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In addition to the typical respiratory response, new coronavirus disease 2019 (COVID-19) is also associated with very common gastrointestinal symptoms. Cases with gastrointestinal symptoms are more likely to be complicated by liver injury and acute respiratory distress syndrome (ARDS). If not treated in time, coma and circulatory failure may ensue. As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects the human body through the combination of angiotensin-converting enzyme 2 (ACE2) in the gastrointestinal tract, the mechanism underlying the gastrointestinal symptoms may involve damage to the intestinal mucosal barrier and promotion of the production of inflammatory factors. Indeed, after cells in the lungs become infected by SARS-CoV-2, effector CD4+ T cells reach the small intestine through the gut-lung axis, causing intestinal immune damage and diarrhea; early extensive use of antibacterial and antiviral drugs can also lead to diarrhea in patients. Thus, treatment options for COVID-19 patients should be promptly adjusted when they have gastrointestinal symptoms. As SARS-CoV-2 has been detected in the feces of COVID-19 patients, future prevention and control efforts must consider the possibility of fecal-oral transmission of the virus.

This study investigated whether the rotavirus infection rate in children is associated with temperature and air pollutants in Hangzhou, China. This study applied a distributed lag non-linear model (DLNM) to assess the effects of daily meteorological data and air pollutants on the rotavirus positive rate among outpatient children. There was a negative correlation between temperature and the rotavirus infection rate. The impact of temperature on the detection rate of rotavirus presented an evident lag effect, the temperature change shows the greatest impact on the detection rate of rotavirus approximate at lag one day, and the maximum relative risk (RR) was approximately 1.3. In 2015, the maximum cumulative RR due to the cumulative effect caused by the temperature drop was 2.5. Particulate matter (PM) 2.5 and PM10 were the primary air pollutants in Hangzhou. The highest RR of rotavirus infection occurred at lag 1–1.5 days after the increase in the concentration of these pollutants, and the RR increased gradually with the increase in concentration. Based on the average concentrations of PM2.5 of 53.9 μg/m 3 and PM10 of 80.6 μg/m 3 in Hangzhou in 2015, the cumulative RR caused by the cumulative effect was 2.5 and 2.2, respectively. The current study suggests that temperature is an important factor impacting the rotavirus infection rate of children in Hangzhou. Air pollutants significantly increased the risk of rotavirus infection, and dosage, lag and cumulative effects were observed.

This study investigated whether respiratory syncytial virus (RSV) infection in children was associated with ambient temperature and air pollutants in Hangzhou, China. A distributed lag non-linear model (DLNM) was used to estimate the effects of daily meteorological data and air pollutants on the incidence of RSV infection among children. A total of 3650 childhood RSV infection cases were included in the study. The highest air pollutant concentrations were in January to May and October to December during the year. The yearly RSV-positive rate was 10.0 % among children with an average age of 4.3 months. The highest RSV-positive rate occurred among patients 0 to 3 months old. Children under 6.5 months old accounted for 80 % of the total patients infected by RSV. A negative correlation was found between ambient temperature and RSV infection, and it was strongest with minimum ambient temperature (r = −0.804, P

Resumen Introduccion El asma es una afeccion inflamatoria de las vias respiratorias. Las infecciones por Mycoplasma pneumoniae pueden exacerbar los sintomas del asma. Se ha demostrado que la interleucina 2 y la interleucina 4 participan en las reacciones inmunitarias e inflamatorias. Hemos estudiado la relacion entre los polimorfismos de la IL2 y la IL4 y su expresion y el riesgo de padecer asma e infeccion por M. pneumoniae en ninos. Metodos Se recluto a 392 ninos asmaticos y 849 controles para el estudio. Se genotiparon 8 polimorfismos en IL2 e IL4 con la plataforma MassARRAY de Sequenom. La infeccion por M. pneumoniae y el numero de copias se establecieron mediante PCR fluorescente. Los niveles sericos de expresion de IL-2 e IL-4 se midieron con ELISA. Resultados Hallamos una relacion significativa entre el polimorfismo rs6534349 de IL2 y el aumento de riesgo de sufrir asma (heterocigoticos, p = 0,029; variantes homocigoticas, p = 0,013), asi como entre el polimorfismo rs2227284 de IL4 y una reduccion del riesgo de padecer asma (heterocigoticos, p = 0,026; variantes homocigoticas, p = 0,001). Ademas, la relacion con otros polimorfismos, excepto el rs2070874, se hizo evidente al agrupar a los ninos asmaticos segun la clasificacion GINA de control y gravedad del asma. Asimismo, los niveles sericos de expresion de IL-2 e IL-4 fueron significativamente mayores en los sujetos no infectados (p = 0,038) e infectados (p = 0,011) por M. pneumoniae, respectivamente. Esta observacion tambien se cumple entre los pacientes asmaticos (p = 0,016 para IL-2 y p = 0,042 para IL-4), pero en los controles no asmaticos solo se cumple en el caso de la IL-4 (p = 0,032). Del mismo modo, observamos que el genotipo GG rs6534349 estaba claramente relacionado con un aumento de las posibilidades de tener una infeccion con alta carga de M. pneumoniae (p = 0,0376). Conclusiones La IL2 y la IL4 podrian ser biomarcadores importantes para calcular el riesgo de padecer asma, asi como infeccion por M. pneumoniae, en ninos.

