Objective To observe the effects of exogenous basic fibroblast growth factor (bFGF) on radiation-induced apoptosis of C17.2 neural stem cells(NSCs) and explore the relationship between bFGF and Hes1,RBP-JK protein.Methods The cell viability was assessed using the MTT assay.After the cells attach to the flasks they were irradiated by the linear accelerator.And 5 min later,different concentrations of bFGF in accordance with the experimental design and cultured cells 48 h.Extracted total protein of each group and Western blot analysis showed the changes of the Hes1,RBP-JK protein.Results Compared with the control group,irradiation group cell growth was inhibited,the OD of 0 ng/ml bFGF group was 0.61±0.81,the OD of 80 ng/ml bFGF group was 1.21±1.01 and the control group was 1.51± 1.13.Compared with the control group,all groups showed statistically significant difference (P＜ 0.05).Western blot analysis showed that with the increasing concentrations of bFGF,RBP-JK expression gradually decreased,but Hes1 expression gradually increased.Conclusion Exogenous basic fibroblast growth factor (bFGF) can inhibit apoptosis of C17.2 NSCs.bFGF can regulate Notch signaling pathway downstream proteins Hes1,RBP-JK expression and inhibit neural stem cell apoptosis which were irradiated.

Objective To analyze the clinical character of uveitis in second hospital ot Jilin university.Methods Retrospectively analyze the clinical data of uveitis patients referred to from Second Hospital of Jilin University from September 2009 to September 2014.According to anatomical location,the manifestation of these uveitis patients were divided into anterior uveitis,panuveitis,intermediate uveitis and posterior uveitis.To discuss the possible causes of these patients according to the general information and relevant clinical laboratory examinations results.Results There were 1215 cases in this study,which included 587 male,accounting for 48.31％;and 628 female,accounting for 51.69％.The ratio of male-tofemale was 0.93∶1.The range of the age of these patients was from 4 to 91 years old.The mean age of these patients at the onset of these disease was (41.43±14.20) years.Of the 1215 cases,40 male and 43 female were younger than 20 years.The ratio of male-to-female was 0.93∶ 1;412 male and 396 female were between 21 and 50 years old.The ratio of male-to-female was 1.04∶ 1;135 male 189 female were older than 50 years.The ratio of male-to-female was 0.71∶1.There were 572 cases of anterior uveitis,accounting for 47.08％;527 cases of panuveitis,accounting for 43.37％;52 cases of intermediate uveitis,accounting for 4.28％;64 cases of posterior uveitis,accounting for 5.27％.703 cases had etiological diagnosis according to the clinical character and the auxiliary results,accounting for 57.68％.Vogt-koyanagi Haradal (VKH) syndrome,ankylosing spondylitis associated with uveitis and Behcet' s disease were the common entity,accounting for 30.44％,19.77％ and 14.22％ respectively.Conclusions The mean age of these patients in this study was older,compared to other reports.Female patients were more than male,especially in these patients older than 50 years.VKH syndrome,ankylosing spondylitis associated with uveitis and Behcet's disease were the common entities.

Objective To investigate the effect of Glutamine (Gln) on heat stress-induced dysfunction of intestinal epithelial barrier. Methods Human intestinal epithelial Caco-2 cells were pre-incubated with Gln for 24h and then exposed to heat 43℃ for 1h. Cell counting kit-8 (CCK-8) was used to detect the cellular proliferation with various concentrations of Gln and choose an optimum concentration for subsequent experiments. The barrier integrity was measured by transepithelial electrical resistance (TEER) and horseradish peroxidase (HRP) permeability. Levels of tight junction protein occludin and ZO-1 were analyzed by Western blotting. Cytoskeleton using Coomassie blue staining was observed by microscopy. Results At 0.7mmol/L concentration, Gln showed the most effective cell proliferation compared with other concentration groups (P<0.05). Therefore, 0.7mmol/L Gln was used as effective concentration in following experiments. Gln attenuated the TEER decrease and impairment of intestinal permeability induced by heat exposure compared with 43℃ group (P<0.01). The expressions of occludin and ZO-1 were significantly elevated by pretreatment with Gln. The distortion of cytoskeleton was also effectively prevented. Conclusion 0.7mmol/L Gln is potentially beneficial for protecting against heat stress-induced permeability dysfunction and epithelial barrier damage.

