目的 比较分析儿童腺病毒肺炎的常见实验室检查结果.方法 回顾性分析2017年12月-2019年3月广州市第一人民医院诊断为腺病毒肺炎的84例住院患儿,将其分为3型腺病毒感染组(n=36)和7型腺病毒感染组(n=37),轻症腺病毒肺炎组(n=56)和重症腺病毒肺炎组(n=28),分析比较两组间实验室检查结果.结果 幼龄及7型腺病毒与重症肺炎显著相关,7型腺病毒感染组的白细胞(WBC)、血小板(PLT)、血红蛋白(HGB)、肌酐(Cr)、尿素氮(BUN)、碱性磷酸酶(ALP)及钙(Ca)水平均低于3型腺病毒感染组,而血清天门冬氨酸氨基转移酶(AST)、肌酸激酶(CK)、乳酸脱氢酶(LDH)及α-羟丁酸脱氢酶(HBDH)水平均高于3型腺病毒感染组,差异有统计学意义(P＜0.05);重症腺病毒肺炎组Ca、HGB及ALP水平显著低于轻症腺病毒肺炎组,血清丙氨酸氨基转移酶(ALT)、γ-谷氨酰转移酶(GGT)、AST、LDH、HBDH、C反应蛋白(CRP)水平以及出现血尿、蛋白尿的频率均高于轻症腺病毒肺炎组,差异有统计学意义(P＜0.05).结论 重症腺病毒肺炎早期主要出现贫血及酶学异常升高,临床要重视这些指标的检查.

目的 比较分析腺病毒肺炎儿童中白细胞计数、C反应蛋白及降钙素原水平的差异变化,为腺病毒肺炎诊疗提供参考依据.方法 选取2017年12月至2019年3月诊断为腺病毒肺炎的患儿87例,将其分为3型肺炎组(38例)和7型肺炎组(37例),轻症肺炎组(57例)和重症肺炎组(30例),分析比较组间白细胞计数、C反应蛋白及降钙素原浓度.结果 7型肺炎组白细胞计数显著低于3型肺炎组[分别为(8.9±4.4)×109/L、(13.4±6.3)× 109/L,P＜0.01],而降钙素原浓度显著高于3型肺炎组[分别为3.37(0.38,4.03)、0.18(0.11,0.75) ng/mL,P＜ 0.01].重症肺炎组C反应蛋白和降钙素原浓度都高于轻症肺炎组[分别为50.3(14.1,63.2)、26.4(3.0,34.0)mg/L;分别为3.99(0.92,5.04)、0.28(0.11,0.8)ng/mL)],差异有统计学意义(P＜0.05).结论 C反应蛋白和降钙素原浓度的升高与重症腺病毒肺炎密切相关,可以作为评估腺病毒肺炎病情的可靠指标.

The whole-genome sequencing (WGS) of human adenoviruses (HAdVs) plays an important role in identifying, typing, and mutation analysis of HAdVs. Nowadays, three generations of sequencing have been developed. The accuracy of first-generation sequencing is up to 99.99%, whereas this technology relies on PCR and is time consuming; the next-generation sequencing (NGS) is expensive and not cost effective for determining a few special samples; and the third-generation sequencing technology has a higher error rate. In this study, first, we developed an efficient HAdV genomic DNA extraction method. Using the complete genomic DNA instead of the PCR amplicons as the direct sequencing template and a set of walking primers, we developed the HAdV WGS method based on first-generation sequencing. The HAdV whole genomes were effectively sequenced by a set of one-way sequencing primers designed, which reduced the sequencing time and cost. More importantly, high sequence accuracy is guaranteed. Four HAdV strains (GZ01, GZ02, HK35, and HK91) were isolated from children with acute respiratory diseases (ARDs), and the complete genomes were sequenced using this method. The accurate sequences of the whole inverted terminal repeats (ITRs) at both ends of the HAdV genomes were also acquired. The genome sequence of human adenovirus type 14 (HAdV-B14) strain GZ01 acquired by this method is identical to the sequence released in GenBank, which indicates that this novel sequencing method has high accuracy. The comparative genomic analysis identified that strain GZ02 isolated in September 2010 had the identical genomic sequence with the HAdV-B14 strain GZ01 (October 2010). Therefore, strain GZ02 is the first HAdV-B14 isolate emergent in China (September 2010; GenBank acc no. MW692349). The WGS of HAdV-C2 strain HK91 and HAdV-E4 strain HK35 isolated from children with acute respiratory disease in Hong Kong were also determined by this sequencing method. In conclusion, this WGS method is fast, accurate, and universal for common human adenovirus species B, C, and E. The sequencing strategy may also be applied to the WGS of the other DNA viruses.

