To isolate the specific genes in protoscoleses (PSC) of Echinococcus granulosus under oxidative stresses from the SSH library constructed in the previous study, the gene expression in PSC under oxidative stresses was studied by using real-time PCR. The previously amplified library was sequenced and analyzed in GenBank with Blast research. Sequence analysis indicated that all clones in the SSH library contained the coding sequences, of which some clones showed homology in the GenBank and others were unknown. Differential expression of 4 genes randomly selected and the TPx gene in this library were studied with real-time PCR. It was demonstrated that the gene expression of S88 and H32-1 in oxidative tissues was 2.0 and 2.3 times higher than the un-oxidative stresses respectively. The TPx gene was up-regulated when PSC was induced with H_2H_2 of more than 0.8 mmol/L. These results implies that the up-regulated expression of the above-mentioned genes may be related with the related functions of anti-oxidative process in PSC and they may be used as the candidate genes for the study of anti-oxidation of E.granulosus.

Objective To screen the potent permeation enhancers used in transcutaneous immunization with inactivated highly pathogenic avian influenza vaccine. Methods Five different permeation enhancers, ethanol, propylene glycol, dimethyl sulfoxide, ratinoic acid, oleic acid, were used to treat the skin of BALB/c mice before transcutaneous immunization. Sera were collected before the flist transcutaneous immunization and every two weeks post immunization. The titers of influenza virus-specific humoral IgG and IgA were assayed in serum, lung and nasal lavages by ELISA. The titers of hemagglutination inhibition ( HAI), IFN-γand IL-4 produced by splenic lymphocytes were also detected. Except that, clinical symptom of the skin in different time points and skin pathological changes were observed. Results The serum IgG titers, HAI titers and the influenza virus-specific lgA and IgG in lung and nasal lavages in the groups of HA +CT + DMSO, HA + CT + RA and HA + CT + OA were significantly higher than those of HA and HA + CT groups( P ＜0.05). Moreover, the numbers of splenic lymphocytes producing IFN-γ and IL-4 were increased in the above three groups than those in control groups. In addition, no evident clinical symptoms were observed, but stratum corneum of the skin in different groups showed different changes. Conclusion DMSO,RA and OA are potent permeation enhancers in mouse model inoculated with inactivated high pathogenic avian influenza vaccine transcutaneously.

The main impacting factors on implementing clinical pathway were analyzed by applying fishbone diagram.The factors include policy factors,organizational factors and personal factors.It was suggested to improve the trial work of implementing clinical pathway management by reforming the medical care payment system,putting the clinical pathway management into the evaluation system,building up medical information system,putting more efforts on promotion and enhancing cooperation among related departments.

Objective To explore the association between X-ray repair cross complementing group 1 (XRCC1)gene rs25487 locus polymorphisms and nonobstructive azoospermia in Hui minority ethic population of Shaanxi Province.Methods We used polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)method for genotyping at XRCC1 gene rs25487 locus in 79 patients with nonobstructive azoospermia and 82 healthy male controls in Hui minority ethic population of Shaanxi Province.Then we analyzed the association be-tween XRCC1 gene rs25487 locus and nonobstructive azoospermia.Results Compared with GG genotype,GA, AA and GA + AA genotypes demonstrated a significantly increased risk for nonobstructive azoospermia (OR =2.286,95% CI 1.1 5 1-4.539;OR =2.202,95% CI 0.753-6.439;OR =2.271,95% CI 1.1 71-4.403),respec-tively.Meanwhile,the A allele frequency was significantly higher in azoospermic patients than in controls (OR =1.582,95% CI 1.005-2.492,P =0.047).Conclusion G→A in XRCC1 gene rs25487 locus is correlated with nonobstructive azoospermia in Hui minority ethic population of Shaanxi province.

