Objective To approach the effect of ulinastatin (UTI) on protection of vascular endothelial cells in rats with sepsis and its mechanism.Methods Fifty-two Sprague-Dawley (SD) male rats were randomly divided into a normal saline pretreatment group (control group) and a UTI pretreatment group (UTI group), each groupn = 26. The rats in two groups were given lipopolysaccharide (LPS, 10 mg/kg) intra-peritoneal injection for the establishment of rat septic models. In UTI group, 18 hours before LPS injection, intraperitoneal injection of UTI 100 kU/kg dissolved in 5 mL normal saline was given, while in the control group, 3 hours before LPS injection, intraperitoneal injection of 5 mL normal saline was given to the rats for pretreatment. Respectively, at 0.5, 2, 4, 12, 24, 72 hours after model establishment, tail venous blood and myocardial tissue were taken. The levels of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-10), vascular cell adhesion molecule (VCAM) and intercellular adhesion molecule-1 (ICAM-1) were detected by enzyme-linked immunosorbent assay (ELISA); the correlation between TNF-α and ICAM-1 was analyzed; the expression of ICAM-1 in myocardial cell was determined by immunohistochemistry.Results After model establishment, the levels of TNF-α, IL-6, IL-10, ICAM and VCAM in two groups were gradually increased, reaching the peaks at 24, 12, 12, 72, 72 hours, respectively. Compared with control group, the levels of TNF-α, IL-6, ICAM-1, VCAM of UTI group were significantly lower at various time points [24 hours TNF-α (ng/L): 119.8±28.9 vs. 190.2±30.4, 12 hours IL-6 (ng/L): 327.8±26.9 vs. 948.7±63.8, 72 hours VCAM (ng/L): 36.3±3.2 vs. 68.8±2.4, 72 hours ICAM-1 (ng/L): 115.6±11.6 vs. 129.4±8.2,P < 0.05 orP < 0.01], IL-10 was significantly increased [12 hours (ng/L): 80.7±1.9 vs. 42.3±4.9,P < 0.01]. TNF-αwas positively correlated to ICAM significantly (UTI group:r = 0.907,P = 0.050; control group:r = 0.961, P = 0.010). Immunohistochemistry showed that after modeling for 0.5 hour, basically no positive expression of ICAM-1 in myocardial cells was found in the two groups; in the control group, at 12 hours the positive expression of ICAM-1 was increased, and in UTI group, a little expression of ICAM-1 was seen; at 72 hours, the expression of ICAM-1 was significantly increased in both groups.Conclusion UTI can protect the function of endothelial cells in rats with sepsis by regulating the expressions of proinflammatory cytokine, anti-inflammatory cytokine, adhesion molecules, and improving the microvascular permeability.

Objective To evaluate the capability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry ( MALDI-TOF MS) for rapid identification of methicillin-resistant Staphylo-coccus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) strains.Methods Twenty-five MRSA and thirty MSSA isolates were collected from the Second Affiliated Hospital of Zhejiang University School of Medicine as the experiment group .Twelve MRSA and twenty-two MSSA clinical strains were isola-ted as the control .All strains were identified by MALDI-TOF MS and the results were further analyzed by ClinProTools 3.0 software.Results Four algorithms including support vector machine (SVM), genetic al-gorithm ( GA) , supervised neural network ( SNN) and quick classifier ( QC) showed similar results in dis-tinguishing MRSA from MSSA isolates .The sensitivity of GA was 100 .0%and the sensitivities of other algo-rithms were all greater than 95.0%.The specificities of GA, SVM and QC were all greater than 90.0%. The areas under the receiver operating characteristic ( ROC) curves of four characteristic peaks at mass-to-charge ratios (m/z) of 3279, 6485, 6555 and 3299 m/z were all greater than 0.9.The virtual gel view showed that the bands generated by MSSA isolates at 3279 , 6485 and 6555 m/z were obviously deeper in color than those generated by MRSA isolates .However , the bands of MSSA isolates at 3299 m/z were appar-ently lighter in color than those of MRSA isolates .83.3%of MRSA and 90.0%of MSSA isolates from the control group were correctly identified by the GA model .Conclusion MALDI-TOF MS could rapidly and accurately identify MRSA from MSSA isolates under the strictly controlled experimental conditions with the advantages of less time-consuming, high sensitivity and high specificity .The accurate identification of MRSA from MSSA isolates could be applied for the prevention and treatment of MRSA infection .

