Objective To establish an high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method for determination of glucosamine hydrochloride and its related substances in glucosamine and indometacin enteric-coated capsules.Methods Separation was carried out on a Inertsil C8 Column (250 mm ×4.6 mm,5μm) with a mobile phase composed of acetonitrile-water(70∶30) at a flow rate of 1.0 mL/min and the column temperature was 25℃.Results The linear ranges of calibration curve was 25 -312.5 μg/mL(r=0.9991, n=6).The average recovery was 98.0%, and the relative standard deviation was 1.3%( n =5 ) .The glucosamine hydrochloride was stable with acid destruction, but unstable with alkali, heat and strong light destruction.Conclusion The method is simple, accurate and reproducible for determination of glucosamine hydrochloride and related substances.

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A total of 206 patients of type 2 diabetes mellitus (T2DM) were assigned into the trial group (n =156) and the control group (n =50) according to whether real object-based diabetic diet was provided.Traditional nutritional education for both groups during hospitalization.The patients in the trial group received a real object-based diabetic diet guided by a nutritionist who adjusted feeding regimens and skills before and after operation.The control group had a free access to a normal diet.The patient understanding of knowledge,attitude and practice (KAP) was examined by nutrition surveys.And their postoperative nutrition states and complications were assessed by questionnaires during a 6-month follow-up.The total average score of cognitive divided was 93.4 ± 7.5,the average score of diet therapy knowledge 13.8 ± 1.6 and the average scores of attitude and practice 9.6 ± 1.9 and 32.2 ±3.9.Significant differences existed between two groups (P ＜ 0.05).The incidence rates of hypoglycemia,hyperglycemia and gastrointestinal obstruction were lower than the control group (P ＜ 0.05,P ＜ 0.01).It suggested that real object-based diabetic diet guidance plus traditional nutrition education could improve the KAP level and may lower the incidence of postoperative complications in T2DM patients with gastric bypass.

Objective To observe the effect of small interfering RNA(siRNA) targeting Survivin gene on survivin expression,proliferation and apoptosis of hepatocellular carcinoma cell line MHCC-97H.Methods Survivin sequence specific siRNA was designed and synthesized.siRNA/liposome complex was transfected into hepatocellular carcinoma cell line MHCC-97H.The MHCC-97H cells were divided into Survivin siRNA group(Si-survivin),negative control siRNA group(NC group)and blank group (normal control group).Survivin mRNA and protein expressions were detected by reverse transcription-PCR and Western blot,respectively.The proliferation of MHCC-97H was measured by methythiazolydiphenyl-tetrazolium bromide assay.The Annexin V/PI double labeled flow cytometry was employed to measure the apoptosis at 24 h after transfection in different concentrations of Survivin siRNA(12.5,25.0 and 50.0 nmoL/L,respectively).Results After 48 h of transfection,the Survivin mRNA levels were 0.55 ± 0.16 (Si-survivin group),0.85 ± 0.28 (NC group) and 0.93 ± 0.40 (normal control group),respectively,which were significantly different among 3 groups (F =414,P ＜ 0.01).The level of Survivin mRNA was the lowest in Si-survivin group,which was statistically different with NC group and normal control group (t =-20.56,-28.37,all P ＜ 0.001).The levels of Survivin protein expression in 3 groups were 0.602 ± 0.005 (Si-survivin group),0.835 ± 0.007 (NC group) and 0.993 ± 0.003 (normal control group) at 48 h after transfection,which were statistically different among 3 groups (F =238,P ＜0.01).The lowest level of protein expression was in Si-survivin group,which was statistically different with NC group and normal control group (t =-40.17,-66.03,all P ＜ 0.001).After 72 h and 96 h of transfection,the inhibitory rate of cell growth was significantly higher in Si-survivin group [(19.5 ± 3.6)％,(12.0 ± 0.9)％] compared with that in NC group [(3.6 ± 0.9) ％,(-1.3 ± 6.1) ％] (t--36.18,42.53,all P ＜ 0.05).The apoptosis rates in 12.5,25.0,50.0 nmol/L Survivin siRNA were (22.64 ± 2.54) ％,(35.37 ± 3.28) ％ and (53.28 ± 4.35) ％,respectively.However,in NC group and normal control group,the apoptosis rates were (8.77 ± 1.25) ％ and (9.72 ± 1.37) ％.The rates were statistically different among those 5 groups(F =35.93,P ＜0.01).And in the apoptosis rates of siRNA groups in different concentratiom were statistically different when compared between each two groups (t =-29.73,-38.57,all P ＜ 0.001).Conclusion Survivin specific siRNA can inhibit the proliferation and induce the apoptosis by blocking Survivin gene expression in hepatocellular carcinoma cell line MHCC-97H.

The widespread availability of multi-detector spiral CT and its post processing techniques,and MRI,provide us with important tools to carry out anatomical studies on pancreaticobiliary maljunction (PBM).Determining the relationship between PBM and biliary diseases contributes not only to a better understanding of pathogenesis and pathological process of biliary diseases,but it also provides evidence for prophylactic treatment.The present study reviewed the definition,classification,pathological anatomy of PBM,relationship between PBM and pancreaticobiliary diseases,and clinical treatments of PBM.

