Objective:To determine aristolochic acid A. in Caulis Aristolochiae Manshuriensis and its preparation. Methods: TLCS Refleciton Saw Tooth Method was used. ? s=323nm. narrow slot: 0.4?0.4nm S X=3.Results: The recovery was 99.86%. RSD was 2.04%. Conclusion: This method is suitable for the content determination of aristolochiae acid A in various traditional Chinese medicine comprising Caulis Aristolochiae Manshuriensis.

Objective To confirm that astrocytes from cerebral cortex of newborn rat can be the target cells of Hantaan virus (HTNV)and Seoul virus (SEOV)infection and to observe changes of astrocytes after different infection time. Methods Astrocytes were prepared from cerebral cortex of newborn rat, and then infected with HTNV and SEOV. The established virus infections were confirmed by detection of virus nucleocapsid protein (NP) and S segment RNA in astrocytes using double-label immunofluoreseence, Western-blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results The astrocytes from cerebral cortex of newborn rat were cultured successfully in vitro, which could be infected by HTNV and SEOV. The number of infected astrocytes and the virus titer in the infected astrocytes kept on increasing along with the extended infection duration. Conclusions Astrocytes from cerebral cortex of newborn rat are the target cells for HTNV and SEOV infection. Then establishment of in vitro cultured astrocytes model for Hantaviruses infection will be helpful for the study on the pathogenesis of hemorrhagic fever with renal syndrome.

Objective To evaluate the efficacy of emergency transcatheter arterial embolization(TAE)for spontaneous rupture of hepatocellular carcinoma(HCC).Methods We retrospectively reviewed 16 cases of spontaneous ruptured HCC which had been treated in our hospital between 1997 and 2005.Emergency TAE was performed in the 16 patients,with 6 cases complicated by hypovolemic.Results Bleeding from the ruptured HCC was stopped at the end of the procedure in 16 patients who had undergone successful embolization.The time from the onset of clinical symprtom to TAE been completed was 1.5-5 hours(mean,3.6 hours),The shortest time was 1.5 hours in 3 patients.Following the achievement of hemostasis by TAE,transcatheter arterial chemoembolization(TACE)2-6 times for each case with average 3.7 and CT guided percutaneous ethanol injection(CT-PEI)5-9 times for every case with arerage 6.8 were performed in 12 cases.3 cases were given up for further treatment.The liver transplantation was carried out in 1 case 3 days after TAE.The average survival time was 14.3 months.Conclusions Emergency TAE is safe and effective for bleeding due to spontaneous rupture in HCC and would achieve better result by adding TACE and CT-PEI.Liver transplantation is one of the options for the patients with poor liver function.The combination of all methods above can prolong patients survival time.(J Intervent Radiol,2006,15:228-231)

Objective: To improve the traditional preparation procedure of effervescent tablets in order to raise stabilization. Methods: The orthogonal design was used for improvement of process. Results: The optimum procedure condition was A 3B 2C 1. That is adding 7.5% citric acid, 11.25% sodium bicarbonate encapsulated by 5% PEG. Conclusion: The new procedure is superior to the traditional procedure, and it is suitable for the requirement of production on a large scale.

Arg-Gly-Asp (RGD) is a unique minimal integrin-binding sequence found within several glycoprotein ligands and also in snake-venom disintegrins. Adinbitor, a protein with 73 amino acid residues including 12 cysteins and an RGD motif, was cloned from Gloydius blomhoffi brevicaudus in my laboratory. As a new member of disintegrin family, adinbitor can inhibit both human platelet aggregation induced by ADP and angiogenensis in vivo and in vitro, the typical characters of disintegrin family. To separate the effect of inhibiting platelet aggregation from that of inhibiting angiogenensis, the motif KGD was introduced into adinbitor cDNA to replace RGD by site-directed and PCR-based mutagenesis. The recombinant protein (recombinant adinbitor (KGD)) was expressed in E. coli BL21 and purified through the His· Bind affinity chromatography. Recombinant adinbitor (KGD) could inhibit ADP-induced platelet aggregation with IC50 value of 85 nmol/L. Considerably, it was a more effective inhibitor on platelet aggregation than recombinant adinbitor (RGD), which has an IC50 of 150 nmol/L. Interestingly, recombinant adinbitor (KGD) has no potency in inhibiting angiogenesis in vivo compared with recombinant adinbitor (RGD). These findings showed that KGD containing adinbitor was more suitable for inhibiting ADP-induced human platelet aggregation as a potential and specific inhibitor of human platelet aggregation, which might have promising therapeutic potential as an antithrombotic agent.

