Overlapping PCR technology was employed to splice IFN?and HSA genes in vitro. The spliced gene was inserted into Pichia pastoris secretory vector pPIC9K. The IFN?-HSA gene was designed for secretory expression under the control of promoter AOX1 and Mat a signal peptide in pPIC9K. The recombinant plasmid pPIC9K/IFN?-HSA was linearized by restriction enzyme SalI and transformed into Pichia pastoris KM71 by electroporation. The recombinant strains identified by G418 selection and confirmed by PCR analysis were induced by methanol to express fusion protein IFNp-HSA. SDS-PAGE and Western blot analysis of the fusion protein showed that the expressed fusion protein IFNp-HSA with an apparent 90kDa molecular weight had the antigenicity of HSA. The specific activity of culture supernatant was about 640IU/ml assayed by the standard amiviral activity test on WISH cells challenged with VSV virus.

OBJECTIVETo evaluate the curative effect and surigical skills of reconstruction intramedullary nailing in the treatment of ipsilateral femoral neck and shaft fractures.

METHODSFrom January 2007 to January 2013, 13 patients with ipsilateral femoral neck and shaft fractures were treated by reconstruction intramedullary nailing including 11 males and 2 females with an average age of 38.9 yeas old ranging from 25 to 53 years old. For femoral neck fractures,10 cases were basilar neck fracture, 3 were transcervical fractures and according to Garden classification,10 were type II, 2 were type III and 1 was type IV. For feormal shaft fracture, 5 were type I, 4 were type II, 3 were type III and 1 was type IV according to Winquist classification. The location of all 13 feormal shaft fractures were all in the uper 3/4 of the feormal shaft. Complications and postoperative function were recorded and evaluated.

RESULTSEleven patients were followed up for 23.45 months (12 to 36 months). Time from injury to operation was 5.9 days in average. Malunion of femoral neck fracture happened in 1 patient, nonunion of femoral shaft fracture happened in 2 patients. NO femoral head osteonecrosis, infection and malunion of femoral shaft fracture happened. According to Friedman-Wyman classification, 8 cases get good result, 2 get faii and 1 get poor.

CONCLUSIONReconstruction intramedullary nailing is a good choice for the ipsilateral femoral neck and shaft fractures with the advantages of less soft-tissue trauma and complications.

Adult ; Female ; Femoral Fractures ; surgery ; Femoral Neck Fractures ; surgery ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Middle Aged

The selective side-chain cleavage of phytosterol to 4-androstene-3,17-dione(4-AD)and 1,4-androstadiene-3,17-dione(ADD)by Mycobacterium sp.was described.Because of the similarity in chemical structure between 4-AD and ADD,it is difficult to separate them from the fermentation broth.So far,it has been verified that the ADD can be produced by dehydrogenation of 4-AD.In this reaction,3-Ketosteriod-1-Dehydrogenase(ksdD)plays an important role.The gene knocking out method was used to solve the problem.Partial sequence of ksdD was obtained by PCR which was 631bp in length.Then,a targeting vector pUC19-MK was constructed,which was electroporate into the original strain Mycobacterium neoauru.The method of homologous recombination was used to knock out ksdD gene located in the chromosome of Mycobacterium neoauru.In this way,ksdD would lose its enzyme activity.In the result,5 transformants were screened.The experiments of steroid transformation by the transformants were carried out.The productivity of 4-AD reached 17.52% after 144h,which is 192% higher than the original strain.Meanwhile,the productivity of ADD reached 6.12%,which is 89.9% lower than the original strain.

