3.Role of sonic hedgehog signaling pathway in spinal neurons in morphine tolerance in mice
Junli YAO ; Su LIU ; You LYU ; Peiyu CAO ; Longjian YAN ; Hui SU ; Gongjian LIU
Chinese Journal of Anesthesiology 2017;37(2):175-179
Objective To evaluate the role of sonic hedgehog (SHH) signaling pathway in spinal neurons in morphine tolerance (MT) in mice.Methods Pathogen-free healthy female Kunming mice,weighing 20-25 g,aged 8-10 weeks,were used in the study.MT was induced with morphine 10 mg/kg injected subcutaneously twice a day for 7 consecutive days.The experiment was performed in two parts.Experiment Ⅰ Forty-eight mice were randomly assigned into 2 groups:control group (group C,n =8) and MT group (group M,n=40).The thermal pain threshold (TPT) was measured at 1 day before morphine injection and 1,3,5,7 and 14 days after the end of injection.Eight mice in each group were sacrificed at 2 h after measurement of TPT at each time point after the end of injection in group M or at 2 h after the last measurement of TPT in group C,and the lumbar segment (L4-6) of the spinal cord was removed.Experiment Ⅱ Forty-eight mice were randomly assigned into 6 groups (n=8 each):SHH inhibitor cyclopamine plus MT group (group CP+M),cyclopamine solvent plus MT group (group D1 +M),SHH agonist SAG plus MT group (group SAG+M),SAG solvent plus MT group (group D2+M),MT plus cyclopamine group (group M+CP) and morphine plus cyelopamine solvent group (group M+D1).At 15 min before morphine injection,cyclopamine 10 mg/kg was injected subcutaneously in group CP+M,and SAG 5 mg/kg was injected subcutaneously in group SAG+M.Cyclopamine 10 mg/kg was injected subcutaneously once a day during the 1-3 days after the end of morphine injection in group M+CP.The TPT was measured before injection of morphine,at 30 min after the first injection of morphine every day and at 1-3 days after the end of morphine injection.The animals were sacrificed at 2 h after the last measurement of TPT,and the lumbar segment (L4-6) of the spinal cord was removed for determination of the expression of SHH signaling pathway-related proteins SHH,ptch1,smo,gli1 and gli3 using Western blot.Results Experiment Ⅰ Compared with group C,the TPT was significantly decreased at 1 and 3 days after the end of morphine injection (P<0.05),no significant change was found in TPT at 5-14 days after the end of morphine injection (P>0.05),and the expression of SHH,smo and glil at 1-5 days after the end of morphine injection,of ptchl at 1 and 3 days after the end of morphine injection and of gli3 at 7 days after the end of morphine injection was up-regulated in group M (P<0.05).Experiment Ⅱ Compared with group D1+M,the TPT was significantly increased,the expression of SHH,ptchl,smo and glil was down-regulated,and gli3 expression was up-regulated in group C P+M (P<0.05).Compared with group D2+M,the TPT was significantly decreased,the expression of SHH,ptch1,smo and glil was up-regulated,and gli3 expression was down-regulated in group SAG+M (P<0.05).There was no significant difference in the parameters mentioned above between group M+CP and group M+D1 (P>0.05).The TPT was significantly lower on 3rd-7th days after beginning of morphine injection and 1-3 days after the end of morphine injection than at 30 min after the first injection of morphine in group CP+M (P<0.05).Conclusion The mechanism underlying the development of MT is partially related to activation of SHH signaling pathway in spinal neurons of mice,however,the maintenance mechanism has no marked relationship with it.
