Objective To observe the effect of RNA interference binding immunoglobulin protein (BiP) on expression of bone markers and keytranscription factors in osteoblast exposed to fluoride.Methods MC3T3-E1 cells were used as osteoblast model in vitro.The cell viability was test with cell counting Kit-8 after cells were administrated with varying concentrations of fluoride [0.0 (control),0.1,1.0,2.0,4.0,8.0,16.0,20.0,32.0 and 64.0 mg/L] for different duration.Cells transfected with small interfering RNA (siRNA) BiP were exposed to fluoride (2.0,8.0 and 20.0 mg/L) for 2 days.Real-time PCR and Western blotting technique were used to determine the gene and protein levels of BiP.Meantime,the mRNA expression of bone markers and key transcription factors was investigated by real-time PCR.Results The difference of all viability in fluoride-dose groups was statistically significant exposed for 1,3,7 and 14 days (F =46.7,118.6,214.6,325.6,all P ＜ 0.05).Expression of BiP significantly increased in cells exposed to 20.0 mg/L compared to that of control (11.22 ± 3.25 vs.7.94 ± 1.31,P ＜ 0.05).The expression of alkaline phosphatase (ALP) elevated in cells exposed to 2.0 mg/L of fluoride (12.81 ± 3.62 vs.6.86 ± 2.13,P ＜ 0.01);conversely,it significantly reduced in cells exposed to 20.0 mg/L of fluoride (0.89 ± 0.17 vs.6.87 ± 2.14,P ＜ 0.01).Cells transfected with siRNA BiP significantly decreased the ALP expression in cells exposed to fluoride compared to that of cells only exposed to the same concentration of fluorine (12.81 ± 3.62 vs.7.43 ± 2.06;5.92 ± 2.38 vs.3.96 ± 0.21,all P ＜ 0.05).Cells transfected with siRNA BiP and administrated with 2.0 mg/L significantly reduced the osteocalcin expression (4.29 ± 0.99 vs.1.29 ± 0.86,P ＜ 0.01).Similarly,expression of runt-related transcription factor 2 (Runx2) significantly increased in cells exposed to 2.0 mg/L.However,expression of Runx2 significantly decreased in cells transfected with siRNA BiP and administrated with fluoride.Cells transfected with siRNA BiP significantly decreased the Runx2 expression in cells exposed to 2.0 mg/L and 20.0 mg/L compared to that of cells only exposed to the same concentration of fluorine (1.13 ± 0.22 vs.6.61 ± 0.48;0.02 ± 0.02 vs.1.50 ± 0.38,all P ＜ 0.01).As the downstream of Runx2,the expression of osterix in cells treated with different concentrations of fluoride was similar to that of Runx2.Conclusion BiP is not directly involved in the process of osteoblast differentiation induced by fluoride;instead,it affects the expression of bone markers and key transcription factors in osteoblast exposed to fluoride.

Objective: To observe the preventive effect of alkaline drinking water on hyperuricemia in mice. Methods: Sixty male SPF Kunming mice were randomly divided into six groups: pH 7.3, pH 8.0, pH 9.3 intervention groups, in which the mice were given water with pH values of 7.3±0.5, 8.0±0.5 and 9.3±0.6, respectively; the control group, model group and positive drug group ( with 2 g/L allopurinol ) were given double distilled water. Except for the control group, the mice in each group were given yeast by gavage (1.5 g/mL) for 13 days. On the 14th day, the mice were injected with 300 mg/kg potassium oxyzinate by intraperitoneal injection, and then fasted for 1 day. On the 16th day, serum uric acid, creatinine and urea nitrogen were detected, and renal tissues were stained to observe the morphology.The expression levels of neutrophil gelatinase-associated lipocalin ( NGAL ), tissue inhibitor of metalloproteinase 1( TIMP1 ), organic anion transporter 1 ( OAT1 ) and urate transporter 1 ( URAL-1 ) in renal tissues were determined bywestern blotting. The mRNA expression levels of URAL-1 and OAT1 were detected by real-time fluorescent quantita⁃tive polymerase chain reaction. Results: The level of serum uric acid was higher in the model group than in the control group and in the pH 9.3 intervention group (both P<0.05). The number and area of renal tubular lesions were less in the pH 9.3 intervention group than in the model group (all P<0.05). The relative expression levels of NGAL and URAT-1 proteins were lower in the pH 9.3 intervention group than in the model group, and the relative expression level of OAT1 protein was higher in the pH 9.3 intervention group than in the model group ( all P<0.05). The relativeexpression level of URAT-1 mRNA was lower in the pH 9.3 intervention group than in the model group, and the rela⁃tive expression level of OAT1 mRNA was higher in the pH 9.3 intervention group than in the model group ( all P<0.05 ). Conclusion Alkaline drinking water with pH value of 9.3±0.6 can effectively prevent hyperuricemia and acute kidney injury in mice.

