Cyclodextrin glucanotransferase, the essential enzyme for the production of cyclodextrins, has become the focus of scientific research nowadays. Although many related enzyme properties are well known, the crucial factors in product specificity determination remain to be answered. Here, the recent research progresses of cyclodextrin glucanotransferase, especially those about the evolution of product specificity, were reviewed, and the scientific problems were discussed.
Archaeal Proteins ; genetics ; metabolism ; Bacillus ; enzymology ; genetics ; Bacterial Proteins ; genetics ; metabolism ; Biocatalysis ; Cyclodextrins ; metabolism ; Evolution, Molecular ; Glucosyltransferases ; classification ; genetics ; metabolism ; Mutation ; Thermoanaerobacterium ; enzymology ; genetics ; Thermococcus

Recombination plasmid pMM085 possessed both immunogens heat-labile enterotoxin(LT) and fimbriae antigen K88 of enterotoxigenic Escherichia coli (ETEC). Althouth vaccine strain MM-3 carrying pMM085 had good effect to protect piglets against diarrhea due to ETEC infections,it was not ideal live vaccine for pMM085 bringing chloramphenicol resistance gene (cat). To solve the problem,the host-plasmid balanced lethal system was introduced which including the replacement of cat gene by asd gene and transformation the new plasmid to the strain X6097 which asd gene was knocked out in its chromosome. Considering pMM085 was a big plasmid (23kb) and traditional genetic manipulations was not easy to carry on,?-Red recombination system was adopt in this work to realize the replacement of cat gene by asd gene. The results indicated that ?-Red recombination system was convenient and efficient to reconstruct big plasmid.

Adult ; Aged ; Bone Plates ; Bone Transplantation ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Spinal Canal ; surgery ; Spinal Fusion ; Spinal Stenosis ; surgery ; Spondylolisthesis ; surgery

OBJECTIVETo investigate the effect of omega-3 polyunsaturated fatty acids(omega-3PUFAs) on the apoptosis of human gastric cancer cell line SGC-7901 and to explore the potential mechanisms.

METHODSCells were treated with eicosapentaenoic acid (20:5 omega-3,EPA) or docosahexaenoic acid (22:6 omega-3, DHA) at concentrations of 10, 20 and 40 microg/ml. Cell growth and apoptosis were analyzed with MTT assay, cell morphology, DNA electrophoresis and flow cytometry. Mitochondrial membrane potential ( triangle right psi mt) was measured by fluorescent probe rhodamine 123. The distribution of cytochrome C in mitochondria and cytosol was determined by enzyme-linked immunoadsorbent assay. The composition of mitochondrial membrane phospholipid(MMP)was examined by gas chromatography.

RESULTSBoth EPA and DHA markedly inhibited the SGC-7901 cell growth and induced apoptosis in a time- and dose-dependent manner. After incubation of the cells with 40 microg/ml EPA or DHA for 24 hours, the level of Deltapsimt siginificantly decreased (P<0.001), and cytochrome C largely released into cytosol from mitochondria. The proportions of EPA and DHA in MMP rapidly elevated while that of arachidonic acid sharply decreased.

CONCLUSIONSomega-3PUFAs inhibit the growth of gastric cancer cells through promoting apoptosis. Compositional and functional alterations in mitochondrial membrane may be an important initiator of apoptosis induced by omega-3PUFAs.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cytochromes c ; metabolism ; Fatty Acids, Omega-3 ; pharmacology ; Humans ; Membrane Potential, Mitochondrial ; Mitochondria ; metabolism ; pathology ; Stomach Neoplasms ; metabolism

OBJECTIVETo observe the effect of Migu capsule and Strengthening Spleen prescriptions on the expression of vitamin D receptor on small intestine and calf muscle of the rat, while observing whether the Chinese medicine complex prescriptions of different effect such as invigorating the kidney and strengthening the spleen had selectivity to expression of the VDR mRNA.

METHODSCopy the model of osteoporosis by ovariectomy operation, the rats were randomly divided into four groups, pseudo-resection group, model group, Migu capsule group and strengthening Spleen prescriptions group. Drug delivery 12 weeks later from operation, stomach lavage last for 12 weeks. Then, to observe the effect of medicine on the rats' bone mineral density, uterus weight, calf muscle/weight, and the VDR mRNA in small intestine and calf muscle through RT-PCR.

