Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.

Objective To investigate the clinical features and characteristic of auxiliary examination of neuralgic amyotrophy(NA).Methods8 patients with idiopathic neuralgic amyotrophy were analyzed retrospectively with literature review.Results and ConclusionAntecedent events were found before attack in 5 patients.Serious pain,which was followed by paresis and atrophy,as initial symptom,was found in 7 patients.Damage in the upper and middle trunk distribution occurred most frequently.Elevated liver enzymes were found during the early phase of their attacks in 2 patients.CSF analysis was performed in 5 patients,and mildly elevated protein was found in two.5 patients had been misdiagnosed as cervical spondylosis.After medical and/or physical therapy(6 patients with corticosteroid and physical therapy,2 only with physical therapy),all the patients had been followed up for 2～13 months.Muscle power was recovered in 67% of all the patients,partially recovered in 33%.

Objective To evaluate the surgical techniques and clinical efficacy of retroperitoneoscopic ligation of renal lymphatic vessels outside adipose capsule and inside adipose capsule for comparison in the treatment of chyluria.Methods Retrospective anal-ysis of the retroperitoneal laparoscopic adipose capsule of kidney pedicle lymphatic duct ligation(group Ⅰ)and adipose capsule pathways in the kidney totally or subtotally free renal pedicle lymphatic ligation(group Ⅱ)and 1 1 1 patients with chyluria clinical data,compared two groups of operation time,bleeding volume,the rate of analgesia,postoperative gastrointestinal function recovery time,drainage time,postoperative recovery time,postoperative hospital stay and postoperative complications and other index differ-ence.Results Retroperitoneal laparoscopic operation group of adipose capsule in operation time,bleeding volume,postoperative an-algesia,postoperative gastrointestinal function recovery time,drainage time,postoperative recovery time,postoperative hospital stay and postoperative complications were better than the adipose capsule of kidney totally or subtotally free operation group,the differ-ence was statistically significant(P<0.05).Conclusion Retroperitoneal laparoscopic renal adipose capsule of kidney pedicle lym-phatic disconnection for the treatment of chyluria effect,and adipose capsule operation ways,the method less trauma,quicker recov-ery,can completely replace the renal adipose capsule total or Sub-total of free kidney pedicle lymphatic disconnection operation,wor-thy of clinical application.

Objective To investigate the safety and effect of high intensity focused ultrasound ablation(HIFU) on residual liver cancer in patients after radiotherapy. Methods Twenty liver cancer patients with twenty five residual tumors after three-dimensional conformal radiotherapy or stereotactic radiotherapy received HIFU ablation. Liver function and periphery blood cell counts were performed before HIFU and at 1 week after HIFU in all patients. Enhanced MRI and α-fetoprotein (AFP) level were performed before HIFU and at 2 weeks after HIFU to evaluate the effect of HIFU ablation. The survival of all patients was assessed by the Kaplan-Meier method. Results The mean follow-up time was (12.6 ± 8.0)months. The median survival time and 1-year survival rate were 22 months and 87.5% respectively. No skin burns were observed in all patients. As compared with before HIFU,there were no significant differences in the levels of albumin,alanine transarninase, aspartate transarninase, total bilirubin, direct bilirubin, blood red cell counts,blood white cell counts and blood platelet counts at 1 week after HIFU (paired t test, P =0. 156,0. 356,0. 203,0.659,0. 531,0. 519,0. 310,0. 346, respectively). Significant difference in AFP level of 9 patients with AFP＞20 μg/L was observed before HIFU and 2 weeks after HIFU (paired t test, P =0.030). Among 25 residual liver tumors,sixteen with complete ablation and six with ablation volume of ≥80% were observed by enhanced MRI at 2 weeks after HIFU. Conclusions HIFU may be a safe and effective new method to ablate residual liver cancer after three-dimensional conformal radiotherapy or stereotactic radiotherapy.

An HPLC method for the determination of geniposide concentration in mouse plasma was developed and the pharmacokinetics after intranasal administration of Xingnaojing microemulsion (XNJ-M) and mPEG2000-PLA modified Xingnaojing microemulsion (XNJ-MM) were investigated. Eighty mice were treated by XNJ-M and XNJ-MM nasally. The plasma samples were collected at different times and the drug in samples was detected by HPLC. The pharmacokinetic parameters were calculated by the software of Kinetica. The pharmacokinetic parameters of geniposide of XNJ-M were C(max) (4.36 +/- 2.69) mg x L(-1), t(max) 1 min, MRT (29.73 +/- 4.54) min, AUC (53.63 +/- 14.03) mg x L(-1) x min. The pharmacokinetic parameters of geniposide of XNJ-MM were C(max) (9.75 +/- 4.14) mg x L(-1), t(max) 1 min, MRT(22.34 +/- 2.90) min, AUC (131.87 +/- 40.13) mg x L(-1) x min. Geniposide can be absorbed into blood in a higher degree after intranasal administration with XNJ-MM compared to XNJ-M, which maybe caused by its less irritating and more absorption.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Emulsions ; Iridoids ; blood ; pharmacokinetics ; Lactic Acid ; chemistry ; Male ; Mice ; Polyesters ; Polyethylene Glycols ; chemistry ; Polymers ; chemistry

