Objective To explore the effects of dipeptidyl peptidase ( DPP-4 ) inhibitor on proteins expression of Bcl-2 and Bax of islet β-cells through increasing the expression of islet γ amino acid butyric acid ( GABA) . Methods A total of 50 rats of clean grade were studied. Among them, ten rats were randomly selected as normal controls, the remaining forty rats were fed with high-fat diet and then intraperitoneal injection with streptozotocin, the diabetic rats models were then established. Rats were randomly divided into three groups:i. e. diabetic control group, DPP-4 inhibitor group, and antagonist group ( DPP-4 inhibitor and GABA receptor antagonist). Six weeks later, blood glucose, serum insulin, glucagon, and the proteins expression of GABA, Bcl-2, and Bax of islet β-cells were measured. Results ( 1 ) Compared with diabetic control group, serum insulin was increased(P<0.05),bloodglucoseandserumglucagonweredecreasedinDPP-4inhibitorgroup(P<0.05). (2) Compared with DPP-4 inhibitor group, serum insulin was decreased(P<0. 05), blood glucose and serum glucagon were increased(P<0. 05) in antagonist group. (3) Compared with diabetic control group, the expression of GABA was increased(P<0. 05), the expression of Bcl-2 protein was increased (P<0. 05) in pancreatic β-cells in DPP-4 inhibitor group. ( 4 ) Compared with diabetic control group, the expression of GABA in pancreatic β-cells was increased in antagonist group(P<0. 05) . Compared with DPP-4 inhibitor group, the expression of Bax protein in pancreaticβ-cells was increased in antagonist group(P<0. 05) , while the expression of Bcl-2 protein was decreased (P<0. 05). Conclusions DPP-4 inhibitor could increase the secretion of insulin, decrease the secretion of glucagon, up-regulate expression of anti-apoptosis protein Bcl-2, and down-regulate expression of apoptosis protein Bax in pancreatic β-cells through increasing the expression of GABA, inhibiting pancreatic β-cells apoptosis and protecting the damagedβ-cells in type 2 diabetic rats.

Objective To investigate the effect of infiltrated mast cells on the biological characteristics of uterine carcinoma.Methods Twelve leiomyomas,ten uncertain malignant uterine leiomyomas,and seven uterine leiomyosarcomas were studied with light microscopy for the morphometry about their histological feature,mast cell particle and factor Ⅷ-related antigen(FⅧRAg);Cell apoptosis was measured by flow cytometry.The correlative analysis was carried out between the mast cell and others factors.Results Comparing with smooth muscle tumors of uncertain malignant potential(4209.9?273.0),the count of tumor cell was the highest in leiomyosarcoma(3557.6?346.3)(P

The aim of this study is to analyze the evolutional and molecular characteristics of Hemagglutinin (HA),Neuraminidase(NA)and non-structural (NS) genes of H5N1 avian influenza virus (AIV) from sewage in live bird markets (LBMs) in Changsha,2014.Five hundred and one specimens were collected from environment in LBMs in Changsha,2014,and real-time RT-PCR was used for influenza A typing and subtyping (H5,H7 and H9) detection.Sequencing were used for the positive of single H5.The sequence homology of HA,NA and NS genes of the viruses were analyzed with the online Basic Local Alignment Search Tool (BLAST).The phylogenetic trees for HA,NA and NS genes and the ClustalW Multiple alignments of amino acids were constructed using MEGA 5 and BioEdit software,respectively.Results showed that of 501 environmental samples,177 (35.33 ％) samples were positive for influenza A viruses and H5 subtype.Eight H5N1 subtype AIV were confirmed by sequencing from the samples of the positive of single H5.Phylogenetic analysis indicated that most of HA genes of the H5N1 subtype AIV strains isolated in Changsha city were located in 2.3.2 and clustered into new subclade,and the most of NA and NS genes in this study were clustered into subclade 2.3.2.1b.QSG of the HA protein of the receptor binding site were found in these H5N1 viruses,and the characteristics was shown to be associated with increased affinity of HA to the glycan-receptors of AIV.In strains from this study,we did not found amino acid substitutions of the NA protein at H275Y and N295S,and sensitive to neuraminidases,and the high pathogenicity molecular characteristics of HA,NA and NS genes were showed in these viruses.In conclusion,molecular characteristics of the HA,NA and NS of these H5N1 subtype viruses in this study showed high pathogenicity,but that may not facilitate human infection.So,the prevalence and genetic evolution of this virus should be closely monitored.