Objective To investigate the influence of Shenlongjiannaotang on the function of learning and memory of D-glalactose induced aging mimetic rats,and the concentrations of norepinephrine(NE),dopamine(DA) and 5-hydroxy tryptamine(5-HT) in cerebrum.Methods 50 rats were randomly divided into 5 groups: normal control group,model group,and high/middle/low dosage prescription group of Shenlongjiannaotang.The memory function was tested with Morris water maze.The concentrations of NE,DA and 5-HT were detected with HPLC with fluorescence spetrometer.Results In the model group,the latency of model mice increased,and the correct number decreased significantly compared with normal group(P<0.05).After 6 weeks administering Shenlongjiannaotang,the latency decreased and the correct number increased significantly in high dosage group,compared with those of the model group(P<0.05).The contents of NE and DA reduced in model group,and Shenlongjiannaotang could increase NE and DA contents of rats cerebrum(P<0.05).Conclusion Shenlongjiannaotang could improve the learning and memory function of D-galactose induced aging rats,which may related with the increase of NE and DA contents in cerebral tissue.The effects depend on the dosage of Shenlongjiannaotang.

Objective To investigate the effect of Shenlongtang on the function of learning and memory of aging rats induced with D-glalactose. Methods 40 Wistar rats were dividied into control group, model group, treating group 1 (large dosage), treating group 2 (medial dosage), treating group 3 (lower dosage). The aging models were induced with D-galactose. The memory function was tested with passageway water maze. The activity of superoxide dismutase (SOD) and content of malodialdehyde (MDA) was measured. Results In the D-galactose model rats, the swimming time was prolonged significantly, and the correct number was decreased significantly compared with normal group (P<0.05). After Six weeks treating with Shenlongtang, the swimming time significantly shortened and the correct number increased significantly in large dosage group compared with the model group (P<0.05). The activity of SOD increased and the content of MDA decreased in large dosage group compared with that in the model group (P<0.05).Conclusion Shenlongtang can improve the learning and memory function of aging rats induced with D-glalactose and resist lipid peroxide in brain tissue.

Objective To investigate the expression of serum soluble cytotoxic T lymphocyte associated antigen 4 (sCTLA4), the association of sCTLA4 level with erythrocyte sedimentation rate (ESR) and C reactive protein (CRP), as well as its role in patients with Crohn's disease (CD). The relationship-1661A/G and -1722T/C polymorphisms of CTLA4 gene and between disease susceptibility and phenotype of CD was analyzed. Methods A total of 126 CD patients and 300 healthy controls were enrolled in the study. Serum sCTLA4 level was determined by enzyme-linked immunosorbent assay. The concentrations of ESR and CRP were analyzed by automatic ESR Analyzer SRS 100/Ⅱ and rate nephelometry, respectively. The polymorphisms of CTLA4-1661A/G and -1722 T/C were genotyped by DNA sequencing. Results Serum sCTLA4 level was higher in CD patients than in healthy controls [(18. 70±3. 72) ng/ml vs (1.72±0. 32) ng/ml, P＜0. 01)]. Among CD patients, sCTLA4 level was higher in patients with active disease when compared to those with inactive disease [(19.83±4.35) ng/ml vs (18. 02±3.14) ng/ml, P=0. 015)]. sCTLA4 level was positively correlated with ESR and CRP levels (r=0. 267, P=0. 003; r=0. 524 P ＜0.01, respectively). In CD patients, serum sCTLA4 level was significantly higher in those with stricturing disease behavior than that in those without stricturing and penetrating or with penetrating disease behavior (P= 0.021; P=0. 015, respectively). Detection of CTLA4 -1661A/G and -1722T/C polymorphisms showed no significant difference between CD patients and healthy controls. Conclusions The high level of serum sCTLA4 in CD patients is correlated with disease activity, CRP levels and disease behavior. It suggests that sCTLA4 may play an important role in pathogenesis of CD.

