was used to observe the morphological size and pathological changes of DRG. Changes of endoplasmic reticulum in DRG were observed by scanning electron microscopy. Results Compared with the group S, intraoperative parapodum MWT was significantly decreased (P<0.05), and pathological injury and endoplasmic reticulum destruction of DRG were significantly aggravated. Expressions of sig-1R, BIP, ATF4 and CHOP were up-regulated in the group C after CCI (P<0.05). Compared with the group C, the MWT was increased on the 5th and 7th day after CCI in the group B (P<0.05), and the expressions of sig-1R, BIP, ATF4 and CHOP were down-regulated (P<0.05). The pathological injury of DRG was improved. Conclusion sig-1R are involved in the neuropathic pain in rats, and the mechanism may be related to the inhibition of endoplasmic reticulum stress in dorsal root neurons.

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    Protective effect of Klotho protein on acute renal injury and fibrosis in rats with ischemia-reperfusion
    LIU Zhufeng, WANG Wenhong, FAN Shuying, LIU Yan, LIU Tao, WANG Xin, WU Xia
    2023, 51 (4):  371-375.  doi: 10.11958/20221063
    Abstract ( 467 )   HTML ( 4 )   PDF (1179KB) ( 921 )  

    Objective To investigate the role of Klotho protein in acute renal injury and fibrosis induced by ischemia-reperfusion (I/R) in rats. Methods Sixty healthy SD rats were randomly divided into the sham operation group (sham group), the renal ischemia-reperfusion group (I/R group) and the Klotho protein intervention group (I/R+Kl group), with 20 rats in each group. In the I/R group, the acute renal injury model of I/R was established by clamping the right renal pedicle for 40 min. In the sham group, the right renal artery was dissociated, but the right renal pedicle was not clamped. After exposure for 40 minutes, the abdominal cavity was sutured. The operation method of the I/R+Kl group was the same as that of the I/R group. Kiotho protein (0.01 mg/kg) was injected intraperitoneally. The blood and renal tissue samples were collected on the 1st, 7th, 14th and 28th days after successful modeling in the three groups, and blood creatinine (Scr) and blood urea nitrogen (BUN) were measured respectively. HE staining was used to observe the damage of renal tissue. Masson staining was used to observe the renal fibrosis. ELISA was used to detect levels of Klotho protein in blood and renal tissue. Western blot assay was used to detect the expression of Klotho protein in renal tissue. Results Compared with the sham group, levels of SCR and BUN were increased in the I/R group and the I/R+Kl group (P<0.05). Compared with the I/R+Kl group, serum levels of SCR and BUN were increased in different time points in the I/R group (P<0.05). HE staining showed extensive acute necrosis of renal tubules in the I/R group, and the injury was mild in the I/R+Kl group. Masson staining showed that mild fibrosis occurred in the I/R group on the 14th day, and the degree of fibrosis was more significant on the 28th day, while the degree of fibrosis was very mild in the I/R+Kl group (P < 0.05). ELISA results showed that compared with the Sham group at day 1, 14 and 28, levels of Klotho in blood and renal tissue decreased significantly in the I/R+Kl group and the I/R group, and the level of Klotho decreased in the I/R group than that of the I/R+Kl group (P<0.05). Western blot results showed that compared with the Sham group, Klotho levels were decreased at all time points in the I/R group and the I/R+Kl group. while Klotho levels were higher in the I/R+Kl group than those in the I/R group (P<0.05). Conclusion Klotho expression is down-regulated during renal I/R injury. Exogenous supplementation of Klotho protein can alleviate acute renal injury and fibrosis in rats with renal I/R injury and play a role in renal protection.

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    Effects and mechanism of hyperuricemia on spermatogenesis and sperm quality in mice
    JIANG Xiaocui, TIAN Daizhi, ZHAO Min, GONG Jian, YU He, JIANG Xingyu, XIAO Min
    2023, 51 (4):  376-381.  doi: 10.11958/20221083
    Abstract ( 497 )   HTML ( 6 )   PDF (2973KB) ( 1466 )  

