伤寒沙门菌非编码RNA617的分子鉴定及其对生物膜形成的影响研究

本研究旨在探讨伤寒沙门菌(Salmonella enterica serovar Typhi, S. Typhi)中非编码RNA617(non-coding RNA617,ncRNA617)的分子特性,并研究其对生物膜形成的影响及作用机制。采用Northern blot方法检测ncRNA617的表达,通过cDNA 5’末端快速扩增技术(5’-rapid amplification of cDNA end,5’RACE)和逆转录-聚合酶链式反应(reverse transcriotion-polymerase chain reaction,3’RT-PCR)实验分析ncRNA617可能的转录起始位点和终止位点;构建ncRNA617缺陷菌株、回补菌株和过表达菌株等相关菌株,通过生物膜形成实验,观察ncRNA617对伤寒沙门菌生物膜形成的影响,并用实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qPCR)分析生物膜形成相关基因表达水平的变化,综合运用生物信息学方法预测ncRNA617和差异基因的结合区域,初步分析ncRNA617发挥调控作用的机制。结果显示,伤寒沙门菌确有ncRNA617的表达,长度约300 nt,其转录起始位点位于mig-14终止密码子下游967 nt处,终止位点位于t2681起始密码子上游 2 378～2 560 nt处。与野生对照菌株相比,ncRNA617缺陷菌株生物膜形成能力增强(P<0.05),回补菌株的生物膜形成能力恢复至野生菌株水平,过表达菌株的生物膜形成能力有所下降(P<0.05)。qPCR结果表明,ncRNA617可负向调控多个生物膜形成相关基因的转录表达水平(P<0.05)。经生物信息学方法预测发现,ncRNA617与差异基因有不同的结合区域。本研究结果提示,ncRNA617在伤寒沙门菌中存在,其长度约270～452 nt。ncRNA617可能通过靶向结合生物膜形成相关基因下调基因表达,从而负向调控伤寒沙门菌生物膜的生成。     

肠道菌群对耐碳青霉烯类肠杆菌科细菌定殖抗性的研究进展

耐碳青霉烯类肠杆菌科细菌(carbapenem-resistant Enterobacteriaceae,CRE)在肠腔中定殖通常先于或并存于CRE的感染.正常情况下,定殖的CRE、肠道菌群和宿主相互作用,处于稳定平衡的状态,当肠道菌群出现失调时,肠道正常菌群失去对定殖CRE的抵抗力,增加CRE感染的风险.大量研究表明...     

The functional roles,cross-talk and clinical implications of m6A modification and circRNA in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. HCC has high rates of death and recurrence, as well as very low survival rates. N6-methyladenosine (m6A) is the most abundant modification in eukaryotic RNAs, and circRNAs are a class of circular noncoding RNAs that are generated by back-splicing and they modulate multiple functions in a variety of cellular processes. Although the carcinogenesis of HCC is complex, emerging evidence has indicated that m6A modification and circRNA play vital roles in HCC development and progression. However, the underlying mechanisms governing HCC, their cross-talk, and clinical implications have not been fully elucidated. Therefore, in this paper, we elucidated the biological functions and molecular mechanisms of m6A modification in the carcinogenesis of HCC by illustrating three different regulatory factors ("writer", "eraser", and "reader") of the m6A modification process. Additionally, we dissected the functional roles of circRNAs in various malignant behaviors of HCC, thereby contributing to HCC initiation, progression and relapse. Furthermore, we demonstrated the cross-talk and interplay between m6A modification and circRNA by revealing the effects of the collaboration of circRNA and m6A modification on HCC progression. Finally, we proposed the clinical potential and implications of m6A modifiers and circRNAs as diagnostic biomarkers and therapeutic targets for HCC diagnosis, treatment and prognosis evaluation.     

微核形成与细胞周期关系的初步研究 Ⅲ.丝裂霉素c诱发人淋巴细胞染色体畸变与不同周期阶段的微核形成

为了研究染色体畸变与微核形成的关系,本实验用不同浓度的丝裂霉素C(MMC,0.025—0.4μg/ml),处理人外周血淋巴细胞,观察中期染色体畸变与不同细胞周期形成的微核间的关系。获得如下主要结果:(1)MMC诱发的染色体畸变细胞率(ACF),未经培养的G_0期淋巴细胞的微核细胞率(NC-MNCF)以及培养的淋巴细胞的微核细胞率(C-MNCF),在一定剂量范围内均呈剂量依赖性增加,并可用幂回归方程描述;(2)微核形成与染色体畸变全然无关的NC-MNCF,和C-MNCF一样,与ACF呈良好的正相关;(3)用胞质分裂阻滞(CB)法,检测MMC诱发的CB-MNCF,较C-MNCF无显著提高,MNCF/ACF的比值较小,并随着MMC剂量增加从0.15左右降到0.03。所有上述结果表明,不能简单理解微核形成与染色体畸变间的关系,在分裂的细胞群体中,中期染色体畸变可能仅是微核形成的一种来源。     

The NSR1 gene encodes a protein that specifically binds nuclear localization sequences and has two RNA recognition motifs

We previously identified a protein (p67) in the yeast, Saccharomyces cerevisiae, that specifically recognizes nuclear localization sequences. We report here the partial purification of p67, and the isolation, sequencing, and disruption of the gene (NSR1) encoding this protein. p67 was purified using an affinity column conjugated with a peptide containing the histone H2B nuclear localization sequence from yeast. Using antibodies against p67 we have cloned the gene for this protein. The protein encoded by the NSR1 gene recognizes the wild-type H2B nuclear localization sequence, but does not recognize a mutant H2B sequence that is incompetent for nuclear localization in vivo. Interestingly, the NSR1 protein has two RNA recognition motifs, as well as an acidic NH2 terminus containing a series of serine clusters, and a basic COOH terminus containing arg-gly repeats. We have confirmed the nuclear localization of p67 by immunofluorescence and found that a restricted portion of the nucleus is highlighted. We have also shown that NSR1 (p67) is required for normal cell growth.     

