Chromosomally encoded toxin–antitoxin (TA) systems are thought to result in growth arrest and eventual cell death upon exposure to environmental stress in E. coli. In the chromosome of cyanobacteria Synechocystis sp. PCC6803, the genetic organization of a 360 bp open reading frame (ORF), slr0664, and another small ORF of 256 bp, ssr1114, is similar to that of TA system. The predicted protein encoded by slr0664 is homologous to RelE, but neither homologue of ssr1114 nor ssr1114-encoding protein was found in TA system. To see whether slr0664 encodes a toxin protein, ssr1114 encodes an antitoxin, an expressing plasmid containing promoter Plac and PBAD, was constructed. In this construct, Both slr0664 and ssr1114 were controlled by Plac and PBAD, respectively. Expression of slr0664 in Escherichia coli results in the inhi-bition of bacterial growth, the expression of ssr1114 neutralize the toxicity of slr0664 expression. These re-sults show that slr0664 is toxin gene and ssr1114 is antitoxin gene, both ssr1114 and slr0664 constitute achromosomal TA system in Synechocystis sp. PCC6803.

Objective To observe bone formation of compound of autologous periosteum cells plus calcium alginate gelatin in animal body and investigate the best mixing ratio of the two components.Methods A total of 96 healthy adult New Zealand rabbits were randomly divided into six groups (Groups A,B,C,D,E and control group).The excised bilateral periostea from rabbits in each experimental group were all made into 0.4 ml cell suspensions.The compounds were prepared by blending 0.4 ml calcium alginate gelatin with 1,1/4,1/16,1/64 and 1/256 of the periosteum cell suspension respectively,and then were applied to autologous right radial defect area.Gross observation,X-ray examination and histological study were carried out at 2,4,8,12 weeks postoperatively.Levels of serum alkaline phosphatase and serum calcium were determined as well.Results The compounds containing periosteum cell suspension with cell counts of 1/4 or 1/16 and calcium alginate gelatin reached the best osteogenesis.Conclusion Compound of autologous periosteum cell suspension-calcium alginate gelatin induces obvious bone formation and is worthy of clinical application,for it has advantages of satisfactory bone defect repair and easy operation.

OBJECTIVE:To prepare diclofenac sodium controlled release pellets and investigate the mechanism of re?lease.METHODS:Single-factor method was used to investigate the influence of the composition of solvent system in coating solution,the concentration of coating material,porogenic agent and plastifier on the release rate of drug.RESULTS:When the concentration of coating material and the proportion of water increased,the release rate of pellets was increased;the release rate was further increased with PVP K30 added.CONCLUSION:The pellets belongs to the matrix-coating pellets,and the release mechanism varies with the change of coating thickness.

Objective To compare the saccharometabolism with the pancreatic islets functions and insulin resistance index in children with severe stress. Methods Thirty children with severe stress and 30 healthy children in control group were tested. The levels of fasting blood glucose (FBG), fasting insulin (FINS) and fasting C - peptide (FCP) were detected by radioimmunoassay respectively and insulin sensitivity index (ISI), insulin resistance index (IR) and fasting blood cell function index (FBCI) were calculated statistically. Results There were significant differences between the children with severe stress and the normal controls in the levels of FINS, FCP and FBG,(all P0.05). Conclusion There is insulin resistance with the significant decrease in the insulin sensitivity index and significant increase in insulin resistance index in the children with severe stress, which may cause the disorder in glucose metabolism in children with severe stress.

The growth changes of glutathione (GSH) and ergosterol in Saccharomyces cerevisiae (CICC1447 and CICC1339) were detected under 0.5Mpa pressure with compressed high-pure air (O-2∶N-2=21∶79). The results showed that logarithmic phases of the two strains were delayed; their biomass and special growth rate were lower than those of control sample (0.1MPa) and the double time were prolonged under 0.5MPa. High-pressure could increase the content of GSH obviously, compared to ambient atmosphere control samples. When the holding time was 3h, the content of GSH and ergosterol in CICC1447 increased 42.6% and 20.1%, respectively. However, the content of GSH in CICC1339 increased 58.7% when the holding time was 6h, while ergosterol content reduced. The results indicated that different yeast strains have different stress-response mechanism to copy with high-pressure shock.

