上海市630例新型冠状病毒Omicron变异株感染者流行病学特征及影响核酸转阴时间的相关因素

【目的】探讨上海市新型冠状病毒Omicron变异株感染者流行病学特征及影响核酸转阴时间的相关因素。【方法】回顾性分析上海黄浦方舱医院于2022年4月至5月收治的630例新型冠状病毒Omicron变异株感染患者的临床资料,按临床分型标准将其分为无症状/轻型组(n=543)和普通型组(n=87),根据胸部CT影像学诊断结果将患者分为A组(胸部CT平扫未见明显异常者,n=208)、B组(胸部影像呈现多发小斑片影及间质改变者,n=335)、C组(胸部影像呈现多双肺多发磨玻璃影、浸润影者,n=87)。比较各组胸部影像学特征及核酸检测结果,分析影响核酸转阴时间的相关因素。【结果】无症状/轻型组和普通型组不同性别、年龄、核酸基因分布比例,以及N基因Ct值、ORF基因Ct值比较,差异均有统计学意义(P<0.05)。A组、B组、C组不同年龄、核酸基因分布比例,以及N基因Ct值、ORF基因Ct值比较,差异均有统计学意义(P<0.05)。Logistic多因素回归分析显示,年龄、N基因Ct值、ORF基因Ct值为患者肺炎重症化的影响因素(P<0.05)。根据核酸转阴时间将其分为D组(核酸转阴时间<9 d,n=5)、E组(核酸转阴时间≥9 d,n=12)。D组、E组患者的年龄、入院时N基因Ct值、ORF基因Ct值比较,差异有统计学意义(P<0.05)。Logistic多因素回归分析显示,年龄、入院时N基因Ct值、ORF基因Ct值并不是影响患者核酸转阴时间的独立危险因素(均P>0.05)。【结论】新型冠状病毒Omicron感染者流行病学特征:性别、年龄及核酸基因Ct值对于临床分型有影响,而对病毒核酸转阴时间并无明显影响;患者肺部发生炎症改变的程度与年龄及病毒核酸含量相关。

流行性感冒暴发疫情实验室检测与分析

人流行性感冒(流感),是由甲(A)、乙(B)、丙(C)3种流感病毒引起的急性呼吸道传染病。流感病毒属正粘病毒科,流感病毒属,是具有包膜和分节段的单链负股RNA病毒。流感病毒经呼吸道传播,由于其传播速度极快,且极易发生抗原漂移和转变等变异,因此,每年流感都会发生不同规模的暴发和流行,给人类造成不同程度的伤害[1-2]。本文通过对深圳市坪山区某学校流感暴发疫情进行实验室检测和结果分析,旨在通过本次疫情了解流感病毒的型别及变异情况,从而了解流感发病规律,对易感人群开展病原学监测。

2010—2014年桂林市流感病毒核酸检测结果分析

目的分析2010年1月—2014年6月桂林市流感病毒的流行情况,为该地区内流感的科学防控提供依据。方法采集桂林市疾控中心监测的流感样病例（ILI）的咽拭子样本,采用荧光定量RT-PCR方法确定病毒的亚型,并分析各亚型流感病毒的流行情况及其与患者性别、流行时间等因素的关系。结果 2010—2014各年份流感监测的阳性率在12.32%-36.62%,4049例样本的总阳性率为22.8%,阳性样本中各型所占比例为B型52%,季节性H1型1%,季节性H3型34%,新甲型H1型13%。患者中男性发病率为24.42%,女性发病率为20.60%,差异无统计学意义,发病人数的高峰期集中在每年的3—5月。结论 2010—2014年桂林地区的流感疫情主要以乙型流感为主,春季为流感发病的高峰期。对流感样病例进行流感病毒核酸检测并对结果进行数据统计,可以更好的判断该地区流感流行的特点,为流感疫情的防控提供有效参考。

Classification and Identification of Marine Penicillium Species Based on ITS Sequences of rDNA

[ Objective ] The aim of this study is to identify 6 marine fungi species by analyzing ITS nucleotide sequence, which had been primarily identified as penicillium based on morphological characteristics. [ Method ] The ITS regions of these species were cloned by molecule biology method and phylogenetic analyzed using ClustalX1.83 software. [ Result] The ITS regions of these species were sequenced, and phylogenetic analysis between the yield sequences and the ITS sequences assessed in GenBank showed that the 6 strains all belonged to penicillium. [ Condusion] The present study suggests ITS sequence analysis could not be used as an only proof, but it is a very useful supplementary tool for the classification and identification of marine peniciUium combined with morphological characteristics.