Objective To establish a stable in vitro model of blood-brain barrier (BBB) simulating in vivo state using the primary-cultured rat brain microvascular endothelial cells (BMVECs) and pericytes.Methods The primary rat BMVECs and pericytes were isolated,purified and cultured.The isolated cells were identified by immunocytochemical staining method.An in vitro model of BBB was constructed using Transwell inserts (pore size 0.4 μm) coculture.Its barrier function was evaluated by the 4-hour leakage test,tight junction protein identification,transendothelial resistance detection,and permeability test.The difference between the cocultured model and simple BMVEC model across the membrane resistance values,and the permeability difference of the small molecule sodium fluorescein (Na-F) were compared.Results Confluent BMVEC monolayers demonstrated a typical cobblestone appearance and the pericytes displayed irregular shape and overlapping growth.Immunodouble labeling technique identification showed that the pericytes positively expressed α-srmooth muscle actin (α-SMA) and neuron-glial antigen 2 (NG2); after the fusion of cocultured model endothelial cells,the surface leakage test became positive; immnocytochemical staining shows that a continuous and dense tight junction formed between the endothelial cells; compared to the BMVEC model,the transendothelial electrical resistance of the cocultured model increased significantly (190.762 ± 10.326 Ω/cm2 vs.96.503 ± 8.012 Ω/cm2; t=- 24.489,P ＜0.01),and the permeability decreased significantly (56.149％ ± 3.572％ of the single endothelial model; t =19.330,P ＜ 0.01 ).Conclusions The primary isolated rat BMVECs and pericytes cocultured the morphology,structure and barrier function of in vitro model have the basic characteristics of BBB,and they have provided a useful tool for the research of BBB.

Severe acute respiratory syndrome is an infectious disease caused by a new type of coronavirus. It belongs to the seasonal febrile diseases in traditional Chinese medicine. The prevention and treatment of severe acute respiratory syndrome (SARS) can be under the guidance of the doctrines for treating febrile diseases of traditional Chinese medicine, treatment based on syndrome differentiation, such as syndrome differentiation of triple energizer, syndrome differentiation according to defensive phase, qi phase, nutrient phase and blood phase. During April and May of 2003, 8 cases of SARS were diagnosed in Shanghai, and 6 patients accepted complementary therapy of traditional Chinese medicine, without death case. The only one patient who didn't take glucocorticoid therapy was complementarily treated with traditional Chinese herbs through the whole treating procedure. Upon the successful treatment of the eight cases of SARS in Shanghai, it is demonstrated that the triple-energizer syndrome differentiation and defensive-qi-nutrient-blood syndrome differentiation in traditional Chinese medicine are of high value in treating SARS patients.

Objective To investigate the effects of β-blockers on cardiac protection and hemodynamic in patients with septic shock. Methods A prospective randomized controlled trial was conducted. Forty-one patients with septic shock in accordance with early goal directed treatment and met the target within 6 hours,and admitted to intensive care unit (ICU)of Affiliated Huxi Hospital of Jining Medical College from January 2012 to January 2014 were enrolled. The patients were divided into treatment group (n=21)and control group (n=20)by random number table. The patients in both groups were given the standard treatment,esmolol was giving to patients in treatment group in order to control the heart rate (HR)below 100 bpm within 2 hours,and the patients in control group only received standard treatment. The changes in hemodynamic parameters〔mean arterial pressure(MAP),central venous pressure(CVP), HR,cardiac index(CI),stroke volume index(SVI),systemic vascular resistance(SVRI),global end diastolic volume index(GEDVI)〕,biochemistry metabolic of tissue〔central venous oxygen saturation(ScvO2),lactic acid(Lac)〕,and cardiac markers 〔troponin I (cTnI)〕before and 12,24,48,72 hours after the treatment were recorded. Results①Before treatment,the hemodynamic parameters,tissue metabolism index and cTnI had no significant differences in both groups (all P>0.05).②The hemodynamic parameters after treatment in the control group showed no significant difference compared with that before treatment. HR and CI in the treatment group were gradually declined after treatment,SVRI and GEDVI were gradually increased. There were significant differences in HR,CI,SVRI,and GEDVI between treatment group and control group from 12 hours on〔HR(bpm):93±4 vs. 118±13,CI (L·min-1·m-2):3.3 ±0.8 vs. 4.5 ±0.6,SVRI (kPa·s·L-1·m-2):159.2 ±27.4 vs. 130.5 ±24.2,GEDVI(mL/m2):668 ±148 vs. 588 ±103,P<0.05 or P<0.01〕. MAP,CVP and SVI in the treatment group showed no significant changes. ③Lac after treatment in both groups was decreased slowly,Lac (mmol/L)at 12 hours after treatment was significantly decreased compared with that before treatment (control group:8.8 ±3.2 vs. 9.8 ±3.4,treatment group:9.5±3.1 vs. 10.5±4.1,both P<0.05). The Lac of control group and treatment group were 2.5±1.2 and 2.7±1.1 at 72 hours after treatment,and there was no significant difference between two groups (all P>0.05). The ScvO2 was not decreased in both groups.④Compared with before treatment,cTnI in the control group was gradually increased,peaked at 72 hours,and that in the treatment group was gradually increased,peaked at 24 hours and then gradually declined. Compared with control group,the cTnI (μg/L)in the treatment group was decreased significantly at 24,48,72 hours (1.15 ±0.57 vs. 1.74 ±0.77,0.93 ±0.52 vs. 2.15 ±1.23,0.52 ±0.36 vs. 2.39 ±1.17,all P<0.01). Conclusionsβ-blockers (esmolol) can improve cardiac function and myocardial compliance,reduce the myocardial injury in patients with sepsis shock. Although β-blockers can decrease cardiac output,it has no influence on the circulation function and tissue perfusion.

