Objective To review the surgical managements of patients with traumatic false aneu- rysms in the extremities.Methods From January 1990 to April 2006,17 patients with traumatic false aneurysms in the extremities were admitted into our hospital.Fourteen patients were treated by vascular repair including vascular repair in seven cases,end to end anastomosis in one,synthetic grafting in one, autogenous vein grafting in one,and direct ligation in four.Three patients were treated nonoperatively, but with local compressive dressing.Results There were no deaths or gangrenes in all cases.The clinical manifestations vanished after the treatment.The mean follow-up period was 13.2 months.The function of the injured extremities recovered satisfactorily.Conclusion Different types of traumatic false aneurysms should be managed by different therapeutic procedures after the diagnoses is made.

AIM:To investigate the characteristic of T-cell acute lymphocytic leukemia 1 (TAL1) gene expres-sion in acute myeloid leukemia (AML) cell lines and in primary AML cells from de novo AML patients with different sub-types. METHODS:Real-time PCR was used to determine the expression of TAL1 mRNA in acute leukemia cell lines (Jurkat, CCRF-CEM, HL-60 and NB4 cell lines) and peripheral blood mononuclear cells from 47 newly diagnosed AML patients. Twelve healthy individuals were served as healthy control group. RESULTS:A significantly increased level in TAL1 mRNA was found in AML cell lines (HL-60 and NB4), T-cell acute lymphacytic leukemia (T-ALL) cell lines (Jur-kat, CCRF-CEM) and primary AML cells compared with the healthy controls. Over-expression of TAL1 was found in all detected AML subtypes, the highest level of TAL-1 mRNA was found in AML-M1 and AML-M5 subtype ( P <0.05). CONCLUSION:High expression of TAL1 in AML might influence the differentiation and proliferation of myeloid cells, further investigation needs to confirm whether it would be as a biomarker for pathogenesis of AML.

Objective To verify the performance characteristics of the human SLCO1B1 & APOE gene detection kit (PCR-fluorescent probe technique).Methods The verification protocol for the human SLCO1B1 & APOE gene detection kit manufactured by Wuhan YZY Medical Science and Technology Co.,Ltd.was designed by referring to the requirements of professional standard and pertinent literatures,and its accuracy,specificity,lowest detection limit and anti-interference ability were compared with those of sequencing technique (gold standard).Results The results coincidence of 20 clinical samples from fluorescence PCR and sequencing technique was 100％,which met the requirement of accuracy.Forty clinical samples with wild type loci determined by fluorescence PCR were verified by sequencing and no any mutation was detected,which met the requirement of specificity.The lowest detection limit of the kit was 10 ng/μL.Three interfering substances,including hemoglobin,bilirubin and triglyceride,were individually added into the blood samples with known genotype,and the results determined by the kit were coincident with that without interfering substances,which met the requirement of anti-interference ability.Conclusion The verification parameters of the human SLCO1B1 & APOE gene detection kit are basically consistent with the manufacturer's statement,indicating that the kit meets the requirements of the relevant quality management and may be applied to the detection of clinical samples.

Objective To test the changes of the potassium(K+)and magnesium(Mg2+)concentrations in vitreous humor of rabbits along with postmortem interval(PMI)under different temperatures, and explore the feasibility of PMI estimation using mixed-effect model. Methods After sacrifice, rabbit carcasses were preserved at 5℃, 15℃, 25℃ and 35℃, and 80-100μL of vitreous humor was collected by the double-eye alternating micro-sampling method at every 12 h. The concentrations of K+and Mg2+in vitreous humor were measured by a biochemical-immune analyser. The mixed-effect model was used to perform analy-sis and fitting, and established the equations for PMI estimation. The data detected from the samples that were stoned at 10℃, 20℃ and 30℃ with 20, 40 and 65 h were used to validate the equations of PMI estimation. Results The concentrations of K+and Mg2+[f(x,y)] in vitreous humor of rabbits under different temperature increased along with PMI(x). The relative equations of K+and Mg2+concentration with PMI and temperature under 5℃~35℃ were fK+(x,y)=3.413 0+0.309 2 x+0.337 6 y+0.010 83 xy-0.002 47 x2 (P<0.000 1), and fMg2+(x,y)=0.745 6+0.006 432 x+0.033 8 y(P<0.000 1), respectively. It was proved that the time of deviation for PMI estimation by K+and Mg2+was in 10 h when PMI was between 0 to 40 h, and the time of deviation was in 21 h when PMI was between 40 to 65 h. Conclusion In the ambient temperature range of 5℃-35℃, the mixed-effect model based on temperature and vitreous humor sub-stance concentrations can provide a new method for the practical application of vitreous humor chemi-cals for PMI estimation.