Objective To investigate the clinical efficacy and feasibility of 125I seeds implantation combined with high intensity focused ultrasound (HIFU) for portal vein tumor thrombus (PVTF) from HCC.Methods Nineteen cases of PVTT from HCC (17 males,2 females;average age:50 years,stage:Ⅰ-Ⅲ)undergoing 125I seeds implantation combined with HIFU from March 2011 to October 2013 were retrospectively studied.The radioactivity of each 125I seed was 18.5 MBq and the matched peripheral dose was 90-120 Gy.HIFU parameters were set as follows:ultrasound wave frequency,0.85 mHz;output power,300-400 W;total treatment duration,33-70 min.During the follow-up of 3-24 months,the change of PVTT size,postoperative complications and survival rate were recorded.Kaplan-Meier method was used to analyze the survival rate.Results The treatment was successfully completed in all patients and no serious complications were observed.After 1 month of follow-up,the tumor thrombi shrank in all patients.Among 19 patients,9 achieved PR,4 SD,6 PD.The one-year and two-year survival rates were 47.4％ and 7.9％,respectively.The mean survival was 11.6±3.0 months (range:3-24 months).The median postoperative survival for stages Ⅰ,Ⅱ,Ⅲ was 13.5,7.0,4.0 months,respectively.Conclusion 125I seeds implantation combined with HIFU could be a safe and effective therapy for PVTT from HCC.

Objective To investigate the effects of mesenchymal stem cells (MSCs) transplantation combining with vascular endothelial growth factor (VEGF) gene therapy on myocardium rebuilding,angiogene- sis,and heart function improvement in rats with myocardial infarction.Methods SD rat MSCs were isola- ted,cultured in vitro,labeled with BrdU and transfected by Ad.VEGF gene.Four weeks after left anterior descending artery was ligated to created rat myocardial infarction,cardiac function was examined with echocar- diography,rats were randomly divided into four groups (n=10 in each group):GroupⅠ:MSCs/Ad.VEGF implantation;GroupⅡ:MSCs implantation;GroupⅢ:Ad.VEGF injection;GroupⅣ:Control.MSCs dif- ferentiation was observed 4 weeks after transplantation.Immunohistochemistry and angiogenesis were observed. Echocardiography was performed to detect the effects on heart function.Results MSCs labeled with BrdU could be identified in host hearts in groupⅠandⅡ,most of them positively stained with cTnT antibody. Echocardiography indicated that the improvement of the LVEF value in groupⅠwas more significant than that in the other three groups (P＜0.01,respectively).Some cells were incorporated into the coronary capillaries in the infarcted region.The capillary density in groupⅠwas higher than that in the other three groups (P＜0.01,respectively).Conclusion MSCs implantation combining with VEGF gene therapy can obviously re- pair damaged myocardium and enhance the angiogenesis in ischemic heart tissue.

Objective To explore the effect of fingering expansion anus on the constipation of high risk of cardiovascular disease patients.Methods Total 84 patients in the cardiovascular unit were randomly divided into the treatment group(n=42) and control group (n=42).Patients in the treatment group were treated with fingering expansion anus and used Glycerine Enema (20ml),while the control group were only given Glycerine Enema(20 ml).Curative effect was compared after treatment.Results The effective rate of treatment group was 88.10％ and that of control group was 64.29％,and the difference was statistically significant (H=8.456,P＜0.01).Conclusions Fingering expansion anus is an effective and economic method for the high risk of cardiovascular disease patients with constipation.

of glucose and insulin are also marked after congee intake.

Objective To establish an extensively drug-resistant Acinetobacter baumannii (XDR-AB) infection model using Caenorhabditis elegans (C.elegans),and evaluate the effect of efflux pump inhibitors(EPIs) on reversal of ciprofloxacin resistance in XDR-AB.Methods XDR-AB infection model of C.elegans was established,six EPIs(CCCP,PAβN,NMP,omeprazole,reserpine,and verapamil)combined with ciprofloxacin were used to treat the infected model,the survival rate of C.elegans was recorded to evaluate the in vivo activities of drugs,toxicity test and in vitro drug susceptibility test were also performed.Results Lethal effect of different concentrations of XDR-AB on C.elegans was varied,5 × 106 CFU/mL of XDR-AB was selected to infect C.elegans.C.elegans survival test showed that survival curves of C.elegans infected with XDR-AB for 3 hours and curves of control group (polymixin B was added) were not significantly different (x2 =3.154,P＞0.05);compared with control group,survival curves of C.elegans infected with XDR-AB for 6 hours or 9 hours were significantly different (both P＜0.001),but 6 hours and 9 hours were not significantly different(x2 =0.669,P＞0.05),6 hours was chosen as the duration of infection,36 hours was appropriate for the duration of antimicrobial therapy.Ciprofloxacin with EPIs for infection model revealed that low concentration of PAβN,NMP,omeprazole,and reserpine could improve the survival rate of C.elegans by 30％-40％,15％-20％,20％-30％,and 20％ respectively,high concentration of verapamil could improve the survival rate of infected C.elegans by about 30％.In vitro susceptibility test and toxicity test results showed that ciprofloxacin combined respectively with CCCP,omeprazole,and verapamil could reduce minimum inhibitory concentration(MIC) to the original 1/4,combined respectively with PAβN,NMP,and reserpine could reduce MIC to the original 1/2,CCCP had the best bacterial inhibitory effect in vitro,but the toxicity was large,and was not suitable for the study of pharmacodynamics in vivo.Conelusion The infection model of C.elegans XDR-AB is initially and successfully established,which is used to evaluate the efficiency of six EPIs for reversing ciprofloxacin resistance.