1.Rat model of exercise-induced myocardial ischemia injury
Fenglin PENG ; Lili HUANG ; Yanju GUO
Chinese Journal of Tissue Engineering Research 2016;20(40):6007-6013
BACKGROUND:No uniform standard for constructing the animal model of exercise-induced myocardial ischemia injury results in the incomparability among research results and impedes the development of sport medicine especial y in the cardiovascular field;thereby, it is imperative to reach an agreement in constructing criteria.
OBJECTIVE:To explore the method of establishing the rat model of myocardial ischemia induced by running.
METHODS:Total y 96 female Sprague-Dawley rats were randomly divided into rest control group, isoprenaline group and 10 exercise groups (1-and 3-time moderate-intensity exercise groups, 1-, 2-and 3-week moderate-intensity exercise groups, 1-and 3-time high-intensity exercise groups, 1-, 2-and 3-week high-intensity exercise groups). After exhaustive exercise, myocardium was col ected for morphological observation by hematoxylin-eosin staining, serum levels of myocardial enzymes and cardiac troponin I were detected, and the expressions of Bcl-2 and Bax were detected by real-time PCR.
RESULTS AND CONCLUSION:(1) Hematoxylin-eosin staining showed that the damage degree was more severe with the time of exercise, and the high-intensity exercise groups were more severe than those in the moderate-intensity exercise groups. (2) The activity of serum glutamic-oxaloacetic transaminase and lactic dehydrogenase was significantly increased after 1-week moderate-intensity exhaustive exercise (P<0.05 or P<0.01). From the beginning of the 3-time high-intensity exhaustive exercise, the activity of glutamic-oxaloacetic transaminase and lactic dehydrogenase was significantly increased (P<0.05 or P<0.01). (3) Cardiac troponin I content change trend was basical y the same as glutamic-oxaloacetic transaminase and lactic dehydrogenase changes, but cardiac troponin I content in the moderate-intensity exhaustive exercise groups was significantly higher than that in the rest control group until 2 weeks. The Bcl-2/Bax ratios in al exercise groups were significantly lower than that in the rest control group (P<0.05 or P<0.01);those in the 1-and 3-time high-intensity exercise groups were significantly higher than in the isoprenaline group (P<0.05 or P<0.01);and those in moderate-intensity groups were higher than in the isoprenaline group (P<0.05 or P<0.01). (4) In conclusion, 2-week high-intensity and 3-week moderate-intensity exhaustive exercise can induce myocardial ischemia injury, and pathological analysis, serum levels of myocardial enzymes and cardiac troponin I can be used as the evaluation indexes, while apoptosis regulation genes just as the reference index.
2.Accuracy of remifentanil target-controlled infusion system in children
Liguo HU ; Cai FANG ; Fenglin GUO ; Xiaohui HUANG ; Shantang ZHANG
Chinese Journal of Anesthesiology 2011;31(3):324-326
Objective To evaluate the accuracy of remifentanil target-controlled infusion (TCI) system in children.Methods Thirty ASA Ⅰ patients, aged 3-12 yr, weighing 10-40 kg, scheduled for elective ear-nosethroat or urological surgery, were randomly divided into 2 groups with 15 patients in each group:2 ng/ml remifentanil group (group Ⅰ) and 4 ng/ml remifentanil group (group Ⅱ). Anesthesia was induced with iv injection of propofol 2 mg/kg and TCI of remifentanil. Remifentanil was administered with a specific TCI system incorporating the pharmacokinetic parameters of Minto.The target plasma concentrations of remifentanil were set at 2 or 4 ng/ml. Tracheal intubation was facilitated with vecuronium 0.1 mg/kg after the children lost consciousness. The children were mechanically ventilated.Anesthesia was maintained with TCI of remifentanil, iv infusion of propofol and intermittent iv boluses of vecuronium. The target plasma concentration of remifentanil remained unchanged and bispectral index value was maintained at 45-65 or auditory evoked potentials index value < 30 by adusting the infusion rate of propofol.Arterial blood samples were taken at 5, 10, 20, 30, 40, 50 and 60 min after TCI remifentanil was stared for determination of blood remifentanil concentrations by high performance liquid chromatography. Median prediction performance error (MDPE),median absolute performance error (MDAPE) and wobble of remifentanil TCI system were calculated. Results The measured concentrations of remifentanil were significantly higher than the target plasma concentrations in both groups (P<0.05). The MDPE, MDAPE and wobble were 20.0% , 30.0% and 25.0% respectively in group Ⅰ , and 17.5%, 17.5% and 12.5% respectively in group Ⅱ . TheMDAPE and wobble were significantly decreased in group Ⅱ compared with group Ⅰ(P<0.05).Conclusion When remifentanil is administered using a specific TCI system incorporating the pharmacokinetic parameters of Minto in children of 3-12 years old, the accuracy is not high.