Objective:To investigate the correlations of Lauren classification and world health organization (WHO) classification of gastric cancer (GC) with microvascular density (MVD), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2), and p53. Methods:The clinical data of 89 patients with GC were collected. The collected specimens were categorized on the basis of Lauren classification and WHO classification. CD34/periodic acid-Schiff (PAS) double staining was performed to validate MVD. Immunohistochemistry was conducted to investigate the expression levels of MMP-2, MMP-9, VEGF, VEGFR1, VEGFR2, and p53. Results:MVD was not correlated with Lauren classification or WHO classification (P>0.05). Lauren typing was associated with the expression levels of MMP-9, VEGFR1, and p53 (P<0.05). WHO classification was not related to any of the factors (P>0.05). Cox proportional hazards model revealed that Lauren classification and WHO classification were the prognostic factors of overall survival (P<0.05). Conclusion:This research on tumor related factors, angiogenesis, and different classifications of GC may provide new methods to treat this disease.

We analyzed the evolutional and molecular characteristics of Hemagglutinin(HA),Neuraminidase(NA) and nonstructural(NS) genes of avian influenza A(H9n2) viruses from environment in poultry markets in Changsha,China,2014,providing laboratory data for prevention and control of human infection with avian influenza A(H9N2) virus.Five hundred and one specimens (263 poultry drinking water specimens,226 poultry sewage specimens and 17 others specimens) were collected from environment in poultry markets in Changsha,2014,and real-time RTPCR was used for influenza A typing and subtyping (H5,H7 and H9) detection.HA and NA universal primer sets for conventional RT-PCR and sequencing were used for the positivity of single H9.The sequence homology of HA,NA and NS genes of the viruses were analyzed with the online Basic Local Alignment Search Tool (BLAST).The ClustalW multiple alignments of amino acids and the phylogenetic trees for HA,NA and NS genes were constructed using the BioEdit and MEGA 5 software,respectively.Results showed that among 501 environmental samples,350 samples were positive for influenza A virus,191 (38.12％) for H9 subtype,177 (35.33％) for H5 subtype,11 (2.20％) for H7 subtype and 68 (13.57％) for H5 and H9 subtypes co-detection.Twenty-three H9N2 subtype AIV were confirmed by conventional RT-PCR and sequencing from the samples of the positivity of single H9.Phylogenetic analysis revealed that most of HA,NA and NS genes of the H9N2 subtype AIV isolated in Changsha City had gene constellations of genotype S,and these virues might have acquired their HA,NA and NS from A/Chicken/Shanghai/F/1998-like (H9N2).L235 (correspond to H3 numbering 226) of the HA protein of the receptor binding site (RBS) were found in these H9N2 viruses,and the characteristics was shown to be associated with increased affinity of HA to the glycan-receptors of human influenza virus,and the low pathogenicity molecular characteristics of HA,NA and NS genes were showed in these viruses.The positive rate of nucleic acid of the H9 subtype of avian influenza virus from environment was the highest in poultry markets in Changsha,2014,and molecular characteristics of the HA,NA and NS of these H9N2 subtype AIV showed low pathogenicity,but that may facilitate human infection.So,the prevalence and genetic evolution of this virus should be closely monitored.

BACKGROUNDConstructing replication defective recombinant adenovirus vector expressing the group specific antigen VP6 of human rotavirus and studying the immune responses induced in vivo.

METHODSThe cDNA of full length VP6 was inserted into the adenovirus vector pShuttle-CMV, and recombinant adenovirus genome DNA was obtained through homological recombination in E.coli,then the recombinant adenovirus was gained after transfecting 293 cell line with the genome DNA. Gene integration of VP6 in resultant adenovirus was confirmed by PCR and Southern blot, respectively gene expression was confirmed in 293 cells by Western blot. BALB/c mice were immunized intranasally(inl)and orally(ora), respectively, to test the immunization effects of the adenovirus.