    Objective To investigate the mechanism of hyperuricemia (HUA) inducing testicular cell apoptosis and reducing spermatogenesis and sperm quality in mice through oxidative damage. Methods Thirty-six Kunming male mice were divided into 6 groups by random number method: the potassium oxyazinate groups for 1 d, 7 d and 14 d, and the control groups for 1 d, 7 d and 14 d. The potassium oxyazinate group was intraperitoneally injected with potassium oxyazinate suspension 600 mg/(kg·d). Serum levels of uric acid (UA), creatinine (Cre), urea nitrogen (BUN), alanine aminotransferase (ALT), aspartate aminotransferase (AST), xanthine oxidase (XO), and superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) in testicular tissue were detected by biochemical method. Eosin-hematoxylin (HE) staining was used to observe histopathological changes of liver, kidney and testis, and the spermatogenic function of testicular HE sections was scored. Automatic sperm analyzer detected sperm density and motility rate. The expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and Caspase-3 protein in testicular tissue were detected by Western blotting. Results Compared with the control group, UA was significantly increased in the potassium oxazinate 1 d group (P<0.01). The levels of UA, BUN, AST and XO were significantly increased in the potassium oxazinate 7 d group (P<0.01). The levels of UA, BUN and AST were significantly increased in the potassium oxazinate 14 d group (P<0.01). The pathological changes in liver and kidney tissue were observed in the potassium oxazinate 7 d and 14 d groups, and the lesion was milder in the potassium oxazinate 7 d groug than that of the potassium oxazinate 14 d group. Compared with the control 7 d group, there were obvious pathological changes and significantly decreased spermatogenic function in the potassium oxazinate 7 d group (P<0.01). The content of MDA was significantly increased (P<0.05), while the activities of SOD and CAT were significantly decreased (P<0.05). The protein expression levels of Bax and Caspase-3 were significantly increased (P<0.05), the ratio of Bax to Bcl-2 was also increased (P<0.05), and the protein expression level of Bcl-2 was significantly decreased (P<0.05). Conclusion The spermatogenic function and sperm quality are decreased in HUA mice, which may be caused by testicular oxidative damage induced apoptosis.

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    Effects of tetramethylpyrazine on analgesia and neuronal damage in migraine rats by regulating SIRT1/AMPK/PGC1α signaling pathway
    HU Bin, WANG Dabin, GUO Mao
    2023, 51 (4):  382-386.  doi: 10.11958/20220873
    Abstract ( 570 )   HTML ( 7 )   PDF (999KB) ( 956 )  

    Objective To explore the effects of tetramethylpyrazine (TMP) on analgesia and neuronal injury protection in migraine rats by regulating silent mating type information regulation 2 homolog 1 (SIRT1)/AMP activated protein kinase (AMPK)/peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) signaling pathway. Methods The migraine rat model was established by nitroglycerin induction. After successful modeling, rats were randomly divided into the model (M) group, the TMP low dose (TMP-L) group (50 mg/kg), the TMP medium dose (TMP-M) group (100 mg/kg), the TMP high dose (TMP-M) group (200 mg/kg) and the TMP (200 mg/kg) + SIRT1 inhibitor (EX527, 5 mg/kg) group, 10 rats in each group. Another 10 rats were regarded as the normal control (NC) group. Rats were continuously gavaged for 2 weeks. Twenty-four hours after the end of the administration, the times of scratching head scratching and cage climbing of rats within 30 minutes were recorded in each group, and the behavioral score was carried out. The pain threshold for mechanical stimulation and thermal stimulation were determined. ELISA method was applied to measure serum levels of nitric oxide (NO), interleukin-6 (IL-6), interleukin-1β (IL-1β), and 5-hydroxytryptamine (5-HT), norepinephrine (NE) and dopamine (DA) in brain tissue. TUNEL staining was applied to observe neuronal apoptosis in brain tissue. Western blot assay was applied to measure the protein expression levels of SIRT1, AMPK, p-AMPK and PGC1α protein in brain tissue. Results Compared with the NC group, the behavioral score, serum levels of NO, IL-6 and IL-1β, and neuron apoptosis rate were significantly increased in the M group (P<0.05). The pain threshold of mechanical stimulation was significantly reduced, and the latency of thermal stimulation was significantly shortened (P<0.05). The levels of 5-HT, NE and DA in brain tissue, the ratio of p-AMPK/AMPK, and the protein expressions of SIRT1 and PGC1α were significantly decreased (P<0.05). Compared with the M group, the behavioral score, the serum levels of NO, IL-6 and IL-1β, and neuron apoptosis rate were significantly decreased in the TMP groups (P<0.05). The pain threshold of mechanical stimulation was significantly increased, and the latency of thermal stimulation was significantly prolonged (P<0.05). The levels of 5-HT, NE and DA in brain tissue, the ratio of p-AMPK/AMPK, and the protein expressions of SIRT1 and PGC1α were significantly increased (P<0.05). Compared with the TMP-H group, TMP+EX527 group showed that it significantly reversed the effect of TMP on migraine rats. Conclusion TMP may improve neuronal damage by regulating the expression of SIRT1/AMPK/PGC1α signaling pathway and exert analgesic effect on migraine rats.

