Objective To analyze the evolutionary characteristics and predict the variation trend of neuraminidase (NA) gene of influenza A/H1N1 (09pdm) virus in China from 2009 to 2016 in order to provide the basis of assessment for flu vaccines.Methods A total of 1 141 sequences of NA gene of influenza A/H1N1 (09pdm) virus were screened out from the Global Initiative on Sharing All Influenza Data and National Center for Biotechnology Information.The phylogenetic trees and the mutations of amino acids sequences were constructed and analyzed by biological softwares.The prediction for epidemic trend of influenza was analyzed by Bayesian skyline Plot.Results Compared with the sequence of reference strain,the homology of nucleic acid sequence of NA gene decreased year by year from 2009 to 2016.The phylogenetic analysis showed that NA gene clustered nearly on the identical phylogenetic tree in one year.The positive selection pressure site of NA strain was observed by different models in each year except 2012.The dynamics analysis showed that the popularity of influenza A/H1N1 virus may continue to increase to a peak in 2017.Conclusion The amino acid encoded by NA gene of influenza A/H1N1 virus is varying gradually,so the importance of surveillance for influenza virus should be reinforced for every year.

Objective To understand the correlations between HIV-1 subtypes and changes in CD4+T cell count over time in patients with different subtypes of HIV-1 infection.Methods A total of 94 patients who were diagnosed with HIV-1 infection in Nanjing and received at least twice CD4+T cell counting test before antiretroviral therapy (ART) were recruited in this study.Descriptive analysis was used to present the rates of CD4+T cell decline for different HIV-1 subtypes.Logistic regression analysis and nonparametric test were conducted to investigate the factors responsible for CD4+T cell decline and to analyze the correlations between the rates of CD4+T cell decline and HIV-1 subtypes.Results The median monthly rate of CD4+T cell decline was-2.20 [interquartile range (IQR):-11.36-2.13] cell/μl.Of all patients,25.5% (24/94) had a significant decline (≥30%) in CD4+T cell count.Compared with the patients infected with CRF01_AE,those infected with CRF07_BC (OR=0.28,95%CI: 1.7-6.5) or other subtypes (OR=0.16,95%CI: 1.0-2.9) had a lower risk of significant decline in CD4+T cell count.In addition,results of the nonparametric test showed that the patients infected with CRF01_AE (M=-21.54,IQR:-30.97——11.92 cell/μl) had a faster CD4+T cell loss than those infected with CRF07_BC (M=-11.26,IQR:-14.06——5.63 cell/μl) (P=0.033).Conclusion HIV-1 subtype is associated with the rate of CD4+T cell decline.It is important to monitor the changes in CD4+T cell count in patients infected with CRF01_AE and to carry out timely ART.

Objective To investigate the diagnostic value and significance of myocardial injury markers in neonates.Methods A retrospective analysis of 838 cases of myocardial injury markers in neonates,the total positive rate of hs -cTnI and the positive rates of hs -cTnI(hypersensitivity cardiac troponin I)in different kinds of neonatal disease were calculated,the levels of abnormalities consistent rate in hs -cTnI and CK -MBmass(creatine kinase MB mass)were compared with hs -cTnI and MYO(Myoglobin).Results The total positive rate of hs -cTnI was 40.33% in the 838 neonates.The highest incidence of myocardial injury was neonatal sepsis (57.14%),followed by neonatal pulmonary hemorrhage (55.56%)and neonatal convulsions (54.55%).The abnormalities consistent rate of hs -cTnI and CK -MBmass was better than hs -cTnI and MYO (85.50% vs 28.11%,χ2 =226.9,P <0.05). Conclusion Neonatal hospitalized children often complicated by myocardial injury;As a biochemical myocardial injury marker,the hs -cTnI detection is important for early detection of myocardial injury,it should be recommend as routine test items;CK -MBmass has better correlation with hs -cTnI than MYO,which can provide guide for doctors to interpret the data of myocardial injury markers.