OBJECTIVETo summarize the features and treatment of male infertility induced by autosomal dominant polycystic kidney disease (ADPKD), and compare the outcomes of intracytoplasmic sperm injection (ICSI) for infertile men with ADPKD and those with congenital bilateral absence of vas deferens (CBAVD).

METHODSWe retrospectively analyzed 21 cases of ADPKD-induced infertility, 15 treated by ICSI (group A), and another 164 cases of strictly matched CBAVD-induced infertility (group B). We compared the two groups in the couples' age, the number of ICSI oocytes, and the rates of fertilization, transferrable embryos, good embryos, embryos implanted, clinical pregnancy, biochemical pregnancy, early abortion, singleton and twins in the first cycle.

RESULTSAfter 28 cycles of ICSI, 10 of the 15 ADPKD-induced infertility patients achieved clinical pregnancy, including 7 cases of live birth, 1 case of spontaneous abortion, and 2 cases of pregnancy maintenance. No significant differences were observed between groups A and B in the couples' age, the wives' BMI, or the numbers of ICSI oocytes and embryos transplanted (P >0.05), nor in the rates of ICSI fertilization (72.64% vs 76.17%), transferrable embryos (51.28% vs 63.24%), quality embryos (38.46% vs 49.83%), embryo implantation (17.64% vs 38.50%), abortion (0 vs 9.23%), singleton (50% vs 81.54%) and twins (50% vs 18.46%). However, the rates of clinical pregnancy (13.33% vs 42.68%, P = 0.023 <0.05) and biochemical pregnancy (13.33% vs 39.63%, P = 0.032 <0.05) were significantly lower in group A than in B.

CONCLUSIONICSI is effective in the treatment of male infertility induced by either ADPKD or CBAVD, but the ADPKD cases have a lower success rate than the CBAVD cases in an individual cycle. The affected couples should be informed of the necessity of prenatal genetic diagnosis before embryo implantation and the inevitable vertical transmission of genetic problems to the offspring.

Abortion, Spontaneous ; Embryo Implantation ; Embryo Transfer ; Female ; Humans ; Infertility, Male ; therapy ; Male ; Male Urogenital Diseases ; therapy ; Oocytes ; Polycystic Kidney, Autosomal Dominant ; complications ; Pregnancy ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; Vas Deferens ; abnormalities

Pregnane X receptor (PXR), a member of nuclear receptor superfamily, plays an important role in xenobiotic and endogenous metabolism, endocrine balance, and cell proliferation, etc. Previous study has shown that pregnenolone 16α-carbonitrile (PCN), a mouse PXR agonist, could induce liver enlargement. And we found that the change in hepatocytes exhibits regional distribution characteristics: hepatocyte enlargement occurs around the central vein (CV) area, while hepatocyte proliferation occurs around the portal vein (PV) area. In this study, the dynamic changes of hepatocytes during PXR-induced liver enlargement were determined. Serum and liver samples from male C57BL/6 mice were collected for biochemical and pathological analysis after PCN treatment for 1, 2, 3, 5 days, respectively. The animal experiment was approved by the Animal Ethics Committee of Sun Yat-Sen University. The results showed that with the increase in the PCN treatment days, the feature of this regional change of hepatocyte around the CV and PV areas became more and more obvious. At the same time, the factors related to hepatocyte enlargement, such as the expression of PXR downstream genes and the hepatic content of triglyceride (TG), has gradually increased. The upregulation of proliferation-related proteins and downregulation of cyclin-dependent kinases inhibitor proteins were observed in the early stage of PCN treatment, suggesting that hepatocyte proliferation occurs earlier than hepatocyte enlargement during PXR-induced liver enlargement. This study reveals the dynamic change of hepatocytes during PXR-induced liver enlargement and provides a new insight in liver enlargement promoted via PXR activation.

