Organic solvent tolerant microorganism(OSTM) is a novel extremophile and it hasn't been systematically studied until 1980s.Relying on certain mechanisms,the OSTM is able to effectively de-fend and decrease the toxicity from organic solvents,which enable the OSTM to be potentially applied in the industrial fields such as whole-cell catalysis and environmental treatment,etc.The comprehen-sively understanding of the mechanisms involved in organic solvent tolerance of OSTM could be com-bined with genetic engineering in order to modify and optimize the various specifications of OSTM,and further broaden its application in other industrial areas.Latest studies on the tolerant mechanisms of OSTM,in this paper,will be reviewed from four aspects such as vesicle exocytosis and changes of phos-pholipid composition in membrane,etc.Besides,the application of OSTM in whole-cell catalysis and other fields will be introduced.

Humans ; Siblings ; Forensic Medicine ; Forensic Genetics

OBJECTIVETo investigate the effect of moxibustion at Sanyinjiao (SP 6) for uterine contraction pain in labor, and evaluate the safety of the parturient and newborn.

METHODSOne hundred and seventy-four cases of singleton pregnancy and cephalic presentation primipara were single blinded and randomly divided into three groups: observation group (59 cases), placebo treated group (57 cases) and blank group (58 cases). The observation group was treated with moxibustion at Sanyinjiao (SP 6) for 30 min when the uterus cervix openning at 3 cm, the placebo treated group was treated with moxibustion at no acupoint for 30 min and the blank group was treated with routine labor nursing, the uterine contraction pain and the safety of the mother and infant were compared among three groups.

RESULTS1) The uterine contraction pain was tested by Visual Analogue Scale (VAS): the scores of VAS in the observation group were obviously decreased after 15 min and 30 min of moxibustion (both P<0.05), there were no obvious changes of the VAS scores in placebo treated group and the blank group, the scores of VAS in observation group decreased much more obviously than those in the other two groups (all P<0.05); 2) Midwife rating of the uterine contraction pain: after 30 min of moxibustion, the effective rate of labor analgesia was 69.5% (41/59) in observation group, which was higher than that of 45.6% (26/57) in placebo treated group and 43.1% (25/58) in blank group, with significant differences between them (both P<0.05); 3) The postpartum hemorrhage amount of the observation group was obviously lower than those of placebo treated group and blank group (both P<0.05); 4) The Apgar score of newborn was higher in observation group and placebo treated group than that of blank group (both P<0.05).

CONCLUSIONMoxibustion at Sanyinjiao (SP 6) can relieve the uterine contraction pain, and has no side effect to mother and infant, it is one of the safe, effective and simple non-drug analgesia methods.

Acupuncture Points ; Adult ; Female ; Humans ; Labor Pain ; physiopathology ; therapy ; Labor, Obstetric ; Moxibustion ; Pregnancy ; Treatment Outcome ; Uterine Contraction ; Young Adult

AIM: To analyze the effect of calcium dobesilate combined with hypoglycemics for patients with diabetic retinopathy (DR) and cataract, and the influence on microcirculation of eye fundus, hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) level. METHODS: Totally 98 DR patients with cataract (126 eyes) from January 2013 to December 2015 in our hospital were selected as the research objects, and were divided into two groups randomly(treatment group:64 eyes in 49 patients, control group: 62 eyes in 49 patients). The control group was treated with acarbose tablet and metformin, while treatment group was treated with calcium dobesilate additionally. The clinical effect, the glycemic control effect, serum HIF-1α and VEGF level, eye function and fundus microcirculation of two groups after 12mo were compared. RESULTS: After 12mo, the total effective rates of two groups were 87.5%,61.3% respectively, which indicated significantly difference (P<0.05); the vision of treatment group was significantly higher than that of control group (P<0.05). Two groups' blood glucose level decreased significantly, and no significant difference between two groups(P>0.05). After 1-month treatment, the plasma viscosity, erythrocyte aggregation index, erythrocyte deformability index, erythrocyte sedimentation rate and hematocrit in the treatment group were significantly lower than those in the control group (P<0.05). The PSV and EDV of the posterior ciliary artery and central artery in the treatment group were significantly higher than those in the control group (P<0. 05), and RI was significantly lower than that in the control group (P<0.05). After 12-month treatment, the HIF-1α level of two groups were 35 90士11.36mmol/L, 46.75士12.08mmol/L respectively;the VEGF of two groups level were 89.72士13.61mmol/L, 110.30士16.74mmol/L, respectively, the treatment group's HIF- 1α level and VEGF were significantly lower than control group (P<0.05), and both decreased significantly after treatment (P<0.05). CONCLUSION: Calcium dobesilate combined with hypoglycemics can effectively increase the clinical effect in the treatment of retinopathy diabetic cataract, effectively control blood glucose, improve microcirculation of eye fundus, decrease HIF - 1α, VEGF level, inhibit angiogenesis.

Hepacivirus ; isolation & purification ; Humans ; Leukocytes, Mononuclear ; virology ; RNA, Viral ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viremia ; virology

OBJECTIVETo compare the number of the Tannerella forsythensis (T. forsythensis), total bacteria, and proportion of T. forsythensis in subgingival specimens in diseased sites of chronic periodontitis patients and in healthy sites of periodontally healthy subjects, and clarify the relationship between bacterial load and periodontal status.

METHODSSubgingival plaque samples from 61 chronic periodontitis patients and 12 healthy controls (positive for T. forsythensis by conventional PCR) were analyzed with TaqMan real-time polymerase chain reaction assay for T. forsythensis and total bacteria. Quantification was performed with species-specific primer/probe, universal primer/ probe and serial dilution of plasmid standards.