BACKGROUND:Large numbers of experimental data have confirmed that bone marrow mesenchymal stem cel s have a positive therapeutic effect on cerebral ischemia-reperfusion injury, but there are few reports about intravenous administration of bone marrow mesenchymal stem cel conditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the effects of the conditioned medium of rat bone marrow mesenchymal stem cel s on the recovery of neurological function in rats after cerebral ischemia-reperfusion injury.
METHODS:The bone marrow mesenchymal stem cel s were isolated from rat bone marrow. When cel s at passage 2 or 3 reached 90%confluence, the original culture medium was removed. Then the cel s were cultured in serum-free DMEM for 18 hours. After that, the culture solution was col ected as the conditioned medium of rat bone marrow mesenchymal stem cel s. Adult rats were subjected to 2 hours of right middle cerebral artery occlusion. Ischemia-reperfusion injury rats were randomly assigned to three groups:control group, simple culture medium group and conditioned medium group, and respectively given injection of normal saline, DMEM, conditioned medium (10 mL/kg) via the tail vein at 2, 24, 48 hours after operation.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperative 2 hours (P>0.05). Compared with the control and simple culture medium group, neurological impairment was significantly improved in the conditioned medium group at postoperative 1, 3, 5 days (P<0.05), but there was no significant difference between the control and simple culture medium groups. At postoperative 5 days, brain edema was significantly eased in the conditioned medium group in comparison to the control and simple culture medium groups (P<0.05), and there was also no difference between the latter two groups (P>0.05). These results suggest that rat bone marrow mesenchymal stem cel s-conditioned medium via intravenous administration can significantly ease brain edema and improve the neurologic function after cerebral ischemia-reperfusion injury.

OBJECTIVETo understand if the Neuraminidase (N1) of Influenza A virus at the surface of yeast-displaying system, eukaryotic expression system and the infected cells could be used for anti-NA Abs screening, their activities and bindings to five candidate Abs were assayed.

METHODSThe surface NA expression was obtained by transfecting by recombinant NA constructors with specific tag-labels or live virus infection. The functional activity was measured by the fluorescent assay. Their bindings to the Abs were detected by flow cytometry.

RESULTSThe surface NAs presenting on the yeast-displaying system and eukaryotic expression system exhibited functional NA activities as the NA at the surface of virus-infected cells which showed affinities to Ab1, 4, and 5. The same bindings to Abl and 5 were found in the surface NA expressed by eukaryotic expression system while minor binding was observed in the yeast displayed-NA.

CONCLUSIONThe epitopes of yeast-displayed NA may be different from the NAs present at eukaryotic expression system and the infected cells which more likely suitable for the screening of anti-NA Abs.

Antibodies ; immunology ; Antigens, Surface ; genetics ; immunology ; Cell Line ; HEK293 Cells ; Humans ; Influenza A virus ; genetics ; immunology ; Neuraminidase ; genetics ; immunology ; Protein Binding ; Recombinant Proteins ; genetics ; immunology

Objective To construct the molecular transmission networks of HIV-1 CRF01 AE strains circulating in China and to analyze their characteristics. Methods Sequences of the pol genes of Chinese CRF01 AE strains were downloaded from Los Alamos database and the related demographic informa-tion was checked. Transmission networks were created by four steps,including phylogenetic tree construc-tion,transmission cluster extraction,minimum genetic distance identification and network visualization. Chi-square test was performed to analyze the differences in the distribution of different populations in the networks and the differences in the distribution of subjects with different degrees among different sub-populations. De-scriptive analysis was used to investigate the transmission links between sub-populations and various regions. Results The 2 419 sequences constituted 250 separate networks including 847 nodes and 610 edges. The number of subjects covered by different networks ranged from 2 to 25. Subjects with degree ≥2 represented only 26. 4%(224 / 847)of network-individuals,but were connected with 66. 5%(563 / 847)of all network-individuals. There were 37. 6%(669 / 1 781)of men who have sex with men(MSM),28. 4%(102 / 359) of heterosexual men,16. 1%(10 / 62)of intravenous drug users(IDUs)and 30. 4%(66 / 217)of hetero-sexual women involved in the networks(χ2 =23. 774,P﹤0. 001). The percentage of subjects with degree ≥2 was high in MSM(10. 8% ,193 / 1 781)compared to that in heterosexual men(5. 6% ,20 / 359)(P =0. 002)and heterosexual women(4. 6% ,10 / 217)(P=0. 004). Of the 669 MSM in the networks,95. 5%(639 / 669)linked to other MSM and only 2. 4% (16 / 669)linked to heterosexual women. However, 15. 1%(10 / 66)of the heterosexual women in the networks linked to MSM. Of the heterosexual men in the networks,35. 3%(36 / 102)linked to heterosexual women,9. 8%(10 / 102)linked to MSM. Of the sub-jects in the networks,20. 9%(177 / 847)linked to other regions' individuals. Conclusion The super-spreaders played an important role in the molecular transmission networks of HIV-1 CRF01 AE strains even though they were in a minority. The transmission of HIV-1 CRF01 AE strains between sub-populations and various regions was complicated and active.

OBJECTIVESTo study the inhibitory effect of specially synthesized oligodeoxynucleotide (SODN) on malignant phenotype of human hepatocellular carcinoma Hep3B cells by complementary binding to the fourth promoter of IGF-2 gene.

