Objectives: To observe the roles of nuclear factor ?B(NF ?B) in corticosterone injured hippocampal neurons. Methods:Two groups of primary hippocampal neurons were cultured for 8～10 days. One group was treated with 1?10 -5 mol/L corticosterone. The total cellular nuclear protein levels were extracted at 0.5,1,2 and 4 hours, respectively. Electrophoresis mobility shift assay (EMSA) was used to detect the expression of NF ?B activation. Another group was treated with ?B decoy DNA(25 ?mol/L) to inhibit the activity of NF ?B for 2 hours before 1?10 -5 mol/L corticosterone was intervened. MTT was used to observe the effects of corticosterone induced neurons injury after 6,12,24 and 48 hours. Results:The activity of NF ?B became significantly lower than that in the control group after corticosterone acted on neurons for 1 hour. As time went on, its effects became more conspicuously. The injury of hippocampal neurons aggravated when the activity of NF ?B was inhibited by ?B decoy DNA. Conclusions: NF ?B plays a protective role in corticosterone induced hippocampal neuron injury.

Objective To study the mechanism of GuiJi cream in dispelling effects on corrupt microorganism.Methods A total of 90 full-layer skin injured rats were served as test model. The effects of Guiji cream on collagen type Ⅰ (C Ⅰ), matrix metalloproteinase-1 (MMP-1) and MMP-3 in granulation tissue of wound were observed with immunohistochemistry technique and image-analysis system at various stages of healing. Results The expression of C Ⅰ was obviously increased and the level of MMP-1 and MMP-3 increased at the early stage and kept at a high level from day 3 to day 10. Conclusion The mechanism of GuiJi cream for dispelling effects on corrupt microorganism was increasing the C Ⅰ content in granulation tissue of wound through inhibiting the secretion of MMP- 1.

Objective:To study the effect of artificial gastric juice on the dissolution of schizandrin A to provide the parameters for the best extraction method of Schisandra chinensis. Methods:Schisandra chinensis was respectively extracted by artificial gastric juice and water. Schizandrin A in the extracts was determined by HPLC, and the dissolution of schizandrin A in artificial gastric juice and water was studied and compared. Results:At 60 min, schizandrin A dissolution was 0. 483% in artificial gastric juice, and 0. 362%in water. Conclusion:The dissolution of schizandrin A in artificial gastric juice is 33. 4% higher than that in water, suggesting artifi-cial gastric juice can significantly improve the dissolution of schisandrin A.

Objective: To clone human carboxypeptidase A1 and its active center gene as well as construct recombinant vector for expression before analysis their activities. Methods: CPA1 and CPAlactive center gene were amplified by RT-PCR from pancreas tissue, and then sequencing was carried out. The correct target genes were cloned into prokaryotic vector pGEX-4T-1 and transformed into E. coli BL21 before sequence analysis. After induced by IPTG, gene products were analyzed by SDS-PAGE. Target expressed proteins were denaturated, renaturated, purified and evaluated through MTT and agar colony form test. Results: Human carboxypeptidase Al and its active center gene were cloned successfully. New expected protein band of Mr 66,000 and 46,000 appeared on SDS-PAGE after inducement. Both of the expressed proteins have catalytic activity in vitro, but the activity of the latter is inferior when applied to tumor cells. Conclusions: Human carboxypeptidase A1 and its active center gene were cloned successfully. Their prokaryotic expression products were obtained too. The expressed proteins have catalytic activity in. vitro. A new prosperous beginning of further improvement for CPA1 therapy system has been established based on CPA1 and its active center gene in terms of ADEPT against prostate cancer to clinical application.