Objective To analyze the complications of total proctocolectomy with ileal pouch-anal anastomosis (TPC-IPAA) in treatment of severe or refractory ulcerative colitis (UC).Methods The retrospective cross-sectional study was conducted.The clinical data of 67 patients with severe or refractory UC who underwent TPC-IPAA in the Xinhua Hospital Affiliated to Shanghai Jiaotong University from February 2008 to October 2015 were collected.All the patients received open and laparoscopy-assisted TPC-IPAA,and two-stage or three-stage surgery was performed according to the patients' conditions.Observation indicators:(1) treatment and follow-up situations,(2) short-term complications after TPC-IPAA,(3) long-term complications after TPC-IPAA.The follow-up using telephone interview and outpatient examination was performed to detect the quality of postoperative life in patients and occurrence of complications up to July 2016.Measurement data with skewed distribution were described as M (range).The comparison of quality of pre-and post-operative life in patients was done using the paired t test.Results (1) Treatment and follow-up situations:all the 67 patients received successful surgery,including 45 with two-stage surgery and 22 with three-stage surgery.Thirty-seven patients underwent open surgery and 30 underwent laparoscopic surgery.The average frequency of postoperative daily defecation,average scores of quality of pre-and post-operative lifes were 5.6,0.47 and 0.67,respectively,with a statistically significant difference between preoperative indicators and postoperative indicators (t =-4.80,P ＜ 0.05).All the 67 patients were followed up for a median time of 4.6 years (range,1.0-8.4 years).(2) Short-term complications after TPC-IPAA:Of 67 patients,10 had short-term complications (some patients with multiple complications).One patient was complicated with presacral abscess secondary to sinus formation and then was cured by topical incision of sinus under colonoscopy.Five patients were complicated with anastomotic site-related complications of TPCIPAA,and 1 with pouch-anal anastomotic fistula combined with presacral abscess underwent drainage with ileostomy and didn't undergo stoma reversion of ileum.Two patients with fistula at the top of pouch were improved by surgery and repair.Of 2 patients with pouch-vagina fistula,1 underwent resection of pouch stump-vagina fistula and then were cured,and the space between pouch stump and posterior fornix was filled with a free greater omentum flap and temporary ileal bypass was simultaneously conducted.The other patient received twice transanal vaginal fistula repairs with advancement flap,and unclosed fistula was confirmed by angiography,with a clinical symptomatic relief.Eight patients with wound infection received actively dressing change after discharge,and no wound dehiscence was occurred.(3) Long-term complications after TPC-IPAA:28 of 67 patients had long-term complications.Twelve patients with intestinal obstruction underwent conservative treatments of gastrointestinal decompression,anti-inflammatory with corticosteroid and anti-infection with antibiotic,without the occurrence of severe intestinal perforation and prognosis needing surgical intervention,including 9 undergoing open TPC-IPAA and 3 undergoing laparoscopic TPC-IPAA.Thirteen patients with postoperative pouchitis received the conservative treatment of mesalazine.Three patients with failed pouch underwent small intestine permanent colostomy,including 2 with secondary pouch Crohn's disease and 1 with severe pouchitis.Conclusions TPC-IPAA is safe and effective in treatment of UC,with a lower incidence of complications.Anastomotic site-related complication of TPC-IPAA is the main short-term complication.And in the long-term complications,pouchitis is the most,followed it is intestine obstruction.

