Detection of peripheral blood T lymphocyte subpopulations among patients positive for both hepatitis B virus surface antigen and surface antibody / 中国热带医学

@#Abstract: Objective　To investigate the frequency of peripheral blood T cells among patients positive for both hepatitis B virus surface antigen (HBsAg) and surface antibody (HBsAb). Methods　 Thirty six patients with co-existence of HBsAg and HBsAb diagnosed were enrolled as the experimental group, who were admitted by Shanghai tenth people's hospital and Wuxi 9th people's hospital from 2014 to 2020. while 40 patients tested positive for HBsAg and negative for HBsAb served as controls, who were admitted by Wuxi 9th people's hospital. Flow cytometry was used to detect and compare the proportions of peripheral blood CD3+, CD4+ and CD8+ T cells between the experimental and control groups. In addition, the associations of serum HBsAb level with peripheral blood T cell proportions, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were examined among chronic hepatitis B patients with co-existence of HBsAg and HBsAb using Pearson correlation analysis. Results　The median age, gender distribution, mean ALT and AST concentrations, proportion of HBV DNA viral load>103 copies/mL, seroprevalence of HBV E antigen (HBeAg), seroprevalence of HBV E antibody (HBeAb), seroprevalence of HBV core antibody (HBcAb) were comparable between the experimental and control groups, and there were no significant difference in them (P>0.05). There were no significant difference between the experimental and control groups in terms of CD3+ T cell proportion [(71.83±1.50)% vs (72.75±1.47)%; t=0.66, P>0.05], CD4+ T cell proportion [(36.81±1.53)% vs (39.88±1.57)%; t=1.43, P>0.05] and CD8+ T cell proportion [(33.17±2.04)% vs (32.40±1.75)%; t=0.77, P>0.05]. Pearson correlation analysis revealed that the serum HBsAb level did not significantly correlate with peripheral blood CD3+ (r=0.026, P=0.65), CD4+ (r=‒0.08, P=0.16) and CD8+ T cell proportions (r=0.09, P=0.24), CD4+/CD8+ T proportion (r=‒0.005, P=0.35), serum ALT (r=0.04, P=0.56) and AST levels (r=0.002, P=0.69) among chronic hepatitis B patients with co-existence of HBsAg and HBsAb. Conclusions　There are no significant differences between HBsAg+/HBsAb+ and HBsAg+/HBsAb- CHB patients in terms of peripheral blood CD3+, CD4+ and CD8+ T cell proportions.

Phosphorylation of Cofilin-1 Enhances Paclitaxel Resistance of Epithelial Ovarian Cancer Cells by Inhibiting Apoptosis / 生物医学与环境科学（英文）

Objective@#To investigate the molecular mechanism of high phosphorylation levels of cofilin-1 (p-CFL-1) associated with paclitaxel resistance in epithelial ovarian cancer (EOC) cells.@*Methods@#Cells displaying varying levels of p-CFL-1 and CFL-1 were created by plasmid transfection and shRNA interference. Cell inhibition rate indicating paclitaxel efficacy was assessed by Cell Counting Kit-8 (CCK-8) assay. Apoptosis was assessed by flow cytometry and protein levels were detected by western blotting. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the expression levels of phosphokinases and phosphatases of CFL-1. Survival analysis evaluated the correlation between the prognosis of EOC patients and the levels of p-CFL-1 and slingshot-1 (SSH-1).@*Results@#High levels of p-CFL-1 were observed in EOC cells that survived treatment with high doses of paclitaxel. SKOV3 cell mutants with upregulated p-CFL-1 showed impaired paclitaxel efficacy, as well as decreased apoptosis rates and pro-survival patterns of apoptosis-specific protein expression. Cytoplasmic accumulation of p-CFL-1 inhibited paclitaxel-induced mitochondrial apoptosis. SSH-1 silencing mediated CFL-1 phosphorylation in paclitaxel-resistant SKOV3 cells. Clinically, the high level of p-CFL-1 and the low level of SSH-1 in EOC tissues were closely related to chemotherapy resistance and poor prognosis in EOC patients.@*Conclusion@#The SSH-1/p-CFL-1 signaling pathway mediates paclitaxel resistance by apoptosis inhibition in EOC and is expected to be a potential prognostic predictor.

