Biodiesel, a nontoxic,cleaning, renewable and biodegradable fuel, is expected as a substitute for conventional fossil diesel. There are three main approaches to produce biodiesel, alkali-catalysis processing, enzymatic-catalysis processing and supercritical processing. With the unique property of energy-saving and environment-friendly, enzymatic-catalysis appears a great potential for industrial application. The main bottleneck of this technology is high cost and low stability of the lipase, as well as the inactivation of lipase by methanol and so on. To settle the problem, several methods have been used including the fixed-bed bioreactor, enzyme immobilized processing, whole-cell biocatalyst, changing addition method of methanol, developing of novel acyl acceptor, enhancing methanol resistance of lipase. The main problems and the relative strategy research of the enzymatic-catalysis technology were sum up.

Objective:A novel tuberculosis DNA vaccine integrating siRNA,fusional antigen and hIL-12 triple expression units was constructed in our laboratory.Methods:To evaluate the independent expression of the three expression units,two eukaryotic expression plasmid pVAX1-siRNA-PVAE[EGFP]-hIL12 with TB fusional gene Ag85B-ESAT6(PVAE) and enhanced green fluorescent protein(EGFP),and pVAX1-siEGFP-PVAE[EGFP]-hIL12 with siRNA coding sequence targeting EGFP instead of Mcl-1L were constructed.Then two plasmids were used to transfer human embryonic kidney 293 cells.Based on EGFP report gene,it was demonstrated that siRNA expression unit and fusional antigen gene were independently expressed.Results:The hIL12 expression at 48h and 72h post transfection were also detected by ELISA analysis up to 1571.63pg/ml and 2392.25pg/ml respectively in the cell culture fluid.Conclusion:The results demonstrated that the novel DNA vaccine with siRNA,TB fusional antigen and hIL12 three expression units in the same plasmid frame is successfully constructed and independently expressed in eukaryotic cells.It laid a foundation for further animal model study on anti-tuberculosis effects of this novel DNA vaccine.

Objective To investigate the relationship between p62 expression,and occurrence and metastasis of nasopharyngeal carcinomas.Methods Immunohistochemical method was used to analyze p62 expression in 123 nasopharyngeal carcinoma (NPC) cases and 30 chronic nasopharyngitis cases.The clinical pathological characteristics were analyzed.Results The positive rate of p62 protein in chronic nasopharyngitis nasopharyngeal epithelium,non-metastatic NPC tissue,and metastatic NPC tissues was 13.3％,66.67％,and 84.72％,respectively,and the difference was statistically significant.The expression of p62 protein in nasopharyngeal carcinoma patients with lymph node metastasis or distant metastasis was significantly higher than non-metastatic NPC patients (P ＜ 0.01 or P ＜ 0.05).However,the expression of p62 was not related to age,gender,tumor size,and TNM stage (P ＞ 0.05).Conclusions High p62 protein expression in nasopharyngeal carcinoma tissue is closely related to the occurrence and metastasis of nasopharyngeal carcinomas.It provides good reference value to predict NPC malignancy and metastases.

Female ; Genetic Predisposition to Disease ; HSP70 Heat-Shock Proteins ; genetics ; Humans ; Lung Neoplasms ; genetics ; Male ; Polymorphism, Genetic

Objective To evaluate the possible molecular pathogenesis of intractable temporal lobe epilepsy. The potassium ion channel gene KCNJ4 encodes one of the subfamilies of Kir channels, Kir2.3 subunit, which may play an important role in modulating neuronal excitation. Interference in the function or expression of this gene would cause disturbance of ionic concentrations, thus leading to seizure activity. Methods Reverse transcription polymerase chain reaction (RT-PCR) and Western-blot analysis were used to measure the expression alterations of KCNJ4 mRNA as well as its protein product Kir2.3 channel in temporal cortex samples from patients who had undergone temporal lobectomy for intractable epilepsy (n=12). Tissue from 10 subjects who did not have epilepsy served as controls. Results The expression of KCNJ4 mRNA (0.438?0.178) and its protein Kir2.3 (M 50=0.063) were significantly decreased in epileptic brain compared with the controls (P

The extracting technology of salidroside, tyrosol, crenulatin and gallic acid from Rhodiolae Crenulatae Radix et Rhizoma was optimized. With extraction rate of salidroside, tyrosol, crenulatin and gallic acid as indexes, orthogonal test was used to evaluate effect of 4 factors on extracting technology, including concentration of solvent, the dosage of solvent, duration of extraction, and frequency of extraction. The results showed that, the best extracting technology was to extract in 70% alcohol with 8 times the weight of herbal medicine for 2 times, with 3 hours once. High extraction rate of salidroside, tyrosol, crenulatin and gallic acid were obtained with the present technology. The extracting technology was stable and feasible with high extraction rate of four compounds from Rhodiolae Crenulatae Radix et Rhizoma, it was suitable for industrial production.
Chemical Fractionation ; methods ; Chemistry, Pharmaceutical ; methods ; Coumarins ; isolation & purification ; Drugs, Chinese Herbal ; isolation & purification ; Gallic Acid ; isolation & purification ; Glucosides ; isolation & purification ; Phenols ; isolation & purification ; Phenylethyl Alcohol ; analogs & derivatives ; isolation & purification ; Rhizome ; chemistry ; Rhodiola ; chemistry

