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1.
Chinese Journal of Surgery ; (12): 164-168, 2022.
Article in Chinese | WPRIM | ID: wpr-935596

ABSTRACT

Objective: To examine the effectiveness and safety of application of the ureteral access sheath in the treatment of middle or lower ureteral calculi in patients with large-volume benign prostatic hyperplasia above grade Ⅲ, which is expected to avoid the simultaneous or staged treatment of benign prostatic hyperplasia via eliminate the difficult angle and resistance of ureteroscopy caused by severe prostatic hyperplasia. Methods: From April 2018 to December 2020, the clinical data of 27 patients with massive benign prostatic hyperplasia above grade Ⅲ and middle and lower ureteral calculi treated with indwelling ureteral access sheath plus ureteroscopy holmium laser lithotripsy at Department of Urology, Zhejiang Quhua Hospital were retrospectively analyzed and followed up. All the patients were male, aged (69.7±12.8) years (range: 55 to 87 years). Prostate volume measured by transrectal ultrasound was (94.8±16.2) cm3 (range: 85 to 186 cm3). The ureteral access sheath was indwelled in advance, and then the semirigid ureteroscopy was introduced through the working channel of the sheath. Holmium laser lithotripsy was performed, and intraoperative and postoperative complications were recorded. Urinary abdominal plain X-ray or CT urography were performed at 1-and 2-month postopaerative to evaluate the residual stones and clinical efficacy. Results: The ureteral access sheath was placed and holmium laser lithotripsy under a semirigid ureteroscopy was performed successfully in all the 27 patients. In 2 patients, a second session of auxiliary procedure was required due to the large load of preoperative stones and residual stones after surgery, among whom 1 patient received extracorporeal shock wave lithotripsy and 1 patient underwent extracorporeal shock wave lithotripsy plus ureteroscopic lithotripsy. The stone free rate at 1-and 2-month postoperative were 92.6% (25/27) and 100% (27/27), respectively. There were no severe complications such as ureteral avulsion and perforation, perirenal hematoma, septic shock, severe hematuria, urinary retention, iatrogenic ureteral stricture occurred during and after the surgery. The ureteral calculus was wrapped by polyps heavily in 1 patient, he was diagnosed as ureteral stenosis 1 month postoperative, receiving laparoscopic resection of ureteral stricture plus anastomosis 3 months postoperative. Conclusions: In the operations of middle and lower ureteral calculi in patients with large-volume prostatic hyperplasia above grade Ⅲ, the ureteral access sheath can be placed first to effectively eliminate the difficult angle and resistance of ureteroscopy caused by severe prostatic hyperplasia, and then semirigid ureteroscopic lithotripsy can be safely performed. It could avoid the treatment of benign prostatic hyperplasia at the same time or by stages.


Subject(s)
Aged , Aged, 80 and over , Humans , Male , Middle Aged , Lithotripsy , Lithotripsy, Laser , Prostatic Hyperplasia/complications , Retrospective Studies , Treatment Outcome , Ureteral Calculi/surgery , Ureteroscopy
2.
Chinese Traditional and Herbal Drugs ; (24): 1211-1219, 2018.
Article in Chinese | WPRIM | ID: wpr-852161

ABSTRACT

Melastoma dodecandrum is commonly used in the folk. It has been widely used in China by She, Yao, Miao, and other ethnic minorities, which has the effects of blood activating and hemostasis, elimination of swelling and stasis, and heat-clearing and detoxifying. In this review, the relevant researches about M. dodecandrum were summarized from the textual research, resource distribution, standards, pharmacological activity, chemical composition and clinical application aspects, which provides a theoretical basis for the further study and clinical application of M. dodecandrum.