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    Study on the mechanism of glial maturation factor-β induced inflammatory response of retinal Müller cells in diabetes rats
    LUO Ying, SHAN Wei, ZHANG Qiao
    2023, 51 (4):  387-391.  doi: 10.11958/20221106
    Abstract ( 385 )   HTML ( 4 )   PDF (1261KB) ( 935 )  

    Objective To study the activation and its related mechanisms of glial maturation factor-β(GMFB) on retinal MüLler cells in diabetes rats. Methods Sixty SPF grade male SD rats were divided into the streptozotocin (STZ) group, the STZ+AAV-GMFB group and the STZ+AAV-GMFB+K252a (TrkB inhibitor) group with 15 rats in each group. Another 15 normal rats were taken as the CON group. Rats in the STZ+AAV-GMFB group and the STZ+AAV-GMFB+K252a group were injected with AAV-GMFB adenovirus vector 5 μL in vitreous cavity 8 weeks after modeling. The STZ+AAV-GMFB+K252a group received intraperitoneal injection of K252a (25 μg·kg-1·d-1). After 12 weeks, the expression of GMFB in Müller cells was detected by immunofluorescence, the expression of glial fibrillary acidic protein (GFAP) in retina was detected by immunohistochemistry, and the retinal inflammatory factor tumor necrosis factor- α (TNF-α), interleukin (IL)-1β and IL-6 were detected by ELISA. The relative expression levels of GMFB, brain derived neurotrophic factor (BDNF) and its tyrosine kinase receptor B (p-TrkB) were detected by Western blot assay. HE staining was used to detect the pathological changes of retina. Results GMFB was expressed in Müller cells. Compared with the CON group, the expression levels of GMFB, GFAP and TNF-α, IL-1β, IL-6 levels increased, the number of retinal ganglion cells (RGC) and the expression levels of BDNF, p-TrkB decreased in the STZ group. Compared with the STZ group, the expression levels of GMFB, GFAP and TNF-α, IL-1β and IL-6 levels decreased, the expression of BDNF, p-TrkB and the number of RGC were increased in the STZ+AAV-GMFB group. However, K252a can largely reverse the protective effect of AAV-GMFB. Conclusion The expression of GMFB in the retina of rats with diabetes is increased, which can induce the activation of Müller cells and aggravate the inflammatory response. This effect is related to the inhibition of BDNF/TrkB signal pathway.

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    Original article
    The influence of posterior lumbar interbody fusion on paraspinal muscle injury
    ZHANG Xiao, SHANG Liang, RUAN Zhi
    2023, 51 (4):  392-394.  doi: 10.11958/20221658
    Abstract ( 305 )   HTML ( 5 )   PDF (747KB) ( 931 )  

    Objective To evaluate paraspinal muscle injury after posterior midline lumbar interbody fusion. Methods Fifty-six patients with lateral recess lumbar spinal stenosis who received PLIF were prospectively included. Visual analog scale (VAS) and Oswestry Disability Index (ODI) were recorded before surgery and 3 months after surgery. Surface electromyography (sEMG) was used to measure the root mean square (RMS) and median frequency (MF) of paravertebral muscle in the three stages of lift test. Serum levels of creatine kinase (CK) and C-reactive protein (CRP) were detected before surgery and at 1, 3 and 7 days after surgery. Results The VAS score and ODI score were significantly lower in patients 3 months after surgery than those before surgery (P<0.05). In test stage 1 and 2, the level of paraspinal muscle RMS was significantly lower 3 months after surgery than that before surgery, and in test stage 3, the postoperative paraspinal muscle MF value was significantly higher than that before surgery (P<0.05). Serum CK level was significantly higher on the 1st and 3rd day after surgery than that before surgery, and which reached the peak on the 1st day after surgery (P<0.05). Serum CK level basically recovered to the preoperative level on the 7th day after surgery, and CRP level was higher at all time points after surgery than that before surgery, and reached the peak on the 3rd day after surgery, and decreased on the 7th day after surgery (P<0.05). Conclusion sEMG reveals that posterior lumbar interbody fusion causes some paraspinal muscle tissue injury, which may be the primary cause of postoperative paraspinal muscle function decline and postoperative low back pain.

