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[This corrects the article DOI: 10.1016/j.apsb.2021.04.013.].
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Congenital hydrocephalus is a major neurological disorder with high rates of morbidity and mortality; however, the underlying cellular and molecular mechanisms remain largely unknown. Reproducible animal models mirroring both embryonic and postnatal hydrocephalus are also limited. Here, we describe a new mouse model of congenital hydrocephalus through knockout of β-catenin in Nkx2.1-expressing regional neural progenitors. Progressive ventriculomegaly and an enlarged brain were consistently observed in knockout mice from embryonic day 12.5 through to adulthood. Transcriptome profiling revealed severe dysfunctions in progenitor maintenance in the ventricular zone and therefore in cilium biogenesis after β-catenin knockout. Histological analyses also revealed an aberrant neuronal layout in both the ventral and dorsal telencephalon in hydrocephalic mice at both embryonic and postnatal stages. Thus, knockout of β-catenin in regional neural progenitors leads to congenital hydrocephalus and provides a reproducible animal model for studying pathological changes and developing therapeutic interventions for this devastating disease.
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Animals , Mice , Disease Models, Animal , Hydrocephalus/genetics , Mice, Knockout , Neurons , beta Catenin/geneticsABSTRACT
Objective:To provide data support for the prevention and control of dengue fever in Henan Province by analyzing the molecular epidemiological and etiological characteristics of dengue fever outbreaks in Puyang in 2019.Methods:Blood samples were collected from all suspected cases of dengue fever. The antigen, antibody and nucleic acid of dengue virus (DENV) were detected. The E gene was amplified by viral nucleic acid extraction and sequenced. Phylogenetic tree was constructed to trace the source of infection. Results:A total of 61 local cases of dengue fever were reported, and no deaths were reported. Among them, 4 cases (72.13%) were positive for DENV NS1 antigen; 16(26.23%) cases were positive for specific IgM; 38(62.30%) cases were positive for specific IgG; 34 cases (54.10%) were positive for dengue nucleic acid testing. Ten dengue virus strains were isolated, all of which were dengue virus type 1(DENV-1). Sequence analysis of E gene suggested it belonged to the same clade as Henan201903 strain imported from Cambodia to Zhumadian, Henan in 2019, with the highest homology. Conclusions:The dengue fever epidemic in Henan Province was caused by DENV-1, which might be improted from Cambodia, Singapore, Myanmar and other Southeast Asian countries. Therefore, the surveillance of DENV in people returning from Southeast Asia should be strengthened.
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Vascular smooth muscle cell (VSMC) migration plays a critical role in the pathogenesis of many cardiovascular diseases. We recently showed that TMEM16A is involved in hypertension-induced cerebrovascular remodeling. However, it is unclear whether this effect is related to the regulation of VSMC migration. Here, we investigated whether and how TMEM16A contributes to migration in basilar artery smooth muscle cells (BASMCs). We observed that AngII increased the migration of cultured BASMCs, which was markedly inhibited by overexpression of TMEM16A. TMEM16A overexpression inhibited AngII-induced RhoA/ROCK2 activation, and myosin light chain phosphatase (MLCP) and myosin light chain (MLC20) phosphorylation. But AngII-induced myosin light chain kinase (MLCK) activation was not affected by TMEM16A. Furthermore, a suppressed activation of integrin
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It is valuable to establish an efficient and accurate virus detection strategy. Although the examination of viral nucleic acid maybe the gold standard for diagnosing COVID-19, there are still many problems in related research. Therefore, utilizing the latest research technologies in the fields of molecular biology, cell biology, immunology to detect the virus in different types of clinical specimens, supplemented by information such as medical imaging, can help achieve early detection, isolation and treatment of patients.