3.
Chinese Pharmaceutical Journal ; (24): 119-123, 2017.
Article in Chinese | WPRIM | ID: wpr-858839

ABSTRACT

OBJECTIVE: To investigate the correlation of blood concentration and drug dose and reducing lipid efficacy of tectorigenin in atherosclerosis rabbits. METHODS: Forty-eight Japanese white rabbits were randomly divided into six groups, the normal control group, model group, high-, middle-and low-dose (50, 25, 12.5 mg·kg-1·d-1) tectorigenin groups and atorvastatin (5 mg·kg-1· d-1) group. The normal control group was fed with routine forage, and the other groups were fed with high fat forage for the establishment of atherosclerosis model. After 8 weeks, tectorigenin blood concentration was detected by HPLC. After 12 weeks, blood lipids were detected with automatic biochemical analyzer. We sheared the thoracic and abdominal aorta to observe the formation of atherosclerotic plaque and make pathologic examination. RESULTS: Steady-state blood concentration increased with the increase of drug dose, there was a positive relationship (r = 0.969 1) between them, but not the same proportion. After treatment, serum total cholesterol (TC), triglyceride(TG), low density lipoprotein cholesterol (LDL-C) levels were decreased and high density lipoprotein cholesterol (HDL-C) were increased in the tectorigenin each dose group, but the effect was weaker than atorvastatin group. CONCLUSION: Tectorigenin has obvious effect on reducing blood lipid and anti-atherosclerosis in rabbits, and it is related to the blood concentration and the drug dose.

4.
Chinese Traditional and Herbal Drugs ; (24): 266-271, 2017.
Article in Chinese | WPRIM | ID: wpr-853028

ABSTRACT

Objective: To evaluate therelease characteristicsin vitro, pharmacokinetics in rabbits and in vivo-in vitro correlation of tectorigenin floating sustained-release tablets (TFSRT). Methods: The release characteristics of TFSRTin vitro was detected with HPLC in the artificial gastric fluid. Six Japanese Giant Ear Rabbits as self crossover control, which were given TFSRT and suspension liquid (200mg). The concentration of tectorigenin in plasma was determined with HPLC and the data were processed with PKsolver 2.0 software. Results:The cumulative releaserate of TFSRTin vitro was over 70% in 10 h.The pharmacokineticsin rabbits showed that TFSRT and tectorigenin suspension liquid conformed to the single compartment model and the pharmacokinetic parameters were obtained: tmax: (2.809±0.371) and (0.442±0.138)h, Cmax: (6.317±1.337) and (9.662±2.759) μg/mL, AUC0-t: (74.156±10.420) and (57.059±13.309) μg∙h/mL. The relative bioavailability of TFSRT was (134.63±27.94)%, so there was significant difference between them. Conclusion: TFSRT can release slowly, so it increase the relative bioavailability significantly. The correlation between the absorption in vivo and release in vitro is fine (r=0.9879), so the release rate in vitro can control the quality of TFSRT.

5.
China Journal of Chinese Materia Medica ; (24): 298-302, 2017.
Article in Chinese | WPRIM | ID: wpr-230956

ABSTRACT

To investigate the preparation technology and release mechanism of tectorigenin intragastric floating sustained-release tablets. The tablet was produced by wet granulation compression method, with hydroxypropyl methyl cellulose (HPMCK15M), cross-linked polyvinyl pyrrolidone (PVPP), octadecanol and sodium bicarbonate as excipient. The prescriptions were screened and optimized by orthogonal experimental design with in vitro floating capacity and drug release characteristics as the evaluation indexes. The optimization results were as follows: tectorigenin 33.3%, HPMCK15M 16.7%, PVPP 20.0%, octadecanol 13.3%, sodium bicarbonate 5%, and starch gel 10.7%. The prepared tablet can be floated within 10 s in the artificial gastric juice, lasting for 12 h in vitro, with a cumulative release rate of 70% in 10 h. The analysis of Rritger-Peppas equation showed that the sustained-release tablet had two advantages of both drug diffusion and skeleton dissolution. The tablet had good appearance and compressibility, as well as favorable floating capacity and drug release characteristics.

6.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 1103-1107, 2014.
Article in Chinese | WPRIM | ID: wpr-294339

ABSTRACT

<p><b>OBJECTIVE</b>To explore the in vitro anti-tumor effect and mechanism of dendritic cell (DC) tumor vaccine induced by astragalus polysacharin (APS).</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMCs) isolated from human peripheral blood. DCs obtained from human peripheral blood were cultivated and added with culture solution for in vitro inducing them to immature DCs. On the 5th day of culture, 100 microg/mL (as the final concentration) APS was added to cells in the APS group. DCs were induced to mature in the cytokine groups by adding 20 ng/mL rhTNF-alpha (as the final concentration). Changes of morphology and phenotype of DCs were observed. Mature DCs were sensitized with tumor antigen SGC-7901 and co-cultured with allogeneic T cells. The proliferative function of T lymphocytes was detected by MTT assay. Levels of IL-12 and IFN-gamma in co-cultured supernatant were detected by ELISA. Cytotoxic lymphocytes (CTL) activated by DC were co-cultured with tumor cell SGC-7901. The specific killing capacity of CTL to target cells was detected by LDH release assay.</p><p><b>RESULTS</b>The morphological observation and phenotypic identification of APS induced DCs were in accordance with the characteristics of mature DCs. APS induced mature DCs could stimulate the proliferation of allogeneic T lymphocytes. The proliferation index of T cells increased with increased ratio of stimulator cells to effector cells (P < 0.05). Levels of IL-12 and IFN-gamma in co-culture supernatant significantly increased in a time-dependent manner (P < 0.05). CTL cells activated by sensitization of DCs could significantly kill tumor cells, and the killing effect increased along with increased effector-to-target ratio.</p><p><b>CONCLUSION</b>APS could in vitro induce DCs to mature, promote its antigen-presenting capacity, effectively activate CTLs, and enhance anti-tumor function of the organism.</p>


