Objective To investigate the application value of tenofovir alafenamide fumarate(TAF)in patients with first-time hepatitis B virus-related decompensated cirrhosis(HBV-DC)and its impact on renal function and lipid metabolism.Methods A total of 57 patients with first-time HBV-DC who were hospitalized and received TAF antiviral therapy in The Affiliated Hospital of Xuzhou Medical University from January 1,2020 to December 31,2022 were enrolled,and all of them received TAF antiviral therapy.Related data were collected at baseline and at weeks 12,24,and 48 of treatment,including virological and serological indicators,liver and renal function,serum phosphorus,and blood lipids.The paired t-test or single group repeated measures ANOVA were used for comparison of normally distributed continuous data,the Friedman test was used for comparison of non-normally distributed continuous data,and the chi-square test or the Fisher's exact test were used for categorical data.Results A total of 52 patients completed the 48 weeks of follow-up.After 12,24,and 48 weeks of treatment,the patients achieving HBV DNA seroconversion accounted for 38.5%,63.5%,and 84.6%,respectively;the alanine aminotransferase normalization rate were 71.2%,82.7%,and 82.7%,respectively;the proportion of the patients with Child-Pugh class A disease increased to 55.8%,73.1%,and 92.3%,respectively.Within the 48 weeks of treatment,there were significant increases in the levels of cystatin C(χ2=35.163,P＜0.001)and serum phosphorus(F=8.600,P＜0.001)and low-density lipoprotein cholesterol(χ2=10.064,P=0.018).The ratio of total cholesterol/high-density lipoprotein cholesterol decreased continuously from 3.61(2.61～5.84)to 3.27(2.70～4.36)(χ2=5.000,P=0.172).Conclusion TAF can rapidly inhibit HBV replication and significantly improve liver function in HBV-DC patients,with no significant impact on renal function.However,blood lipid should be closely monitored.

Objective:To study the clinical features and prognostic impact of transarterial chemoembolization (TACE), immune checkpoint inhibitors (ICIs), and tyrosine kinase inhibitors (TKIs) combination therapy regimens in the treatment of patients with hepatitis B virus-related intermediate-and advanced-stage hepatocellular carcinoma with secondary cholestasis.Methods:Patients with HBV-related intermediate-and advanced-stage hepatocellular carcinoma (HBV) who visited the Affiliated Hospital of Xuzhou Medical University between January 1, 2020, and December 31, 2022, were enrolled. TACE+TKIs +ICIs combination therapy was used to treat all patients. The occurrence and factors influencing cholestasis, as well as the impact on prognosis after combined therapy, were analyzed. The measurement data were compared using a t-test and a non-parametric rank sum test. The count data was compared using the χ2 test. The survival rates were compared using a log-rank test between different groups. Results:A total of 106 cases with HBV-related intermediate-and advanced-stage hepatocellular carcinoma were enrolled. The probabilities of secondary cholestasis within 3 and 6 months, 1, 2, and 3 years after TACE+ICIs+TKIs combination therapy were 9.4%, 12.3%, 14.2%, 24.5%, and 24.5%, respectively. Patients with secondary cholestasis had persistent symptoms and rapid progression. During the treatment course, the median survival time was significantly longer in patients with hepatocellular carcinoma without secondary cholestasis than that of patients with cholestasis (26.9 months vs. 13.7 months, respectively, P < 0.05). Secondary cholestasis, baseline aspartate aminotransferase, and prothrombin activity levels were independent risk factors that affected the survival and prognosis of patients treated with combination therapy. There was no statistically significant difference in the occurrence of other adverse reactions between the two groups with secondary and non-secondary cholestasis during the treatment course (47.5% vs. 43.3%, χ2=0.058, P = 0.810). Conclusion:TACE+ICIs+TKIs therapy combination is relatively common in the treatment of patients with HBV-related intermediate-and advanced-stage hepatocellular carcinoma with secondary cholestasis. Moreover, accelerated disease progression is an independent risk factor affecting the survival and prognosis of patients.