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Objective:To analyze the neutralization properties of different genotypes and mutants of severe fever with thrombocytopenia syndrome virus (SFTSV).Methods:Pseudoviruses of SFTSV of different genotypes and mutants were constructed using VSVΔG-Fluc*G backbone. Neutralization assays were established based on the pseudoviruses. DNA vaccines for different SFTSV genotypes were prepared. Serum samples were collected from guinea pigs immunized with the DNA vaccines. Neutralizing antibodies in serum samples from immunized guinea pigs and naturally infected patients were detected using neutralization assays and analyzed.Results:The pseudoviruses of five genotypes and 43 mutants were successfully constructed and the neutralization assays based the pseudoviruses were successfully established after optimizing the reaction parameters. The dilution multiple corresponding to the inhibition rate of neutralizing antibody to half of the pseudovirus infection was taken as the titer of neutralizing antibody by the reduction in pseudovirus reporter gene. The neutralization antibody titers in naturally infected patients and immunized guinea pigs were respectively in the ranges of 1∶100-1∶43 000 and 1∶100-1∶2 500 when detected with the reference HB29 pseudovirus. The neutralization antibody titers ranged from 1∶100-1∶2 500 after immunization with different genotypes of DNA vaccines. No significant statistical difference in neutralization antibody titer was observed among different genotypes or mutant strains.Conclusions:The neutralization properties of different genotypes and mutants showed no significant change, which would be very useful for developing vaccines.
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Drosophila neural development undergoes extensive chromatin remodeling and precise epigenetic regulation. However, the roles of chromatin remodeling in establishment and maintenance of cell identity during cell fate transition remain enigmatic. Here, we compared the changes in gene expression, as well as the dynamics of nucleosome positioning and key histone modifications between the four major neural cell types during Drosophila neural development. We find that the neural progenitors can be separated from the terminally differentiated cells based on their gene expression profiles, whereas nucleosome distribution in the flanking regions of transcription start sites fails to identify the relationships between the progenitors and the differentiated cells. H3K27me3 signal in promoters and enhancers can not only distinguish the progenitors from the differentiated cells but also identify the differentiation path of the neural stem cells (NSCs) to the intermediate progenitor cells to the glial cells. In contrast, H3K9ac signal fails to identify the differentiation path, although it activates distinct sets of genes with neuron-specific and glia-related functions during the differentiation of the NSCs into neurons and glia, respectively. Together, our study provides novel insights into the crucial roles of chromatin remodeling in determining cell type during Drosophila neural development.
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Objective@#To confirm the laboratory diagnosis of dengue bordline cases reported in Henan Province and trace its origin from molecular level in 2017.@*Methods@#The study samples were blood samples (3-5 ml), which came from 8 suspected cases of dengue fever reported in the 2017 direct reporting system of Henan provincial infectious disease monitoring network. Meanwhile, case investigation was conducted according to National dengue fever surveillance programme. Serum were separated from blood samples and tested for Dengue NS1 antigen, IgM & IgG antibodies, and dengue RNA. According to dengue diagnosis criteria, confirmed cases were identified by testing results. Samples carried dengue RNA performed for real-time PCR genotyping and amplification of E gene. Then, the amplicons were sequenced and homological and phylogenetic analyses were constructed.@*Results@#8 serum samples of suspected dengue cases were collected in Henan Province, 2017. Six of them were diagnosed as dengue confirmed cases. All the dengue confirmed cases belonged to outside imported cases, 5 of them were positive by dengue RNA testing. Genotyping results showed there were 1 DENV1 case, 2 DENV2 cases and 2 DENV3 cases. A DENV2 case and a DENV3 case of this study were traced its origin successfully. The sequence of Pakistan imported DENV2 case belongs to cosmopolitan genotype, which was the most consistent with Pakistan's DENV2 KJ010186 in 2013 (identity 99.0%). The sequence of Malaysia imported DENV3 case belongs to genotype I, which was the most consistent with Singapore's DENV3 KX224276 in 2014(identity 99.0%).@*Conclusion@#The laboratory diagnosis and molecular traceability of dengue cases in Henan Province in 2017 confirmed that all cases were imported and did not cause local epidemics.
