2.Construction and sequence analysis of the recombinant plasmid affecting gene ATX translation by RNA interference
Ying-Hai XIE ; Bin LIU ; Ai-Ming WANG ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(12):-
Objective To construct the recombinant plasmid carrying shRNA to ATX and analyze the nucle- ic acid sequence for further searching new gene therapy method of tumor.Methods Two DNA sequences containing short hairpin structure were designed and synthesized.The complement form was obtained by annealing and inserted into vector Psilcncer2.1-U6 neo,and the recombinant plasmid was transformed into DH5a strain.Finally the plasmid identified by restriction enzyme was used for sequence analysis.Results The recombinant Psileneer2.1-U6 neo car- rying shRNA to ATX had been constructed and the aim sequence had been obtained.Conclusion The construction of the recombinant plasmid carrying shRNA to ATX lays the basis for the study of its inhibitive effect on tumor.
3.THE DETECTION OF FOOT-AND-MOUTH DISEASE VIRUS IN ANIMAL TISSUE BY RT-PCR TECHNIQUE
Gao-Ming LOU ; Wei-Xian DU ; Ao-Bin YANG ; Xiu-Rong ZHOU ; Ming-Qian XIE ;
Microbiology 1992;0(04):-
A set of primers amplified the VP1 gene of foot-and-mouth disease vims (FMDV) was designed and synthesized. A reverse transcription-polymerase chain reaction (RT-PCR) technique detected the RNA of FMDV was established after selecting the best purification method, reagents and reaction conditions. Samples of fresh milk, lymph node, spinal cord, vesicular skin, milk powder, cotton swab, mouse and meat in daughter-house were detected by RT-PCR, positive rates were41.4% (24/58), 13.33% (2/15), 20% (1/5), 100% (1/1), 100% (1/1), 37.5% (12/32), 100% (2/2) and 10% - 70%, respectively. However, positive rate of cockroach detected by RT-PCR was 0. The results showed that the established FMDV RT-PCR technique provided a more sensitive, specific and reliable method for diagnosis and epizootic study of the foot-and-mouth disease.
4.Experimental study of suspicious sensitization component screening on Huachansu injection.
Ying XIAO ; Yu-Bin ZHAO ; Yan-Ming XIE
China Journal of Chinese Materia Medica 2012;37(18):2824-2827
OBJECTIVETo explore a new experimental method for screening of allergens in post-market traditional Chinese medicine injections by confirming allergens in Huachansu injection.
METHODFirst of all, the serum of patients allergic to Huachansu injection were collected, at the same time, the dubious allergen was conjugated to bovine serum albumin (BSA) by EDC coupling procedure to form complete antigen (BNP-BSA), which makes it possible to reproduce the allergic reaction of Huachansu injection in vitro. The histamine liberation ratio, the level of TNF-alpha and Histamine released from RBL-2H3 mast cell were detected; the above data were compared with those obtained in vivo.
RESULTThe difference of the histamine liberation ratio, the levels of TNF-alpha and histamine of the resibufogenin-BSA group, group of patients allergic to Huachansu injection were not significant compared with those of normal control group. However, there were significant difference in those data among the cinobufagin-BSA group, the blank control and normal control group (P<0.05).
CONCLUSIONThe allergen in the serum collected from patients allergic to Huachansu injection is resibufogenin.
Allergens ; adverse effects ; immunology ; Amphibian Venoms ; adverse effects ; immunology ; Animals ; Anura ; Bufanolides ; adverse effects ; immunology ; Drug Hypersensitivity ; immunology ; Histamine Release ; Humans ; Mast Cells ; immunology ; Medicine, Chinese Traditional
5.Post-marketing clinical safety assessment of Shenmai injection based on active monitoring and passive monitoring in large data background.
Lian-xin WANG ; Yan-ming XIE ; Qing-hua AI ; Nian-bin SONG
China Journal of Chinese Materia Medica 2015;40(24):4752-4756
This paper adopted a series of related analysis methods to comprehensively analyze post-marketing clinical safety data of Shenmai injection from 4,220 cases of SRS and 32,358 cases of multicenter, prospective, registered hospital centralized monitoring in large data background, calculated ADR incidence rate was 0.93 per 1,000, main symptoms of ADR includes chest pain, chills, skin itching, palpitations, fever, nausea, dizziness, vomiting, flushing, numbness, allergic reaction, cyanosis, rash, low back pain, and "breath", "anaphylactoid reaction" and "flush" were the safety warning signals of Shenmai injection. Primary disease for chronic pulmonary heart disease, thyroid disease, and combined with cerebral vascular disease, prior to the injection and continuous use of alprostadil, cyclic adenosine monophosphate, combined with quinolones, penicillins were suspicious influence factors of ADR of Shenmai injection, these promot the clinical safety.