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    Clinical Research
    Correlation between homocysteine and left ventricular myocardial fibrosis in hypertensive patients
    ZHENG Yue, MA Yunting, ZHAO Xiaoying, ZHAO Xinxiang
    2023, 51 (4):  395-399.  doi: 10.11958/20221111
    Abstract ( 327 )   HTML ( 5 )   PDF (866KB) ( 954 )  

    Objective To investigate the relationship between levels of homocysteine (Hcy) and left ventricular myocardial fibrosis measured by late gadolinium enhancement (CMR-LGE) in hypertensive patients. Methods Eighty-seven hypertensive patients who underwent cardiac magnetic resonance (CMR) examination were selected. According to Hcy level, patients were divided into the normal Hcy group (Hcy≤15 μmol/L, 40 cases) and the high Hcy group (Hcy>15 μmol/L, 47 cases). The clinical data of all patients were completed between the two groups. Spearman rank correlation analysis was used to analyze the correlation between Hcy and late gadolinium enhancement volume percentage (LGE%), and the correlation between Hcy and other cardiac parameters. Results The left atrial diameter (LAD), left ventricular end-diastolic diameter (LVEDD), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular mass (LVM), left ventricular mass index (LVMI) and LGE% were significantly higher in the high Hcy group than those in the normal Hcy group. The left ventricular ejection fraction (LVEF) was significantly lower in the high Hcy group than that of the normal Hcy group (P<0.05). Spearman correlation analysis showed that Hcy was positively correlated with LAD, LVEDD, LVEDV, LVESV, LVM, LVMI and LGE% (rs were 0.356, 0.332, 0.288, 0.383, 0.338, 0.325, 0.391, respectively, all P<0.01). Hcy was negatively correlated with LVEF (rs=-0.412, P<0.01). Conclusion Hypertensive patients with high Hcy level have severe left ventricular myocardial fibrosis, resulting in more serious left heart structure and dysfunction.

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    Expression and clinical significance of CD163 in hepatic fibrosis with biliary atresia
    LIU Zhijie, ZHENG Qipeng, XU Xiaodan, WANG Qiong, LI Mengdi, ZHANG Cong, WANG Zhiru, ZHAN Jianghua
    2023, 51 (4):  400-403.  doi: 10.11958/20221539
    Abstract ( 495 )   HTML ( 5 )   PDF (1386KB) ( 943 )  

    Objective To investigate the expression of CD163 in children with biliary atresia (BA) and its relationship with liver fibrosis. Methods A total of 30 children with BA (the BA group, 14 patients with liver fibrosis grade Ⅰ-Ⅱ, 16 patients with liver fibrosis grade Ⅲ-Ⅳ) and 10 children with choledochal cyst (the CC group) were collected. Samples of liver tissue and blood were obtained from the 2 groups. The expression levels of CD163 in liver tissue and blood of the two groups were detected by immunohistochemistry and flow cytometry. The expression of sCD163 was detected by enzyme-linked immunosorbent assay (ELISA), and the value of sCD163 in evaluating BA liver fibrosis was analyzed by receiver operating characteristic curve (ROC). Results Immunohistochemical staining and flow cytometry indicated that CD163 expression levels in liver tissue and blood were significantly higher in the BA group than those in the CC group (P<0.01), and which increased with the increased severity of liver fibrosis (P<0.01). The expression of sCD163 also showed the same change. sCD163 predicted BA with an area under the ROC curve of 0.927, a best threshold value of 8.323 μg/L, a sensitivity of 86.67% and a specificity of 100%. Conclusion The expression of CD163 is up-regulated in children with BA, which accelerates the process of liver fibrosis. sCD163 can accurately reflect the severity of liver fibrosis in children with BA.

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    Risk factors of appendiceal orifice inflammatory in patients with ulcerative colitis and prognosis analysis
    SHI Ganggang, ZHENG Bing, HAN Mei
    2023, 51 (4):  404-408.  doi: 10.11958/20221581
    Abstract ( 536 )   HTML ( 4 )   PDF (1108KB) ( 951 )  