OBJECTIVETo elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein(M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012.

METHODSA total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos,myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT-PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains.

RESULTSA total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained from 30 positive samples and were divided into 3 distinct clades or sub-clades(1.2.1, 1.2.2 and 2) by phylogenetic analysis, 5, 7 and 2, respectively. The M2 genes and Hemagglutinin(HA) genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains.

CONCLUSIONThe M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2.2 had become dominant epidemic strain in this region.

Animals ; Birds ; virology ; Chickens ; virology ; China ; Evolution, Molecular ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; Influenza in Birds ; virology ; Phylogeny ; Poultry ; virology ; Viral Matrix Proteins ; genetics

Objective:To investigate the expression of immunoglobulin A (IgA) in human mesangial cells (HMCs).Methods:The HMCs were cultured.The subcellular location of IgA was detected by immunofluorescence staining;the transcripts of Ig α,Ig κ and Ig λ constant region were detected by reverse transcription-polymerase chain reaction (RT-PCR) and further analyzed by DNA sequencing.The expressions of Igαt and Ig λ were detected at transcription level by Western blot after the cytoplasmic protein extraction.The culture supernatant was collected to explore whether IgA could be secreted out of the cell and the protein was further analyzed by mass spectrometry after being purified by affinity chromatography with jacalin-sepharose.The results of DNA sequencing and mass spectrometry were aligned with the mRNA and amino acid sequences in the National Center of Biotechnology Information (NCBI) data-base.Results:By immunofluorescence staining,we detected the presence of IgA heavy chain Ig α,light chain,both Ig κ and Ig λ in expressions of transcripts of Ig α1,Ig α2,Ig κ and Ig λ in the HMCs and the alignment of the sequences of the RT-PCR products with those of the Ig Cα1,Ig Cα2,Ig κ and Ig λ mRNA in the NCBI database exhibited that the similarities were 99％,97％,98％ and 97％,respectively.Western blot showed Ig α and Ig λ expressions in the cell lysate and secretion of Ig α1 and Ig α2 heavy chains in cell culture supernatant.To further explore the protein that secreted into the supernatant,after supernatant affinity chromatography with jacalin-sepharose,the proteins were separated by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) and the band approximating to 65 000 was cut and sent to mass spectrometry.The results were aligned with the amino acid sequences of Ig α1 and Ig α2 constant region in NCBI database,showing that amino acids between No.52 and No.104,amino acids between No.154 and No.221,amino acids between No.276 and No.327 from Ig Cα1 and amino acids between No.52 and No.113,amino acids between No.151 and No.204,amino acids between No.251 and No.314 from Ig Cα2 were the same with those derived from B cells.Conclusion:Our findings suggested that HMCs could synthesize and secret IgA.