4.Clinical research for patients with diabetes undergoing off-pump coronary artery bypass grafting by endoscopic saphenous vein harvesting
Xitao ZHANG ; Jie CAO ; Yan LIU ; Song GU ; Jun YAN ; Yue XIN ; Pixiong SU
Chinese Journal of Thoracic and Cardiovascular Surgery 2016;32(3):159-161
Objective To compare clinical effects and complications of patients with diabetes in off-pump coronary artery bypass grafting(OPCAB) between endoscopic saphenous vein harvesting(EVH) and conventional saphenous vein harvesting(CVH).Methods Sequence comparison analysis the clinical data of 339 patients with DM who underwent OPCAB in our department from Nov 2011 to Nov 2014.269 cases by EVH,70 cases by CVH.Observing two groups of patients with deprecated rate of SVG,intraoperative SVG blood flow and the value of PI,lower limb wound complications such as incision infection,poor healing,lower limb local hematoma and pain.SVG patency rate of part patients was follow-up by CT coronary angiography.Results To compare the two groups of patients by EVH and CVH,the perioperative death was 8 cases in EVH group (2.4%),2 cases in C VH group (2.9%).The deprecated SVG of patients was 3.9% vs 2.9%.The blood flow was (17.36 ±11.24) ml/min vs(17.11 ± 8.37) ml/min,PI was 2.78 ± 2.37 vs 2.22 ± 2.17.The incision infection was 0 vs 4.4%,poor healing was 0.9% vs 8.8%.The lower limb local hematoma was 5.7% vs 1.5%.The visual pain analogue scale(VAS) was 0.53 ± 1.71 vs 1.26 ± 2.13 3 days after operation.The numbness of lower limb was 9.7% vs 22.1%.The Edema of the legs was 8.5% vs 19.1% 7 days after operation.60 cases were follow-up by CT coronary angiography,the SVG patency rate was91.4% vs 94.6% 1 year after operation,83.3% vs 86.1% 2 years after operation,72.2% vs 73.7% 3 years after operation respectively.Conclusion EVH technology for SVG in the patients combined DM has good clinical result,the recent patency rate of SVG is perfect,postoperative limb complications is decreased by EVH.
5.Risk factors and prognosis for hemorrhagic transformation caused by intravenous thrombolysis treatment with recombinant tissue plasminogen activator treatment in acute cerebral infarction
Min SU ; Weixin YANG ; Wanhua WANG ; Yongjun CAO ; Taosheng LU ; Xuanfei JIANG ; Yan KONG ; Chunfeng LIU
Chinese Journal of Neurology 2011;44(11):754-758
Objective To investigate risk factors and prognosis of hemorrhagic transformation(HT)in acute cerebral infarction patients treated by intravenous thrombolysis with recombinant tissue plasminogen activator(rt-PA).Methods All 128 patients with acute cerebral infarction were treated with intravenous rtPA within 6 hours from stroke onset.The clinic records and laboratory datas of pre-and post-treatment were statistically analyzed between HT group and non-HT group to find potential risk factors to HT and contributors of prognosis.Results HT occurred in 29 patients(22.66%),including 16 patients with symptomatic ICH(12.50%)and 2 patients died(6.90% of HT).Logistic regression analysis showed that history of atrial fibrillation(OR =1.293,95% CI 1.224-1.589,P =0.001),CT density changes with mass effect or edema(OR =2.452,95% CI 1.132-3.309,P =0.034),diastolic blood pressure ≥ 100 mm Hg before thrombolytic therapy(1 mm Hg =0.133 kPa,OR =9.265,95% CI 1.435-59.836,P =0.019),blood glucose ≥ 11.1 mmol/L(OR =3.037,95% CI 0.252-57.593,P =0.047),NIHSS score > 15 points (OR =8.752,95% CI 1.035-30.285,P =0.023)and thrombolysis time > 3 h(OR =98.74,95% CI 5.067-186.120,P =0.002)are independent risk factors for HT; among these factors,baseline blood glucose(OR =3.265,95 % CI 0.435-59.863,P =0.045),NIHSS score(OR =10.453,95 % CI 5.647-38.185,P =0.003)and thrombolysis time(OR =2.541,95% CI 1.098-51.086,P =0.017)also are prediction factors of the prognosis of HT.Conclusion Risk factors associated with HT are diastolic blood pressure before thrombolysis,glucose level,degree of neurological deficits,CT early changes,atrial fibrillation and thrombolytic time.Glucose level,neurological deficits and thrombolysis time affects the prognosis of patients.