Objective To evaluate the diagnostic value of MRI in brachial plexus injury.Methods Total 98 patients with brachial plexus injury were examined by MRI before operation.Fifty-four of 98 patients MR imaging were obtained by 0.5 Tesla scanner and other 44 patients were obtained by 1.5 Tesla scanner.The scanning sequences include: SE T_1WI,T_2WI,FFE T_2WI and T_2WI SPIR. Exploration of the supraclavicular plexus was carried out and the MR imaging were compared with the operative finding in 63 patients.Thirty-five patients who had not surgery were followed-up.Results MR imaging found pre-ganglionic injuries in 45 patients and post-ganglionic injuries in 56 patients.Pre-and post-ganglionic injuries simultaneously in 16 patients among them.MR imaging can not find injury sings in 13 patients.The positive rate was 86.73%.MR imaging finding of pre-ganglionic injuries include:(1) Spinal cord edema and hemorrhage,2 patients (4.44% ).(2)Displacement of spinal cord,17 patients (37.78%).(3)Traumatic meningoceles,37 patients (82.22%).(4)Absence of roots in spinal canal, 25 patients(55.56% ).(5)Scarring in the spinal cnanl,24 patients (53.33%).(6)Denervation of erector spine,13 patients (28.89%).MR imaging finding of post-ganglionic injuries include:(1)Trunk thickening with hypointensities in T_2WI,23 patients (41.07%).(2)Nerve trunk complete loss of continuity with disappeared of nerve structure,16 patients (28.57%).(3)Continuity of nerve trunk was well with disappearance of nerve structure,14 patients(25.00%).(4)Traumatic neurofibroma,3 patients (5.36%).Conclusion MR imaging can reveal Pre-and post-ganglionic injuries of brachial plexus simultaneously.MR imaging is able to determine the location (pre-or post-ganglionic)and extent of brachial plexus injury,provided important information for treatment method selection.

Objective The aim of this study is to investigate the role of angiotensin-converting enzyme (ACE)gene susceptibility to systemic lupus erythematosus(SLE)by familial studies.Methods PCR-based re- striction fragment length polymorphism(RFLP)was applied to genotype single nucleotide polymorphism(SNP) G261T of the ACE gene.A total of 119 patients with SLE from 119 families were recruited.In addition,316 family members of these patients were also genotyped.A family-based association study was carried out to ex- plore the association between gene polymorphism and SLE.We studied the SNP encoding non-synonymous substitution in the ACE gene with respect to genetic susceptibility to SLE.Results Among 119 SLE patients. the frequency of ACEG261TG,T alleles was 44.8%.55.2% respectively,the frequency of ACEG261T GG,GT and TT genotypes was 13.9%,62.0%,24.1% respectively,Univariate(single-marker)family-based association test(FBAT)demonstrated that variant alleles at the SNP,rs4303,exon 5 of ACE gene were significantly asso- ciated with genetic susceptibility to SLE in Additive Model(Z=2.877,P=0.004),Dominant Model(Z=2.557, P=0.011).Recessive Model(Z=2.202,P=0.028).Transmission-disequilibrium test(TDT)and sib transmission -disequilibrium test(STDT)showed an excess of the allele of T from heterozygous parents to affected offspring or higher frequency of the allele of T in the patients than their normal siblings(X~2=11.66,P=0.001).Conclu- sion Our findings suggest that the ACE gene may he the susceptible gene to SLE in Chinese population,and the individuals carrying ACE-261T allele is significantly associated with susceptibility to SLE.

OBJECTIVETo investigate the etiologic value of diarrheagenic E. coil harboring genomic O island 28(OI-28) containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635), which were related to RTX (Repeat in toxin) toxin family isolated from children with diarrheal disease in Taiyuan.

METHODSIn the study, 257 fecal samples from children with diarrheal disease collected in Shanxi Children's Hospital. Diarrheagenic E. coli and enteropathogenic bacteria were isolated and identified by conventional bacterial culture and typing specific diarrheagenic E. coli (EPEC, EIEC, ETEC and EHEC) diagnostic serum, while diarrheagenic E. coli harboring genomic 01-28 containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635) were detected by PCR and DNA southern blot hybridization.

RESULTS206 strains (80.16%) of enteropathogenic bacteria were detected from 257 children with diarrhea disease, containing 149 strains (57.98%) of diarrheagenic E. coli and 57 strains(22.18%) of other entero-pathogenic bacteria. Among 3 strains (2.01%) of EPEC, 2 strains (1.34%) of ETEC, 2 strains (1.34%) EHEC were detected by typing specific serum, while all of the 142 strains (95.30%) isolated were suspected to be diarrheagenic E. coli. 21 strains (14.09%) of diarrheagenic E. coil harboring genomic O1-28 containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635) were detected by polymerase chain reaction and DNA southen blot hybridization, 8 strains (5.37%) of diarrheagenic E. coli containing only one genomic OI-28 virulence gene, 2 strains (1.34%) of diarrheagenic E. coli containing two genomic OI-28 virulence gene. 21 children with diarrhea diseases caused OI-28-harboring E. coli containing five important putative virulence genes were among 0 to 3 years old (80.95%). These children correlating with OI-28-harboring E. coli did not present special clinical symptoms or signs.

