Objective To investigate the effects of transforming growth factorβ1(TGFβ1)and β3 (TGFβ3)gene transfer on MMP-2,MMP-9 and TIMP-1 expression in hepatic stellate cells(HSC-T6).Methods TGFβ1 and TGFβ3 expression plagmids were constructed.The recombinant expression plasmid pcDNA3.1(+)-=TGFβ1 and pcDNA3.1(+).TGFβ3 were transfected or cotransfected into HSC-T6.At 24,48 and 72 h after transfection,the expression of MMP-2,MMP-9 and TIMP-1 mRNA were detected by real-time quantitative PCR,and the expression of MMP-2,MMP-9 and TIMP-1 protein were detected by Western blot.The recombinant expression plasmid pcDNA3.1(+).TGFβ1 was transfected into HSC-T6,and positive clones were selected by G418.The positive clones were transfected by the recombinant expression plasmid pcDNA3.1(+).TGFβ1,and the expression of MMP-2,MMP-9 and TIMP-1 were detected at 48 h after transfection.Results After transfection with peDNA3.1-TGFβ1,MMP-2 and TIMP-1 increaged remarkably in HSC-T6 cells(P＜0.05),but MMP-9 remained at the sanle level;After transfection with pcDNA3.1-TGFβ3,expression levels of MMP-2,MMP-9 and TIMP-1 mRNA were not changed,but TIMP-1 protein increased remarkably(P＜0.05);in cotransfection group,the expression of MMP-2 was higher than that in the blank and the control groups(P＜0.05),but MMP-9 level was not changed and TIMP-1was decreased compared with that in the TGF-β1 transfection group(P＜0.05).After TGFβ3was transfected into positive clones,the change of MMP-2 wag not significant(P＞0.05).but MMP-9 increaged and TIMP-1 decreased significantly at 48 h after transfection(P＜0.05).Conclusions TGFB3 may inhibit liver fibrosis by increase the activity of MMP-2 and MMP-9,and decrease the activity of TIMP-1.

Asthma ; prevention & control ; therapy ; Child ; Disease Management ; Family ; Humans

Standardized training of residents is a necessary way for clinical medical personnel train-ing. Residents graduating from medical colleges and universities should accept the standardized, scientific, strict pre-service training, in order to realize role conversion from medical students to hospital physicians. This paper summarizes and analyzes the pre-service training, focusing on the training object, course design, organization implementation, evaluation, questionnaire survey, and puts forward some thoughts and sugges-tions on strengthening and improving pre-service training form, expanding the content of training, and quan-tifying the evaluation of training effect.

Objective To investigate the relationship between extracellular matrix metalloproteinase inducer (EMMPRIN), the upstream regulatory factor of matrix metalloproteinase (MMPs), and the formation of atherosclerosis and the clinical type of coronary heart disease. Methods A total of 223 patients were classified into four groups according to results of coronary angiography (CAG) and clinical data: STEMI group (65 patients with ST-segment elevation myocardial infarction), NSTE ACS group (42 patients with non-ST-segment elevation acute coronary syndrome), SAP group (75 patients with stable angina pectoris) and normal control group (41 patients of CAG-negative). The mean fluorescence intensity (MFI) of EMMPRIN on monocytes of peripheral blood (PBMCs)were examined by flow cytometry. MMP-9 in serum was measured with ELISA; high-sensitivity C-reactive protein (hs-CRP) in serum was measured with immune velocity method. Results The EMMPRIN MFI on PBMCs in SAP group, STEMI group and NSTE ACS group was higher than that in the normal control group (P<0.05 or P<0.01). The EMMPRIN MFI in STEMI group and NSTE ACS group was higher than that in SAP group (P<0.05 or P<0.01). The expression characteristic of EMMPRIN on the PBMCs was consistent with that of hs-CRP and MMP-9 in each group. The EMMPRIN MFI of the PBMCs had positive correlation with the level of MMP-9 and hs-CRP in serum (r=0.168,P<0.05;r=0.305,P<0.01). Conclusion EMMPRIN may has promotive effect on the formation of atherosclerosis and unstablility of coronary heart disease as an upstream regulatory factor of MMPs

Heat-shock protein (Hsp) 70 potentiates specific immune responses to some antigenic peptides fused to it. Here, the prokaryotic plasmids harboring the envelope glycoprotein E0 gene of classical swine fever virus (CSFV) and/or the Hsp70 gene of Haemophilus parasuis were constructed and expressed in Escherichia coli Rosseta 2(R2). The fusion proteins were then purified. Groups of Balb/c mice were immunized with these fusion proteins, respectively, and sera collected 7 days after the third immunization. Immune effects were determined via an enzyme-linked immunosorbent assay and flow cytometric analyses. E0-Hsp70 fusion protein and E0+Hsp70 mixture significantly improved the titer of E-specific antibody, levels of CD4+ and CD8+ T cells, and release of interferon-γ. These findings suggested that Hsp70 can significantly enhance the immune effects of the envelope glycoprotein E0 of CSFV, thereby laying the foundation of further application in pigs.
Animals ; Antibodies, Viral ; blood ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Cell Proliferation ; Classical swine fever virus ; genetics ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Haemophilus parasuis ; genetics ; Immunization ; Interferon-gamma ; metabolism ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics

OBJECTIVE:To determinate the release rate and in vitro transdermal rate of asarinin in Cancer pain cataplasm. METHODS:Using homemade devices and modified France diffusion,isolated skin of rats as barrier,normal saline as solvent,the content of asarinin was determined by HPLC. Release rate of Cancer pain cataplasm within 20,50,80 and 120 min and transder-mal amount within 2,4,8,12,24 h were investigated,and accumulative release rate and accumulative transdermal rate were cal-culated. RESULTS:Accumulative release rate by 120 min of asarinin in Cancer pain cataplasm was 73.01%;24 h in vitro transder-mal rate was 26.01%,and transdermal kinetics equation of asarinin was Q=5.717 7t1/2-0.385 4(r=0.979). CONCLUSIONS:Cancer pain cataplasm has good release and transdermal performance. Its transdermal kinetics is in line with Higuchi equation.

