Objective To observe the impacts of the method of fortifying the spleen and supplementing Qi on the cellular immunity of chronic hepatitis B virus(HBV)carriers.Methods Asymptomatic HBV carriers group(ASC group,n=60)with HBV DNA positive were randomly divided into Fortifying the Spleen and Supplementing Qi decoction group(A group,n=30)and conventional treatment group(B group,n=30),the peripheral blood T lymphocyte subsets were detected with flow cytometey and compared with those of the healthy people(The normal control group,n=18).All the patients were treated,and 24 weeks after treatment the indexes were measured again.Results As compared with those of normal controls,the numbers of CD4+ T cells and CD8+ T cells were decreased significantly in HBV carriers(P＜0.01);There was a statistical significance between the values before and complete the treatment with the decoction of fortifying the spleen and supplementing Qi(P＜0.05 or P＜0.01),and compared with conventional treatment group,there Was a statistical significance(P＜0.05).Conclusion T-cell subset function was disturbed in the HBV carriers.The method of fortifying the spleen and supplementing Qi could improve the cellular immunity of carriers with chronic hepatitis B.

The treatment of lower extremity chronic refractory ulcers requires a long time but with poor prognosis. Thus, many patients end up with amputations. The treatment of lower extremity chronic and recalcitrant ulcers and limb salvage has been a challenge worldwide. The Ilizarov technique based on the law of "tension-stress" brings a new hope in treating lower limb chronic and recalcitrant ulcers. The Ilizarov technique and distraction osteogenesis not only induce bone formation but also lead to angiogenesis and improved microcirculation. The Ilizarov technique consists of longitudinal distraction of long bone and tibia trans-verse transport (TTT) (proximal tibial corticotomy followed by transverse distraction). These two techniques have their own advantages and disadvantages with different indications in clinical application. Longitudinal distraction of long bone is mainly used for bone formation, such as large bone defects, osteonecrosis or bone infection (with or without soft tissue loss or ulcers). Because of only a partial osteotomy in TTT, the trauma is minor and their effects on limb instability are limited. Moreover, the procedure is simple with only a few minor complications. Thus, it is ideal in treating lower limb ischemic ulcers, such as diabetic foot ulcers, thromboangiitis obliterans (Buerger's disease), ulcers caused by atherosclerotic occlusion, arterial or venous ulcers, and trauma wounds. Several studies reported that TTT achieved high healing and limb salvage rates in treating severe diabetic foot ulcer. However, TTT could achieve lower recurrent rate. Thus, it is the most successful application in treatment of chronic ulcers. TTT also improves healing and limb salvage in treatment of thromboangiitis obliterans. However, the overall effects are limited than those in treating diabetic foot ulcer. For lower limb atherosclerosis occlusion, TTT induces regeneration of microvessles and consequently leads to ulcer healing. The effects are better than other conventional treatments. A combination therapy with vascularization is emphasized to attain the optimal long-term effects. The effects of TTT on lower limb recalcitrant ulcers still need to be validated in randomized control trial with larger sample size. Further, the mechanism of treatment needs to be explored by pilot studies which could show that this may be related to the formation of pro-angiogenetic factors and a rebalance of the inflammatory microenvironment during TTT.

Objective: To elucidate the mechanism of epigallocatechin gallate (EGCG) mediated anti-HBV effect. Methods: The CCK-8 kit was used to test cell viability in response to EGCG treatment. For HBV DNA replication assay, purified HBV DNA was analyzed by real-time PCR assay. Western blotting was used to confirm HNF4α expression in response to EGCG or siRNA treatment. Results: Our result showed that, EGCG treatment significantly decreasee HBV DNA level both in vivo and in vitro without affecting cell viability. Curiously, we found that EGCG treatment downregulated HNF4α expression. Considering that HNF4α could restrain HBV replication, we knocked down HNF4α in HepG2.2.15 cells and found that the response of HBV replication to EGCG decreased obviously. Conclusions The above result suggest that HNF4α could be an important intracellular factor in EGCG mediated anti-HBV signaling pathway.