    Objective To analyze the risk factors of appendiceal orifice inflammation (AOI) in patients with ulcerative colitis (UC) and the influence of AOI on the prognosis of UC. Methods A total of 288 UC patients were enrolled, of them 105 were AOI positive and 183 were AOI negative. The medical records of UC patients who underwent colonoscopy and treatment in the department of colorectal surgery were collected, and data including gender, age, medical history, colonoscopic lesion involvement and Mayo endoscopic subscore, microscopic pathological features and treatment. All patients were divided into two groups according to AOI. The demographics and endoscopic features of the two groups were analyzed. Logistic regression was used to analyze risk factors of AOI in UC patients. Kendall grade correlation was used to analyze the correlation between AOI inflammatory grade and rectal inflammatory grade in AOI-positive UC patients. Results There were no significant differences in age, gender and clinical classification between the two groups. There were significant differences in the scope of lesion involved and Mayo endoscopic subscore between the two groups (P<0.05). Multivariate Logistic regression analysis showed that wide lesion involvement and higher Mayo endoscopic subscore were independent risk factors for AOI (P<0.05). In AOI-positive UC patients, the grade of AOI pathological inflammation was positively correlated with the grade of rectal pathological inflammation (rs=0.512, P<0.05). After treatment, most patients achieved remission, and there was no significant difference in the therapeutic effect between AOI positive and AOI negative patients. According to the treatment methods, AOI positive patients with proctitis were divided into the suppository drug group and the suppository combined with oral drug group, and the remission rate was higher in the suppository combined with oral drug group than that of the suppository drug group (P<0.05). Conclusion The wider the scope of endoscopic lesion involvement and the higher the Mayo endoscopic subscore in UC patients, the more likely AOI positive is. The evaluation of AOI under the colonoscopy is beneficial to better guide the treatment of UC patients.

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    Effects and clinical efficacy of breviscapine on inflammatory factors and coagulation function in patients with septic acute lung injury
    ZHOU Ying, WANG Yaning, WU Shuang, KE Huijuan, SUN Pengfei
    2023, 51 (4):  409-412.  doi: 10.11958/20221250
    Abstract ( 400 )   HTML ( 5 )   PDF (764KB) ( 956 )  

    Objective To explore effects and clinical efficacy of breviscapine on inflammatory factors and coagulation function in patients with septic acute lung injury (ALI). Methods A total 84 patients with ALI caused by sepsis were assigned to the control group, the low-dose breviscapine group (low-dose group) and the high-dose breviscapine group (high-dose group) according to random digital table, with 28 cases in each group. The control group was treated with conventional Western medicine. The low-dose group was additionally given breviscapine injection (20 mg, one time/d) on the basis of the control group, and the high-dose group was additionally given breviscapine injection (40 mg, one time/d) on the basis of the control group. The three groups were treated for 2 weeks. The clinical efficacy were evaluated in the three groups. Traditional Chinese medicine (TCM) syndrome scores, oxygenation index (PaO2/FiO2), interleukin (IL) -6 and tumor necrosis factor-α (TNF-α) levels, prothrombin time (PT), activated partial thromboplastin time (APTT) and platelet count were compared before and after two weeks of treatment between the three groups. Drug-related adverse events during treatment were recorded in each group. Results The clinical efficacy and total effective rate were better in the low-dose group and the high-dose group compared with those of the control group (P<0.05). After treatment, the TCM syndrome score and levels of IL-6 and TNF-α were decreased than those before treatment in the three groups (P<0.05), and the control group, the low-dose group and the high-dose group were decreased successively. After treatment, PT and APTT were prolonged compared with those before treatment in the three groups, and the control group, the low-dose group and the high-dose group were prolonged successively (P<0.05). After treatment, PaO2/FiO2 and platelet count were increased compared with those before treatment in the three groups, and the control group, the low-dose group and the high-dose group were increased successively (P<0.05). There were no severe adverse drug events during the treatment in the three groups. Conclusion Breviscapine can significantly improve the coagulation dysfunction in sepsis, reduce the abnormal consumption of platelets, inhibit inflammatory response, and has a relatively definite clinical effect on patients with sepsis-induced acute lung injury.

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    Serum levels of ET-1 and SDC-1 in children with nephrotic syndrome and their correlation with hormone therapy response
    LIU Qing, ZHU Jun, TANG Guoying, JIANG Pingying
    2023, 51 (4):  413-417.  doi: 10.11958/20221637
    Abstract ( 350 )   HTML ( 4 )   PDF (765KB) ( 932 )  

