DC-12 is a Bacillus strain with strong fibrinolytic activity was screened from Douchi.A novel breeding technology——whole genome shuffling was applied to enhance the production of fibrinolytic enzyme.First of all,a candidate mutant library was constructed by treating DC-12 with ultraviolet rays andhNO2,respectively.Based on studying the optimum conditions for protoplast preparation and regeneration of DC-12,multi-parental whole genome shuffling with 4 strains was conceded by protoplasts electrical fusion,then combines with the screening method of double-inactive protoplast,two strains withhigher yield of fibrinolytic enzyme were obtained effectively,which can descend stably after five generations,and their enzyme production reached 2710 IU/ml,which was increased by 4~5 fold compared with the mutants,respectively.

0.05).It was revealed that all the patients followed up 3 to 6 months after treatment were free from complaints of the involved teeth,with nomal food intake and mastication.No abnormal adjacent tissues of dental roots were detected by imaging examination.Conclusion For the vital teeth,the similar clinical effect may be obtained by one-visit RCT and multi-visit RCT.

Based on the principle of evidence-based medicine, the efficacy and safety of consciousness-restoring and orifice-opening (CROO) acupuncture manipulation in treating stroke are systematically assessed. All reports of clinical trials of CROO acupuncture manipulation in treating stroke were collected and random-effect model of Meta-analysis was performed by using the statistical software of Review Manager (RevMan4.1). Sensitivity and heterogeneity were analyzed and tested; funnel-plot analysis was used to identify the publication bias. Comprehensive efficacy (Odds Ratio, OR) of CROO acupuncture manipulation in treating stroke was: OR of cure and markedly effective rate between groups being 3.65 and 95% confidence interval (CI) in the range of 1.70～7.83, OR for decreasing long-term death rate being 0.21 and 95% CI in the range of 0.08～0.52. Stratified analysis showed that OR of stroke sequela was 4.01 and OR of cerebral vascular infarction was 4.45. Adverse drug reaction was not found in all of the reports. Funnel-plot analysis showed the possible existence of publication bias. [ Conclusion]CROO acupuncture manipulation has certain effect in treating stroke, especially in treating acute stage of cerebral vascular infarction, and could reduce long-term death rate of stroke. Heterogeneity may result from the difference of trial designs and inclusive criteria. Funnel-plot analysis shows the possible existence of publication bias. This conclusion is rather limited and will get more evidence if multi-center, double-blinded, randomized and controlled trials could be carried out.

Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells. The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells, and the relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells. By using magnetic active cell sorting (MACS), we isolated a population of CD44(+)/CD133(+) prostate cancer cells that display stem cell characteristics from PC3 cell line. Immunohistochemistry revealed positive expressions of CD44, CD133 and α(2)β(1)-integin in the isolated cells. CCK-8 analysis showed that isolated cells had a strong proliferative ability. The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing, indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line. Western blotting exhibited that the isolated cells had higher experession of Nanog, an embryonic stem marker, as compared with PC3 cells. Our study showed that Nanog might be helpful in sustaining the self-renewal and the undifferentiation of prostate cancer stem cells, and may serve as a marker for prostate cancer stem cells for isolation and identification.

Animals ; Dose-Response Relationship, Drug ; Perchlorates ; toxicity ; Quaternary Ammonium Compounds ; toxicity ; Rabbits

The folate metabolic pathway plays important roles in cellular physiology by participating in nucleotide synthesis, DNA repair and methylation, and maintenance and stability of the genome. Methylenetetrahydrofolate reductase (MTHFR) is a key regulatory enzyme involved in folate metabolism. Polymorphisms of MTHFR may change the level of homocysteine and affect DNA synthesis and methylation, leading to an increased oxidative stress and disturbed methylation reactions and consequently affecting reproductive function. This article presents an overview on MTHFR gene polymorphisms, proposing that multicentered, large-sample and long-term prospective studies are needed to reveal the relationship between MTHFR gene polymorphisms and infertility.
DNA ; biosynthesis ; DNA Methylation ; DNA Repair ; Folic Acid ; metabolism ; Homocysteine ; metabolism ; Humans ; Infertility ; enzymology ; genetics ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Genetic ; Prospective Studies