Adolescent ; Diskectomy ; Female ; Humans ; Intervertebral Disc Displacement ; physiopathology ; surgery ; therapy ; Lumbar Vertebrae ; pathology ; physiopathology ; surgery ; Male ; Postoperative Period ; Time Factors ; Treatment Outcome ; Young Adult

This study aims to investigate the function of two SNPs (rs8904C > T and rs696G >A) in 3' untranslated region (3'UTR) of NFKBIA gene by constructing luciferase reporter gene. A patient's genomic DNA with rs8904 CC and rs696 GA genotype was used as the PCR template. Full-length 3'UTR of NFKBIA gene was amplified by different primers. After sequencing validation, these fragments were inserted to the luciferase reporter vector, pGL3-promoter to construct recombinant plasmids containing four kinds of haplotypes, pGL3-rs8904C/rs696G, pGL3-rs8904C/rs696A, pGL3-rs8904T/rs696G and pGL3-rs8904T/rs696A. Then these plasmids were transfected into LS174T cells and the luciferase activity was detected. Compared with pGL3-vector transfected cells (negative control), the luciferase activity of the four kinds of recombinant plasmids was significantly decreased (P < 0.001). For rs696G > A, the luciferase activity of the recombinant plasmids containing A allele (pGL3-rs8904C/rs696A and pGL3-rs8904T/rs696A) was about 45.1% (P < 0.05) and 56.1% (P < 0.001) lower than those containing G allele (pGL3-rs8904C/rs696G and pGL3-rs8904T/rs696G), respectively. For rs8904C > T, there were no significant differences in the luciferase activity between the recombinant plasmids containing T allele and those with C allele. Together, the luciferase reporter gene vectors containing SNPs in NFKBIA gene 3'UTR were constructed successfully and rs696G > A could decrease the luciferase activity while rs8904C >T didn't have much effect on the luciferase activity.
3' Untranslated Regions ; Genes, Reporter ; Genetic Vectors ; Humans ; I-kappa B Proteins ; genetics ; Luciferases ; NF-KappaB Inhibitor alpha ; Plasmids ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Transfection

Fifteen known compounds were isolated from Swertia delavayi by silica gel, Sephadex LH-20 and Rp-18 column chromatographies. Based on extensive spectroscopic analysis (MS, 1H, 13C-NMR), their structures were identified aserythrocentaurin (1), erythrocentaurindimethylacetal (2), sweroside (3), swertiamarin (4), gentiopicroside (5), swertiakoside A (6), 2'-O-acetylswertiamarin (7), 4'-O-[(Z) -coumaroyl] swertiamarin (8), 1,5,8-trihydroxy-3-methoxyxanthone (9), 8-O-β-D-glucopyranosyl-1-hydroxy-2,3, 5-trimethoxyxanthone (10), 8-O-[β-D-xyl- opyranosyl-(1 --> 6)-β-D-glucopyranosyl]-7,8-dihydroxy-3-methoxyxanthone (11), isovitexin (12), β-sitosterol (13), daucosterol (14), and oleanolic acid (15). Among them, ten ones (14, 7-11, 13) were obtained from S. delavayi for the first time. The isolates were evaluated for their anti-HBV activities in HepG 2. 2. 15 cell line in vitro. The results showed that compound 1, 2, 6, 7, 9 and 12 exhibited significant inhibitory activity on HBV DNA replication with IC50 values from 0.05 to 1.46 mmol x L(-1).
Antiviral Agents ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Hepatitis B virus ; drug effects ; genetics ; Magnetic Resonance Imaging ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Swertia ; chemistry