1.Optimization of Sulfated Technology of Rhizoma Smilacis Glabrae Polysaccharides by Orthogonal Design and Observation of Anti-tumor Activity of Modified Products
Jun QIN ; Guanghai DENG ; Mingchao LUO ; Wei ZHU
Journal of Guangzhou University of Traditional Chinese Medicine 2017;34(2):254-260
Objective To optimize the sulfated modification conditions for Rhizoma Smilacis Glabrae polysaccharides (RSGP),and to investigate the possibility of enhancing the activity of RSGP after sulfating modification.Methods RSGP was modified by cholorosulfonic acid-pyridine.With the yield,carbohydrate content and sulphate substitution degree as the observation indexes,L9 (34)orthogonal design was used to optimize reaction time,reaction temperature and reagent ratio.The degree of sulphate substitution was determined by barium chloride turbitimetry,and the carbohydrate content was detected by sulfuric acid-phenol method.Then the structures of the sulfated modified products were analyzed by infrared radiation (IR) spectrum.Methyl thiazolyl tetrazolim(MTT)assay was used to determine the anti-tumor activity of sulfated RSGP on HepG2 and MCF-7 cell lines.Results The optimized modification conditions of RSGP were sulfating RSGP with cholorosulfonic acid-pyridine in the ratio of 1:6 for 4.5 hours at 60 ℃.The MTT assay results showed that the sulfated RSGP could inhibit the growth and proliferation of human liver cancer HepG2 cell line and human mammary cancer MCF-7 cell line in concentrationdepending manner.Conclusion It is feasible for sulfating modification of RSGP with cholorosulfonic acid-pyridine,and sulfating modification can enhance the anti-tumor activitv of RSGP.
2.Clinical analysis of capsular perforation in transurethral resection of prostate
Zhilin DENG ; Wei JIANG ; Yong CHEN ; Jun WANG
Chinese Journal of Postgraduates of Medicine 2011;34(23):21-23
Objective To investigate the causes and managements of capsular perforation in transurethral resection of prostate(TURP) for benign prostatic hyperplasia (BPH). Methods Clinical data of 893 cases who received TURP from February 2002 to February 2011 were analyzed retrospectively.Results In 893 cases, capsular perforation in 16 cases, among them, tissue covered perforation in 10 cases, dissociative perforation in 3 cases, perforation under bladder trignne in 3 cases. Six cases occurred with obviously extravasation, 10 cases with transurethral resection syndrome (TURS) or omen of TURS. Two cases were ceased operation and received TURP again after 2 weeks, 2 cases transferred to open surgery suprapubic prostatectomy, 12 cases kept up operation with lower bladder douche pressure. Two cases with postoperatire bleeding underwent open surgery hemostasis cured. All cases were discharged from hospital after recovery. Conclusions The type and extent of capsule perforation must be judged correctly,corresponding treatment must be given, prevention and treatment of TURS must be taken notice of, if capsule perforation occurs when TURP operation in progress. Less haematoma, clearview and normal anatomic structures can decrease the incidence of perforation.
3.Study on application of human cognition reliability model in human error in emergency response against the source blockage of high dose rate afterloading unit
Jun DENG ; Yaohua FAN ; Baorong YUE ; Kedao WEI ; Hao LI
Chinese Journal of Radiological Medicine and Protection 2011;31(4):404-407
Objective To put forward reasonable and feasible recommendations aiming at enhancing the application safety of afterloading unit, through studying the human reliability in the emergency response against the source blockage of afterloading unit.Methods Based on the human cognition reliability model, ten operation errors during the emergency response against the source blockage of afterloading unit were analyzed and permissible time widow of emergency response operation were determined.The human error probability was calculated with the execution time of emergency response operation obtained through simulation, observation and recording.Results The operation action, relevant permissible time window and execution time were obtained with the corresponding human error probabilities in the range 0.04 - 0.27.Conclusions The human error model in emergency response against the source blockage of afterloading unit based on HCRmodel is feasible, and provides important reference basis to reduce the occurrence of potential exposure and mitigate the consequence of potential exposure.