ObjectiveTo investigate the effect of HBsAg on the expression of interferon-α (IFN-α) in peripheral blood plasmacytoid dendritic cells (pDCs) induced by the stimulator of interferon genes (STING) signaling pathway activated by cyclic GMP-AMP (cGAMP). MethodPeripheral venous blood was collected from healthy adults and the patients with chronic hepatitis B virus (HBV) infection who attended the outpatient service or were hospitalized in Department of Infectious Diseases, The Affiliated Hospital of Xuzhou Medical University, from February to December 2016, and peripheral blood mononuclear cells (PBMCs) were isolated and extracted. After the STING agonist cGAMP was added to PBMCs, ELISA was used to measure the levels of IFN-α, interferon-β, and tumor necrosis factor-α in supernatant. PBMCs from healthy adults were pre-incubated with HBsAg and then stimulated by cGAMP, and supernatant was collected to measure IFN-α. The magnetic-activated cell sorting method was used to remove pDCs from PBMCs, and after culture with cGAMP, ELISA was used to measure the level of IFN-α in supernatant. PBMCs from healthy adults were stimulated by HBsAg and/or cGAMP, and then flow cytometry was used to measure the frequency of pDCs. The independent samples t-test was used for comparison of continuous data between two groups. ResultsPBMCs from the patients with chronic HBV infection stimulated by cGAMP in vitro had a significantly lower level of IFN-α than healthy controls (469.72±18.95 vs 599.90±84.06, t=4.868, P=0.001). PBMCs from healthy adults co-cultured with HBsAg and stimulated by cGAMP had a significantly lower level of IFN-α than those in the non-HBsAg group (448.5±52.0 vs 571.0±30.8, t=4.500, P=0.011). Compared with PBMCs containing pDCs, PBMCs without pDCs stimulated by cGAMP had a significant reduction in the level of IFN-α (164.50±40.73 vs 339.50±35.33, t=6.482, P=0.001). Compared with PBMCs from healthy adults stimulated by cGAMP, PBMCs pre-incubated with HBsAg and then stimulated by cGAMP had a significant reduction in the frequency of pDCs (0.12%±0.04% vs 0.24%±0.04%, t=5.176, P=0.014). ConclusionHBsAg can inhibit the expression of IFN-α induced by the STING pathway in pDCs activated by cGAMP.

ObjectiveTo investigate the influencing factors for persistent low-level viremia (LLV) in chronic hepatitis B(CHB) patients receiving long-term entecavir antiviral therapy. MethodsThe CHB patients who received entecavir antiviral therapy for at least one year in The Affiliated Hospital of Xuzhou Medical University from November 2018 to June 2020 were enrolled as subjects, and according to HBV DNA load at the end of the observation period, the patients were divided into LLV group and sustained virological response (SVR) group. Demographic features and laboratory markers were observed for all patients. 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. A multivariate logistic regression analysis was used to investigate the influencing factors for LLV in patients receiving long-term entecavir treatment. ResultsA total of 560 CHB patients were enrolled, with 204 in the LLV group and 356 in the SVR group. There were significant differences between the two groups in age (Z=-3.530, P＜0.001), sex (χ2=4.270, P=0.039), presence or absence of liver cirrhosis (χ2=53.879, P＜0.001), medication compliance (χ2=5.326, P=0.021), HBeAg positive rate (χ2=90.681, P＜0.001), baseline HBV DNA load before treatment (Z=-8.337, P＜0.001), baseline HBsAg quantification (Z=-10.472, P＜0.001), and medication type (χ2=7.558, P=0.006). The multivariate logistic regression analysis showed that baseline HBeAg status before treatment (odds ratio ［OR］=3.381, 95% confidence interval ［CI］: 1.985-5.756, P＜0.001), HBV DNA load before treatment (OR=1.223, 95%CI: 1.050-1.424, P=0.010), and HBsAg quantification before treatment (OR=2.448, 95%CI: 1.743-3.438, P＜0.001) were risk factors for LLV in long-term entecavir antiviral therapy. ConclusionIn clinical practice, CHB patients with high HBV DNA load, high HBsAg quantification, and positive HBeAg tend to have a high risk of LLV even after long-term entecavir antiviral therapy. Therefore, such population should be taken seriously with the dynamic monitoring of HBsAg quantification, HBV DNA load, and HBeAg status.

Objective To evaluate the effect of interleukin (IL)-10 on donor lung function after ex vivo lung perfusion (EVLP) in rats of cardiac death. Methods Twenty adult male SD rats were randomly divided into the simple perfusion group (group A, n=10) and modified perfusion group (group B, n=10). Perfusate A (without IL-10) and perfusate B (supplemented with IL-10) was administered in group A and B, respectively. The EVLP rat models of cardiac death were established. The appearance of donor lung, dry-to-wet (D/W) ratio of donor lung tissues, the function and metabolism of donor lung, the morphology of donor lung and the levels of inflammatory markers of donor lung were statistically compared between two groups. Results After perfusion, evident edema of the whole donor lung, poor compliance and a large amount of edema fluid discharged from the airway were observed in group A, whereas no obvious edema and good compliance were found in group B. Compared with group A, the D/W ratio of lung tissues in group B was higher (P < 0.05). In both groups, the pulmonary vein partial pressure of oxygen reached the peak at 2 h after perfusion, which did not significantly differ between two groups (P > 0.05). In group B, the pulmonary artery pressure was increased at a lower speed and significantly lower after perfusion, and the lactic acid level in the perfusate was significantly lower than those in group A (all P < 0.05). In group A, the alveolar structure was largely destroyed and the cells was rare. In group B, the alveolar structure was relatively normal without evident cell edema. The incidence of cell apoptosis of donor lung was high in group A, whereas no obvious cell apoptosis of donor lung was noted in group B. After perfusion for 4 h, the levels of monocyte chemoattractant protein (MCP)-1 and IL-6 were significantly increased, the IL-4 levels were remarkably decreased (all P < 0.05), but the levels of tumor necrosis factor (TNF)-α, IL-1α and inducible nitric oxide synthase (iNOS) did not significantly change in both groups (all P > 0.05). Conclusions IL-10 may improve the function of donor lung after EVLP in rat of cardiac death by reducing cell apoptosis.