Objective Kawasaki disease (KD) is the primary cause of heart disease among children, but because its clinical symptoms are nonspecific, it is difficult to diagnose. The purpose of this study was to evaluate laboratory indices for possible use in the early diagnosis of KD and to determine which indices are predictive of a response to intravenous immunoglobulin (IVIG) and can be used to monitor the effects of treatment. Methods Three hundred thirty KD patients, 330 age-matched children with KD-like febrile disease, and 330 age-matched healthy children (controls) were enrolled in this prospective study. Levels of N-terminal pro–brain natriuretic peptide (NT-proBNP), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and cytokines were determined in all study subjects. Results In the derivation cohort, 181 patients in the KD group were compared with 181 patients in the KD-like febrile group. The following indices were found to be useful in the diagnosis of KD: NT-proBNP (area under the curve [AUC] 0.923), ESR (AUC 0.909), CRP (AUC 0.834), and interleukin-6 (IL-6; AUC 0.678). The diagnostic efficiency of each index demonstrated in the derivation cohort was repeated in the 149 KD patients in the validation cohort. There were significant differences in NT-proBNP levels between IVIG-responsive KD patients (n = 270) and IVIG-nonresponsive KD patients (n = 60), with higher NT-proBNP levels in IVIG-nonresponsive KD patients. The NT-proBNP level can effectively distinguish IVIG-responsive KD patients from IVIG-nonresponsive patients, and its AUC was 0.73. There were also significant differences in the NT-proBNP levels before and after treatment, with a significant decline after treatment. Conclusion Serum levels of NT-proBNP can be used in the diagnosis of KD, the prediction of a patient's sensitivity to IVIG treatment, and the monitoring of the effects of IVIG treatment, but more attention must be paid to the scope of its application.

Autophagy is a dynamic process accomplished by the generation and maturation of autophagosomes. Isolation of autophagosomes and subsequent compositional analysis can provide information about their biogenesis mechanism. In this article, HEK293 cells expressing GFP-LC3 were treated by calcium phosphate precipitates (CPP) to induce autophagy. The autophaogomes induced by CPP were tubular and vesicular structures, extensively formed in the cytosol. After all membranes in the cell lysate were fractionated by differential centrifugation, autophagosomes from light and heavy membranes were isolated by immuno-precipitation, using antibodies against GFP-LC3 and Atg5. We found that GFP-LC3 and Atg5 positive autophagosomes represented distinctive subpopulations. Judged from the molecular markers associated, including organelle markers and Atg proteins, GFP-LC3 positive autophagosomes were overall at the later biogenetic stage. Furthermore, both GFP-LC3 and Atg5 positive autophagosomes from light membranes were less mature than those from heavy membranes. We have established a method to isolate subpopulations of autophagsomes for further characterization.

There is growing public concern regarding assay sensitivity to HBsAg mutants in clinical diagnosis and vaccine escape. The aim of this study is to introduce a new HBsAg mutant strain. The serum samples were those of patient X at the age of 3 months and 3 years respectively, and of her mother immediately before parturition, which were used to amplify the HBsAg-coding DNA fragments by PCR. The HBsAg DNA sequences were translated into their corresponding amino acid sequences and then aligned in pubmed with nucleotide blast. The sequencing data of S coding regions shows that patient X has been infected by a new HBV variant with an A to C substitution at nt431, resulting in an Asp(GAC)to Ala(GCC) substitution at aa144 of major protein; CC to AA substitution at nt359 and nt360, resulting in an Pro(CCC) to Gln(CAA) substitution at aa120 of pre “a” epitope; A to G substitution at nt491, resulting in an Glu(GAG) to Gly(GGG) substitution at aa164 of post “a” epitope. Three new mutations (S171F, S174N and Q181R) at the antigenic epitopes of HBV presented by HLA class I molecules are found. The HBV mutant strain causes vaccine escape and occult infection.

