Objective To identify the point mutation of Cu/Zn superoxide dismutase(SOD1) gene in an amyotrophic lateral sclerosis(ALS) family and observe the value of denaturing high performance liquid chromatography(DHPLC). Methods DHPLC and DNA sequencing were used to examine SOD1 gene of the ALS family which had not been found mutation by PCR-SSCP. Results DHPLC tests proved double peaks in one member(Ⅲ_1), Which indicated the possibility of mutation in SOD1 exon 4. DNA sequencing revealed that there was a heterozygote,with mutation of GAA to GGA in exon 4, and with a substitution of glutacid by glycine. Conclusion As compared with PCR-SSCP, DHPLC technique has proved to be a rapid and reliable method for screening mutation site in large samples.

Objective: To identify the mutation points of Cu/Zn superoxide dismutase(SOD1) gene in an amyotrophic lateral sclerosis(ALS) family with a unique phenotype,and to compare the value of single strand conformation polymorphism(SSCP) and denaturing high performance liquid chromatography(DHPLC). Methods: Five exons of SOD1 gene were amplified by PCR. The difference of these products were analyzed by PCR-SSCP and DHPLC.DNA sequencing was used to examine the mutation. Results: ①Mutations were found in exons 2 and 5 in several family members.DNA sequencing revealed that a base pair insertion occurred in the codon area of exon 2 and in the non-codon area of exon 5.②The results of DHPLC tests proved double peaks in one member with ALS symptoms(Ⅲ1),which indicated the possibility of mutation in SOD1 exon 4.DNA sequencing revealed that there was a heterozygote,with a mutation of GAA to GGA in exon 4 in the member with double peak. Conclusion: ①The mutations in exons 2,4,5 were proved.Insertion of exon 2 may be responsible for the disease of the ALS family in Chongqing.②Compared with PCR-SSCP,DHPLC technique has been proven to be a rapid and reliable method for screening mutation site in large samples.

Objective To investigate the infection status of Mycoplasma pirum among male HIV/AIDS patients in Jiangsu and analyze the risk factors.The genome sequencing of Mycoplasma pirum was completed for the first time.Methods Male HIV infected individuals and AIDS patients confirmed in Jiangsu province were enrolled for 4 repeated cross-sectional studies by means of detecting the first flow urine sample and venous blood sample collected and questionnaire survey after informed consent.Genome sequencing was conducted for Mycoplasma pirum by using Illumina Hiseq 2000 sequencing platform.Results A total of 1 541 HIV/AIDS patients were surveyed in this study.The infection rates of Mycoplasma pirurm was 15.4％.The patients who received no HAART had higher risk to be infected with Mycoplasma pirum (OR=1.344,95％CI:1.008-1.792).Otherwise,high CD4+T counts was a protective factor for Mycoplasma pirum infection (OR=0.600,95％CI:0.444-0.810).Based on the sequencing result,the genome size of Mycoplasma pirum was 850 704 bp,the GC content was 24.21％ the genome contained 708 genes,the total length of genes was 734 085 bp,the average length was 1 037 bp,accounting for 86.29％ of genome.Conclusion More attention should be paid to the high infection rate of Mycoplasma pirum among male HIV/AIDS patients in the future AIDS prevention and control.The first genome sequencing of standard Mycoplasma pirum strain was completed in this study (registering Serial number:AZHZ00000001),which can provide evidence for the further research of gene function and pathogenic mechanism of Mycoplasma pirum.

Objective To understand the distribution of HIV-1 subtype in Wuxi city,to predict the local HIV-1 epidemics.Methods Samples were collected from the'CD4+T lymphocyte monitoring cohort study'in Wuxi from April 2013 to July 2016.HIV-1 gene was amplified,sequenced and with HIV-1 sequence database constructed,using both the ChromasPro 1.6 and MEGA 7.0 softwares.Bayesian phylogenetic inference was used to rebuild the history of HIV-1 transmission,while BEAST 1.7.2 and FastTree 2.1.10 software were used for data analysis.Statistical analysis using SPSS 22.0 software.Results Among the 205 subjects of HIV-1 infection,32.68％(67/205) of them were over 50 years old.Seven subtypes (including CRF01_AE,CRF07_BC,CRF67_01B,B,CRF08_BC,CRF68_0B,CRF78_cpx) and one of the unique recombinant forms (URFs) were detected.The main subtypes were CRF01_ AE (51.67％,93/180) and CRF07_BC (17.22％,31/180).Differences between subtypes and ways of transmission were statistically significant (x2=16.99,P≤0.05).The proportion of CRF67_01B (12.78％,23/180) was higher than before.Results from Bayesian phylogenetic inference analysis showed that the evolution rate was 2.29 × 10-3 and Time to the Most Recent Common Ancestor (tMRCA) was 2 003.10.CRF67_01B was probably related to the reference strains from Jiangsu and Anhui provinces,and had been spreading in Wuxi since 2003.Conclusions HIV-1 subtypes seemed complex and diverse in Wuxi city in 2013-2016,with CRF67_01B being pervasive.Continuous molecular monitor program was still needed to provide reference for the prediction of epidemics,from the molecular perspective.

Objective To explore the value of coronary fat attenuation index(FAI)combined with laboratory indicators in predicting the risk of acute coronary syndrome(ACS)in patients with coronary heart disease(CHD).Methods A retrospective analysis was conducted on 454 patients who were diagnosed with CHD in Guangdong Provincial People's Hospital SCAD group(n=233)and an ACS group(n=221).Univariate and multivariate Logistic regression analyses were performed on the FAI values of the main coronary branches[right coronary artery(RCA),left anterior descending branch(LAD),left circumflex branch(LCX)],laboratory indicators,and clinical data,to identify independent risk factors for ACS in CHD patients.Receiver operating characteristic curves were constructed,and area under the curve(AUC)was calculated to evaluate the predictive performance of the independent risk factors and their combinations.Results LAD-FAI,RCA-FAI,and high-sensitivity C-reactive protein(hs-CRP)were independent influencing factors for ACS in CHD patients.The AUC for the prediction of ACS occurrence in CHD patients based on LAD-FAI,RCA-FAI,and elevated hs-CRP values alone were 0.568,0.703,and 0.749,respectively.When these three factors were analyzed in combination,the AUC was 0.815.Conclusion The combined analysis of LAD-FAI,RCA-FAI,and hs-CRP has good predictive performance for assessing the risk of ACS in CHD patients.