Adult ; Aged ; Aged, 80 and over ; Brain Injuries ; surgery ; Dilatation ; methods ; Female ; Humans ; Intensive Care Units ; Male ; Middle Aged ; Tracheostomy ; methods

Objective To investigate functional role and clinical significance of the methylation in the promoter of TPAP gene in the development and progression of pancreatic cancer.Methods Surgically resected specimen from 68 patients who were pathologically diagnosed as pancreatic cancer in Changhai Hospital from July 2006 to August 2009 were collected.The methylation in the promoter of TPAP gene in tumor and nontumor adjacent tissue was detected by methylation specific PCR.Results The methylation rate of tumor and non-tumor adjacent tissue was (0.214 ± 0.057) ％ and (0.084 ± 0.096) ％,respectively,and pancreatic cancer tissue had significantly higher methylation rate than the adjacent tissue.Hypermethylation of TPAP gene was not correlated with age,gender,tumor differentiation,lymphatic metastasis,serum CEA and CA19-9,but was positively correlated with distant metastasis.Conclusions Hypermethylation in the promoter of TPAP gene may participate in the invasion and metastasis of pancreatic cancer and the hypermethylation of the promoter is closely associated with the tumorigenesis and development of pancreatic cancer.

An on-line solid phase extraction ( SPE ) coupled with HPLC-MS/MS method was developed to determine S-ammuxetine and R-ammuxetine in rat plasma. The sample preparation consisted of the following steps:A protein precipitation extraction used methanol and acetonitrile ( 50:50 , V/V ); an on-line SPE treatment to remove most matrixes in plasma;an enrichment and separation step used a C18 analytical column. S-and R-ammuxetine were determined by tandem mass spectrometry. The SPE column was a Retain PEP Javelin (10 mm × 2. 1 mm × 5 μm), while the chromatographic separation was achieved using a ZORBAX SB-C18 (50 mm × 2. 1 mm × 3. 5 μm) analytical column with an isocratic mobile phase composed of acetonitrile-water-formic acid (40:60:0. 1, V/V/V, 0. 3 mL/min). The selected reaction monitoring mode of the positive ion was performed and the precursor to the product ion transitions of m/z 292 . 1/154 . 0 and m/z 260. 4/116. 2 were used to measure S-ammuxetine, R-ammuxetine and internal standard (propranolol). The method was linear over a concentration range from 0 . 2 to 1000 μg/L with the correlation coefficients of 0 . 9903 and 0 . 9951 . The average intra-day precision values were 1 . 2% -12 . 0% for S-ammuxetine and 0. 4%-11. 2% for R-ammuxetine, respectively. The average recoveries were 94. 2%-101. 6% for S-ammuxetine and 94. 3% -109. 4% for R-ammuxetine. Compared to the literature, the sensitivity of this method increased dramatically. The present method has been successfully applied to the preclinical rat research of ammuxetine isomers following intragastric administration.

To establish a stable mouse model of systemic Candida infection and to set up related standard operation procedure .Methods ICR mice were infected with C.albicans or C.parapsilosis by tail vein injection after immunosuppres-sion by cyclophosphamide .The quality control key points included immunosuppression , strain preparation, inoculation doses and the route of inoculation .Survival analysis , bacterial loads and pathological examination were performed to evaluate the prepared model .Results The developed model showed fugal-specific lesions in multiple organs , especially in the kidneys revealed by histopathological examination .Conclusions A stable mouse model of systemic Candida albicans infection can be successfully established by following standardized operation procedure .This mouse model may provide a useful tool for studies on pathogenesis and immune defense of fungal infection and new anti-fungal drug development and so on .

Objective To investigate the incidence of postoperative intubation-related complications and the need for measurement and adjustment of cuff-pressure of endotracheal tube. Methods Five hundred and nine patients of either sex undergoing elective surgery under general anesthesia with tracheal intubation were divided into 2 groups: control group (group C)and cuff-pressure measured group (group CPM). Operation time and duration of endotracheal tube were recorded. Twenty patients with the duration of endotracheal tube 120-180 min in each group were selected. Their tracheal mucous membrane was examined by fiberoptic bronchoscopy at the time of withdrawal of endotracheal tube. Postoperative complications including cough and bloody sputum were also recorded at 24 h after extubation. Results The two groups were comparable with respect to age, sex, operation time and the duration of endotracheal tube. The incidence of sore throat and bloody sputum was significantly higher in group C than in CPM group. The incidence of sore throat and bloody sputum was significantly increased as the duration of endotracheal tube was prolonged in control group while in CPM group only the incidence of sore throat was increased. Varying degrees of injuries to tracheal mucous membrane were observed with fiberoptic bronchoscope.The injuries were more serious in group C than in CPM group. Conclusion Measurement of intracuff pressure is helpful for reducing the tracheal intubation-related complications.

Objective Establishing the drug?resistant Candida albicans disseminative infected mice model for new drug screening. Methods The disseminative infected mouse model was generated by intravenously injecting a clinical Drug?resistant Candida albicans strain ( CaR) to immunosuppressive ICR mice. The features of model was evaluated by clinical symptom, survival condition, fungal burden in tissue, histopathology, cytokines assay and medication. Results After infected with CaR (0 day), the death of mice started at the first day, though, compared to clinical drug sensitive strain ( CaS) infected group, the difference of mortality rate in 16?day observation period was not significant in two groups (CaR, 90?7%;CaS, 86?2%, P =0?158), mice in CaR group died faster than those in CaS group at the early stage;On the fourth day of infection, Candida albicans could be detected in the different tissues, and we found fungal burden in kidney and brain was a significant difference. The typical granuloma caused by fungal infection was the main histopathological feature observed in the kidney, brain and heart. Cytokines in renal tissue were detected by flow cytometry, The changes of IL?1α,IL?6,TNF?αand IFN?γin kidney were significant. Compared with CaS group, IL?1 and IFN?γ were significantly higher and TNF?αdecreased significantly in CaR group. The mice of groups CaR and CaS were treated with 10 mg/kg fluconazole, the mortality rates were 83?3% and 37?5%, which have a significant difference. Conclusions In this study, we successfully established a drug?resistant Candida albicans disseminative infected mice model which is potential tool for the development of new anti?infectious agent.