Subject(s)
Humans , Antigen-Presenting Cells , Cell Biology , Allergy and Immunology , Cancer Vaccines , Allergy and Immunology , Cell Line , Cell Proliferation , Coculture Techniques , Dendritic Cells , Cell Biology , Allergy and Immunology , Drugs, Chinese Herbal , Pharmacology , Interferon-gamma , Allergy and Immunology , Interleukin-12 , Allergy and Immunology , Leukocytes, Mononuclear , Cell Biology , Allergy and Immunology , Lymphocyte Activation , T-Lymphocytes, Cytotoxic , Cell Biology
7.
Acta Physiologica Sinica ; (6): 667-674, 2014.
Article in Chinese | WPRIM | ID: wpr-255988

ABSTRACT

The present study is aimed to investigated the firing activity of pyramidal neurons and interneurons in the medial prefrontal cortex (mPFC) in rats with bilateral intraventricular injection of 5,7-dihydroxytryptamine (5,7-DHT) by using in vivo extracellular recording. The results showed that the injection of 5,7-DHT reduced the 5-hydroxytryptamine (5-HT) levels in the mPFC and dorsal raphe nucleus in the rats. The firing rate of mPFC pyramidal neurons in rats with 5,7-DHT injection was significantly higher than that of normal rats, and the firing pattern of these neurons also changed significantly towards a more burst-firing, while the injection decreased the firing rate of mPFC interneurons and changed the firing pattern of the interneurons towards a more irregular. These results indicate that the lesions of the serotonergic neurons lead to the changes in the firing activity of mPFC pyramidal neurons and interneurons, suggesting that serotonergic system plays an important role in the regulation of the neuronal activity in the mPFC.


Subject(s)
Animals , Rats , 5,7-Dihydroxytryptamine , Pharmacology , Action Potentials , Dorsal Raphe Nucleus , Cell Biology , Injections, Intraventricular , Interneurons , Prefrontal Cortex , Cell Biology , Pyramidal Cells , Serotonin , Metabolism
8.
Biomedical and Environmental Sciences ; (12): 569-576, 2012.
Article in English | WPRIM | ID: wpr-320397

ABSTRACT

<p><b>OBJECTIVE</b>To develop a new technique for assessing the risk of birth defects, which are a major cause of infant mortality and disability in many parts of the world.</p><p><b>METHODS</b>The region of interest in this study was Heshun County, the county in China with the highest rate of neural tube defects (NTDs). A hybrid particle swarm optimization/ant colony optimization (PSO/ACO) algorithm was used to quantify the probability of NTDs occurring at villages with no births. The hybrid PSO/ACO algorithm is a form of artificial intelligence adapted for hierarchical classification. It is a powerful technique for modeling complex problems involving impacts of causes.</p><p><b>RESULTS</b>The algorithm was easy to apply, with the accuracy of the results being 69.5%±7.02% at the 95% confidence level.</p><p><b>CONCLUSION</b>The proposed method is simple to apply, has acceptable fault tolerance, and greatly enhances the accuracy of calculations.</p>


Subject(s)
Humans , Infant, Newborn , Algorithms , Artificial Intelligence , China , Epidemiology , Environmental Exposure , Models, Biological , Neural Tube Defects , Epidemiology , Risk Factors
9.
Chinese Journal of Epidemiology ; (12): 431-435, 2011.
Article in Chinese | WPRIM | ID: wpr-273172

