Liver fibrosis is a chronic liver injury resulting from factors like viral hepatitis, autoimmune hepatitis, non-alcoholic steatohepatitis, fatty liver disease, and cholestatic liver disease. Liver transplantation is currently the gold standard for treating severe liver diseases. However, it is limited by a shortage of donor organs and the necessity for lifelong immunosuppressive therapy. Mesenchymal stem cells (MSCs) can differentiate into various liver cells and enhance liver function when transplanted into patients due to their differentiation and proliferation capabilities. Therefore, it can be used as an alternative therapy for treating liver diseases, especially for liver cirrhosis, liver failure, and liver transplant complications. However, due to the potential tumorigenic effects of MSCs, researchers are exploring a new approach to treating liver fibrosis using extracellular vesicles (exosomes) secreted by stem cells. Many studies show that exosomes released by stem cells can promote liver injury repair through various pathways, contributing to the treatment of liver fibrosis. In this review, we focus on the molecular mechanisms by which stem cell exosomes affect liver fibrosis through different pathways and their potential therapeutic targets. Additionally, we discuss the advantages of exosome therapy over stem cell therapy and the possible future directions of exosome research, including the prospects for clinical applications and the challenges to be overcome.

Liver fibrosis is a chronic liver injury resulting from factors like viral hepatitis, autoimmune hepatitis, non-alcoholic steatohepatitis, fatty liver disease, and cholestatic liver disease. Liver transplantation is currently the gold standard for treating severe liver diseases. However, it is limited by a shortage of donor organs and the necessity for lifelong immunosuppressive therapy. Mesenchymal stem cells (MSCs) can differentiate into various liver cells and enhance liver function when transplanted into patients due to their differentiation and proliferation capabilities. Therefore, it can be used as an alternative therapy for treating liver diseases, especially for liver cirrhosis, liver failure, and liver transplant complications. However, due to the potential tumorigenic effects of MSCs, researchers are exploring a new approach to treating liver fibrosis using extracellular vesicles (exosomes) secreted by stem cells. Many studies show that exosomes released by stem cells can promote liver injury repair through various pathways, contributing to the treatment of liver fibrosis. In this review, we focus on the molecular mechanisms by which stem cell exosomes affect liver fibrosis through different pathways and their potential therapeutic targets. Additionally, we discuss the advantages of exosome therapy over stem cell therapy and the possible future directions of exosome research, including the prospects for clinical applications and the challenges to be overcome.

Liver fibrosis is a chronic liver injury resulting from factors like viral hepatitis, autoimmune hepatitis, non-alcoholic steatohepatitis, fatty liver disease, and cholestatic liver disease. Liver transplantation is currently the gold standard for treating severe liver diseases. However, it is limited by a shortage of donor organs and the necessity for lifelong immunosuppressive therapy. Mesenchymal stem cells (MSCs) can differentiate into various liver cells and enhance liver function when transplanted into patients due to their differentiation and proliferation capabilities. Therefore, it can be used as an alternative therapy for treating liver diseases, especially for liver cirrhosis, liver failure, and liver transplant complications. However, due to the potential tumorigenic effects of MSCs, researchers are exploring a new approach to treating liver fibrosis using extracellular vesicles (exosomes) secreted by stem cells. Many studies show that exosomes released by stem cells can promote liver injury repair through various pathways, contributing to the treatment of liver fibrosis. In this review, we focus on the molecular mechanisms by which stem cell exosomes affect liver fibrosis through different pathways and their potential therapeutic targets. Additionally, we discuss the advantages of exosome therapy over stem cell therapy and the possible future directions of exosome research, including the prospects for clinical applications and the challenges to be overcome.

OBJECTIVES: The purinergic receptor P2X2 (P2RX2) encodes an ATP-gated ion channel permeable to Na⁺, K⁺, and especially Ca²⁺. Loss-of-function mutations in P2RX2 are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a p2rx2 knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of p2rx2 deficiency on the auditory system, providing a basis for understanding P2RX2-related hearing loss and developing gene therapy strategies. METHODS: Two CRISPR targets (sgRNA1 and sgRNA2) spaced 47 bp apart were designed within the zebrafish p2rx2 gene. Synthesized sgRNAs and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group. RESULTS: A stable p2rx2 knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all P<0.001), indicating reduced hearing sensitivity. CONCLUSIONS We successfully generated a p2rx2 loss-of-function zebrafish model using CRISPR/Cas9 technology. p2rx2 deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotype's restriction to the otolith region suggests tissue-specific roles of p2rx2 in sensory organs. This model provides a valuable tool for elucidating the molecular mechanisms of P2RX2-related hearing loss and for screening otoprotective drugs and developing gene therapies.
Animals ; Zebrafish/genetics* ; Receptors, Purinergic P2X2/deficiency* ; CRISPR-Cas Systems/genetics* ; Gene Knockout Techniques ; Phenotype ; Zebrafish Proteins/genetics* ; Disease Models, Animal