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Objective To analyze the VP1 sequences of coxsackievirus A16(CA16) causing neu-rologic complications. Methods Clinical samples and epidemiological information were collected from pa-tients with viral encephalitis. Coxsackievirus A16 in these samples were first detected with real time RT-PCR and then isolated. RT-PCR was performed to amplify VP1 sequences and the amplified products were se-quenced. DNAStar 5.0 and Mega 5 were used for sequence analysis. All data was analyzed with SPSS statis-tical software. Results Fifteen samples were collected from 12 patients with hand, foot and mouth disease (HFMD) complicated by neurologic complications. Eight patients had the symptoms of fever, skin rash, signs of meningeal irritation and neck rigidity. No typical cluster was associated with clinical features or the time of onset. Both pharyngeal/anal swab and serum samples were collected from three patients (patient′s number:01111,01169 and 01130). The two samples collected from both 01111 and 01130 patients shared 100% similarity in nucleotide and amino acid based on VP1 sequences,while those from 01169 patient dif-fered in only one base. The 15 CA16 isolates were highly similar in VP1 gene, sharing 94.5%-100% ho-mology in nucleotide sequences and 98.0%-100% homology in amino acid sequences. These 15 isolates showed 68.5%-70.5% identities in nucleotide sequences and 90.5%-91.9% identities in amino acid se-quences with the CA16 prototype strain G10. Phylogenetic analysis revealed that based upon VP1 sequences, all of the 15 CA16 isolates grouped into genotype B subtype 1b (B1b), which was further classified into three clusters. Conclusion All of the 15 CA16 isolates causing neurologic complications belonged to B1b sub-genotype. Understanding the molecular epidemiology of CA16 would be essential for controlling morbidi-ty rates of HFMD and vaccine research.
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Objective To analyze the genotyping of hantavirus and investigate the pathogenic features of local rats in Henan province. Methods A total of 600 rats captured in Queshan county, Zhumadian city from 2014-2016 were chosen to find out the major species and density. Rat lung specimens were detected by RT-PCR using partial M and S segment primers, then sequencing and phylogenetic analysis based on M segment (2003-2302 nt) were performed to analyze gene subtype and evolution. Results In the field of Queshan county, major species were sewer rats and apodemus agrarius, and the average density of rats was 1.33%-1.83%. Sewer rats, mus musculus and apodemus agrarius were major species in the residential area, and the average density of rats was 1.36%-1.97%. Hantaviruses were detected by RT-PCR in three captured rats in 2014, and the species were mus musculus, cricetulus triton and sewer rats. Nucleotide homology similarity based M and S segment of three positive products was 100%. Phylogenetic analysis indicated the virus was belonged to S4 subgenotype of Seoul virus, which was similar with the strains in Korea and Hubei province, China. Conclusion The virus from rats in Queshan county, Henan province is seoul virus, S4 subgenotype. It is necessary to take the relevant prevention and control measures to prevent hemorrhagic fever of renal syndrome because of wide host range.
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Objective To evaluate the psychometric characteristics of the Chinese version of Cataldo Lung Can cer Stigma Scale (CLCSS). Methods Convenience sampling was used to recruit 775 lung cancer patients in a tertiary hospital and some anticancer groups in Beijing,and the Chinese version of CLCSS was used to perform measure-ment. Results The S-CVI/UA was 0.87,and S-CVI/Ave was 0.96. Construct validity consisted of EFA and CFA. Through EFA,the Chinese version of CLCSS included four dimensions which accounted for 55.248% of the accu-mulated variance. Through CFA,all the indicators were in adaptation standard range. The test-retest reliability for the Chinese version of CLCSS was 0.801(P<0.01). Cronbach's α coefficient was 0.932,and Cronbach's α coeffi-cients for four dimensions ranged from 0.799 to 0.922. Conclusion The reliability and validity of the Chinese version of CLCSS are satisfactory and the Chinese version of CLCSS can be used among Chinese lung cancer patients.
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Objective To explore the characteristics regarding temporal,spatial and spatiotemporal distribution on severe fever with thrombocytopenia syndrome (SFTS) in Henan province.Methods Surveillance data related to SFTS was collected in Henan province,from year 2014 to 2016.Descriptive method was used to analyze the distribution of SFTS.1.7.0 software related to the Public health geographic information system (PHGIS),was applied to draw the spatial distribution map of SFTS.Chi-square test was used to compare the different incidence rates.Results A total of 2 781 SFTS cases,including 34 deaths,were reported in Henan province from 2014 to 2016,with an average annual fatality rate as 1.22％.There were statistically significant differences for the incidence rates of SFTS between different years (P＜0.01).Cases were mainly concentrated from April to October,which accounted for 96.66％ of the total number,with the incidence peak seen in May.Incidence rates of SFTS in spring,summer,autumn were higher than that in winter.The cases were scattering around in 26 counties of 8 cities.Xinyang city reported 2 714 cases,accounting for 97.59％ of the total number of cases in the province.The average annual incidence rate in Xinyang city was 17.22 per 100 000,much higher than that for the whole Henan province (0.98 per 100 000),with statistically significant difference (P＜0.01).Six counties reported having death cases,that accounted for 23.08％ of the total number of counties,reported to have death cases.Two kinds of incidence patterns of SFTS were noticed in Henan province,with aggregation in some local regions or sporadic in individual counties.The number of counties with reporting cases increased annually.The epidemic area was expanding and gradually spreading from south to north areas of the province.Conclusions SFTS was characterized with both temporal and spatial clusters in Henan province.Effective prevention and control measures should be made in accordance with the spatiotemporal distribution and the trend on SFTS.