核酶对肝癌SMMC-7721细胞端粒酶活性及凋亡的影响

目的:观察针对人端粒酶RNA模板区的核酶对肝癌细胞端粒酶活性和细胞凋亡的影响。方法:利用脂质体Lipofectamine介导,将已构建好的带有端粒酶核酶基因的重组质粒pBBS212Rz及空载质粒pBBS212转染肝癌SMMC-7721细胞,采用TRAP-ELISA法检测端粒酶活性,用倒置相差显微镜及流式细胞仪观察细胞生长和凋亡情况。结果:重组质粒pBBS212Rz转染的肝癌SMMC-7721细胞的端粒酶活性明显下降,细胞生长速度明显变慢,凋亡加速。结论:端粒酶核酶对肝癌细胞端粒酶活性和细胞生长有抑制作用,可望成为肝癌基因治疗的方法。

The fidelity of DNA synthesis by eukaryotic replicative and translesion synthesis polymerases

In their seminal publication describing the structure of the DNA double helix , Watson and Crick wrote what may be one of the greatest understatements in the scientific literature, namely that ＂It has not escaped our notice that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic material.＂ Half a century later, we more fully appreciate what a huge challenge it is to replicate six billion nucleotides with the accuracy needed to stably maintain the human genome over many generations. This challenge is perhaps greater than was realized 50 years ago, because subsequent studies have revealed that the genome can be destabilized not only by environmental stresses that generate a large number and variety of potentially cytotoxic and mutagenic lesions in DNA but also by various sequence motifs of normal DNA that present challenges to replication. Towards a better understanding of the many determinants of genome stability, this chapter reviews the fidelity with which undamaged and damaged DNA is copied, with a focus on the eukaryotic B- and Y-family DNA polymerases, and considers how this fidelity is achieved.

Eukaryotic DNA damage tolerance and translesion synthesis through covalent modifications of PCNA

In addition to well-defined DNA repair pathways, all living organisms have evolved mechanisms to avoid cell death caused by replication fork collapse at a site where replication is blocked due to disruptive covalent modifications of DNA. The term DNA damage tolerance （DDT） has been employed loosely to include a collection of mechanisms by which cells survive replication-blocking lesions with or without associated genomic instability. Recent genetic analyses indicate that DDT in eukaryotes, from yeast to human, consists of two parallel pathways with one being error-free and another highly mutagenic. Interestingly, in budding yeast, these two pathways are mediated by sequential modifications of the proliferating cell nuclear antigen （PCNA） by two ubiquitination complexes Rad6-Rad18 and Mms2-Ubc13-Rad5. Damage-induced monoubiquitination of PCNA by Rad6-Rad18 promotes translesion synthesis （TLS） with increased mutagenesis, while subsequent polyubiquitination of PCNA at the same K164 residue by Mms2-Ubc13-Rad5 promotes error-free lesion bypass. Data obtained from recent studies suggest that the above mechanisms are conserved in higher eukaryotes. In particular, mammals contain multiple specialized TLS polymerases. Defects in one of the TLS polymerases have been linked to genomic instability and cancer.

Na^+/K^+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress

Na^＋/K^＋-ATPases are membrane-associated enzymes responsible for the active transport of Na^＋ and K^＋ ions across cell membranes, generating chemical and electrical gradients. These enzymes＇ α-subunit provides catalytic function, binding and hydrolyzing ATP, and itself becoming phosphorylated during the transport cycle. In this study, Na^＋/K^＋-ATPase α-subunit eDNA was cloned from gill tissue of the swimming crab Portunus trituberculatus by reverse-transcription polymerase chain reaction （RT-PCR） and rapid amplification of eDNA end methods. Analysis of the nucleotide sequence revealed that the eDNA had a full-length of 3 833 base pairs （bp）, with an open reading frame of 3 120 bp, 5＇ untranslated region （UTR） of 317 bp, and 3＇ UTR of 396 bp. The sequence encoded a 1 039 amino acid protein with a predicted molecular weight of 115.57 kDa and with estimated pI of 5.21. It was predicted here to possess all expected features of Na^＋/K^＋-ATPase members, including eight transmembrane domains, putative ATP-binding site, and phosphorylation site. Comparison of arnino acid sequences showed that the P. tritubereulatus α-subunit possessed an overall identity of 75%-99% to that of other organisms. Phylogenetic analysis revealed that this α-subunit was in the same category as those of crustaceans. Quantitative real-time RT-PCR analysis indicated that this α-subunit＇s transcript were most highly expressed in gill and lowest in muscle. RT-PCR analysis also revealed that α-subunit expression in crab gill decreased after 2 and 6 h, but increased after 12, 24, 48, and 72 h. In addition, α-subunit expression in hepatopancreas of crab decreased after 2-72 h. These facts indicated that the crab＇s Na^＋/K^＋-ATPase α-subunit was potentially involved in the observed acute response to low salinity stress.

