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.

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.

In order to make the experimental teaching adapt to the development of modern teaching idea and to meet the needs of the pharmaceutical industry for high quality talents, pharma-cology experiment teaching method has been reformed. Single teaching method has turned to diversi-fied teaching method according to the experimental contents, difficulty and characteristics of teaching situation, such as leading method of using theory after experiment in validation experiments and single blind method in multidrug efficacy experiment in the early stage, case-based teaching in comprehensive experiments in the middle stage , and student teaching method in designing experiments in the later stage. The study results show that students' interest in learning has been inspired,their experimental enthusiasm has been mobilized,and their operation, analysis and problem-solving ability has been improved, which is advantageous to the comprehensive quality education.

Objective To verify whether the periodic or continuous exposure to high glucose may have different effects on human umbilical vein endothelial cell (HUVEC)apoptosis, and to explore the effect of NF-κB pathway on apoptosis of HUVEC induced by high glucose using the RNAi adenovirus vector. Methods RNAi combinant adenovirus vector which targeted 1566 site of NF-κB p65 mRNA was constructed and the effect of p65 gene knockdown in HUVEC was detected by Western blot analysis. Then, the RNAi adenovirus was transducted to explore the role of NF-κB pathway on the regulation of apoptosis in HUVEC induced by high glucose. The apoptosis of HUVEC was tested by flow cytometry and TUNEL assay. Results High glucose could induce apoptosis of HUVEC. p65 protein expression of nuclear extracts was significantly increased in high glucose culture as compared to control group, but only slightly increased in NF-κB-specific knockdown group, which maintained at basal state. Compared with normal glucose group, the number of TUNEL-positive cells in high glucose group was significantly increased (25.81%±1.77% vs 8.20%±0.63%, P<0.05). The number of TUNEL-positive cells was decreased in 30.5 rmnol/L glucose plus Ad-1566 than that in 30.5 mmol/L glucose plus Ad-DEST (11.49%±0.92% vs 26.10%±0.98%, P<0.01). Flow cytometry and TUNEL assay showed that the apoptosis of human umbilical vein endothelial cells induced by high glucose was inhibited by the RNAi adenovirus. Conclusion High glucose induces apoptosis of HUVEC. Knockdown of NF-κB p65 may protect HUVEC from apoptosis by preventing high glucose-induced NF-κB nuclear translocation.

OBJECTIVETo analyze and evaluate the acceptability of mifepristone compatible with misoprostone versus conventional surgical abortion among women under unwanted early pregnancy, so as to help the unexpected pregnant women to choose the satisfactory abortion, and to provide the evidence for clinicians to make a proper clinical decision.

METHODSSix medical databases were searched, including MEDLINE, EMBASE, Cochrane library, CBMdisc, CNKI and VIP, together with twelve journals hand-searched, and references of included studies additionally searched. Two qualified reviewers reviewed the original articles, evaluating qualities of articles, and extracting data independently. After heterogeneity test, the data was pooled using Revman software if capable, or descriptive analysis was applied.

RESULTSIn total, nine original clinical controlled trials were included, containing 3565 cases. Before abortion, more unwanted pregnant women chose the medical abortion because they believed medical abortion was less painful than surgical abortion (OR = 466.51, 95% CI: 91.37 - 2381.88), but medical abortion was less time-consuming than surgical abortion (OR = 0.02, 95% CI: 0.01 - 0.06). After abortion, satisfaction with medical abortion was similar to that with surgical abortion, with insignificant difference (P = 0.89). However, second choice and recommendation rates of medical abortion were much higher than those of surgical abortion with OR and 95% CI as 2.72, 2.13 - 3.47 and 4.19, 2.16 - 11.16, respectively.

CONCLUSIONSMedical abortion was less painful than surgical abortion and the rate of second choice and recommendation to others were all also higher than those of surgical abortion. However, the process of medical abortion was not as quick as surgical abortion but the satifacation of both methods seemed similar. Therefore, the two artificial abortion methods were not recommended to replace each other at the present time.

Abortifacient Agents ; adverse effects ; Abortion, Induced ; adverse effects ; methods ; Choice Behavior ; Controlled Clinical Trials as Topic ; Female ; Humans ; Mifepristone ; Misoprostol ; Pain ; etiology ; Patient Satisfaction ; statistics & numerical data ; Pregnancy ; Pregnancy, Unwanted

OBJECTIVETo study the epidemiological features of tic disorders (TD) among schoolchildren in Wenzhou area.

METHODSStratified cluster sampling was carried out to investigate TD in 9742 schoolchildren aged 7 to 16 years old in Wenzhou.

RESULTSThe average prevalence rate of TD among school-age children was 104/10 000 (166/10 000 for males, 29/10 000 for females). There was a significantly higher prevalence rate for males than that for females (chi(2) = 43.96, P < 0.001, prevalence ratio = 5.7, prevalence ratio 95% CI: 3.20 - 10.30). The prevalence rates of clinical subtypes in males was significantly higher than that of females while pupils was significantly higher than that in high school students (chi(2) = 11.33, P < 0.01, prevalence ratio = 2.2, prevalence ratio 95% CI: 1.37 - 3.43). Prevalence rate of transient tic disorders (TTD), chronic motor vocal tic disorder (CMVTD), tourette syndrome (TS) were 34/10 000, 27/10 000 and 43/10 000 respectively with the highest among 9-10 years old group. The mean onset age of TD was 8.5 +/- 2.8 years. The peak of onset was among 6-10 year olds. The rate of delayed diagnosis of the disorders was 69.3% and the median in delayed diagnosis was 1.0 year.

