Copy number variations (CNVs) refer as a DNA segment that is 1 kb or larger and is presented at a variable copy number in comparison with a reference genome. Classes of CNVs include insertions, deletions, duplications and their complex combinations. Because they widely distributed in the genome with some important characteristics, such as heritable, relative stable and heterogeneity, CNVs are considered as novel genomic polymorphism markers. And the alteration of gene dosage which resulted from CNVs could change phenotype, so a novel CNV genome-wide association analysis (CNV-GWAS) strategy appeared recently and began to used for identifying susceptible genes of complex diseases. It was approved that it could complement the tranditional genome-wide association studies based on single nucleotide polymorphisms. Therefore, genomic structure variances are favorable for revealing the molecular mechanisms and genetic foundation of complex diseases.

Objective To investigate the relationship between osteoprotegerin ( OPG ) gene polymorphisms and ischemic stroke etiological subtypes, as well as the extent and distribution of cerebral atherosclerosis ( AS) lesions.Methods Patients with ischemic stroke included 285 cases of large-artery atherosclerosis (LAA), 91 cases of small-artery occlusion (SAO) and 42 cases of purely AS, and 165 healthy controls were enrolled in this study.The LAA group was respectively divided into 3 subgroups according to the number and the distribution of stenostic vessels.Genotyping of three single nucleotide polymorphisms (SNPs;rs2073617, rs3134069, and rs3102735) in the promoter region of the OPG gene was performed by polymerase chain reaction-restriction fragment length polymorphism.Results Regarding the three SNPs of OPG gene, the frequence of genotype CC/CT and the prevalence of allele C of rs3102735 were higher in the LAA group contrasting with the control group ( 24.04% vs 14.85%, 44.21% vs 27.88%,χ2 =10.758, 11.804, P =0.001,0.024).However, comparisons of other frequences of genotypes or alleles did not reveal any significant differences among the LAA group, the SAO group, the AS group and the control group, as well as among different subgroups of LAA group.Haplotype analysis revealed that the frequencies of haplotype C-C-T in LAA group and SAO group were significantly lower ( 0.023, 0.017 vs 0.068,χ2 =10.399, 5.841,P=0.001, 0.016), while that of haplotype T-A-C was significantly higher in SAO group(0.043 vs 0.016,χ2 =4.708, P=0.030) compared with controls.Conclusions Our findings indicate that OPG gene polymorphisms might be associated with increased susceptibility to LAA ischemic stroke.But we fail to show association of OPG gene with the extent and distribution of AS.

Zinc-finger proteins have been widely studied due to their highly conserved structures and DNA-binding specificity of zinc-finger domains. However, researches on the zinc-finger proteins from microorganisms, especially those from prokaryotes, are still very limited. This review focuses on the latest progress on microbial zinc-finger proteins, especially those from prokaryotes and the application of artificial zinc-finger proteins in the breeding of robust strains. Artificial zinc-finger proteins with transcriptional activation or repression domain can regulate the global gene transcription of microbial cells to acquire improved phenotypes, such as stress tolerance to heat, ethanol, butanol, and osmotic pressure. Using the zinc-finger domain as DNA scaffold in the construction of enzymatic system can enhance the catalytic efficiency and subsequently the production of specific metabolites. Currently, zinc-finger domains used in the construction of artificial transcription factor are usually isolated from mammalian cells. In the near future, novel transcription factors can be designed for strain development based on the natural zinc-finger domains from different microbes, which may be used to regulate the global gene expression of microbial cells more efficiently.
Bacteria ; metabolism ; DNA ; chemistry ; Protein Engineering ; Transcription Factors ; chemistry ; Transcriptional Activation ; Zinc Fingers

Objective To construct a eukaryotic expression vector in Pichia pastoris containing human tissue factor( hTF) gene,in order to achieve high level secretory expression in extracellular.Methods Expression plasmid, pGAPZaA-hTF, was constructed by inserting the synthesized sequence encoding human extracellular tissue factor into yeast expression vector pGAPZaA and transformed into Pichia pastoris SMD1168H with electroporation.Having been selected by Zeocin, transformants containing hTF cDNA were expressed in YPD, extracellular proteins were detected by SDS-PAGE and confirmed by Western bolt.Results Successfully constructed the recombinant pGAPZaA-hTF expression system in Pichia pastoris.SDS-PAGE showed that the molecular weight of the expression product was about 37 -40 kDa.Western-blot indicated that it was human extracellular tissue factor.The crude yield of total protein in medium was up to 1 g/L, more than 80% of which was hTF.Conclusion Truncated rTF gene is expressed in Pichia pastoris and the active products are secreted into the medium which have the same activation as the native TF.

