Microbial lipases have been widely used in traditional industries,as well as in emerging biocatalysed areas owing to their ability to catalyze a variety of reactions in aqueous and non-aqueous media.Therefore,it is very important to enhance amount of lipase production by recombinant overexpression for meeting market demand.A critical review of main and novel strategies which have been employed for recombinant expression of microbial lipases are presented,including codon optimization,fusion and co-expression,dual expression system based on hybrid promoters,homologous overexpression,cell surface displaying and high-throughput screening based on gene library of expression.These new technologies are gradually coming to the forefront in the recombinant expression of lipase,especially for cell surface displaying and high-throughput screening based on gene library of expression.Meanwhile,several recombinant expressions for representative microbial lipases were also introduced and discussed,which are available for consultation when attempting to overexpress any lipases by scientists and industrialists.

Objective:To develop an HPLC method for the determination of capsaicine, dihydrocapsaicin, imperatorin and isoim-peratorin in Guanjie Jietong plasters. Methods:A CAPCELL PAK C18 column was used as the chromatographic column, and the flow rate was 0. 8 ml·min-1. The mobile phase A consisted of methanol-acetonitrile(1∶1), the mobile phase B was of 1% phosphoric acid. The detection wavelength was 280 nm and 254 nm, respectively. Results:There was a good linear relationship between the con-centrations and the peak areas within the range of 0. 298-5. 956μg(r=0. 9998) for capsaicine, 0. 152-3. 044μg(r=0. 999 6) for di-hydrocapsaicin, 0. 018-0. 352 μg(r=0. 999 5) for imperatorin and 0. 010-0. 204 μg(r=0. 999 3) for isoimperatorin. The average re-covery was 97. 8%(RSD=1. 02%), 97. 0%(RSD=0. 76%), 96. 6%(RSD=0. 65%) and 98. 1%(RSD=1. 35%), respectively. Conclusion:The method is convenient, accurate, sensitive and repeatable, which can be used in the determination of capsaicine, di-hydrocapsaicin, imperatorin and isoimperatorin in Guanjie Jietong plasters.

Purpose:To study the effect of inhibition of telomerase activity and cell cycle by transcriptase telomerase inhibitors (3′ azido 3′ deoxythymidine, AZT) on squamous cell carcinoma of tongue in vitro.Methods:Human squmous cell carcinoma of tongue cell line Tca8113 was used as target cell. Telomerase activity was determined by TRAP PCR ELISA in untreated and treated Tca8113 by AZT, cell cycle phases were analyzed by flow cytometry. Results:Telomerase activity of Tca8113 was significantly inhibited when treated with AZT, and the effect of inhibition was dose dependant (rate of telomerase activity treated with AZT in 0.3, 0.6, 1.0, 1.5mol 10 -1 was 0.69 0.03, 0.61 0.08, 0.53 0.11, 0.50 0.02 respectively, rate of telomerase activity treated without AZT was 0.76 0.06). Cell cycle of treated Tca8113 was changed with marked increase in G 2 /M phase compared with untreated Tca8113 (62.8% vs 19.7%, P

Objective To investigate the transactivating effect of hepatitis C virus(HCV)core protein on inducible nitric oxide synthase(iNOS)gene promoter and the molecular biological mecha- nisms of HCV pathogenesis.Methods Polymerase chain reaction(PCR)technique was employed to amplify the sequence of iNOS promoter by using HepG2 genomic DNA as template,and the product was cloned into pGEM-T vector.The iNOSp gene was cut from T-iNOSp by KpnⅠand XhoⅠ,and then was cloned into pCAT3-Basic,the constructed vector was named as pCAT3-iNOSp,pCAT3-iN- OSp was transfected into the LO_2 cell line.LO_2 cell was also cotransfected with pcDNA3.1(-)-core and pCAT3-iNOSp by FuGENE 6 transfection reagents.The LO_2 cells transfected with pCAT3-Basic was used as negative control.The activity of CAT in LO_2 cells was detected by an ELISA kit after 48 hours,which reflected the transactivating function of HCV core protein to iNOS gene promoter.Re- sults The expressive vector pcDNA3.1(-)-core and report vector pCAT3-iNOSp had been construc- ted and confirmed by restriction enzyme digestion and sequencing.The expression of CAT in LO_2 cells transfected with pCAT3-iNOSp and peDNA3,1(-)-core was 11 times as higher as that of pCAT3-bas- ic,and 6 times as higher as that of pCAT3-iNOSp.Conclusion It is suggested that HCV core protein can transactivate iNOS gene promoter.