Urodynamic assessment of bladder storage function after radical hysterectomy for cervical cancer / 中华医学杂志(英文版)

BACKGROUND@#After radical hysterectomy for cervical cancer, the most common complication is lower urinary tract symptoms. Post-operatively, bladder capacity can alter bladder function for a prolonged period. This study aimed to identify factors affecting bladder storage function.@*METHODS@#A multicenter, retrospective cohort study was conducted. Information of patients with stages IA2 to IIB cervical cancer with urodynamic study results were retrospectively collected from nine hospitals between June 2013 and June 2018 according to the inclusion criteria. Demographic, surgical, and oncological data were collected. The univariate and multivariate logistic regression was used to identify clinical factors associated with bladder storage function.@*RESULTS@#Two hundred and three patients with cervical cancer had urodynamic testing post-operatively. Ninety-five (46.8%) patients were diagnosed with stress urinary incontinence (SUI). The incidence of low bladder compliance (LBC) was 23.2%. Twenty-seven (13.3%) patients showed detrusor overactivity (DO). Fifty-seven patients (28.1%) presented with a decreased maximum cystometric capacity (DMCC). The probability of composite bladder storage dysfunction was 68.0%. Multivariate analysis confirmed that laparoscopy represents a protective factor for SUI with an odds ratio of 0.498 (P = 0.034). Patients who underwent a nerve-sparing procedure were less odds to experience SUI (P = 0.014). A significant positive correlation between LBC and DO was observed (P < 0.001). A greater length of the resected vagina and chemoradiotherapy were common risk factors for LBC and DO, while radiotherapy exerted a stronger effect than chemotherapy. Additionally, patients who received chemoradiotherapy frequently developed a DMCC. The follow-up time was not correlated with bladder storage function.@*CONCLUSION@#A nerve-sparing procedure without longer resected vagina is recommended for protecting the bladder storage function.

TGF-β2 enhances invasion ability of glioma stem cell through matrix metalloproteinase pathway / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To study the effect and possible mechanism of TGF-β2 on the invasion of glioma stem cells (GSCs). Methods: Tumor tissues of 8 patients with glioblastoma multiforme, who underwent resection at Department of Neurosurgery of the FirstAffiliated Hospital of China Medical University duringApril 2016 toApril 2017, were collected. The primary culture of glioma cells were conducted with trypsin digestion. Partial primary glioma cells were seeded into serum-free DMEM/F12 culture medium containing EGF, bFGF and B27 to obtain suspension of tumor spheres. Immunoflurenscent staining and differentiation assay were used to detect whether the tumor spheres were GSCs. TGF-β2 secretion ability of GSCs was determined by ELISAassay.After transfection of TGF-β2 siRNA, the invasion ability of glioma stem cells was determined by Transwell assay. Western blotting was used to examine the effect of TGF-β2 on expression of matrix metalloproteinases (MMP) in glioma stem cells. Results: The suspended tumor spheres were proved to be GSCs by immunofluorescent staining and differentiation assay; the tumor spheres expressed the marker of GSCs（CD133）and had the ability to multi-differentiate (glia and neuronal cells). Compared with the primary glioma cells, Glioma stem cells exerted significantly improved TGF-β2 secretion ability ([74.13±3.63] vs [46.13±2.61] pg/ml, P<0.05); and TGF-β2 silencing significantly reduced the invasion ability of glioma stem cells ([105.71±8.69] vs [63.67±5.93], P<0.05) and inhibited MMP-2 and MMP-9 expressions. Conclusion: TGF-β2 can promote the invasiveness of glioma stem cells through MMP-2 and MMP-9 pathway.

Expression of zinc transporter 1 gene in brain glioma tissues and its effects on proliferation, migration and invasion of glioma U87 cell line / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To detect the expression of zinc transporter 1 (ZnT1) gene in glioma tissue, and to explore its effect on the proliferation, migration and invasion of U87 cells. Methods: From October 2015 to January 2017, 20 patients with glioma, who received no chemoradiotherapy before operation, were collected from Department of Neurosurgery, the First Affiliated Hospital of China Medical University. The protein and mRNA content of ZnT1 in glioma tissues and adjacent tissues were detected by Western blotting and Realtime PCR, respectively. ZnT1 and si-ZnT1 plasmids were transfected into glioma U87 cell line respectively to construct ZnT1 over-expression U87 cell line and ZnT1 knockdown U87 cell line. The effects of ZnT1 on proliferation, migration and invasion of U87 cells were detected by MTT and transwell assay. Results: Both mRNA and protein expressions of ZnT1 in glioma tissues was significantly higher than those in adjacent tissues (all P<0.05). U87 cell lines with ZnT1 over-expression and knockdown were successfully constructed. Compared with the control group and empty plasmid control group, the proliferation (0.54±0.01 vs 0.45±0.04, 0.43±0.03, P<0.01), invasion and migration (all P<0.05) of U87 cells with ZnT1 over-expression were significantly increased at 12 h after transfection; however, the proliferation (0.37±0.03 vs 0.45±0.01, 0.44±0.03, P<0.01), invasion and migration (all P<0.05) of U87 cells with ZnT1 knockdown were decreased significantly. Conclusion: ZnT1 was highly expressed in glioma tissues, and promoted the proliferation, migration and invasion of glioma U87 cells.

