Isolation and genome-wide evolutionary analysis of a type Ⅲ WU polyomavirus strain / 中华微生物学和免疫学杂志

Objective:To isolate and culture WU polyomavirus (WUPyV), and to analyze the genome-wide evolutionary patterns, homology and population dynamics.Methods:Real-time quantitative PCR was used to detect the nasopharyngeal aspirate samples of hospitalized children with respiratory tract infection in Beijing Friendship Hospital during 2020 to 2022. Primary human airway epithelial cells cultured at the air-liquid interface were used to isolate and culture WUPyV. Whole genome sequence of the isolated strain was obtained by Sanger sequencing. For phylogenetic and evolutionary dynamics analysis, the whole genome was compared with the published whole genome sequences in GenBank database.Results:The detection rate of WUPyV was 4.7% (31/659) during 2020 to 2022, and a clinical strain BJ0593 of WUPyV type Ⅲc was successfully isolated. The homology of the whole genome and gene fragments of WUPyV was high. The average evolutionary rate of VP2 gene was about 1.256×10 -4 substitution/site every year, and the population dynamics of WUPyV tended to be flat in the last decade. Conclusions:This study successfully isolated a clinical WUPyV type Ⅲ strain for the first time, which provided the basis for further investigation on the molecular evolution and pathogenicity of WUPyV.

Internet addiction and college students depression:an analysis of moderating mediating effects / 中国学校卫生

Objective@#To explore the mediating role played by college students social anxiety and social support in the relationship between internet addiction and depression.@*Methods@#A cluster random sampling method was used to conduct a questionnaire survey among 3 536 college students in three higher vocational colleges in Anhui Province. The content included general demographic characteristics, depression, Internet addiction, social support, and social anxiety. The Process program was used to mediate and analysis of regulation.@*Results@#Among the survey subjects, 1 552(43.90%) had depressive symptoms, including 561(45.65%) boys and 991(42.96%) girls.The total score of Internet addiction was significantly positively correlated with depression score(r=0.30, P<0.01); social anxiety(social fear, social avoidance) was positively correlated with depression(r=0.24, 0.27, P<0.01); social support(subjective support, objective support, support utilization) was significantly negatively correlated with depression(r=0.25, -0.23, -0.17, P<0.01). Conditional process analysis shows that social anxiety had a mediating role between internet addiction and depression(c'=0.06, P<0.01), and that Internet addiction and social anxiety were regulated by social support(β=-0.00，P=0.02).@*Conclusion@#By increasing the social support of college students to improve social anxiety, it might help to reduce the depression of college students caused by Internet addiction.

Correlation and modulation of smooth muscle cells phenotypic switching induced by cytomegalovirus / 中华实验和临床病毒学杂志

Objective@#To explore the influence of murine cytomegalovirus on phenotypic modulation of vascular smooth muscle cell and modulation of PI3K/Akt pathway.@*Methods@#Male apoE knockout mice were injected abdominally with 2×105 PFU MCMV, followed by 16 weeks feeding. Then aortas were sectioned for HE staining and immunohistochemical staining of smooth muscle 22 alpha (SM22α) and osteopontin (OPN). Mouse aortic smooth muscle cells(MOVAS)were incubated with MCMV, then proliferation of MOVAS and expression of SM22a and OPN were tested. Western blotting test was applied to reveal MCMV’s modulation of PI3K/Akt pathway.@*Results@#The degree of atherosclerosis of apoE-/- mice in MCMV infection group was severe than that in control group, and OPN stain positive signals predominated in the arterial wall. After 24 hours of incubation with MCMV by 3×104 PFU, the expression of SM22a decreased (P=0.023), while OPN increased (P=0.034) in MOVAS. MCMV increased expression of Akt phosphorylation compared with the control group (P=0.035). The inhibitor of PI3K pathway LY294002 not only inhibited the phenotypic modulation of smooth muscle by MCMV, but also blocked the Akt phosphorylation after MCMV infection (P=0.031), however no significant influence was observed in control group.@*Conclusions@#MCMV induces phenotypic modulation of vascular smooth muscle, PI3K/Akt signaling pathway may be involved in the process of MCMV promoting the phenotype transformation of smooth muscle cells.

