1.Validation of zebrafish embryo toxicity evaluation method
Jia CHANG ; Liang LU ; Yan CHANG
Chinese Journal of Pharmacology and Toxicology 2014;(2):290-295
OBJECTIVE In this study,nine different levels of known teratogenic embryos toxic com-pounds (aceta minophen,methi mazole,indo metacin,methotrexate,ascorbic acid,isoniazid,penicillin G,saccharin ,5-fluorouracil )were used to validate the established evaluation method of zebrafish e mbryo develop mental toxicity in our laboratory via zebrafish e mbryonic develop ment toxicity tests. METHODS By water bath method,zebrafish embryos of post-fertilization 2 h (2 hpf)were exposed to different concentrations of compounds.Observe and record the nu mber of malformations and dead e mbryo in 24,48,72,144 hpf.After three parallel experiments,get EC50 ,LC50 through software and calculate the teratogenic indices (TI =LC50 /EC50 )of the positive drug and recorded it as teratogens.If there is no teratogenic index recorded it as non-teratogens.Co mpare the results with the existing animal experiments and clinical trials to obtain the sensitivity and specificity.RESULTS Based on the results, exposure to aceta minophen (TI =2.07),methi mazole (TI =2.91 ),indo methacin (TI =1 .67),5-flu-orouracil (TI =8.31 ),methotrexate (TI =1 .31 ),showed various levels of teratogenic effects,and the corresponding TI′s were calculated.Methi mazole appeared more obvious skeletal deformities;indo m-ethacin toxicity is mainly reflected in abnormal heart,blood circulation and other endpoints;5-FU at lower doses that result in higher mortality rates,but also took place over the lacking of pig mentation;aceta min-ophen for the heart,liver and other target organ toxicity,pericardial cyst;methotrexate appears signifi-cant so mites,skeletal deformity.Since the IT′s for ascorbic acid,isoniazid,penicillin G,saccharin can-not be calculated,these compounds are considered to show negative results,indicating that the data fro m the nine compounds in zebrafish test are consistent with traditional animal experiments and clinical information,the sensitivity of 100% and specificity of 100%.CONCLUSION Though validation test,it has been de monstrated that the established zebrafish embryos develop mental toxicity evaluation method can be applied to early screening of drug toxicity.
2.Inhibition of ovarian cancer growth by small interfering RNA targeting X-linked inhibitor of apoptosis gene
Jia-Jia MA ; Bi-Liang CHEN ; Xiao-Yan XIN ;
Chinese Journal of Obstetrics and Gynecology 2001;0(02):-
Objective To construct an RNA interference vector to down-regulate X-linked inhibitor of apoptosis(XIAP)gene and study the RNA interference effect on the cell cycle and growth of ovarian cancer.Methods Oligonucleotides of 64 base pairs for hairpin RNA targeting XIAP were designed, chemically synthesized,annealed,and cloned into the pSUPER vector.After identification by restriction digestion,the correct vectors were transiently transfected into SKOV3 cells,a human ovarian cancer cell line.The XIAP mRNA was detected by RT-PCR.The proteins were detected by western blot and indirect immunofluorescence staining.Flow cytometry(FCM)analysis and methyl thiazolyl tetrazolium(MTT)assay method were applied to measure cell cycle,cell growth and sensitiveness to cisplatin.Results SKOV3 cells had a high level expression of XIAP.The vector of RNA interference,which can interfere with XIAP gene was successfully constructed.After transient transfection,the expression of XIAP protein was significantly decreased in SKOV3 cells and the value of relative density was 3584?124,2138?65,1973?80 and 110 ?12,respectively(P=0.0334).At the same time,the expression of XIAP mRNA was decreased accordingly and the value of relative density was 6674?274,4532?107,2322?57 and 1864?78, respectively(P=0.0127).The FCM results showed that,the vector could increase the number of cells in G_1 phase compared with parent cells and compared with the cells transfected with pSUPER(P
3.DNA repair protein degradation mediated by helicase-like transcription factor in radiation-induced apoptotic cells
Yingbao ZHU ; Yun HAN ; Yan SUN ; Li LIANG ; Tingzhen JIA
Chinese Journal of Radiological Medicine and Protection 2010;30(5):510-512
Objective To study the effects of helicase-like transcription factor (HLTF)transfection on DNA repair protein level in radiation-induced apoptotic cells.Methods Human lung carcinoma A549 cells were cultured and transfected with FLAG-tagged wild type HLTF (wild type HLTF transfection group),RING structure domain (ubiquitin conjugating region) mutatation HLTF expressing plasmid (mutant transfection group),empty plasmid (congtrol group) respectively.And the other cells were used as mock transfection group.All cells were irradiated with 15 Gy of 60Co γ-rays to induce apoptosis.Western blotting was used to detect the protein levels of the DNA repair proteins HRAD17 and HRAD52 in the transfected cells.Results The levels of HRAD17 and HRAD52 in the wild type HLTF transfection group was significantly lower than that of the control group.There was no significant difference in HRAD17 and HRAD52 levels between the mock transfection group and ubiquity in conjugating region mutation group.complexes of HLTF and HRAD17 and HRAD52 could be found in the irradiation-induced cells.Conclusions HLTF mediates the degradation of HRAD17 and HRAD52 in the irradiation-induced apoptotic cells possibly by the interaction of the protein complex causing ubiquitination of the repair proteins.