3.Accuracy of remifentanil target-controlled infusion system in children
Liguo HU ; Cai FANG ; Fenglin GUO ; Xiaohui HUANG ; Shantang ZHANG
Chinese Journal of Anesthesiology 2014;34(z1):21-24
Objective To evaluate the accuracy of remifentanil target-controlled infusion (TCI) system in children.Methods Thirty ASA Ⅰ patients (aged 3-12 years and weighing 10-40 kg) scheduled for elective ear-nose-throat or urological surgery were randomly divided into two groups (n =15 each):2 ng/ml remifentanil group (group Ⅰ) and 4 ng/ml remifentanil group (group Ⅱ).Anesthesia was induced with intravenous injection of propofol 2 mg/kg and TCI of remifentanil.Remifentanil was administered with a specific TCI system incorporating the pharmacokinetic parameters of Minto.The target plasma concentration of remifentanil was set at 2 or 4 ng/ml.Tracheal intubation was facilitated with vecuronium 0.1 mg/kg after the children lost consciousness.The children were mechanically ventilated.Anesthesia was maintained with TCI of remifentanil,intravenous infusion of propofol and intermittent intravenous injection of boluses of vecuronium.The target plasma concentration of remifentanil remained unchanged and bispectraI index value was maintained at 45-65 or auditory evoked potentials index value < 30 through adjusting the infusion rate of propofol.Arterial blood samples were taken at 5,10,20,30,40,50 and 60 minutes after TCI of remifentanil was started for determination of blood remifentanil concentrations by high performance liquid chromatography.Median prediction performance error (MDPE),median absolute performance error (MDAPE) and wobble of remifentanil TCI system were calculated.Results The measured concentrations of remifentanil were significantly higher than the target plasma concentrations jn both groups (P < 0.05).The MDPE,MDAPE and wobble were 20.0%,30.0% and 25.0% respectively in group Ⅰ,and 17.5%,17.5% and 12.5% respectively in group Ⅱ.The MDAPE and wobble were significantly decreased in group Ⅱ compared with group Ⅰ (P < 0.05).Conclusion When remifentanil is administered using a specific TCI system incorporating the pharmacokinetic parameters of Minto in children of 3-12 years old,the accuracy is not high.
4.Impact of Clinical Pharmacist Intervention on Reasonable Use Rate of Antihypertensive Drugs in Medical Orders
Fenglin XI ; Yi CAO ; Guilan LIU ; Fang ZHAO ; Li HUANG
China Pharmacist 2015;(1):122-124
Objective:To explore the impact of clinical pharmacists involved in the treatment team on the reasonable drug use rate in medical orders. Methods:Totally 782 medical records including hypertension and 1-3 kinds of hypertension associated diseases in our hospital from January 2011 to December 2011 were used as the pre-intervention group, and 1 061 ones with the same conditions from September 2012 to August 2013 were used as the post-intervention group. The patients of the two groups were divided into 8 dis-ease subgroups, clinical pharmacists performed timely intervention on the medical orders in the post-intervention group, and the rational drug use rate between the two groups was compared. Results:In hypertension, hypertension associated coronary heart disease, hyper-tension with heart failure and hypertension with diabetes subgroups,the difference in the rational drug use proportion before and after the intervention was statistically significant(P<0. 01), in hypertension with coronary heart disease and heart failure, hypertension associ-ated coronary heart disease and diabetes, hypertension with heart failure and diabetes subgroup, the difference in the rational drug use proportion before and after the intervention was statistically significant(P<0. 05), while in hypertension with coronary heart disease, heart failure and diabetes subgroups, the difference was not statistically significant(P>0. 05). After the medical records in the 8 sub-groups were aggregated, the difference in the rational drug use proportion before and after the intervention was statistically significant( P<0. 01). Conclusion:Clinical pharmacists should involve in the clinical treatment to intervene irrational drug use,which can signifi-cantly improve the medical record rationality, avoid the health risks as much as possible and ensure the medication safety and treatment effectiveness for patients.
