Context:Duchesnea indica is effective against hepatocellular carcinoma (HCC); however, its underlying mechanism of action remains unclear. Objective: The present study aimed to investigate the potential mechanism of action and effects of D. indica components against HCC. Materials and methods: First, the effects of D. indica against HCC were investigated in vitro and in vivo. For in vitro experiments, HCC cell lines were treated with D. indica solutions at different concentrations (0, 1, 2 mg/mL) and then assessed for cell apoptosis, proliferation, migration, invasion, and angiogenic ability. For in vivo experiments, 24 mice were randomly divided into the following four groups: model group and D. indica low-, medium-, and high-dose groups. Tumor growth and CD34 and Ki67 expression levels were assessed to determine the effects of D. indica on cell proliferation and angiogenic ability. Furthermore, transcriptome sequencing and differential expression analyses were used to identify D. indica-induced differentially expressed genes (DEGs) in HCC cells. Additionally, mass spectrometry was conducted to identify the chemical components of D. indica. Four databases were used to predict the target proteins of these chemical components in HCC. HCC-associated genes were identified from two databases. By intersecting the identified DEGs; target proteins; and HCC-associated genes, key D. indica-regulated HCC-related genes were identified. Subsequently, protein-protein interaction network, network pharmacology, and molecular docking were used to identify the active compounds in D. indica and their likely gene targets. Results:In vitro experiments demonstrated that D. indica induced tumor cell apoptosis and inhibited cell proliferation, migration, invasion, and angiogenic potential. In vivo experiments demonstrated that D. indica inhibited tumor growth in a dose-dependent manner. Bioinformatic analyses identified 49 key D. indica-regulated HCC-related genes, of which FOS, SERPINE1, AKR1C3, and FGF2 were the most significant. Mass spectrometry identified the following five molecules in D. indica with potential anti-HCC activity: 4', 5, 7-trihydroxyflavone; ethyl protocatechuate; 3, 5-dihydroxy-benzoic acid; curculigosaponin A; and curculigine G. Molecular docking validated the interaction between D. indica active compounds and their target proteins in HCC. Conclusions: The present study confirmed the therapeutic effects of D. indica against HCC and identified the key genes and active components that may contribute to its mechanism of action, thereby providing a basis for further research on targeted therapeutics for HCC.

脊髓灰质炎(简称脊灰)是由脊灰病毒感染引起的急性传染病,严重者可瘫痪甚至死亡,全球免疫接种疫苗是消灭脊灰的唯一解决方案[1].几十年来,口服脊灰减毒活疫苗(oral polio vaccine,OPV)一直是免疫接种的支柱,是根除野生型脊灰病毒工作的重要组成部分.OPV能够在胃肠道中复制,诱导肠道黏膜免疫,并在个体随后接触活病毒时减少病毒脱落,有效建立免疫屏障,且具有产能高、价格便宜、使用方便等优点[2].

Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.

Abnormal expression of O-Linked β-N-acetylglucosamine (O-GlcNAc) and β-catenin is a general feature of cancer and contributes to transformed phenotypes. In this study, we identified the interaction between O-GlcNAc and β-catenin, and explored their effects on the progression of liver cancer. Our results demonstrated that upregulation of O-GlcNAc was induced by high glucose, whereas the application of PuGNAc and GlcNAc increased β-catenin protein expression levels, as well as the protein's stability and nuclear accumulation in the liver cancer cell lines HEP-G2 and HuH-7. In addition, overexpression of β-catenin could increase O-GlcNAc expression levels through upregulation of uridine 5'-diphosphate (UDP)-N-acetylglucosamine pyrophosphorylase 1 (UAP1) protein expression, protein stability, and inhibition of its ubiquitination. Moreover, the O-GlcNAcylation of β-catenin promoted the proliferation, colony formation, and repressed the induction of apoptosis in HEP-G2 and HuH-7 cells. Knockdown of β-catenin reduced cell proliferation, colony formation, and tumorigenesis, and promoted cell apoptosis through the downregulation of UAP1 expression. In conclusion, this study revealed that the reciprocal regulation between O-GlcNAcylation and β-catenin facilitated the proliferation of liver cancer.

目的 考察口服Ⅰ型Ⅲ型脊髓灰质炎减毒活疫苗在不同温度和pH环境下的稳定性.方法 将6批疫苗分别于2～8℃放置6个月、22～25℃放置7d、37℃放置7d、-20℃放置27、30、33个月,或反复冻融5次;将疫苗pH调至3.0～10.0.采用细胞培养半数感染量(cell culture infective dose 50％,CCID50)法检测病毒总滴度及分型滴度.结果 口服二价脊髓灰质炎减毒活疫苗在2～8℃保存6个月,22～25℃保存5d,37℃保存2d,反复冻融5次,-20℃保存27个月,pH3.0～10.0环境下,疫苗效力均符合质量标准.结论 疫苗的保存温度及时间对其稳定性均有影响.

