Objective: To evaluate the mechanism of T follicular helper cells ( Tfh ) in experimental autoimmune encephalomyelitis (EAE) via in vivo experiments. Methods:C57BL/6 mice were randomly divided into four groups,CFA group,EAE group,anti-ICOSL group and control group. Lymphocytes of different time points isolated from draining lymph nodes and spleen were stained for T follicular helper cells surface marker and T cells activation surface marker and analyzed by FACS. Observed parameters include inflammatory infiltration,demyelination in spinal cord and germinal center in spleen. ELISA was used to measure the level of antigen specific antibodies. Results: Mice in anti-ICOSL treated group developed mild disease was with lower clinical scores when compared with the EAE group. HE staining results turned out with alleviated inflammation and Luxol Fast Blue staining( LFB) showed no demyelization in anti-ICOSL treated mice compared with non-treated EAE models. Flow cytometry results revealed that percentages of T follicular helper cells decreased though the whole activated degree T cells was not influenced in anti-ICOSL treated group. Fewer ger minal center was found in both anti-ICOSL group and CFA group with reduced secretion of MOG-specific Ab. Conclusion:T follicular helper cells supported the development of cognate B cells,promoted the formation of germinal center,facilitate pathogenic MOG-specific Ab secretion,thus enhance EAE.

Objective To investigate the clinical value of serum procalcitonin (PCT) in diagnosis of urinary tract infection(UTI) in elderly patients.Methods 114 elderly patients with UTI in the department of infectious diseases of a hospital from January 2013 to December 2014 were analyzed retrospectively, clinical data of patients with abnormal and normal serum PCT were compared, PCT levels in patients with positive and negative blood cultures were compared, PCT receiver operating characteristic (ROC) curve for the diagnosis of bacteremia were drawn.Results Among 114 elderly patients with UTI, 46 were with abnormal PCT, 68 were with normal PCT.In abnormal PCT group, the proportions of patients with highest body temperature within 24 hours of admission, white blood cell count, neutrophil granulocyte percentage, C-reactive protein (CRP), blood urea nitrogen(BUN), creatinine(Cr), and urinary tract obstructive disease were all higher than those with normal PCT (all P<0.05).Among 42 patients with blood culture, PCT level in positive blood culture group(n=12) was higher than negative blood culture group(n=30)(1.93 [0.57-8.32] μg/L vs 0.36[0.15-1.01]μg/L, P=0.028).The area under the ROC curve (AUC) of the patients with bacteremia diagnosed by PCT was 0.72(95%CI:0.54-0.90),at the optimal value of 0.52 g/L, sensitivity, specificity, positive predictive value, and negative predictive value were 83.3%, 63.3%, 47.6%, and 90.5% respectively.Conclusion Serum PCT level can well reflect the severity of elderly patients with UTI, and is of great value in early diagnosis of bacteremia in elderly patients with UTI.

Objective To investigate the effect of apatinib on the proliferation,apoptosis and migration of pancreatic cancer cell line AsPC-1 in vitro.Methods Pancreatic cancer AsPC-1 cells were treated by apatinib in different concentrations.Cell proliferation and apoptosis were measured by CCK-8 and flow cytometry,and the effect of apatinib on cell migration ability was observed by wound healing assay.Results In control and 10,20,30,40 and 50umol/L apatinib treatment group,the inhibitory rates of AsPC-1 cells were 0,(1.45 ±0.68)％,(16.92±0.70)％,(23.84±0.84)％,(34.35±1.55)％ and (37.33± 0.81) ％,respectively.Cell proliferation was obviously inhibited by apatinib as the concentration increased,and the differences were statistically significant (P ＜ 0.05).In control and 20,40 umol/L apatinib treatment group,the apoptotic rates were (9.44 ± 0.18) ％,(16.62 ± 0.19) ％ and (25.42 ± 0.41) ％,respectively.Number of apoptotic cells was obviously increased by apatinib as the concentration increased,and the differences were statistically significant (P ＜ 0.05).In control and 20,40 umol/L apatinib treatment group,the migration ability was (29.5 ± 0.7) ％,(17.4 ± 0.9) ％ and (6.6 ± 0.5) ％,which was greatly decreased as the concentration increased,and the differences were statistically significant (P ＜ 0.05).Conclusions Apatinib can effectively inhibit the proliferation and migration of pancreatic cancer AsPC-1 cells and induce apoptosis.