Meta-analysis gene lists about subtypes of leukemia based on gene expression data

To screen for molecular signatures that are commonly dysregulated in subtypes of a certain cancer, a novel meta-analysis is designed to perform rank score （RS） on lists of genes that are derived from different studies. RS is a promising way to detect signatures across platforms when integrating with one vs. all （OVA） or one vs. one （OVO） schemes of comparison. Among six published microarray expression datasets on acute leukemia, the biological signals hereafter provide stronger clustering support than systematic differences among microarray platforms. Moreover, the pediatric BCR_ABL specific genes can be used to correctly discriminate independent adult BCR ABL cases. The obtained results redound to discover, validate and treat the subtypes from microarray gene expression profiles of cancer, which have been plentifully researched, such as leukemia.

Genetic Analysis of the VP1 Region of Human Enterovirus 71 Strains Isolated in Fuyang,China,During 2008

Enterovirus 71 （EV71） is a common cause of Hand, foot, and mouth disease （HFMD） and may also cause severe neurological diseases, such as encephalitis and poliomyelitis-like paralysis. To examine the genetic diversity of EV71, we determined and analyzed the complete VP1 sequences （891 nueleotides） from nine EV71 strains isolated in Fuyang, China. We found that nine EV71 strains isolated were over 98% homologous at the nucleotide level and 93%-100% homologous tO members of the C4 subgenogroup. At the amino acid level, these Fuyang strains were 99% -100% homologous to one another, 97%-100% homologous to members of the C4 subgenogroup, and the histidine（H） at amino acid position 22 was conserved among the Fuyang strains. The results indicate that Fuyang isolates belong to genotype C4, and an H at position 22 appears to be a marker for the Fuyang strains.

Complete Genome Sequence Analysis of Duck Circovirus Strains from Cherry Valley Duck

To investigate molecular epidemiology of DuCV in Cherry Valley ducks in China, the complete genomes of six DuCV strains, which were detected from Cherry Valley ducks in China between 2007 and 2008, were sequenced. Sequence and phylogenetic analysis were carried out to compare these six strains with another 27 DuCV strains from Mulard duck, Muscovy duck, Pekin ducks and Mule duck. The analysis showed that the six DuCV strains exhibited typical genetic features of the family of DuCV, such as a stem-loop structure, three major open reading frames (Rep, Cap and ORF3), four intergenic repeats and the conserved motifs for rolling circle replication and for the dNTP binding domain located in the Rep protein. Phylogenetic analysis of the nucleotide sequences of the complete genome and Cap gene of these strains together with those that have been previously published demonstrated two distinct DuCV genotypes. The DuCV strains with complete genomes containing 1988 and 1989 nucleotides clustered in genotype A, whereas the strains with complete genomes containing 1991, 1992, 1995 and 1996 nucleotides lay in genotype B. The six DuCV strains from Cherry Valley ducks were divided into the two groups. The results of the study provides some insight into the variation of DuCVs in Cherry Valley ducks.