目的:疫苗的免疫程序和疫苗使用指导原则是保证疫苗免疫效果的重要环节,也是《疫苗管理法》的重要内容之一.早产儿免疫系统发育不成熟,对于各种疫苗免疫反应低于足月儿.本综述为制定早产儿免疫程序提供参考.方法:本文对在早产儿中使用灭活脊髓灰质炎疫苗、百日咳-白喉-破伤风疫苗、乙型肝炎疫苗、肺炎链球菌疫苗、脑膜炎球菌结合疫苗和流感嗜血杆菌疫苗、轮状病毒疫苗、麻疹、腮腺炎、风疹和水痘疫苗等疫苗的免疫效果、安全性及耐受性研究进展进行了归纳整理.结果:本文对早产儿在接种不同疫苗、采取不同免疫策略时体液免疫和细胞免疫效果的研究进行了归纳总结,结果表明虽然早产儿的免疫功能相对较弱,但参照足月新生儿免疫方案接种疫苗后,大部分早产儿可以具有足够的免疫保护力.结论:本文为疫苗免疫程序的制定提供更全面的理解,为制定疫苗使用指导原则提供参考.

基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)在生物大分子的检测中正得到越来越多的使用.为了对质谱在微生物领域的应用提供参考,本文对质谱的原理进行了概述,并对其在微生物鉴定、分型、耐药性研究等方面的应用进行了总结.目前,该法已经在对革兰阴性菌、革兰阳性菌、酵母以及真菌鉴定方面得到应用;在分型方面,已经发现大肠杆菌、金黄色葡萄球菌等越来越多细菌特征性的生物标签,为微生物质谱分型研究打下基础;在细菌耐药性方面,已经能够通过质谱方法区分耐甲氧西林金黄色葡萄球菌和耐万古霉素粪肠球菌等耐药菌群.该法具有快速、准确、便捷和廉价等优点,为微生物学研究提供了新的手段.

We aim to conduct a meta-analysis of studies on the effect of Aidi injection combined with TACE in the treatment of hepatocellular carcinoma (HCC). China National Knowledge Infrastructure (CNKI), Wanfang Database, Chinese Biomedical Literature Database (CBM), Chinese Science and Technology Periodical Database (VIP), Allied and Complementary Medicine Database (AMED), EMBASE, Web of Science, PubMed, and Cochrane Library databases to October 1, 2017, were searched to collect the studies. The data analysis was performed using RevMan 5.3 software. Totally 20 clinical trials with 774 (the experimental group: 447 cases; the control group: 327 cases) HCC patients were finally included in this meta-analysis. Meta-analysis results showed that Aidi injection combined with TACE can, to some extent, enhance the clinical effect and improve the overall survival. Meanwhile, it can increase HCC patients' quality of life. Additionally, Aidi injection plus TACE can reduce adverse events including leukopenia, gastrointestinal reaction, and liver damage in HCC patients (all P < 0.05). Therefore, Aidi injection plus TACE may significantly enhance the clinical effect, suggesting that the combination of TCM and western medicine is promising. The exact outcome needs rigorously designed performances, multicenter, and large randomized controlled trials.

Objective To develop detection methods for porcine circovirus type 1 (PCV1) — the adventitious agent in rotavirus vaccine contamination incident,in African green monkey kidney cell (Vero cell).Methods Six days after PCV1 infection,virus replication and proliferation in Vero cells were tested with qualitative PCR,quantitative PCR (qPCR),electron microscopy (EM) and in situ hybridization (ISH).Results PCR products amplified from PCV1-infected Vero cell DNA extracts presented specific band in agarose gel electrophoresis.According to qPCR,PCV1 copy number increased to 794.3 times of the original level.The detection limits of qualitative PCR and qPCR were both 102.0 copies/ml.Characteristic viral particles were observed by EM.ISH showed dark positive signal of PCV1 in infected cells.Conclusion The replication and proliferation of PCV1 in infected Vero cells can be detected by the 4 methods used.