Objective:To compare the diagnostic accuracy between multiparametric magnetic resonance imaging (mp-MRI) and biparametric magnetic resonance imaging (bp-MRI) in muscle-invasive bladder cancer (MIBC).Methods:The clinical data of 195 patients with bladder cancer at the First Affiliated Hospital of Nanjing Medical University from July 2020 to June 2022, were retrospectively reviewed. There were 160 males and 35 females, with the median age of 68(61, 76)years old. Mp-MRI was performed on each patient within 6 weeks before transurethral resection of bladder tumor or radical cystectomy. Each patients’ images were divided into two sets. Set 1 (bp-MRI) included the axial, sagittal, coronal T2-weighted images (T2WI), and axial diffusion-weighted images (DWI) or apparent diffusion coefficient maps. Set 2 (mp-MRI) included Set 1 images in addition to dynamic contrast-enhanced images. All images were independently reviewed and evaluated by two radiologists. Mp-MRI was evaluated according to the Vesical Imaging-Reporting and Data System (VI-RADS)guideline, and bp-MRI was evaluated according to two types of criteria. Bp-MRI (Criterion A): VI-RADS scoring is determined 2 when T2WI 3-point with DWI 2-point. Bp-MRI (Criterion B): VI-RADS scoring is determined 3 when T2WI 3-point with DWI 2-point. VI-RADS scoring ≥ 3 or ≥ 4 was used as the cut-off value to predict MIBC. The sensitivity, specificity, positive predictive value, and negative predictive value of mp-MRI, bp-MRI (Criterion A), and bp-MRI (Criterion B) were calculated, as well as receiver operating characteristic curves and the areas under the curve (AUC).Results:Of 195 patients, 135 patients (69.2%) were pathologically confirmed as NMIBC and 60 patients (30.8%) were MIBC. When the VI-RADS cut-off value was ≥ 3, the sensitivity of mp-MRI, bp-MRI (Criterion A), and bp-MRI (Criterion B) were identical, all at 88.3% (53/60). The specificity of bp-MRI (Criterion A), bp-MRI (Criterion B), and mp-MRI were 88.9% (120/135), 73.3% (99/13), and 86.7% (117/135), respectively. When the VI-RADS cut-off value was ≥ 4, both bp-MRI (Criterion A) and bp-MRI (Criterion B) were classified as the same criterion. The sensitivity of bp-MRI and mp-MRI were 70.0% (42/60) and 75.0% (45/60), respectively. The specificity of bp-MRI and mp-MRI were identical, at 95.6% (129/135). The AUC for bp-MRI (Criterion A), bp-MRI (Criterion B), and mp-MRI were 0.927 (95% CI 0.881-0.959), 0.904 (95% CI 0.853-0.941), and 0.927 (95% CI 0.881-0.959), respectively. The AUC for bp-MRI (Criterion A) and mp-MRI were significantly higher than that of bp-MRI (Criterion B) ( P<0.001). There was no significant difference in AUC between bp-MRI (Criterion A) and mp-MRI ( P=0.939). Conclusions:Bp-MRI (Criterion A), VI-RADS scoring is determined 2 when T2WI 3-point with DWI 2-point, shows comparable diagnostic accuracy in predicting MIBC with mp-MRI. Compared to bp-MRI (Criterion B), the corresponding situation when VI-RADS scoring is determined 3, bp-MRI (Criterion A) may have better diagnostic accuracy than bp-MRI (Criterion B) in predicting MIBC.

Objective:In this study, the collected mosquito samples were subjected to viral isolation to identify the species and branch characteristics of arboviruses in five regions of Shanxi Province.Methods:Eight arboviruses in mosquito samples collected from July to September 2020 were detected by real-time fluorescent quantitative PCR, and virus isolation was carried out through cell culture. Virus isolates were identified and analyzed by molecular biology and bioinformatics method.Results:We detected 1 batch of positive samples of Japanese encephalitis virus, 2 batches of positive samples of Culex flavivirus and 8 batches of positive samples of Nam Dinh virus among 121 batches of mosquito samples. Seven virus isolates were isolated, numbered: SX-YJ-Cxp-4、SX-YJ-Ars-2、SX-YJ-Cxp-1、SX-LY-Cxp-10、SX-GP-Ars-5、SX-GP-Cxp-2、SX-GP-Cxp-4, all of which were identified as Nam Dinh virus, and the whole genome sequencing was performed on one of them, and the result showed that Shanxi Nam Dinh virus isolate and Yunnan Nam Dinh virus isolate belonged to the same evolutionary branch.Conclusions:Nam Dinh virus was isolated and identified on the specimen from Shanxi province for the first time.

Objective:To determine the evaluation indexes of national standard strains of genotypes 1, 3 and 5 of Japanese encephalitis virus (JEV) and evaluate the national standard JEV strains.Methods:According to the national standard strain evaluation technical standards of pathogenic microbial bacteria (virus) species, based on the application of Japanese encephalitis virus research, and according to the morphological characteristics, biological characteristics, molecular biological characteristics and other research data to identify the characteristics of G1, G3 and G5 genotypes of JEV.Results:Spherical virus particles with a diameter of about 60 nm were visible under electron microscope of the three Japanese encephalitis virus strains. The cytopathic effect was mainly characterized by cell shrinkage and exfoliation in BHK-21 and Vero cell lines, cell fusion and exfoliation were shown after infection with C6/36 cell line; the virus titer was 10 5-10 7 PFU/ml, and the plaque size was different by genotype. The median lethal dose of intrabitoneal challenge in G1, G3 and G5 JEV in three weeks-old mice was 50.51 PFU, 6.98 PFU, and 8.13 PFU, and the median lethal dose of intracranial challenge in five weeks mice was 3 PFU, 0.3 PFU, 1.35 PFU. The whole genome length of G1, G3 and G5 JEV was 10 967 bp, 10 976 bp and 10 983 bp, respectively. Conclusions:Three genotypic national standard strains of JE V were identified and evaluated by electron microscopy, cell, animal and genome laboratory indexes, which provided reference for the identification and evaluation of other national standard strains of JEV.