    Objective To investigate the correlation between serum levels of endothelin-1 (ET-1) and polyligand proteoglynan-1 (SDC-1) with the responsiveness to hormone therapy in children with nephrotic syndrome (NS). Methods A total of 157 children with NS who were expected to receive hormone therapy and 67 healthy children matching the age and sex with NS at the same time were selected (the control group). Clinical data were collected and serum levels of ET-1, SDC-1, 24 h urine protein, urea nitrogen (BUN), serum creatinine (Scr) and other laboratory indexes were detected. According to the response to hormone therapy, NS children were divided into the hormone resistant nephrotic syndrome (SRNS) group (31 cases) and the hormone sensitive nephrotic syndrome (SSNS) group (126 cases). Factors influencing the resistance of NS children to hormone therapy were analyzed by Logistic regression, and the differential diagnositic value of ET-1 and SDC-1 to SRNS and SSNS were analyzed by receiver operating characteristic curve (ROC). Results The levels of body weight, BUN, Scr, 24 h urine protein, ET-1 and SDC-1 were increased successively in the control group, the SSNS group and the SRNS group, while the serum albumin was decreased successively (P<0.05). Serum ET-1 and SDC-1 levels of all the subjects were positively correlated with BUN, Scr and 24 h urinary protein, and negatively correlated with serum albumin level (P<0.05). Multivariate Logistic regression analysis showed that the increased ET-1 and SDC-1 were risk factors for the resistance of NS children to hormone therapy (P<0.05). ROC analysis results showed that the area under the curve for the identification of SRNS and SSNS combined with ET-1 and SDC-1 was 0.943, which was higher than ET-1 and SDC-1 (Z=3.959, 4.364, P<0.05). Conclusion Serum ET-1 and SDC-1 levels are significantly increased in children with SRNS, which are related to renal function injury and resistance to steroid therapy, and can be used as potential indicators for the differential diagnosis of SRNS.

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    The clinical significance of serum levels of GDF-15 and hs-CRP in patients with coronary slow flow
    YAO Kunmei, XIA Zhonghua, HUANG Xingjie, HUANG Min, LIU Qiong, ZHOU Hanli
    2023, 51 (4):  418-421.  doi: 10.11958/20221391
    Abstract ( 360 )   HTML ( 4 )   PDF (719KB) ( 922 )  

    Objective To investigate changes and clinical significance of serum growth differentiation factor-15 (GDF-15) and hypersensitive C-reactive protein (hs-CRP) in patients with coronary slow flow (CSF). Methods A total of 132 subjects were selected and divided into the CSF group (n=66) and the control group (n=66) according to results of coronary arteriography (CAG). The serum levels of GDF-15 and hs-CRP were compared between the two groups. The correlation between serum levels of GDF-15 and hs-CRP and coronary blood flow mTFC was analyzed by Spearman correlation analysis. Binary Logistic regression was used to analyze the influencing factors of CSF. Results Compared with the control group, serum levels of GDF-15 [969.01(620.39, 1 191.01) ng/L vs. 552.73(197.06, 776.09) ng/L], hs-CRP [5.36(2.57, 6.77) mg/L vs. 0.96(0.49, 3.47) mg/L] and mTFC [27.38(23.17, 33.20) vs. 16.52(14.13, 19.12)] were significantly higher in the CSF group (P<0.05). The serum levels of GDF-15 and hs-CRP were positively correlated with mTFC in the CSF group (rs = 0.417 and 0.336, P<0.01). Binary Logistic regression analysis showed that increased GDF-15 (OR=1.004, 95%CI: 1.002-1.006, P<0.001) and hs-CRP (OR=1.604, 95%CI: 1.264-2.035, P<0.001) were independent risk factors for CSF. Conclusion The serum levels of GDF-15 and hs-CRP are elevated in CSF patients. There is a positive linear correlation between GDF-15 and hs-CRP with mTFC, which can reflect the severity of CSF.

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    Prediction of risk factors for hypothyroidism in NSTE-ACS patients exposed to iodine contrast media
    HAN Chuyi, CONG Hongliang, WANG Le, ZHANG Jingxia
    2023, 51 (4):  422-426.  doi: 10.11958/20221362
    Abstract ( 331 )   HTML ( 1 )   PDF (810KB) ( 942 )  