RESULTSMigu capsule enhanced bone mineral density (P<0.05), raised calf muscle/weight (P<0.05), up-regulated expression of calf muscle VDR mRNA in the small intestine (P<0.01). Strengthening Spleen prescriptions remained bone mineral density (P>0.05), up-regulated expression of calf muscle VDR mRNA (P<0.01), there was no obvious difference in uterus weight in Migu capsule group and Strengthening Spleen prescription group.

CONCLUSIONMigu capsule and Strengthening Spleen prescriptions can cure osteoporosis by improving the expression of calf muscle VDR mRNA in the small intestine.

Animals ; Bone Density ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation ; drug effects ; Intestine, Small ; drug effects ; metabolism ; Muscle, Skeletal ; drug effects ; metabolism ; Osteoporosis ; metabolism ; prevention & control ; Ovariectomy ; Prescriptions ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Calcitriol ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Spleen ; drug effects ; metabolism

Animals ; Antigens, CD ; immunology ; Cytokines ; metabolism ; Hepacivirus ; immunology ; Hepatitis C ; immunology ; prevention & control ; virology ; Hepatitis C Antibodies ; biosynthesis ; immunology ; Hepatitis C Antigens ; immunology ; Humans ; Killer Cells, Natural ; immunology ; T-Lymphocytes ; immunology ; metabolism ; T-Lymphocytes, Regulatory ; immunology ; metabolism ; Viral Proteins ; immunology

Adenine ; administration & dosage ; analogs & derivatives ; therapeutic use ; Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Biomarkers ; blood ; Critical Illness ; Cytokines ; blood ; DNA, Viral ; blood ; Follow-Up Studies ; Hepatitis B virus ; drug effects ; immunology ; Hepatitis B, Chronic ; blood ; drug therapy ; immunology ; virology ; Humans ; Lymphocyte Count ; Male ; Organophosphonates ; administration & dosage ; therapeutic use ; Recurrence ; T-Lymphocytes ; cytology ; immunology ; Time Factors ; Treatment Outcome

OBJECTIVETo investigate the status of male reproductive health among middle-aged and older men in the urban area of Nanjing.

METHODSWe collected the laboratory results of 884 middle-aged and older men aged 55 - 89 years from the Xuanwu District of Nanjing present for routine physical examinations, including those of blood routine tests, liver and kidney function, blood glucose, blood lipid, and total prostate specific antigen (TPSA), as well as such reproductive hormone indexes as total serum testosterone (TT), free serum testosterone (fT), and sex hormone binding globulin (SHBG). We also obtained the above reproductive hormone indexes from 119 young and middle-aged men aged 20 - 39 years as controls.

RESULTSAging-related changes were found in the 50 percentiles of all the reproductive hormones and relevant parameters but those of TT and E2, with gradual increases in LH, FSH and SHBG and decreases in fT, TSI and fTI. Comparison of reproductive hormones and relevant parameters by Mann-Whitney U test did not show any statistically significant differences in the TT level between any two of the five age groups (20 - 39, 55 - 59, 60 - 69, 70 - 79, and > or = 80 yr) (P > 0.05) except between the control and > or = 80 yr groups and the 60 - 69 and > or = 80 yr groups (P < 0.05), nor in the E2 level between any two groups, nor in the levels of LH and FSH except between the 55 - 59 and 60 - 69 yr groups and the 70 - 79 and > or = 80 yr groups, and nor in the levels of fT and TSI except between the 55 - 59 and 60 - 69 yr groups. However, there were significant differences in the levels of SHBG and fTI between any two age groups. Spearman correlation analysis revealed that fT, TSI, and fTI were correlated negatively with aging and LH (P < 0.05, I r I > 0.5) but weakly positively with cholesterol, blood glucose and hemoglobin (P < 0.05, /r/ < 0.5), SHBG and LH positively with aging, SHBG weakly negatively with blood glucose and hemoglobin, LH weakly negatively with hemoglobin, and TT weakly negatively with aging but positively with hemoglobin.