The trivalent seasonal influenza vaccine was the only approved and available vaccine during the 2016-2018 influenza seasons. It did not include the B/Yamagata strain. In this study, we report an acute respiratory disease outbreak associated with influenza B/Yamagata infections in Guangzhou, Southern China (January through March, 2018). Among the 9914 patients, 2241 (22.6%) were positive for the influenza B virus, with only 312 (3.1%) positive for the influenza A virus. The influenza B/Yamagata lineage dominated during this period in Southern China. The highest incidence of influenza A virus infection occurred in the children aged 5-14 years. In contrast, populations across all age groups were susceptible to the influenza B virus. Phylogenetic, mutations, and 3D structure analyses of hemagglutinin (HA) genes were performed to assess the vaccine-virus relatedness. The recommended A/H1N1 vaccine strain (A/Michigan/45/2015) during both 2017-2018 and 2018-2019 was antigen-specific for these circulating isolates (clade 6B.1) in Spring 2018. An outbreak of influenza B/Yamagata (clade 3) infections in 2018 occurred during the absence of the corresponding vaccine during 2016-2018. The recommended influenza B/Yamagata vaccine strain (B/Phuket/3073/2013) for the following season (2018-2019) was antigen-specific. Although there were only a few influenza B/Victoria infections in Spring 2018, five amino acid mutations were identified in the HA antigenic sites of the 19 B/Victoria isolates (clade 1A), when compared with the 2016-2018 B/Victoria vaccine strain. The number was larger than expected and suggested that the influenza B HA gene may be more variable than previously thought. One of the mutations (K180N) was noted to likely alter the epitope and to potentially affect the viral antigenicity. Seven mutations were also identified in the HA antigenic sites of 2018-2020 B/Victoria vaccine strain, of which some or all may reduce immunogenicity and the protective efficacy of the vaccine, perhaps leading to more outbreaks in subsequent seasons. The combined epidemiological, phylogenetic, mutations, and 3D structural analyses of the HA genes of influenza strains reported here contribute to the understanding and evaluation of how HA mutations affect vaccine efficacy, as well as to providing important data for screening and selecting more specific, appropriate, and effective influenza vaccine candidate strains.

BACKGROUND Ovarian cancer is usually not diagnosed until the late stage, and it is resistant to platinum and other standard chemotherapy drugs, resulting in high mortality. OBJECTIVES To investigate the effects of miR-140-5p on cell proliferation, apoptosis, invasion, and migration capability in the SKOV3 and OVCAR3 ovarian cancer cell lines through Wnt signaling pathway. MATERIAL AND METHODS Expression levels of miR-140-5p were checked using quantitative real-time polymerase chain reaction (qRT-PCR). The expression of miR-140-5p was upregulated by transfecting cells with a miR-140-5p mimic or a mimic negative control (NC). Cell proliferation was assessed using a CCK8 assay, and cell cycle distribution and apoptosis percentage were detected with flow cytometry. A transwell invasion assay was employed to evaluate cell migration and invasion ability. The target complementary relationship between miR-140-5p and WNT1 was verified using a dual-luciferase reporter assay while â-catenin in the nuclei was observed using immunofluorescence. The expression of Wnt signaling pathway-related proteins was examined using western blot and qRT-PCR. RESULTS The relative expression level of miR-140-5p in SKOV3 and OVCAR3 cells was obviously decreased compared with that in the IOSE80 cells (p < 0.05). Besides, upregulated miR-140-5p effectively suppressed cell proliferation and increased the apoptosis ratio of SKOV3 and OVCAR3 cells (p < 0.05). In addition, the invasion and migration capability of SKOV3 and OVCAR3 cells in miR-140-5p mimic group was largely suppressed compared with the NC group (p < 0.05). What is more, the target complementary relationship between miR-140-5p and the WNT1 gene was revealed; upregulated miR-140-5p suppressed the expression of Wnt signaling-related genes, and restrained nuclear transfer of â-catenin (p < 0.05). CONCLUSIONS The overexpression of miR-140-5p restricted the proliferation, migration and invasion abilities, and accelerated cell apoptosis in ovarian cancer cell lines SKOV3 and OVCAR3 through the Wnt signaling pathway.