Objective:To investigate expression and clinical significance of T cell activation related membrane molecules in infertility patients with polycystic ovary syndrome(PCOS).Methods:A total of 80 PCOS infertility patients who visited Second Affiliated Hospital of Hainan Medical College from January 2020 to June 2021 were selected as observation group,and 80 cases of women who underwent physical examination at same time were selected as control group.Expressions of membrane molecules related to T cell acti-vation in peripheral blood of patients between two groups were compared.Patients with PCOS infertility were divided into pregnant group[66(82.5％)]and non pregnant group[14(17.5％)]according to different prognosis.General clinical data,peripheral blood inflammatory factors and T cell activation related membrane molecules expressions were compared between two groups,focusing on correlation between T cell activation related membrane molecules expressions and prognosis of PCOS infertility patients,ROC curve and decision-making curve of T cell activation related membrane molecule expressions were drawn to predict prognosis of PCOS infer-tility patients,whose prediction efficiency and net profitability were further analyzed.Pearson correlation analysis was used to explore correlation between T cell activation related membrane molecules expressions and clinical characteristics of PCOS infertility patients.Results:CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood of patients in observation group were significantly higher than control group(P<0.05).Total testosterone(T),TNF-α,IFN-γ,CD4+CD45+T and CD4+CD63+T cells expressions of patients in non pregnancy group were significantly higher than pregnancy group(P<0.05).Multivariate Logistic regression analysis showed that CD4+CD45+T and CD4+CD63+T cells were independent predictors of prognosis of PCOS infertility patients(P<0.05),ROC analysis showed that AUC of CD4+CD45+T cells predicting prognosis of PCOS infertility patients was 0.756(0.721～0.826),and the best diag-nostic cut-off point was 48.1％,AUC of CD4+CD63+T cells was 0.746(0.711～0.834),and the best cut-off point for diagnosis was 31.2％,while AUC of CD4+CD45+T and CD4+CD63+T cells combination was 0.948(0.897～0.986).Decision curve analysis showed that combined prediction of CD4+CD45+T and CD4+CD63+T cells was higher than a single indicator.Correlation analysis showed that CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood of patients with PCOS infertility was related to T,TNF-α,insulin resistance index(HOMA-IR),IFN-γ,IL-2 and IL-10(P<0.05).Conclusion:The higher the CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood,the worse the prognosis of patients with PCOS infertility.

OBJECTIVE To systematically evaluate the risk factors for spontaneous bacterial peritonitis(SBP)in the cirrhosis patients with ascites so as to provide bases for reducing the incidence of SBP.METHODS The literatures regarding to cirrhosis ascites complicated with SBP were retrieved from the databases such as PubMed,Embase,Cochrane Library,CBM,CNKI,Wanfang and VIP.The time frame of retrieval was limited from the establish-ment of the databases to Jan.10,2024.The quality of the literatures was evaluated by Newcastle-Ottawa Scale(NOS).The data were analyzed by using STATA 16.0.RESULTS A total of 43 pieces of literatures were included in the study,involving 9877 research subjects,3384 of whom were complicated with SBP.The result of meta a-nalysis showed that the risk factors(P＜0.05)for SBP in the cirrhosis patients with ascites included the general factor(history of SBP),the disease factors(gastrointestinal hemorrhage,diarrhea,diabetes mellitus),drug fac-tors(use of proton pump inhibitors),physiological indexes[low blood sodium,low serum albumin,high serum bilirubin,rise of C-reactive protein(CRP)],liver function scoring indexes[Child-turcotte-pugh(CTP)grade C,model of end-stage liver disease(MELD)],and ascites indexes[low ascites albumin,rise of ascites white blood cells and rise of tumor necrosis factor-α(TNF-α)].CONCLUSIONS SBP is one of the most severe complications in the cirrhosis patients with ascites.Multiple risk factors may have remarkable influence on the occurrence and de-velopment of SBP.It is necessary to take targeted interventions to the controllable risk factors during the clinical practice so as to effectively reduce the risk of SBP in the cirrhosis patients with ascites.

