Objective To observe the therapeutic efficacy and action mechanism of muscle-region alignment electroacupuncture in treating post-stroke shoulder pain. Method Eighty patients were randomized into a muscle-region alignment needling group and a conventional acupuncture group. The Short-form McGill Pain Questionnaire (SF-MPQ), and serum levels of IL-6, TNF-?, and NO were majorly observed before and after the treatment. Result The muscle-region alignment electroacupuncture and conventional acupuncture both obviously reduced the SF-MPQ score and down-regulated the serum levels of IL-6, TNF-?, and NO, and the decreases by the muscle-region alignment electroacupuncture were more significant than that by the conventional acupuncture. Conclusion The action of muscle-region alignment electroacupuncture in treating post-stroke shoulder pain is plausibly by down-regulating serum levels of IL-6, TNF-?, and NO, reducing or inhibiting the production of inflammatory factors and restraining inflammation.

Objective To analyze the genotype and plasmid transfer of Enterobacteriaceae carring blaNDM‐1 with blaIMP‐4 or blaKPC‐2 .Methods From April 2012 to October 2014 ,a total of 33 non‐repeatitive carbapenem‐resistant Enterobacteriaceae ( including Imipenem‐resistant , meropenem‐resistant or Ertapenem‐resistant) were isolated from 5 hospitals in Wenzhou and Hangzhou . Identification and antimicrobial susceptibility test were performed using Vitek 2 Compact automatic microbiology analyzer . Phenotypes of carbapenemase were screened using modified Hodge test and ethylenediamine tetraacetic acid‐disk synergy test .Extended spectrum βlactamase test was determined by the double disk combination test which was recommended by Clinical and Laboratory Standards Institute .AmpC activity was tested by a three‐dimensional Cefoxitin method .Drug resistant genes including blaNDM‐1 and linkage of ISAba125‐NDM were detected by polymerase chain reaction (PCR) .The purified PCR products were cloned and sequenced .Plasmid conjugation experiment and elimination method were carried out to test partial bacterial strain and K . pneumoniae carrying blaNDM‐1 with blaIMP‐4 or blaKPC‐2 .Results Of the 33 non‐repeatitive carbapenem‐resistant Enterobacteriaceae ,28 were strains of K .pneumoniae ,1 strain of K . oxytoca,2strainsof Escherichiacoli,1strainof K.planticolaand1strainof E.cloacae.Thirteenstrains were isolated from Hospital of Sir Run Run Shaw of Zhejiang University ,thirteen from Wenzhou Hospital of Traditional Chinese Medicine ,one from Wenzhou People′s Hospital ,three from the First Affiliated Hospital of Wenzhou Medical University and three from Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine .Thirty‐one strains were confirmed as carbapenemase‐producing with 24 of blaKPC‐2 ,3 of blaNDM‐1 ,1 of blaNDM‐5 and 3 of blaIMP‐4 .Among them ,one strain carried blaNDM‐1 with blaIMP‐4 and one strain carried blaNDM‐1with blaKPC‐2 ,respectively .The plasmid transfer and conjugation experiment was performed between strains carrying blaNDM‐1 and Escherichia coli EC600 or K . pneumoniae ATCC13833 and genes of blaNDM‐1 and ISAba125‐NDM were obtained .Conclusions blaKPC‐2 gene is the popular carbapenemase genotype .blaNDM‐1 or blaNDM‐5 may be correlated with linkage gene of ISAba125‐N DM .Coexistence of blaNDM‐1 carrying blaIMP‐4 or blaKPC‐2 is detected in the same strain , respectively . Enough importance should be attached to the strains ,because most of them are multiple drug resistance with related genes located in the plasmid which is easily spread between strains .