The involvement of XPC protein in the cisplatin DNA damaging treatment-mediated cellular response

Recognition of DNA damage is a critical step for DNA damage-mediated cellular response. XPC is an important DNA damage recognition protein involved in nucleotide excision repair (NER). We have studied the XPC protein in cisplatin DNA damaging treatment-mediated cellular response. Comparison of the microarray data from both normal and XPCdefective human fibroblasts identified 861 XPC-responsive genes in the cisplatin treatment (with minimum fold change≥1.5).The cell cycle and cell proliferation-related genes are the most affected genes by the XPC defect in the treatment. Many other cellular function genes, especially the DNA repair and signal transduction-related genes, were also affected by the XPC defect in the treatment. To validate the microarray data, the transcription levels of some microarray-identified genes were also determined by an RT-PCR based real time PCR assay. The real time PCR results are consistent with the microarray data for most of the tested genes, indicating the reliability of the microarray data. To further validate the microarray data, the cisplatin treatment-mediated caspase-3 activation was also determined. The Western blot hybridization results indicate that the XPC defect greatly attenuates the cisplatin treatment-mediated Caspase-3 activation. We elucidated the role of p53 protein in the XPC protein DNA damage recognition-mediated signaling process. The XPC defect reduces the cisplatin treatment-mediated p53 response. These results suggest that the XPC protein plays an important role in the cisplatin treatment-mediated cellular response. It may also suggest a possible mechanism of cancer cell drug resistance.

高浓度HBVDNA致核酸检测失效1例分析

目前，国内多数血液中心和部分中心血站相继开展了核酸检测，青岛市中心血站作为卫生部首批试点单位自2010—06—01起全面实施。我们采用的Roche cobass 201system，检测发现高浓度HBVDNA致内标失效1例报告如下。

Single Nucleotide Polymorphism Genotyping of Calpastatin Gene Using the ARMS Compared with the RFLP

Calpastatin is an endogenous inhibitor of calpain which is responsible for the breakdown of myofibrillar proteins, The association of Single Nucleotide Polymorphism （SNP） in the calpastatin gene with meat tenderness is an important topic in meat production. Therefore efficient procedure to investigate the SNP is necessary. The objectives of this study were to detect the SNP of calpastatin gene at domain L marker （G/C transversion） of the Kamphaengsaen beef breed （KPS cattle; n = 26） by the Amplification Refractory Mutation System （ARMS） compared with the Restriction Fragment Length Polymorphism （RFLP） methods and to determine the genotypes of the KPS cattle at that marker. Genomic DNA of calpastatin gene extracted from blood of the KPS cattle was detected with ARMS and RFLP methods. The ARMS system has utilized two primer pairs to amplify the two different alleles of a polymorphism in single PCR reaction to detected single base mutation. In this method, the alleles-specific primers had a mismatch at 3＇ terminal base and a second deliberate mismatch at position -2 from 3＇ terminus. While the RFLP method detected a polymorphism using PCR-base technique follow by RsaI restriction enzyme. Amplification of the ARMS method revealed that the results were not different from the conventional method of RFLP. Analysis of genotypes revealed that the KPS cattle inherited the CC, CG and GG genotypes at domain L marker. These were reliable when verified by nucleotide sequence analysis of PCR products. The animals were genotyped and determined for tenderness phenotype with this marker that predicted variation of an intronic polymorphism at domain L of the calpastatin gene. Therefore, the ARMS method was simple, efficient technique, and suitable for detecting SNP at domain L marker of the calpastatin gene.

DTNBP1 Gene Is Associated with Some Symptom Factors of Schizophrenia in Chinese Han Nationality

Objective To study the association of DTNBP1 gene with some symptom factors of schizophrenia.Methods A total of 285 unrelated schizophrenic individuals were recruited from December 2004 to January 2009 for genetic analysis,and their symptom factors were assessed based on the Positive and Negative Syndrome Scale(PANSS).The quantitative trait test was performed by the UNPHASED program(version 3.0.12) to investigate the association between scored positive and negative symptoms and the single nucleotide polymorphisms(SNPs) in DTNBP1 gene.Results The quantitative trait test showed allelic association of rs909706 with the excitement symptom of schizophrenia(P<0.05,adjusted by 10 000 permutations),while the genotype C/G of rs2619539 with a negative symptom,lack of spontaneity and flow of conversation(P<0.05,adjusted by 10 000 permutations).Conclusion DTNBP1 variations are possibly associated with some symptoms of schizophrenia,which could partly explain the relationship between the susceptibility gene DTNBP1 and that disease.