2种调查方法对四川黑竹沟国家级自然保护区3种雉类种群密度调查的比较

种群数量是物种的重要生态学基础资料,合适的密度调查方法是数量估算的基础。2016年4—5月,采用广泛应用于鸡形目Galliformes鸟类种群密度调查的样线法和样点法,调查了四川黑竹沟国家级自然保护区3种鸡形目鸟类(白腹锦鸡Chrysolophus amherstiae、红腹角雉Tragopan temminckii和血雉Ithaginis cruentus)的种群密度。样线法和样点法估算的雄体密度分别是:白腹锦鸡1.20只/km²和(6.31±0.98)只/km²,红腹角雉5.41只/km²和(0.39±0.17)只/km²,血雉3.01只/km²和(5.97±2.70)只/km²。除红腹角雉外,样点法估算的白腹锦鸡、血雉种群密度均大于样线法。建议针对不同鸡形目鸟类采用不同的调查方法,并尽量扩大样本数量,从而提高调查结果的准确性。     

Functional analysis of slow myosin heavy chain 1 and myomesin-3 in sarcomere organization in zebrafish embryonic slow muscles

Myofibrillogenesis, the process of sarcomere formation, requires close interactions of sarcomeric proteins and various components of sarcomere structures. The myosin thick filaments and M-lines are two key components of the sarcomere. It has been suggested that myomesin proteins of M-lines interact with myosin and titin proteins and keep the thick and titin filaments in order. However, the function of myomesin in myofibrillogenesis and sarcomere organization remained largely enigmatic. No knockout or knockdown animal models have been reported to elucidate the role of myomesin in sarcomere organization in vivo. In this study, by using the gene-specific knockdown approach in zebrafish embryos, we carried out a loss-of-function analysis of myomesin-3 and slow myosin heavy chain 1 (smyhc1) expressed specifically in slow muscles. We demonstrated that knockdown of smyhc1 abolished the sarcomeric localization of myomesin-3 in slow muscles. In contrast, loss of myomesin-3 had no effect on the sarcomeric organization of thick and thin filaments as well as M- and Z-line structures. Together, these studies indicate that myosin thick filaments are required for M-line organization and M-line localization of myomesin-3. In contrast, myomesin-3 is dispensable for sarcomere organization in slow muscles.     

Expression of Tumor-Associated Macrophage in Progression of Human Glioma

The aim of this study is to investigate the expression of tumor-associated macrophages (TAMs) M1, M2 phenotypic in human glioma tissues, and to explore the clinical significance and prognostic value of TAMs in glioma patients. A total of 50 glioma samples were obtained from patients diagnosed in our hospital from 2007 to 2010. Clinical follow-up was conducted via return visits and telephone interviews after discharge. Progression free survival (PFS) was calculated based on tumor progression by MRI and CT examination from the primary operation. Overall survival (OS) time was calculated from the initial surgical operation date to end date of follow-up or death. Kaplan–Meier methodology was used to evaluate the survival of patients and log-rank test for comparing differences between groups. The expression levels of CD16 and CD206 were investigated in the 4 μm serial paraffin sections by immunohistochemistry. M1-type macrophages filtrated in all the grades of glioma samples, and the lower expression level was associated with high grade glioma. A negative correlation was found between WHO pathological grades and the expression of M1-type macrophages by Spearman correlation analysis. M2-type macrophages filtrated in all the grades of glioma samples with the higher expression level associated with high grade glioma. A positive correlation was found between WHO pathological grades and the expression of M2-type macrophages by Spearman correlation analysis. The PFS and OS among patients with high levels of M1-type macrophages (CD16+++) were significantly higher than those with less expression. The PFS and OS among patients with high levels of M2-type macrophages (CD206+++) were significantly lower than those with low expression. M1-type macrophages may inhibit the tumor growth and improve the therapeutic outcome of glioma patients. M2 ratios are associated with tumor proliferation and poor prognosis. TAMs phenotypes of glioma samples are the potential biomarkers in assessing the degree of malignancy, tumor invasion, and patient prognosis in clinic.     

菟丝子及其一种伪品的鉴别

通过原植物鉴定、药材性状和粉末显微观察,理化反应及紫外光谱扫描比较,对菟丝子(Cuscuta chinensis)及其一种伪品进行鉴别。结果表明,菟丝子及其伪品在原植物的叶、花和果实、药材质地和气味,种皮栅状组织细胞的构成及其特点、热水浸煮时发生的现象、黄酮反应结果以及紫外光谱扫描显示的最大吸收波长位置,均存在明显差异。菟丝子的伪品系十字花科植物芥菜(原变种)(Brassica juncea var.juncea)的干燥成熟种子,上述实验结果为鉴别菟丝子及其伪品提供了依据。