To investigate the effects of plasma exosomes from sheep on animal hair growth,exo-somes were extracted from two sheep breeds with significant differences in wool fineness(Xinji Fine-wool sheep and Small-tailed Han sheep)using biphasic precipitation.The exosomes were characterized by transmission electron microscopy(TEM)and nanoparticle tracking analysis(NTA).A murine alopecia areata model was established,followed by subcutaneous injection of ex-osomes for 7 consecutive days.Dorsal skin samples were collected on day 10 post-injection for his-tological analysis via hematoxylin-eosin(HE)staining.miRNA and mRNA libraries were constructed through qPCR,while TargetScan and g:Profiler were employed for target gene predic-tion and KEGG enrichment analysis,respectively.Protein-protein interaction(PPI)networks were analyzed using STRING to identify key miRNA-mRNA targets,which were preliminarily validated by qPCR.Results demonstrated that on day 5 post-injection,the Small-tailed Han sheep plasma ex-osome group(SPE)exhibited more melanin deposition spots compared to the Xinji Fine-wool sheep plasma exosome group(XPE),with XPE showing greater melanin spots than the control group(NC).Histological analysis revealed that XPE group displayed significantly fewer hair folli-cles compared to SPE and NC groups(P＜0.01),while SPE group exhibited significantly larger hair follicle diameters than both XPE and NC groups(P＜0.01).Bioinformatics analysis predicted 357,711,477,346,and 3 178 target genes for miR-150,miR-133b,miR-31-5p,miR-433-3p,and miR-218,respectively,with 508 genes being co-regulated by two or more miRNAs.PPI analysis i-dentified 217 genes involved in positive regulation of cellular processes,109 in developmental regu-lation,133 in cellular development,132 in cell differentiation,and 69 in regulation of cell differenti-ation.Target genes were found to regulate hair follicle growth through Wnt signaling via Frizzled receptor binding and downstream pathways involving calmodulin(CaM)and cyclic-AMP response binding protein(CREB).qPCR validation showed significantly higher relative expression levels of miR-218,miR-150,miR-31-5p,miR-133b,and miR-433-3p in SPE and XPE groups compared to NC,with SPE exhibiting greater expression than XPE.Additionally,significant differences in FZD4,WNT4,CREB1,and FZD3 expression were observed between treatment groups and NC.These findings demonstrate that plasma exosomes from Xinji Fine-wool and Small-tailed Han sheep modulate hair growth by altering miRNA-mRNA expression profiles in skin tissues.

Objective:To analyze the efficacy of endovascular treatment for venous outflow tract obstruction after liver transplantation.Methods:A retrospective analysis was conducted on the data of 7 patients with venous outflow tract obstruction after liver transplantation admitted to Beijing Friendship Hospital, Capital Medical University from November 2020 to December 2024. Among them, there were 5 males and 2 females, with the age of (22.3±8.1) years. The primary diseases included 2 cases of Budd-Chiari syndrome, 1 case of hepatic veno-occlusive disease, 1 case of portal veno-hepatic sinus vascular disease, 1 case of ornithine carbamoyltransferase deficiency, 1 case of primary biliary cirrhosis, and 1 case of autoimmune cirrhosis. Analyze the patient's clinical manifestations, obstruction of venous outflow tract, hemoglobin levels within one week before and one week after the operation, endovascular treatment conditions, and intraoperative complications such as abdominal hemorrhage and vascular injury. Ultrasound was used to measure the depth of ascites and pleural effusion. All patients were followed up immediately after the operation through phone calls or follow-up visits. The clinical symptoms, abdominal vascular ultrasound, enhanced CT and survival status of the patients were followed up.Results:All 7 patients were diagnosed with venous outflow tract obstruction by intraoperative angiography, including 1 cases of inferior vena cava obstruction, 2 cases of hepatic vein obstruction, and 4 cases of vena cava combined with hepatic vein obstruction. A total of 12 endovascular treatments were performed on 7 patients. Among them, 4 patients received balloon dilation and angioplasty once, 1 patient received balloon dilation and angioplasty twice, 1 patient underwent hepatic vein stent implantation after 2 hepatic vein balloon dilation and angioplasty, and 1 patient underwent intrahepatic portosystemic shunt via jugular vein after 2 hepatic vein balloon dilation and angioplasty. The abdominal distensionof the patients were all relieved after the operation, the ascites and pleural effusion decreased, and the edema symptoms of the lower extremities disappeared. There were no intraoperative complications. The preoperative hemoglobin of 7 patients was (113.4±34.0) g/L, and the postoperative hemoglobin was (126.6±34.8) g/L, which increased significantly compared with that before the operation, and the difference was statistically significant ( t=-0.71, P=0.038). Seven patients were followed up for 6 to 24 months, with a median of 12 months. None of them had obvious symptoms including abdominal distension. Abdominal ultrasound and CT indicated that the blood flow of the transplanted liver was unobstructed, and no patient died. Conclusion:Venous outflow tract obstruction after liver transplantation can cause severe symptoms. Endovascular treatment is an effective treatment for venous outflow tract obstruction after liver transplantation.