Objective M icroRNAs (MiRNAs) is thought to play a critical role in the initiation and progress of ovarian cancer (OC). Although miRNAs has been widely recognized in ovarian cancer, the role of hsa-miR-30a-5p (miR-30a) in OC has not been fully elucidated. Methods Three mRNA datasets of normal ovarian tissue and OC, GSE18520,GSE14407 and GSE36668, were downloaded from Gene Expression Omnibus (GEO) to find the differentially expressed gene (DEG). Then the target genes of hsa-miR-30a-5p were predicted by miRWALK3.0 and TargetScan. Then, the gene overlap between DEG and the predicted target genes of miR-30a in OC was analyzed by Gene Ontology (GO) enrichment analysis. Protein-protein interaction (PPI) network was conducted by STRING and Cytoscape, and the effect of HUB gene on the outcome of OC was analyzed. Results A common pattern of up-regulation of miR-30a in OC was found. A total of 225 DEG, were identified, both OC-related and miR-30a-related. Many DEG are enriched in the interactions of intracellular matrix tissue, ion binding and biological process regulation. Among the 10 major Hub genes analyzed by PPI, five Hub genes were significantly related to the overall poor survival of OC patients, in which the low expression of ESR1,MAPK10, Tp53 and the high expression of YKT,NSF were related to poor prognosis of OC. Conclusion Our results indicate that miR-30a is of significance for the biological progress of OC.

BACKGROUND:Acute coronary syndrome (ACS) is the leading cause of death in developing and developed countries, yet assessing the risk of its development remains challenging. Several lines of evidence indicate that small, dense low-density lipoproteins (sd-LDL) are associated with increased cardiovascular disease risk. We aim to evaluate sd-LDL concentration for predicting the risk of ACS in Chinese population. METHODS:Baseline characteristics of 121 patients with ACS and 172 healthy controls were obtained. Plasma sd-LDL-C was measured using homogeneous assay, and the proportion of sd-LDL-C in LDL-C was detected. RESULTS:There was gender and age effect on the sd-LDL-C concentration and sd-LDL-C/LDL-C ratio among healthy subjects. Elevated sd-LDL-C concentrations and sd-LDL-C/LDL-C ratio were observed in ACS patients with unstable angina pectoris (UAP), non-ST-segment elevation myocardial infarction (STEMI), and ST-segment elevation myocardial infarction (NSTEMI) compared with healthy controls (P < .05); however, there were no differences among ACS groups. According to Pearson's correlation coefficient analyses, sd-LDL-C concentration and sd-LDL-C/LDL-C ratio were positively correlated with triglyceride (TG) and LDL-C concentrations (P < .05) and negatively correlated with high-density lipoprotein (HDL) concentration (P < .05). Based on the receiver operating characteristic (ROC) curves, the cutoff values of sd-LDL-C and sd-LDL-C/LDL-C ratio for the prediction of ACS were 1.06 mmol/L and 34.55%, respectively. Multivariate logistic regression analysis demonstrated that the sd-LDL-C/LDL-C ratio, but not sd-LDL-C concentration, was significantly associated with ACS events [OR (95% CI): 1.24, 1.11-1.38, P < .001]. CONCLUSIONS:The sd-LDL-C/LDL-C ratio may be associated with an increased risk of developing ACS in Chinese population.

OBJECTIVE:To explore whether micro ribonucleic acid (miR)-7 affects the resistance of breast cancer cells to adriamycin (ADR) through regulating the epidermal growth factor receptor (EGFR)/phosphatidylinositol 3-hydroxy kinase (PI3K) signaling pathway.MATERIALS AND METHODS:MiR-7 expression was compared among MCF-10A, MCF-7 and MCF-7/ADR cells. The MCF-7/ADR cells were divided into three groups, namely miR-7 control group (MCF-7/ADR drug-resistant strains), miR-7 inhibition group (miR-7-inhibited MCF-7/ADR drug-resistant strains) and miR-7 promotion group (MCF-7/ADR drug-resistant strains transfected with miR-7), and the messenger RNA (mRNA) and protein expression levels of MCF-7/ADR were evaluated via Western blotting.RESULTS:The expression level of miR-7 was substantially decreased in MCF-7 and MCF-7/ADR cells (p<0.05), and it was lowered more obviously in MCF-7/ADR cells than that in MCF-7 cells (p<0.05). Compared with that in miR-7 control group, miR-7 expression in miR-7 promotion group was notably raised (p<0.05), proving that the sensitivity of MCF-7/ADR cells to ADR was enhanced, while that in miR-7 inhibition group was significantly lowered (p<0.05). Compared with those in miR-7 control group, the mRNA and protein expression levels of EGFR and PI3K were elevated in miR-7 inhibition group (p<0.05), while they were lowered in miR-7 promotion group (p<0.05). Additionally, compared with those in miR-7 control group, the proliferation and apoptosis abilities of cells in miR-7 inhibition group were markedly enhanced (p<0.05) and weakened (p<0.05), respectively, while they were weakened (p<0.05) and significantly strengthened (p<0.05), respectively in miR-7 inhibition group.CONCLUSIONS:MiR-7 plays an important role in the resistance of breast cancer cells to ADR, and its over-expression can inhibit the EGFR/PI3K signaling pathway to raise their sensitivity to the chemotherapy drug ADR.