RESULTSNumbers of T. forsythensis and total bacteria(P<0.001) , the proportion of T. forsythensis in subgingival specimens (P<0.05) were significant higher in diseased sites of chronic periodontitis patients than in healthy sites of healthy subjects. In addition, a significant correlation was found between the number of bacteria and various probing depth (P<0.001). There was no significantly difference between the proportion of T. forsythensis in subgingival plaque and probing depth.

CONCLUSIONNumber of T. forsythensis are closely associated with periodontal status, and demonstrate the broad potential of real-time polymerase chain reaction application on periodontology.

Adult ; Bacteroides ; Chronic Periodontitis ; Dental Plaque ; Female ; Humans ; Male ; Middle Aged ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas gingivalis

OBJECTIVETo construct a plasmid vector expressing the short hairpin RNA (shRNA) targeting proliferation cell nuclear antigen (PCNA), and to investigate its effect in vitro on the expression of PCNA, proliferation and apoptosis of human osteosarcoma cells.

METHODSA plasmid vector expressing the short hairpin RNA targeting at PCNA was constructed and transfected into human osteosarcoma cell line MG-63 by dosper liposomal method. PCNA mRNA and protein expressions were examined using RT-PCR and immunohistochemical staining, respectively. Inhibition of the cell proliferation was studied by MTT method and colony forming assay. DNA synthesis was analyzed by (3)H-TdR incorporation and the cell cycle was determined by flow cytometry. The apoptotic cells were stained with acridine orange.

RESULTSExpression of PCNA mRNA after the transfection was markedly inhibited by 80.51% with a PI value of 25.68% of that of the control group. PCNA shRNA inhibited MG-63 cell growth detected by using MTT method, with an inhibition rate of 61.78% at 48 h. DNA synthesis rate also decreased in the (3)H-TdR incorporation test. Flow cytometry analysis showed an increase of the percentage of G(0)/G(1) phase cells, along with a decrease of cell population in the S phase. The apoptosis rate of cells transfected with the plasmid vector was 16.54%.

CONCLUSIONSPCNA shRNA significantly suppresses the expression of PCNA at both mRNA and protein levels, corresponding to an inhibition of the proliferation of MG-63 cell and an increase of the cellular apoptosis.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Osteosarcoma ; metabolism ; pathology ; Plasmids ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; biosynthesis ; genetics ; Transfection

OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

BACKGROUNDThis study was to establish a disease differentiation model for ST-segment elevation myocardial infarction (STEMI) youth patients experiencing ischemia and reperfusion via ultra-performance liquid chromatography and mass spectrometry (UPLC/MS) platform, which searches for closely related characteristic metabolites and metabolic pathways to evaluate their predictive value in the prognosis after discharge.

METHODSForty-seven consecutive STEMI patients (23 patients under 45 years of age, referred to here as "youth," and 24 "elderly" patients) and 48 healthy control group members (24 youth, 24 elderly) were registered prospectively. The youth patients were required to provide a second blood draw during a follow-up visit one year after morbidity (n = 22, one lost). Characteristic metabolites and relative metabolic pathways were screened via UPLC/MS platform base on the Kyoto encyclopedia of genes and genomes (KEGG) and Human Metabolome Database. Receiver operating characteristic (ROC) curves were drawn to evaluate the predictive value of characteristic metabolites in the prognosis after discharge.

RESULTSWe successfully established an orthogonal partial least squares discriminated analysis model (R2X = 71.2%, R2Y = 79.6%, and Q2 = 55.9%) and screened out 24 ions; the sphingolipid metabolism pathway showed the most drastic change. The ROC curve analysis showed that ceramide [Cer(d18:0/16:0), Cer(t18:0/12:0)] and sphinganine in the sphingolipid pathway have high sensitivity and specificity on the prognosis related to major adverse cardiovascular events after youth patients were discharged. The area under curve (AUC) was 0.671, 0.750, and 0.711, respectively. A follow-up validation one year after morbidity showed corresponding AUC of 0.778, 0.833, and 0.806.

CONCLUSIONSBy analyzing the plasma metabolism of myocardial infarction patients, we successfully established a model that can distinguish two different factors simultaneously: pathological conditions and age. Sphingolipid metabolism is the top most altered pathway in young STEMI patients and as such may represent a valuable prognostic factor and potential therapeutic target.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Humans ; Least-Squares Analysis ; Mass Spectrometry ; methods ; Myocardial Ischemia ; metabolism ; Myocardial Reperfusion ; ST Elevation Myocardial Infarction ; metabolism ; Sphingolipids ; metabolism

OBJECTIVETo investigate the expression of the lysosomal enzyme acid sphingomyelinase (ASMase) in alcohol-induced hepatic fibrosis using a rat model.

METHODSThe model of liver fibrosis was induced by administration of alcohol and high fat diet using 20 rats. Six rats given no alcohol and normal diet served as the control group. Real-time PCR, western blotting, and immunohistochemistry were used to evaluate fibrosis-related changes in the mRNA and protein expressions of ASMase.

RESULTSThe fibrotic liver tissues of the model rats showed significantly higher expression levels of ASMase than the non-fibrotic liver tissues of the control rats (P less than 0.05).

CONCLUSIONExpression of ASMase is increased in the fibrotic liver tissue of an alcohol-induced hepatic fibrosis rat model, suggesting that this lysosomal enzyme may contribute to development of this disease condition.

Animals ; Liver ; enzymology ; Liver Cirrhosis, Alcoholic ; enzymology ; Liver Cirrhosis, Experimental ; enzymology ; Male ; Rats ; Rats, Sprague-Dawley ; Sphingomyelin Phosphodiesterase ; metabolism