ABSTRACT

Objective To analyze the results of application on China Infectious Diseases Automated-alert and Response System(CIDARS)and for further improving the system. Methods Amount of signal, proportion of signal responded, time to signal response, manner of signal verification and the outcome of each signal in CIDARS were descriptively analyzed from July 1,2008to June 30, 2010. Results A total of 533 829 signals were generated nationwide on 28 kinds of infectious diseases in the system. 97.13% of the signals had been responded and the median time to response was 1.1 hours. Among them, 2472 signals were generated by the fixed-value detection method which involved 9 kinds of diseases after the preliminary verification, field investigation and laboratory tests. 2202 signals were excluded, and finally 246 cholera cases, 15 plague cases and 9H5N1 cases as well as 39 outbreaks of cholera were confirmed. 531 357 signals were generated by the other method - the 'moving percentile method' which involved 19 kinds of diseases. The average amount of signal per county per week was 1.65, with 6603 signals(1.24%)preliminarily verified as suspected outbreaks and 1594 outbreaks were finally confirmed by further field investigation. For diseases in CIDARS, the proportion of signals related to suspected outbreaks to all triggered signals showed a positive correlation with the proportion of cases related to outbreaks of all the reported cases (r=0.963, P<0.01). Conclusion The signals of CIDARS were responded timely, and the signal could act as a clue for potential outbreaks, which helped enhancing the ability on outbreaks detection for local public health departments.

10.
Chinese Journal of Epidemiology ; (12): 436-441, 2011.
Article in Chinese | WPRIM | ID: wpr-273171

ABSTRACT

Objective To analyze the pilot results of both temporal and temporal-spatial models in outbreaks detection in China Infectious Diseases Automated-alert and Response System (CIDARS)to further improve the system. Methods The amount of signal, sensitivity, false alarm rate and time to detection regarding these two models of CIDARS, were analyzed from December 6,2009 to December 5,2010 in 221 pilot counties of 20 provinces. Results The sensitivity of these two models was equal(both 98.15%). However, when comparing to the temporal model, the temporal-spatial model had a 59.86% reduction on the signals(15 702)while the false alarm rate of the temporal-spatial model(0.73%)was lower than the temporal model(1.79%), and the time to detection of the temporal-spatial model(0 day)was also 1 day shorter than the temporal model.Conclusion Comparing to the temporal model, the temporal-spatial model of CIDARS seemed to be better performed on outbreak detection.

11.
National Journal of Andrology ; (12): 707-711, 2011.
Article in Chinese | WPRIM | ID: wpr-305802

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the association of the IL-6 -572C > G polymorphism with the risk of prostate cancer (PCa) in the Chinese Han population in Jiangsu and Anhui area.</p><p><b>METHODS</b>We obtained peripheral blood genome DNA from 200 PCa patients and 279 age-matched PCa-free healthy controls, analyzed the site polymorphism of IL-6 -572C > G with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, and studied the correlation of different genotypes with the susceptibility to PCa.</p><p><b>RESULTS</b>The subjects that carried the CCGG genotype had a risk of PCa 2.46 times that of the CC genotype carriers (95% CI = 1.41-4.29), and 2.47 times that of the CC/GC genotype carriers (95% CI = 1.47-4.17). This risk was significantly increased among the following subgroups of CCGG genotype carriers: age > 70 yr (OR = 3.06, 95% CI: 1.44-6.49), BMI > 23 kg/m2 (OR = 3.72, 95% CI: 1.79-7.74), no cigarette smoking (OR = 2.96, 95% CI: 1.30-6.72), alcohol drinking (OR = 2.73, 95% CI: 1.28-5.79), with a family history of cancer (OR = 6.67, 95% CI: 1.50-29.69).</p><p><b>CONCLUSION</b>In the Chinese Han population in Jiangsu and Anhui area, IL-6 -572C > G polymorphism is associated with the susceptibility to PCa, and GG might be a susceptible genotype to PCa.</p>


Subject(s)
Aged , Humans , Male , Asian People , Genetics , Case-Control Studies , China , Epidemiology , Gene Frequency , Genetic Predisposition to Disease , Genotype , Interleukin-6 , Genetics , Polymorphism, Single Nucleotide , Prostatic Neoplasms , Epidemiology , Genetics
12.
Biomedical and Environmental Sciences ; (12): 167-172, 2010.
Article in English | WPRIM | ID: wpr-360607