Objective:To evaluate the clinical value of percutaneous microwave coagulation therapy for peripheral non-small-cell lung cancer. Methods:We evaluated 35 patients with non-small-cell lung cancer who received percutaneous microwave coagulation therapy and 35 patients who received radiotherapy from March 2004 to September 2006;the patients were sex-matched, age-matched, and had the same pathology and clinical staging. Clinical effects were observed and assessed. Survival rate were calculated using the Kaplan-Meier method. The difference in survival rate between the two treatment methods was analyzed using a log-rank test. Results:The 1-year, 3-year, and 5-year survival rates for the microwave coagulation therapy group (71.4%, 40.0%, and 20.0%, respectively) were significantly higher than those for the radiation therapy (51.4%, 22.9%, and 11.4%, respectively) (P<0.05). Conclusion:Percutaneous microwave coagulation therapy is a minimally invasive, safe, and effective alternative for patients with peripheral non-small-cell lung cancer who cannot undergo routine surgery because of poor heart and lung function or fear of surgical trauma.

Objective To investigate the function of major histocompatibility complex (MHC)class Ⅰ-related gene (MIC) in ulcerative colitis (UC) pathogenesis. Methods The difference of MICA, M ICB and their ligand NKG2D genes expression in colonic mucosa tissue of 34 UC patients and 12 healthy people were determined by fluorescent real-time reverse-transcriptase polymerase chain reaction (RT-PCR) and the expression location of M ICA in colonic mucosa tissue was obtained by laser scanning confocal microscope. Results The mRNA level of MICA, MICB and NKG2D expression in UC groups (3. 5408±2. 6658, 8. 9879±3. 2893 and 2. 4395±0. 8147 accordingly) was significantly higher than that in the healthy controls ( 1. 0477 ± 0. 7201, 4. 6293 ± 1. 2616 and 1. 1624±0. 3954 accordingly) (P = 0.0053, 0.0039 and 0. 0078 accordingly). It suggested that MICA was expressed in colonic epithelia cell membranes by laser scanning confocal microscopy.Conclusion The mRNA level of MICA, M ICB, and their ligand, NKG2D expression were all up regulated in the colonic mucosa of UC patients, which indicated MIC gene might perform important local function in UC.

Objective To develop an UPLC-MS/MS method for simultaneously determination of five components (adenosine,cytidine,guanosine,mannitol and adenine) in Qiangshenpaidu capsules. Methods The UPLC separation was performed on an Agilent ZOBAX SB-C18(2.1 mm×150 mm,5 μm) column.Isocratic elution was carried out with mobile phase consisting of methanol-0.1%formic acid (5∶95) at a flow rate of 0.2 mL.min-1.The mass spectrometer was operated in the positive ionization electrospray ( ESI) mode using multiple monitoring ( MRM) for analysis of five components. Results Adenosine,cytidine,guanosine,mannitol and adenine were all analyzed with good precision and accuracy. The linear ranges were 35-1 120 ng.mL-1( r=0.998 1) ,10-320 ng.mL-1( r=0.996 4) , 30-980 ng.mL-1( r=0.999 3) , 40-1 280 ng.mL-1( r=0.993 4), 25-800 ng.mL-1(r=0.996 5),respectively.The recoveries of six analytes ranged from 97.4% to 103.6% and the relative standard deviations were all below 4.7%. Conclusion A sensitive,accurate and suitable UPLC-MS/MS method has been developed,and the method could be applied for the determination of adenosine,cytidine,guanosine,mannitol,and adenine in Qiangshenpaidu Capsules.

Medical education should combine clinical professional skills with humanities skills,integrate humanities knowledge into vocational education.Taking 2008 grade seven-year program clinical medicine students in the 2nd affiliated hospital of Harbin Medical University as fostering object,we made researches into students' understanding of doctor-patient relationship before practice and their mastering of skills.Through conducting questionnaire,we got to know the effect of humanities practice skill training for seven-year program clinical medicine students.Meanwhile,we compared students' self evaluation results before and after training,discussed on how to improve medical students' communication skills,cultural skills and the reform direction in an aim to guide students to transit from students to clinical doctor.

Objective: To observe the expression of hMLH1 protein in esophagus squamous cell carcinoma, atypical hyperplasia tissue and normal esophagus tissue, so as to discuss the relationship between hMLH1 expression and esophagus carcinogenesis. Methods: The specimens of esophagus squamous cell carcinoma, atypical hyperplasia and normal esophagus tissue were obtained from 92 esophagus carcinoma patients. Immunohistochemistry (IHC) staining technique was used to detect expression of hMLH1 protein. The relationship between hMLH1 expression with clinical parameters, such as gender, age, cancer differentiation level, infiltration depth, tumor stage, lymphatic metastasis, was analyzed. Results: The positive rates of hMLH1 protein in esophagus squamous cell carcinoma, and atypical hyperplasia tissue and normal esophagus tissue were 36.96%,56.52%, and 84.78%,respectively,with the former 2 significantly lower than the latter (P