Isolated alkaloids from Coptis chinensis Franch. The compounds were identified as berberine, columbamine, groenlandicine, jatrorrhizine, magnoflorine, corydaldine and ferulic acid methylester. Then measured their bitter degree based on the electronic tongue and evaluated the antibacterial. The results based on the Electronic Tongue showed that berberine, columbamine, groenlandicine and jatrorrhizine have higher bitter degree than magnoflorine and corydaldine. And they also appeared better antibacterial activity on E. coli and S. aureus. The correlation coefficients between bitter degree and the two bacteria antibacterial activity were 0.983 and 0.911. So there was close relationship between the bitter degree and antibacterial activity of bitter components. Thus, it is confirmed further that bitter components are the material foundation of medicinal effectiveness of bitter herbs.
Aporphines ; analysis ; Berberine ; analogs & derivatives ; analysis ; Berberine Alkaloids ; analysis ; Biomedical Research ; instrumentation ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; analysis ; chemistry ; pharmacology ; Electronics ; instrumentation ; methods ; Escherichia coli ; drug effects ; growth & development ; Microbial Sensitivity Tests ; Reproducibility of Results ; Staphylococcus aureus ; drug effects ; growth & development ; Taste

Objective To investigate the effects of ciprofloxacin on anastomotic stoma,small intestinal inflammation and fibrosis in interleukin-10 knock-out mice (mice model of Crohn′s disease (CD)) after ileocaecal resection.Methods Twelve interleukin-10 knockout male C3H/HeJBir mice were evenly divided into control group and ciprofloxacin group.The mice model was established with ileocaecal resection and ileocolostomy.There was no intervention in control group.Ciprofloxacin (50 mg/kg per day)was orally administrated in ciprofloxacin group for four weeks.All the mice were executed.The intestine around anastomotic stoma was taken for hematoxylin-eosin (HE)and Sirius red staining and histological scoring.The level of interferon (IFN)-γand tumor growth factor (TGF)-βin intestine around anastomotic stoma and small intestine was detected by enzyme-linked immunosorbent assay (ELISA) method.The content of procollagen α1 was determined by real-time polymerase chain reaction.The data were analyzed by t test.Results The inflammatory reaction level of anastomotic stoma and small intestine in mice of ciprofloxacin group (2.50±0.55 and 1 .67±0.52)was significantly lower than those of control group (3.67±0.63 and 2.83±0.75 ,t=-8.273,-5 .188,both P <0.05 ).The IFN-γof anastomotic stoma and small intestine also significantly decreased ((0.72 ±0.22 )pg/mg vs (1 .76 ±0.43 )pg/mg, (0.23±0.12)pg/mg vs (0.78 ± 0.52 )pg/mg;t = - 7.354,- 5 .268,both P < 0.05).However, ciprofloxacin could not significantly decrease fibrosis in anastomotic stoma and small intestine of CD mice. There was no significant difference in the level of TGF-βcompared with control group.Conclusions The role of ciprofloxacin in improving inflammatory reation in anastomotic stoma of CD mice was limited. Maintenance therapy may be necessary to reduce the relapse of anastomotic stoma diseases.

Lipase gene from Penicillium expansum(lip07) was cloned and over-expressed in Pichia pastoris.a random mutant named ep8,which contained a single amino acid substitution,was obtained by using the lip07 as an error-prone PCR template in previous study.ep8 shows higher thermostability than that of lip07,To further improve the thermostability of the lipase,the Lys of wild-type(lip07) and mutant(ep8) in 202 were substituted by Ala using the Overlap extension PCR technique respectively.The mutant genes(lip07-K202A and ep8-K202A) were subcloned into pAO815,and then transformed into the Pichia pastoris GS115 for extracelluar expression,respectively.15% SDS-PAGE analysis indicated that the molecular mass of PEL-ep8-K202A and PEL-lip07-K202A are both about 28kDa,which is same with the wild-type lipase.The Tm of PEL-ep8-K202A is 41.66℃,2.63℃ higher than that of the wild-type(39.03℃) and 1.21℃ higher than the random mutant(PEL-ep8:40.45℃);the Tm of single mutant(PEL-lip07-K202A) is 37.08℃,2℃ lower than that of the wild-type lipase.

Aim The effects of antibiotics and anticoagulants on mouse peritonaeum were ob-served to explore the factor of the peritoneal dialysis related sclerosing peritoni-tis. Methods The experimental models of peritoneal dialysis were established in miceby infusing different kind of drugs to the peritoneal cavity and the changes of the peri-toneal membrane for each drug at different time were observed by the autopsy and lightmicroscope for several weeks. Results Amikacin, Cefradine, Zinacef, Ciprofloxacin,Heparin and Urokinase could induce sclerosing changes of peritoneal membrane such asloss of peritoneal mesothelum infiltration of inflammatory cells and of proliferation fibrecell.These changes were irreversible after the drugs were stoped.Conclusion Thedrugs commonly used in peritoneal dialysis may in different degree result in peritonealsclerosis.