Epithelial-interstitial transformation and invasion of glioblastoma multiforme / 中国肿瘤生物治疗杂志

@#[Abstract] Glioblastoma multiforme (GBM) is the most common malignant tumor of the central nervous system. Despite advances in traditional treatment strategies that combine surgery with radiotherapy and chemotherapy, GBM remains one of the most lethal diseases with dismal prognosis. Epithelial-interstitial transformation (EMT) is an important biological process for the invasion and migration of malignant tumors derived from epithelial cells, which is closely related to the pathological behaviors of GBM including invasion, migration, resistance to chemotherapy and radiotherapy. This review will introduce the concept of EMT and its pathophysiological process, especially the latest findings related to the GBM biology. Besides, gene regulation and signaling pathways of EMT (such as matrix metalloproteinases [MMP], TGF-β, transcription factors Snail and Twist etc.) participated in GBM are also introduced, thereby providing a new insight into the fundamental researches and clinical treatments of GBM.

Research progress on the role ofARL2 in the development of tumo / 中国肿瘤生物治疗杂志

@#ADP-核糖基化样因子2（ADP ribosylation factor-like protein 2，ARL2）是一种小型GTP结合蛋白，隶属于RAS超家族 中的ARF家族，广泛存在于真核细胞中，在分子结构上高度保守。ARL2参与调节微管动力学，维持细胞形态和极性；调控线粒 体功能，包括线粒体形态、运动和线粒体融合等。多种肿瘤中存在ARL2表达异常，并且改变肿瘤细胞中ARL2表达会影响肿瘤 细胞的形态、增殖和侵袭能力，影响肿瘤细胞对化疗药物的敏感性、细胞周期分布，甚至诱导细胞凋亡。本文拟对ARL2的目前研 究现状及其在肿瘤中的研究进展作一综述。

Specific microRNA expression profiles of lung adenocarcinoma in Xuanwei region and bioinformatic analysis for predicting their target genes and related signaling pathways / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify differentially expressed microRNAs (miRNAs) related to lung adenocarcinoma in Xuanwei region and predict their target genes and related signaling pathways based on bioinformatic analysis.</p><p><b>METHODS</b>High-throughput microarray assay was performed to detect miRNA expression profiles in 34 paired human lung adenocarcinoma and adjacent normal tissues (including 24 cases in Xuanwei region and 10 in other regions). Gene ontology and KEGG pathway analyses were used to predict the target genes and the regulatory signaling pathways.</p><p><b>RESULTS</b>Thirty-four miRNAs were differentially expressed in lung adenocarcinoma tissues in cases in Xuanwei region as compared with cases in other regions, including 23 upregulated and 11 downregulated miRNAs. The predicted target genes included GF, RTK, SOS, IRS1, BCAP, CYTOKINSR, ECM, ITGB, FAK and Gbeta;Y involving the PI3K/Alt, WNT and MAPK pathways.</p><p><b>CONCLUSION</b>The specific microRNA expression profiles of lung adenocarcinoma in cases found in Xuanwei region allow for a better understanding of the pathogenesis of lung adenocarcinoma in Xuanwei. The predicted target genes may involve the PI3K/Alt, WNT and MAPK pathways.</p>

The Relationship between HIF-2α and VEGF with Radiographic Severity in the Primary Osteoarthritic Knee