Objective To explore the clinical value of transnasal high volume oxygen therapy in the treatment of patients with acute heart failure.Methods From January 2016 to January 2018,61 cases with acute heart failure in Huxi Hospital Affiliated to Jining Medical College were selected.The patients were randomly divided into control group and treatment group according to the digital table,31 cases in the control group and 30 cases in the treatment group.The two groups were routinely given control of fluid volume,analgesia,strong heart,diuresis,vasodilator,anti-platelet aggregation,camp support and so on.The control group was given conventional oxygen therapy,and the treatment group was treated with high flow oxygen through nose.Before treatment and 12h,24h,48h,72h after treatment,the left ventricular ejection fraction (LVEF),oxygen index (PaO2/FiO2),serum lactic acid (Lac),B type sodium and titanium (BNP) in serum,and the application rate of non-invasive mechanical ventilation and invasive mechanical ventilation in 7d were observed in the two groups.Results Compared with the control group,the LVEF of the treatment group in each time point increased [(35.58 ± 3.64) ％ vs.(37.77 ± 3.76) ％,(37.87 ± 3.58) ％ vs.(40.07 ±3.36)％,(44.94 ±3.19)％ vs.(46.83 ±3.21)％,(47.55 ±3.45％)％ vs.(40.07 ±3.36％)％,t =-2.308,-2.466,-2.316,-2.487,all P ＜ 0.05].The PaO2/FiO2 of the treatment group increased significantly at each time point after treatment [(177.39 ± 10.62) mmHg vs.(184.17 ± 10.49) mmHg,(188.00 ± 11.72) mmHg vs.(198.57 ± 18.47) mmHg,(204.06 ± 17.69) mmHg vs.(221.40 ± 23.80) mmHg,(265.23 ± 34.51) mmHg vs.(290.37 ± 26.72) mmHg,t =-2.507,-2.678,-3.236,-3.174,all P ＜ 0.05].The BNP level of the treatment group decreased significantly at each time point after treatment [(2 462.90 ± 288.00) ng/mL vs.(2 264.53 ± 366.44) ng/mL,(1 646.61 ± 377.19) ng/mL vs.(1 474.07 ± 214.03) ng/mL,(991.94 ± 242.95) ng/mL vs.(811.90 ±258.67) ng/mL,(653.77 ± 147.671) ng/mL vs.(526.47 ± 127.87) ng/mL,t =2.355,2.187,2.803,3.594,all P ＜ 0.05].The Lac level of the treatment group decreased significantly at 12h and 24h after treatment [(5.05 ± 0.69) mmol/L vs.(4.55 ± 0.80) mmol/L,(3.68 ± 0.89) mmol/L vs.(3.13 ± 0.77) mmol/L,t =2.610,2.601,all P ＜ 0.05],but there were no statistically significant differences between the two groups at 48h and 72h after treatment [(1.62 ± 0.65) mmol/L vs.(1.53 ± 0.65) mmol/L,(1.36 ± 0.64) mmol/L vs.(1.26 ± 0.46) mmol/L,all P ＞ 0.05].In the control group and the treatment group,the incidence rates of non-invasive mechanical ventilation in 7d were 35.48％ (11/31),13.33％ (4/30),respetively,the difference was statistically significant (x2 =4.034,P ＜ 0.05).In the control group and the treatment group,the incidence rates of invasive mechanical ventilation in 7d were 12.90％ (4/31),3.33％ (1/30),respetively,the difference was statistically significant (x2 =4.957,P ＜ 0.05).Conclusion Nasal high flow oxygen therapy has better clinical effect on patients with acute heart failure.It is a more active treatment measure,and is worthy of clinical application.