BACKGROUND:Recent reports only concern vertebroplasty for mild to moderate vertebral compression fractures, but seldom address vertebroplasty for severe vertebral compression fractures such as vertebral colapse. OBJECTIVE:To evaluate the efficacy and imaging features of percutaneous vertebroplasty for severe vertebral compression fractures. METHODS:A total of 25 patients underwent single-level vertebroplasty for vertebral compression fractures. Imaging features were then analyzed including location, pattern of compression, extent of colapse, pre- and post kyphotic angle and adjacent disc height before and after vertebroplasty. RESULTS AND CONCLUSION:60% (16/25) patients involved the thoracolumbar junction. The height of colapsed vertebral body was 14%-30% of original vertebral body. The average height of colapse was 5.17 mm or 22% of original vertebral body. Kyphotic angle before vertebroplasty ranged from 0-33° (averagely 16°) with an average correction of 12° after vertebroplasty. Average disc height before vertebroplasty was 7.3 mm above and 7.7 mm below. Visual analogue scale score was significantly higher preoperatively than that postoperatively, showing significant difference in pain improvement (P< 0.015). These data suggest that percutaneous vertebroplasty is safe and effective in the treatment of single level severe vertebral compression fractures.

Objective To collect the basic information on publications in Chinese Journal of Perinatal Medicine,and understand the current situation and set the goals for the future. Methods Information on all publications in this journal from 1998 to 2007 was collected.The distribution of all publications,Price Index and relative data were analyzed. Results There were 1368 publications altogether during the past 10 years.The first six kinds of papers were original articles(508,31.7%),short articles(258,18.9%),case reports(194,14.2%),reviews(122,8.9%),experimental studies(90,6.6%)and brief communications(83,14.2%).47.9% of all publications were from Beijing,Guangdong and Shanghai.The first three areas who had the most submissions were Beijing,Shandong and Guangdong in 2007.The citation frequency analysis showed that 708 publications were cited at 1east once and only two were cited over 80 times.53.4% of all cited publications were original articles and experimental studies. Conclusions Original articles and short articles are the two main types of publications in this journal,however original articles and experimental studies contributed to half of the publications being cited.Publications in this journal are focused in few areas and this might be related to the number of submissions.

Objective:To observe the clinical efficacy of warm needling moxibustion plus tuina in treating knee osteoarthritis (KOA) due to cold-dampness blocking collaterals.
Methods:Forty-eight KOA patients were randomized into 2 groups by their visiting sequence, 24 cases in each group. The treatment group was intervened by warm needling moxibustion plus tuina, while the control group was treated with acupuncture plus tuina. The two groups were both treated once a day, 30 min for each session, 10 d as a treatment course, totally for 3 treatment courses. The visual analogue scale (VAS) and Lysholm knee scoring scale (LKSS) were observed before and after the treatment to evaluate the clinical efficacy.
Results:After 3 treatment courses, the VAS and LKSS scores were significantly changed in the two groups (P<0.01), and the between-group differences were also statistically significant (P<0.01). The markedly-effective rate was 83.3% in the treatment group versus 66.7% in the control group, and the difference was statistically significant (P<0.05).
Conclusion:Warm needling moxibustion plus tuina can produce a more significant efficacy than acupuncture plus tuina in treating KOA due to cold-dampness blocking collaterals.

AIM: To demonstrate the mechanisms underlying cardioprotection induced by ischemic postconditioning(I-postC) via studying the alteration of calreticulin(CRT)/calcineurin(CaN) signaling pathway in rat heart subjected to ischemia/reperfusion(I/R).METHODS: The model of myocardial I/R injury in vivo was made by occluding the left anterior descending artery for 45 min followed by 24 h of reperfusion in Wistar rats.Hemodynamics and activity of lactate dehydrogenase(LDH) and creatine kinase-MB(CK-MB) in plasma were measured.Myocardial infarct size was measured by 2,3,5-triphenyltetrazolium chloride(TTC) staining and cardiomyocyte apoptosis was detected using in situ TDT-mediated dUTP nick end labeling(TUNEL).The activity of CaN,the expressions of CaN and CRT in myocardium were detected by enzyme reaction phosphorus measurement and Western blotting analysis,respectively.RESULTS: Cyclosporin A,the inhibitor of CaN,limited significantly myocardial infarct size and cardiomyocyte apoptosis induced by I/R,but had no significant effect on cardiac function.I-postC ameliorated significantly the cardiac dysfunction induced by I/R.Compared with those in I/R group,the myocardial infarct size,the LDH and CK-MB activities in plasma and the cardiomyocyte apoptotic index were significantly reduced in I-postC group.In addition,I/R-induced upregulation of CaN activity,CaN and CRT expression were relieved by I-postC.No significant difference was found between I-postC and ischemic preconditioning groups.I-postC had stronger protective effect on the reperfused heart compared with cyclosporin A.CONCLUSION: The findings indicate that I-postC protects myocardium against I/R injury,at least in part,via inhibiting the CRT/CaN signaling pathway.

