Efficacy and safety of sofosbuvir/velpatasvir alone or in combination with ribavirin in treatment of patients with genotype 3B HCV/HIV infection / 临床肝胆病杂志

ObjectiveTo investigate the efficacy and safety of sofosbuvir/velpatasvir alone or in combination with ribavirin in Chinese patients with genotype 3B HCV/HIV infection. MethodsA total of 299 patients with genotype 3B HCV/HIV infection who attended The Third People’s Hospital of Kunming from January 2017 to December 2020 were enrolled and treated with sofosbuvir/velpatasvir alone or in combination with ribavirin for 12 weeks， and they were followed up for 12 weeks after drug withdrawal. The patients were evaluated in terms of sustained virologic response at 12 weeks after treatment （SVR12） and adverse reactions. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups， and the chi-square test was used for comparison of categorical data between two groups； the Agresti-Coull method was used to evaluate the 95% confidence interval （CI） of SVR12； univariate and multivariate non-conditional logistic regression analyses were used to investigate the influencing factors for SVR. ResultsThe 299 patients with genotype 3B HCV/HIV infection had a mean age of 43.92±6.84 years， among whom the male patients accounted for 77.3% （231/299）， the patients with liver cirrhosis accounted for 36.5% （109/299）， the patients with a history of antiviral therapy accounted for 13.4% （40/299）， and the patients receiving sofosbuvir/velpatasvir combined with ribavirin accounted for 27.8% （83/299）. The overall SVR was 87.0% （260/299） for all patients， and there was no significant difference in SVR12 between the patients receiving sofosbuvir/velpatasvir alone and those receiving sofosbuvir/velpatasvir combined with ribavirin （87.5% vs 85.5%， χ2=0.203， P=0.653）. There was a significant difference in SVR12 between the patients without liver cirrhosis and those with liver cirrhosis （90.0% vs 81.7%， χ2=4.256， P=0.039）， and the patients receiving antiviral therapy for the first time had a significantly higher SVR12 than the treatment-experienced patients （93.4% vs 45.0%， χ2=71.670， P<0.001）. The univariate and multivariate logistic regression analyses showed that platelet count （odds ratio ［OR］=0.957， 95%CI： 0.931 — 0.984， P=0.002）， liver stiffness measurement （OR=1.446， 95%CI： 1.147 — 1.822， P=0.002）， and experience in treatment （OR=13.807， 95%CI： 2.970 — 64.174， P=0.001） were independent influencing factors for SVR in patients with genotype 3B HCV/HIV infection. There were 41 cases of serious adverse events， all of which occurred within 2 weeks after antiviral therapy， and 28 cases were resolved without drug withdrawal or active treatment， while 13 cases were not resolved after active treatment and were resolved after the antiviral drugs were stopped for 2‍ ‍—‍ ‍5 days， with no similar reactions observed when the drugs were used again after remission. ConclusionSofosbuvir/velpatasvir alone or in combination with ribavirin has relatively good efficacy and safety in patients with genotype 3B HCV/HIV infection.

Efficacy and safety of sofosbuvir-based direct-acting antiviral treatment in children and adolescent patients with chronic hepatitis C / 中华传染病杂志