6.Prognosis comparison of vascular in situ and bridge vessel percutaneous coronary intervention in patients with recurrent angina after coronary artery bypass grafting
Haijian CHEN ; Ni MO ; Xiulong ZHU ; Yong CAO ; Zhibao WEI ; Yan CHEN ; Guozhu SU ; Handong WU
Clinical Medicine of China 2017;33(9):802-806
Objective To compare the prognosis of vascular in situ and bridge vessel percutaneous coronary intervention ( PCI) therapy strategies in patients with recurrent angina after coronary artery bypass grafting ( CABG) . Methods A total of one hundred and two patients with recurrent angina after CABG from January 2008 to January 2016 were involved in this study and were divided into two groups according to interventional therapy strategy:74 patients in the vascular in situ PCI group ( in situ group,74 cases) and 28 patients for bridge vessel PCI group ( bridge vessel group,28 cases) . The patients have been followed up for (33. 6± 10. 2) months. The major adverse cardiovascular events ( MACE) of the two groups were recorded, including non?fatal acute myocardial infarction ( AMI) ,target vessel revascularization ( TVR) and cardiac death, and multivariate logistic regression analysis was used to analyze the related factors of MACE. Results Compared with the bridge vessel group,the non?MACE survival rate,non?AMI survival rate and non?TVR survival rate of the in situ group were significantly increased ( ( 71. 6% ( 53/74 ) vs. 57. 1% ( 16/28 ) , 93. 2% ( 69/74 ) vs. 82. 1% (23/28),81. 1% (60/74) vs. 67. 9% (19/28) ),the differences were statistically significant (χ2=8. 141,4. 219,5. 436, P<0. 05) . Multivariable logistic regression analysis showed that age of bridge ( OR=1. 023,95%CI 1. 005-1. 026,P=0. 019) ,diabetes mellitus ( OR=2. 386,95%CI 1. 425-3. 991,P=0. 003) and bridge vessel PCI (OR=1. 884,95%CI 1. 093-3. 220,P=0. 025) were factors that affect the clinical prognosis in patients with recurrent angina pectoris after CABG. Conclusion The clinical prognosis of the in situ PCI is better than bridge vascular PCI in patients with recurrent angina after CABG,while the age of bridge, diabetes mellitus, vascular interventional treatment are factors for the effect of interventional therapy patients prognosis. The clinical prognosis is much better in native vessel PCI than that of bridge vessel PCI in patients with recurrent angina after CABG. The age of bridge,diabetes mellitus and bridge vessel PCI are the factors that affect the clinical prognosis in the patients.
7.Comparison of native vessel percutaneous coronary intervention and optimal drug therapy in patients with recurrent angina pectoris after coronary artery bypass grafting
Haijian CHEN ; Ni MO ; Xiulong ZHU ; Yong CAO ; Zhibao WEI ; Yan CHEN ; Guozhu SU
Chinese Journal of Postgraduates of Medicine 2017;40(9):807-810
Objective To compare the efficacy of native vessel percutaneous coronary intervention (NV-PCI) and optimal drug therapy (ODT) in patients with recurrent angina after coronary artery bypass grafting (CABG). Methods The clinical data of 142 recurrent angina pectoris after CABG patients who had underwent coronary angiography were retrospectively analyzed. Among the patients, 70 cases were treated with NV-PCI (NV-PCI group), and 72 cases were treated with ODT (ODT group). The incidence of major adverse coronary events (MACE) and left ventricular ejection fraction (LVEF) were compared between 2 groups. Results All patients were followed up for at least 1 years. There were no statistical differences in the number of bypass vessels and number of occluded vessels between ODT group and NV-PCI group: (2.5 ± 0.7) branches/case vs. (2.4 ± 0.9) branches/case and (1.4 ± 0.9) branches/case vs. (1.3 ± 0.7) branches/case, P>0.05. The incidence of MACE in NV-PCI group was significantly lower than that in ODT group: 12.9% (9/70) vs. 22.2% (16/72), and the LVEF was significantly higher than that in ODT group:(63.5 ± 14.0)%vs. (57.1 ± 9.0)%, and there were statistical differences (P<0.05). Conclusions Compared with the ODT, the NV-PCI has lower incidence of MACE and higher LVEF in patients with recurrent angina pectoris after CABG.
