AIM:To develop a new method to determine cucurbitacin B in Pedicellus Melo by capillary zone(electrophoresis(CZE).) METHODS:All the samples were analysed on a fused silica capillary(75 cm?75 ?m I.D.) by using 50 mmol/L sodium borate solution(containing 5% acetonitrile) as the background electrolyte with the running voltage at 12.0 kV and detection wavelength of 265 nm.And clorprenaline hydrochloride was applied as the internal standard. RESULTS:The results showed a good linear correlation between relative peak area of cucurbitacin B(y) and its concentration(x) over the range of 0.15～1.2 mg/mL.Regression equation was y=(0.003 4x)+0.058,r=0.999 7,and the average recovery for baicalin was 100.2% with the RSD at(2.11%). CONCLUSION:The method is rapid,simple,reproducible and produces low pollution.It serves as a novel way to determine cucurbitacin B.

Objective　To prevent the after cataract induced by lens epithelial cells proliferation from postoperative cataract, monoclonal antibody (McAb) specific for rabbit lens epithelial cell is made, it will be the carrier further to be conjugated with cytotoxin. The conjugations will inhibit lens epithelial cells growth and not damage the other tissues of eye. Thereby McAb is the experimental bases of preventing after cataract.Methods　BALB/c mice were immunized by mixture of rabbit lens epithelial cells and Freund's adjuant. The immunized mouse spleen cells were fused with parental mouse myeloma cells (BALB/c　SP2/0) using polyethylene glycol (PEG-4000). The fused cells were selectively cultured by hypoxanthine, aminopterin and thymidine (HAT) culture medium. The specificities of the supernatant from hybridomas were tested by indirect immunofluorescence and immunohistochemistry SP (Streptavidin peroxdase conjugated method). The positive hybridomas were further cloned three times by methylcellulose culture medium to ensure monoclonality. At last, the consensual reaction of McAb was tested on human eye tissues.Results　Hybridomas were produced by fusion of spleen cells of immunized mice and mouse myoloma cells (SP2/0) with PEG-4000, and grown selectively in medium containing HAT after 16 days. Antibodies of the supernatant from hybridomas were tested on frozen sections of rabbit lens epithelial cell by indirect immunofluorescence. Only a positive clone secreted McAb against antigen of rabbit lens epithelial cell. The specificity of McAb was tested on paraffin sections of whole rabbit eye and whole human eye by immunohistochemistry SP. The results indicated that McAb was only positive to rabbit lens epithelial cell membrane and it was negative to the other tissues of rabbit or whole human eye tissues.Conclusions　McAb specific for rabbit lens epithelial cell was manufactured successfully. The specificity of McAb was strong. There was no consensual reaction on human eye tissues. It might be the experimental bases of further targeting chemotherapy on after cataract.

Objective To evaluate the efficacy and safety of sirolimus and tacrolimus after renal transplantation.Method PubMed,Web of knowledge,Medline and the Cochrane controlled trials register,Chinese Biomedicaldatabase,and Vip database were searched with the terms and Boolean operators as (kidney transplantation OR renal transplantation) AND (sirolimus OR rapamycin OR rapamune) AND (tacrolimus OR FK506 OR prograf).Results retrieved were updated on November,2015.Data were extracted for patient and graft mortality,acute rejection (AR),wound complications,infection,GFR,withdrawl.Professional meta analysis software RevMan 5.3 was employed.Result Altogether,1810 patients from 10 randomized controlled trials (RCTs) were included.Patients in the sirolimus group showed a decreased rate of graft mortality and infection (RR =0.63,95％ CI,0.45-0.89,P=0.009;RR=4.42,95％ CI,1.73-11.31,P=0.002).Patients in the sirolimus group showed an increased rate of AR,wound complications,GFR,withdrawl (SMD=-0.52,95％ CI,-0.73-0.31,P＜0.000 01;RR=0.54,95％ CI,0.40-0.73,P＜0.000 1;RR=0.17,95％ CI,0.11-0.25,P＜0.000 01;RR =0.44,95％ CI,0.37-0.51,P＜0.000 01).The patient mortality was insignificantly different between two groups.Conclusion This meta-analysis concluded that sirolimus showed advantage over tacrolimus about safety when used early after renal transplantation.The options of immunosuppressive regimens after kidney transplantation should be based on the specific condition.To obtain more reliable and accurate clinical data,the RCTs with more rational design,higher methodological quality,larger sample size,including domestic patients,longer follow-up are still needed.

