OBJECTIVE: To establish an HPLC method for the content determination of prostaglandin A1 and prostaglandin B1 in Alprostadil for injection.METHODS: The determination was performed on Alltech Alltima C18 column with mobile phase consisted of phosphate puffer(pH=6.3)-acetonitrile-methanol(70 ∶ 25 ∶ 5) at a flow rate of 1.5 mL? min-1.The detection wavelength was set at 196 nm.The column temperature was set at room temperature and the injection volume was 20 ?L.RESULTS: The prostaglandin A1 and prostaglandin B1 were well separated from main component and other impurities.The linear range of prostaglandin A1 and prostaglandin B1 were 0.175～19.00 ?g?mL-1(r=0.999 7) and 0.23～19.90 ?g?mL-1(r=0.999 2).The contents of prostaglandin A1 in 3 batches of samples were 4.7%,4.9% and 4.3%,and the contents of prostaglandin B1 in 3 batches of samples were 0.6%,0.8% and 0.5% respectively.CONCLUSIONS: This method is proved to be simple,specific and suitable for the content determination of related substances in Alprostadil for injection.

BACKGROUND:To build up an effective method of isolating and culturing granule cels is a pivotal step to enhance fertilization-embryo transfer rate. Current studies mainly focus on the isolation methods of human ovarian granulosa cels rather than cel counting, purity and subsequent growth. OBJECTIVE: To establish the effective methods of isolating, purifying and culturing human ovarian granulosa cels in vitro. METHODS: Folicular fluid was harvested from women undergoing fertilization-embryo transfer procedures. Human ovarian granulosa cels were obtained from the folicular fluid by lysis treatment, precipitation method or density gradient centrifugation. Granulosa cel mucus masses were digested with type I colagen enzyme or hyaluronidase and then cultured in the culture medium with or without autologous folicular fluid. RESULTS AND CONCLUSION: Lysis treatment yielded the largest amount of granulosa cels compared to the precipitation method and density gradient centrifugation (P > 0.05,P < 0.05, respectively). Cels prepared by the three methods showed the same cel viability. After 24 hours of culture, the precipitation method obtained the largest amount of adherent granulosa cels (P < 0.05); and the density gradient centrifugation obtained the least amount of cels (P < 0.05). Compared with type I colagen enzyme, hyaluronidase took less time to digest the cels thoroughly. Autologous folicular fluid could promote the growth and survival of granulosa cels. These findings indicate that the precipitation method, though time-consuming, can obtain the highest cel viability and harvested the largest amount of granulosa cels after culture; hyaluronidase is more suitable for digesting granulosa cel mucus mass than type I colagen enzyme; autologous folicular fluid added into the culture medium is more conducive to granulosa cel growth.

Objective To investigate the effect of continuous positive airway pressure(CPAP)during one lung ventilation on pulmonary function in patients undergoing video-assisted thoracoscopic repair of atrial septal defect.Methods Twenty ASA Ⅱ patients of both sexes,aged 16-30 yr,weighing 41-64 kg,scheduled for video-assisted thoracoscopic repair of atrial septal defect,were randomly divided into 2 groups(n = 10 each): control group and CPAP group.One lung ventilation(VT 8 ml/kg,RR 12-16 bpm,I:E 1:2,PETCO2 35-40 mm Hg)was perform in both groups.CPAP(6 cm H2O)was perform during one lung ventilation in group CPAP.The oxygenation index,pulmonary compliance and airway pressure were recorded during operation.The cardiovascular events,hyoxemia and extubation time were recorded.Results The oxygenation index and pulmonary compliance were significantly higher,extubation time was shorter,and the incidence of hyoxemia was lower in CPAP group than in control group(P ＜ 0.01).The airway pressure was in the normal range in both groups.No cardiovascular events was found in both groups.Conclusion CPAP(6cm H2O)during one lung ventilation can improve the pulmonary function in patients undergoing video-assisted thoracoscopic repair of atrial septal defect.

Objective:To investigatethe transdermal delivery characteristics of methylphenidate hydrochloride (MPH) in vitro. Methods: Characteristics of MPH crossing nude rats skin were studied with Franz diffusion cells. A high performance liquid chromatographic (HPLC) method was established to determine the concentration of MPH crossed the skin. The permeability coefficient (P), steady state flux (J) and lag time(LT) for MPH through the skin of nude rats treated with various enhancers were compared with those of control. Results: The permeability coefficient increased with the increase of MPH concentration. The penetration of MPH through nude rats skin was obviously enhanced by 8%Azone and 5%propylene glycol (P

Objective: To investigate arsenic trioxide (As 2O 3) -target interactions at the level of nuclear matrix (NM) in chronic myelogenous leukemia cell line K562 by proteomics. Methods: DNA fragmentation analysis was used for As 2O 3 induced apoptosis of K562 cells. The nuclear matrix proteins were analyzed by high-resolution two-dimensional gel electrophoresis and computer-assisted image analysis. Results: While more than 200 protein spots were shared among the nuclear matrices, about 18 distinct spots were found characteristic of As 2O 3 treated cells. Onset of mass mange apoptosis, and the profiling of nuclear matrix proteins had been alternated and it was a more sensitive indicator than nucleosomal DNA fragmentation against As 2O 3 treatment. Conclusion: As 2O 3 induced apoptosis in K562 cells in a dose-time-dependent manner. As 2O 3 might be clinically useful in treatment of chronic myelogenous leukemia and the changes of nuclear matrix proteins in the treated cells can be used as a useful indicator for the treatment.