METHODSA SODN was synthesized according to the sequence of the fourth promoter of IGF-2 gene, and was then transfected into Hep3B cells which overexpressed IGF-2. IGF-2 gene transcription activity and protein expression were assayed by RT-PCR and Western blot methods. The effect on cell growth by SODN was estimated by MTT method, and the effect on cell cycle distribution was measured by flow cytometry. Colony formation assay was performed on 6-well tissue culture dishes. Alteration on mobility and invasiveness were studied used transwell plates.

RESULTSIGF-2 mRNA and protein levels in Hep3B cells transfected with SODN were significantly lower in comparison with those in control groups (Hep3B cells and Hep3B cells transfected with a control oligodeoxynucleotide). Results also showed that SODN did not have inhibitory effects on cell growth and mobility of Hep3B cells, but did have an effect on its colony formation and invasiveness.

CONCLUSIONSODN has inhibitory effect on IGF-2 expression in Hep3B cells as a molecular switch, which partially alterates the malignant phenotype of this cell line.

Carcinoma, Hepatocellular ; pathology ; Humans ; Insulin-Like Growth Factor II ; biosynthesis ; genetics ; Liver Neoplasms ; pathology ; Neoplasm Proteins ; biosynthesis ; genetics ; pharmacology ; Oligonucleotides, Antisense ; biosynthesis ; genetics ; pharmacology ; Phenotype ; Tumor Cells, Cultured

OBJECTIVETo understand the allele structure and genetic polymorphism at F13A01, FESFPS and vWA short tandem repeats (STRs) loci for Han population in Guizhou province of China, and to construct a preliminary database.

METHODSOne hundred sixty unrelated individuals in Han population from Guizhou were analyzed by multiplex polymerase chain reaction. The resultant allele frequencies were compared with those of other population or race database.

RESULTSThe genotype distributions of F13A01, FESFPS and vWA were in accordance with Hardy-Weinberg equilibrium. The combined discrimination power and polymorphism information content were 0.9984 and 0.9700 respectively.

CONCLUSIONAll of the three loci in this study provide useful markers for forensic paternity test and for genetic purposes.

Alleles ; China ; Gene Frequency ; Humans ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic

This article reports two cases of childhood-onset nemaline myopathy diagnosed by muscle pathology and genetic diagnosis. The two patients had onset in early childhood, with muscle weakness as the first manifestation, as well as long disease duration and slow progression. Gomori staining and hematoxylin-eosin staining showed red-stained rods in the sarcoplasmic cytoplasm and sarcolemma under a light microscope. Electron microscopy showed that the dense nemaline rods were located under the muscle fiber sarcolemma and parallel to the long axis of the muscle fibers, and some muscle fiber myofilaments were dissolved and necrotic. Gene testing found that one of the two patients had heterozygous mutation (c.1013A>C) in the ACTA1 gene, and the other had compound heterozygous mutation (c.18676C>T and c.9812C>A) in the NEB gene. The two mutations were more common in nemaline myopathy. Nemaline myopathy is a recessive or dominant inheritance myopathy, in which the nemaline rod in the cytoplasm of myocytes is a characteristic muscle pathological change. Pathological and genetic diagnosis is the gold standard for diagnosis of nemaline myopathy.

Objective To investigate the expression of thymidine kinase 1 (TK1) in benign and malignant tumors.Methods From Jan.2013 to May.2015,180 female patients with breast diseases aging 16 to 79 years,including 112 cases of breast cancer (breast cancer group),and 68 cases of breast fibroadenoma (breast fibroadenoma group) were collected.The mean age of the patients was 47 years,with a median age of 43 years.We collected all the patients' fasting venous blood,detected the level of TK1,compared their differences of expression levels between patients with fibroadenoma and breast cancer,and the differences in serum TK1 levels in patients with different pathological characteristics.Results The level of TK1 in breast cancer patients was significantly higher than that in patients with fibroadenoma,and the difference was statistically significant (P＜0.01).In breast cancer group,the tumor size was not correlated with TK1 expression of (P＞0.05).The level of serum TK1 in patients with advanced TNM stage was higher than that in patients with early TNM stage (P=0.038).The level of serum TK1 in patients with advanced lymph node stage was higher than that in early lymph node stage (P=0.048).The level of TK1 in M1 patients was higher than that in M0 patients and the difference had statistical significance (P=0.018).Conclusion TK1,as a new marker of cell proliferation index,can accurately reflect the proliferation of tumor cells in human body,which can be used as an important index to monitor the degree of tumor proliferation and identify benign and malignant tumors.