Objective:To explore the efficacy and safety of sofosbuvir-based direct-acting antiviral treatment in children and adolescent patients with chronic hepatitis C (CHC).Methods:A total of 52 children and adolescent patients who admitted to The Third People′s Hospital of Kunming City and The People′s Hospital of Fuyuan County aged from three to 17 years old with CHC from January 2018 to August 2022 were enrolled, and their basic information was collected. Patients were treated with sofosbuvir/velpatasvir (SOF/VEL) or ledipasvir/sofosbuvir (LDV/SOF) with or without ribavirin for 12 weeks. The biochemical and virological indexes were followed up before and after treatment and 12 weeks after withdrawal. The primary endpoint was the sustained virological response (SVR) at week 12 of follow-up after treatment, and the occurrence of adverse events (AE) during treatment. Statistical analysis was used by nonparametric test.Results:A total of 52 patients with CHC including 38 children and 14 adolescents were enrolled. Thirty-one were male and 21 were female. The age was 9(7, 12) years old. Among 52 patients, seven patients were type 1b, 11 were type 2a, three were type 2, five were type 3a, 18 were type 3b, one was type 6a, three were type 6k, four were type 6n and one was type 6v. Twelve (23.1%) patients were vertical transmission, 21(40.4%) patients had horizontal transmission among family members, two (3.8%) patients were blood fluid transmission, and 17(32.7%) were unknown transmission route. Compared with the baseline levels, Total bilirubin, alanine aminotransferase and aspartate aminotransferase were all significantly decreased after 12 weeks of treatment and 12 weeks after withdrawal, and the differences were statistically significant ( F=12.71, 30.23 and 42.52, respectively, all P<0.05). Up to September 30, 2022, 100.0%(52/52) of patients achieved SVR at the end of treatment. For patients who completed follow-up for 12 weeks after treatment, 95.8%(46/48) achieved SVR. Common AEs during treatment were fatigue (11.5%(6/52)), headache (5.8%(3/52)), dizziness (1.9%(1/52)), abdominal pain (3.8%(2/52)), diarrhea (1.9%(1/52)), rash (1.9%(1/52)) and skin pruritus (1.9%(1/52)). No patients discontinued treatment because of AE. Conclusions:Sofosbuvir-based direct-acting antiviral treatment is efficient and well-tolerated in children and adolescent patients with CHC. No patients discontinued treatment due to AE.

Establishment and evaluation of a multivariate Cox proportional-hazards prediction model for mortality during short-term hospitalization in patients with liver cirrhosis and sepsis / 临床肝胆病杂志

Objective To establish a Cox proportional-hazards prediction model for mortality during short-term hospitalization in patients with liver cirrhosis and sepsis. Methods A retrospective analysis was performed for the clinical data of 336 patients with liver cirrhosis and sepsis who were admitted to The Third People's Hospital of Kunming from January 2012 to August 2022, and according to whether the patient died during short-term hospitalization, they were divided into death group with 40 patients and survival group with 296 patients. Demographic data, comorbidities, and clinical biochemical parameters were collected and compared between the two groups. The independent-samples t test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The multivariate Cox analysis was used for screening of variables, then a Cox proportional-hazards prediction model was established, and hazard ratio ( HR ) and its 95% confidence interval [ CI ] were calculated; C-index index was used to evaluate the prediction accuracy of the model. The Cox proportional-hazards prediction model was visualized by a nomogram, and calibration curve was plotted to evaluate the consistency between the prediction results of the model and the actual condition. Results Among the 336 patients, there were 261 male patients (77.7%) and 75 female patients (22.3%), with a mean age of 50.0±10.6 years, and 40 patients died, with a mean hospital stay of 16.8±11.3 days (range 8.2-23.0 days). Compared with the survival group, the death group had a significantly higher proportion of patients with an age of ≥60 years, a history of invasive operation within the past two weeks, gastrointestinal bleeding, hepatic encephalopathy (HE) or hepatorenal syndrome (HRS), a significantly higher Modified Early Warning Score (MEWS) score, and significantly higher levels of prothrombin time (PT), activated partial thromboplastin time, international normalized ratio, D-dimer, CD4/CD8 ratio, lactate, white blood cell count, norepinephrine, total bilirubin, interleukin-6, procalcitonin, high-sensitivity C-reactive protein (hsCRP), blood urea nitrogen, and creatinine (all P < 0.05), as well as significantly lower levels of red blood cell count, hemoglobin, albumin, total cholesterol, low-density lipoprotein, and high-density lipoprotein (all P < 0.05). The multivariate Cox regression analysis showed that age ( HR =2.602, 95% CI : 1.277-5.303, P =0.008), HE ( HR =2.516, 95% CI : 1.258-5.033, P =0.009), HRS ( HR =2.324, 95% CI : 1.010-5.349, P =0.047), hsCRP ( HR =1.008, 95% CI : 1.003-1.013, P =0.004), MEWS score ( HR =1.205, 95% CI : 1.022-1.422, P =0.027), and PT ( HR =1.076, 95% CI : 1.030-1.124, P =0.027) were independent influencing factors for death in patients with liver cirrhosis and sepsis. The model showed a C-index of 0.857 (95% CI : 0.815-0.920), suggesting that the model had relatively high prediction accuracy, and the calibration curve showed good consistency between the predicted risk and the actual risk. Conclusion The Cox proportional-hazards prediction model established for death during short-term hospitalization in patients with liver cirrhosis and sepsis can be used to predict the risk of death during short-term hospitalization in patients with liver cirrhosis and sepsis, thereby guiding clinical medical staff to take targeted intervention measures to avoid or reduce the possibility of death in patients.