Objective: To study effect of small ubiquitin related modifier protein 1 (SUMO1) in inflammatory reactions mediated by tumor necrosis factor (TNF)-α and nuclear factor (NF)-κB in myocardial damage of rats with type 2 diabetes mellitus (T2DM). Methods: A total of 20 Goto-Kakizaki (GK) rats with spontaneous diabetes mellitus (DM) were randomly divided into group DM1 (pure DM group, n=10) and group DM2 (DM+high-fat diet group, n=10), and another 10 normal Wistar rats were regard as healthy control group. Expressions of SUMO1, TNF-α and NF-κB were measured by immunohistochemical method. Results: 1. Levels of blood glucose and TG in group DM1 and group DM2 were significantly higher than those of healthy control group, and those of DM2 group were higher than of DM1 group ,P<0.05 all; 2. Myocardial cells lined up in order and there was no hypertrophy in group DM1; but those in group DM2 showed cells loosely lined up and hypertrophy under light microscope; 3 Immunohistochemical assay indicated that expression of SUMO1 in group DM2 and DM1 group were significantly higher than those of healthy control group [(44.5±1.1) vs. (27.2±2.2) vs. (21.7±3.0)], and of group DM2 was significantly higher than that of DM1 group (P<0.01 all); expression of TNF-α in group DM2 and group DM1 were significantly higher than that of healthy control group [(27.5±1.5) vs. (20.2±2.7) vs. (13.1±1.6)], and of DM2 group was significantly higher than that of group DM1 (P<0.01 all)；expression of NF-κB in group DM2 and group DM1 were significantly higher than that of healthy control group [（30.1±1.7）vs.40.7±1.5）vs.（16.0±2.6）], but of group DM1 was significantly higher than that of group DM2 (P<0.01 all). Conclusion: There are obvious metabolic disorders of glucose and lipid in T2DM rats, and complicated morphological changes of myocardial tissues similar to myocardial lesions in DM humans; the expressions of SUMO1, NF-κB and TNF-α significantly increase, suggest SUMO1 takes part in inflammatory reaction mediated by NF-κB, TNF-α in myocardial lesion of rat with T2DM，and may inhibit NF-κB, possesses effect of protect myocardium.

AIM:To establish the assessing method of traditional Chinese medicine chromatogrphic fingerprints by the involution similarity and quantitative involution similarity and its application. METHODS: The fingerprint of Ginkgo biloba L.extract(GBE) was evaluated by the involution similarity and quantitative involution similarity,and the contrast studies were implemented by other appraisal targets,eg.the included angle cosine S_F,the correlation coefficient r and the indexes with quantitative characteristic,such as the apparently quantitative similarity R%、the average mass percent M%,etc.. RESULTS: The involution similarity could reflect the similarity change in each sample from different bathes and be of quantitative function.The quantitative involution similarity and quantitative weighted similarity could perfectly reflect the changes in contents of the consituents of GBE. CONCLUSION: The involution similarity and quantitative involution similarity can qualitatively and quantitatively assess the similarity between sample and the referential fingerprint.They are certainly the novel method of evaluating the traditional Chinese medicine chromatographic fingerprints.

Objective To study the protective effects of Tianma xingnao capsule on brain blood circulation in mice and rats.Methods The arteria carotis externa was ligated,and arteria carotis interna was kept.The left carotid artery was exposed and placed over an ultrasonic flow probe.The brain flow volume was recorded with an Ultrasonic Volume.The pursiness time of mice was observed to evaluate the effect of Tianma xingnao capsule on cerebral ischemia and anoxia.The permeability of normal blood-cerebral barrier was investigated in mice.Results Tianma xingnao capsule significantly increased brain blood flow and prolonged mouse pursiness time.Tianma xingnao capsule at a dose of 4 g/kg significantly elevated the content of Evans in mouse brain.Conclusions Tianma xingnao capsule can increase the brain blood flow in rats and improve brain blood circulation in mice.It is obviously advantageous to protect against cerebral ischemia and hypoxia.

BACKGROUND: To apply mouse anti-human cTnI monoclonal antibody as the drug vector in the treatment and diagnosis of myocardial injury, it is important to degrade the immunity of murine antibody and overcome human anti-mouse reaction. Humanization has been applied as an attempt to resolve this problem.OBJECTIVE: To clone murine anti-cTnI Fab fragment and analyse the nucleotide and deduced amino acid sequences.DESIGN: Single sample study.SETTING: An institute of cardiovascular disease under a medical university-affiliated hospitalMATERIALS: The study was conducted in the Institute of Cardiovascular Diseases, First Affiliated Hospital of Nanjing Medical University from January 2003 to May 2004. The hybridoma cell line JS200202 which secrets the anti-cTnI monoclonal antibody was provided by Institute of Cardiovascular Disease, First Affiliated Hospital of Nanjing Medical University.METHODS: IgG heavy chain primers and κ light chain primers of amplified mouse were designed. Total RNA was extracted from hybridoma cells which secrete cTnI. Reverse transcription polymerase chain reaction(RT-PCR) was amplified. Cloning and subsequent sequence analysis of the Fab fragment was performed. The deduced amino acid sequence was compared and analysed with previously published sequences.MAIN OUTCOME MEASURES: Heavy chain Fd segment and κ light chain gene sequence and its subgroups.RESULTS: A band of approximate 700 and 800 base pairs were amplified using IgG heavy chain primers and κ light chain primers respectively. Sequence analysis indicated that the deduced amino acid sequences were in consistent with the characterization of the amino acid in the murine IgGl Fab fragment(GenBank accession NO AY484430, AY484431; Protein Bank accession NO AAR83243, AAR83244).CONCLUSION: A complete murine anti-cTnI Fab fragment was obtained in this study, which may provide basis for the production of the chimeric anti-cTnI antibody.