Objective To understand the epidemic trend and drug susceptibility of pathogenic bacteria in children with lower re-spiratory tract infection to provide the basis for clinical rational drug use.Methods The sputum culture specimens in these two hospitals during 2013 were performed the pathogenic bacterial culture and the drug susceptibility test respectively.The results were conducted the comparative analysis.Results 6124 strains of bacteria were isolated from the children lower respiratory tract sputum specimens in our hospital during 2013,including 5 121 strains of Gram negative bacteria,accounting for 62.7%,which were domina-ted by Haemophilus influenzae,Klebsiella pneumoniae,Escherichia coli,Bauman Acinetobacter and Pseudomonas aeruginosa;Gram positive bacteria were 2734 strains,accounting for 33.5%,the top two were Streptococcus pneumoniae and Staphylococcus aureus;311 strains of fungi,accounting for 3.8%,and Candida albicans was predominant.1600 strains of bacteria were isolated from the lower respiratory tract sputum specimens in Nanjing Brain Hospital during 2013,including 1 134 strains of Gram negative bacteria, accounting for 70.9%,which were dominated by Klebsiella pneumoniae,Escherichia coli,Pseudomonas aeruginosa and Acinetobact-er Bauman;296 strains of Gram positive bacteria,accounting for 18.5%,Staphylococcus aureus was predominant;170 strains of fungi,accounting for 10.6%,Candida albicans was predominant.The sensitive drugs to Gram-negative bacilli were piperacillin /tazobactam and imipenem,while the sensitive drugs to Gram-positive bacilli were linezolid and vancomycin.Conclusion The Gram negative bacteria are the main pathogens of lower respiratory infection in children,and strengthening bacterial culture and drug re-sistance monitoring is necessary for rational use of antibacterial drugs.

Background Mesenchymal stem cells (MSCs) have been applied in basic and clinical researches of organ transplantation.Our previous study showed that intravenous injection of MSCs prolonged corneal allograft survival in rat.However,the effect of local administration of MSCs on corneal allograft rejection remains unclear.Objective The aim of this study was to investigate the effect of subconjunctival injection of MSCs on corneal allograft rejection in rat model of keratoplasty.Methods MSCs were isolated and cultured from femur and tibia bone marrow of clean Wistar rats,and then the cells were identified by induced differentiation of osteoblast and adipocyte.The third generation of MSCs was used in subsequent experiment.Allogenic penetrating keratoplasty was performed with the bilateral corneas of 20 Wistar rats as donor grafts and the right eyes of 40 Lewis rats as recipients.PBS 0.1 ml containing 2 × 106 MSCs and 0.1 ml PBS only was subconjunctivally injected immediately and postoperative 3 days respectively in randomized two groups,and another 6 normal Lewis rats served as the normal control group.Corneal rejection response was evaluated under the slit lamp after surgery based corneal opacity,edema and neovascularization,and the grafts were scored according to the criteria of Larkin.The corneal samples were extracted from 12 rats of the PBS control group and the MSCs group separately 10 days after surgery.The relative expressions of Th1 cytokines (interferon-γ [IFN-γ] mRNA and interleukin-2 [IL-2] mRNA) and Th2 cytokines (IL-4 mRNA and IL-10 mRNA) were detected by real-time quantitative PCR.Protein levels of IL-4 and IL-10 proteins in the corneas were assayed by ELISA.All experimental protocols involving rats were approved by the laboratory animal care and use committee of the Tianjin Medical University and treated with the ARVO statement for the use of animals in ophthalmic and vision research.Results The cells grew well with the orange stain for alizarin red in differentiated the osteoblasts and red stain for Oil red O in differentiated adipocytes.The survival time of corneal graft in the MSCs group was (11.8±1.6) days,it was significantly longer than (9.6±1.4) days in the PBS control group (P=0.004).The levels of IL-4 mRNA and IL-10 mRNA in the MSCs group were significantly higher than those in the PBS control group (both at P =0.00);while the levels of IFN-γ mRNA and IL-2 mRNA were not significantly different between the groups (both at P＞0.05).The IL-10 protein contents were (22.74 ±7.06),(68.40±12.83) and (215.41 ±44.66)pg/ml in the normal control group,PBS control group and MSCs group,showing significant difference among the three groups (F =55.06,P =0.00) and a significant increase in the MSCs group compared with the PBS control group and the normal control group (both at P ＜ 0.05).Conclusions Subconjunctival injection of MSCs prolongs the survival time of cornea allograft in penetrating keratoplasty probably by modulating the balance between Th1 and Th2 cytokines,especially by up-regulating Th2 cytokines.