Effect of Ganshuang granule combined with entecavir on portal vein thrombosis in patients with hepatitis B cirrhosis / 临床肝胆病杂志

Objective To investigate the effect of Ganshuang granule combined with entecavir on portal vein thrombosis (PVT) in patients with hepatitis B cirrhosis. Methods A total of 356 patients with hepatitis B cirrhosis who attended and were hospitalized in The Third People's Hospital of Kunming from January 1, 2018 to December 31, 2020 were enrolled and randomly divided into combination group with 191 patients and control group with 165 patients. The patients in the combination group received Ganshuang granule combined with entecavir, and those in the control group received entecavir alone. The course of treatment was at least 24 weeks. The t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups. The Kaplan-Meier method was used to analyze the cumulative incidence rate of PVT in both groups, and the log-rank test was used for comparison between two groups. Univariate and multivariate Cox proportional-hazards regression model analyses were used to investigate the influencing factors for PVT in patients with hepatitis B cirrhosis. Results The 191 patients in the combination group were followed up for 296.25 person-years in total, with a mean follow-up time of 1.55±0.65 years, and there were 8 patients with PVT, with an incidence rate of 4.19% and an incidence density of 1.41 per ten-thousand person-years. The 165 patients in the control group were followed up for 253.25 person-years in total, with a mean follow-up time of 1.53±0.67 years, and there were 20 patients with PVT, with an incidence rate of 12.12% and an incidence density of 4.79 per ten-thousand person-years. There was a significant difference in the incidence rate of PVT between the two groups ( χ 2 =7.687, P =0.006). The cumulative incidence rate of PVT plotted by the Kaplan-Meier method showed that the combination group had a significantly lower cumulative incidence rate of PVT than the control group ( χ 2 =7.226, P =0.007), with a relative risk of 3.155 (95% confidence interval [ CI ]: 1.351-7.370). The univariate Cox analysis showed that hypertension, alanine aminotransferase, aspartate aminotransferase (AST), albumin (Alb), cholinesterase, estimated glomerular filtration rate, alpha-fetoprotein, D-dimer (D-D), Child-Pugh class, and Ganshuang granule combined with entecavir were influencing factors for PVT (all P < 0.05); the multivariate Cox analysis showed that AST (hazard ratio [ HR ]=1.002, 95% CI : 1.000-1.004, P =0.025), and D-D ( HR =1.907, 95% CI : 1.554-2.338, P < 0.001) were independent risk factors for PVT in patients with hepatitis B cirrhosis, while Alb ( HR =0.844, 95% CI : 0.755-0.944, P =0.003) and Ganshuang granule combined with entecavir ( HR =0.350, 95% CI : 0.144-0.851, P =0.021) were independent protective factors against PVT in patients with hepatitis B cirrhosis. Conclusion Ganshuang granule combined with entecavir can significantly reduce the incidence rate of PVT in patients with hepatitis B cirrhosis, thereby exerting a certain preventive effect against PVT.

Influencing factors for liver inflammation and fibrosis in the immune-tolerant phase of HBV infection / 临床肝胆病杂志