目的 分析河南汉族人群HLA基因多态性和超型分布. 方法 采用测序法对111名志愿者的血样进行HLA-A、-B、-C、-DRB1和-DQB1五个座位的基因分型,分别根据5个研究组的HLA超型分型标准进行HLA Ⅰ类和Ⅱ类超型的分析统计. 结果 总共检测出130个HLA等位基因,其中HLA-A座位为21个等位基因,-B座位为41个,-C座位为24个,-DRB1座位为27个,-DQB1座位为17个.依据不同的分型方法,共得到86个超型,其中HLA Ⅰ类51种,Ⅱ类35种.按照5个研究组的分型标准,HLA Ⅰ类的优势超型型别基本相似,其中HLA-A2和-B7的分布差异无统计学意义(P＞0.05).但对于HLAⅡ类分子,超型名称分类标准在不同的研究组均不同. 结论 获得河南汉族人群的超型分布特征,HLA Ⅰ类分子的优势超型为HLA-A3、-A2、-B7、-B44和-B62,但HLAⅡ类分子的超型分类标准尚不统一,有待进一步研究.

BACKGROUND:Tumor necrosis factor-308 (TNF-308) was implied to be associated with the development of non-Hodgkin lymphoma (NHL). The aim of this meta-analysis study was to investigate the association of TNF-308A polymorphism with the susceptibility to, and prognosis of, NHL.METHODS:PubMed, Web of Science, Elsevier, HighWire, Scopus, and Google Scholar were searched up to May 2015. The association of TNF-308 polymorphism with the risk of NHL and prognosis was assessed by odds ratio and hazard ratio, respectively.RESULTS:Overall, TNF-308G>A polymorphism increased the risk of NHL, B-cell lymphomas (BCL), and T-cell lymphomas and decreased the risk of follicular lymphomas. In stratified analysis, increased risk of BCL and diffuse large B-cell lymphomas (DLBCL) were observed in Caucasians and population-based studies, whereas decreased risk of NHL, BCL, and DLBCL were detected in Asians and hospital-based studies. Furthermore, pooled results of 1,192 patients with NHL from five studies suggested that TNF-308A was correlated with shorter progression-free survival and overall survival in patients with NHL, BCL, and DLBCL.CONCLUSION:Current evidence indicated that TNF-308A polymorphism was significantly associated with the risk and prognosis of NHL. Future studies should further confirm these associations in other NHL subtypes and ethnicities.

Objective To compare the relationship between the enzyme‐linked immunosorbent assay(ELISA) reagent and West‐ern blot(WB) confirmation reagent for analyzing the quality lever of human T‐cell lymphotropic virus(HTLV) detection reagent . Methods The WB confirmation reagent was used to detect anti‐HTLV antibody in 156 human serum samples of ELISA prelimina‐ry screening positive .The ELISA cut‐off value(optimal value) was selected by using the two‐graph receiver operating characteristics (TG‐ROC) analytical method .The two‐by‐two table analysis was constructed to analyze the consistency of results detected by the two methods ,moreover the McNemar test was used to evaluate the consistency of detection results .The quality level of HTLV de‐tection reagent was comprehensively evaluated .Results Among 156 serum samples of ELISA preliminary screening positive ,only 40 samples were positive by the WB confirmation ,and other 116 samples were negative .The sensitivity and specificity of ELISA de‐tection reagent obtained by TG‐ROC analysis were 97 .5% and 45 .7% respectively ,the TG‐ROC test also indicated that the detec‐tion results had significant difference between ELISA and WB(P<0 .05) .By adjusting the cut‐off value ,the sensitivity and specific‐ity of ELISA were increased to 88 .8% (parametric method) .In the comparison of the parametric method and the non‐parametric method ,the obtained areas under the curve(AUC) was 0 .923 5(parametric method) ,their results were basically consistent .Conclu‐sion Although above results indicate that the detection results of ELISA reagent are different from those of WB ,but adjusting the cut off value can increase its sensitivity and specificity ,thus increases the reliability of diagnosis result .

