Objective To investigate the surgical indicatin and outcome of recurrent gastric carcinoma.Methods Review the clinical data of 56 cases with recurrent gastric carcinoma underwent reoperation from 1990 to 2000.The operative indication, surgical method and complications were analized.Results In 25 cases the recurrent tumor were resected (44.6%),in 10 cases adjacent organs were resected.NO operative motality was observed.Conclusions More aggressive radical resection of the gastric cancer is the key to prevent recurrence.Laparotomy should be performed without delay once there is a suspicion of recurrence.

Objective:To explore the effects of omeprazole combined with somatostatin on intra-abdominal pressure and intestinal mucosal barrier function of elderly patients with severe acute pancreatitis.Methods:Selected 82 cases of elderly patients with severe acute pancreatitis from our hospital.They were divided into research group (48 cases) and control group (34 cases).The control group was treated with basic therapy,and the research group was treated with omeprazole combined with somatostatin on the basis of the control group.Analyzed and compared the clinical effects,intestinal mucosal barrier function and symptom indexes of patients in the two groups.Results:①The total effective rate of the research group was significantly better than that of the control group,and the difference was statistically significant (P=0.005).②After treatment,the D-lactose and diamine oxidase (DAO) levels of both groups were significantly lower than before treatment,and the difference was statistically significant (both P＜0.001).In addition,after treatment,the D-lactose and DAO levels were significantly lower in the research group than in the control group,and the differences were statistically significant (mean P ＜ 0.001).③After treatment,patients of the research group showed better results in abdominal pain and abdominal distension relief time,bowel recovery time,hospital stay and blood amylase index than those in the control group,and the difference was statistically significant (mean P ＜ 0.001).④After treatment,the intra-abdominal hypertension of two groups was significantly lower than before treatment,with statistical difference (P ＜ 0.001).Yet it was significantly lower in the research group than in the control group,with statistical difference (P ＜ 0.001).Conclusion:The combined treatment of omeprazole and somatostatin can effectively improve the clinical efficacy of severe acute pancreatitis in elderly patients,correct intra-abdominal hypertension and improve the intestinal mucosal barrier function of patients,so it is worthy of reference and promotion.

Objective To construct HaCaT cell lines stably expressing the wild type human GJB6 gene or its mutant by using a Tet-On lentiviral vector, and to lay an experimental foundation for studies on pathogenesis of hidrotic ectodermal dysplasia. Methods The wild-type human GJB6 gene and its mutant (A88V)were amplified by PCR, and then inserted into the Tet-on lentivirus plasmid to construct recombinant lentivirus vectors. The recombinants were identified by gene sequencing and enzymatic digestion. Cultured HaCaT cells were classified into three groups to be transfected with a negative control lentiviral vector (NC group), the lentivirus vector expressing the wild-type human GJB6 gene (WT group), or the lentivirus vector expressing the mutant human GJB6 gene (MU group). Puromycin was used to select HaCaT cell clones stably expressing the GJB6 gene which encodes the connexin 30 (Cx30)protein. The selected HaCaT cell clones were cultured with or without tetracycline for 48 hours, thereafter, real-time PCR(RT-PCR) was performed to detect GJB6 gene mRNA expression, Western-blot analysis to measure expressions of Cx30 and FLAG-tag proteins, and cell counting kit 8 (CCK8)assay to evaluate cellular proliferative activity. Results Enzymatic digestion and gene sequencing showed that recombinant lentivirus plasmids were successfully constructed. RT-PCR showed evidently increased mRNA expression of the GJB6 gene in stably transfected HaCaT cells. Moreover, the expression abundance of the GJB6 gene was 112.369 times higher in the WT group induced by tetracycline than in that without tetracycline treatment (P < 0.05), and 2.249 times higher in the MU group induced by tetracycline than in that without tetracycline treatment (P < 0.05). Western-blot analysis showed that Cx30 and FLAG-tag proteins were stably expressed in the WT group and MU group after induction with tetracycline, while neither of them was observed in the WT group or MU group without tetracycline treatment, or in the NC group. Significant differences were noted in cellular proliferative activity (expressed as the absorbance value at 450 nm)between the MU group with and without tetracycline treatment and between the WT group with and without tetracycline treatment at 4, 8, 12, 24, 36 and 48 hours (all P <0.05), but not between the NC group with and without tetracycline treatment at any of the above time points (all P >0.05). Conclusion HaCaT cell lines which stably express the wild-type GJB6 gene or its mutant(A88V)are successfully constructed.