ABSTRACT

<p><b>OBJECTIVE</b>To predict neural tube birth defect (NTD) using support vector machine (SVM).</p><p><b>METHOD</b>The dataset in the pilot area was divided into non overlaid training set and testing set. SVM was trained using the training set and the trained SVM was then used to predict the classification of NTD.</p><p><b>RESULT</b>NTD rate was predicted at village level in the pilot area. The accuracy of the prediction was 71.50% for the training dataset and 68.57% for the test dataset respectively.</p><p><b>CONCLUSION</b>Results from this study have shown that SVM is applicable to the prediction of NTD.</p>


Subject(s)
Humans , China , Epidemiology , Neural Tube Defects , Epidemiology , Pilot Projects
13.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 1068-1072, 2009.
Article in Chinese | WPRIM | ID: wpr-242357

ABSTRACT

<p><b>OBJECTIVE</b>To clarify the correlation between primary gout and insulin-resistance (IR), and to observe the effect of serial gout granules (SGGs) on IR in patients with primary gout.</p><p><b>METHODS</b>Pearson's correlation analysis and multiple stepwise correlation analysis were conducted between 60 patients and 60 healthy volunteers in terms of blood pressure (BP), blood levels of uric acid (sUA), fasting plasma glucose (FPG), fasting insulin (FINS), fasting leptin (FL), triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), and C-peptide (CP), body mass index (BMI), waist-to-hip ratio (WHR), insulin resistance index (IRI) and insulin sensitivity index (ISI). Besides, the 60 primary gout patients were equally randomized into two groups, the treatment group treated with SGGs (huzhang gout granule and yinlian gout granule), and the control group treated with diclofenac sodium dual release enteric-coated capsules and Benzbromarone. bove-mentioned indexes in them were assessed before and after 12 weeks of treatment.</p><p><b>RESULTS</b>The levels of sUA, FPG, TC, TG, LDL-C, FINS, CP, FL, IRI, BMI, and WHR were higher while the levels of HDL-C and ISI was lower in gout patients than those in healthy controls (all P < 0.05); Pearson's correlation analysis showed that level of sUA was positively correlated with levels of FPG, FINS, IRI, BMI, TC, TG, LDL-C, CP, FL, systolic pressure and diastolic pressure (r = 0.444, 0.496, 0.660, 0.542, 0.414, 0.467, 0.344, 0.470, 0.419, 0.275, 0.330 respectively, P < 0.05), but negatively correla- ted with levels of ISI and HDL-C (r = -0.569, -0.264 respectively, P < 0.05). Multiple stepwise correlation analysis showed that sUA was also positively correlated with IRI, TG and BMI (r = 5.758, 2.849, 3. 425 respectively, P < 0.05). After 12 weeks of treatment, the lowed range of TC, TG, FINS, FPG and IRI, and the increased amplitude of HDL-C and ISI in the treatment group were superior to those in the control group respectively (P < 0.05).</p><p><b>CONCLUSION</b>Patients with primary gout are low in insulin sensitivity and indicating the existence of IR. SGGs could enhance patients' insulin sensitivity and improve IR to a certain extent.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Case-Control Studies , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Gout , Drug Therapy , Metabolism , Insulin Resistance , Phytotherapy
14.
Journal of Southern Medical University ; (12): 1635-1637, 2009.
Article in Chinese | WPRIM | ID: wpr-282628

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect of beta(3)-adrenoceptor (AR) in regulating resting intracellular Ca(2+) concentration of the ventricular myocytes and investigate the signaling pathway in rats with experimental heart failure.</p><p><b>METHODS</b>Rat models of experimental heart failure were established by ligation of the anterior descending artery, and the myocytes were isolated by enzymatic digestion. The resting intracellular Ca(2+) concentration was determined using laser scanning confocal microscopy (LSCM) in the cells stimulated with 1 micromol/L BRL37344 (a selective beta(3)-AR agonist) alone or in combination with PTX, L-NAME, or methylene blue.</p><p><b>RESULTS</b>In the ventricular myocytes from normal control rats, BRL373444 reduced the resting intracellular Ca(2+) concentration of by 45.5%, while the reduction increased to 59.4% in the cells from rats with heart failure. In combination with L-NAME (10 micromol/L), methylene blue (10 micromol/L), and PTX (2 microg/ml), BRL373444 caused a reduction in resting intracellular Ca(2+) concentration of the ventricle myocytes from normal control rats by 10.1%, 16.9%, and 15.4%, respectively in control group, while the rate was 16.9%, 19.3%, and 11.7% in the heart failure group.</p><p><b>CONCLUSIONS</b>Beta(3)-AR agonist can decrease the resting intracellular Ca(2+) concentration of the ventricular myocytes, but the reduction is smaller in cells from rats with heart failure than in cells of normal rats. This effect is mediated through the PTX-NOS-NO pathway.</p>