Objective To observe the clinical effect of xerophthalmia of meibomian glands dysfunction treated with meibomian glands massage and tobramycin dexamethasone in the northeast area.Methods The clinical data of 403 patients (767 eyes) with xerophthalmia of meibomian glands dysfunction in the northeast area in January to November 2013 were analyzed retrospectively.The patients were divided into 3 groups.Patients of treatment goup 1 were combined modality therapy by meibomian glands massage and tobramycin dexamethasone and artificial tears (carbomer eye ophthalmic gel).Patients of treatment goup 2 were treated with tobramycin dexamethasone and artificial tears.Patients of control group were only used by artificial tears.Tear film break-uptime (BUT),tear secretion test (Schirmer Ⅱ),corneal staining score and symptom score were measured before treatment and after treatment for 1,3 months.Results There was no significant difference in the subjective symptoms,BUT,Schirmer Ⅱ,corneal staining score before treatment among three groups (P ＞ 0.05).Compared with that before treatment,the subjective symptoms,BUT,Schirmer Ⅱ,corneal staining score after treatment for 1 month was improved in treatment group 1 and treatment group 2 (P ＜ 0.01),but there was no significant difference in control group (P ＞ 0.05).After treatment for 3 months,the effective power in treatment group 1 [87.1％(122/140)] and treatment group 2 [60.8％(79/130)] was higher than that in control group [48.9％(65/133)],and there was significant difference (P ＜ 0.05).And there was significant difference between treatment group 1 and treatment group 2 (P ＜ 0.05).Conclusions Xerophthalmia of meibomian glands dysfunction in the northeast area due to speciality in the geographical environment and food habits.Combined modality therapy (applied heating,meibomian glands massage and tobradex) can provide a new direction of xerophthalmia of meibomian glands dysfunction,retrieve in the lipids component of the tear film and eliminate the inflammation.But,dependence of the out-patients are very important in the therapeutic process.

Objective To establish standardized traceable management procedure for implanted high-value consumables in operating room.Methods The management model combining information-based system operation process and quality control process was designed,and management results before and after implementation were compared.Results There were statistically significant differences in error rates of information recording,bar code sticking and charging of implantable high-value consumables after the implementation of the process management mode (P＜0.05).At the same time,there were statistically significant differences in improvement of traceability of high-value consumables,adverse event reporting and patient satisfaction(P＜0.05).Conclusion Establishment of management model in operating room for implanted high-value consumables can ensure medical safety and increase medical quality.It was proved to improve the level of hospital management.

Objective To investigate the effect of the expression of Notch1 protein and the mutation of Notch1 gene in.T-cell lymphoma (TCL).Methods Immunohistochemistry was used to detect the expression of Notch1 protein,and PCR amplification and DNA sequencing were used to detect the mutation of Notch1 gene in the 26th and 27th HD domain and the 34th PEST domain in 30 cases.10 cases of reactive hyperplasia tissues of lymph node were as the control.Results The positive rates of Notch1 protein expression and Notch1 gene mutation were 70.0 ％ (21/30) and 56.7 ％ (17/30).8 cases of Notch1 mutations were detected in the HD domain,6 cases in the PEST domain,and 3 case in both HD and PEST domains.Inscrtion,deletion,nonsense mutation and missense mutation were included in Notch 1 mutations.Conclusion Notch1 gene mutation may play an important role in the expression of Notch1 protein.The occur of TCL is related to the expression of Notch1 protein and the mutation of Notch1 gene.

Objective To investigate the effect and mechanism of chitosan on vascular smooth muscle cell proliferation of uremia patients with arteriovenous fistula.Methods Primarily culturing the VSMCs of uremia patients with arteriovenous fistula and patients without uremia by explants adherent method,and taking the second generation.VSMCs from patients without uremia cultured with 20％ FBS medium were non-uremia group,VSMCs of uremia patients cultured with 20％ FBS medium were uremia group,VSMCs of uremia patients with 100 pg/ml chitosan were uremia+ chitosan group.The expression of α-SMA was detected by immunohistochemistry.The changes of migration and invasion of VSMCs were detected by scratches and transwell migration assays.The mRNA expressions of TLR4 and PCNA were measured by real-time PCR.VSMCs of uremia patients with arteriovenous fistula were intervened with different doses of chitosan (0,100 and 500 μg/ml),and the protein expressions of TLR4,MyD88 and NF-κB were detected by Western blotting.Results Compared with those in non-uremia group,in uremia group and uremia+chitosan group α-SMA was upregulated,migration and invasion of VSMCs were enhanced,and mRNA expressions of TLR4 and PCNA were increased (all P ＜ 0.05).Compared with those in uremia group,the level of α-SMA was significantly decreased,the ability of migration and invasion of VSMCs were decreased,and the mRNA expressions of TLR4 and PCNA were decreased (all P ＜ 0.05).TLR4,MyD88 and NF-κB protein expressions were reduced in concentration-dependent manner by 100 and 500 μg/ml chitosan.Conclusions (1) In vitro,chitosan decreases the ability of migration and invasion of VSMCs of uremia patients with arteriovenous fistula.(2) Chitosan inhibits the proliferation of VSMCs,which may be relevant in the decreased expressions of TLR4,MyD88 and NF-κB.