5.Identification of side population cells like cancer stem cell in human pancreatic cancer cell line SW1990
Fenglin HUANG ; Shineng ZHANG ; Yijun HUANG ; Shuli WEI ; Wa ZHONG ; Haijun ZUO ; Xiaohong ZHUANG
Chinese Journal of Pancreatology 2008;08(6):372-375
Objective To isolate and identify side population (SP) cells like cancer stem cell from human pancreatic cancer cell line SW1990, for the purpose of further evaluation of their biological characteristics. Methods Cell suspension was stained with Hoechst 33342 and PI. Then SP cells were analyzed in the fluorescence activated cell sorter. Cell growth viability was measured by MTT. Stem cell marker CD133 was determined by flow cytometry. Cloning forming efficiency was determined by cloning plating. Expression of ABCG2 protein was detected by Western blot analysis. Results The proportion of SP cells was 2.7%, however it could be completely blocked by verapamil. 9 days later, the value of A492 of SP cells was 2.1, the cloning forming efficiency was (38.7 ± 6.8) % , the positive rate of CD133 was 69.63%, which were significantly higher than cells 0. 5, ( 15.5 ± 2.8)%, 16.71% of corresponding non-SP( P <0.05). The expression of ABCG2 in SP cells was significantly higher than that in non-SP cells. Conclusions SP cells existed in human pancreatic cancer cells SW1990.
6.Survivin-2B induces apoptosis of human breast cancer cells
Wenfeng ZHANG ; Jun JIA ; Fenglin WU ; Han SHEN ; Hongwei SHAO ; Shulin HUANG
Chinese Journal of Pathophysiology 2014;(6):982-987
AIM:To explore the role of survivin-2B in the process of tumor cell apoptosis .METHODS:The survivin-2B gene was cloned into pcDNA3.1 vector and the recombinant plasmid pcDNA3.1-survivin-2B was obtained.Hu-man breast cancer MCF7 cells were transfected with pcDNA3.1 and pcDNA3.1-survivin-2B using Lipofectamine 2000.The cell cycle was determined by propidium iodide staining , and the apoptosis was detected by annexin V/7-AAD staining 48 h after transfection.Meanwhile, tatal RNA was extrated and multiplex polymerase chain reaction based on GenomeLab GeXP Genetic Analysis System was performed to detect the expression of 21 tumor-related genes .RESULTS: Flow cytometry analysis indicated that over-expression of survivin-2B promoted the apoptosis and cell cycle arrest of MCF 7 cells.Compared with control group , totally 10 differential expressed genes were related to the over-expressed survivin-2B, among which 2 were up-regulated and 8 were down-regulated. The expression of aldehyde dehydrogenase 4 family member A1 (ALDH4A1) was 48%down-regulated, and the expression of protein regulator of cytokinesis 1 (PRC1) was 1.08 folds up-regulated.CONCLUSION:Survivin-2B induces the expression changes of some tumor-related genes, which results in the apoptosis and G 2/M arrest of MCF7 cells.
7.Specific TCRαβgene modification promotes anti-tumor reactivity of T cells
Fenglin WU ; Wenfeng ZHANG ; Mian HE ; Nuan YANG ; Han SHEN ; Huaben BO ; Hongwei SHAO ; Shulin HUANG
Chinese Journal of Immunology 2014;(7):901-908
Objective:To investigate effects of tumor specific TCR gene Vα12.2-Vβ7.1 modification on recognition of tumor antigen and activation of anti-tumor reactivity of T cells.Methods: T cells were transduced using recombinant Ad 5F35-TRAV-TRBV adenovirus ,and multiplicity of infection was optimized.Specific lysis of T cells was evaluated by calcein release assay.The frequency of apoptotic cells in target cells was detected by Annexin V /PI double-labeled FACS.The expression of FasL on T cells was analyzed by FACS.The secretion of cytokine IFN-γand IL-2 of T cells was determined by ELISA assays.Results: The highest tranduce efficiency was obtained at MOI 100 by recombinant Ad5F35-TRAV-TRBV adenovirus.The frequency of TCRVα12+Vβ7+cells reached above 25%3 days after transduction.TCR gene modification enhanced the ability of T cells to lyse HLA-A2+AFP+target cells(P<0.001), the ability of T cells to induce HepG-2 apoptosis(P<0.001),and expression of FasL on T cells(P<0.001).TCR gene modification also enhanced T cells to secret IFN-γafter coculture with antigen positive tumor cells ( P<0.001 ).Conclusion: Specific TCR gene modification by recombinant adeno virus effectively promotes T cells to recognize antigen positive tumor cell and exert anti -tumor reac-tivity.
