Objective: To establish an HPLC method for the determination of mannityl nicotinate and its related substances. Methods:The HPLC method was performed on an Agilent Extend-C18 column (250 mm × 4. 6 mm, 5 μm) with the mobile phase of 0. 01 mol·L-1 ammonium acetate solution-methanol(40∶60). The detection wavelength was 264nm. The column temperature was 30℃. The flow rate was 1. 0 ml·min-1 and the injection volume was 10 μl. Results: The linear range of mannityl nicotinate was 53. 6-1 072. 8 μg·ml-1(r=1. 000 0), and the mean recovery was 98. 9%(RSD=0. 4%,n=9). Mannityl nixotinate and its related substances could be well separated. The limit of quantitation was 0. 06 μg·ml-1 and the limit of detection was 0. 02 μg·ml-1 . Con-clusion:The method is specific, accurate and rapid. It is suitable for the quality control of mannityl nicotinate.

Objective Examinating the efficacy and safety on prophylaxis of deep venous thrombosis(DVT) following arthroplasty using low-molecular-weight heparin (LMWH) in combination with intermittent pneumatic compression(IPC) starting with different time. Methods 156 patients undergoing primary arthroplasty from July 2003 to June 2005 were divided into four groups randomly. In the first group, 38 patients used LMWH preoperatively and IPC starting since anesthesia. In the second group, 42 patients did LMWH postoperatively and IPC starting since anesthesia. In the third group, 36 patients did preoperative LMWH and post-operative IPC. In the fourth group, 40 patients did post-operative LMWH and IPC. All patients were examined by colour duplex ultrasonography, valued the clinical symptom of DVT, recorded the volume of blood loss in perioperation and observed complication of hemorrhage. Results In the first group, two cases of DVT (5.26%) were found and the average perioperative volume of blood loss was (1030.8?282.0) ml. In the second group, two cases of DVT (4.76%) were found and the average volume of blood loss was (900.7?246.9) ml. In the third group, one case (2.78%) of DVT was found and the average volume of blood loss was (1040.3?288.5) ml. In the fourth group, nine cases of DVT (22.5%) were found and the average volume of blood loss was (852.2?295.4) ml. Comparing the incidence of DVT between each group, group 1, 2, 3 were lower than group 4(P﹤0.05). Comparing the volume of blood loss in perioperation, group 2 , 4 were less than group 1,3(P﹤0.05). Conclusion On the basis of this study, it is more effective and safer using LMWH staring with post-operative 10 h in combination with IPC starting with anesthesia on prophylaxis of DVT following arthroplasty.

Objective To compare the recent clinical efficacy between ileal neobladder and orthotopic ileal neobladder.Methods The clinical data of 62 patients with bladder cancer who were performed with cystectomy plus urinary diversion were retrospectively analyzed.Among them,32 patients were performed with orthotopic ileal neobladder(orthotopic ileal neobladder group)and 30 patients were performed with ileal neobladder(ileal neobladder group).The operation time,intraoperative bleeding,intestinal function recovery time,hospital stay,early postoperative complications were compared between two groups.Results All the patients were successful,and there was no operative deaths occurred.There was no significant difference in the postoperative intestinal function recovery time between two groups(P ＞ 0.05).The operation time,intraoperative bleeding,hospital stay in orthotopic ileal neobladder group was higher than that in ileal neobladder group [(463.59 ± 50.24)min vs.(436.07 ± 44.91)min,(1081.16 ± 320.49)ml vs.(867.53 ± 224.61)ml,(46.88 ± 4.67)d vs.(20.37 ± 5.24)d],but the incidence of early postoperative complications in orthotopic ileal neobladder group was lower than that in ileal neobladder group [25.0％(8/32)vs.53.3％(16/30)],and there were significant differences between two groups(P ＜0.05).Conclusions Two kinds of urinary diversion surgical urinary diversion are clinically more mature manner,in clinical practice.Clinicians should be based on the patient's comprehensive situation,combined with the clinical experience to select the most appropriate surgical procedures.

Objective To investigate the influence of epigallocatechin gallate ( EGCG)on reactive oxygen species (ROS) in mouse podocytes exposed to high glucose. Methods Mouse podocytes cultured in high glucose were exposed to different concentrations of EGCG (0.2, 10, 100 μmol/L) or α-tocopherol (0.2 μmol/L) for 6, 12, 24 hours. The viability of podocytes was detected by MTT. The intracellular formation of ROS was detected by confocal microscopy with fluorescent probe CM-H2DCFDA and was measured by fluorescence microscopy. RT-PCR was used to examine the expression of p22phox, p47phox and p67phox mRNA in cultured podocytes exposed to different concentrations of EGCG. Results Intracellular ROS generation was significantly higher in high glucose than that in control conditions (P<0.01). EGCG could significantly inhibit ROS induced by high glucose significantly (P<0.01). EGCG (100 μmol/L) led to an inhibition of the increased production of NADPH oxidase components of p22phox and p67phox mRNA in high glucose (P<0.05). The expression of p47phox mRNA in high glucose was inhibited by EGCG(0.2 μmol/L) and ot-tocopherol(0.2 μmol/L) (P<0.05). Conclusion EGCG can protect cultured mouse podocytes from injury of high glucose by inhibiting ROS formation.