Objective To evaluate the degree of liver injury and liver fibrosis in patients in the immune-tolerant phase of chronic HBV infection, and to provide a basis for judging the condition of patients in the immune-tolerant phase. Methods A total of 300 patients with HBV DNA ≥10 7 IU/mL, alanine aminotransferase (ALT) ≤40 U/L, and complete data who were treated in The Third People's Hospital of Kunming from January 2015 to December 2019 were enrolled as subjects, and related data were collected, including age, sex, duration of HBV infection, blood biochemistry, hepatitis B surface antigen (HBsAg) level, and HBV DNA. Liver pathological examination was performed for all patients, and the patients were divided into G < 2 and G ≥2 groups according to inflammation grade and S < 2 and S ≥2 groups according to the degree of fibrosis. The t -test was used for comparison of continuous data between two groups, and univariate and multivariate unconditional logistic regression analyses were used to investigate the influencing factors for G ≥2 liver inflammation and S ≥2 liver fibrosis. Results Among the 300 patients, 213 (71%) had G ≥2 liver inflammation and 120 (40%) had S ≥2 liver fibrosis, with a baseline age of 26.06±9.01 years; male patients accounted for 48%, and the duration of infection was 5.62±5.09 years. The univariate analysis showed that there were significant differences between the G < 2 and G ≥2 groups in ALT, alkaline phosphatase (ALP), albumin (Alb), platelet count (PLT), diameter of the portal vein, and spleen thickness ( t =-26.677, -11.612, 2.149, 5.410, -6.092, and -2.911, all P < 0.05), and there were significant differences between the S < 2 and S ≥2 groups in duration of infection, ALT, ALP, Alb, HBV DNA, PLT, diameter of the portal vein, and spleen thickness ( t =-6.320, -6.694, -7.880, 2.349, 4.552, 19.160, -5.782, and -5.622, all P < 0.05). The multivariate analysis showed that ALT (odds ratio [ OR ]=10.270, 95% confidence interval [ CI ]: 2.212-47.672, P =0.003) and ALP ( OR =1.097, 95% CI : 1.013-1.188, P =0.023) were independent risk factors for G ≥2 liver inflammation in patients in the immune-tolerant phase, and ALP ( OR =1.034, 95% CI : 1.015-1.054, P < 0.001), PLT ( OR =0.913, 95% CI : 0.886-0.938, P < 0.001), HBV DNA ( OR =0.198, 95% CI : 0.062-0.636, P =0.007), and duration of infection ( OR =1.176, 95% CI : 1.033-1.340, P =0.015) were independent influencing factors for S ≥2 liver fibrosis in patients in the immune-tolerant phase. Conclusion Most patients in the immune-tolerant phase have significant liver histological changes. ALT and ALP are the influencing factors for significant liver inflammation, and ALP, HBV-DNA, PLT, and infection time are the influencing factors for significant liver fibrosis in patients in the immune-tolerant phase.

Liver inflammatory activity in chronic hepatitis B patients with a low HBV DNA level and related factors / 临床肝胆病杂志

ObjectiveTo investigate the association between liver inflammatory activity and clinical features in chronic hepatitis B (CHB) patients with a low HBV DNA level, and to provide a basis for judging the condition of such patients. MethodsA total of 137 patients with an HBV DNA level of 20-2000 IU/ml who were treated in The Third People′s Hospital of Kunming from January 2014 to December 2017 were enrolled, and according to the grade of liver inflammatory activity, they were divided into G1 group with 44 patients, G2 group with 84 patients, and G3 group with 9 patients. Liver biopsy and immunohistochemistry were performed for all patients, and the association of the grade of liver inflammatory activity with age, sex, HBV infection time, liver function, HBsAg level, HBV DNA load, genotype, diameter of the portal vein, diameter of the splenic vein, and spleen thickness was analyzed. The Kruskal-Wallis H test was used for comparison of nonparametric ranked data between multiple groups. ResultsOf all 137 patients, 126 had negative HBeAg, among whom 42 (33.33%) had G1 inflammation, 77 (61.11%) had G2 inflammation, and 7 (5.56%) had G3 inflammation; 11 had positive HBeAg, among whom 2 (18.18%) had G1 inflammation, 7 (63.64%) had G2 inflammation, and 2 (18.18%) had G3 inflammation. There were significant differences in globulin, HBV DNA load, platelet count (PLT), diameter of the splenic vein, and spleen thickness between the 126 HBeAg-negative patients with different inflammation grades (H=6.189, 7.168, 8.624, 8.170, and 8.522, all P＜0.05). ConclusionThe majority of patients with a low HBV DNA level have ≥G2 liver inflammation and the indication for antiviral therapy, and the grade of inflammation is closely associated with globulin, HBV DNA load, PLT, diameter of the splenic vein, and spleen thickness.