CONCLUSIONTD is a common disease with high rate of misdiagnoses among schoolchildren in Wenzhou area. Physicians and population should be trained to identify the syndromes and to practice correct diagnosis and effective treatment as early as possible.

Adolescent ; Child ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Prevalence ; Sex Factors ; Tic Disorders ; epidemiology ; prevention & control ; Tourette Syndrome ; epidemiology ; prevention & control

OBJECTIVETo study the prevalence and distribution of Tourette syndrome (TS).

METHODSNine thousand, seven hundred and forty-two children and juveniles aged 7 - 16 years in Wenzhou were studied, using cluster random sampling method.

RESULTSThe prevalence of TS among school-age children was estimated to be 0.43% (0.74% for males and 0.07% for females). The prevalence of male children and juveniles was higher than that of female children and juveniles (chi(2) = 25.09, P < 0.001, prevalence ratio = 10.95, prevalence ratio 95% CI: 3.38 - 35.46). The highest prevalence of TS was between 9 - 10 years old. The mean age at onset of TS was 7.7 +/- 2.7 years, with 45.2% of them among 6 - 7 year olds. The rate of delayed diagnosis and rates of misdiagnosis and misclassification of the syndromes were 78.6%, 42.9% and 23.8%, respectively.

CONCLUSIONTourette syndrome had been a common disease of children and juveniles in Wenzhou area. The disease was correlated with age and sex, often misdiagnosed and misclassified. Physicians and as well as general publics should be trained to identify the cases.

Adolescent ; Age Factors ; Child ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Prevalence ; Sex Factors ; Tourette Syndrome ; diagnosis ; epidemiology

Antiviral Agents ; adverse effects ; therapeutic use ; Female ; Hepatitis C, Chronic ; complications ; drug therapy ; psychology ; Humans ; Interferon-alpha ; adverse effects ; therapeutic use ; Mental Disorders ; complications ; Middle Aged

OBJECTIVETo investigate the infection rate and subtypes of human papilloma virus(HPV) in patients with oropharyngeal squamous cell carcinoma (OSCC) and analyze the clinicopathologic features of patients with or without HPV infection.

METHODSA total of 66 biopsy or surgical specimens of OSCC archived in the Pathology Department of Chinese Academy of Medical Sciences were analyzed by polymerase chain reaction (PCR), and the generic amplification products were detected by DNA enzyme immunoassay (DEIA) and typed by reverse hybridization line probe assay.

RESULTSHPV-DNA was detected in 11 (16.7%) of all specimens. Among them, 7 were infected with HPV-16,and the remaining 4 patients were infected with HPV-16/11, HPV-35, HPV-58/52, and HPV-33/52/54, respectively. HPV-16 was detected in 72.7% of all positive specimens. There were more females in HPV-positive group than HPV-negative group (36.4% vs. 1.8%,P=0.002). Patients with HPV-positive tumors were more likely to be non-smokers (36.4% vs. 0,p=0.001) and non-drinkers (45.5% vs. 1.8%,p=0.001) than those with HPV-negative tumors. The proportion of moderately or poorly differentiated tumors was higher in HPV-positive patients than HPV-negative patients (81.8% vs. 63.7%), although without statistical significance (p=0.409). No difference was observed in T classification, N classification, and overall tumor stage.

CONCLUSIONSHPV infection rate was 16.7% in this cohort. HPV-positive OSCC has its unique etiologic and clinicopathological characteristics.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; virology ; DNA, Viral ; isolation & purification ; Female ; Humans ; Male ; Middle Aged ; Oropharyngeal Neoplasms ; virology ; Papillomaviridae ; classification ; isolation & purification ; Papillomavirus Infections ; virology

Objective To explore the relationship between ATP content in CD4+ T lymphocytes and acute rejection after liver transplantation(LT). Methods This study contained 77 patients who received LT from February to October 2009, They were divided into AR (acute rejection) and NAR (non-acute rejection) groups while 56 healthy people were enrolled to serve as the control group.Blood specimens were collected preoperatively and at 1, 2 and 4 weeks postoperatively. For the AR group, specimens were also collected on the day when AR occurred and 1 week after steroid bump together with that of the healthy people. ImmuKnowTM test kits for immune cell function were used to assay the ATP value. Results ATP values within CD4+T lymphocytes were elevated significantly in each group compared with those preoperatively. Peak level was reached in the AR group and was significantly higher than that of the contemporary NAR group (P＜0.05). ROC curve analysis showed that the obvious elevation of the ATP value within CD4+ T lymphocytes 1 week postoperatively had better sensitivity and specificity in diagnosing AR. The ATP sensitivity rate for early AR was 84.6 %and specificity rate 81 %. The ATP value within CD4+ T lymphocytes on the day of AR occurrence had a positive relationship with the rejection acting index(RAI), while relative index (r) was 0. 876(P＜0.05). After the steroid dump treatment, AR in all the patients was reversed and the ATP value declined significantly as compared with the control group and the day when AR occurred(P＜0. 05).Conclusion During the postoperative period, the dynamic change of ATP value within CD4 + T lymphocyte had a close relationship with acute rejection after liver transplantation. Thus, it might be used as a feasible and noninvasive monitoring index for diagnosing AR and the effectiveness of the anti-rejection treatment.