Based on network pharmacology,molecular docking technology and experimental valida-tion to explore the therapeutic efficacy and mechanism of action of baicalin(BC)on kidney injury caused by Klebsiella pneumoniae(KP)infection.The inhibitory activity of BC against KP was de-termined by in vitro experiments;a mouse kidney injury model was established,and the therapeu-tic effect was preliminarily verified by ophthalmoscopy and pathological histology;three pro-in-flammatory factors,namely,TNF-α,IL-10,and IL-1β,were detected by ELISA;and the cyber-pharmacology technology was utilized by PubChem,TCMSP,STRING,Cytoscape,AutoDocks and other databases and software to construct the PPI network as well as to perform GO function and KEGG enrichment analyses;and molecular docking technology was used to assess the binding ac-tivity of the drugs to the core targets and to speculate on the signaling pathways of the drug action.The results showed that BC had a better inhibitory effect on KP in the in vitro experiments;path-ological histology showed a significant therapeutic effect of BC;compared with the infected group,the content of pro-inflammatory factors TNF-α,IL-10,and IL-1β in the baicalin treatment group were significantly decreased(P≤0.05).Twenty-four core targets and 11 pathways of action were screened by network pharmacology,and BC docked stably with the acquired core targets TP53,PTGS2,MAPK1,MAPK8,TNF,BCL2,and IGF1 molecules,and it was speculated that BC might exert its antibacterial and anti-inflammatory effects through the signaling pathways of PI3K-Akt,MAPK,HIF-1,and NF-kappa B,etc.This study lays the foundation for further research on the mechanism of action of baicalin on renal injury.

To investigate the pathogenicity of Sheep-origin Proteus mirabilis and the immunity effect of its inactivated vaccine.In this study,autopsy were performed on sick and dead sheep with severe diarrhea,collected liver,spleen and other diseased organs.And identified the isolated strains by morphology,Gram staining microscopy,biochemical test,molecular biology and 16S rRNA se-quencing;Drug resistance and pathogenicity were studied by drug sensitivity test,pathogenicity test in mouse and pathological histological observation;The isolated strains were inactivated by formaldehyde and mixed with alum adjuvant containing aluminum hydroxide to prepare the inacti-vated vaccine;mice were immunized after safety testing to test antibodies,cytokines and evaluate the immunological effects.Results showed that the isolate was Gram-negative short bacillus with blunt rounded ends after purification staining.Combined with biochemical tests,PCR amplified fragments and 16S rRNA sequencing results showed that the strain was Proteus mirabilis;The re-sults of drug sensitivity test showed that the isolate was highly sensitive to four antibiotics,including amikacin and piperacillin(S),the rest of the strain was moderately or poorly sensitive,or even resistant;The isolate was highly sensitive to the four antibiotics,such as amikacin and pip-eracillin(S).The lethal concentration of the isolated strain was 1.6 × 105 CFU/mL and it could cause different degrees of congestion and bruising and inflammatory cell infiltration in the liver,spleen and other major organs;The formaldehyde inactivated vaccine did not cause adverse reac-tions in mice,and the serum levels of antibodies to IgG,IgG1 and IgG2a,and the levels of antibod-ies to IL-4 and IFN-γ were higher than that of the PBS group,and the vaccine immunoprotection rate was 90％in the attacking bacterial test.In the present experiment,Proteus mirabilis was suc-cessfully isolated from the organs of sick and dead sheep.The inactivated vaccine prepared by form-aldehyde inactivation method can mediate humoral immunity to play a preventive role against the infection of S.chimaera,which can provide a new method for the prevention of bacterial diseases in sheep breeding.

