Objective To evaluate the biological effect of tumor necrosis factor-α(TNF-α) on the proliferation and multi-directional differentiation of stem cells from rat apical papilla(SCAP).Methods SCAP was extracted by combining enzyme digestion method with tissue block method.The cells were divided into control group(TNF-α 0 ng/mL) and experimental group(TNF-α 5,10,20,50 ng/mL).The ability of proliferation of SCAP was measured by MTT method.The ability of osteogenic/dentinogenic differentiation of SCAP was measured by alizarin red staining and quantitative real-time PCR.The ability of adipogenic of SCAP was measured by oil red O staining.The expression of vascular related genes of SCAP was measured by quantitative real-time PCR.Results SCAP was consistent with the characteristics of mesenchymal stem cells and possessed the ability of multi-directional differentiation.The MTT results showed that experimental group promoted the proliferation of SCAP in comparison with the control group.The difference was statistically significant(P<0.05),and 10 ng/mL was the optimum concentration.The results of alizarin red staining showed that with the increase of the concentration of TNF-α,the mineralized nodules in the experimental group gradually became smaller,and the number of the formation decreased gradually.The results of quantitative real-time PCR showed that the expression of OC,DMP-1 and DSPP in the experimental group was significantly lower than that of the control group at 3 and 7 days,in which the expression of OC was statistically significant different(P<0.05);at 14 days,the expression of OC,DMP-1 in the experimental group was significantly lower than that of the control group(P<0.05).The result of Oil red O staining showed that with the increase of the concentration of TNF-α,the lipid droplets formation in the experimental group gradually decreased.The result of quantitative real-time PCR showed that the expression of ANGPT1,VEGFA,PECAM-1 in the experimental group was significantly lower than that of the control group(P<0.05).Conclusion TNF-α might promote the proliferation and inhibit the multi-directional differentiation of SCAP.

AIM To study the pharmacokinetics of arbidol capsule in Chinese healthy volunteers.METHODS A single oral dose of arbidol capsule 200 mg was given to 20 healthy volunteers respectively.Plasma samples were prepared based on a simple liquid-liquid extraction.The extracted samples were analyzed by HPLC equipped with UV detection.Pharmacokinetic parameters were calculated by 3P87 software. RESULTS The main pharmacokinetic parameters of arbidol were as follows:c(max)(418±s 241)μg·L-1,t(max)(1.3±1.2)h,t(1/2α)(1.9±2.3)h,t1/2β(14±5),hAU0-t(2 633±1 071)μg·L-1,Vc/F(0.7±0.6)L,CL(0.08±0.03)L·h-1,CONLUSION The pharmacokinetics of arbidol capsule in human body accord with two-compartmetn open model.The study will offer the pharmacokinetic parameters for the clinical application of arbidol.

OBJECTIVE:To study the stability of Urapidil injection separately mixing with KCl injection,potassium mag?nesium-L-aspartate injection,NaHCO 3 injection,vitamine C injection and lidocaine injection in10%glucose injec?tion.METHODS:Ultraviolet spectrophotometry was used for detecting the changes of absorbance,absorption curve,and pH at15℃～25℃within0～8hours,and the external appearance and changes under light microscope were observed.RESULTS:There were no evident changes in absorbance,absorption curve,external appearance,microscopic findings and pH.CONCLUSION:Urapidil injection is stable in mixing with above-mentioned5drugs in10%glucose solution.

To observe the clinical efficacy of nicorandil for treating the patient with cardiac syndrome X (CSX) and its impact on vascular endothelial function. Methods: A total of 140 CSX patients were randomly divided into 2 groups: Control group, the patients received conventional anti-angina therapy and Nicorandil group, based on conventional anti-angina therapy, the patients received additional oral nicorandil treatment. n=70 in each group. All patients received resting emission computed tomography (ECT) and treadmill exercise ECG stress test (TET). Blood levels of endothelin (ET-1), high-sensitivity C-reactive protein (hs-CRP) and nitric oxide (NO) were examined before and 3 months after treatment. Results: Compared with pre-treatment condition, the attack frequency of angina pectoris and positive rate of ECT were decreased after treatment in both groups, P<0.05; in Nicorandil group, the suspicious positive rate and positive rate of TET were reduced after treatment, P<0.05. Compared with Control group, Nicorandil group had the much lower suspicious positive rate and positive rate of TET after treatment, P<0.05. Blood tests indicated that compared with pre-treatment condition, ET-1 and hs-CRP were decreased, NO was increased after treatment in both groups, all P<0.05; blood levels of ET-1, hs-CRP and NO were different between 2 groups after treatment, all P<0.05. Conclusion: Nicorandil could inhibit inflammatory factors, elevate endothelial function and therefore improve micro vascular angina symptoms, increase exercise tolerance obviously.