Objective To observe the occurrence of cardiovascular adverse events in patients undergoing mechanical ventilation with dexmedetomidine sedation,and to evaluate its safety in intensive care unit (ICU).Methods A prospective randomized controlled trial was conducted.Adult critical patients undergoing mechanical ventilation over 48 hours admitted to ICU of Zigong First People's Hospital in Sichuan Province were enrolled.The patients were divided into dexmedetomidine group (Dex group) and midazolam group (Mid group) according to the randomise number generated by computer.The patients in both groups were given slow intravenous infusion of 0.05 mg/kg midazolam and 1-2 μg/kg fentanyl to induce anesthesia before tracheal intubation,followed by 0.06 mg· kg-1· h-1 midazolam and 20-50 μg· kg-1 · h-1 fentanyl or 0.1-0.2 μg· kg-1 · h-1 sufentanil for continuous intravenous pumping to maintain analgesia and sedation;on the next day,the patients in Dex group was given dexmedetomidine (with the initial dose of 0.4 μg· kg-1 · h-1,and maintenance dose of 0.1-0.7 μg· kg-1 · h-1),and midazolam was stopped half an hour later;the original sedation and analgesia plan remained unchanged in Mid group.The goal of sedation was to maintain a Richmond agitation-sedation scale (RASS) score of-2 to 1 or a Ramsay sedation score of 3 to 4;patients were given midazolam if obvious agitation occurred,in combination with propofol for sedation if necessary;wakeup test was performed every day.Observation endpoints included patients discharged from ICU,death or mechanical ventilation over 28 days.Occurrence of cardiovascular adverse events during sedation such as hypertension,hypotension,bradycardia,tachycardia and arrhythmia,dose of sedatives and analgesics,duration of mechanical ventilation,length of ICU stay and 28-day mortality were observed in two groups.Results A total of 383 patients were enrolled,with 190 patients in Dex group and 193 in Mid group.There was no statistically significant difference in general data such as gender,age,acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and predicted mortality between two groups.Doses of midazolam,propofol and fentanyl in Dex group were reduced as compared with those of Mid group,while the dose of sufentanil was similar in two groups.The Ramsay score and RASS score of Dex group were significantly reduced as compared with those of Mid group (3.34± 0.63 vs.3.95 ± 0.86,-1.33 ±0.87 vs.-1.98 ± 1.27,both P ＜ 0.01).Bradycardia was prominent in Dex group,which was observed in more than half of patients at the beginning of the treatment (1-2 hours),but the patients requiring isoprenaline treatment for heart rate lower than 50 bpm were less than that of Mid group (3.7％ vs.5.2％,x 2 =0.506,P =0.477).The incidences of hypotension (45.3％ vs.68.4％),tachycardia (16.3％ vs.33.7％) and arrhythmia (14.7％ vs.31.1％) in Dex group were significantly lower than those of Mid group (all P ＜ 0.01),and no other cardiovascular adverse events such as cardiac arrest,sinus arrest,hyperglycemia or hypoglycemia were observed in two groups.There was no statistically significant difference in duration of mechanical ventilation between Dex group and Mid group [days:5 (2,28) vs.4 (2,56),Z =-1.917,P =0.055],but compared with Mid group,the length of ICU stay in Dex group was significantly prolonged [days:9 (2,67) vs.6 (2,57),Z =-4.302,P =0.000],and the 28-day mortality in Dex group was significantly reduced (22.6％ vs.44.6％,x2 =20.610,P =0.000).Conclusion Long time dexmedetomidine sedation is safe in critical patients undergoing mechanical ventilation,which can significantly reduce cardiovascular adverse events except bradycardia,and lower the 28-day mortality.

Objective To construct fusion gene and prokaryofie expression plasmid encoding the Rv1884c and Rv0867c genes in Mycobacterium tuberculosis (M.tb).The fusion protein wsg expressed efficienfly in E.coli cells.Methods The Rv1884c and Rv0867c genes were amplified by polymerase chain reactions (PCR) with specific primers from genomic DNA of M.tb H37Rv strain,and cloned into pGEX-4Tland pUC19 vectors.Rv1884c and Rv0867c were subcloned into the expression vector pGEX-4T-1 and pPRO-EXHT followed by DNA sequencing.The plasmids were transformed into E.coli DH5α and induced to produce GST-fused Rv1884c and His-fused Rv0867c fusion protein.The protein molecular weight and expression format was analyzed by SDS-PAGE.Results The recombinant expressive vectors pGEX-4T-1Rv1884c and pPRO-EXHT-Rv0867c were constructed.The DH5α strains of E.coli with recombinant plasmid showed high level of Rv1884c and Rv0867c gene expressions after IPTG induction.The SDS-PAGE showed that the plasmids expressed Rv1884c and Rv0867c fusion proteins with molecule weight of 45 000 and 80 000.The recombinant protein accounted for 18.3%and 23.7%of total bacteria protein.The expressed proteins could be purified via GSTrao FF and Ni2+-NTA system kits in denatured condition.Conclusions Mycobacterium tuberculosis Rv1884e and Rv0867c genes have been cloned and expressed Successfully in E.coli DH5α.The results lay a basis for further study of fast culfivation in Mycobacterium tuberculosis and investigation of their activities and functions.