BACKGROUND:It has been an urgent problem of how to promote cartilage repair of the knee and shorten the total course through a tissue engineering approach. Fortunately, microfracture plus stem cel transplantation may open up a new path for this issue. OBJECTIVE:To investigate the clinic feasibility of arthroscopic microfracture technique plus stem cel transplantation for repair of articular cartilage injury of the knee. METHODS:From October 2010 to March 2012, a total of 16 patients with articular cartilage injury of the knee were enrol ed, including 12 males and 4 females, with the average age of 38.6 years (16-52). Al cases of cartilage injury were confirmed by arthroscopy. Autologous bone marrow was extracted from patients at 2 weeks before treatment to isolate, culture and amplify bone marrow mesenchymal stem cel s in vitro. The cel culture solution of 3-5 mL (about 107 cel s) was harvested. The articular cavity was clean by arthroscopy and microfracture technique was performed at the area of cartilage injury that was then covered with hemostatic gauze through a minimal y invasive incision and the prepared bone marrow mesenchymal stem cel s were injected. The knee was bandaged with the elastic bandage after aspirating the joint cavity effusion by vacuum suction. Functional exercises were performed early by CPM.RESULTS AND CONCLUSION:After fol ow-up of 4-18 months, there were 13 cases of excel ent, 2 cases of valid and 1 case of ineffective. According to Lysholm knee scores, the average scores were improved from 42 points (33-67 points) to 89 points (75-99 points) at 4 weeks after treatment. The function was satisfied and al patients were fol owed up without recurrence or worse. Under the arthroscopy, the combination of microfracture technique and autologous bone marrow mesenchymal stem cel transplantation is proved to be effective for articular cartilage injury of the knee and it can notably improve the clinic symptoms and recover the function of the knee.

Objective:The study focuses on the distinct distributions of γδT cells in various tissues and the changes after Sal-monella typhimurium infection,and attempts to explore the physiological significance of γδT cell distribution and the role of γδT cells in infectious diseases .Methods:Flow cytometry and PCR technique were used to detect the proportion of different γδ T cell subsets among thymus,spleen,lymph nodes,liver,skin,and intestinal intraepithelial lymphocytes .Flow cytometry was applied to detect the secretion of IFN-γand IL-17a.The changes of various γδT cell subsets in liver and intestinal intraepithelial lymphocytes were analyze after Slamonella typhimurium infection.Res ults: γδ T cells were rich in the intestinal epithelium , skin and liver, but poor in the thymus,spleen and lymph nodes .The distribution of different subsets was quite dissimilar .Vγ5+γδT cells chiefly existed in skin ,and Vγ1+,Vγ4+,Vγ7+γδ T cells largely existed in small intestine.γδ T cells in liver mainly secreted IL-17a;however,γδ T cells in intestinal intraepithelial secreted IFN-γ.After infection by Salmonella typhimurium , the proportion of γδ T cells in intestinal intraepithelial increased significantly ,particularly Vγ1 +γδT cells.In Liver,there was no significant change of total γδT cell ratio,but the ratio of Vγ1 +γδT cells reduced ,Vγ4 +γδT cells raised.Conclusi on:γδT cells are rich in the intestinal epithelium ,skin and liv-er.The distribution of different subgroups has specificity .There are large differences in the ability of cytokine secretion among various subgroups of γδT cells.The distribution of γδT cell subgroups in small intestine and liver changes during Salmonella typhimurium in-fection.

Objective To introduce a new intelligent stomach syringe based on PC,which can deal with the stop of waste pipe automatically and feed back to the control of water speed.Methods On the foundation of analyzing the defects of traditional stomach syringe,the electric circuit of the design was improved.It realized the judgment,analysis and handling of acquired signals in the PC board.Results This machine can display the numerical value of pressure,PH,flux,and give the sound or light alarm.It can work automatically according to the demand or be operated artificially.Conclusion The instrument is automatic,secure and intelligent.Furthermore,it increases the successful rate for rescuing the patients.

Objective:To observe the protective effect of tremella polysaccharides (TP) on mouse aging model induced by D-galactose. Methods: Forty male ICR mice were divided into 5 groups (Group A: treated with 800 mg/kg TP; Group B: treated by 400 mg/kg TP; Group C: treated by 200 mg/kg TP; Group D: aging control; Group E: normal control). Except for Group E,other groups were injected with D-galactose [120 mg/(kg?d)] for 60 d to establish subacute aging model. Group A,B and C were treated with 800,400 and 200 mg/kg TP respectively via intragastric (i.g.) administration. Sixty days later,the spleen,heart, kidney and brain were collected to determine MDA, LP, SOD, GSH-Px and contents of hydroxyproline by colorimetry. Expression of P21 in the kidney were detected by immunohischemical technique. Proliferation and transformation of spleen lymphocytes induced by ConA were determined by MTT method. Results: Antioxidants SOD and GSH-Px activity of TP treated groups were higher than aging control group (P

0.05) between two groups.After EGDT,the level of cTnI in EGDT group was obviously dowered on the 7th day (0.16?0.22?g/L) and 14th day (0.09?0.13?g/L) compared with those in control group (0.39?0.43,0.29?0.29?g/L,P