In order to develop a gene therapy to human cytomegalovirus (HCMV), RNA interference (RNAi) was employed to inhibit the expression of HCMV UL122 gene in vitro. Recombinant vector pUL122-EGFP, which expressed UL122-EGFP fusion protein, and recombinant vectors psi122-1, psi122-2 and psi122-3, which expressed small interfering RNAs (siRNAs) targeted to UL122 were contransfected into AD293 cells. The fluorescence signal of pUL122-EGFP was greatly suppressed by psi122-1 and psi122-2, with an inhibitory rate of 82.0% ± 1.0% and 79.5% ± 2.5%, respectively. The mRNA of pUL122-EGFP of the cells transfected with psi122-1 and psi122-2 was decreased 97.3% ± 0.6% and 98.0% ± 0.1%, respectively. Vector psi122-3 showed a slightly low suppression rate. Therefore, it may be concluded that plasmids encoding siRNAs targeted to UL122 is able to in vitro reduce markedly the expression of UL122-EGFP. And it is very likely that the psi122-1 and psi122-2 are potentially efficacious siRNAs in the gene therapy of HCMV infection in vivo, in which further investigations are required. This study is expected to greatly facilitate the use of the RNAi technology for the anti-HCMV studies. (© 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

A method for the detection of bacterial pathogens in sepsis and bacterial meningitis with 16S rRNA gene— based real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) is developed. A total of 190 blood specimens and 5 cerebrospinal fluid specimens from neonates with suspected sepsis or bacterial meningitis were evaluated with 16S rRNA gene—based real-time FQ-PCR assay. The positive rate of the real-time FQ-PCR assay was significantly higher (25/195, 12.82%) than that of bacterial culture (15/195, 7.69%; P = .002). When bacterial culture was used as a control, the sensitivity of the real-time FQ-PCR was 100%, the specificity was 94.4%, and Youden's index was 0.944. This study suggests that 16S rRNA gene—based real-time FQ-PCR assay is an important and accurate method in the detection of bacterial pathogens of sepsis and bacterial meningitis and should have a promising usage in the diagnosis of sepsis and bacterial meningitis.

The aim of the study was to develop a multiplex PCR-based DNA microarray technology for simultaneous detection and species identification of seven human herpes viruses, namely herpes simplex virus type 1, type 2 (HSV-1, HSV-2), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpes virus 6 (HHV-6A, HHV-6B), and to apply this technology to accurate diagnosis of herpesvirus-associated diseases. Primers and oligonucleotide probes were designed and synthesized based on the highly conserved regions of the DNA polymerase gene in human herpes viruses. DNA microarrays were made by printing the oligonucleotide probes onto special glass slides. After amplification and labeling with CY5, the PCR products were hybridized with the DNA microarrays and species identified. Sixty-one cerebrospinal fluid (CSF) and 132 blood specimens were analyzed by this technique, and the results were compared with those of TaqMan PCR. Several specimens were sequenced further after cloning. The PCR products of the seven human herpes viruses ranged from 224 to 252 bp, and could be species identified with DNA microarrays. The detection limits were 10(1) copies/microl for each virus. And the test showed no cross-reaction to DNA extracted from S. aureus, E. coli, hepatitis B virus, Cryptococcus neoformans, Candida albicans and human genome. Among 132 blood and 61 CSF specimens, 55 were tested positive for human herpes virus DNA. Compared with the results of TaqMan PCR, the sensitivity and specificity of the DNA microarray technology was 96.2% and 99.3%, respectively. This multiplex PCR-based DNA microarray technology, which is rapid, specific and sensitive, serves as an effective technique for simultaneous detection and species identification of seven human herpes viruses.

In order to develop a gene therapy to human cytomegalovirus (HCMV), RNA interference (RNAi) was employed to inhibit the expression of HCMV UL122 gene in vitro. Recombinant vector pUL122-EGFP, which expressed UL122-EGFP fusion protein, and recombinant vectors psi122-1, psi122-2 and psi122-3, which expressed small interfering RNAs (siRNAs) targeted to UL122 were contransfected into AD293 cells. The fluorescence signal of pUL122-EGFP was greatly suppressed by psi122-1 and psi122-2, with an inhibitory rate of 82.0% +/- 1.0% and 79.5% +/- 2.5%, respectively. The mRNA of pUL122-EGFP of the cells transfected with psi122-1 and psi122-2 was decreased 97.3% +/- 0.6% and 98.0% +/- 0.1%, respectively. Vector psi122-3 showed a slightly low suppression rate. Therefore, it may be concluded that plasmids encoding siRNAs targeted to UL122 is able to in vitro reduce markedly the expression of UL122-EGFP. And it is very likely that the psi122-1 and psi122-2 are potentially efficacious siRNAs in the gene therapy of HCMV infection in vivo, in which further investigations are required. This study is expected to greatly facilitate the use of the RNAi technology for the anti-HCMV studies.