Animals ; Lung ; drug effects ; metabolism ; Perchlorates ; toxicity ; Quaternary Ammonium Compounds ; toxicity ; RNA, Messenger ; genetics ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

Objective To explore the effects of iPS cells-derived chimeric thymus transplantation on T cells reconstitution and graft versus host disease of murine after allo-BMT. Methods iPS cells-derived chimeric thymus was grafted under the renal capsules of mice after allogeneic IBM-BMT. The mice were divided into three groups:IBM-BMT group, IBM-BMT+TT group and IBM-BMT+DLI group. Four weeks after BMT, T lymphocyte subsets in the peripheral blood were analyzed by flow cytometry, the degree and pathological examination of GVHD were observed, respectively. Results Percentage of CD8+T cells in IBM-BMT group, IBM-BMT+TT group and IBM-BMT+DLI group was(5.52 ± 0.83)%,(11.10 ± 1.49)%and(8.49 ± 0.82)%respectively, there was signifi-cant difference between pairwise comparisons(P<0.05), and percentage of CD4 + T cells of the peripheral blood in IBM-BMT+TT group(9.60 ± 0.69)%was significantly higher than IBM-BMT group(6.42 ± 1.40)%and IBM-BMT+DLI group(8.07 ± 0.65)%(P<0.05) . IBM-BMT group and IBM-BMT+TT group showed less clinical and histopathological scoring of GVHD than IBM-BMT + DLI group. Conclusion iPS cells-derived chimeric thymus transplantation could effectively accelerate T cells reconstitution and prevent GVHD after allo-BMT.

Objective To examine an in vivo method for the differentiation of induced pluripotent stem cells (iPSCs) into thymic epithelial cells (TECs) in mice. Methods Green fluorescent protein-expressing iPS cells, derived from C57BL/6 mice, were injected into blastocysts from ICR mice. Chimeric blastocysts were then transferred into uteri of E2.5 pseudopregnant mice. Chimeric mouse could be identified by coat color 10 days after birth. The chimeric thymus was transplanted under the renal capsule of BALB/c nude mice. The spleen was cut out from the thymus-transplanted nude mice and the cells were dispersed and analyzed by a flow cytometer 4 weeks after transplantation. Results Chimeras were born 17 days after embryo transfer and 13 live-born chimeras were obtained. The contribution of iPSC-derived cells in the chimeras ranged from 5% to at most 90%. Typical thymic epithelium structure consisted of green fluorescent protein-expressing cells in chimera. The iPSCs-derived thymic epithelial cells could support the generation of new T cells. Conclusion The results indicate that mouse iPS cells can differentiate in vivo towards normally functioning TECs.

Objective:To explore the effect of extracorporeal shock wave combined with meloxicam tablets on joint function and serum matrix metalloproteinase-9 (MMP-9) and resistin levels in elderly patients with osteoarthritis.Methods:80 cases of elderly patients with osteoarthritis in Nanchong Central Hospital from May 2016 to March 2019 were selected as the research objects, and they were randomly divided into two groups, 40 cases in each group. The control group was given conventional oral medicine (meloxicam + potassium glucosamine sulfate tablets), and the observation group was given meloxicam tablets combined with extracorporeal shock wave therapy. The total effective rate, incidence of adverse reactions, and the scores of osteoarthritis symptoms (WOMCA), joint function (Lysholm), serum MMP-9 and resistin before and after 8 weeks' treatment were compared between the two groups.Results:After treatment, the WOMCA score of the observation group was lower than that of the control group, and Lysholm score was higher than that of the control group ( P<0.05); the total effective rate of the observation group (95.00%) was higher than of the control group (77.50%) ( P<0.05); the serum resistin and MMP-9 levels of the observation group were lower than those of the control group after treatment ( P<0.05); there was no significant difference in the incidence of adverse reactions between the observation group (15.00%) and the control group (17.50%) ( P>0.05). Conclusions:Extracorporeal shock wave combined with meloxicam tablets in the treatment of elderly patients with osteoarthritis can significantly improve the clinical symptoms, promote the recovery of joint function, reduce the levels of serum resistin and MMP-9, and further improve the treatment effect with high safety.