CONCLUSIONThe levels of serum testosterone, particularly that of fT, declined with aging in middle-aged and older men in the urban area of Nanjing, which may contribute to abnormal lipid metabolism, low hemoglobin and high blood glucose.

Aged ; Aged, 80 and over ; Aging ; blood ; Blood Glucose ; analysis ; Case-Control Studies ; China ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Prostate-Specific Antigen ; blood ; Sex Hormone-Binding Globulin ; analysis ; Statistics, Nonparametric ; Testosterone ; blood

OBJECTIVETo establish the human osteoblasts culture system in vitro, observe the effects of icariin on human osteoblasts proliferation and expression of OPG protein, and to explore the mechanism of promoting bone formation about human osteoblast in icariin.

METHODSThe femoral cancellous bone pieces were obtained from the operation. The enzyme digestion method was used for culturing. The third passage of human osteoblast was taken for experiments. The cells were divided into four groups, the control group was treated with 15% NCS-DMEM-F12 (1:1), the experimental groups were respectively treated with 10(-6), 10(-8), 10(-10) mol/L icariin. The MTT method was used to observe the proliferation of human osteoblast on 1, 3, 5, 7, 9 d; in 8, 10, 12 d, western blot was used to determine the expression of OPG protein on human osteoblast.

RESULTS1) Results of MTT: the icariin promoted the proliferation of human osteoblast. There was a concentration-response relation,while with the concentration of icariin increased, the ability was more obvious. There was statistically difference between 10(-6) mol/L icariin group (0.402 +/- 0.033) and the control group (0.268 +/- 0.031) (P<0.05). In the timely research, as the time prolong, the number of human osteoblast were more. At the fifth day, the human osteoblast entered rapid growth period, and access the growth platform stage; the icariin began to promote the proliferation of human osteoblast from the fifth day, which almost maintained to the 7th day and the 9th day, and most obvious in the 9th day. There was statistically difference between 10(-6) mol/L icariin group (0.402 +/- 0.033) and the control group (0.268 +/- 0.031) at the 9th day. (The results of OPG protein expression: in the control group, the expression of OPG protein was detected at the 8th day (1.01 +/- 0.08), and reached the expression peak (1.80 +/- 0.10), there was statistically different (P<0.05). In the different days and different concentration icariin groups, the expressions of OPG protein were all inferior to the control group. While the concentration decreasing, the expression was less. There were statistically difference (P<0.05). At the day 12, there was no significant difference of OPG protein expression between the 10(-6) mol/L icariin group and the 10(-8) mol/L icariin group (P>0.05).

CONCLUSIONThe effect of icariin promoting the proliferation of human osteoblast maybe is one of the mechanisms of improving the bone formation on human osteoblast.

Animals ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Flavonoids ; pharmacology ; Gene Expression Regulation ; drug effects ; Humans ; Osteoblasts ; cytology ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Osteoprotegerin ; metabolism

Objective To explore the impact of chronic complications of type 2 diabetic mellitus (T2DM)on quality of life(QOL)among T2DM outpatients in urban China.Methods A cross.Sectional study was carried Out in 1524 T2DM outpatients recruited from 15 hospitals in 4 major cities of China.Questionnaire interviews were used to collect data on general characters and complications of T2DM.SF-36 questionnaire(version 1)was used and self-completed by patients under the help from interviewers for those having difficulties in reading.T test,one-way ANOVA and generalized linear model were used to explore the association between complications of T2DM and quality of life.Results Complication Was a significant predictor leading to poorer SF-36 subscale scores(decreased by 4.68-16.06 scores)and two summary scores (PCS,MCS).The scores of role-physical subscale(decreased by 22.45％)and role-emotionsl subscale(decreased by 16.28％)decreased much more than the other subscales in patients with complication.Overall,QOI Was reduced by 11.02％in patients with complication.The QoL had a gradual decrease with the increasing number of complications.The average scores descended from 2.82 to 10.33 in the eight subscales with the increase of one complication.Conclusion Having complication or multiple complications seemed to be predietors of poor QOL among T2DM outpatients.