Objective: To observe the efficacy of unblocking and regulating the Belt Vessel acupuncture method for patients with abdominal obesity in Germany and its influence on the body fat parameters and glycolipids metabolism. Methods: A total of 82 cases of abdominal obesity in Germany were selected. There were two dropout cases in the treatment process. All of the patients received acupuncture at Daimai (GB26), Daheng (SP15), Tianshu (ST25), Zhongwan (CV12), Shuidao (ST28), Waiguan (TE5), and Zulinqi (GB41). The patients received acupuncture therapy once every 2-3 d, 3 times a week. The treatment duration lasted 4 weeks. Before and after treatment, the body mass (BM), waist circumference (WC), abdominal circumference (AC), hip circumference (HC), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and fasting blood glucose (GLU) were measured respectively, and the body mass index (BMI), waist-to-hip ratio (WHR), waist-to-height ratio (WHtR), and body fat rate (BFR) were calculated. Results: After treatment, the total effective rate was 72.5％; the levels of BM, WC, AC, HC, BMI, WHR, WHtR, BFR, TC, LDL-C, and GLU declined significantly compared with those before treatment (P<0.01), while the TG and HDL-C slightly declined, but the differences were statistically insignificant (P>0.05). Conclusion: The unblocking and regulating the Belt Vessel acupuncture method can significantly reduce body fat parameters such as BM, AC, and BFR, and correct the disorders of glucose and lipid metabolism in patients with abdominal obesity in Germany.

Objective To observe the solo-allied expression of Survivin, Smad4/dpc4 and APC gene, and study the relationship between the three genes and biological behavior of colorectal cancer. Methods The expressions of Survivin, Smad4/dpc4 and APC gene were detected by SP immunohistochem-istry among the cases from 40 eu-intestine ,80 large intestinal adenoma and 80 CRC. Results The positive rate of survivin protein in the eu-intestine, the large intestinal adenoma and CRC was 0,35.0% ,75.0% , respectively. The positive rate of Smad4/dpc4 protein was 100% , 95. 0% , 78. 8% , and the positive rate of APC protein was 100% ,80.0% ,45.0% , respectively. In the eu-intestine, APC + Smad4/dpc4 Z- expressed in 40 cases, with the incidence of 100%. In the large intestinal adenoma, APC + Smad4 expressed in 38 patients, with the incidence of 47. 5% , and the three genes expressed in 29 patients with the incidence of 23. 8%. In the CRC, two genes expressed in 57 patients, with the incidence of 71. 3% , and three genes expressed in 28 patients, with the incidence of 35.0%. Conclusion The detection of survivin was a new indicator in early diagnosis of CRC. It was significant in the diagnosis of CRC that energetic search for the positive rate level realm of survivin. The catastrophe or absence of Smad4/dpc4 gene not only induced the genesis of CRC but also encouraged its growth. Smad4/dpc4 was ant- carcinomatous gene intimately correlated to the CRC. Detection of APC gene had very important significance, it could be helpful to conduct the research of tumorous aetiology, nosogenesis and early diagnosis. The allied detection of Survivin, Smad4/dpc4, APC gene played a very important role in early diagnosing and healing the CRC.

Objective To investigate the ability of proton magnetic resonance spectroscopy (1H-MRS) for differentiating benign from malignant solid adnexal tumors. Methods One-hundred and six patients (114 tumors) with surgically and histologically proven solid adnexal tumors (44 benign, 70 malignant) underwent conventional MR imaging and 1H-MRS. Single-voxel spectroscopy was performed using the point resolved spectroscopy localization technique with a voxel size of 2.0 cm × 2.0 cm × 2.0 cm. Resonance peak integrals of choline (Cho), N-acetyl aspartate (NAA), creatine (Cr), lactate (Lac), and lipid (Lip) were analyzed and the Cho/Cr, NAA/Cr, Lac/Cr and Lip/Cr ratios were recorded and compared between benign and malignant tumors using independent two-sample t test. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic performance of Cho/Cr ratio for differentiating benign from malignant tumors. Results A Cho peak was detected in all 114 tumors, NAA peak in 112 tumors (43 benign and 69 malignant), Lip peak in 70 tumors (21 benign and 49 malignant), and Lac peak in 16 tumors (7 benign and 9 malignant). The Cho/Cr and Lip/Cr ratios were 4.8±2.5, 6.4±4.0 in benign versus 9.6 ± 3.3, 10.5 ± 4.6 in malignant solid adnexal tumors, respectively, with a statistically significant difference (t values were-8.826 and-2.915,P<0.05). The NAA/Cr ratio were 1.4 ± 0.7 in benign versus 1.6 ± 1.0 in
malignant solid adnexal tumors, with no statistically significant difference (t=-1.523,P>0.05). When the Cho/Cr threshold was 7.2 for differentiating between benign and malignant tumors, the sensitivity, specificity, accuracy were 80.0%(56/70),88.6%(39/44) and 83.3%(95/114) respectively. Conclusions The 1H-MRS patterns of benign and malignant solid adnexal tumors differ. The Cho/Cr ratio can help clinicians differentiate benign from malignant tumors.