BACKGROUND: Gene therapy is a popular topic in domestic and overseas studies on biological therapy for brain tumor.OBJECTIVE: By using a newly constructed eukaryotic expression vector of pCR3-TK, the effect of the HSV-TK/ACV system on the proliferative activity of human glioma cells was investigated.DESIGN: Experimental study based on cells.SETTING: Department of neurosurgery and department of oncology in a university hospital.MATERIALS: The study was conducted at the National Key Laboratory of Veterinary Biotechnology of Harbin Veterinary Research Institute from January to April in 2004. The eukaryotic expression vector of pCR3-TK was constructed by the author. The TJ905 strain was a gift from professor Pu Pei-yu, who worked in the Neurology Institute of Tianjin city. The nontransfected cells and the cells transfected with pCR3-Uni vector were set as controls.METHODS: By using Lipofectamine(a cationic liposome), the pCR3-Uni vector and the recombinant pCR3-TK plasmid(inserted with HSV-TK gene)were transfected into the human glioma cell strain-TJ905. Then the positive clones were picked out and were given ACV(50 mg/L) . Totally 72 hours later, the cover slips were collected and silver staining for nucleolus organizer regions(AgNORs) was performed.MAIN OUTCOME MEASURES: After the ACV treatment and AgNORs staining, the numbers of silver-stained granules in TJ905 cells with or without transfections were counted respectively.RESULTS: In those cells transfected with HSV-TK gene, after ACV treatment, a significant decreasing in proliferative activity could be observed, and the average numbers of the silver-stained granules in cells transfected with pCR3-Uni or pCR3-TK were 14.33 and 6.67 respectively( P ＜ 0.01).CONCLUSION: As an easy-to-operate method, AgNOR counting is helpful for the studies on the proliferative activity of cells and the investigations into the potential anti-tumor mechanism of the HSV-TK/ACV system.

Objective To investigate prognostic factors for bloodstream infection in adult patients. Methods Clinical data of 131 adult patients with positive blood cultures during January 2002 to December 2003 in the Hospital were collected and 91 cases of them were retrospectively analyzed to understand their pathogen species and prognostic factors for it.Results Blood samples from 91 patients were cultured positive,53 cases(58.2%)with gram-negative bacteria mainly including Escherichia coli,Salmonella spp. and Klebsiella pneumoniae,28(30.8%)with gram-positive bacteria,mainly including Staphylococcus aureus and coagnlase-negative Staphylococci,eight(8.8%)with fungi and two(2.2%)with multiple infections.Case fatality ratio in this group of patients with septicemia was 30.8% during their hospitalization,and that in those with Pseudomonas aeruginosa,Staphylococcus aureus,Stenotrophomonas maltophilia and E.coli with extended-spectrum beta-lactamase was over 50%.Case fatality ratio was associated with severity of sepsis(OR=1.15)and inappropriately initial empirical treatment with antibiotics (OR=6.77).Conclusions Pathogen causing bloodstream infection in adults were mainly gram-negative bacteria and severity of infection and inappropriate initial antibiotics treatment could increase their fatality.

AIM:To research the effects of lithium chloride on transforming growth factor beta (TGF-β) and connective tissue growth factor (CTGF) in cultured human Tenon capsule fibroblasts (HTFs) and explore its mechanism.METHODS:HTFs were cultured and identified by vimentin staining with immunofluorescence and the morphological characteristics.The experimental group was processed 48h with LiCl in concentration of 80mmol/L, the control group without LiCl.The mRNA expression of TGF-β and CTGF in two groups were analyzed with real-time fluorescent quantitative polymerase chain reaction (real time-qPCR) and the protein expression was detected with Western blot.RESULTS:The cultured HTFs expressed TGF-β and CTGF.The mRNA expression of TGF-β and CTGF significantly decreased compared with the control group(t=20.042, 14.995, P<0.05).the protein expression of TGF-β and CTGF also decreased significantly compared with the control group(t=46.058、12.452, P<0.05)CONCLUSION:The cultured HTFs can express TGF-β and CTGF in mRNA and proteins' level.LiCl can reduce the expression of TGF-β and CTGF both in gene and proteins' level.LiCl has the potential to modulate wound healing for glaucoma filtration surgery.

OBJECTIVETo observe the relationship between variation of IL-2, IL-4, IL-18 and IP10 mRNA expressions in peripheral blood and the occurrence of acute graft-versus-host disease (aGVHD), and investigate whether some cytokines combined expression profiles could improve the diagnostic accuracy of aGVHD.

METHODSA total of 58 patients who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) were enrolled for the study. Peripheral blood samples were collected at different time points after transplantation. The mRNA expression levels of 4 kinds of cytokines (IL-2, IL-4, IL-18, IP10) were measured by real-time quantitative PCR (RQ-PCR). The relationship between mRNA expression level and the occurrence of aGVHD was analyzed with clinical features.