To investigate the pathogenicity of Sheep-origin Proteus mirabilis and the immunity effect of its inactivated vaccine.In this study,autopsy were performed on sick and dead sheep with severe diarrhea,collected liver,spleen and other diseased organs.And identified the isolated strains by morphology,Gram staining microscopy,biochemical test,molecular biology and 16S rRNA se-quencing;Drug resistance and pathogenicity were studied by drug sensitivity test,pathogenicity test in mouse and pathological histological observation;The isolated strains were inactivated by formaldehyde and mixed with alum adjuvant containing aluminum hydroxide to prepare the inacti-vated vaccine;mice were immunized after safety testing to test antibodies,cytokines and evaluate the immunological effects.Results showed that the isolate was Gram-negative short bacillus with blunt rounded ends after purification staining.Combined with biochemical tests,PCR amplified fragments and 16S rRNA sequencing results showed that the strain was Proteus mirabilis;The re-sults of drug sensitivity test showed that the isolate was highly sensitive to four antibiotics,including amikacin and piperacillin(S),the rest of the strain was moderately or poorly sensitive,or even resistant;The isolate was highly sensitive to the four antibiotics,such as amikacin and pip-eracillin(S).The lethal concentration of the isolated strain was 1.6 × 105 CFU/mL and it could cause different degrees of congestion and bruising and inflammatory cell infiltration in the liver,spleen and other major organs;The formaldehyde inactivated vaccine did not cause adverse reac-tions in mice,and the serum levels of antibodies to IgG,IgG1 and IgG2a,and the levels of antibod-ies to IL-4 and IFN-γ were higher than that of the PBS group,and the vaccine immunoprotection rate was 90％in the attacking bacterial test.In the present experiment,Proteus mirabilis was suc-cessfully isolated from the organs of sick and dead sheep.The inactivated vaccine prepared by form-aldehyde inactivation method can mediate humoral immunity to play a preventive role against the infection of S.chimaera,which can provide a new method for the prevention of bacterial diseases in sheep breeding.

To investigate the effects of plasma exosomes from sheep on animal hair growth,exo-somes were extracted from two sheep breeds with significant differences in wool fineness(Xinji Fine-wool sheep and Small-tailed Han sheep)using biphasic precipitation.The exosomes were characterized by transmission electron microscopy(TEM)and nanoparticle tracking analysis(NTA).A murine alopecia areata model was established,followed by subcutaneous injection of ex-osomes for 7 consecutive days.Dorsal skin samples were collected on day 10 post-injection for his-tological analysis via hematoxylin-eosin(HE)staining.miRNA and mRNA libraries were constructed through qPCR,while TargetScan and g:Profiler were employed for target gene predic-tion and KEGG enrichment analysis,respectively.Protein-protein interaction(PPI)networks were analyzed using STRING to identify key miRNA-mRNA targets,which were preliminarily validated by qPCR.Results demonstrated that on day 5 post-injection,the Small-tailed Han sheep plasma ex-osome group(SPE)exhibited more melanin deposition spots compared to the Xinji Fine-wool sheep plasma exosome group(XPE),with XPE showing greater melanin spots than the control group(NC).Histological analysis revealed that XPE group displayed significantly fewer hair folli-cles compared to SPE and NC groups(P＜0.01),while SPE group exhibited significantly larger hair follicle diameters than both XPE and NC groups(P＜0.01).Bioinformatics analysis predicted 357,711,477,346,and 3 178 target genes for miR-150,miR-133b,miR-31-5p,miR-433-3p,and miR-218,respectively,with 508 genes being co-regulated by two or more miRNAs.PPI analysis i-dentified 217 genes involved in positive regulation of cellular processes,109 in developmental regu-lation,133 in cellular development,132 in cell differentiation,and 69 in regulation of cell differenti-ation.Target genes were found to regulate hair follicle growth through Wnt signaling via Frizzled receptor binding and downstream pathways involving calmodulin(CaM)and cyclic-AMP response binding protein(CREB).qPCR validation showed significantly higher relative expression levels of miR-218,miR-150,miR-31-5p,miR-133b,and miR-433-3p in SPE and XPE groups compared to NC,with SPE exhibiting greater expression than XPE.Additionally,significant differences in FZD4,WNT4,CREB1,and FZD3 expression were observed between treatment groups and NC.These findings demonstrate that plasma exosomes from Xinji Fine-wool and Small-tailed Han sheep modulate hair growth by altering miRNA-mRNA expression profiles in skin tissues.