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Objective To explore the characteristics regarding temporal,spatial and spatiotemporal distribution on severe fever with thrombocytopenia syndrome (SFTS) in Henan province.Methods Surveillance data related to SFTS was collected in Henan province,from year 2014 to 2016.Descriptive method was used to analyze the distribution of SFTS.1.7.0 software related to the Public health geographic information system (PHGIS),was applied to draw the spatial distribution map of SFTS.Chi-square test was used to compare the different incidence rates.Results A total of 2 781 SFTS cases,including 34 deaths,were reported in Henan province from 2014 to 2016,with an average annual fatality rate as 1.22％.There were statistically significant differences for the incidence rates of SFTS between different years (P＜0.01).Cases were mainly concentrated from April to October,which accounted for 96.66％ of the total number,with the incidence peak seen in May.Incidence rates of SFTS in spring,summer,autumn were higher than that in winter.The cases were scattering around in 26 counties of 8 cities.Xinyang city reported 2 714 cases,accounting for 97.59％ of the total number of cases in the province.The average annual incidence rate in Xinyang city was 17.22 per 100 000,much higher than that for the whole Henan province (0.98 per 100 000),with statistically significant difference (P＜0.01).Six counties reported having death cases,that accounted for 23.08％ of the total number of counties,reported to have death cases.Two kinds of incidence patterns of SFTS were noticed in Henan province,with aggregation in some local regions or sporadic in individual counties.The number of counties with reporting cases increased annually.The epidemic area was expanding and gradually spreading from south to north areas of the province.Conclusions SFTS was characterized with both temporal and spatial clusters in Henan province.Effective prevention and control measures should be made in accordance with the spatiotemporal distribution and the trend on SFTS.
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Objective To evaluate different detection methods in the diagnosis of severe fever with thrombocytopenia syndrome (SFTS), and find the most quick and accurate one for the identification of new bunyavirus infection. Methods Real-time PCR and ELISA-IgM were used to detect serum samples of 158 patients with acute phase of SFTS, which were collected from the special monitoring system of SFTS in Henan Province in 2014. IgM and IgG antibodies were detected by ELISA in 109 acute and convalescent paired serum specimens. The differences of the positive rates were compared between the three methods, and the influence of the collected interval time on the detection results was analyzed. Results For 158 acute phase serum samples of SFTS patients, the positive rate detected by real-time PCR (76.58%) was higher than that of ELISA-IgM (47.47%), and the difference was statistically significant (χ2=34.13, P 0.05). In both the acute phase and convalescent phase, the positive rate of IgM was higher than that of IgG, and the difference was statistically significant (χ2=41.68 and 6.25, P<0.05). With the extension of collected interral time, the positive rates of IgM and IgG antibodies were both increased ( Z=6.42 and 10.08, P < 0.05). Conclusion Real-time PCR is the most sensitive method for the early diagnosis of the SFTS. ELISA-IgG is suitable for the detection of SFTS at recovery period. ELISA-IgM can be used as an assistant method to guide clinical diagnosis.