RESULTSThe expression levels of IL-2 and IL-18 at the onset of aGVHD were much higher than those after engraftment, being 2.69-fold and 3.12-fold increase, respectively (P = 0.000 & P = 0.000). The expressions of IL-2 and IL-18 mRNAs were slightly increased in patients with infection, but not statistically significant (P = 0.208 & P = 0.123). There was a slight but not statistically significant decrease of IL-4 and IP10 mRNA expressions at the onset of aGVHD (P = 0.230 & P = 0.325). Either IL-2 or IL-18 expression level could diagnose aGVHD as an independent factor (P = 0.000 & P = 0.000). The multivariate logistic regression analysis showed that the main factors related to aGVHD were IL-2, IL-18 and IL-4 (β = 1.13, P = 0.068 & β = 1.339, P = 0.047 & β = -0.600, P = 0.008 respectively). A composite panel of these three cytokines produced a better model for the diagnosis of aGVHD (AUC: 0.862, 95%CI: 0.768 - 0.957, P = 0.000), and the sensitivity and specificity were 75.0% & 83.3% respectively.

CONCLUSIONThe diagnosis of aGVHD can be optimized with a composite cytokines panel.

Objective To detect levels of IL - 4 and INF - ? in induced sputum dynamically in children with mycnplasma pneamo-niae pneumonia( MPP), and to analyze the function of Th1 /Th2 cell factor immune response in the genesis and development of MPP, so as to evaluate the clinical value of induced sputum method in MPP research. Methods There were 38 cases who were diagnoses as MPP using 3% high osmotic pressure of saline water to ultrasonic atomizing inhalation for inducing sputum. ELISA was used to detect IL-4 and INF-?. Results The content of IL-4 in acute stage was higher than that in convalescence stage in induced sputum of MPP children. Severe stage was higher than mild stage. However, the comparison between acute and convalescence stage didn't have statistics difference in the content of INF-?, neither did the comparison between severe and mild stage. IL- 4/INF- ? in acute stage was higher than that in convalescence stage. Severe stage was higher than mild stage. In convalescence stage, the comparison of INF - ?, IL - 4, IL - 4/INF - ? between the severe and the mild didn' t have statistic significance. Conclusions IL-4 and INF - ? have participated in the monogenesis of MPP. The disequilibria of Th1 /Fh2 is existed in MPP and Th2 reaction predominates. So induced sputum analysis can be a better way to judge the light or heavy press degree of MPP practically, conveniently and sensitively.

AlM: To investigate the clinical efficacy of treatment of dacryocystitis combined with canalicular obstruction by endonasal endoscopic dacryocystorhinostomy with canalicular intubation.
METHODS: A total of 27 cases of dacryocystitis combined with canalicular obstruction were treated by endonasal endoscopic dacryocystorhinostomy with canalicular intubation. Canalicular obstruction was treated by laser under dacryoendoscopy and antegrade intubation.
RESULTS: For 27 cases, operations were successful, with no complications. All patients were followed up for 6mo, 25 were cured, 2 were effective with no failed. The cure rate was 93%.
CONCLUSlON:Treatment of dacryocystitis combined with canalicular obstruction by endonasal endoscopic dacryocystorhinostomy with canalicular intubation has clear field, minimal invasion, quick recovery, exact effect and less recurrence, so it is worthy of promotion.

Objective To explore the effect of chitosan on vascular smooth muscle cells inhibited proliferation from rabbit arteriovenous fistula and its mechanisms.Methods Established rabbit fistula model on carotid arteryinternal jugular vein.After 1 month cultured VSMCs with primary culture by tissue-pieces inoculation.Cultured VSMCs were divided into three groups:①normal control group.②FBS-treated group:cell were treated with 5％,10％,20％ for 48 h,respectively; established the model of rabbit VSMCs proliferation.③chitosan-treated group:VSMCs cultured with 20％ FBS were exposed to different doses of chitosan(10,100,500,1000,2000μg/ml) for 48 h.And VSMCs were treated for different time (0,12,24,48 h) with Chitosan 1000 μg/ml.Expression levels of PCNA and TLR4/ NF-κB were detected by Western blotting.RT-PCR were applied to measure the mRNA expression of PCNA and TLR4.The protein levels of TLR4 and NF-κB were detected by immunofluorescence.Results Compared with low concentration serum group,FBS-treated VSMCs exhibited a increase in mRNA and protein expression of PCNA and TLR4.FBS-induced protein expression of PCNA and TLR4/NF-κB were reduced by chitosan.Also mRNA expression of PCNA and TLR4 were reduced.They were dependent on concentration and time.In rabbit VSMCs TLR4 was mainly expressed in the cytoplasm and NF-κB expressed mainly in the nucleus.Compared with normal control group,TLR4 and NF-κB protein expression were significantly decreased by chitosan.Conclusion High concentration serum induced VSMCs proliferation.Chitosan can inhibit the proliferation of rabbit VSMCs.It is speculated that the mechanism may be related to the expression of TLR4 receptor activation,reducing expression of downstream factor MyD88 and NF-κB.It is suggest that chitosan can become potential new drugs of arteriovenous fistula prevention of intimal hyperplasia.