Drug Combinations
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Drugs, Chinese Herbal
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adverse effects
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Humans
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Injections
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Product Surveillance, Postmarketing
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Prospective Studies
6.Analysis suspected allergic factors to shenqi fuzheng injection based on prescription sequence analysis and nested case control study.
Qing-Hua AI ; Xian-Bin ZENG ; Yan-Ming XIE ; Wei YANG
China Journal of Chinese Materia Medica 2014;39(18):3563-3566
This study based on twenty 3A grade hospital information system (HIS) database in China, adopt toprescription sequence analysis (PSA) with nested case control study (NCCS) to analysis mainly suspected allergy factor of Shenqi Fuzheng injection (Shenqi Fuzheng). Study design according to start to stop using Shenqi Fuzheng whether using dexamethasone injection will crowd divided into cases group and control group, each case matched 4 controls were selected, two groups according to the ratio of the age and sex matched well. Square test, Fisher exact test, single factor and multiple factor logistic regression were used to analyze data Condition on admission, allergic history, dosage and drug combinations were taken into account in cases of suspected allergic reactions. After analysis in two subgroups we found that the single dose (P = 0.000 2) and the combined use of matrine (P < 0.000 1, OR = 14.312, confidence interval [8.184, 25.029]) had significant effects on the suspected allergic reaction. Study on the existing HIS data and the study method based on screening suspected risk factors for allergic reaction. This study can provide guidance for Shenqi Fuzheng injection safety using in clinical practice, and it can also provides new method for the clinical safety reevaluation of post-marketing Chinese medicine injection.
Case-Control Studies
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Drugs, Chinese Herbal
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adverse effects
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Female
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Humans
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Male
7.Analysis of questionable allergic factors to kudiezi injection--nested case control study using hospital information system data.
Xing LIAO ; Xian-Bin ZENG ; Yan-Ming XIE ; Wei YANG ; Qing-Hua AI
China Journal of Chinese Materia Medica 2014;39(18):3559-3562
This study aims to assess if adverse drug reactions (ADRs) to Kudiezi injection are allergic in origin. Hospital information system (HIS) data from 18 hospitals in China were used to carry out a nested case control design study. Included were patients who received dexamethasone for suspected allergic reactions after receiving Kudiezi injection. These were compared with non-allergic reaction people. Single factor logistic regression and multiple factor logistic regression were used to analyze data. Condition on admission, allergic history, dosage, disease status and drug combinations were taken into account in cases of suspected allergic reactions. After analysis in two subgroups we found that the condition on solvents had a significant effect, P values were 0.005 5 and < 0.000 1 on suspected cases of allergic reactions. For the first subgroup analysis, we found using other eight injections at the same time as Kudiezi injection could be risk factors in suspected cases of allergic reactions. For the second subgroup analysis combining using mannitol or fructose could increase risks. Based on this current research, condition on admission as well as the concomitant use of some other drugs may be the risk factors in suspected cases of allergic reactions. However, further research for verification is required. This study can provide guidance for safe clinical practice in using Kudiezi injection.
Adult
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Aged
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Case-Control Studies
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Drug-Related Side Effects and Adverse Reactions
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diagnosis
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Drugs, Chinese Herbal
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administration & dosage
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adverse effects
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therapeutic use
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Female
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Hospital Information Systems
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Humans
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Hypersensitivity
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diagnosis
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Injections
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Male
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Middle Aged
8.Clinical effect of Kudiezi injection on renal function based on propensity score.
Zhao-kang ZHANG ; Wei YANG ; Huan LIU ; Xian-bin ZENG ; Yan ZHUANG ; Yan-ming XIE
China Journal of Chinese Materia Medica 2015;40(13):2661-2667
To explore the effect of Kudiezi injection on renal function in the real world, in order to provide the basis for the clinical medication safety. Patient aged between 18-80 were selected from 18 large hospitals information system (HIS) databases established by clinical research institute for basic traditional Chinese medicine of China academy of Chinese medical sciences. The patients who were treated with Kudiezi injection (24 225 cases) were defined as the exposed group, whereas those who were not treated with Kudiezi injection (14,191 cases) were defined as the non-exposed group. The propensity score method was used to balance the confounding factors. Classic logistic regression, GBM weighted propensity score logistic regression, GBM propensity score weighted logistic regression with covariate and sensitivity analysis were adopted to study the effect of Kudiezi injection on renal function. The results showed no significant difference in the possibility in abnormality in serum creatinine (Scr) (P = 0.940, 0.679, 0.834) and urea nitrogen (BUN) (P = 0, 0.045, 0.164) between both groups. Therefore, the existing data indicated no damage of Kudiezi injection on renal function. Because this study is a retrospective study based on the real world, there may be unknown confounding factors and potential bias. Therefore, further studies shall be conducted to monitor whether Kudiezi injection causes damage on renal function, in order to ensure the clinical medication safety.