Objective To investigate effects of different storage conditions and time on stability of HCV RNA. Methods Totally 22 plasma samples were stored at different conditions as bellow :-20 ℃ for 4 weeks ;4 ℃for 12 days ;room temperature(20-25 ℃) for 7 days ;5 freeze‐thaw cycles. HCV RNA was determined by RT‐PCR to investigate the stability of it. Results The average decline of HCV RNA concentration was less than or equal to 0. 14 lg and ± 95% CI ≤0. 20 lg when samples were stored at -20 ℃ for 4 weeks. The average decline of HCV RNA concentration was less than or equal to 0. 39 lg and ± 95% CI ≥ -0. 50 lg when samples were stored at 4 ℃for 12 days. But ± 95% CI≤0. 24 lg when samples were stored at 4 ℃ less than 12 days. The average decline of HCV RNA concentration was less than or equal to 0. 40 lg and ± 95% CI≥ -0. 53 lg when samples were stored at room temperature for 7 days. But ± 95% CI≤0. 29 lg when samples were stored at room temperature less than 7 days. The average decline of HCV RNA concentration was less than or equal to 0. 10 lg and ± 95% CI≤0. 15 lg when samples were frozen and thawed for 5 times. Conclusion HCV RNA is stable in plasma samples stored at -20 ℃ for 4 weeks, 4 ℃ for 9 days, room temperature for 5 days or after 5 freeze‐thaw cycles. But HCV RNA concentration de‐clined dramatically when samples were stored at 4 ℃ for 12 days and room temperature for 7 days. The storage and transportation should be avoided in these conditions.

目的 研制甲型流感病毒抗原检测试剂盒质控参考品.方法 收集甲型流感病毒阳性样本、乙型流感病毒阳性样本和非流感病毒样本,采用流感病毒抗原检测试剂盒方法对样本进行验证.通过协同标定参考品,确定最低检出限范围,3家单位进行适用性检测.反复冻融观察其稳定性.结果 所用的16株病毒经3家单位联合检测,特异性良好,均为甲型流感病毒,检测结果符合率为100％;反复冻融5次后,稳定性良好.结论 建立了第1套甲型流感病毒抗原检测试剂盒质控参考品及相应的质量标准.

Objective To develop the new national reference panel for testing intestinal virus EV71 IgM antibody. Meth-ods The serum samples and throat swabs from HFMD patients were collected, and the swab samples were used for further virus isolating, identifying and genotyping. The EV71 IgM antibody positive samples were verified by neutralization test and ELISA ( capture method) to establish EV71 IgM antobody positive samples and construct the national reference panel. Re-sults The EV71 virus from swabs were all isolated in 12 patients. These virus resulted in the pathological changes of RD cells. The isolated virus were confirmed as epidemic G4 subtype virus by the nested PCR, sequencing and phylogenetic tree analysis. The double samples from each patient could neutralize the EV71 virus. The antibody titers ranged from 1 ∶ 512 to 1 ∶ 2 048, and the tested results of IgM antibodies against EV71 virus were all positive. The EV71 IgM antibody positive reference was developed using 12 positive serum samples as raw materials, and at the same time, the negative reference for EV71 IgM antibody testing was also developed. New national reference panel for testing intestinal virus EV71 IgM antibody was tested by ELISA in the different laboratories. Conclusions The testing panel was suitable for the evaluation of IgM antibody against EV71 virus. It could be used as the reference of EV71 IgM diagnostic reagents. The sensitivity and accura-cy of the panel was in line with the pharmacopeia requirements of the national reference materials of the P. R. China.

Objective To evaluate five kinds of domestic RT-PCR kits for detection of Influenza A(H1N1)virus.Methods The kits were detected on the national reference panel of Influenza A(H1N1)virus respectively,according to the instructions from kits.Results The detected kits for Influenza A(H1N1)virus meet the national standards,however,there were differences in the minimal detection limit and precision among kits from different manufactures and even some different batches of kit from same manufacture.Conclusion The five kinds of domestic PCR-kits for Influenza A(H1N1)virus are reliable in use and provide the foundation for the production,research and clinical evaluation of this kind of reagents.

Objective To establish the new national reference panel in detection of HCMV IgM antibodies.Methods 5 positive samples and 12 negtive samples of HCMV IgM antibodies were collected,respectively.Identification and verification were carried out by uing six commercial HCMV detection kits registered in China and/or foreign countries.Results The sensitivity and accuracy of the panel are in line with the requirements of national reference material of P.R.China.Conclusions The panel is suitable to be used as the reference in quality control for HCMV IgM diagnostic reagents.

流感病毒流行广泛、种类繁多、变异迅速,是人类最主要的健康威胁之一.在流感预防控制的不同环节,对流感病毒诊断试剂的灵敏度、特异性、简便性和使用成本有不同的要求.患者标本类型和采样质量对流感病毒诊断的准确性有较大影响.本文就当前国内外流感病毒诊断技术的研究现状和发展趋势进行了综述.

EB病毒可以导致多种疾病,其与鼻咽癌的关系已得到确认。近年来发现EB病毒在人体内感染情况对于鼻咽癌的诊断、临床分期、预后判断和监测放疗后转移复发等均具有重要意义。各种EB病毒检测方法应运而生。本文就EB病毒诊断方法进展及其意义作一综述。