4.Human reliability in high dose rate afterloading radiotherapy based on FMECA
Jun DENG ; Yaohua FAN ; Baorong YUE ; Kedao WEI ; Fuli REN
Chinese Journal of Radiological Medicine and Protection 2012;32(3):314-317
Objective To put forward reasonable and feasible recommendations against the procedure with relative high risk during the high dose rate (HDR) afterloading radiotherapy,so as to enhance its clinical application safety,through studying the human reliability in the process of carrying out the HDR afterloading radiotherapy.Methods Basic data were collected by on-site investigation and process analysis as well as expert evaluation.Failure mode,effect and criticality analysis (FMECA) employed to study the human reliability in the execution of HDR afierloading radiotherapy.Results The FMECA model of human reliability for HDR afterloading radiotherapy was established,through which 25 procedures with relative high risk index were found,accounting for 14.1% of total 177 procedures.Conclusions FMECA method in human reliability study for HDR afterloading radiotherapy is feasible.The countermeasures are put forward to reduce the human error,so as to provide important basis for enhancing clinical application safety of HDR afterloading radiotherapy.
5.Implementation of performance metrics in clinical trial data management.
Ya-zhong DENG ; Hong-wei WANG ; Hai-jun FU
Acta Pharmaceutica Sinica 2015;50(11):1488-1492
There is no a systemic performance metrics for clinical data management. While the CDMC in China starts to develop the quality metrics for clinical data management, it is essential to think over the performance and pursue metrics implementation of clinical data management in China. This article provides the basic concept, development and implementation of the performance metric in clinical data management.
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Clinical Trials as Topic
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standards
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Data Collection
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standards
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Information Storage and Retrieval
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6.Changes in expression of sodium channel?subunit mRNA in dorsal root ganglion in a rat model of neuropathic pain
Ying-Wei WANG ; Jin-Jun BIAN ; Xiao-Ming DENG ;
Chinese Journal of Anesthesiology 1994;0(06):-
Objective To examine the changes in the expression of voltage-gated sodium channel?subunit mRNA in the dorsal root ganglion(DRG)and the role it plays in the neuropathie pain.Methods Thirty- two male SD rats weighing 250-400g were randomly divided into 2 groups:groupⅠneuropathic pain(SNL,n= 20)and groupⅡsham operation(n=12).Neuropathic pain was produced by ligation of right sciatic nerve according to Seltzer.Paw withdrawal latency to noxious thermal(PWHL)and mechanical(PWML)stimulation were measured before(baseline)and 1,2,3,5,8,11,14,28 day after sciatic nerve ligation(SNL).DRG at L_(4,5) was isolated on the 14th day after SNL in 8 SNL and 4 sham-operated animals for determination of sodium channel?subunit mRNA expression(by in-situ hybridization).Results PWHL and PWML were significantly decreased on the 2nd-28th day after SNL as compared to the baseline in SNL group.There was no significant difference in?_1 subunit mRNA expression between the 2 groups.The?_2 subunit mRNA expression in DRG was hardly detectable.The?_3 subunit mRNA expression in DRG on the operated side was significantly higher in SNL group than in sham-operation group.Conclusion The up-regulation of sodium channel?_3 subunit mRNA expression in DRG may play an important role in neuropathic pain.
7.Evaluation of left ventricular systolic function in different layers myocardium in patients with hypertrophic cardiomyopathy by two-dimensional speckle tracking echocardiography
Jun ZHANG ; Youbin DENG ; Qiaoying TANG ; Yang LI ; Li LI ; Wei ZHOU
Chinese Journal of Ultrasonography 2015;(4):277-281
Objective To assess the systolic function in left ventricular subendocardial myocardium , midmyocardium and subepicardial myocardium in patients with hypertrophic cardiomyopathy (HCM ) by two‐dimensional speckle tracking echocardiography .Methods A total of 32 patients with HCM and 32 healthy adults were included in the study .All standard measurements were obtained from the left ventricular short‐axis views at the levels of mitral annulus ,papillary muscle and apex ,and apical four‐chamber ,two‐chamber ,and long‐axis views .Two‐dimensional speckle tracking imaging was employed .The global peak systolic radial strain ,peak systolic longitudinal strain and circumferential strain of each layer of the left ventricle were measured .Results In patients with HCM and the control group ,the peak systolic longitudinal strain and circumferential strain reduced from subendocardial myocardium to subepicardial myocardium .The global peak systolic radial strain in patients with HCM was significant lower than that in the control group . And in patients with HCM ,peak systolic longitudinal strain in subendocardial myocardium ,midmyocardium and subepicardial myocardium was significant lower than those in the control group .Conclusions Peak systolic longitudinal strain and circumferential strain are reduced from subendocardial myocardium to subepicardial myocardium .Myocardial systolic function is impaired in patients with HCM .In patients with HCM ,longitudinal strain of each layer is lower ,but circumferential strain is mainly reduced in midmyocardium and subepicardial myocardium compared with that in the control group .