The main work and achievements of clinical pathway work in China since 2009 were systematically reviewed in the paper. It analyzed the problems existing in the implementation of clinical pathway management in China, and suggested on such management in the future. The suggestions include:deeper understanding, convergence with the payment system reform, strengthened quality control, further informatization,and better performance appraisal system.

OBJECTIVETo explore the effect of recombinant human soluble CD(40) ligand (rhsCD(40)L) and CD(40)L cDNA transfected cell (CD(40)L-TC) on the behavior of malignant B lymphocytes, and investigate the possibility of using rhsCD(40)L as a new bio-factor in tumor immunotherapy.

METHODrhsCD(40)L and CD(40)L-TC were obtained by gene recombinant techniques. Multiple myeloma cell lines, XG2, XG7, U266 and 8226, B-lymphoma cell lines, Raji and Daudi were selected to detect responses to rhsCD(40)L and CD(40)L-TC stimulation. Cell growth curve, cell cycle, early apoptosis as well as membrane surface molecules on these cell lines were analyzed.

RESULTS(1) The expression levels of CD(40) molecule on malignant B lymphocytes showed heterogeneity. High level of CD(40) on XG2, moderate on 8266, Raji, and Daudi, and no expression on U266 and XG7 were detected. The rhsCD(40)L stimulation gave rise to a typical homo-type cell aggregation of XG2 and Daudi. Meanwhile, at least 10 to 20 of CD(40)(+) XG2 or CD(40)(+) Daudi cells were found adherent to one pre-treat ed CD(40)L-TC. (2) Co-incubation with rhsCD(40)L (5 micro g/ml), or CD(40)L-TC (tumor cell: CD(40) = 5:1) resulted in a significant inhibition of in vitro cell growth of XG2, Raji and Daudi, with G(1)-phase arrest for XG2 and G(2)-phase for Raji and Daudi. These two kinds of CD(40) stimulators induced XG2, Raji and Daudi cells to apoptosis in vitro. The apoptotic rate for XG2 was 23.3% (rhsCD(40)L) and 18.8% (CD(40)L-TC), for Daudi 14.2% and 15.9%, and for Raji 11.6% and 8.9% respectively. (3) Phenotype analysis showed that CD(95) expression levels were significantly up-regulated on XG2, Raji and Daudi after stimulation with rhsCD(40)L or CD(40)L-TC, and CD(80) and CD(18) expression levels on Raji were respectively enhanced and decreased.

CONCLUSIONThe abilities to directly inhibit XG2, Daudi and Raji cell proliferation, to induce themapoptosis, as well as to up-regulate immune co-stimulator molecule CD(80) expression on Raji cells would make rhsCD(40)L a potential bio-factor for tumor immuno-therapy.

B-Lymphocytes ; drug effects ; metabolism ; pathology ; CD40 Antigens ; metabolism ; CD40 Ligand ; genetics ; metabolism ; pharmacology ; Cell Division ; drug effects ; Coculture Techniques ; DNA, Complementary ; genetics ; Humans ; Lymphoma, B-Cell ; metabolism ; pathology ; Recombinant Proteins ; pharmacology ; Time Factors ; Transfection ; Tumor Cells, Cultured ; drug effects