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<p><b>OBJECTIVE</b>To investigate the etiology of severe hand-foot-mouth disease (HFMD) in children in Henan province.</p><p><b>METHODS</b>A total of 244 HFMD cases admitted to a hospital in Zhengzhou from April to June of 2014 were recruited for research sampling, Real-time RT-PCR, virus isolation, VP1 sequencing and alignment methods were used to test the enterovirus-related etiology. SPSS 17.0 was used in performing statistical analysis.</p><p><b>RESULTS</b>There were 109 severe and 135 mild cases among all the 244 HFMD cases. The number of enterovirus positive stool samples was 229, with positive rate as 93.85%. EV71, Cox A16 and Cox A10 made up 83.84%, 5.68% and 8.30% of the enterovirus etiologicy, strains, respectively. EV71 infection caused 8 HFMD cases with heart-lung failure and 2 death, Cox A10 infection led to 1 HFMD case with heart-lung failure and death. There were statistically differences seen regarding the enterovirus infection rates between severe and the mild HFMD cases (χ(2)=5.312,P=0.021). Statistically significant difference was seen in the constituent ratio of EV71, Cox A16 and the others by Fisher' s exact test (P=0.048). There was statistically significant difference seen between the cardiorespiratory failure rate and the fatality rate by EV71 and Cox A10 infection (χ(2)=0.051,P=0.821; χ(2)=2.198,P=0.138). Cox A10 strains idenfied in Henan in 2014 belonged to genotype 6. The rates on homology of nucleotide and amino acid among the Cox A10 strains in Henan in 2014 were 94.3%-99.7% and 96.3%-100.0% respectively.</p><p><b>CONCLUSIONS</b>EV71 still remained the most common pathogen that causing severe HFMD in children, with the increasing Cox A10 percentage in the pathogens spectrum of HFMD infection. Cox A10 strains in Henan in 2014 belonged to genotype 6. Genotype 6 Cox A10 had appeared and widely distributed in Henan for long time, but not yet variated or reconstructed. Cox A10 infection could lead to cardio-respiratory failure thus called for the monitoring program on non-EV71 and non-Cox A16 enterovirus, especially Cox A10 to be strenthened.</p>
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Child , Humans , Amino Acids , Genetics , Biometry , Enterovirus A, Human , Classification , Genetics , Enterovirus Infections , Epidemiology , Virology , Evolution, Molecular , Genotype , Hand, Foot and Mouth Disease , Epidemiology , Virology , Hospitals , Real-Time Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To diagnose imported dengue fever case from Henan province, and to sequence and analyze the characteristics of whole genome sequence, and to explore the possible viral origin source.</p><p><b>METHODS</b>A suspected dengue fever case was reported in Yuzhou city, Henan province. The patient returned from foshan, Guangdong province on September 19, 2014, after the epidemiological investigation and serum specimen collected, which dengue fever case was diagnosed in the laboratory, then it was inoculated on Vero cells. Whole genome sequence was amplified by several pairs primers and characterized using biologic software.</p><p><b>RESULTS</b>The imported case was diagnosed as dengue virus 1 serotype infection. Dengue 1 strain was isolated using Vero cells successfully. Whole genome was 10,670 nt, which belonged to dengue virus 1 serotype V genotype and didn't found any recombination event. The phylogenetic analysis demonstrated that the strain was closed to Indian starins isolated in 2008-2011, and the homology of nucleotide sequence was between 98.2%-99.4%.</p><p><b>CONCLUSION</b>It was the first time to discover imported dengue 1 serotype case in Henan province. However, according to the patient has been to Guangdong province before onset, it inferred that the Indian strain had been imported to Guangdong province before this case in Henan province.</p>
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Animals , Humans , Chlorocebus aethiops , China , Dengue , Dengue Virus , Genes, Viral , Genotype , India , Molecular Epidemiology , Serogroup , Vero CellsABSTRACT
[ ABSTRACT] AIM:To compare the expression of SIRT2 in ovarian surface epithelial ( OSE) cell line and serous ovarian carcinoma ( SOC) cell lines, and to investigate the effects of SIRT2 on the cell proliferation, migration and inva-sion.METHODS:The expression levels of SIRT2 in the OSE cell line and the SOC cell lines were determined by Western blot.The SIRT2 siRNAs and overexpression construct were designed and verified.Transient transfection of SIRT2 siRNAs or overexpression construct was performed, and the effect of SIRT2 on the cell proliferation, migration and invasion was e-valuated.RESULTS:SIRT2 levels in the 5 strains of SOC cell lines were significantly lower than that in the OSE cell line.SIRT2 knockdown in HOSEpiC cells significantly enhanced the ability of cell colony formation and accelerated the cell growth rate.On the contrary, overexpression of SIRT2 in HO8910 cells dramatically repressed the number of cell colonies and cell activity.SIRT2 significantly changed the ability of ovarian cell migration.Knockdown of SIRT2 facilitated the cell invasion.CONCLUSION:The expression of SIRT2 in the SOC cells is significantly down-regulated.In the OSE cells, SIRT2 acts as a tumor suppressor and mediates the inhibition of cell proliferation, migration and invasion.