ObjectiveTo elucidate the association of immune response to hepatitis B vaccination with HLA-DRB1 * 12 allele as well as the level of IL-4 and IFN-γ.MethodsSeventy-four healthy college students from Guangxi province who had non- or hypor -response to recombinant hepatitis B vaccination and 64 medium- or hyper-responders with the conditions of similar were selected randomly and involved in this study.HLA-DRB1 * 12 was detected by PCR-SSP,the level of IFN-γ and IL-4 cytokines were examined by ELISA.Results(1)The allelic frequencies of HLA-DRB1 * 12 was lower in the non- or hypor-responders than that in the medium- or hyper-responders ( 10.8％ vs 32.8％,P=0.002) ; (2)The expression level of IFN-γ in the non- or hypor-responders ( 7.21±7.92 ) ng/ml was much less than that of the medium- or hyper- responders ( 16.36± 11.00) ng/ml ( P=0.000).(3) The expression level of IL-4 in the non- or hyporresponders (3.18±4.45) ng/ml was much less than that of the medium- or hyper- responders (7.76±5.71 ) ng/ml(P=0.000).(4)No significant differences was seen between the expression level of IFN-γ in the HLA-DRB1 * 12 positive ( 13.18± 11.24) ng/ml and the negative ( 11.00± 10.29 ) ng/ml ( P =0.349 ).(5)No significant differences was seen between the expression level of IL-4 in the HLA-DRB1 * 12 positive (5.947±4.530) ng/ml and the negative (5.132±5.800) ng/ml (P=0.423).ConclusionHLA-DRB1 * 12 might be the allele enhanced immune response to hepatitis B vaccination.The expression levels of IFN-γand IL-4 correlating to Thl/Th2 cells might affect on the immune response to hepatitis B vaccination.

OBJECTIVE: To investigate the adenovirus-mediated LacZ gene expression and the destination in different organs of SD rats after the intravenous injection in rats. METHODS: Recombinant adenovirus vector containing LacZ was transferred to SD rats by injecting into the internal jugular vein. To identify the sites and periods of LacZ gene expression, X-gal staining was used to detect beta-gal level and period of LacZ gene expression of different organs in the transfected and non-transfected rats at different time intervals. RESULTS: On the 1st day after the injection, the lung, liver, kidney, and spleen expressed some beta-gal; on the 3rd day after the injection, the lung, liver, kidney, and spleen expressed beta-gal obviously; their peak levels were on the 7th day; the beta-gal level decreased on the 14th day; beta-gal expression disappeared in the most organs except the lungs on the 28th day. In all animals, the brain did not express any beta-gal. CONCLUSION The adenovirus-mediated exogenous gene transfer in the internal jugular vein may be an effective approach of gene therapy in some diseases in the lung, liver, and kidney.
Adenoviridae ; genetics ; Animals ; Female ; Gene Transfer Techniques ; Genetic Therapy ; Injections, Intravenous ; Jugular Veins ; Lac Operon ; genetics ; Lung ; metabolism ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; genetics ; Transfection

Study on features of acupoints with resistance test in the past half century is reviewed in this article. Mechanism and technology of the method are introduced as well as its shortcomings. The determination method of signal transmission along meridians with the combination of electrical network theories and practice is advanced. And the result of a series experiments on one meridian at the superficial part of the body are given as well. Thus, it is concluded that the signals of the point-in/point-out and the signals along a non-meridian path with the same distance are significantly different, which gives a verification of the feasibility of the method by using electrical network theories to set out characteristics of signal transmission along meridians dynamically.
Acupuncture Points ; Electrophysiological Phenomena ; Electrophysiology ; instrumentation ; methods ; Humans ; Meridians ; Signal Transduction

Leishmaniasis is a prevalent cause of death and animal morbidity in underdeveloped countries of endemic area. However, there is few vaccine and effective drugs. Antimicrobial peptides are involved in the innate immune response in many organisms and are being developed as novel drugs against parasitic infections. In the present study, we synthesized a 5-amino acid peptide REDLK, which mutated the C-terminus of Pseudomonas exotoxin, to identify its effect on the Leishmania tarentolae. Promastigotes were incubated with different concentration of REDLK peptide, and the viability of parasite was assessed using MTT and Trypan blue dye. Morphologic damage of Leishmania was analyzed by light and electron microscopy. Cellular apoptosis was observed using the annexin V-FITC/PI apoptosis detection kit, mitochondrial membrane potential assay kit and flow cytometry. Our results showed that Leishmania tarentolae was susceptible to REDLK in a dose-dependent manner, disrupt the surface membrane integrity and caused parasite apoptosis. In our study, we demonstrated the leishmanicidal activity of an antimicrobial peptide REDLK from Pseudomonas aeruginosa against Leishmania tarentolae in vitro and present a foundation for further research of anti-leishmanial drugs.