CONCLUSIONThe diarrheagenic E. coil harboring genomic OI-28 was one of the important etiology for children with diarrheal disease in summer season.

Child ; China ; Diarrhea ; microbiology ; Escherichia coli ; genetics ; pathogenicity ; Escherichia coli Infections ; complications ; Genes, Bacterial ; Humans ; Virulence

To examine if polyprotein gene (VP2/VP4/VP3) of Infectious Bursal Disease Virus (IBDV) could be delivered into mammalian cells and expressed using attenuated Salmonella typhimurium as vector. The IBDV polyprotein gene was amplified by RT-PCR and inserted in to pCI, an eukaryotic expression plasmid. The resulting recombinant pCI-VP2/VP4/VP3 was transformed by electroporation into attenuated Salmonella typhimurium strain ZJ111 (dam- and phoP-), which was then use to transfect the Vero cells. Gene specific RT-PCR revealed that VP2/VP4/VP3 was transcribed into mRNA in the Vero cells. Indirect immunofluorscence assay, SDS-PAGE and Western-blot analysis showed that VP2/VP4/VP3 was expressed and the product was immuno-reactive with anti-IBDV serum. This work provides essential precondition for developing a new oral DNA vaccine against IBDV.
Animals ; Cercopithecus aethiops ; Electroporation ; Genetic Vectors ; genetics ; Infectious bursal disease virus ; genetics ; metabolism ; Polyproteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Salmonella typhimurium ; genetics ; metabolism ; Transfection ; Vero Cells ; Viral Proteins ; genetics

BACKGROUNDIn recent years, the number of patients undergoing primary total knee arthroplasty in China has rapidly increased. However, the incidence of primary total knee arthroplasty is unknown. The purpose of this study was to investigate the sex, age and, annual incidence of primary total knee arthroplasty based on 3118 Chinese patients who underwent the procedure during the period of 2000 - 2011.

METHODSTotal knee arthroplasties were performed on 511 males and 2607 females in our hospital during the period of 2000 - 2011. The sex, age, and annual incidence of primary total knee arthroplasty were evaluated.

RESULTSThe annual incidence of primary total knee arthroplasty increased from 35 knees in 2000 to 681 knees in 2011. The average annual percentage increase in incidence was 33.2%. Females accounted for 83.2% of the patients who underwent primary total knee arthroplasty. In both males and females, the highest incidence was observed in the group aged 65 - 74 years.

CONCLUSIONSThis study demonstrated a rapid increase in the incidence of primary total knee arthroplasty in our Chinese study population. The sex and age incidence of primary total knee arthroplasty in our study population differed from those reported in Western populations.

Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; statistics & numerical data ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Osteoarthritis ; surgery ; Sex Distribution ; Young Adult

BACKGROUND: With a clear distinction from traditional computed tomography (CT) imaging with information absorption, phase-contrast CT with synchronous radiation has implemented the microstructure imaging of soft tissues in organisms with an unprecedented imaging mechanism. OBJECTIVE: To explore the synchrotron radiation phase-contrast CT imaging technology in the bone regeneration imaging after bone grafting. METHODS: Four New Zealand white rabbits were used to make a metaphyseal defect model. Then, model rabbits were randomized into a group with calcium phosphate bone grafting and a group with Bio-Oss bone grafting in the defects. The specimens were imaged by the synchrotron radiation phase-contrast CT and stained with hematoxylin-eosin and sirius red 2 weeks after bone grafting. RESULTS AND CONCLUSION: (1) Bio-Oss bone graft material group: Osteoid was observed not only around the graft material but also in the area far from the graft bone material as reticulate structure by the synchrotron radiation phase-contrast CT. Hematoxylin-eosin staining showed a large amount of red osteoid tissues arranged as trabecular bone, and a large amount of osteoblasts with obvious osteogensis. Sirius red-stained pathological sections were largely stained yellow, and there were round or oval osteoblasts with strongly expressed type I collagen. (2) Calcium phosphate bone graft material group: There was no reticulate structure shown by the synchrotron radiation phase-contrast CT, and the creep of osteoid tissues was only around the bone graft. Hematoxylin-eosin staining showed a large amount of red osteoid tissues, and sirius red-stained pathological sections were stained yellow and red. To conclude, the synchronous radiation phase-contrast CT can clearly display the regenerated structure of bone grafts.

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling ; Panax ; genetics ; Saponins ; biosynthesis ; Sequence Analysis, RNA