Objective To study the effects of the interaction of advanced glycation end products (AGEs) and the receptor of AGEs (RAGE) on apoptosis of mice podocytes. Methods Podocytes were exposed to soluble AGEs such as bovine serum albumin (BSA), carboxymethyl-lysin (CML)-BSA, AGE-BSA and matrix-bound AGEs (AGE-modified collagen Ⅳ ), and to different concentrations of AGE, such as 10 mg/L, 50 mg/L, 100 mg/L. Apoptosis was assessed by TUNEL staining. Fluorescence-activated cell sorting (FACS) was used for the quantification of apoptotic andnecrotic podocytes after Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) labeling. Apoptosis was described as the ratio of apoptotic cells to the total number cells under the high-power field, siRNA was transfected into podocytes through combining Dharmacon on Targetplus SMART pool siRNA reagents and Amaxa RNAi nucleofection kit. Results The apoptosis rate was higher in podoeytes exposed to either CML-BSA or AGE-BSA than that exposed to BSA. There was a two- to three-fold increase in apoptosis when podocytes were cultured in AGE-modified collagen Ⅳ as compared with native collagen Ⅳ. The apoptotic response of podocytes to AGE-BSA exposure occurred in a dose-dependent manner. Podocyte necrosis occurred only at the highest concentration of AGE-BSA(100 mg/L). AGE-BSA failed to induce apoptosis in podocytes transfected with RAGE siRNA. RAGE-specific gene knockdown did not significantly reduce the apoptosis of podocytes cultured in AGE-modified collagen IV. Conclusions The AGE-RAGE interaction plays a major role in the apoptosis of podocytes triggered by soluble AGEs, but not by matrix-bound AGEs. Reduction of AGE burden and RAGE expression may be important therapeutic approaches to prevent the progression of kidney disease.

Objective To discuss perioperative features, operative approach and surgical effects of spinal tuberculosis in older patients. Methods Retrospective analysis was conducted to analyze the clinical data of 45 patients with spinal tuberculosis ranging in age from 61 to 78 years (average, 67 years). The lesion was located in thoracic vertebrae in 9 patients, thoracolumbar vertebrae in 14, lumbar in 16, and lumbosacral in 6. Preoperative Frankel grades were B in 7 cases, C in 21, D in 11 and E in 6. Among them, 21 had cardio-cerebrovascular disease, 10 had diabetes mellitus. With preoperative medicine and chemotherapy for 2-3 weeks, all patients were treated surgically. The surgical procedures included: 1) Posterior focus debridement, bone grafting and one-stage posterior transpedicular screw system fixation in thoracic vertebrae (T2-T7). 2) Anterior debridement, bone grafting and one-stage posterior transpedicular instrumental fixation in lumbosacral vertebrae (L5-S1). 3) Anterior focus elimination, bone grafting and one-stage anterior plate fixation in the other vertebrae. Results Forty-five patients were followed up for 24 to 40 months, with the average of 28.5 months. No severe complication occurred during and after operation. Forty-four cases had recovered and 1 recurred. Spinal fusion occurred 12-18 months after operation. Frankel neurological grades improved significantly. Conclusion With the effective management of comorbidities in perioperative period,the elderly could tolerate surgical treatment. The appropriate approaches, thorough debridement and reasonable bone grafting with internal fixation are key to therapeutic success.

Objective To investigate the long-term efficacy of transendocardial injection of vascular endothelial growth factor (VEGF) into ischemic porcine myocardium.Methods Thirty pigs were divided into two groups:control group and treatment group (15 each).A angioplasty balloon was used to occlude the left anterior descending coronary arteries of the animals to reproduce a porcine myocardial ischemic model.Blank plasmid (control group) or constructed pIRES2-EGFP-hVEGF165 eukaryotic expression plasmid (treatment group) was directly injected into the ischemic myocardium through an transendocardial injection catheter of NOGA system,respectively.Before and one year after injection,left ventricular electromechanical mapping (LVEMM) was used to detect local linear shortening (LLS) and unipolar potential (UpV);M-mode local wall motion,cycle variation of integrated backscatter (CVIB) and left ventricular ejection fraction (LVEF) were detected by ultrasonagraphy.One year later,the animals were sacrificed for the observation of the capillary formation in myocardial tissue with immunohistochemical staining.Results One year after injection,as compared with that of control group,local linear shortening (LLS) showed a significant increase (P0.05),and ultrasonagraphy showed better heart function in the treatment group (P