Objective:To explore the molecular pathogenesis of a family with hereditary factor Ⅴ (FⅤ) deficiency.Methods:All the exons, flanking sequences, 5′ and 3′ untranslated regions of the F5 of the proband, and the corresponding mutation sites of the family members were analyzed via direct DNA sequencing. The CAT measurement was used to detect the amount of thrombin produced. The ClustalX software was used to analyze the conservation of mutation sites. The online bioinformatics software, Mutation Taster, PolyPhen-2, PROVEAN, LRT, and SIFT were applied to predict the effects of mutation sites on protein function. The Swiss-PdbViewer software was used to analyze the changes in the protein model and intermolecular force before and after amino acid variation.Results:The proband had a heterozygous missense mutation c.1258G>T (p.Gly392Cys) in exon 8 of the F5, and a heterozygous deletion mutation c.4797delG (p.Glu1572Lys fsX19) in exon 14, which results in a frameshift and produces a truncated protein. Her grandfather and father had p.Gly392Cys heterozygous variation, whereas her maternal grandmother, mother, little aunt, and cousin all had p.Glu1572LysfsX19 heterozygous variation. The ratio of proband's thrombin generation delay to peak time was significantly increased. Conservation analysis results showed that p.Gly392 was located in a conserved region among the 10 homologous species. Five online bioinformatics software predicted that p.Gly392Cys was pathogenic, and Mutation Taster also predicted p.Glu1572Lys fsX19 as a pathogenic variant. Protein model analysis showed that the replacement of Gly392 by Cys392 can lead to the extension of the original hydrogen bond and the formation of a new steric hindrance, which affected the stability of the protein structure.Conclusion:The c.1258G>T heterozygous missense mutation in exon 8 and the c.4797delG heterozygous deletion mutation in exon 14 of the F5 may be responsible for the decrease of FⅤ levels in this family.

Objective:To investigate the distribution of single nucleotide polymorphism and haplotype of human leucocyte antigen G 3′untranslated region gene, which possibly could be predictive roles in unexplained recurrent spontaneous abortion patients.Methods:Case-control method was used in this study. 70 cases of pregnant women with unexplained recurrent spontaneous abortion and 54 cases of prenatal examination women whose peripheral blood and serum were collected in Wenzhou Hospital of Chinese Traditional Medicine were recorded from June 2017 to July 2018. Blood gene DNA was extracted by centrifuge column and was amplified by polymerase chain reaction. Sanger sequencing method was used for genotyping. The genotypes frequency, linkage imbalance analysis and haplotypes construction of SNPs were analyzed by SHEsis online software and Phase software. Serum soluble HLA-G concentration was detected by ELISA.Results:There were eight SNPs, including 14bp ins/del,+3003C/T,+3010G/C,+3027A/C,+3035C/T,+3142C/G,+3187A/G and+3196C/G, were detected in both the URSA group and the control group. Results showed that the distribution differences of+3010G/C,+3142C/G and+3187A/G between the two groups were statistically significant (χ 2=8.514, P=0.004; χ 2=0.552, P=0.021; χ 2=8.183, P=0.005) .The C allele at the+3010G/C site and the G allele at the +3142C/G site might be risk factors for URSA ( OR=2.131, 95 %CI=1.278-3.552, χ 2=8.514, P=0.004; OR=1.813, 95 %CI=1.091-3.013, χ 2=0.552, P=0.021) ;the G allele at +3187A/G site might be a protective factor for URSA ( OR=0.476, 95 %CI=0.285-0.794, χ 2=8.183, P=0.005) .Haplotype analysis revealed that UTR-1 (DTGCCCGC) might be a protective factor for URSA ( OR=0.497, 95 %CI=0.295-0.837,χ 2=6.987, P=0.008), while UTR-3 (DTCCCGAC) might be a risk factor for URSA ( OR=1.732, 95 %CI=1.009-2.974, χ 2=3.998, P=0.045).The frequency of UTR-1/UTR-1 homozygous in URSA patients was lower than that in normal patients obviously( OR=0.381, 95 %CI=0.165-0.879, χ 2=5.292, P=0.024), which might be a protective factor for pregnancy. No association was found between serum soluble HLA-G and HLA-G 3′UTR gene haplotypes in URSA ( t=1.578, P=0.119) . Conclusions:HLA-G 3′UTR gene polymorphism and haplotypes are correlated with URSA. The study lays a foundation for future research and provides a basis for clinical individualized medicine.