Objective To assess the brain development of newborns with cardiac septal defects by MRI.Methods The brain MR images of 150 newborns with cardiac septal defects and 50 normal newborns were analyzed retrospectively.We evaluated the brain development by measuring the four indices of lateral ventricle:anterior horn index (F/F'),body index (D/D'),caudate nucleus index (C/C')and Evans index.Independent samples t test was used to compare the differences between the two groups,and the possible positive diagnostic cut-off points were calculated by using the nonparametric ROC analysis.Results There were no significant differences between the congenital heart disease group and the control group in the two indices:F/F'[(0.301±0.035)vs (0.296±0.031);t=1.035,P>0.05]and Evans index [(0.239±0.052)vs (0.233±0.025);t=0.778,P>0.05].The values of D/D'[(0.261±0.039)vs (0.234±0.032);t=3.873,P<0.05)] and C/C'[(0.138±0.018)vs (0.124±0.015);t=4.479,P<0.05]were significantly higher in the congenital heart disease group than in the control group.In the congenial heart disease group,the area under the ROC curve obtained by D/D'and C/C'were 0.698 and 0.750,respectively.The maximum Yuedeng index corresponding to the D/D'value and the C/C'value were 0.28 and 0.12,respectively. Conclusion The body index(D/D')and the caudate nucleus index(C/C')are sensitive to evaluate the differences of the brain volume between the newborns with cardiac septal defects and the normal newborns.It is helpful to find the abnormal brain volume when the value of D/D'is greater than 0.28 and the value of C/C'is greater than 0.12.

BACKGROUND: Afatinib, a second-generation irreversible epidermal growth factor inhibitor receptor for the development of non-small cell lung cancer and secondary drug resistance, has low bioavailability and adverse reactions due to current oral administration. The aim of this study was to prepare a novel drug delivery system, afatinib liposome, and to establish a method for the determination of encapsulation efficiency. METHODS: Four different preparation methods were used to prepare afatinib liposomes, and the optimal preparation process was determined by comparing the encapsulation efficiency and particle size. RESULTS: It has been verified that sephadex microcolumn centrifugation can be used to purify afatinib liposomes, and UV spectrophotometry can be employed to determine the entrapment efficiency of liposomes. Among different preparation methods, the encapsulation efficiency of afatinib liposomes prepared by ammonium sulfate gradient method was 90.73% and the average particle size was 108.6 nm. CONCLUSIONS Ammonium sulfate gradient method can be successfully applied to prepare afatinib liposomes that performed higher encapsulation efficiency and smaller particle size. The UV spectrophotometry employed to determine the liposome encapsulation efficiency was easy operation and with high accuracy.
Afatinib ; Capsules ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Drug Compounding ; methods ; Liposomes ; Lung Neoplasms ; drug therapy ; Quinazolines ; administration & dosage ; chemistry ; therapeutic use