Based on network pharmacology,molecular docking technology and experimental valida-tion to explore the therapeutic efficacy and mechanism of action of baicalin(BC)on kidney injury caused by Klebsiella pneumoniae(KP)infection.The inhibitory activity of BC against KP was de-termined by in vitro experiments;a mouse kidney injury model was established,and the therapeu-tic effect was preliminarily verified by ophthalmoscopy and pathological histology;three pro-in-flammatory factors,namely,TNF-α,IL-10,and IL-1β,were detected by ELISA;and the cyber-pharmacology technology was utilized by PubChem,TCMSP,STRING,Cytoscape,AutoDocks and other databases and software to construct the PPI network as well as to perform GO function and KEGG enrichment analyses;and molecular docking technology was used to assess the binding ac-tivity of the drugs to the core targets and to speculate on the signaling pathways of the drug action.The results showed that BC had a better inhibitory effect on KP in the in vitro experiments;path-ological histology showed a significant therapeutic effect of BC;compared with the infected group,the content of pro-inflammatory factors TNF-α,IL-10,and IL-1β in the baicalin treatment group were significantly decreased(P≤0.05).Twenty-four core targets and 11 pathways of action were screened by network pharmacology,and BC docked stably with the acquired core targets TP53,PTGS2,MAPK1,MAPK8,TNF,BCL2,and IGF1 molecules,and it was speculated that BC might exert its antibacterial and anti-inflammatory effects through the signaling pathways of PI3K-Akt,MAPK,HIF-1,and NF-kappa B,etc.This study lays the foundation for further research on the mechanism of action of baicalin on renal injury.

Objective:To analyze the efficacy of endovascular treatment for venous outflow tract obstruction after liver transplantation.Methods:A retrospective analysis was conducted on the data of 7 patients with venous outflow tract obstruction after liver transplantation admitted to Beijing Friendship Hospital, Capital Medical University from November 2020 to December 2024. Among them, there were 5 males and 2 females, with the age of (22.3±8.1) years. The primary diseases included 2 cases of Budd-Chiari syndrome, 1 case of hepatic veno-occlusive disease, 1 case of portal veno-hepatic sinus vascular disease, 1 case of ornithine carbamoyltransferase deficiency, 1 case of primary biliary cirrhosis, and 1 case of autoimmune cirrhosis. Analyze the patient's clinical manifestations, obstruction of venous outflow tract, hemoglobin levels within one week before and one week after the operation, endovascular treatment conditions, and intraoperative complications such as abdominal hemorrhage and vascular injury. Ultrasound was used to measure the depth of ascites and pleural effusion. All patients were followed up immediately after the operation through phone calls or follow-up visits. The clinical symptoms, abdominal vascular ultrasound, enhanced CT and survival status of the patients were followed up.Results:All 7 patients were diagnosed with venous outflow tract obstruction by intraoperative angiography, including 1 cases of inferior vena cava obstruction, 2 cases of hepatic vein obstruction, and 4 cases of vena cava combined with hepatic vein obstruction. A total of 12 endovascular treatments were performed on 7 patients. Among them, 4 patients received balloon dilation and angioplasty once, 1 patient received balloon dilation and angioplasty twice, 1 patient underwent hepatic vein stent implantation after 2 hepatic vein balloon dilation and angioplasty, and 1 patient underwent intrahepatic portosystemic shunt via jugular vein after 2 hepatic vein balloon dilation and angioplasty. The abdominal distensionof the patients were all relieved after the operation, the ascites and pleural effusion decreased, and the edema symptoms of the lower extremities disappeared. There were no intraoperative complications. The preoperative hemoglobin of 7 patients was (113.4±34.0) g/L, and the postoperative hemoglobin was (126.6±34.8) g/L, which increased significantly compared with that before the operation, and the difference was statistically significant ( t=-0.71, P=0.038). Seven patients were followed up for 6 to 24 months, with a median of 12 months. None of them had obvious symptoms including abdominal distension. Abdominal ultrasound and CT indicated that the blood flow of the transplanted liver was unobstructed, and no patient died. Conclusion:Venous outflow tract obstruction after liver transplantation can cause severe symptoms. Endovascular treatment is an effective treatment for venous outflow tract obstruction after liver transplantation.

To analyze the epidemiological characteristics of human infection with avian influenza A(H7N9) virus from 2013 to 2016 in Xiaoshan,China,we collected the epidemiological data of human A(H7N9) cases as well as the results of environmental monitoring and analyzed it by case study,descriptive epidemiological method,and statistical analysis.Results showed that a total of 22 cases were confirmed,including 12 deaths.All the cases occurred in winter and spring,and most of them had been exposed to live poultry,and with the underlying condition of chronic diseases.Chi-square test showed that the ratio of cases with chronic diseases in death group was higher (P=0.01).The environmental monitoring result showed there were distinctive seasonal differences of positive rate with peaks in winter and spring and nadirs in autumn and summer.Positive rates for the samples from external environmental monitoring were consistent with the distribution of the case onset.Due to rise of environmental monitoring positive rate,we should be alert to the occurrence of new cases,especially in rural areas where live poultry fairs still exist and among people with occupational exposure.We should further improve ability to prevent and control this disease.

To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.