Subject(s)
Animals , Male , Rats , Adrenergic Agonists , Pharmacology , Adrenergic beta-3 Receptor Agonists , Calcium , Metabolism , Heart Failure , Metabolism , Pathology , Heart Ventricles , Pathology , In Vitro Techniques , Intracellular Space , Metabolism , Myocytes, Cardiac , Metabolism , Pathology , Rats, Wistar , Receptors, Adrenergic, beta-3 , Metabolism , Rest , Signal Transduction
15.
Journal of Experimental Hematology ; (6): 670-673, 2009.
Article in Chinese | WPRIM | ID: wpr-334048

ABSTRACT

This study was aimed to explore the influence of staphylokinase derivative (SAKD) on the hemoagglutinative and fibrinolytic systems, and to determine the safety of the staphylokinase derivative in application. The normal and model rats each 30 were divided into normal saline, SAKD and rSAK groups. The hemorrhage, bleeding time (BT), blood platelet count (BPC), activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen (Fg), D-dimer (D-D), plasminogen (PLG) and plasmin inhibitor activity (PI) were detected before and after the administration with staphylokinase derivative 0.5 mg/kg body weight, once three days for consecutive 15 days. The results indicated that one case of normal rats with SAKD and two cases of high fat diet model group had mild hemorrhage, all of which showed automatic hemostasis; and 3 cases in rSAK group had mild hemorrhage. And the platelet counting, D-D, PLG and PI in all groups did not significantly change. The rats of high fat diet group treated with SAKD showed the significant extension of APTT, PT and TT times, and the decrease of Fg time (p < 0.05). All the experimental results demonstrated that the influence of SAKD on the hemagglutination of the normal animals was lower, however, which can improve the high-hemagglutination status of the rats with high fat diet. It is concluded that the SAKD at the dosage of this study has the higher safety, which can alleviate the high hemagglutination symptoms of the rats with high fat diet.


Subject(s)
Animals , Male , Rats , Dietary Fats , Fibrin Fibrinogen Degradation Products , Fibrinolysis , Fibrinolytic Agents , Pharmacology , Hemagglutination , Hemostasis , Metalloendopeptidases , Pharmacology , Partial Thromboplastin Time , Platelet Count , Prothrombin Time , Rats, Wistar , Recombinant Fusion Proteins , Pharmacology , Thrombin Time , Thrombolytic Therapy
16.
Journal of Experimental Hematology ; (6): 667-670, 2008.
Article in Chinese | WPRIM | ID: wpr-267914

ABSTRACT

This study was purposed to investigate the angiogenin (ANG) expression in COS-7 cells and its biological activity. The gene of angiogenin was obtained from mononuclear cells of peripheral blood by using RT-PCR and inserted into eukaryotic expression vector of pcDNA3.1. After being transfected into COS-7 cells, the recombinant ANG was identified by Western blot assay. The function of promoting proliferation of ANG to ECV304 cells was detected by MTT method, and its activity of vascularization was analyzed by chick embryo chorioallantois treated by the culture supernatant after transfection with pcDNA3.1-ang. The result showed that recombinant ANG was expressed in COS-7 cells after transfection for 24 to 36 hours. It could specifically react with monoclonal antibody against ANG. The recombinant ANG could obviously promote the proliferation of ECV304 cells. In contrast with the control group, the culture supernatant of pcDNA3.1-ang transfected group could stimulate the angiogenesis in embryo chorioallantois. It is concluded that the ang transiently expresses in COS-7 cells, and its expression product obviously stimulates the cell proliferation and angiogenesis.