Objective:To establish rat diabetes and eye disease models by injection of STZ and explore the therapeutic effect of panaxnotoginseng polysaccharides (PNP) on the diabetes and eye diseases of the model rats,and to clarify their mechanisms.Methods:Seventy SD male rats were randomly divided into blank control (n=10) and model groups (n=60), and the rats in model group were fed with high fat diet. 2 weeks later, the rats in model group were intraperitoneally injected with 35 mg·kg-1 STZ to establish the models.And 3 d later, the rats were treated with fasting and water deprivation for 12 h,the fasting blood glucose (FBG)was tested, and the models were assessed to be successful as the FBG>11.1 mmol·L-1.The rats with hyperglycemia were selected and divided into model, melbine(150 mg·kg-1), and low, middle and high doses (75,150 and 300 mg·kg-1) of PNP groups.After orally administration for 5 and 8 weeks, the FBG levels of rats were recorded.And 8 weeks later, the sugar tolerance, hepatic glycogen levels,serum glutathione(GSH) and nitric oxide(NO) levels of the rats were tested.The rat retinas were removed to analyze the expression levels of vascular endothelial growth factor(VEGF) and inducible nitric oxide synthase(iNOS) by using Q-PCR.The pathological changes of retinas were observed by HE staining method.Results:Compared with model group,the FBG level in middle dose of PNP group was decreased 5 weeks after treatment(P<0.05).Eight weeks later, compared with model group, the levels of FBG, sugar tolerance and hepatic glycogen in different doses of PNP groups were all decreased (P<0.05 or P<0.01).Compared with model group, the level of serum GSH in high dose of PNP group was remarkably increased(P<0.01), and the NO level was significantly decreased (P<0.05 or P<0.01).Compared with model group, the expression levels of VEGF and iNOS in high dose of PNP group were significantly decreased (P<0.05).The results of HE staining showed that the neurodeatrophia of the rats in low and middle doses of PNP groups were improved;and the vascular proliferation and neurodeatrophia of the rats in high dose of PNP group were significantly improved.Conclusion:PNP could decrease the blood sugar, increase the levels of GSH and NO, and up-regulate the gene expression levels of VEGF and iNOS, resulting the treatment of diabetes and its related retinopathy.

Objective:To establish the rat diabetic models by introperitoneal injection of streptozotocin (STZ) to observe the expressions of MMP-2 in retina tissue of the diabetic rats at different periods,and to clarify the effect of MMP-2 in the diabetic retiropathy (DR) of the diabetic rats.Methods:The femal SD rats were divded into normal control group (n=24),4-week model group (n=30),6-week model group (n=30) and 8-week model group (n=30).The rats in model groups were intraperitoneally injected with STZ for consecutive 5 d.The rats in normal control group were injected with sodium citrate solution at the same volume.The body weights and blood glucose levels of the rats in each group were measured at 4,6,and 8 weeks.The retina of each rat was removed;RT-PCR and Western blotting methods were used to detect the expression levels of MMP-2 mRNA and protein,and the immunohistochemistry was conducted to observe the morphology.Results:The body weights and blood glucose levels of the rats in each group had no differences (P＞0.05) before modeling.Three rats died in 4-week model group,5 rats died at 6 weeks and 7 rats died at 8 weeks.The body weights of the rats in model group were significantly lower than those in normal control group at the same time (P＜0.05 or P＜0.01),and the fasting blood glucose levels were signifieantly increased (P＜0.05 or P＜0.01).The expression levels of MMP-2 mRNA and protein in the retina tissue of the rats in model group were significantly higher than those in normal control group (P＜0.05).In normal control group,the retinal structure was clear and the cells were arranged in order,and the ganglion cells were arranged in a single layer;in model group,the retinal tissue structure was loose,the number of ganglion cells were significantly reduced,the inner nuclear layer and rod cell layer cell membrane outside were fuzzy.The MMP-2 positive cells were found to be brown yellow granules,especially in the cytoplasm of ganglion cells and vascular endothelial cells.The positive expression levels of MMP-2 in model group were higher than those in normal control group at 4,6,and 8 weeks.Conclusion:MMP-2 can express in the retina tissue of the diabetic rats.

BACKGROUND: Neuroscience and brain science researches have paid attention to the effect of astragalus membranaceus in the treatment of neurologic impairment disease and neural regeneration. Studying astragalus membranaceus effects on neural stem cells are becoming a new research direction. OBJECTIVE: To explore the effects of astragalus injection on biological viability of rat neural stem cells. METHODS: Neural stem cells of Wistar rats were separated and cultured. Immunofluorescence staining was applied to identify the neural stem cells. The purified cells were gained by the second subcultivation in vitro, and then the cells were randomly divided into control group and astragalus injection groups with various concentrations (50, 200, 400 g/L) to culture for 6, 12 and 24 hours. The activity of cells was tested by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, and then the immunohistochemistry was applied to detect the expressions of neuron-specific enolase and glial fibril ary acidic protein in the 50 g/L astragalus injection group after induced for 7 days. RESULTS AND CONCLUSION: The viability of neural stem cells increased significantly after intervention with different concentrations of astragalus injection for 6 hours as compared with the control group (P < 0.05). However, there was no difference in the cel viability after treated with different concentrations of astragalus injection for 24 hours (P > 0.05). Compared with the control group, the cells in the 50 g/L astragalus group differentiated rapidly, and the number of positive cells for neuron-specific enolase was increased significantly (P < 0.05). The neural stem cells proliferation was hastened, and its differentiation was promoted by the interference of astragalus injection.