Objective· To investigate whether vitamin E could reverse the disruptive effects of bisphenol A (BPA) on steroidogenesis and to explore the optimal vitamin E concentration.Methods· Rat primary granulosa cells were extracted and exposed to BPA (0,0.01,0.1,1,10,100 μmol/L).After 48 h of incubation,culture medium was collected and estradiol (E2) and progesterone (P4) were measured using ELISA kits.Then,granulosa cells were incubated with 5 μmol/L(average concentration in follicular fluid) or 25 μmol/L (high concentration in follicular fluid) vitamin E (α-tocopherol) or vitamin E (5 μmol/Lor 25 μmol/L) plus BPA (10 μmol/L or 100 μmol/L) for 48h,E2 and P4 were measured.Results· BPA at 10 μmol/L and 100 μmol/L suppressed E2 and P4 production in a dose-dependent manner (P＜0.05).Vitamin E at 25 μmol/L significantly increased E2 and P4 levels by (44.89±15.18) ％ and (43.33±8.82) ％(P＜0.05),respectively.Coincubation of the granulosa cells with BPA and vitamin E (5 μmol/L or 25 μmol/L) restored the productions of E2 and P4,which were not significantly different from the control group (P＞0.05).Conclusion· Vitamin E (5 μmol/L/25 μmol/L) could reverse BPA-induced reduction in steroid hormone production in rat ovary granulosa cells,and the antagonistic effect of 25 μmol/L vitamin E was more obvious than that of 5 μmol/L vitamin E.

Objective To investigate the efficiency of ring butterfly sewing in lower uterine segment aided with tourniquet in treating pernicious placenta previa combined with placenta percreta. Methods Twelve pregnant women diagnosed with pernicious placenta previa combined with placenta percreta by prenatal ultrasound in Peking University First Hospital from April 1, 2012 to November 30, 2014, were enrolled. All of them received elective cesarean delivery using ring butterfly sewing in lower uterine segment aided with tourniquet. The efficiency of this novel technique, in term of blood loss and vital signs during operation, blood transfusion, and postoperative recovery, was analyzed. Results Twelve cases were diagnosed with placenta percreta during operation, with placenta invading the perimetrium in seven cases and urinary bladder being involved in one case. Ring butterfly sewing in lower uterine segment aided with tourniquet during elective cesarean delivery was all successful in the 12 cases. The median amount of blood loss was 1 000(400-2 000) ml during operation and the amount of blood loss was lower than 1 000 ml in three cases. Two cases received no blood transfusion, and the median blood transfusion in the other 10 cases were 400(400-1 200) ml. The mean operation time was (75±22) min (43-131) min. Eleven cases had normal lochia and temperature after operation and the other one had placenta invading urinary bladder and partial placenta in situ, and developed fever at day 3 after operation, but recovered after antibiotic treatment, and no placenta tissue was seen by ultrasound at day 59 after operation. Conclusions The ring butterfly sewing in lower uterine segment aided with tourniquet is highly effective for the pernicious placenta previa combined with placenta percreta, and it is a simple, fast and fertility sparing technique with low blood loss and without usage of special equipment.

Objective To observe the effects of Huazhuo Granule on expression of phosphatidylinositol-3-kinase (PI-3K) and glucose transporter 4 (GLUT4) in skeletal muscle type 2 diabetes (T2DM) rats;To discuss its effects on activating signal path of PI-3K and GLUT4. Methods High-fat diet combined with low-dose streptozotocin intraperitoneal injection was used to establish T2DM rat models. The rats were randomly divided into blank group, Huazhuo Granule group, rosiglitazone group, and model group. Each treatment group was given relevant medicine for gavage. Model group and blank control group were given normal saline for gavage. The general state, weight, and food ration of rats were observed. After ten-week medicine intervention, 10%chloral hydrate was used for anesthesia to detect the levels of FBG, OGTT (2 h PG), and FINS. RT-PCR was used to detect the expressions of PI-3K and GLUT4 mRNA of rat skeletal muscle in all groups. Results Compared with model group, the levels of FBG, 2 h PG, and FINS decreased (P<0.05);the expressions of PI-3K and GLUT4 mRNA of rat skeletal muscle increased significantly (P<0.05). Conclusion Huazhuo Granule has the effects on increasing insulin sensitivity, which is related to activating insulin signal path of PI-3K and GLUT4 of skeletal muscle tissues.