    Objective To explore the effect of iodine contrast media (ICM) on thyroid function and the risk factors of hypothyroidism in patients with non-ST segment elevation acute coronary syndrome (NSTE-ACS) after percutaneous coronary intervention (PCI). Methods According to the baseline thyroid stimulating hormone (TSH) level, 627 patients were divided into the hypothyroidism group (group H, n=62) and the euthyroid group (group ET, n=565). Changes of thyroid function were monitored at 1 day, 1 week, 1 month and 6 months after PCI, and the risk factors of hypothyroidism before and after PCI were analyzed by multivariate analysis. Results The levels of TSH increased significantly on the 1 day and 1 week after PCI in both groups (P<0.05), and group ET was still higher than its baseline value at 6 months (P<0.05). At 6 months, 43 of 62 cases in the group H were still in the state of hypothyroidism, but the event of overt hypothyroidism did not increase. In the group ET, there were 133 cases of ICM-induced hypothyroidism (23.9%), 34 cases of hypothyroidism and 4 cases of iodine-induced hyperthyroidism at 6 months. The independent predictors for hypothyroidism after ICM were not entirely consistent with the independent predictors for baseline hypothyroidism. The dosage of ICM was a reliable predictor for long-term hypothyroidism after PCI (AUC=0.674), and the best security threshold was 190 mL. Conclusion Patients with NSTE-ACS have a lower risk of developing overt hypothyroidism after ICM exposure. Taking 6 months after ICM exposure as a time window for study evaluation may mask some events of prolonged ICM-induced thyroid insufficiency.

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    Expression of serum levels of CXCL9 and SFRP1 in patients with AECOPD complicated with pulmonary embolism and their influence on prognosis
    SU Zhenlei, YANG Wanchun, HAN Tianlong
    2023, 51 (4):  427-431.  doi: 10.11958/20221267
    Abstract ( 340 )   HTML ( 4 )   PDF (765KB) ( 944 )  

    Objective To explore expression levels of serum C-X-C chemokine ligand 9 (CXCL9) and secretory frizzled-associated protein 1 (SFRP1) in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) complicated with pulmonary embolism (PE) and their prognostic value. Methods Fifty-six AECOPD patients with PE were collected as the case group, and 80 patients with AECOPD in the same period were selected as the control group. The serum CXCL9 and SFRP1 levels were detected by enzyme-linked immunosorbent assay. Logistic regression model was used to analyze the influencing factors of PE in AECOPD patients, and the receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of serum CXCL9 and SFRP1 on PE in AECOPD patients. COX proportional risk regression model was used to analyze the influencing factors of serum CXCL9 and SFRP1 on poor prognosis. Results The course of AECOPD was longer in the case group than that in the control group, and the serum CXCL9 and SFRP1 levels were higher than those in the control group (P<0.05). Long AECOPD course (OR=1.420,95%CI:1.246-1.620), higher CXCL9 (OR=1.835,95%CI:1.502-2.241) and SFRP1 (OR=1.608,95%CI:1.351-1.914) were independent risk factors for PE in AECOPD patients. Serum levels of CXCL9 and SFRP1 can predict the risk of PE in AECOPD patients, and the predictive value of combination of two indicators was more better. Serum CXCL9 and SFRP1 levels were positively associated with PESI grade (rs=0.752, 0.716; P<0.05). Higher serum levels of CXCL9 (HR=1.374, 95%CI:1.184-1.595) and SFRP1 (HR=1.186,95%CI:1.329-1.542) were independent risk factors for death in AECOPD patients with PE. Conclusion The abnormal increased serum levels of CXCL9 and SFRP1 are risk factors for PE in AECOPD patients, which is closely related to the severity and prognosis of PE.

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    Clinical observation of the modified Broström procedure augmented with suture-tape repair of anterior talofibular ligament injury using mini-open incision
    SUN Ruobin, SHANG Lin, XU Shaogang
    2023, 51 (4):  432-435.  doi: 10.11958/20221841
    Abstract ( 554 )   HTML ( 1 )   PDF (1094KB) ( 921 )  

    Objective To explore the surgical treatment of the modified Broström procedure augmented with suture-tape repair of anterior talofibular ligament using mini-open incision. Methods A tatal of 17 patients with chronic lateral ankle instability underwent the modified Broström procedure augmented with suture-tape reconstruction of anterior talofibular ligament using mini-open incision. The patients included 11 males and 6 females, aged 28-56 years with a mean of (40.20±13.75) years. Body mass index (BMI) was 30.12-35.10 kg/m2 with a mean of (32.27±1.83) kg/m2. The FAOS and FAAM were used to evaluate the postoperative function of ankle. Results All patients had operation performed smoothly, without serious complications, and got incisions healed well. All 17 patients were followed up for 12-24 months. Compared with those preoperative, the FAOS (from 57.10±11.32 to 91.35±1.27, P<0.05) and FAAM (from 55.76±10.12 to 91.10±0.69, P<0.05) scores significantly improved at the latest follow-up and no complications, such as nerve, blood vessel and tendon injury. Two patients experienced tightness in the lateral ankle during early walking, and the symptoms were resolved after 8 weeks. Conclusion For the chronic lateral ankle instability, the modified Broström procedure augmented with suture-tape repair of anterior talofibular ligament using mini-open incision effectively reduces symptoms and get early rehabilitation, with definite effect.