Purpose To explore the association of NET-1 expression with the angiogenesis of cervical squamous cell carcinoma and the related molecular mechanism.Methods Immunohistochemistry MaxVision was used to detect the expression of NET-1,NF-κB p65 and VEGF in 80 cases of cervical squamous cancer tissue and 10 cases of chronic cervicitis tissue.The microvascular density (MVD) was assessed by labeling endothelia with CD34-antibody and counted according to Weinner's standard.The eukaryotic expression plasmid pCMV6-NET-1 was transfected into cervical squamous cancer cells by cationic liposome in order to obtain over expression of NET-1 gene.In order to inhibit the activation of NF-kappa B,cervical squamous carcinoma cells were treated with NF-kappa B specific inhibitor PDTC.The protein expression in SiHa cells was assessed by Western blotting.Results The expression level of NET-1 in the cervical squamous carcinoma tissue was significantly higher than that in the tissue of control group.NET-1 expression was positively correlated to the MVD,NF-κB p65 and VEGF expression,respectively.Transient transfection of NET-1 gene significantly increased the nuclear expression of NF-κB p65 and cytoplasm expression of VEGF in the SiHa cells.However,pretreatment of SiHa cells with NF-inhibitor PDTC significantly inhibited NET-1 induced upregulation of VEGF expression.Conclusion NET-1 promotes the angiogenesis of cervical squamous carcinoma through the activation of the NF-κB signaling which upregulate the expression of VEGF in the cancer cells.

Rhamnolipid biosurfactant is mainly produced by Pseudomonas aeruginosa that is the opportunistic pathogenic strain and not suitable for future industrial development. In order to develop a relatively safe microbial strain for the production of rhamnolipid biosurfactant, we constructed engineered Escherichia coli strains for rhamnolipid production by expressing different copy numbers of rhamnosyltransferase (rhlAB) gene with the constitutive synthetic promoters of different strengths in E. coli ATCC 8739. We further studied the combinatorial regulation of rhlAB gene and rhaBDAC gene cluster for dTDP-1-rhamnose biosynthesis with different synthetic promoters, and obtained the best engineered strain-E. coli TIB-RAB226. Through the optimization of culture temperature, the titer of rhamnolipd reached 124.3 mg/L, 1.17 fold higher than that under the original condition. Fed-batch fermentation further improved the production of rhamnolipid and the titer reached the highest 209.2 mg/L within 12 h. High performance liquid chromatography-mass spectrometry (LC-MS) analysis showed that there are total 5 mono-rhamnolipid congeners with different nuclear mass ratio and relative abundance. This study laid foundation for heterologous biosynthesis of rhanomilipd.
Bacterial Proteins ; genetics ; Batch Cell Culture Techniques ; Decanoates ; Escherichia coli ; metabolism ; Fermentation ; Glycolipids ; biosynthesis ; Hexosyltransferases ; genetics ; Industrial Microbiology ; methods ; Multigene Family ; Promoter Regions, Genetic ; Pseudomonas aeruginosa ; Rhamnose ; analogs & derivatives ; biosynthesis ; Surface-Active Agents ; metabolism

Objective To investigate the impact of neoadjuvant chemotherapy on surgery and prognosis in patients with stage Ⅲ gastric cancer.Methods The 90 cases staging Ⅲ gastric cancer were randomly divided into neoadjuvant chemotherapy group and surgery group.Both surgical complications and prognosis were compared.Results The overall effective rate of neoadjuvant chemotherapy was 53％ (24/45),disease control rate of neoadjuvant chemotherapy was 91％ (41/45).Blood loss (t =4.102,P =0.037),local adhesions surgery (x2 =19.756,P =0.000),local tissue necrosis (x2 =13.512,P =0.000),tissue fragility (x2 =12.870,P =0.000) number of cases found in neoadjuvant chemotherapy group were higher than surgery group,and the data were statistically significant.There were no statistically significance between two groups about operation time (t =2.391,P =0.129),tissue congestion (x2 =0.865,P =0.352),postoperative bleeding(x2 =0.720,P =0.396),postoperative fistula (x2 =1.047,P =0.306).The survival time of neoadjuvant chemotherapy group was longer than surgery group,but it was not statistically significant(t =1.086,P =0.372).Conclusions Neoadjuvant chemotherapy could reduce the stage of the gastric cancer and increase the complexity of surgery.Thus preoperative evaluation should be prepared before the surgery.