The fermentation conditions of lipase production by Geotrichum candidum Y162 were optimized. Initially, the most suitable carbon olive oil, nitrogen source soybean flour and NH4Cl, salt BaCl2 and MgCl2 were selected according to single factorial experiments respectively. Based on the result, screening methodology Plackett-Burman design was used to evaluate the effects of twelve factors related to lipase production and three statistically significant factors olive oil, BaCl2 and NH4Cl were selected. The path of steepest ascent was used to approach the optimal region of lipase production subsequently. Then, the optimal combined concentration for maximum enzyme activity were further optimized by response surface methodology and determined as follows: olive oil 2.35%, BaCl2 0.36%,and NH4Cl 4.69%.The optimization of culture conditions of G.candidum Y162 led to a 2.25-fold increase in lipase production relative to initial result 14.16 U/ml, which indicate that single factor in combination with response surface methodology is an effective method for optimization of lipase production conditions by G.candidum Y162.

Adult ; Aged ; Humans ; Middle Aged ; Pneumoconiosis ; diagnostic imaging ; Radiography, Thoracic ; Retrospective Studies

To assess the impact of the operational simulation training system on the cultivation of ophthalmic students.
●METHODS: Four hundred and eighty ophthalmic students in China Medical University from 2009 to 2011 were chosen as objects and divided into two groups at random. The students in the experimental group used operation simulation training system combined with traditional teaching methods, while the students in the control group only used traditional teaching methods. We examined the operation skills and the confidence of all the students half a year later.
●RESULTS: The improvement of the operation skills during the suture technique examination, and the confidence for operation technology were significantly different in two groups. The basic skills of the students in the examination groups improved apparently during the operational simulation training and they were full of confidence to their operational technique.
●CONCLUSlON: The operational simulation training is useful for promoting the clinical ability for ophthalmic students and should be widely popularized.

Objective To explore the relationship between peripheral Th17 cell number and chronic gastritis in Helicobacter pylori(H.pylori)-infected children.Methods Children were diagnosed as chronic gastritis by endoscopy.The degree and activity of inflammation were graded by histopathology examinations.The patients with both 13C urea breath test and urease test positive were diagnosed as H.pylori infection.The peripheral Th17 cell number was measured by flow cytometry and expressed as a ratio to total T cell.Results The Th17 cell number in HP group (chronic gastritis with H.pylori infection,n =33),non-HP group (chronic gastritis without H.pylori infection,n =24) and normal controls (n =15) were (1.55 ±0.30)％,(1.06 ±0.33)％,and (1.04 ±0.35)％,respectively.HP group included a statistically higher Th17 cell number than the other groups (all P ＜ 0.05),while no obvious difference was found between non-HP group and controls (P ＞ 0.05).According to the degree of inflammation,the chronic gastritis with H.pylori infection was categorized into non-apparent (n =10),mild (n =8),moderate (n =9) and severe (n =6) subgroups.The Th17 cell number in each subgroup was (1.64 ± 0.21)％ (non-apparent),(1.61 ± 0.23)％(mild),(1.25 ± 0.29) ％ (moderate) and (1.75 ± 0.20) ％ (severe),respectively.The moderate group had a lowest Th17 cell number among 4 groups (P ＜ 0.05).And significant differences did not exit in the other 3 groups (P ＞ 0.05).The HP group patients with different inflammatory activity had a Th17 cell number of (1.23 ±0.25)％ in nonapparent (n=15),(1.53 ±0.15)％ in mild (n=6),(1.55 ±0.32)％ in moderate (n=6) and (1.71 ±0.35)％ in severe (n =6) subgroup,respectively.However,there were no significant differences among 4 subgroups (P ＞ 0.05).Conclusions In the progress of chronic gastritis with H.pylori infection,Th17 cells may play a role as a double-edged sword by protecting and fighting against H.pylori infection and immunopathologic insults.This would provide more insights into the treatment of H.pylori infection.

Objective To assess the diagnostic value of anti-mutated citrullinated vimentin (MCV)antibody for rheumatoid arthritis (RA).Methods Meta analysis.The databases,including PubMed,The Excerpta Medica Database (Embase) and China National Knowledge Infrastructure (CNKI),from January 1990 to January 2013,were employed to search for the studies related to diagnostic value of anti-MCV for RA.The software Meta-Disc1.4 was used to perform Meta-analysis,investigate the sources of heterogeneity and draw the summary receiver operating characteristic (SROC) curve.The Begg's test was applied to evaluate the publication bias by use of Stata1 1 software.Results A total of 27 studies were included.Pooled sensitivity was 0.74[95％ confidence interval (CI) 0.73 to 0.76] and pooled specificity was 0.83 [95％ CI (0.82 to 0.85)].The area under the summary receiver operating characteristic curves was 0.896.Twenty-three literatures which detected both anti-MCV antibody and anti-cyclic citrullinated peptide(antikCCP) antibody show that:the area under curre of anti-MCV antibody and anti-CCP antibody were 0.887 and 0.924 respectively,but there was no significant difference (Z =1.525,P =0.064).Conclusions Results of the meta-analysis demonstrated that the MCV antibody may be a useful parameter in diagnosing RA as it shows high sensitivity and specificity.But there was no statistical difference between the results of anti-MCV antibody and anti-CCP antibody for the RA diagnosis.