Blood Urea Nitrogen
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Drugs, Chinese Herbal
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adverse effects
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Hospital Information Systems
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Humans
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Injections
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Kidney
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drug effects
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physiology
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Kidney Function Tests
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Logistic Models
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Propensity Score
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Retrospective Studies
9.EGFR siRNA inhibits activation of astrocytes derived from rats through blockade of STAT3 phosphorylation
Hong QIAN ; Lijing LIU ; Yanchun LI ; Ming XIE ; Heng WU ; Shuangxi LIU ; Bin WU
Chinese Pharmacological Bulletin 2016;(2):216-222
Aim To observe the expression of epider-mal growth factor receptor ( EGFR) in cerebral tissues around hematomas after intracerebral hemorrhage, and explore the effects of EGFR on activation of astrocytes derived from rats and the involved mechanisms. Meth-ods The specimens of cerebral tissues around hemo-tomas after intracerebral hemorrhage undergoing hemo-tomas removal operation were collected and then divid-ed into 4 groups according to the time of intracerebral hemorrhage: <1 d, 1 ~5 d, 6 ~10 d and >10 d groups. Each group included 20 cases. At the same time, 20 dropped brain tissues distant to hemorrhage in the operative process were collected as control group. Immunohistostaining and Western blot were used to measure the expression of EGFR. After isolation and culturing, the astrocytes of rat cortex were treated with culture solution ( control group) , CNTF that was used to activate astrocytes, scramble siRNA + CNTF and EGFR siRNA +CNTF for 24h, respectively. The ex-pression of glial fibrillary acidic protein ( GFAP) mR-NA was detected through fluorescence real-time quanti-tative PCR. In addition, the protein levels of GFAP, signal transducers and activators of transcription 3 ( STAT3 ) and phosphorylated STAT3 ( p-STAT3 ) were examined using Western blot. Results With the ex-tension of intracerebral hemorrhage time, positive sig-nal index and protein expression levels of EGFR gradu-ally elevated, reached the peak on 6 ~10d, and then decreased after 10 d. There was statistical difference ( P<0. 01 ) . The expression levels of GFAP mRNA and protein as well as p-STAT3 were significantly in-creased in cells treated with CNTF alone as compared to control group ( P <0. 01 ) , whereas these effects were almost completely reversed by EGFR siRNA transfection ( P <0. 01 ) . Additionally, there was no statistical difference in STAT3 protein levels among groups ( P >0. 05 ) . Conclusions EGFR expression is upregulated in the cerebral tissues around hemotomas after intracerebral hemorrhage. Gene silence of EGFR contributes to suppressing the activation of astrocytes derived from rats, which may be involved in the block-ade of STAT3 phosphorylation.
10.Influence of protein phosphatase 5 on the lipid metabolism in mice
Jun WANG ; Bin QIU ; Ming LIU ; Chao WANG ; Weidong YONG ; Zhongwen XIE
Chinese Journal of Comparative Medicine 2016;26(8):79-84
Objective To investigate the effect of protein phosphatase 5 (PP5) on lipid metabolism in the PP5 knockout (KO) mice.Methods Male PP5 KO and wild type (WT) mice at the age of 6 weeks were used in this study. In order to study the effect of high fat diet ( HFD) feeding, the body weight was measured.The liver histology was examined by HE and oil red O staining.To further verify PP5 functions in the adipogenesis, in vitro experiment was carried out using mouse embryonic fibroblasts (MEF).Western blotting and real-time PCR were performed to quantified the expression of lipid metabolism-related genes in the liver tissues.Results Compared with the WT mice, the body weight gain was slower in the KO mice.The size of the lipid droplets was smaller and the quantity was less in the KO mouse liver tissue.In vitro study revealed that the KO mouse MEF cells showed less differentiated adipocytes with smaller lipid droplets than the WT MEF cells.This observation was further confirmed by detecting the expression of adipogenesis-related genes in the HFD liver.The markers of adipocyte differentiation, such as CD36, AP2, PPARγ2, and Glut4, were significantly decreased, while energy expenditure-related markers, such as phosphorylation of GR and expression of UCP1, were significantly increased.Conclusions Protein phosphatase 5 may play a regulatory role in the mouse lipid metabolism through regulating the de-phosphorylation of p-GR and enhancing the expression of UCP1.