Objective To investigate effects analysis of joint function and MMP-7 and MMP-9 measurement after synovial ar-throscopic debridement for knee synovitis.Methods 72 patients with knee synovitis method based on a random draw were e-qually divided into the treatment group and the control group,the control group were given the traditional synovial planing surgery.The treatment group were given the arthroscopic debridement synovium.Results After treatment,the response rates in the treatment group and the control group were 97.2% and 80.6% respectively.The response rate of the treatment group were significantly better than the control group.The difference was statistically significant (P <0.05).The postopera-tive Lysholm scores were increased significantly,while the pain scores were significantly reduced,and the postoperative Ly-sholm score and pain scores difference compared between the two groups were also statistically significant.The difference was statistically significant (P <0.05).The postoperative synovial tissue MMP-7 and MMP-9 expression rates were in the two groups significantly lower.The difference was statistically significant (P <0.05),while the postoperative MMP-7 and MMP-9 expression rates of the treatment group were ignificantly lower than the control group (P <0.05).Conclusion Syn-ovial arthroscopic debridement for knee synovitis can relieve pain and improve knee function,reduce the expression of MMP-7 and MMP-9,thereby enhance the overall efficacy of the treatment.

Objective To establish reference values for blood pressure in cynomolgus monkeys in different ages.Methods The blood pressures and blood lipids indexes were detected in 521 cynomolgus monkeys using an American BECKMAN-CX4 automatic biochemical analyzer and a wrist electronic blood pressure monitor.Statistical tests were performed to analyze the data.Results Significant differences were found in blood pressure values of cynomolgus monkeys in different ages.Blood pressure values in the elderly group were higher than those of other groups.The morbidity of hypertension in the elderly group was higher than those of the other groups.Body mass index (BMI) in the hypertension group was higher than that of normal group in the same age.The incidence of hypertension in the elderly group with hyperlipemia was higher than that of other groups.Logistic regression analysis showed that age, BMI and hyperlipidemia in the hypertensive group were 1.435, 1.218, and 2.337 times higher than those of the normal group when predicting the risk of hypertension.Conclusions We have initially established reference values of blood pressure in cynomolgus monkeys in different ages.Age, BMI and hyperlipidemia are risk factors of spontaneous hypertension in cynomolgus monkeys, and the measurement of blood pressure may provide a basis for the screening of cynomolgus monkey model of spontaneous hypertensive and related research.

Objective:At present, the postgraduate education of emergency medicine in China mainly relies on the internal medicine system and lacks specialty pertinence. The purpose of this study is to establish a compulsory curriculum system for professional postgraduates of emergency medicine.Methods:Modified Delphi method was used to subscribe questionnaires to experts, and the contents were mainly about medical education management and research of emergency in the affiliated teaching hospitals of Peking University. Microsoft Excel 2016 was used for double entry of questionnaire contents, and SPSS 22.0 software was used for statistics.Results:The panel was consisted of 14 experts in total and two rounds of Delphi questionnaires were completed. The response rate of the two rounds were both 100%. The authority coefficient of experts was > 0.9 and the average score of each evaluation index was 4.07-5.00 points. The coefficient of variation of each index was 0-0.22 and the Kendall coefficient of concordance was 0.07-0.10.Conclusion:As a result, a comprehensive emergency professional postgraduate curriculum system has been established.

Ranpirnase (onconase, ONC) is a new drug, with weak RNase activity and strong cytotoxicity to various tumor cells in vitro and in vivo. This study is to obtain recombination onconase (rONC) with high bioactivity. Based on the codon preference of Pichia pastoris, we designed and synthesized the gene according to cDNA sequences of ONC and the α mating factor's prepeptide. We screened positive clones after transforming the recombination plasmids into P. pastoris X-33, GSS115 and SMD1168. We screened the best combination of seven different vectors and host strains. Moreover, we optimized culture condition in shake flasks and 10 L bioreactor, and purified rONC from the supernatant after inducing it with 0.25% methanol by aqueous two-phase extraction coupling G50 molecular exclusion method. The highest rONC production was 13 mg/L in pPICZα-A/X-33/ONC combination under the condition of pH 5.5 and 23 degrees C in shake flasks for 7 d; and that the highest rONC production was 180 mg/L when the induction is performed in the lower basic salt medium with pH 5.5 in the 10 L bioreactor for 7 d. The yield of rONC is more than 90% at a purity of above 95%. rONC can kill various tumor cells in vitro. The expression and purification of rONC would be useful for further investigation of this new drug.
Antineoplastic Agents ; metabolism ; Bioreactors ; Cell Line, Tumor ; Codon ; DNA, Complementary ; Genetic Vectors ; Humans ; Pichia ; metabolism ; Recombinant Proteins ; biosynthesis ; Ribonucleases ; biosynthesis