Objective To establish a method for the determination of protocatechuic acid, salidroside, and chlorogenic acid in Sargentodoxa cuneata. Methods The separation was performed on a Waters XSELECT CSH C18 (150 mm × 4.6 mm, 5 μm) with methanol-acetonitrile-0.2 % phosphoric acid as the mobile phase in a gradient elution at a flow rate of 0.8 ml/min. The detection wavelength was 260 nm and the column temperature was 35 ℃. Results The linear ranges of protocatechuic acid, salidroside, and chlorogenic acid were 0.0020-0.0120, 0.0600-0.3602, 0.0750-4.5006 mg/ml, respectively. The average recoveries were 98.01% (RSD=0.07%), 98.53 % (RSD=0.12%), and 101.10 % (RSD=1.92%), respectively. Conclusions The method is simple, accurate, and highly reproducible, which could provide the scientific evidence for the quality control of Sargentodoxa cuneata.

A sensitive and selective method was developed for the separation and characterization of related substances (RSs) in EVT-401 by hyphenated LC–MS techniques. Complete separation of the RSs was achieved with an Inertsil ODS-SP column (250 mm×4.6 mm, 5 μm) by linear gradient elution using a mobile phase consisting of 0.2% formic acid solution, methanol and acetonitrile. EVT-401 was found to be susceptible to acid, alkaline and oxidative stresses, while relatively stable under photolytic and thermal dry stress conditions. Fourteen RSs including six process-related substances and eight degradation products were detected and identified in EVT-401 with positive ESI high-resolution TOF-MS analysis of their parent ions and the corresponding product mass spectra elucidation, and some of them were further verified by chemical synthesis and NMR spectroscopy. The specific LC–MS method developed for separation, identification and characterization of RSs is valuable for EVT-401 manufacturing process optimization and quality control.

This study aimed to identify the related substances of lorazepam tablets by liquid chromatography mass spectrometry (LC-MS). To separate the related substances of lorazepam tablets, gradient elution was performed using acetonitrile and 0.1% acetic acid -20 mmol/L of ammonium acetate as mobile phase on Inert Sustain C18 (250 mm × 4.6 mm, 5 μm).The accurate mass and elemental composition of the parent ions and their product ions of related substances were determined by electrospray-ionization quadrupole time-of-flight high resolution mass spectrometry (ESI-Q-TOF/MS).The structures of the related substances were identified by spectral analysis. Under the established analytical condition, lorazepam and its related substances were adequately separated, and 22 major related substances with content greater than 0.1% were detected and identified by hyphenated techniques in lorazepam tablets and their stressed samples.Among them, 5 were the impurities listed in the USP or EP, and the others were unknown related substances identified for the first time in this paper.The LC-MS technique can effectively separate and identify the related substances of lorazepam tablets, which provides some reference for quality control.

To establish a method for the determination of formaldehyde and glyoxal simultaneously in varenicline tartrate active pharmaceutical ingredient (API) and its intermediate, formaldehyde and glyoxal were derivatized by 2, 4-dinitrophenylhydrazine (2,4-DNPH) to improve the HPLC retention and UV detection sensitivity. Separation was performed on a C8 (150 mm × 4.6 mm, 5 μm) column by linear gradient elution using acetonitrile and water as the mobile phase; the detective wavelength was set at 380 nm.Formaldehyde and glyoxal were quantitatively determined by an external reference method.Linear calibration was established for both formaldehyde and glyoxal in the range from 0.094 to 1.88 μg/mL.The detection and the quantification limits were 0.047 μg/mL (19 μg/g) and 0.094 μg/mL (38 μg/g), respectively.The recoveries were (95.0±1.1)% and (99.4 ± 2.6)% for formaldehyde and glyoxal, respectively.This method has been fully validated to be applicable to quantitative analysis of trace amount of formaldehyde and glyoxal in varenicline tartrate API and its intermediate.Test results demonstrated that the contents of both formaldehyde and glyoxal met the permitted daily exposure (PDE) limits for the finished products of varenicline tartrate API as well as its intermediate, though the glyoxal contents in the crude intermediates were likely to exceed the limit.The established method is valuable for the manufacturing process and quality control of varenicline tartrate.