RESULTSRecombinant adenovirus named rvAd-VP6 was obtained. The cDNA of VP6 was integrated in the adenovirus and was able to be expressed in 293 cells stably. The systemic immune responses to rotavirus VP6 could be induced effectively in both oral and intranasal group, the titer of serum IgG antibody in the two group of mice were 1?1 000 and 1?10 000-1?100 000, respectively. In addition to IgG, the serum IgA specific to VP6 could also be detected at a titer of 1?10-1?100. Secretory IgA(sIgA) was detected in both lung lavage fluid and intestinal homogenate when administered intranasally to BALB/c mice, whereas only found in intestinal homogenate in the oral group. The results indicated that the immunization efficacy of intranasal inoculation was superior to that of oral inoculation.

CONCLUSIONSThe recombinant adenovirus vector expressing human rotavirus VP6 was successfully constructed, its ability to induce immune responses has laid a solid foundation for the development of rotavirus genetically engineering vaccine against rotavirus infection.

Adenoviridae ; genetics ; Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; Capsid Proteins ; biosynthesis ; immunology ; Genetic Vectors ; Mice ; Mice, Inbred BALB C ; Recombination, Genetic ; Rotavirus ; immunology

BACKGROUNDTo express and purify human alpha3-integrin to serve as the antigen to prepare its antibody and to separate the Vero cell clones without expression of alpha3-integrin.

METHODSThe human alpha3-integrin gene was amplified by using RT-PCR, then subcloned into a pQE30 expression vector and expressed in E. coli. The gene expression was confirmed by Western blot assay. Rabbit was inoculated with purified antigen to stimulate the antibody generation. The target Vero cells were separated by negative selection using antibody plus complement mediated cytolysis. The separated cell clones were confirmed by immunofluorescence and Western blot assay.

RESULTSThe alpha3- integrin gene was cloned and expressed effetively, Western blot assay revealed that the expressed protein held good immune reactivity. High titer antibody was generated. However the expression of alpha3-integrin was not detected on Vero, VeroE6, Hep-2, 2BS and 293 cells.

CONCLUSIONSThe results of the study suggested that hantavirus has other receptors on Vero cells beside alpha 3-integrin.

Animals ; Cercopithecus aethiops ; Cloning, Molecular ; Gene Expression ; Hantavirus ; Integrin beta3 ; biosynthesis ; genetics ; immunology ; Rabbits ; Receptors, Virus ; Vero Cells ; metabolism

Autoimmune encephalitis (AE) is a kind of encephalitis mediated by autoimmune mechanism. Cognitive impairment is one of the main manifestations of AE at acute phase. Cognitive impairment is also found during long-term follow-up in some AE patients who have good prognosis after immunotherapy or tumor resection. In addition, the proportion of patients with long-term cognitive impairment, main cognitive impairment domains and risk factors are different in various types of AE. This paper summarizes the cognitive impairment in different types of AE, hoping to improve the clinicians' understanding of cognitive impairment in AE.