Objective To investigate the value of procalcitonin (PCT) on predicting the severity and prognosis in patients with early acute respiratory distress syndrome (ARDS).Methods A prospective observation study was conducted. A total of 113 patients with ARDS undergoing mechanical ventilation admitted to intensive care unit (ICU) of Affiliated People's Hospital ofJiangsu University from October 2012 to April 2016 were enrolled. Based on oxygenation index (PaO2/FiO2), the patients were classified into mild, moderate, and severe groups according to Berlin Definition. Twenty-five healthy volunteers were served as controls. Demographics, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, and Murray lung injury score were recorded. Within 24 hours after diagnosis of ARDS, the serum levels of PCT and C-reactive protein (CRP) were determined by enzyme-linked fluorescence analysis (ELFA) and immune turbidimetric method, respectively. The patients were also divided into survival and non-survival groups according to clinical outcome within 28-day follow-up, and the clinical data were compared between the two groups. Spearman rank correlationwas applied to determine the correlation between variables. The predictive value of the parameters on 28-day mortality was evaluated with receiver operating characteristic curve (ROC). Kaplan-Meier survival curve analysis was used to compare different PCT levels of patients with 28-day cumulative survival rate. Results After excluding patients who did not meet the inclusion criteria and loss to follow-up, the final 89 patients were enrolled in the analysis. Among 89 ARDS patients analyzed, 27 of them were mild, 34 moderate, and 28 severe ARDS. No significant differences were found in age and gender between ARDS and healthy control groups. Infection and trauma were the most common etiology of ARDS (55.1% and 34.8%, respectively). Compared with healthy control group, both CRP and PCT in serum of ARDS group were higher [CRP (mg/L): 146.32 (111.31, 168.49) vs. 6.08 (4.47, 7.89), PCT (μg/L): 3.46 (1.98, 5.56) vs. 0.02 (0.01, 0.04), bothP < 0.01], and the two showed sustained upward trends with the ARDS course of disease. Compared with mild group, severe group had significantly higher APACHE Ⅱ and Murray scores. Spearman rank correlation analysis showed that both serum PCT and CRP in patients with ARDS was correlated well with APACHE Ⅱ score (r values were 0.669 and 0.601, respectively, bothP < 0.001), while PCT was weakly but positively correlated with Murray score (r = 0.294,P = 0.005), but not the case of CRP (r = 0.203,P = 0.052). APACHE Ⅱ score and serum PCT in non-survival group (n = 38) were significantly higher than those of the survival group [n = 51; APACHE Ⅱ score: 26.00 (23.00, 28.50) vs. 21.00 (17.00, 25.00), PCT (μg/L): 6.38 (2.82, 9.49) vs. 3.09 (1.08, 3.57), both P < 0.01], but Murray score and CRP level were similar between survivors and non-survivors. The areas under ROC curve (AUC) of APACHE Ⅱ score and PCT for predicting 28-day mortality were 0.781 and 0.793, respectively, which were better than those of AUC of Murray score and CRP (0.606 and 0.561, respectively, allP < 0.05). The AUC of APACHE Ⅱ score combined with PCT was significantly higher than that of PCT (0.859 vs. 0.793,P = 0.048) or APACHE Ⅱ score alone (0.859 vs. 0.781,P = 0.038). Using a PCT cut-off value of > 4.35μg/L for predicting 28-day mortality, the sensitivity and specificity was 92.2% and 63.2%, respectively, and the positive and negative likelihood ratios were 2.50 and 0.12 respectively. Kaplan-Meier survival curve analysis indicated that the patients whose PCT more than 4.35μg/L, had lower 28-day cummulative survival rate as compared with those with PCT ≤ 4.35μg/L (log-rank test: χ2 = 5.013,P = 0.025).Conclusion The elevated serum PCT level in patients with ARDS seems to be correlated well with the severity of lung injury, and appears to be a useful prognostic marker of outcome in the early phases of ARDS.