OBJECTIVE:microRNAs are regulatory molecules regarded as important in the pathogenesis of different types of tumors. microRNA-216a (miR-216a-5p) has been identified as a tumor suppressor in multiple malignancies. However, the role of miR-216a-5p in the pathogenesis of small cell lung cancer (SCLC) remains obscure. The objective of this study was to investigate the role of the miR-216a-5p/Bcl-2 axis in SCLC pathogenesis.MATERIALS AND METHODS:All the experimental methods used were as follows: microarray analysis, cell culture, transient, and stable gene transfection; real-time fluorescence PCR; Western blot; flow cytometry for cell cycle analysis; in vitro proliferation assay; in vitro wound healing experiment; in vivo xenograft model in nude mice; and dual luciferase reporter assay. All statistical analyses were carried out using GraphPad Prism 7 software. Statistical significance was analyzed by Student's t-test or one-way ANOVA. P <0.05 (typically compared with the negative control group) was considered as significant and is marked with an asterisk in the figures.RESULTS:In this study, we observed that miR-216a-5p is downregulated in SCLC cell lines compared to that in the normal human bronchial epithelial cell line 16-HBE. In vitro and in vivo experiments demonstrate that upregulation of miR-216a-5p significantly decreased cell growth and migration and its downregulation increased SCLC cell proliferation and migration and influenced the cell cycle. Using bioinformatics analyses, we predicted that the important antiapoptotic gene Bcl-2 is targeted by miR-216a-5p, and we identified a functional miR-216a-5p binding site in the 3'-UTR of Bcl-2 using luciferase reporter assay. Furthermore, we determined that suppression of miR-216a-5p modulated the expression of Bcl-2, Bax, and Bad proteins (Bcl-2 family proteins), while Bcl-2 knockdown abrogated the effect of miR-216a-5p downregulation on cell proliferation, cell migration, and the cell cycle.CONCLUSION:Taken together, these findings suggest that miR-216a-5p regulates SCLC biology via Bcl-2 family proteins. Therefore, our study highlights the role of the miR-216a-5p/Bcl-2 axis in SCLC pathogenesis.

Human adenovirus type 14 (HAdV-B14p) was originally identified as an acute respiratory disease (ARD) pathogen in The Netherlands in 1955. For approximately fifty years, few sporadic infections were observed. In 2005, HAdV-B14p1, a genomic variant, re-emerged and was associated with several large ARD outbreaks across the U.S. and, subsequently, in Canada, the U.K., Ireland, and China. This strain was associated with an unusually higher fatality rate than previously reported for both this prototype and other HAdV types in general. In China, HAdV-B14 was first observed in 2010, when two unrelated HAdV-B14-associated ARD cases were reported in Southern China (GZ01) and Northern China (BJ430), followed by three subsequent outbreaks. While comparative genomic analysis, including indel analysis, shows that the three China isolates, with whole genome data available, are similar to the de Wit prototype, all are divergent from the U.S. strain (303600; 2007). Although the genomes of strains GZ01 and BJ430 are nearly identical, as per their genome type characterization and percent identities, they are subtly divergent in their genome mutation patterns. These genomes indicate possibly two lineages of HAdV-B14 and independent introductions into China from abroad, or subsequent divergence from one; CHN2012 likely represents a separate sub-lineage. Observations of these simultaneously reported emergent strains in China add to the understanding of the circulation, epidemiology, and evolution of these HAdV pathogens, as well as provide a foundation for developing effective vaccines and public health strategies, including nationwide surveillance in anticipation of larger outbreaks with potentially higher fatality rates associated with HAdV-B14p1.