Objective:To carry out the monitoring of H5N6 subtype avian influenza virus (AIV) in the population infection and live poultry markets (LPMs) environmental pollution in Changsha city from 2016 to 2018. Laboratory data were provided to prevent human infection with H5N6 subtype AIV.Methods:A total of 6 909 throat swab specimens from the population and 1 719 environmental specimens from LPMs were collected in Changsha city from 2016 to 2018. The real-time RT-PCR method was used for the detection of influenza virus in type A, H5, H7, H9 and N6 subtypes. Eighty-two AIV nucleic acid-positive specimens were subjected to high-throughput nucleotide sequencing, and then BLAST similarity and amino acid (aa) key site analyses were performed on the sequencing results.Results:One H5N6 subtype AIV nucleic acid was detected from 6 909 population throat swab specimens, 927 (53.93%) positive specimens of A-type AIV nucleic acids and 193 (11.23%) positive specimens of both H5 and N6 subtypes were detected from 1 719 LPMs environmental specimens. Fourteen genomic sequences of H5N6 subtype AIV were obtained with high-throughput nucleotide sequencing, and aa key site analyses showed that there were 6 basic aa in 338-347 aa of viral HA protein-linked peptide, which was highly pathogenic to birds. Receptor binding site (RBS) 238-240 aa (corresponding to H3 type influenza virus 226-228 aa) is QSG or QRG, and the receptor is characterized by avian origin. The 290th resistance gene locus of NA protein did not show R290K mutation, and was sensitive to NA inhibitor (Duffy/osoterosine phosphate). The E627K and D701N(I) mutations in the viral PB2 protein indicate that the virus is pathogenic.Conclusions:Human infected with H5N6 subtype AIV in Changsha city is sporadic, and the H5N6 subtype AIV of LPMs is seriously polluted. It is necessary to further strengthen the AIV surveillance of LPMs.

Objective:To evaluate the practical value of ultrasound, ultrasound new technologies (ultrasound elastography, contrast enhanced ultrasound) and computed tomography (CT) examination alone or the combination of both of the four in the clinical diagnosis of lymph node metastasis of thyroid cancer by meta-analysis system.Methods:This study screened relevant literature on ultrasound, new ultrasound technologies, and CT in the diagnosis of lymph node metastasis of thyroid cancer by searching Chinese and English databases from January 2006 to August 2023. Two scholars independently assessed the quality of the literatures and scored them using the QUADAS scale. Then, the diagnostic ratio of single and combined examinations was analyzed by Meta-Disc software, and the heterogeneity test and meta-analysis were performed to calculate the sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic ratio, accuracy, and area under curve (AUC). Finally, the diagnostic efficacies of different diagnostic techniques and their combination in identifying lymph node metastasis of thyroid cancer were assessed by calculating OR and P values by two-by-two comparisons. Results:Sixty-nine diagnostic studies totaling 30, 666 lesions were finally included. The meta-analysis study showed that contrast enhanced ultrasound had the highest sensitivity[0.83(0.77-0.87)], ultrasound specificity was the highest [0.84(0.79-0.88)], the highest AUC was for contrast enhanced ultrasound (0.90), and contrast enhanced ultrasound and ultrasound elastography had high accuracies (0.81, 0.80, respectively) both of them were higher than those of ultrasound (0.77) and CT (0.69). Contrast enhanced ultrasound and ultrasound elastography were both more sensitive than CT, with statistically significant differences between them ( P<0.01); contrast enhanced ultrasound + ultrasound elastography had the highest accuracy, while ultrasound + contrast enhanced ultrasound and contrast enhanced ultrasound + ultrasound elastography were both more sensitive, with no statistically significant differences between them ( P>0.05), but the latter had the highest specificity [0.90(0.88-0.92)], which were statistically significantly different from each other ( P<0.05), ultrasound+ contrast enhanced ultrasound and contrast enhanced ultrasound+ ultrasound elastography had higher sensitivities than ultrasound+ CT, which were statistically different from each other ( P<0.05), and ultrasound+ CT had the lowest specificity [0.76(0.70-0.81)]. Conclusions:The diagnostic efficacy of thyroid cancer lymph node metastasis is highest with contrast enhanced ultrasound. The new ultrasound technique for diagnosing thyroid cancer lymph node metastasis has a higher diagnostic value than CT, and it can be used as a complementary examination to conventional ultrasound.When the two techniques are used in combination, contrast enhanced ultrasound + ultrasound elastography has the highest accuracy, contrast enhanced ultrasound + ultrasound elastography and ultrasound + contrast enhanced ultrasound both have high sensitivity and specificity, and the diagnostic performance of both is higher than that of ultrasound + ultrasound elastography and ultrasound + CT, and in actual clinical practice when ultrasound elastography combined with contrast enhanced ultrasound is limited in the examination, ultrasound combined with contrast enhanced ultrasound can effectively serve as an alternative means to provide a good diagnostic support.