8.HPLC Determination of Saponins in Zhixiao Suppositories
Rong ZHANG ; Xiuxia DENG ; Gang WEI ; Runmei LI ; Wenzhou LI ; Jun CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2009;(7):25-27
Objective: To denvolop HPLC an method for the determination of the content of three saponins in Zhixiao suppositories. Methods: Analysis carriedout was performed on a Kromasil C18 column (4.6 mm×250 mm., 5μm)at 23℃. The mobile phases were methanol and 0.2% phosphoricacid solution in a gradient elution. The flow rate was set at 1.0 mL·min-1 and the ultraviolet detector was operated at 203 nm. Results: The linear ranges of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 were 0.368~2.76μg,0.676~5.07 μg and 0.812~6.09μg,respectively. The average recoveries of the three saponins were 98.53%, 99.20% and 99.35%, respectively. The RSD (n=5) were 2.11%, 1.88% and 2.38%, respectively. Conclusion: The separation is a good and the results are accurate and reproducible. It can be used for the quality control of Zhixiao suppositories.
9.Role of hypoxia inducible factor-1α and VHL in murine endochondral ossification
Jin SHAO ; Lianfu DENG ; Jin QI ; Qi ZHOU ; Jun WANG ; Li WEI ; Jinshen WANG ; Niandong QIAN
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(11):1305-1310
Objective To investigate the role of hypoxia inducible factor-1α (HIF-1α) and von Hippel-Lindau (VHL) in murine endochondral ossification. Methods The knockout of HIF-1α or VHL gene in murine osteoblasts was accomplished by conditional knockout technique at 4th, 8th and 12th week, and the differences between wild-type group and knock-out group in endochondral ossification were detected by HE staining, micro-CT scanning, trabecular bone area measurement, calcium content measurement, tetracycline fluorescence labeling, Real-time PCR and Western blotting. Results After knockout of HIF-1α gene in osteoblasts, the expression of vascular endothelial growth factor ( VEGF) reduced, the rate of new bone formation stepped down, the content of calcium became less, and the trabecular bone volume decreased (P <0.05) . After knockout of VHL gene in osteoblasts, the expression of VEGF increased, the rate of new bone formation stepped up, the content of calcium became more, and the trabecular bone volume was promoted (P < 0.001). Conclusion During murine endochondral ossification, VHL/HIF-1α signal pathway promotes angiogenesis through the stimulation of VEGF expression, which subsequently accelerates osteogenesis.
10.Effective expression and immunogenicity analysis of HIV-1 HXB2 subtype Tat protein deleted the cysteine-rich region in E. coli
Lu CHEN ; Songhua DENG ; Jie CAO ; Jun HE ; Qiuli CHEN ; Shaohua JIANG ; Wenting LIAO ; Wei PAN
Chinese Journal of Microbiology and Immunology 2008;28(5):404-410
Objective Deleting the cysteine-rich region (22-37 amino acids)of HIV-1 HXB2 Tat protein(whole length is 101 amino acids) to improve its stability and expression level in E.coli and to analyze the immunogenicity of Tat protein without the cystein-rich region [Tat(△C)protein]. Methods Tat DNA deleted the cysteine-rich region (64-111 nucleotides), named as Tat(△C)DNA, was obtained in vitro by PCR and cloned into pET-32a vector. pET-32a-Tat(△C)plasmid and the pET-32a-Tat plasmid were established and transformed into E.coli BL21(DE3) strains respectively to express and purify the protein. Three rabbits were vaccinated with pET-32a-Tat(△C)protein, then testify the reactivity of sera from rabbits by ELISA and Western blot. Results The dense of the purified pET-32a-Tat(△C)protein was 7.12 mg/ml,which was greatly more than pET-32a-Tat protein(1.50 mg/ml). Dimer of pET-32a-Tat protein can be observed just after the protein purification and stored at 25℃ and 4℃ for 7 days, but dimer of pET-32a-Tat(△C)protein was not formed at the same condition. Experimental rabbits were immunized with pET-32a-Tat(△C)protein and produced high titre of anti-pET-32a-Tat(△C)serum(1∶320 000), the antibody can react specifically with Tat(△C)protein, Tat protein (1-101 AA)and synthetic Tat(1-86 AA) protein. Deletion mutation of the cysteine-rich region of Tat protein was first performed in the study. Conclusion The expression level in E.coli and the stability of Tat protein deleted the cysteine-rich region can be increased greatly, and the protein remains good immunogenicity. The results may provide a novel antigen for further development of HIV-1 Tat vaccine.