8.Effects of low-dose of oral thephylline on sputum inflammatory cells and inflammatory mediators in patients with COPD
Yan YIN ; Liang CAO ; Hongjian SU ; Gang HOU ; Qiuyue WANG ; Jian KANG
Journal of Chinese Physician 2016;18(5):675-679
Objective To investigate the anti-inflammatory effects of low-dose and sustained release oral theophylline on the chronic obstructive pulmonary disease (COPD) patients.Methods Thirty four patients with stable COPD were randomly divided into two groups:theophylline group (n =18) was treated with slow-release theophylline (100 mg,twice daily),and placebo group (n =16) was given with placebo.Healthy non-smokers (n =12) were taken as control.The course of treatment was 12 weeks both of theophylline group and placebo group.The percentages of Neu/Leu and Mφ/Leu in sputum were detected before and after treatment and the concentrations of interleukin (IL)-17,IL-8,and tumor necrosis factor-α (TNF-et) were detected with enzyme linked immunosobent assay (ELISA).Results (1) Compared to pretreatment with theophylline group,the Neu/Leu was increased [(89 ±4.14)% vs (83.4 ±6.98)%,P <0.05] and the Mφ/Leu was decreased [(6.4 ± 4.11) % vs (12.3 ± 6.96) %,P < 0.05] in the post-treated theophylline group.No significant changes in both Neu/Leu and Mφ/Leu were observed before and after placebo-treatment (P > 0.05).(2) Compared to the control group,the concentrations of TNF-α,IL-8,and IL-17 in the sputum supernatant were significantly increased in both pretreatment and posttreatment with the theophylline or the placebo.Sputum TNF-α,IL-8,and IL-17 levels were significantly decreased in COPD patients who were given theophylline.Compared to pre-treatment with placebo group,the IL-8 and IL-17 levels were significantly increased in the post-treated placebo group (P <0.01).There was no significant change in TNF~ level between before and after treatment with the placebo.(3) The concentrations of IL-17,IL-8,and TNF-α in the sputum supernatant were positively correlated with the Neu/Leu counts (r =0.471,0.652,0.466,respectively,all P <0.01),negatively correlated with the forced expiratory volume in one second (FEV1 %) (r =-0.516,-0.652,-0.496,respectively,all P < 0.01).Conclusions Low-dose and sustained-release oral theophylline was efficient in improving airway inflammatory cells and inflammatory mediators,which plays an anti-inflammatory effect.
9.Clinical observation on the treatment of low-dose oral theophylline in patents with COPD
Yan YIN ; Hongjian SU ; Liang CAO ; Gang HOU ; Qiuyue WANG ; Jian KANG
Journal of Chinese Physician 2016;18(6):837-841
Objective To investigate the effects of low-dose,sustained release oral theophylline on the chronic obstructive pulmonary disease (COPD) patient.Methods Fifty-six patients with stable COPD were randomly divided into two group:theophylline group (n =35) that was treated with slow-release theophylline(100 mg,twice daily),and control group (n =21) that was given with placebo.A series of parameters including lung function,quality of life scores,body mass index,airflow obstruction,dyspnea,and exercise capacity index (BODE) score,exercise tolerance,exacerbations,satisfaction with treatments and adverse effects were tested before and 12 weeks after the treatments.Results Forty two patients completed the study,25 cases in the slow-release theophylline group,and 17 cases in the placebo group.The differences of two groups before the treatment were not prominent except the age (P > 0.05).After treated with slow-release theophylline,the forced expiratory volume in one second (FEV1),forced vital capacity (FVC) and the symptom score were slightly increased,but there were no statistically significant differences (P >0.05).After theophylline therapy,the quality of life score,including activity ability score,disease activity score and total score,and BODE index score were significantly decreased(P <0.05),but 6 minutes walk test (6 MWT) differences were no significant (P >0.05).The differences in pulmonary function test,the quality of life score,BODE index score and 6 minutes walk test were no significant between before and after the treatment with the placebo (P > 0.05).Compared to the cases who treated with the placebo group,the patients in slow-release theophylline group reduced the frequencies of acute exacerbation of chronic obstructive pulmonary disease (AECOPD) (3/25 vs 7/17,x2 =4.748,P <0.05),and increased the efficacy satisfaction (Z =-2.579,P < 0.05).Slightly adverse reaction was observed in 3 cases in slow-release theophylline group,but it could relieve by oneself,and not affect the common treatment.There was no adverse reaction in the placebo group.Conclusions Low dose,sustained release oral theophylline was efficient in improvement of the quality of life scores and BODE index score.
10.Construction of TK Gene-deleted PRV SH StrainContaining a Single LoxP Site
Min-Xiu WANG ; Xin-Ming SU ; Chun-Mei YU ; Rui-Bing CAO ; Pu-Yan CHEN ;
China Biotechnology 2006;0(10):-
Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.