Objective:To investigate the effects of mucosal strengthening treatment on preventing recurrence of closedvarices. Methods: In 264 patients with esophageal varices obliteration by banding ligation and sclerotherapy, of 136 patientswere gone on to treat with strengthening mucosa, and 128 patients without strengthening treatment. All patients had beenobserved out of hospital for 1-13 years. Results: There were no recurrence and rebleeding of esophageal varices in the groupwith strengthening treament. But in the group without strengthening treatment, 25 patients (19. 5%) had isolated or strip-like varices with red colour signI small veins grew thick and part of mucosa appeared clear hyperaemia in 21 patients(16.4% ); 6 patients (4.7% ) rebleeded. There were significant differences between two groups (P

Preoperative diagnosis of biliary obstruction mainly depends on imaging examination.Percutaneous transhepatic cholangiography (PTC) is a common method in detecting biliary obstruction.PTC combined with computed tomography (CT) could enhance the diagnostic rate.From April 2009 to April 2011,8 patients with biliary obstruction were admitted to the Shenzhen Hospital of Peking University.Contrast solution (30 ml of iodine solution at a concentration of 1.5％ -2.0％)was injected through a PTC tube,and then CT scan was performed.An iohexol contrast solution at a concentration of 300 mgl/ml was injected at a dosage of 1.5 ml/kg and at 3-5 ml/s,then the arterial phase,venous phase and delayed phase were scanned.The original data were uploaded to Vitrea 2 workstation for multiplane reconstruction,maximum intensity projection and volume rendering.The procedure was successfully performed on all patients,and the position of the biliary obstruction was identified in 7 patients.Five patients were diagnosed as with hilar cholangiocarcinoma,1 with sclerosing cholangitis and 2 with adenoma of the distal common bile duct.The patients' symptoms were alleviated after percutaneous transhepatic biliary drainage.

Objective To explore the proliferation characteristics of primary small intestinal epithelial cells of tree shrews and the characteristics of human rotavirus(RV) G1P[8] infection to these cells,and establish a model of tree shrew primary small intestinal epithelial cells infected with human rotavirus G1P[8].Methods The primary small intestinal epithelial cells were obtained by collagenase Ⅺ and dispase I digestion from tree shrew.After purification and identification,the obtained primary small intestinal epithelial cells were infected with RV.Then,culture supernatants of infected cells were collected every 12 hours after infection.Viral titer and viral load were subsequently determined.Western blot and indirect immunofluorescence observation were used to detect the expression of RV protein VP6 in the primary cells.The infectivity of RV to the tree shrew primary cells was finally evaluated.Results After purification and identification of primary epithelial cells from the tree shrew,high purity above 90% primary tree shrew small intestinal epithelial cells was obtained.These primary small intestinal epithelial cells could be infected with RV virus by comparing the virus infectivity to primary renal cells,HCT116 cells and MA104 cells.The virus titer reached to 2.0×105TCID 50/mL at 72 h after infection.Using Western blot and indirect immunofluorescence observation,the specific viral protein of VP6 was determined to be expressed in the tree shrew primary small intestinal epithelial cells,and were located in the cytoplasm from days 1 to 5.Conclusions The separation,purification and cultivation methods of tree shrew primary small intestinal epithelial cells are successful,and the tree shrew model of RV-infected the tree shrew primary small intestinal epithelial cells is successfully established.

OBJECTIVE: To explore the genetic basis for an infant with congenital generalized lipodystrophy. METHODS: Peripheral blood samples of the child and her parents were collected for the extraction of genomic DNA. All exons and flanking sequences of the AGPAT2 gene were subjected to Sanger sequencing. RESULTS: The child was found to harbor compound heterozygous c.792_805delGGAGAACGGGGCCA (p.Gln264Hisfs*208) and c.335C>T (p.P112L) variants in exons 6 and 3 of the AGPAT2 gene, which were respectively inherited from her mother and father. c.792_805delGGAGAACGGGGCCA (p.Gln264Hisfs*208) was previously unreported, while c.335C>T (p.P112L) was known to be pathogenic. CONCLUSION The compound heterozygous variants of the AGPAT2 gene probably underlie the disease in this child.

Objective : To investigate the application of indocyanine green in near-infrared fluorescence imaging to determine the scope of necrotic bone resection in osteoradionecrosis of the jaw and to provide a reference for clinicians Methods : Eight patients with osteoradionecrosis of the jaws were enrolled. Indocyanine green was intravenously injected through the elbow vein 10 minutes before osteotomy. After conservative resection of necrotic bone lesions based on imaging results, the scope of potential dead bone resection in the area of low fluorescence intensity was gradually expanded at an initial distance of 0.3 cm. Near-infrared fluorescence imaging and fluorescence intensity determination of bone cross-section were performed before and after extended resection. Statistical differences were analyzed. All patients with osteonecrosis underwent regular follow-up to evaluate the postoperative efficacy Results: Indocyanine green was injected into all 8 patients with osteoradionecrosis for near-infrared fluorescence imaging and the scans were clear; the fluorescence intensity of fresh bone wounds with an expanded mandibular resection range of (0.95 ± 0.14) cm was (226.2 ± 15.8) au, which was higher than that based on intraoperative macroscopic observation and radiological results (108.8 ± 3.4) au, (t = 20.718, P<0.001). The postoperative follow-up improvement rate of 8 patients was 87.5%. Conclusion Near-infrared fluorescence imaging with indocyanine green can assist in the successful removal of necrotic bone until fresh bleeding of the jaw wound occurs, which has important clinical value in defining the resection range of osteoradionecrosis of the jaw.