Objective:To observe the changes of central visual acuity and extracentral visual acuity in eyes with non-arteritic central retinal artery occlusion (NA-CRAO).Methods:A retrospective clinical study. From January 1, 2017 to December 31, 2024, 140 patients (140 eyes) diagnosed with NA-CRAO through ophthalmic examination at Department of Ophthalmology of First People's Hospital of Xianyang City were included in the study. All affected eyes underwent best corrected visual acuity (BCVA), visual field, intraocular pressure, fundus color photography, optical coherence tomography (OCT), and fluorescein angiography (FFA) examinations. After a clear diagnosis, conservative treatment such as reducing intraocular pressure, relieving spasms, and dilating blood vessels should be given immediately. Simultaneously, intravenous and/or arterial thrombolysis therapy should be administered based on the patient's overall condition. Under the same treatment conditions as other treatments, 33 eyes were treated with hyperbaric oxygen therapy within 24 hours after seeking medical attention. The changes in central visual acuity (BCVA) and peripheral visual acuity of the affected eye one month after treatment were observed. BCVA improvement of ≥ 1 line was defined as the increase of no light sensitivity to light sensitivity or above, and the increase of light sensitivity to 0.01 or above. The visual acuity outside the center was determined by the 0 ° axis in front of the eyeball at eye level, and was 10 ° outside visual acuity on the temporal side. Multivariate analysis using logistic regression analysis.Results:Among the 140 cases (140 eyes), there were 84 males (84 eyes) and 56 females (56 eyes). The mean age was (63.89±10.78) years. The duration of illness from the onset of symptoms to the time of diagnosis was 48 (2-720) hours. 6, 1, 14, 47, 41, 16, and 15 eyes were diagnosed with BCVA without light perception, uncertain light perception, manual/anterior, digital/anterior, 0.01-0.10, and ≥ 0.10, respectively. FFA examination revealed delayed arm retinal circulation time and filling of the retinal artery trunk to the peak, with changes in the "arterial front" observed in 126 eyes. OCT examination showed extensive edema and unclear structure in the inner layer of the retina in all patients. Out of 140 eyes, 122 were treated with intravenous thrombolysis and 4 with arterial thrombolysis; 14 eyes did not receive thrombolytic therapy. After treatment, 38 eyes (27.1 %) showed an improvement of BCVA ≥ 1; 67 eyes (47.9%) did not show an improvement in BCVA, and the affected eye had a BCVA of approximately 0.6 without light perception; 17 eyes (12.1 %) showed improvement in peripheral vision, and the peripheral vision of the affected eyes ranged from 0.01 to 0.1, all of whom were patients undergoing intravenous thrombolysis, and prior to treatment, this group of patients had complete blindness in the coarse side visual field of the Amsler grid, and their out of center visual acuity could not be measured. Among the 33 eyes treated with hyperbaric oxygen therapy, 24 eyes (72.7%) showed an increase in BCVA after treatment; 9 eyes did not improve, among which 4 eyes (12.1%) showed improvement in out of center visual acuity. Among the 107 eyes that did not receive hyperbaric oxygen therapy, 49 eyes (45.8%) showed an increase in BCVA after treatment. There was no improvement in 58 eyes (54.2%), among which 13 eyes (12.1%) showed an improvement in out of center visual acuity. The results of logistic regression analysis showed that intravenous thrombolysis and hyperbaric oxygen therapy were independent predictive factors for the improvement of central and extra central visual acuity ( P<0.05). Conclusions:Hyperbaric oxygen therapy within 24 hours of seeking medical attention for patients with NA-CRAO disease course ≤ 1 month has a significant effect on the recovery of central and extra central vision. Intravenous thrombolysis and hyperbaric oxygen therapy are independent predictive factors for the improvement of central and extra central vision.

OBJECTIVE To systematically evaluate the risk factors for spontaneous bacterial peritonitis(SBP)in the cirrhosis patients with ascites so as to provide bases for reducing the incidence of SBP.METHODS The literatures regarding to cirrhosis ascites complicated with SBP were retrieved from the databases such as PubMed,Embase,Cochrane Library,CBM,CNKI,Wanfang and VIP.The time frame of retrieval was limited from the establish-ment of the databases to Jan.10,2024.The quality of the literatures was evaluated by Newcastle-Ottawa Scale(NOS).The data were analyzed by using STATA 16.0.RESULTS A total of 43 pieces of literatures were included in the study,involving 9877 research subjects,3384 of whom were complicated with SBP.The result of meta a-nalysis showed that the risk factors(P＜0.05)for SBP in the cirrhosis patients with ascites included the general factor(history of SBP),the disease factors(gastrointestinal hemorrhage,diarrhea,diabetes mellitus),drug fac-tors(use of proton pump inhibitors),physiological indexes[low blood sodium,low serum albumin,high serum bilirubin,rise of C-reactive protein(CRP)],liver function scoring indexes[Child-turcotte-pugh(CTP)grade C,model of end-stage liver disease(MELD)],and ascites indexes[low ascites albumin,rise of ascites white blood cells and rise of tumor necrosis factor-α(TNF-α)].CONCLUSIONS SBP is one of the most severe complications in the cirrhosis patients with ascites.Multiple risk factors may have remarkable influence on the occurrence and de-velopment of SBP.It is necessary to take targeted interventions to the controllable risk factors during the clinical practice so as to effectively reduce the risk of SBP in the cirrhosis patients with ascites.