Objective To study the N terminal pro brain natriuretic peptide (NT-proBNP)in serum of hypertension patients. Methods 152 patients with hypertension were included in this study.According to the different degree and development of hypertension,152 patients were divided into three different grade group of hypertension:first grade of hypertension group (30 patients),second grade of hypertension group (36 patients)and third grade of hypertension group (86 patients).And the 86 patients were divided into hypertension group (40 patients)and hypertension with diabetes group (46 patients)re-spectively,comparaed with the level of NT-proBNP in serum of different hypertension grade groups.Results The levels of the NT-proBNP in serum were 68±44,122±31 and 834±309 pg/ml of first grade of hypertension group,second grade of hypertension group and third grade of hypertension group,respectively.The level of the NT-proBNP was gradually increased with grade of hypertension (t=2.455,3.561,P<0.01).The level of the NT-proBNP (1 178±864 pg/ml)of hypertension with diabetes group was significantly higher than hypertension group (599±411 pg/ml)(t=3.785,P<0.01).Conclusion The level of NT-proBNP can obj ectively reflect the grade of the disease in patients with hypertension.it is a certain signifi-cance guiding for monitoring the clinical treatment of the disease.

Objective To investigate antimicrobial resistance and ceftriaxone resistance mechanisms in clinically isolated nontyphoidal Salmonella(NTS),and provide evidence for the prevention and control of NTS infection and rational use of antimicrobial agents.Methods 108 NTS isolates were isolated from stool specimens of outpatients with acute diarrhea in the Second Hospital of Tianjin Medical University and Tianjin Medical University General Hospital from May to October of 2014,NTS were performed antimicrobial susceptibility testing;non-ceftriaxone-susceptible isolates were typed by serological,multilocus sequence (MLST),and pulsed-field gel electrophoresis (PFGE)methods,extended-spectrumβ-lactamase (ESBL)detection and AmpC genes were detection.Results Among 108 NTS isolates,mono-drug resistance rate to 11 antimicrobial agents was 49.07% (n= 53),multidrug resistance rate was17.59% . Susceptibility rates to nalidixic acid,levofloxacin,ciprofloxacin,ceftriaxone,and ertapenem were 61.11% ,66.67% ,68.52% ,97.22% ,and 100.00% respectively. Three non-ceftriaxone-susceptible NTS isolates were detected,2 were ST11 Salmonellaenterica serotype (Sa8709,Sa8771),1 was ST34 Salmonellatyphimurium serotype(Sa8763). Cluster analysis of PFGE revealed that Sa8709 was highly similar to Sa8771 strains(91 .70% ), but the similarity to Sa8763 was low(55.80% );Sa8709 strain carried CTX-M gene,Sa8771 strain carried CTX-M and TEM genes,Sa8763 strain carried OXA gene. Conclusion Clinically isolated NTS in this area are low resistant to fluoroquinolones,multidrug resistant strains carrying ESBLs have emerged.

Objective To silence the beclin1 gene expression by using RNA interference technology in AR42J rat pancreatic acinar cells,and build a stable AR42J line silencing beclin1.Methods Three kinds of shRNA targeting rat beclin1 mRNA and negative control shRNA were designed and synthesized,and were inserted into the plasmids GV112,respectively.The recombinant plasmids were named as p-sh-Beclin1-1,psh-Beclin1-2,p-sh-Beclin1-3 and p-shRNA-NC.Lipo3000 was used to transfect the recombinant plasmid into AR42J cells,the expression of beclin1 mRNA were detected by RT-PCR to screen for the most efficient silencing plasmid,and then it was packaged into lentiviral (LV).AR42J cells were infected with LV and screened by puromycin.Beclin1 mRNA and protein expression was determined by RT-PCR and Western blot.Results The recombinant plasmid was confirmed by agarose gel electrophoresis and sequencing showed that shRNA sequences were in line with expectations.The beclin1 mRNA inhibition rates of AR42J cells after p-sh-Beclin1-1,p-sh-Beclin1-2,p-sh-Beclin1-3 and p-shRNA-NC transfection were (17.8 ± 4.0) ％,(30.6 ± 2.8) ％,(45.8 ± 7.7) ％,(7.0 ± 11.8) ％,respectively.The inhibition rates of three p-sh-Beclin1 transfection cells were significantly higher than that in non-transfection cells,and the difference was statistically significant (P ＜ 0.05).While the inhibition rate of p-shRNA-NC transfection cells was not significantly different from that of non-transfection cells,p-sh-Beclin 1-3 with highest rate of inhibition was packaged by LV,and infected AR42J cells,then puromycin was applied to screen,inhibition rate of beclin mRNA expression in LV infection cells was (86.1 ± 1.2) ％,and the protein expression inhibition rate was (87.9 ± 2.8) ％,and the difference between infection and non-infection groups was statistically significant (P ＜ 0.05).Conclusions The stable AR42J line silencing beclin1 is successfully established,which can provide a new cell model for future research of the role of beclin1 in the pathogenesis of acute pancreatitis.