Confirmed Diagnosis by RT-PCR and Phylogenetic Analysis of Peste des Petits Ruminants Viruses in Tibet, China

This paper reports the confirmed diagnosis by nested RT-PCR of PPR cases in Tibet, China in 2007, and results of phylogenetic analysis. Results showed that the 11 tested samples were PPRV positive by nested RT-PCR, of which 2 samples were genetically close to the X7443 strain (Nigeria 75/1) of lineage I, and 3 samples close to the strain AY560591 (Sungri96) of linage IV with 96.6%、97.3%、97.6% and 98% nucleotide sequence homogeneity respectively, based on partial sequencing of the F gene from 5 samples and complete sequencing of the N/M/F/H genes from one sample. This study suggested that there are at least 2 origins of PPRV in China.

Detection of ＂ Candidatus Phytoplasma asteris＂ in Brussels Sprout and Its Possible Association with Flower Bud Failure in Poland

Severe growth abnormalities including shoot stunting, leaf blade reduction and flower bud failure of Brussels sprout were observed in Poland. The presence of phytoplasma in diseased as well as in healthy looking plants, was demonstrated by nested polymerase chain reaction assay employing phytoplasma universal rRNA primer pairs-P1/P7 followed by R16F2n/R16R2. Products of PCR primed by R 16F2n/R 16R2 primer pair from naturally infected Brussels sprouts were sequenced. Comparison of the obtained 16S rDNAs revealed high nucleotide sequence identity between analyzed phytoplasma isolates （99.8%-100%）. They were also nearly identical with the sequences of other phytoplasmas isolates from sub-group 16SrI-B, and they were classified as members of ＂Candidatus Phytoplasma asteris＂.

The role of advanced glycation end products and their mechanism in DN

Advanced glycation end products(AGEs), which are macromolecular material such as proteins, lipids, and nucleic acids free amino and reducing sugar on the reaction of aldehyde group under the condition of the enzyme, generate the stable compounds. AGEs formation is enhanced in diabetes and is associated with the development of diabetic complications. AGEs, as an important marker of chronic complications of diabetes mellitus, plays an important role in the development and progression of diabetic nephropathy. In the current review, we discuss mechanisms and the role of AGEs in diabetic nephropathy.

MinK gene G112A polymorphisms and atrial fibrillation:a Meta-analysis

Atrial fibrillation (AF) is the most common arrhythmia with multi-factorial pathogenesis. A number of studies of genetic epidemiology have assessed the association of G112A (G38S) single nucleotide polymorphisms (SNPs) in Mink gene with AF in different populations. However, the results are inconsistent and inconclusive. We performed a Meta-analysis of the association between G112A polymorphisms of MinK gene and AF to estimate the magnitude of the gene effect. Six case-control studies with a combined 854 cases and 1079 controls were summarized. Subgroups in different races were separately analyzed. Heterogeneity and publication bias were also explored. When all groups were pooled, the individuals with G allele had an over 40% higher risk of AF compared with individuals with the A allele. The GG genotype (versus AA genotype) was found to be significant association with increased AF risk. The significant associations were also found in both dominant and recessive genetic model. For subgroup analysis, the results were consistent with above, except that the pooled OR for Chinese population was not significant in a recessive genetic model. In conclusion, G112A polymorphisms in Mink gene may have an important effect on the pathogenesis of AF. This warrants further investigation in large multi-center studies with precise design.

Variation of DRB1 Gene in Tibetan Sheep

It reveals that the MHC （major histocompatibility complex） gene product always involved in the control of immune response and disease resistance. Nowadays many studies have indicated the OLA （ovine lymphocyte antigen） DRB1 gene is associated with some sheep diseases. Tibetan sheep is one of the three major shag sheep breeds in China, and also have the largest number of China＇s sheep breeds. But till now no report has been seen on studying DRB1 gene in Tibetan sheep of China. To understand the evolution and provide the basis for sheep disease resistance, polymorphism in the exon2 ofDRB1 gene in Tibetan sheep was analyzed. The PCR-SSCP, cloning and sequencing were used to analyse DRB1 gene variation in 600 Tibetan sheep of China. And the genetic relationship and evolutionary significance of the alleles had also been analyzed. Total of 31 alleles were identified, in which 15 alleles had not been reported before. And there were 70 SNPs （single nucleotide polymorphisms） sites in 31 sheep DRB1 gene haplotypes, the proportion was 29.5% to the whole exort2 sequence. All of this indicated that DRB1 exon2 is highly polymorphic in Tibetan sheep. The variation identified here might have an impact on both the function and level of expression of the OLA-DRB1.