4.Case report: peritonsillar abscess with deep cervical space and mediastinal infection.
Hai-yan WANG ; Hai-ying JIA ; Hong LIANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2011;46(10):865-866
Adult
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Humans
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Male
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Mediastinitis
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complications
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Neck
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Peritonsillar Abscess
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complications
5.Expression and significance of interleukin-6 in children with chronic rhinosinusitis
Bing WANG ; Yan SHU ; Jia LIANG ; Hongbing YAO
Chongqing Medicine 2016;(1):19-20,23
Objective To evaluate the expression of interleukin-6 (IL-6) on peripheral blood and adenoid in children with chronic rhinosinusitis (CRS) and adenoidal hypertrophy ,to investigate the mechanism of IL-6 on the occurrence and development of CRS .Methods A total of 216 cases of hypertrophic adenoids were collected ,including 95 CRS children with hypertrophic adenoid (CRS group) ,70 children with allergic rhinitis and hypertrophic adenoid (AR group)and 51 children with hypertrophic adenoid (control group) .The plasma IL-6 concentrations were detected by ELISA ,and the expression and distribution of IL-6 in adenoid tissue was detected by immunohistochemistry .Results The plasma IL-6 level in CRS group was (49 .8 ± 8 .3)pg/mL ,significantly higher than that of in control group (33 .6 ± 6 .9)pg/mL and in AR group (35 .9 ± 5 .6)pg/mL ,differences had statistic significance ( P< 0 .05) .However ,there was no statistically significant difference between AR group and control group .Meanwhile ,the presence of IL-6 in adenoid tissue was mainly seen in inflammation cells and fibroblasts while less in mesenchymal cells ,there were statistical differences between CRS group and control group/AR group (P< 0 .05) ,but no statistical difference between AP group and control group(P> 0 .05) .Conclusion Among children with CRS ,expression of IL-6 whether in peri-plasma or in focal tissue was higher than those without CRS ,demonstrating that IL-6 may play an important role in the pathogenesis and developemnt of CRS in chil-dren .
6.Gastroscopic, endoscopic ultrasonographic, immunohistochemical and clinicopathological features of esophageal mesenchymal tumors
Guobao JIA ; Yan ZHOU ; Liang WU ; Zhiming HUANG
Chinese Journal of Digestion 2013;33(8):507-512
Objective To explore the features of clinicopathology,gastroscopy,endoscopic ultrasonography (EUS) and immunohistochemistry of esophageal mesenchymal tumors (EMT).Methods The clinical data of 98 cases of EMT patients who underwent endoscopic biopsy,endoscopic treatment or surgery and were diagnosed according to pathological and immunohistochemistry examination were collected.The clinicopathologic characteristics,endoscopy,EUS and immunohistochemistry of these cases were retrospectively analyzed,and differentiated from other submucosal tumor.Results Among the 98 cases of EMT,there were 77 leiomyomas (LM),15 gastrointestinal stromal tumors (GIST),four lipomas,one leiomyoblastoma and one schwannoma.The age of GIST group were older than that of LM group,and most were male patients predominated in these two groups.Dysphagia was the most common symptom of EMT.Gastrointestinal bleeding was rare,which was different from other gastrointestinal mesenchymal tumors.Under endoscopy,the appearances of EMT were similar,as submucosal protuberant lesions.One esophageal GIST was extraluminal lesion.Under EUS,LM,GIST and schwannoma showed low echo images originated from the muscularis propria or muscularis mucosa.Lipomas showed high echo images originated from submucosa.Pathological examination indicated that main cell type of LM and schwannoma was spindle cell,part of GIST was epitheloid cell type.The expressions of CD117 and CD34 were high in GIST,the expressions of SMA and Desmin were high in LM.S-100 was expressed in schwannoma.Other submucosal tumors,such as esophageal cyst,esophageal tuberculosis,esophageal carcinosarcoma,and some esophageal squamous carcinoma were easily misdiagnosed as EMT.Conclusions Esophageal LM is the most common EMT,followed by GIST.EUS is helpful in EMT diagnosis,however it can not accurately distinguish GIST,LM or schwannoma.
7.Clinical Study of Niaoshitong Pill in the Treatment of Urinary Calculus with Syndrome of Qi and Damp Stagnation
Yan MO ; Liuji MO ; Feng LIANG ; Fusheng TIAN ; Jinming JIA
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(02):-
Objective To evaluate the curative effect and safety of Niaoshit on g pill in the treatment of urinary calculus.Method Multi- center randomized co ntrolled clinical trial was adopted. Three hundred and twenty cases were accepte d to the study, in which 200 cases were treated by Niaoshitong pill and 120 case s by Shilintong tablet as control. The effect of both groups was observed. Resul t 107 cases (53.5 % ) were cured, 53 cases(26.5 % ) effective, the total effe ctive rate being 80.0 % in the treatment group, and 27 cases(24.5 % ), 42 cas es (38.2 % ), and 62.7 % respectively in the control group. In a open group of 120 cases ,54 cases (45.0 % ) were cured, 44 cases (36.6 % ) were effective , the total effective rate being 81,6 % .Conclusion Niaoshitong pill can mark edly improve the clinical symptoms and exerts a strong lithagogue effect. It can promote the elimination of calculi after external blast lithotrity or ureterosc opic lithotrity, prevent the formation of 'stone street', and reduce the strictu re formed by the damage of ureter.