Correlation analysis between ALT and viral hepatitis in blood screening among blood donors / 国际检验医学杂志

Objective To study the correlation between alanie aminotransferase(ALT) unqualified samples and hepatitis B sur‐face antigen(HBsAg) and hepatitis C virus antibody (anti‐HCV) detection and to investigate an effective measure for reducing the discard rate of donated blood .Methods 330 633 blood samples donated by volunteers in Shenzhen Municipal Blood Center from January 1 ,2009 to December 31 ,2013 were performed the ALT ,HBsAg and anti‐HCV detection .Then the correlation between the detection results of ALT and viral hepatitis .Results Among 33 0633 donated blood samples ,there were 932 cases (0 .282% ) of ALT positive and 2 965 cases (0 .897% ) of viral hepatitis positive ,the differences were statistically significant (P<0 .05) .915 cases were unqualified in ALT ,but negative in viral hepatitis ,which accounting for 98 .176% of all ALT unqualified samples ;the blood discard rate generated by ALT disqualification was 0 .277% (915/330633) .Conclusion Our study indicates that the statistical difference exists in the ALT unqualified rate and the viral hepatitis detection rate ,conducting the ALT detection has the lower coin‐cidence rate for expected viral hepatic ,many false positive lead to the discard of normal blood .Therefore ,whether to continue using the ALT detection as the auxillary detection indicator is still being negotiated .

The relationship of Bel subgroup and the G952A mutation of the α1,3 galactosyltransferase gene / 中华检验医学杂志

Objective To study the molecular genetic background of Bel subtype at ABO blood group.Methods Three samples and fifteen samples were diagnosed as Bel subgroup and normal control samples by serological test,respectively.The extracted DNA was genotyped by sequence specific primer- polymerase chain reaction foilowed by sequencing for Exon6 and exon7 at ABO locus and clones were sequenced.Results A novel Bel variant allele(GenBank EF117687) was identified in a Bel individual.The Bel allele was different from the regular B101 allele by single 952G>A missense mutation in exon7.resulting in an amino acid subsfitution of Val for Met at 318 locus.No mutations were detected in the fifteen control samples and the other two Bel allele samples.Conclusions The mutation position was fimt found to lie on coding region of ABO gene behind nucleotide 930.The mutation of G952A in the al,3 galactosyhransferase gene may be one of the molecular genetic basis of Bel ohenotype.

Macrolide resistance and molecular mechanisms in Mycoplasma pneumoniae / 中华检验医学杂志

Objective To investigate status of macrolide resistance and determine molecular mechanisms in Mycoplasma pneumoniae.Nethods All of 370 throat swab specimens were cultured to isolate Mycoplasma pneumoniae.Mycoplasma pneumoniae isolates were identified by nested PCR for specific 16SrRNA gene.Antibiotic susceptibility test was done to identify acrolide resistant strains.23SrRNA gene wag amplified by nested PCR followed by direct automatic sequencing method.The DNA sequences were compared to the sequence of Mycoplasma pneumoniae M129(accession no.X68422)to find molecular mechanisms of drug resistance.Results Fifty clinical strains were isolated from 370 specimens.Of 50 strains.4 strains were susceptible to macrulide,46 strains were macrolide resistant with the percentage of 92%.MICs of resistant strains to erythromycin.Azithromycin and josamycin were elevated.The sequence of 23SrRNA gene in 4 Susceptible strains and the reference strain FH was identical to Mycoplagma pneumoniae gene in GenBank.46 resistant strains arbored a point mutation respectively,among them,40 strains had all A to G transition at position 2063.1 strain had an A to C transition at position 2063,the other five strains showed an A to G transition at position 2064.Conclusions Macrolide resistance in Mycoplasma pneumoniae iS very serious health conceru.The point mutation in 23SrRNA.Xpecailly predominant position 2063 mutation contributed to the macrolide resistance in Mycoplagma pneumoniae.The MICs of resistant strains to erythromycin,azithromycin and iosamycin are much higher than Mycoplasma pneumoniae reference strain FH.