PURPOSE: The aim of this study was to determine the relationship of hypoxia-inducible factor-2 (HIF-2α) and vascular endothelial growth factor (VEGF) with radiographic severity in primary osteoarthritis (OA) of the knee. Expression of these two factors in cartilage samples from OA knee joints was examined at mRNA and protein levels. MATERIALS AND METHODS: Knee joints were examined using plain radiographs, and OA severity was assessed using the Kellgren and Lawrence (KL) grading system. Specimens were collected from 29 patients (31 knees) who underwent total knee replacement because of severe medial OA of the knee (KL grades 3 and 4), 16 patients who underwent knee arthroscopy (KL grade 2), and 5 patients with traumatic knees (KL grade 0). HIF-2α and VEGF expression was quantified by real-time polymerase chain reaction and western blotting. RESULTS: Cartilage degeneration correlated with the radiographic severity grade. OA severity, determined using the Mankin scale, correlated positively with the KL grade (r=0.8790, p<0.01), and HIF-2α and VEGF levels with the radiographic severity of knee OA (r=0.7001, p<0.05; r=0.6647, p<0.05). CONCLUSION: In OA cartilage, HIF-2α and VEGF mRNA and protein levels were significantly and positively correlated. The expression of both factors correlated positively with the KL grade. HIF-2α and VEGF, therefore, may serve as biochemical markers as well as potential therapeutic targets in knee OA.

Chemical investigation of triterpenoids from Dichrocephala benthamii / 中国中药杂志

The triterpenoids of Dichrocephala benthamii were investigated by means of silica gel, Sephadex LH-20 and semi-preparative HPLC. Nine triterpenoids were isolated from D. benthamii. By analysis of the EI-MS, NMR spectra and comparison to the data reported in literatures, the structures of these compounds were determined as β-amyrin formiate (1), β-amyrin acetate (2), β-amyrenol (3), β-amyrone (4), 3β-hydroxy-olean-11, 13 (18)-diene (5) , Δ12-oleanene (6) , friedelin (7), dammaradienyl acetate (8), epi-friedeband (9), respectively. Compounds 1-8 were isolated for the first time form this genus, compound 9 was isolated for the first time from this plant, whereas β-amyrin formiate (1) was a new natural product.

Preliminary molecular analysis of bacterial composition in periapical lesions with primary endodontic infections of deciduous teeth / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The bacterial composition of periapical lesions in deciduous teeth has not been well documented. This study was designed to explore the bacterial compositions, especially the dominant bacteria in periapical lesions using 16S rRNA sequencing.</p><p><b>METHODS</b>Tissue samples were collected from 11 periapical lesions in deciduous teeth with primary endodontic infections. DNA was extracted from each sample and analyzed using 16S rRNA cloning and sequencing for the identification of bacteria.</p><p><b>RESULTS</b>All DNA samples were positive for 16S rRNA gene PCR. One hundred and fifty-one phylotypes from 810 clones were identified to eight phyla, and each sample contained an average of 25.9 phylotypes. In addition, 59 phylotypes were detected in more than two samples, and Fusobacterium (F.) nucleatum (8/11), Dialister (D.) invisus (8/11), Campylobacter (C.) gracilis (7/11), Escherichia (E.) coli DH1 (6/11), Aggregatibacter (A.) segnis (6/11), and Streptococcus (S.) mitis (6/11) were the most prevalent species. Furthermore, 45 as-yet-uncultivated phylotypes were also identified.</p><p><b>CONCLUSIONS</b>Chronic periapical lesions in deciduous teeth contained polymicrobial infections. F. nucleatum, D. invisus, C. gracilis, E. coli DH1, A. segnis, and S. mitis were the most prevalent species detected by 16S rRNA sequencing.</p>

Dynamic investigation on the co-infection status of two pathogens in ticks from tourist point in Heilongjiang province / 中华流行病学杂志

Objective To monitor the co-infection status of Borrelia burgdorferi sensu lato (R.b.s.1) and spotted fever group Rickettsia (SFGR) in tourist areas of Heilongjiang province.Methods Polymerase chain reaction (PCR) was used to detect the 5S-23S rRNA intergenic spacer of B.b.s.1 and ompA of SFGR in ticks,dynamically collected from tourist areas of Heilongjiang province in 2010.Amplification products from positive ticks were sequenced,and phylogenetic analysis was conducted by Mega 5.0 software package.Results 849 ticks were collected from two tourist points,with the dominant ticks in Tiger Mountain and Jingpo Lake were Ixodes persulcatus and Haemaphysalis concinna.Regarding the Ixodes persulcatus from Tiger Mountain,the infection rates of B.b.s.1 and SFGR were 26.15％ and 10.05％.The infection rate of SFGR was 13.33％ in Haemaphysalis concinna and the B.b.s.1 was tndiscovered in the same ticks from Jingpo Lake.However,the co-infection could only be detected in Ixodes persulcatus of both tourist areas.Surveillance data showed that the major ticks were more likely to be appeared in July at Tiger Mountain and in June at Jingpo Lake.Data from the sequence analysis on B.b.s.1 showed that the B.b.s.1 in tourist areas could be classified into three different genotypes,other than B.garinii and B.afzelii.We first detected B.valaisiana-like group genotype in northeast of China.Results from the sequence analysis of SFGR positive products showed that the two DNA sequences of newly detected agents were completely the same as Rickettsia sp.HL-93 which was detected in Hulin and Rickettsia sp.H820 found in northeast,China.Conclusion The co-infection of B.b.s.1 and SFGR was detected in ticks from the tourist areas of Heilongjiang province,and data from the sequencing of specific fragment showed that various kinds of genotypes existed in this area.However; the rates of co-infectionitis-different according to environment,time and population that contributed to the kinds of and the index of ticks existed in the surveys points,also the infection rate of the ticks was studied.