Effects of astragaloside Ⅳ on inflammatory responses in astrocytes induced by lipopolysaccharide in vitro and its mechanism / 中华微生物学和免疫学杂志

Objective To investigate whether astragaloside ( AST) Ⅳ could inhibit lipopolysac-charide (LPS)-induced activation of astrocytes and the possible mechanism. Methods Effects of different concentrations of AST Ⅳ on astrocyte viability were determined by MTT to select the optimum concentration for the following experiments. Primary astrocytes were induced by LPS to construct the inflammatory model. Astrocytes were divided into three groups: PBS, LPS and LPS+ASTⅣ(LPS+ASTⅣ) groups. The release of nitric oxide (NO) was detected by Griess method. Expression of glial fibrillary acidic protein ( GFAP), an astrocyte marker, and TLR4 were analyzed by immunohistochemistry and Western blot. Cytokines of IL-6, TNF-α, IL-4 and IL-10 in the supernatants of cell culture for 24 h collected after stimulation were meas-ured by ELISA. Results ASTⅣsignificantly inhibited the LPS-induced activation of astrocytes and NO re-lease (P<0. 01), suppressed the expression of TLR4 (P<0. 05) and reduced the secretion of IL-6 (P<0. 01) and TNF-α (P<0. 01), but increased the secretion of IL-4 and IL-10 (P<0. 05 and P<0. 01). Con-clusion AST Ⅳ could significantly inhibit the LPS-induced activation of astrocytes and suppress inflamma-tory responses, and the possible mechanism might be related to reduced secretion of inflammatory factors af-ter blocking the expression of TLR4.

Correlations betweenintrahepatic HBsAg and HBcAg levels and serum HBsAg and HBV DNA levels in low-level HBsAg patients / 实用医学杂志

Objective To analyze the relationship between intrahepatic expression of HBsAg and HBcAg and the serum HBsAg and the load of HBV DNA in low-level HBsAg.To provide the basis for the diagnosis of low-level HBsAg.Methods A total of 63 patients with HBsAg lower than 1 400 IU/mL and higher than 0.05 IU/mL were enrolled in the study.All the 63 patients with low level HBsAg were collected for liver biopsy from January 2013 to December 2015.Serum HBsAg was detected by Roche cobas e601 automatic electrochemiluminescence immunoas-say system and supporting reagents.HBV DNA was detected using ROCHE automatic AmpliPrep/cobas TaqMan 48 system detector and supporting reagents.Results Among the 63 patients,the HBsAg in liver tissue was negative in 5 cases(7.94%),+in 47 cases(74.60%),++in 8 cases(12.70%),+++in 2 cases(3.17%),and++++was found in 1 case.The expression of HBcAg were negative in 30 cases(47.62%),+in 28 cases(44.44%),++in 4 cases(6.35%),+++in 1 case(1.59%),and++++in 0 cases.There was a significant positive correlation between HBsAg expression and serum HBsAg level by spearman and kendall correlation analysis(spearman analysis:r = 0.261,P=0.039;kendall analysis:r=0.217,P=0.036).HBcAg expression was significantly correlated with serum HBV DNA load with spearman and kendall correlation analysis(spearman analysis:r=0.305,P=0.015;kendall analysis:r=0.259,P=0.017).Conclusions The expression of HBsAg in liver tissue of in the patients with low level of HBsAg is positively correlated with the level of serum HBsAg,but not with the load of HBV DNA. There is significant positive correlation between HBcAg expression and serum HBV DNA load.

Immunoregulatory effect of BuyangHuanwu Decoction on experimental autoimmune encephalomyelitis mice / 中国免疫学杂志

Objective:To explore the anti-inflammatory therapeutic effect and possible immunoregulatory mechanism of Buyang Huanwu Decoction (BYHWD) on the development of experimental autoimmune encephalomyelitis (EAE). Methods:Female C57BL/6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptides ( MOG35-55 ) ,and randomly divided into saline group,BYHWD group,with 13 mice in each group. At the 3th day,25 ml/kg of saline was orally given to each mouse of saline group,50 g/kg ig of crude BYHWD was orally given to each mouse in BYHWD group for 25 days. Clinical score and body mass were recorded every day. Inflammatory cell infiltrations of spinal cord were observed by HE staining Myelin staining observes the demyelination situa-tion. And the expression of ROCKⅡ in spleen was detected by immunofluorescence staining. The subtypes of CD4+ T cells were analyzed by flow cytometry. Western blot was used to detect the expression of TLR4,Myd88,NF-κB,COX-2,ROCKⅡ in spinal cord and ROCKⅡ in brain. Results: The neurologicalscore significantly decreased in EAE mouse of BYHWD group compared with the saline group (P<0. 001) . BYHWD inhibited the inflammatory cell infiltration and demyelination in the nervous centralis(P<0. 05). The treatment of BYHWD effectively reduced the increased the proportion of CD25+(P<0. 05),IL-10+(P<0. 05),TGF-β+(P<0. 01), IFN-γ+( P<0. 05 ) CD4+T cells , and inhibited the expression of peripheral and central ROCKⅡ( P<0. 05 ) ;BYHWD reduced the expression of TLR4,MyD88,NF-κB,COX-2 in spinal cord (P<0. 05). Conclusion:BYHWD can exert anti-inflammatory and immune regulation effect by the inhibition of ROCKⅡ/TLR4/ NF-κB signaling pathway and regulation of the proportion of peripheral T cell sub-sets.