Background Graft rejection is a primary cause of corneal transplantation failure,especially in high-risk keratoplasty.How to extend the survival time of graft is a problem to be solved.Objective This study was to investigate the influence of immune tolerance on high-risk rat keratoplasty induced by donor bone marrowderived immature dendritic cells (imDCs) transfected by chemokine receptor 7 (CCR7) recombinant adenovirus (Ad).Methods Bone marrow-derived imDCs were isolated and cultured from femur marrow of one male Wistar donor rat.The cells were transfected using recombinant Ad vector with rat CCR7 gene and resuspended in 500 μl PBS containing 1％ fetal bovine serum with the cells 1 × 107.High-risk corneal transplantaion models were established using monolateral corneal alkali-burn method in 60 SD rat recipients, and then allograft keratoplasty was performed with the 30 Wistar rats as donors.The models were randomized into the PBS group,imDCs group,imDCs with blank Ad vector group and imDCs with Ad-CCR7 group following the corresponding solution injection via caudal vein on preoperative day 7 and postoperative day 3 respectively.The survival time of graft was evaluated under the slit lamp microscope once per day.On the 14th day after operation, corneal sections were prepared from 6 eyes of each group for the pathological examination,and the relative expression levels of T helper cell 1 (Th1)-related factors,interleukin-2 (IL-2) mRNA and interferon-γ(IFN-γ) mRNA,as well as Th2-related factors, IL-4 mRNA and IL-10 mRNA, were detected by reverse transcription PCR (RT-PCR).The use and care of animals complied with the ARVO statement Results The mean survival time of grafts was (10.44±1.88) , (16.00±2.18) , (15.11±2.03) and(23.67±2.83) days in the PBS group,imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group, respectively, with a significant difference among the 4 groups (F =53.005, P =0.000).Compared with the PBS group, the mean survival time of grafts was considerably extended in the imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group (t=5.220,4.385,12.423 ,all at P=0.000) ,and a remarkble prolongation of graft survival duration was seen in the imDCs with Ad-CCR7 group in comparison with the imDCs group and the imDCs with blank Ad group (t =7.204,8.039,both at P=0.000).The relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were significantly lower,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were significantly higher in the imDCs group,imDCs with blank Ad group and the imDCs with Ad-CCR7 group than those in the PBS group (all at P＜0.05).Compared with the imDCs group and the imDCs with blank Ad group, the relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were remarkably delined,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were remarkably elevated in the imDCs with Ad-CCR7 group (all at P =0.000).Conclusions Application of imDCs transfected with CCR7 recombinant Ad via vena caudalis can prolong the survival time of grafts after keratoplasty of SD rats probably by affecting the balance of Th1/Th2 cytokines.

Objective To investigate the effects of medicine androgen deprivation therapy(ADT)on quality of life in patients with prostate cancer.Methods A total of 42 consecutive advanced prostate cancer patients without any other anti-androgen medications after ADT[A subcutaneous depot injection of LHRH-agonist(Zoladex) was instituted every 28 days]were enrolled.Levels of serum testosterone and prostatic specific antigen(PSA)were obtained just prior to ADT and after ADT.The general and disease-specific health-related quality of life were assessed.Results The average testosterone level was less than 50 mg/L after medical ADT in 3 weeks.And PSA level declined dramatically in one month.Although there were no significant differences on physical discomfort and limitations to daily activities.Urinary obstruction symptoms after ADT were disappeared and the size of prostate were reduced after one month.The appetite and vigor were worsened.Overall health status and sexual function were significantly reduced.Conclusion The ADT could make serum testosterone and PSA decline dramatically in short time,and worsen some general health-related quality of life.

Objective To investigate whether early goal-directed therapy ( EGDT ) could lower the mortality rate in patients with severe sepsis and septic shock. Methods Articles with items sepsis, severe sepsis, septic shock, EGDT were retrieved from MEDLINE, EMBASE, Cochrane, Wanfang Data and CNKI. Inclusion criteria included randomized controlled trial, subjects concerning patients with severe sepsis or septic shock, endpoints with short-term mortality [ in-hospital, intensive care unit ( ICU ) or 28-day ] and long-term mortality ( 60-day or 90-day ). Related risk ( RR ) and 95% confidence interval ( 95%CI ) were used as indices to judge the difference in mortality rate between EGDT group and standard treatment group. RevMan 5.2 software was used for Meta analysis. Results There were 8 studies meeting inclusive criteria with a total of 4 853 patients. For patients with severe sepsis and septic shock, compared with the group with routine treatment, EGDT showed a decrease in the short-term mortality ( RR = 0.74, 95%CI=0.66-0.82, P<0.000 01 ), but did not decrease the long-term mortality ( RR=0.99, 95%CI=0.92-1.06, P=0.81 ). Conclusion EGDT strategy may decrease the short-term mortality in patients with severe sepsis and septic shock, but it showed no influence on the long-term mortality.