Application of short tandem repeat in monitoring engraftment of allo-hematopoietic stem cell transplantation / 中国组织工程研究

BACKGROUND: The judgment of the engraftment of hematopoietic stem cells after transplantation mainly depends on various genetic labeling in vivo, which are different in sensitivity and effectiveness, thus a method with powerful differential ability, high sensitivity and not restricted by sex is to be established.OBJECTIVE: To observe the DNA genetic loci of short tandem repeat in the blood samples of both donors and recipients before allo-hematopoietic stem cell transplantation and those of recipients at different time points after transplantation.DESIGN: An observation measurement.SETTING: Laboratory of Immunogenetics, Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center.PARTICIPANTS: Blood samples of 18 pairs of donors and recipients, who were successfully matched and accepted hematopoietic stem cell transplantation, were selected from the Laboratory of Immunogenetics, Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center from February 2004 to December 2005. Among the 18 patients, there were 10 males and 8 females, with a mean age of 35 years old, including 6 cases of them were donated by relatives with blood relationship, and 12 cases by volunteers without blood relationship. Informed consents were obtained from all the participants.METHODS: The blood samples of both donors and recipients before transplantation and the blood samples of recipients after transplantation were collected, and the fluorescence labeling short tandem repeat technique was used to detect the 15 loci for short tandem repeat and Amelogenin sex locus, so that the differential loci between the donor and recipient could be screened. The engraftment and dynamic changes of the short tandem repeat genes of the donors in the recipients after transplantation were observed, the times for the earliest occurrences of short tandem repeat genes of the donors and the complete chimerism were recorded.MAIN OUTCOME MEASURES: ① Differential genes between the donors and recipients before transplantation;②Times for the earliest occurrences of short tandem repeat genes of the donors and the complete chimerism.RESULTS: All the 18 pairs of donors and recipients were involved in the final analysis of results. Satisfactory results of the typing at the 15 loci for short tandem repeat and 1 sex locus in the 18 pairs of samples of both donors and recipients before transplantation and the sample of the recipients after transplantation respectively. Averagely 12.4 (8-15) differential loci for short tandem repeat could be distinguished between the donors and recipients. ②After transplantation, short tandem repeat genes could be detected the earliest at 8 (5-14) days averagely, It took 14 (9-23) days averagely for short tandem repeat loci to convert from recipient type completely into donor type, and the engraftment converted from the recipient chimerism types completely into the donor types.CONCLUSION: The fluorescence labeling compound amplification of short tandem repeat technique can precisely measure the number of PCR products, describe the engraftment of hematopoietic stem cells and the whole process of development. It can also provide accurate and timely information for the early judgement of engraftment, predicting failure of transplantation and controlling recurrence.

A novel O~1 variant allele at the ABO blood group locus / 中华检验医学杂志

Objective To identify novel ABO allele in Chinese population. Methods The ABO blood group was tested by serological method, and then genotyped by sequence-specific primer (PCR-SSP) , gene cloning and sequence analysis. Results A healthy blood dornor who was diagnosed as having A2 subgroup and A2O1genotype was subjected to ABO gene cloning and sequence analysis. The haplotype-specific sequence analysis indicate that two single-base deletions, where G-deletion at nucleotide position 261 and A-deletion at nucleotide position 496 were determined in the O1 allele. The nucleotide sequence of the novelO1 allele were identical to ABO 0101 allele except for A-deletion at nucleotide position 496 in exon7 of ABO locus. Conclusion We defined this 0 allele as a novel O1 variant allele, and its registered number by GenBank is AY374123.

Study on the FUT2 gene structure of Xinjiang Uighur people of China / 中国输血杂志

Objective To study the Secretor gene (FUT2) molecular structure of Uighur population in Xinjiang area,China. Methods DNA was extracted from 40 Uygur unrelated donors' blood and sequence analysis of FUT2 genes was performed. Results Four mutations in the FUT2 genes of Uighur donors have been identified. The frequencies of mutations were 71.25% for 357T, 28.75% for 357C,77.50% for 385A,22.50% for 385T,70% for 428G,30% for 428A,72.50% for 739G and 27.50% for 739A. Conclusion Based on the characteristics of FUT2 gene structure of Xinjiang Uighur,it cauld be thought that there are some relationships between Xinjiang Uighur, Taiwanese of China and Caucasiany.

Mutational analysis of FUT1 and FUT2 genes in para-Bombay individuals / 中国输血杂志

Objective To study the mutation of FUT1 and FUT2 genes in para-Bombay individual.Methods Direct DNA sequencing of FUT1 and FUT2 gene coding region were analyzed in two individuals with para-Bombay phenotype.Results One individual lost one of the three AG repeats located at nucleotides 547~552 of the FUT1 gene, whereas two of the three T repeats located at nucleotides 880~882 were deleted in the other.Conclusion Two frame-shift mutations of FUT1 gene are responsible for the H antigen deficiency