Objective:To study the astragalus polysaccharides ( APS) effect on immune induced by influenza A virus HA2 eu-karyotic expression vector.Methods: The HA2 encoded by the DNA vaccine vector was efficiently expressed in CHO cells, as determined by reverse transcription polymerase chain reaction ( RT-PCR) and fluorescence analysis.60 rats were divided into six groups randomly,which were immunized with normal saline,pEGFP-N1,pHA2/EGFP+different dose of APS by intramuscular injection.The control sera were collected before injection.After injected the 36th day, sera were collected to analyzing IFN-γ, IL-4 and IgG level.Results:IFN-γ,IL-4 and IgG level of pHA2/EGFP+mAPS group was different from that of pEGFP-N1 group or pHA2/EGFP+lAPS group( P<0.05 ).Conclusion: Middle dose of APS could increase immune induced by influenza A virus HA2 eukaryotic expression vector.

Objective:To study whether high dose astragalus polysaccharides( APS) could affect the expression of pNS1/EGFP that included influenza A virus(IAV) non-structure protein 1(NS1) gene in the tissue.Methods:pNS1/EGFP was constructured with NS1 of IAV.Sixty Kunming mice were divided into three groups randomly.Each group of mice was injected separately with one of the following:pEGFP-N1, pNS1/EGFP and pNS1/EGFP+APS in intraperitoneal injection.The mice were injected by intramuscular injection twice with a 3-week interval between injections.The serum samples and muscle samples were obtained on day 14 and day 28 after the booster injection.Sera IL-4,sera IFN-γ,muscle caspase-3 and muscle NS1 expression were measured in ELISA,Western blot or RT-PCR.Results:There were no significant difference among the different groups in day 14 expect that IFN-γof pNS1/EGFP+APS were lower(P<0.05).IFN-γlevel or IL-4 level of pNS1/EGFP+APS were lower compared with other groups in day 28.caspase-3 of pNS1/EGFP+APS were lower compared with other groups in day 28.Conclusion:APS could increase the expression of pNS1/EGFP by decreasing the inflammation and apoptosis.

BACKGROUND:Hepatocyte transplantation has attracted more and more attention as a therapeutic measure for liver failure and genetic metabolic liver diseases.OBJECTIVE:TO evaluate the efficacy and safety of human hepatocyte transplantation in treating hepatitis B related liver failure in one case by a 2-year follow-up.DESIGN:A case-report of 2-year follow-up.SETTING:No.9 Department of Infectious Diseases,Bioengineering Research Room,the 302 Hospital of Chinese PLA.PARTICI PANT:One inpatient with hepatitis B related liver failure was selected from the 302 Hospital of Chinese PLA.and she was diagnosed according the laboratory tests.The transplanted hepatocytes were originated frOm the healthy liver of a 24-year-old man,who had signed the protocol for liver donation before death.METHODS:The hepatocyte transplantation was completed in the Department of Radiology,the 302 Hospital of Chinese PLA in December 2004.Liver was isolated to obtain human primary hepatocytes, and then cryopreserved.The hepatocytes were transplanted into recipient spleen via femoral vein after resuscitation.The clinical symptoms,changes of blood biochemical indexes,and changes of spleen MRI signals were observed before and after operation.The patient was reexamined every half a year after operation, including liver function, blood coagulation function,B-mode ultrasonography,gastroscopy and MRI,and she was followed up for 2 years. MAIN OUTCOME MEASURES:Liver function,blood coagulation function, imaging indexes, immunological indexes,complication and rejection.RESULTS:①Totally(1-2)×1010 hepatocytes were harvested,and the viability of rewarmed hepatocytes was 60%,and finally 2×109 hepatocytes were transplanted.②Two months later,the clinical symptoms of the recipient were obviously ameliorated,and serum bilirubin and aspartate aminotransferase(AST)were obviously decreased,while prothrombin activity was markedly increased.20 months later,the MRI results showed that there was hepatocyte image in spleen.Two years after operation.the total bilirubin level was 20 μmol/L,direct bilirubin level was 7 μmol/L, alanine aminotransferase was 416.75 nkat/L,AST was 533.44 nkat/L,albumin was 37 g/L,prothrombin activity was 90%,which were all obviously ameliorated as compared with those before operation(474.5 μmol/L,340.3 μmol/L,400.08 nkat/L,1 200.24 nkat/L,38 g/L,25%).The patient left the hospital 2 months later and could do light-burdened job.No complications of hydroperitonia and liver function failure, etc.were observed,and no rejection occurred.Several reexaminations by B-mode ultrasonography all indicated the further aggravations of liver cirrhosis and esophageal varices.She was admitted to hospital for twice because of esophageal varices bleeding,and cured by endoscopic variceal sclerosis therapy.CONCLUSION:Hepatocyte transplantation can ameliorate liver function without rejection,but it cannot relieve portal hypertension.