Objective To study the viral etiologic characteristics of acute respiratory viruses associated with adults influenza like illness (ILI) case.Methods ILI cases and non-ILI cases of nose swabs specimen were collected from Zhujiang Hospital of Southern Medical University from Nov 2009 to Oct 2014.qRT-PCR was used to test for 18 respiratory viruses.Results 201 virus positive responses were detected in samples from 420 (47.9％) patients with influenza like illness,while only 3 cases (2.6％) were found to be infected with virus in 115 non-ILI cases.The most common virus identified was influenza virus (IFV) in frequency of 51.2％ (103/201),followed by human rhinovirus (RHV) and coronaviruses (Coy).There was no significant difference among each age group in virus detection rate,however,there were obvious differences in viruses detected of several age groups,which showed that the main inflect crowd of influenza was 18 to 30 age group,the main inflect crowd of human coronavirus was 51-77 age group,and the main inflect crowd of human rhinovirus was 51-90 age group.Conclusion ILI cases in adult mosdy were caused by infection with several kinds of respiratory virus,and the dominant virus for ILI was proved to be the seasonal influenza viruses.

Background Our study investigated 2 common single-nucleotide polymorphisms (SNPs) of vascular endothelial growth factor (VEGF) for their influences on serum VEGF levels, disease activity, and synovial lesions in rheumatoid arthritis (RA). Material/Methods Clinical information and venous blood samples were collected from 98 RA patients and 100 healthy controls. Genotyping on samples from the subjects was performed using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). Serum VEGF levels were determined using the enzyme-linked immunosorbent assay (ELISA). The synovial thickness and joint effusion of 28 joints were measured in RA patients, and total sharp score (TSS) and disease activity score (DAS) of 28 joints were recorded. Results The genotype and allele frequencies of VEGF rs833070 (G>A) and rs3025030 (G>C) were significantly different between RA group and control group (all P<0.05). VEGF rs833070 and rs3025030 polymorphisms were associated with increasing VEGF serum levels in the RA group (all P<0.01). Statistically significant difference was observed in DAS28 between the different genotypes of VEGF rs833070 in RA patients (P<0.05). Importantly, significant differences in synovial thickening, joint effusion and synovial angiogenesis were observed between the different genotypes of VEGF rs833070 and rs3025030 polymorphisms (all P<0.05). Conclusions Our study provides evidence that VEGF polymorphisms might be important indicators of disease activity and synovial lesions, and prognostic factors in evaluating the treatment effectiveness in RA.

Human adenoviruses (HAdVs) are highly contagious pathogens causing acute respiratory disease (ARD), such as community-acquired pneumonia. HAdV-7d, a re-emergent genomic variant, has been recently reported in Asia and the United States after a several-decade absence. However, whether HAdV-7d is associated with higher severity than other types is currently unclear. In this study, the clinical and epidemiological investigation showed that fever, cough, and sore throat were the three most common respiratory symptoms of HAdV infections. HAdV-7 caused longer duration of fever, higher morbidity of tachypnea/dyspnea, pleural effusion, diarrhea, hepatosplenomegaly, consciousness alteration, as well as higher rates of pneumonia, mechanical ventilation and higher fatality rate (28.6%) than other types, particularly HAdV-3 and HAdV-2. The genomes of seven HAdV-7d isolates from mild, severe, and fatal cases were sequenced and highly similar with each other. Surprisingly, two isolates (2011, 2012) had 100% identical genomes with an earlier strain from a fatal ARD outbreak in China (2009), which elucidates the virus origin and confirms the unexpected HAdV genomic conservation and stability. Phylogenetic analysis indicated that L1 52/55-kDa DNA packaging protein may be associated with the higher severity of illness and fatality rate of HAdV-7. Clinicians need to be aware of HAdVs in children with ARD.

目的 建立两种不同循环参数的实时荧光定量PCR,检测包括流感病毒、冠状病毒等18种(型/亚型)呼吸道病毒.方法 引用文献报道的18种(型/亚型)呼吸道病毒引物和探针,建立两种不同循环参数的实时荧光定量PCR法,考核方法的敏感性、特异性和准确性,并用临床标本进行验证.结果 成功建立了可检测18种(型/亚型)呼吸道病毒的两种不同循环参数的实时荧光定量PCR法;该法特异性较好,最低检测限为10拷贝数/μL,变异系数(CV)为1.26％ ～ 3.43％,重复性良好;408份临床标本中,用该法检测鼻拭子标本阳性率为63.2％(225/356),肺泡灌洗液标本阳性率为25.0％(13/52).结论 建立的实时荧光定量PCR法简便、特异、敏感、稳定,适用于临床常见呼吸道病毒感染的早期诊断.