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<p><b>OBJECTIVE</b>To clone and express the recombinant capsid protein VP2 of enterovirus type 71 (EV71) and to identify the immune activity of expressed protein in order to build a basis for the investigation work of vaccine and diagnostic antigen.</p><p><b>METHODS</b>VP2 gene of EV71 was amplified by PCR, and then was cut by restriction enzyme and inserted into expression vector pMAL-c2X. The positive recombinants were transferred into E.coli TB1, the genetically engineered bacteria including pMAL-c2X-VP2 plasmids were induced by isopropyl thiogalactoside ( IPTG) , and the expression products were analyzed by SDS-PAGE and western blotting method. EV71 IgM antibody detection method by ELISA was set up, and the sensitivity and specificity of this method was assessed; 60 neutralizing antibody positive serum samples from hand foot and mouth disease (HFMD) patients were determined, of which 52 samples were positive and 8 samples were negative; a total of 88 acute phase serum samples of HFMD patients diagnosed in clinical were also detected.</p><p><b>RESULTS</b>VP2 gene of 762 bp was obtained by PCR, the gene segment inserted into the recombinant vector was identified using restriction enzyme digestion. The recombinant vector could express a specific about 71 500 fusion protein in E.coli by SDS-PAGE. The purified recombinant protein of EV71-VP2 can react with the serum of HFMD patients to produce a specific band by western blotting. The sensitivity and specificity of ELISA was 87% and 83%, respectively. Of the 88 acute phase serum samples from children with HFMD, 48 samples (55%) were positive by the ELISA assay.</p><p><b>CONCLUSIONS</b>VP2 gene of EV71 has been cloned and a prokaryotic high expression system for VP2 gene was successfully constructed in the present study. The recombination EV71-VP2 has well antigenicity, which could be useful for developing diagnose reagent or vaccine of EV71.</p>
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Humans , Antibodies, Neutralizing , Blood , Antibodies, Viral , Blood , Capsid Proteins , Genetics , Allergy and Immunology , Enterovirus A, Human , Genetics , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Genetic Vectors , Hand, Foot and Mouth Disease , Allergy and Immunology , Immunoglobulin M , Blood , Recombinant Proteins , Genetics , Allergy and ImmunologyABSTRACT
To investigate the animals infection situation of novel bunyavirus in Xinyang City ,Henan Province ,China , animal serum samples such as cattle ,dog ,swine ,mice were collected in Shangcheng County and Guangshan County in Xinyang City .All the serum samples were detected by novel bunyavirus ELISA and real time RT-PCR method .A total of 292 animal serum samples were collected including 5 kinds of animals .The result of all the animal serum samples were negative by using real time RT-PCR ,and the positive rate was 45 .19% (141/312) by ELISA method .Of the 5 animal serum samples including mice ,cattle ,goats ,swine and dogs ,the positive rate were detected to be 1 .06% ,100 .00% ,76 .27% ,3 .57% ,and 75 .00%respectively .There was significant difference in results among 5 kind of animal serum antibodies .Animals such as cattle and dog may be the host of novel bunyavirus which were detected novel bunyavirus antibodies in cattle and dog in Xinyang City , Henan Province ,China .
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Objective:To elucidate the clinical effect of xingxiong sodium chloride injection on vertebralbasilar insufficiency.Method:98 patients with vertebralbasilar nsufficiency were randomly divided into two groups,(therapy group with 50 cases treated with xingxiong sodium chloride injection and the controlled group with 48 treated with danshen injec- tion.After 14 days,their clinical symptom change and haemodynamics parameters change were evaluated and detected by transcranial Doppler (TCD).Result:In the therapy group,45 cases (90%) were cured and in the controlled,30 cases (62.5%) were cured.The two groups presented a statistical significance (P