Objective: To study expressions of small ubiquitin-related modifier protein（SUMO）4 (SUMO4), nuclear factor (NF)- κB and inhibitory factor of NF-κB (IκB) in kidneys of rats with type 2 diabetes mellitus (T2DM). Methods: A total of ten 40-week-old male Goto-Kakizaki (GK) rats （with spontaneous diabetes mellitus）of specific-pathogen free (SPF) grade, and ten 40-week-old male Wistar rats of SPF grade were selected. The lesion of renal tissue was observed by hematoxylin eosin (HE) staining. Expressions of SUMO4, NF-κB and IκB in renal tissue were observed by immunohistochemistry methods. Results: In the GK rats, glomerular capillary ball hypertrophy, basilar membrane slightly thickening; glomerular mesangial cells hyperplasia, hypertrophy and renal tubular epithelial cells hypertrophy were observed. Compared with normal Wistar rats, expression levels of NF-κB [(0.232±0.034) vs. (0.634±0.058)], IκB [(0.242±0.027) vs. (0.712±0.078)] and SUMO4 [(0.160±0.031) vs. (0.545±0.045)] significantly increased in renal tissue of GK rats (P<0.01 all). Conclusion: Compared with Wistar rats, expressions of NF-κB, IκB and SUMO4 significantly increase in renal tissue of GK rats, suggesting that SUMO inhibiting transcriptional activity of NF-κB may exist in kidneys of T2DM rats. Therefore, sumoylation may be a new therapeutic target for inhibit renal microvascular lesion of diabetic disease.

Objective To investigate effects of preoperative application of parecoxib on postoperative analgesia and coagulation function in neurosurgical patients.Methods A total of 90 patients (38 males and 52 females,ASA physical status Ⅰ or Ⅱ) undergoing crainotomy were randomly divided into two groups(n=45): parecoxib group (group P) and control group (group C).At 30 min before operation,group P received intravenous injection of parecoxib 40 mg (5 ml),group C intravenous injection of saline 5 ml.Postoperative patient-controlled intravenous analgesia (PCIA) was performed in all patients.PCIA formula of sufentanil 2 μg/kg+tropisetron 0.2 mg/kg,were diluted with normal saline to 120 ml.The visual analogue scale (VAS),the total and effective PCIA pump compressions,Ramsay sedation scale of 2,4,16,24,48 h after operation were recorded.Coagulation function was measured before and 2 h,48 h after parecoxib administration.Meanwhile,adverse reactions were recorded.Results Comparion of VAS between the two groups was made within 48 h after surgery,the total and effective PCIA pump compressions,were much more in group C than in group P (P<0.05).Ramsay sedation scale of group C was higher than that in group P at 2 h after operation.There were no significant differences in coagulation function.And the percentage of patients′ adverse effects in group P was lower than that in group C (P<0.05).Conclusion Parecoxib,as an analgesic,can enhance analgesic effect of sufentanil PCIA.Not only does it reduce the amount of sufentanil and incidence of adverse reactions,but also it has no significant effect on blood coagulation function.

Objective To study the anti-inflammatory,antitussive,and analgesia effects of Jinhua qingre capsules.Methods The anti-inflammatory effect was assessed by the methods include xylene-induced mouse auricular swelling and histamine-induced pigment oozing from skin vessel in rats;The antitussive and analgesic effect were assessed by ammonia water induced cough model and acetic acid-induced twisting method.Results In anti-inflammation experiment,the high dose (12 g·kg-1) and moderate (6 g·kg-1) groups of Jinhua qingre capsules showed significant inhibitory effect on auricular swelling and significant difference compared with control group (P ＜ 0.01,P ＜ 0.05.);the high dose (10 g· kg-1) and moderate dose (5 g·kg-1) groups of Jinhua Qingre capsules play a marked inhibitory role in the increase in mouse peritoneal capillary permeability (P ＜ 0.01,P ＜ 0.05).In antitussive experiment,high dose (12 g· kg-1) and moderate dose (6 g· kg-1) groups had significant inhibitory effect on cough caused by ammonia water (P ＜ 0.01,P ＜ 0.05) compared with control group.In analgesic experiment,the high dose (12 g·kg-1),moderate dose (6 g·kg-1),and low dose (3 g·kg-1) groups effectively reduced the writhing frequency of mice (P ＜ 0.01,P ＜ 0.05).Conclusion Jinhua qingre capsules have potential effects on anti-inflammatory,antitussive,and analgesic.