Immunoregulatory effect of Buyang-Huanwu decoction on monocyte-mac-rophages in mice with experimental autoimmune encephalomyelitis / 中国病理生理杂志

AIM:To explore the therapeutic effect of Buyang-Huanwu decoction (BYHWD) on experimental au-toimmune encephalomyelitis ( EAE) and its immunoregulatory effect on monocyte-macrophages .METHODS: Chronic EAE was induced by myelin oligodendrocyte glycoprotein peptide fragment 35-55 ( MOG35-55 ) in the female C57BL/6 mice, which were randomly divided into saline group and BYHWD group .On day 3 after immunization , the mice in BYHWD group were orally administrated with BYHWD , while normal saline was given to the control mice .The clinical score and body mass were recorded every other day .At day 17 after immunization , the mice were sacrificed and spinal cords were obtained for HE staining and myelin staining .The M1 and M2 macrophage phenotypes of splenic cells were detected by flow cytometry and immunofluorescence staining .The protein expression of iNOS , TNF-α, arginase and IL-10 in the spinal cord macro-phages was determined by Western blotting .RESULTS:BYHWD delayed the onset of EAE , reduced the clinical scores of EAE, inhibited the inflammatory cell infiltration and demyelination in the spinal cord , and promoted the conversion of M 1 macrophages into M2 phenotype in the spinal cord and spleen .CONCLUSION:BYHWD intervention attenuates the be-havioral and pathological changes in the EAE mice , and its mechanism may be related to the macrophage conversion .

Immunoregulative effect of Fasudil on encephalomyelitic T cells in experimental autoimmune encephalomyelitis mice / 中南大学学报(医学版)

OBJECTIVE@#To explore the therapeutic effect of Fasudil-modified splenic mononuclear cells (MNCs) in experimental autoimmune encephalomyelitis (EAE) and the possible mechanisms.
@*METHODS@#C57BL/6 female mice were immunized with myelin oligodendrocyte glycoprotein peptide 35-55 to establish active immunity EAE model. Splenic MNCs were isolated on the 9th day after immunization and treated with or without Fasudil for 72 h in vitro. These cells were collected for analysis of the variance of T cell subtypes, the level of cytokines and the activity of Rho kinase (ROCK). MNCs (5×107 cells) were resuspended in 500 µL of phosphate buffer solution (PBS) and transferred into EAE model (intraperitoneal injection), which was divided into a PBS-MNCs group and a Fasudil-MNCs group. Changes of body weight and clinical symptom scores were observed.
@*RESULTS@#Splenic encephalitogenic MNCs from EAE mice on the 9th day after immunization could establish passive transfer EAE model. But Fasudil-treated MNCs did not trigger EAE development. Compared with the PBS-MNCs group, the loss of body weight was less in the Fasudil-MNCs group. The in vitro experiment indicated that Fasudil could suppress the activity of ROCK on MNCs (P<0.01), decrease the percentage of CD4+ T cells with the expression of interferon-γ (IFN-γ) and interleukin-17 (IL-17) (IFN-γ: P<0.01; IL-17: P<0.05), while increase the secretion of CD4+ T cells with the expression of transforming growth factor-β (TGF-β) and IL-10 (all P<0.001) . Furthermore, Fasudil could inhibit the release of IL-17 (P<0.001) and enhance the level of IL-10 (P<0.05).
@*CONCLUSION@#Fasudil-modified cell therapy affects the occurrence and development of EAE by inhibiting the inflammatory reaction of helper T cell 1 (Th1) and Th17 while enhancing the immunoregulative effect of Th2.