Objective To explore the risk and treatment of laparoscopic cholecystectomy(LC) in patients with liver cirrhosis and cholelithiasis.Methods To summarize the clinical data of 28 patients with liver cirrhosis and cholelithiasis.The patients were repeated right upper quadrant pain,including 4 cases of gallbladder neck stones incarcerated,gallbladder effusion.Among them,there were 20 cases of hepatitis B cirrhosis,5 cases of schistosomiasis cirrhosis,3 cases of alcohol.Results There were 6 patients with hemorrhage during operation.Among them,5 patients were treated with gelatin sponge,hemostatic gauze and bio-glue spray to stop bleeding,1 case was transferred to open surgery because of bleeding.The remaining 27 cases of LC were successful.Complications occurred in 8 patients,3 cases of increased liver function abnormalities,1 case of upper gastrointestinal bleeding,1 case of mild hepatic encephalopathy,2 cases of significant ascites formation,1 patient underwent subtotal resection of the gallbladder with Hartmanns bag,and bile was found in the ascites after surgery,but the bile disappeared after five days.All patients with complications after symptomatic treatment were cured,no death,no major bleeding and liver failure,bile duct injury,severe biliary fistula and other serious complications.Conclusion Although the risk of LC in patients with liver cirrhosis is higher than that in the general population,with the help of current high-tech surgical instruments,we can safely accomplish this with an improved surgical procedure.As these patients require high technical requirements of surgery,surgeons must have a wealth of experience and familiar laparoscopic liver and gallbladder anatomy.

Objective To investigate the efficacy and safety of citrate anticoagulation for continuous renal placement therapy (CRRT) in patients at high risk of bleeding. Methods Forty-seven patients who needed to CRRT and were admitted into the department of critical care medicine from January 2015 to October 2016 were enrolled in this study. According to the patient′s actual condition, they were divided into citrate groups (24 cases) and saline group (23 cases). Patients in saline group were given saline wash. The efficacy and safety were compared between the two groups. The filter lifetime, after treatment the activated partial thromboplastin time (APTT), hemoglobin (Hb), blood gas indexes were compared between the two groups. Results The citrate group used 76 filters while the 0.9% sodium chloride group used 87 filters. The average filter lifetime in citrate group was (22.4 ± 12.7) h which was longer than (8.6±3.3) h in 0.9%sodium chloride group (t=9.79, P<0.01). The incidence of coagulation in extracorporeal circulation was 3.9%(3/76) which was lower than 16.1%(14/87) in 0.9%sodium chloride group(χ2=5.20, P<0.05). Conclusions Regional citrate anticoagulation is a safe and effective option for CRRT in patients at high risk of bleeding.

Objective To observe the effects of mind-regulating brain-developing acupoint magnetic therapy in patients with mild cogni-tive impairment (MCI). Methods From May to October, 2015, 60 patients with MCI were randomly divided into control group (n=30) and observation group (n=30). The control group received magnetic therapy on Yangming meridians, and the observation group received magnet-ic therapy on mind-regulating brain-developing acupoints, for two months. They were assessed with Montreal Cognitive Assessment (Mo-CA), digit-span and digit symbol of Wechsler Adult Intelligence Scale (WAIS). Results After treatment, the scores of MoCA, digit-span and digit symbol of WAIS significantly increased in the observation group (t>4.858, P<0.001), and were higher than that in the control group (t>2.188, P<0.05), however, only the scores of MoCA and digit symbol of WAIS significantly increased in the control group (t>5.527, P<0.001). The scores of visual space and execution, and delayed recall of MoCA increased in the observation group (t>2.324, P<0.05), and were higher than that in the control group (t>2.262, P<0.05) after treatment, and no significant difference was found in other domains (P>0.05). Conclusion Magnetic therapy on acupoints related with cognition could improve the cognitive function in patients with MCI.

Although empirically well understood in their clinical administration, volatile anesthetics are not yet well comprehended in their mechanism studies. A major conundrum emerging from these studies is that there is no validated model to assess the presumed candidate sites of the anesthetics. We undertook this study to test the hypothesis that the single-celled Paramecium could be anesthetized and served as a model organism in the study of anesthetics. We assessed the motion of Paramecium cells with Expert Vision system and the chemoresponse of Paramecium cells with T-maze assays in the presence of four different volatile anesthetics, including isoflurane, sevoflurane, enflurane and ether. Each of those volatiles was dissolved in buffers to give drug concentrations equal to 0.8, 1.0, and 1.2 EC50, respectively, in clinical practice. We could see that after application of volatile anesthetics, the swimming of the Paramecium cells was accelerated and then suppressed, or even stopped eventually, and the index of the chemoresponse of the Paramecium cells (denoted as I ( che )) was decreased. All of the above impacts were found in a concentration-dependent fashion. The biphasic effects of the clinical concentrations of volatile anesthetics on Paramecium simulated the situation of high species in anesthesia, and the inhibition of the chemoresponse also indicated anesthetized. In conclusion, the findings in our studies suggested that the single-celled Paramecium could be anesthetized with clinical concentrations of volatile anesthetics and therefore be utilized as a model organism to study the mechanisms of volatile anesthetics.