8.Study on the Effect of Various Polarity Fractions Extracted from Sancaofang and Its Combination on Human Lung Adencarcinoma SPC-A-1 Cells
Zhenzhong SHAO ; Xiaobin JIA ; Yan CHEN ; Liang FENG ; Feng SHI
China Pharmacy 2001;0(07):-
OBJECTIVE: To study the effect of various polarity fractions extracted from Sancaofang (SCF) and its combination on proliferation of human lung adenocarciaoma SPC-A-1 cells for bolting the most effective position of anti-tumor and suitable compatibility regimes.METHODS: Ethanol extraction and water extraction were adopted to prepare 95%,60% and 30% ethanol extract portion,water extract portion of SCF and compound decoction.The MTT assay was used to determine the effect of various polarity fractions of SCF,decoction and its combination on SPC-A-1 cells proliferation.RESULTS: IC50 of 60% ethanol extract was the smallest for SPC-A-1 cells.60% ethanol extract combined with 95% ethanol extract acts as a stimulus to anti-tumor activity significantly.CONCLUSION: The best suitable compatibility regimes were 95% ethanol extract combined with 60% ethanol extract.The liposolubility extract of SCF can be applied for anti-tumor.
9.Cathepsin L expression in plasma after acute myocardial ischemia and ischemia-reperfusion in rats.
Geng-qian ZHANG ; Zheng LIANG ; Peng YAN ; Xiao-jia ZHANG
Journal of Forensic Medicine 2014;30(4):253-256
OBJECTIVE:
To test cathepsin L as a biomarker of myocardial ischemia by examination of cathepsin L expression in plasma after myocardial ischemia and ischemia-reperfusion in rats.
METHODS:
The rat models were established and divided in acute myocardial ischemia model (myocardial ischemia 30 min, 1 h, 2 h groups), ischemia-reperfusion model (ischemia-reperfusion group), and isoflurane-pretreated ischemia-reperfusion model (isoflurane-pretreated group), respectively. Normal control group and sham-operated group were established as contrast. The contents of cathepsin L in plasma were examined by ELISA and myocardial infarction areas were measured after TTC staining.
RESULTS:
No statistical significant changes were found among the experimental groups compared with the normal control group and sham-operated group (P>0.05). The cathepsin L from the ischemia-reperfusion group increased to 2.37 times compared with the normal control group (P<0.05). The cathepsin L and myocardium infarction size of isoflurane-pretreated group decreased compared with the ischemia-reperfusion group (P<0.05).
CONCLUSION
The cathepsin L in plasma is not a promising biomarker of acute myocardial ischemia. Isoflurane preconditioning can reduce the cathepsin L in plasma caused by ischemia-reperfusion injury.
Animals
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Biomarkers/blood*
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Cathepsin L/analysis*
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Isoflurane
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Myocardial Infarction/metabolism*
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Myocardial Ischemia
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Myocardial Reperfusion Injury/metabolism*
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Myocardium
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Rats
10.Research about improving flowability of powder of Chinese herbs extracts by surface modification technology.
Yan-Hong YU ; Wen-Liang LU ; Jia-Jia LI ; Changquan Calvin SUN ; Qun ZHOU
China Journal of Chinese Materia Medica 2014;39(23):4590-4595
In this study, modification technology by surface coating was used to improve the flowability of powder of Chinese herbs extracts. Seven kinds of powder of Chinese herbs extracts were coated with 1% silica nanoparticles using an under-driven Comil. The powder characteristics tester was used to evaluate the flowability of uncoated and coated powders. Surface morphology and particle size distribution of powders were compared by scanning electron microscope (SEM) and optical microscope. The powder hygroscopicity was also investigated. The results showed that, after modification, angle of repose, angle of spatula, compressibility and adhesiveness extremely decreased, and flowability index substantially increased, the powder flowability was significantly improved, especially Gegen and Dahuang extracts powders. Scanning electron microscopy images showed the distribution of silica nanoparticles on the host drug particle surfaces. There were no remarkable changes in the particle size distribution and hygroscopicity of all powders after coating. Therefore, Comilling for surface coating modification technology was an effective method to improve the flowability of Chinese herbs extracts and suggested a possible way forward to enhance the quality of Chinese drugs pharmaceutics in their study and manufacture.
Chemistry, Pharmaceutical
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methods
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Drugs, Chinese Herbal
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chemistry
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Nanoparticles
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chemistry
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Particle Size
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Powders
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chemistry
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Surface Properties