Subject(s)
Animals , Humans , Angiogenesis Inducing Agents , Pharmacology , COS Cells , Metabolism , Cell Line , Cell Proliferation , Chlorocebus aethiops , Endothelial Cells , Cell Biology , Genetic Vectors , Genetics , Recombinant Fusion Proteins , Genetics , Pharmacology , Ribonuclease, Pancreatic , Genetics , Pharmacology , Transfection
17.
Journal of Experimental Hematology ; (6): 352-356, 2007.
Article in Chinese | WPRIM | ID: wpr-230269

ABSTRACT

This study was aimed to construct the soluble HLA-A*0201-PR1 complex for preparation of HLA-A*0201-PR1 tetramer. The recombinant HLA-A*0201-BSP (BirA substrate peptide) fusion protein as heavy chain and beta(2)-microglobulin (beta(2) m) as light chain were expressed highly as insoluble aggregates in Escherichia coli and then purified with gel filtration, and the final purity reached above 90%. The two subunits were refolded to form an HLA-A*0201-peptide complex by dilution method in the presence of an antigenic peptide PR1, a HLA-A2-restricted peptide from proteinase 3 (aa 169 - 177, VLQELNVTV). Refolded HLA-A*0201-PR1 complex was biotinylated using a BirA enzyme and purified by anion exchange chromatography on a Q-Sepharose (fast flow) column. The extent of reconstitution of the HLA-A*0201-PR1 complex was analyzed by HPLC gel filtration. The refolded and biotinylated products were detected by Western blot and ELISA with monoclonal antibody BB7.2 that recognized the natural conformations of HLA-A2 and streptavidin. The results showed that the refolded complex was composed of HLA-A*0201-BSP aggregate, HLA-A*0201-PR1 complex and beta(2) m, and reconstitution yields of 18% with PR1 was obtained. Refolded HLA-A*0201-PR1 complex could be confirmed by practical immunological method and biotinylated efficiently. It is concluded that the refolding and biotinylation of HLA-A*0201-PR1 complex is successfully obtained. This work provides the basis for the preparation of HLA-A*0201-PR1 tetramer.


Subject(s)
Humans , DNA Primers , Genetics , Escherichia coli , Genetics , HLA-A Antigens , Genetics , HLA-A2 Antigen , Oligopeptides , Genetics , Metabolism , Protein Binding , Protein Folding , Recombinant Fusion Proteins , beta 2-Microglobulin , Chemistry , Genetics
18.
Chinese Journal of Epidemiology ; (12): 487-490, 2007.
Article in Chinese | WPRIM | ID: wpr-294308

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the diagnostic applicability of human papillomavirus (HPV) liquid chip assay which is based on Luminex XMAP System, and perform a HPV epidemiologic study with the liquid chip in women of Shandong province.</p><p><b>METHODS</b>To detect HPV genotypes on a 96-well plate with the liquid chip which can simultaneously detect and identify 26 common HPV genotypes in a total of 2925 cervical scrapes obtained from gynecological outpatients as well as to analyze the relationship between HPV types and different cervical diseases by studying the distribution of HPV genotypes and pathologic diagnosis.</p><p><b>RESULTS</b>Among 639 cases who performed pathologic/cytological and histological diagnoses, 184 cases are in group of normal cytology, 266 cases in group of, 77 cases in group of cervical intra-epithelial neoplasia (CIN) I, 7 cases in group of CIN I - II, 46 cases in group of CIN I - II, 46 cases in group of CIN I - II and 13 cases in group of cervical cancer. The overall incidence of HPV in our samples is 36.0% (1054/2925) and 23 types of all 26 types on liquid chip are found. The most common genotypes found are HPV-16 (26.75%), HPV-52 (25.75%), HPV-58 (10.47%), HPV-18 (8.87%) and HPV-11 (6.94%). Among all the positive types, 87.32% are high-risk HPV and 13.68% are low-risk HPV genotypes. Both single and multiple types are easily identified, showing 66.22% ( n = 698) single type and 33.78% ( n = 356) multiple types. Of all the 1054 HPV-positive cases, 261 (24.8%) is occupied by women 21 to 25 years of age and progressively lower by older age groups, reaching 4.9% by women between 51 to 67 years old. The incidence of HPV in our samples is 23.37%, 33.08%, 54.54%, 57.14%, 82.61%, 91.30% and 100% for normal cytology, inflammation,CIN I ,CIN I - II, CIN II ,CIN III, and carcinomas specimens, respectively. Infections with more that one virus are common, accounted for 4.89%, 7.14%, 18.18%, 28.57%, 41.30%, 43.37% and 38.46% for normal cytology, inflammation, CIN I, CIN I - II, CIN II, CIN III, and carcinomas specimens, respectively. Based on the criteria of histology and pathology, the sensitivity, specificity, positive-predictive value and negative-predictive value of HPV liquid chip assay for detecting all cases of CIN II, III are 88.57%, 76.63%, 68.89% and 92.16% respectively. Conclusion The common types of HPV infection are 16, 52, 58, 18, 11, 6, 56 and 31. The HPV-positive rate increased along with the increase of grading on cervical lesions. There are more younger women among all the HPV-positive ones. Multiplex HPV genotyping by liquid chip appears to be highly suitable for diagnostic screening as well as the conduction of large-scale epidemiological studies.</p>