Objective To study the effect of panatinib on cholangiocarcinoma(CCA)and its molecular mechanism.Methods QBC939 cells were divided into control group(no treatment),low dose group(0.1 μmol·L-1 panatinib treatment),medium dose group(0.5μmol·L-1panatinib treatment),high dose group(1.0 μmol·L-1 panatinib treatment)and high dose+microRNA-92b-3 group(after treatment with 1.0 μmol·L-1 panatinib transfected miR-92b-3p mimic),high-dose+miR-92b-3p mimic+overexpressed homologous domain interacting protein kinase 3 plasmid(oe-HIPK3)group(after treatment with 1.0 μmol·L-1 panatinib,miR-92b-3p mimic and oe-HIPK3),mimic-NC group(transfected miR-92b-3p NC),and miR-92b-3p mimic group(transfected miR-92b-3p mimic)were transfected.Cell proliferation was detected by cell counting kit 8(CCK-8);cell migration was detected by Transwell,the relative expression level of protein was detected by Western blot.Results Cell proliferation rates of control group,high-dose group,high-dose+miR-92b-3p mimic group,and high-dose+miR-92b-3p mimic group mimic+oe-HIPK3 group were(76.58±8.56)％,(61.85±7.77)％,(74.54±7.68)％and(58.63±6.87)％,respectively;the number of cells migrated were 185.32±20.14,132.33±18.99,168.23±19.85 and 138.66±15.95,respectively;phosphorylated phosphatidyl inositide 3-kinase(p-PI3K)/PI3K ratios were 1.00±0.23,0.68±0.09,0.91±0.18 and 0.60±0.08,respectively;phosphorylated protein kinase B(p-AKT)/AKT ratios were 1.00±0.25,0.61±0.08,1.12±0.28 and 0.72±0.14,respectively.The above indexes were compared with those of the control group in the high-dose group,and those of the high-dose+miR-92b-3p mimic group in the high-dose+miR-92b-3p mimic group in the oe-HIPK3 group.There were statistically significant differences(all P＜0.05).Conclusion Panatinib can effectively inhibit the evil biological behavior of cholangiocarcinoma,which may be related to inhibiting the level of miR-92b-3p and promoting HIPK3-mediated PI3K/AKT signaling pathway.

The purpose of this study was to investigate the regulatory effects of biofilm associated non-coding small RNA(sRNA)00085 on the survival and environmental fitness of Listeria monocytogenes.Homologous recombination technology was used to construct a deletion mutant strain(△sRNA 00085)and a complementary strain(C △sRNA 00085)of the sRNA00085 target gene.The differences in biological characteristics were compared among the standard strain,△sRNA 00085,and C△sRNA 00085.The deletion of sRNA00085 led to a significant decrease in biofilm formation capacity and sensitivity to several antibiotics,including penicillin,piperacillin,doxycycline,tetracycline,vancomycin,and cotrimoxazole.However,only the minimum inhibitory concentration(MIC)of tetracycline exhibited a significant decrease in △sRNA00085.Meanwhile,the decreased biofilm formation and antibiotic resistance of the sRNA00085 mutant were restored in the C△sRNA00085 strain.Furthermore,we investigated the transcription levels of tetracycline resistance-related genes in L.monocytogenes.Down-regu-lated transcription of the tetS gene but no significant difference in transcription of the tetA gene were observed in △sRNA 00085 compared with the standard strain and C△sRNA00085.Moreover,the elimination of sRNA00085 did not affect bacterial growth ability or sensitivity to disinfectants.These findings highlight that sRNA00085 plays an important role in the environ-mental adaptability of L.monocytogenes by affecting bacterial biofilm formation and resistance.

Objective To elucidate the genetic diversifications of avian influenza subtype H5N1 viruses in the boundary regions of Yunnan province during 2009 to July,2011.Methods Swab samples were collected from foreign poultry and wild birds in boundary regions of Yunnan province during 2009 to July,2011 and tested by H5/N1 subtype-specific multiplex RT-PCR.The HA genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD 18-T vectors for sequencing.Both alignment and phylogenetic analysis were performed with sequences of the known reference strains.Results Fifteen different HA sequences were obtained from 36 representative positive samples and could be divided into 2 distinct Clades (2.3.2 and 2.3.4).Through phylogenetic analysis,Clade 2.3.2 and 2.3.4 could then be further divided into 3 ( Ⅱ -1 to Ⅱ -3 ) and 2 smaller clades ( Ⅰ -1 and Ⅰ -2),respectively.The viruses of Clade 2.3.2 Ⅱ -1 and Ⅱ-2 were new variant strains of H5N 1 virus.The cleavage sites of HA from positive samples all possessed molecular characterization of highly pathogenic avian influenza virus.Mutation of key amino acids had been found among receptor binding sites,potential glycosylation sites,neutralizing epitopes and others.Conclusion It seemed evident that the H5N1 subtype viruses showed genetic diversifications and had undergone the evolution progress of multi-clade (2.3.2,2.3.4) to single caide (2.3.2) in the boundary regions of Yunnan province,during 2009 to July,2011.