The immunoregulatory effect of Rho kinase inhibitor Fasudil on macrophages in a mouse model of ;experimental autoimmune encephalomyelitis / 中华微生物学和免疫学杂志

Objective To investigate the immunoregulatory effect of Fasudil-modified macrophages on cell transferred experimental autoimmune encephalomyelitis ( EAE) in a mouse model.Methods Fe-male C57BL/6 mice were immunized with MOG35-55 to establish the model of EAE.The encephalomyelitic mononuclear cells ( MNCs) were isolated from spleen of mice with EAE on day 9 after immunization and treated with or without Fasudil for 72 h in vitro.Several assays including the flow cytometry analysis, Griess reaction and ELISA were performed to analyze the M1 and M2 phenotypes of macrophages, the production of NO and the levels of cytokines, respectively.The cultured MNCs (5×107 cells) were resuspended in 500μl of PBS and transferred into na?ve C57BL/6 recipients via intraperitoneal injection.Two groups including the PBS-MNCs group and the Fasudil-MNCs group were set up.The body weights and clinical scores of the mice in each group were recorded in every other days after the induction of EAE in the recipients.Results The Fasudil treated MNCs affected the induction of EAE in adoptive cell transferred mice.The expression of CD16/32, iNOS and IL-12 on F4/80-macrophages were decreased, while the expression of CD206, CD23 and IL-10 on F4/80-macrophages were increased upon the treatment of Fasudil, indicating that Fasudil im-proved the differentiation of macrophages from M1 to M2 phenotypes.Moreover, Fasudil inhibited the pro-duction of NO and enhanced the expression of Arginase-1.Conclusion Fasudil ameliorated the clinical se-verity of EAE in mice by promoting the transformation of macrophages from M1 to M2 phenotype.

Lipoic acid protects dopaminergic neurons in LPS-induced model of Parkinson’s disease / 中国病理生理杂志

AIM:To evaluate the effect of lipoic acid ( LA) on LPS-induced Parkinson disease ( PD) model of mice.METHODS:Female C57BL/6 mice of 10-month-old were randomly divided into saline control group , PD group and LA group.The PD mouse model was induced by intranasal instillation of LPS .Assays of tyrosine hydroxylase , microglia and nuclear factor kappa B ( NF-κB) were performed by the methods of immunohistochemistry and Western blotting .RE-SULTS:Intranasal LPS instillation exhibited basic characteristics of PD model .However, LA administration significantly improved motor dysfunction , protected dopaminergic neurons from damage , and inhibited NF-κB activation in inflammatory microglia in the substantia nigra area of the brain .CONCLUSION:LA may exert a profound neuroprotective effect by an-ti-neuroinflammatory reaction to arrest the progression of PD .

Study on a novel Rho kinase inhibitor WAR5 for treating EAE / 中国病理生理杂志

AIM:To explore the therapeutic effect of a novel Rho kinase inhibitor WAR 5 on the experimental autoimmune encephalomyelitis (EAE) and its possible mechanism.METHODS: Female C57BL/6 mice were randomly divided into EAE group and WAR5 group.EAE model was induced by the application of MOG 35-55 peptide.WAR5 was in-jected intraperitoneally every other day from post-immunization (PI) day 3 to PI day 27.The clinical score and body weight were recorded every other day .On PI day 28, the animals were sacrificed and spinal cords were obtained for HE and mye-lin staining .The splenocytes were isolated and the expression of CD 16/32 and CD206 were analyzed by flow cytometry . The protein extracts from the brains and spinal cords were collected for the measurement of inducible nitric oxide synthase ( iNOS) by Western blotting .RESULTS:The administration of WAR 5 delayed the onset of EAE and attenuated the clini-cal symptoms .The results of the pathological examination revealed that WAR 5 inhibited the infiltration of inflammatory cells and improved myelination in spinal cords , accompanied with the poralization of M 1 macrophages to M2 phenotype in the spleen.WAR5 inhibited the expression of iNOS in the central nervous system , especially in the spinal cords .CON-CLUSION:The therapeutic effect of WAR5 on EAE may be related to the shift of M1 macrophages to M2 phenotype and inhibition of inflammation in the central nerve system .