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Middle Aged , Young Adult , Uterine Cervical Dysplasia , Epidemiology , Virology , China , Epidemiology , Gammapapillomavirus , Classification , Genetics , Genotype , Human papillomavirus 11 , Classification , Genetics , Human papillomavirus 16 , Classification , Genetics , Human papillomavirus 18 , Classification , Genetics , Human papillomavirus 6 , Classification , Genetics , Oligonucleotide Array Sequence Analysis , Methods , Papillomaviridae , Classification , Genetics , Papillomavirus Infections , Epidemiology , Virology , Uterine Cervical Neoplasms , Epidemiology , Virology
19.
Chinese Journal of Cardiology ; (12): 629-632, 2007.
Article in Chinese | WPRIM | ID: wpr-307232

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the possible association between Tpeak-Tend (Tp-e) interval in surface standard ECG and cardiac events in patients with Brugada syndrome (BrS).</p><p><b>METHOD</b>Tp-e interval in surface standard ECG was compared between BrS patients (n = 23, all males) and paroxysmal supraventricular tachycardia (PSVT) patients (n = 20, all males) as well as between BrS patients with (n = 16) or without (n = 7) cardiac events.</p><p><b>RESULTS</b>There was significant difference in Tp-e interval between BrS patients and PSVT patients [(109.57 +/- 22.86) ms vs. (88.50 +/- 13.08) ms, P < 0.05]. Tp-e interval was also significantly longer in BrS patients with cardiac events (syncope, clinical ventricular fibrillation and induced VF during electrophysiological study) than BrS patients without cardiac events [(118.12 +/- 20.40) ms vs. (90.00 +/- 15.27) ms, P < 0.05] while Tp-e interval was similar between BrS patients without cardiac events and PSVT patients (P > 0.05).</p><p><b>CONCLUSION</b>The prolongation of Tp-e (> or = 120 ms) was associated with higher cardiac events in BrS patients.</p>


Subject(s)
Adult , Humans , Male , Middle Aged , Brugada Syndrome , Diagnosis , Electrocardiography , Risk Factors , Ventricular Fibrillation , Diagnosis
20.
Journal of Experimental Hematology ; (6): 521-524, 2006.
Article in Chinese | WPRIM | ID: wpr-233554

ABSTRACT

Human beta(2)-microglobulin (beta(2)m) is the light chain of major histocompatibility complex (MHC) class I molecule. High-yield production of this protein is a prerequisite to the preparation of MHC class I tetramer. The present study was aimed to obtain recombinant human beta(2)m expressed in Escherichia coli (E. coli) for preparing MHC class I tetramers. For cloning of human beta(2)m gene, a pair of specific primers was designed based on the published sequence of this gene. A 300 bp specific DNA fragment corresponding to the encoding region of beta(2)m lack of the signal peptide sequence was obtained by RT-PCR from the total RNA of human leukocytes. The amplified cDNA was inserted into the IPTG-inducible expression plasmid pET-17b by Nde I and Bam H I sites and its sequence was confirmed by DNA sequence analysis. The recombinant plasmid pET-beta(2)m was transformed to the competent cells of E. coli BL21 (DE3). The results showed that beta(2)m was expressed in the form of inclusion body and amounted to over 32% of total cell proteins after IPTG induction. After washing with triton X-100 and urea, the inclusion body was dissolved with 4 mol/L urea and then purified with Sephacryl S-200 HR, and the final purity reached above 95%. The denatured protein was renatured by dilution method. Western blot assay indicated that the monoclonal antibody against human native beta(2)m could react specifically with the recombinant protein. In conclusion, the human beta(2)m gene was cloned successfully and expressed efficiently in E. coli BL21 (DE3). This work establishes a convenient approach for renaturation and purification of large quantity of recombinant beta(2)m. This provides the basis for the preparation of MHC tetramers.


Subject(s)
Humans , Cloning, Molecular , Escherichia coli , Genetics , Gene Expression , Histocompatibility Antigens Class I , Genetics , Recombinant Proteins , beta 2-Microglobulin , Genetics
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