Sequence analysis of hemin-binding peptide derived from recombinant hemagglutinin-2 of Porphyromonas gingivalis / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To determine the sequence of the active peptide derived from recombinant hemagglutinin (rHA-2) of Porphyromonas gingivalis (Pg).</p><p><b>METHODS</b>The HA-2 gene from PgATCC33277 was cloned, expressed in Escherichia coli (Ec) BL21 (DE3), and purified. The purified recombinant protein was evaluated for its ability to bind hemin-linked agarose. The active peptide was subjected to endoproteinase-mediated sequence analysis.</p><p><b>RESULTS</b>The protein expressed in Ec BL21 (DE3) was identified as PgHA-2 by plasmid sequence analysis, Western blotting, and mass spectrometry. The recombinant protein was confirmed functional by its ability to bind hemin. The sequence of the active peptide of rHA-2 was determined to be DHYAVMISKTGTNAG.</p><p><b>CONCLUSIONS</b>The availability of sequence of the active peptide of rHA-2 provides a foundation for the development of immunoprophylactic and therapeutic agents against this human pathogen.</p>

Construction of the hemagglutinin-2-deficient mutant of Porphyromonas gingivalis / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To construct and identify the Porphyromonas gingivalis(Pg)ATCC33277 hemagglutinin-2(HA-2)-deficient mutant.</p><p><b>METHODS</b>The genomic DNA of Pg was isolated from PgATCC33277. The up/down stream genes of HA-2-HA(u), HA(l) were amplified by PCR, and inserted into pSY118 separately which contains a 2.1 kb antibiotic resistance ermF-ermAM cassette. The resultant recombinant plasmid-pSY118-HA was linearized as the gene targating fragment HA-ermF-ermAM and used in the electroporation of PgATCC33277. The Pg HA-2-deficient mutant was screened by allelic exchange. The test of aggregation of red blood cells was used to investigate the function change between PgHA2-deficient mutant and the wild type of PgATCC33277.</p><p><b>RESULTS</b>The PgHA-2-deficient mutant was identified by PCR. The ability of Pg HA-2-deficient mutant to aggregate red blood cell was significantly decreased compared with the wild type.</p><p><b>CONCLUSIONS</b>HA-2-deficient mutant of Pg ATCC33277 was constructed successfully, which lays a foundation for further study of its biological function.</p>

Effects of Intravenous Immunoglobulin on Apoptosis and Necrosis of Myocytes in Viral Myocarditis / 实用儿科临床杂志

Objective To investigate the effects of intravenous immunoglobulin(IVIG) on apoptosis and necrosis of myocytes in mice with viral myocarditis.Methods Three hundreds and twenty Balb/c mice were randomly divided into 8 groups.Different courses of IVIG were given in varying time after virus inoculation,Chinese medicine Huangqi given in control group.The virus titer in myocardium、percentage of apoptosis and necrosis of myocytes were detected, myocardial histopathologic scores were counted.Results In every IVIG treatment group,the above 3 items were all significantly lower than that in virus control group and Huangqi group,as IVIG early long course group had the best effect.Conclusion IVIG may reduce the percentage of apoptosis and necrosis of myocytes and virus titer in myocardium in mice with viral myocarditis,the effects are better than that of Huangqi.

Expression of vascular endothelial growth factor in synovium of rats with adjuvant arthritis / 实用儿科临床杂志

Objective To investigate the expression of vascular endothelial growth factor(VEGF)in synovium of rats with adjuvant arthritis and the relationship between the histopathologic score and the expression of VEGF.Methods Adjuvant arthritis was established in Wistar rats by inoculating complete Freund's adjuvant(CFA). We calculated the arthropathologic score and the expression of VEGF mRNA and protein at different stages after CFA inoculation.Results In model group the arthropathologic score and expression of VEGF protein in synovium increased significantly all the time (P