Objective To investigate the stress distribution in a cemented femoral component in elderly patients with a partial marrow type Ⅰ intertrochanteric fractures and to compare differences in the stress distribution after a long-and short-stem prosthetic replacement.Methods Spiral CT images of right femur of the volunteer were obtained and processed with Mimics software and modeling software to reconstruct a three-dimensional model of the femur.Based on this,a three-dimensional physical model for partial marrow type Ⅰ intertrochanteric fractures,long-and short-stem femoral prostheses,and cement mantle was established.Subsequently,a three-dimensional finite element model of long-and short-stem prosthetic replacement for intertrochanteric fractures was established using the finite element analysis software,and biomechanical analysis was implemented for the model.Results No significant changes in stress distribution of the femur occurred after long-or short-stem prosthetic replacement.Stress on the femur still presented a gradual increase from the proximal end to the distal end,peaked at the lower 1/3 of the medial and lateral junction and then decreased at the end.A stress concentration zone formed in the medial and lateral end of cement-stem interface in short-stem prosthetic replacement.Besides,the maximum value of stress in the lateral interface reached 15.3 MPa,but without surpassing the fatigue strength of the bone cement.Whereas,a stress concentration zone formed in the distal medial and lateral part of cement-stem interface and the medial middle part of the interface in long-stem prosthetic replacement and the maximum value was also lower than the fatigue strength of bone cement.No significant stress concentration zones were found in the femoral calcar reconstructed using bone cement.Conclusions Cemented long-or short-stem prosthetic replacement achieves no significant alteration in stress distribution of the femur.Loosening probability of the cemented long-or short-stem prosthesis is almost the same,but the latter has advantages of shorter surgery time,minor trauma,and fewer complications and may be more suitable for treatment of the elderly patients with a partial marrow type Ⅰ intertrochanteric fractures.

Objective To explore the surgical indication,diagnosis and treatment of suspected pancreatic cystic lesions.Methods To-tally 341 patients were admitted into our hospital because of pancreatic cystic lesions from October 2010 to October 2015.Except the 278 confirmed cases,the clinical data of the rest 63 patients with vague diagnosis were retrospectively analyzed.Results The surgical indication of pancreatic cystic lesions were:the lesion diameter was more than 4 cm;the imaging diagnosis was malignant;the lesion was combined with obvious clinical symptoms which can not explain with other diseases;there were asymptomatic pancreatic cystic lesion and main pancre-atic duct dilation without surgical risk factors;the follow-up showed that the desease is in development.Endoscopic ultrasonography guided fine needle aspiration is a useful supplement for more accurate diagnosis.However,it is an uncertain diagnostic value currently.Conclusion Diameter of lesion,uncertain imaging diagnosis,severe abdominal symptoms,obstructive jaundice and abnormal serum tumor markers are the surgical indication for patients.

ObjectiveTo screen out the two-component system associated with drug resistance of Mycobacterium tuberculosis by detecting the differential expression of two-component system regulator genes between multidrug resistant Mycobacterium tuberculosis strains and drug sensitive strains. MethodsTotal RNA of MTB was extracted from cultured MTB during the logarithmic phase in the 7H9 brook medium, and then its purity was identified. Reverse transcription was further completed. The expressing levels of TCS response regulators were quantified using SYBR Green I qRT-PCR, which aimed at finding the differential expressions between multidrug resistant strains and sensitive strains. Finally, all of differentially expressed TCS were screened out under the stress of INH, SM and LFA. Results Compared with sensitive strains,multidrug resistant strains of Rv0491, Rv3133c, Rv3143 and Rv3246c were up-regulated 1. 03, 7.11,3.48and 1.37 folds, respectively (t/t' =5. 623, -4. 196, -3. 559 and -3. 016, respectively, P ＜0. 01 ). The expressing level of other regulators had no statistical significance between muhidrug resistant strains and drug sensitive strains. Under the antibiotic pressure, the expression of Rv1027c, Rv3246c and Rv3143 showed significant changes compared with no antibiotic group. ConclusionRv3246c and Rv3143 may be associated with MTB drug resistance and the differentially expressed genes in multi-drug resistant strains may be used as potential drug targets against drug resistant tuberculosis.

BACKGROUND:Previous study has confirmed that seasonal variation and surfactant associated protein A(SP-A)and interleukin-6(IL-6)exhibit a significant role in lung immune and defense function.However,the effect mechanism of them remains unclear.OBJECTIVE:By observing the changes of SP-A and IL-6 of male rats in autumn and winter.to explore the effects of seasonal changes on the nonspecific immunity of lung of normal rats,and to provide new ideas for the experimental basis to the cognition of pathology and physiology mechanism for seasonal attack of respiratory system diseases.METHODS:Male SD rats were purchased from each seasons and divided into groups in chronological order All rats were housed with normal forage and drank freely under room temperature.Then the rats were sacrificed by decapitation before 18 o'clock at corresponding solar term and the lung tissues were analyzed by RT-PCR.RESULTS AND CONCLUSION:Compared to the other groups.the expression of SP-A and IL-6 in the Winter Begins group were smaller.which demonstrated that immune function of lung was existed the seasonaI rhythm of lower in winter and higher in autumn.The SP-A and IL-6 are main material basis of immune function.