Objective:To establish a rapid method for the detection of varicella-zoster virus (VZV) by recombinase-aid amplification (RAA).Methods:The whole genome sequences of VZV were downloaded from the global shared database for comparison and analysis. Specific primers and probe were designed for the four conserved genes respectively and the optimal combination was selected. The optimal reaction system was selected through the concentration gradient of primers and probes, and a fluorescence RAA detection method was established. The sensitivity of the method was evaluated with VZV positive plasmid standard and clinical samples with gradient dilution, the repeatability of the method was evaluated with the lowest detectable limit concentration of positive plasmid standard, and the specificity of other viral nucleic acid method was evaluated. At the same time, this method and quantitative real-time PCR (qPCR) were used to detect clinical samples and the result were compared.Results:The optimal combination of primer pair F2/R2 and probe P2 targeting open reading frame (ORF) 28 gene was selected. Considering the cost factor, the optimal primer concentration was set at 500 nmol/L and the optimal probe concentration was 280 nmol/L. The minimum detection limit was 10 1 copies/μL, and the minimum clinical positive samples with a Ct value of 36.027 could be detected, and the result of repeated experiments were consistent. The method has no cross-reaction with other viral nucleic acids. The detection rate of clinical positive samples was 93.33%, which was almost identical to that of qPCR. Conclusions:This method is simple to operate with high sensitivity, strong specificity, low requirements for experimental conditions, visual detection result, and can detect VZV nucleic acid in samples within 20 minutes, which is a rapid VZV detection method that can be considered for clinical use for detection.

Objective Genotypes(G)1,3,and 5 of the Japanese encephalitis virus(JEV)have been isolated in China,but the dominant genotype circulating in Chinese coastal areas remains unknown.We searched for G5 JEV-infected cases and attempted to elucidate which JEV genotype was most closely related to human Japanese encephalitis(JE)in the coastal provinces of China. Methods In this study,we collected serum specimens from patients with JE in three coastal provinces of China(Guangdong,Zhejiang,and Shandong)from 2018 to 2020 and conducted JEV cross-neutralization tests against G1,G3,and G5. Results Acute serum specimens from clinically reported JE cases were obtained for laboratory confirmation from hospitals in Shandong(92 patients),Zhejiang(192 patients),and Guangdong(77 patients),China,from 2018 to 2020.Seventy of the 361 serum specimens were laboratory-confirmed to be infected with JEV.Two cases were confirmed to be infected with G1 JEV,32 with G3 JEV,and two with G5 JEV. Conclusion G3 was the primary infection genotype among JE cases with a definite infection genotype,and the infection caused by G5 JEV was confirmed serologically in China.

Objective To compare the efficacy of retroperitoneal robot-assisted laparoscopic partial nephrectomy(RALPN)in the treatment of ventral and dorsal renal tumor.Methods The clinical data of 131 patients with renal tumor who underwent retroperitoneal RALPN at our hospital during Jan.2021 and Feb.2024 were retrospectively analyzed.The patients were divided into the ventral renal tumor group(n=35)and dorsal renal tumor group(n=96)according to preoperative images.Perioperative outcomes and prognosis were compared between the two groups.Results All 131 surgeries were successfully completed without conversion to open surgery or abdominal organ injury.There were no significant differences in warm ischemia time[median(interquartile range,IQR):20(16.75)min vs.22(IQR:15.25)min],operation time[57.0(IQR:29.5)min vs.58.5(IQR:39.75)min],estimated intraoperative blood loss[50(IQR:80)mL vs.50(IQR:80)mL],proportion of patients with intraoperative blood transfusion(0 vs.2.1％),proportion of patients using four arms during operation(42.9％vs.37.5％),postoperative reduction of hemoglobin(Hb)[(-12.9±9.0)g/L vs.(-11.5±9.4)g/L],reduction of estimated glomerular filtration rate(eGFR)[(-3.8±12.4)mL/min vs.(-7.0±13.6)mL/min],postoperative hospital stay[4(IQR:0)d vs.4(IQR:2)d],and proportion of meeting"Trifecta"criteria(94.3％vs.86.5％,P＞0.05).During the median follow-up of 9.8(5.5,24.0)months,no death occurred,and no recurrence or metastasis were observed except that bone metastasis occurred in 1 patient in the dorsal renal tumor group.Conclusion Both ventral and dorsal renal tumors can be treated with retroperitoneal RALPN,and the surgical outcomes of both are comparable.

Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.