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    Applied Research
    Clinical analysis and prediction of intrauterine fetal death in late trimester of pregnancy
    ZHAO Caiyun, HUA Shaofang
    2023, 51 (4):  436-440.  doi: 10.11958/20221476
    Abstract ( 388 )   HTML ( 3 )   PDF (803KB) ( 921 )  

    Objective To investigate the risk and perinatal outcomes of intrauterine fetal death (IUFD) in late trimester of pregnancy and to construct and verify the nomogram risk prediction model. Methods Sixty-nine cases of IUFD in late trimester of pregnancy were selected as the case group, and 69 cases with normal delivery during the same period were selected as the control group. Clinical data such as general condition, maternal factors, fetal factors, fetal appendage factors and perinatal outcomes were compared between the two groups. Logistic regression was used to analyze risk factors of fetal death in the third trimester of pregnancy. The nomogram model for risk prediction was established, and the Bootstrap method was used for internal verification. The C-Statistic was calculated, and the goodness of fit of the model was evaluated by Hosmer-Lemeshow test. Results The age of the case group was higher than that of the control group, and the frequency of childbirth examination was less than that of the control group (P<0.05). The proportion of non-higher education background, non-urban living, unemployed, elderly (≥35 years old), multiparous women, conscious abnormal fetal movement, preeclampsia, twin pregnancy, intrauterine infection, abnormal ultrasound of fetal system, chromosomal abnormality, small for gestational age, abnormality of umbilical cord, amniotic fluid volume anomaly and the proportion of premature and vaginal birth were significantly higher in the case group than those of the control group (P<0.05). Logistic regression analysis showed that conscious abnormal fetal movement, preeclampsia, twin pregnancy, abnormal ultrasound of fetal system or chromosome abnormality and umbilical cord or amniotic fluid abnormality were independent risk factors for fetal death in late pregnancy, and the higher frequency of childbirth examination was its protective factor (P<0.05). Based on results of multivariate Logistic regression analysis, age was included in the prediction model combined with the actual clinical situation, and the nomogram risk prediction model was established. Results of Bootstrap internal verification and Hosmer-Lemeshow test showed that the discrimination (C-Statistic=0.937) and calibration of the model were good (χ2=5.364, P=0.643). Conclusion The nomogram model can effectively assess the risk of fetal death in late trimester of pregnancy and has good clinical application value.

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    Review
    Research progress on the relationship between high fat diet and cognitive impairment
    LI Xiaoxiao, BAI Yanjie, WANG Yan, ZHANG Yongchuang, CHEN Limin, CHEN Shuying
    2023, 51 (4):  441-444.  doi: 10.11958/20221298
    Abstract ( 372 )   HTML ( 4 )   PDF (718KB) ( 949 )  

    Cognitive impairment refers to one or more impairments in feeling, learning and memory, speech, attention and execution, which seriously affects the quality of life of patients. Studies have shown that high fat diet (HFD) can cause neuronal damage and cerebrovascular dysfunction, thus causing cognitive impairment. This article reviews the relevant mechanisms and treatment progress of HFD induced cognitive impairment, with a view to providing new ideas for effective prevention and treatment of cognitive impairment.

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    Research progress in clinical application of choroid capillary flow deficits
    BI Meng, FAN Fang, JIA Zhiyang
    2023, 51 (4):  445-448.  doi: 10.11958/20221642
    Abstract ( 274 )   HTML ( 3 )   PDF (733KB) ( 894 )  

    With the development of optical coherence tomography (optical coherence tomography angiography, OCTA), quantitative study of choriocapillaris (choriocapillaris, CC) has become a novel evaluation method. The occurrence and development of eye diseases, such as retina and choroid are closely related to the morphology and function of choroidal vessels, especially the change of choriocapillaris. The CC flow deficits parameters, such as density, area and number can be obtained after processing and quantitative analysising images of CC layer from OCTA, assess changes in choroid blood capillary perfusion, explore the pathogenesis of related diseases and provide certain help for disease diagnosis and intervention therapy. This paper reviews the clinical application and research progress of choroid capillary flow deficits.

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