8.Comparative analysis of conventional ultrasonography and contrast-enhanced ultrasound in diagnosis of renal artery stenosis
Fenglin CHEN ; Yuhua HUANG ; Yali GAO ; Yanbin HU ; Na TAO ; Hong ZHANG ; Wei ZHANG ; Nan XIA
Chinese Journal of Primary Medicine and Pharmacy 2010;17(16):2205-2206
Objective To compare contrast-enhanced ultrasound and conventional ultrasound in determining artery stenosis of varying degrees of accuracy. Methods Using conventional ultrasound and contrast-enhanced ultrasound renal artery stenosis was diagnosed.And the renal artery diameter stenosis were measured the extent to which digital subtraction angiography (DSA) as the standard diagnostic accuracy of two methods to determine the rate. Results 50 patients were diagnosed as renal artery stenosis with DSA,21 patients stenosis rate 30%-49%,23 patients stenosis rate 50%-75%,6 patients stenosis rate >75%.diagnose accordance rate 78%(38/50)with conventional color doppler and 92% (46/50) with contrast-enchanced ultrasound.The difference was statistically significant (P<0.05=. ConclusionUsing ultrasound imaging of the renal artery stenosis to determine the degree had higher accuracy than the conventional ultrasonic testing method,was suitable for clinical application.
9.Surgical repair of complete atrioventricular septal defect in 86 patients
Li XIE ; Can HUANG ; Tianli ZHAO ; Jinfu YANG ; Zhongshi WU ; Yifeng YANG ; Fenglin SONG
Journal of Chinese Physician 2016;18(8):1152-1155
Objective To investigate the early surgical outcomes of 86 patients with complete atrioventricular septal defect.Methods Between January 2007 and December 2014,consecutive 86 cases received surgical repair in our department.There were 44 male patients,and 42 female patients.Two-patch repair was performed in 69 cases,and modified single-patch repair in 17 cases.The mean age,height,and weight at the time of operation were (32.3 ± 46.5)months with a range from 1 month to 17 years,(82.1 ±27.6) cm with a range from 53 to 165 cm,and (10.8 ± 8.7) kg with a range from 4.1 to 43 kg,respectively.Rastelli A type was found in 67 cases,B type in 15 cases,and C type in 4 cases.Down's syndrome was complicated in 6 cases.Preoperative mild regurgitation of common atrioventricular valve was shown in 32 cases,moderate regurgitation in 38 cases,and moderate to severe regurgitation in 16 cases.Mild pulmonary hypertension was observed in 15 cases,moderate in 54 cases,and severe in 17 cases.Results After operation,all patients were sent into intensive care units (ICU).The mean duration mechanical ventilation,ICU stay,and hospitalization were (30.9 ± 47.7) h with a range from 2.5 to 244 h,(87.7 ± 76.8) h with a range from 14 to 306 h,and (16.4 ±9.2)d with a a range from 6 to 50 d,respectively.We encountered 4 operatively mortalities (4.7%),including 3 in two-patch repair group,and 1 in modified single-patch repair group.The cause of death was mitral regurgitation.Conclusions Modified single-patch and two-patch technique have a satisfied early outcomes.
10.Development of a novel assay for cytotoxicity test by Calcein-AM release
Changli TAO ; Zhechun DING ; Wenjing GUO ; Fenglin WU ; Hongwei SHAO ; Shulin HUANG
Chinese Journal of Immunology 2014;(10):1364-1368
Objective:To develop and optimize a novel assay for determination of cytotoxicity based Calcein -AM release.Methods:The target cells stained by Calcein-AM dye,then effectors and targets were incubated at E/T ratios from 30∶1-1∶1 for 4 h at 37℃,and the supernatant of reactions were detected by Fluorescence-Measurement to analyze specific cytotoxity.Results:The optimal excitation and emission wave lengths of Calcein were 485 nm and 515 nm.Dilutions of target cells stained by Calcein-AM had a linear relationship with measured fluorescence values.The Calcein-AM dye used to stain the living cells was shown to have a low spontaneous leakage rate-less than 15% in 4 hours at 37℃.Cytotoxicity activity of CIK showed a significant and positive correlation with E/T ratio when incubated at 4 h.Conclusion:The developed cytotoxicity test by Calcein-AM release is accurate and can avoid the application of radioactive reagents.