Objective To explore the early diagnostic value of Golgi membrane protein 73 (GP73),alpha-fetoprotein (AFP) and alpha-fetoprotein-L3 (AFP-L3) in patients with high risk of primary hepatic carcinoma (PHC).Methods Sixty-four cases of PHC were selected as the PHC group,60 cases of liver cirrhosis(LC) as the LC group,53 cases of hepatitis as the hepatitis group and 51 healthy checked-up people as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum level of GP73 in all the cases.AFP-L3 was isolated by using affinity micro centrifugal column,AFP and AFP-L3 were detected with chemiluminescent immunoassay and then the proportion of AFP-L3 was calculated.Results The positive rate of serum GP73,AFP and AFP-L3 in PHC group was significantly higher than that in LC group and hepatitis group [78.1％ (50/64)vs.25.0％ (15/60),17.0％ (9/53);48.4％ (31/64) vs.31.7％ (19/60),22.6％(12/53) ;53.1％(34/64) vs.30.0％(18/60),20.8％(11/53)] (P ＜ 0.05),In control group,GP73,AFP,AFP-L3 was no positive.The levels of GP73,AFP and AFP-L3 in PHC group were significantly higher than those in LC group,hepatitis group and control group [(245.69 ± 89.18)μ g/L vs.(116.37 ±38.52),(97.29 ± 24.58),(23.48 ±9.12) μ g/L; (403.27 ± 128.46) μg/L vs.(75.62 ± 19.35),(66.49 ± 15.14),(3.46 ± 1.02) μg/L; (15.64 ±3.19)％ vs.(5.24 ± 1.15),(4.21 ± 0.96),(2.95 ±0.73)％] (P ＜0.05).The levels of GP73,AFP in LC group and hepatitis group were significantly higher than those in control group (P ＜ 0.05).The levels of GP73,AFP and AFP-L3 had no statistically significant difference between LC group and hepatitis group (P ＞ 0.05).Sensitivity and accuracy of three combined detection for PHC was 96.9％(62/64),91.7％(209/228),significantly higher than that of AFP,AFP-L3 single detection (P ＜ 0.05).GP73 single detection and any two combined detection was no significant difference in sensitivity and accuracy,compared with three combined detection (P ＞ 0.05).The levels of GP73 in PHC patients with different age,gender,serum level of AFP,TNM stage and tumor diameter had no statistically significant difference (P ＞ 0.05).The levels of GP73 in PHC patients with positive HBsAg,extrahepatic metastases and LC had significant difference (P ＜ 0.05).Conclusions The diagnosis value of GP73 is evidently higher than AFP and AFP-L3 for PHC,and combined determination is superior to single marker.Combined determination enhances the degree of precision in populations with high risk of PHC diagnosis.

Humans ; Male ; Splenectomy ; Stem Cell Transplantation ; adverse effects ; Thrombocytopenia ; etiology ; surgery ; Thrombotic Microangiopathies ; complications ; surgery ; Young Adult

Objective To establish a new model of acute hepatic failure in canines for bioartificial liver treatment research . Methods Acute hepatic failure was induced by end to side portocaval shunt combined with common bile duct ligation and transection. The survival rate, liver function, ammonia, Fischer's ratio (BCAA/AAA) and pathological change of injury liver were investigated. Results The ammonia, ALT, AKP, total bilirubin and PT were elevated dramatically and Fischer's ratio declined significantly 14 days after the operation. The survival rate of the models decreased with time 14 days after the operation. Pathological examination of liver tissues revealed evidence of cholestasis, inflammation with accumulation of neutrophils, degeneration and necrosis of hepatocytes.Conclusions End to side portocaval shunt combined with common bile duct ligation and transection can produce a satisfactory acute hepatic failure model in canines, which could be used in the study of bioartificial liver.

AIM: To evaluate the protective effects of mycophenolate mofetil on the kidneys of type 2 diabetic rats and discover their mechanisms. METHODS: Wistar rats were divided into three groups, such as normal control rats, diabetic rats, and diabetic rats in the treatment with mycophenolate mofetil (MMF, 15 mg?kg -1 ?d -1 ). Thirteen weeks later, urinary albumin excretory rate (UAE), creatine clearance (Ccr), blood glucose, blood insulin and blood lipid were measured, and kidney pathology was observed. Inmmunohistochemistry was used to analyze the expression of CTGF, ColI and ColⅢ. RESULTS: Mycophenolate mofetil decreased UAE, Ccr and reduced glomerular volume. The expression of CTGF and deposition of ECM decreased after diabetic rats received mycophenolate mofetil. CONCLUSION: Mycophenolate mofetil can protect the kidney of diabetic rats. Its mechanism may be related to the decrease of CTGF expression and extracellular matrix deposition in renal tissue.

The structural protein VP6 of rotavirus form the middle layer of the triple-layered viral capsid, playing a key role in the organization of the virion. The gene of structural protein 6 of rotavirus strain TB-Chen isolated from a clinic sample was amplified using PCR from the reverse transcription product of RV genome RNA, using pET as expression vector, a recombinant plasmid pET-VP6 containing coding sequence of VP6 protein was constructed. The results showed that the VP6 was highly efficiently expressed in E. coli BL21(DE3) cells which were transformed with the recombinant plasmid pET-VP6.The expressed VP6 protein possessed 27.4% of total cells protein, with an approximately 45kDa of molecular weight, and could be recognized by guinea pig anti-SA11 antibody on Western blot. The results obtained provide important basis for further study on structure and function of the VP6 protein.