1.Improvement of cervical heterotopic heart transplantation in mice using Tail-Cuff technique
Chinese Journal of Tissue Engineering Research 2010;14(31):5883-5886
BACKGROUND: The mouse model of cervical heart transplantation is an ideal medical research tool for study of transplant-induced ischemia/reperfusion injury and immunological rejection.However,technical problems have limited the widespread use of mouse cervical vascularized heart transplantation.OBJECTIVE: To improve the cervical heterotopic heart transplantation in mice using the tail-cuff technique.METHODS: Isogeneic transplantation was performed from Balb/c to BALB/c mice,and allogeneic transplantation from C57BL/6 to BALB/c mice.The right common carotid artery and the external jugular vein of the recipient were equipped with a tail cuff made from 24 G and 22 G intravenous catheter,and everted over the cuff,and then connected with the aorta and the pulonary artery of donor heart,respectively.RESULTS AND CONCLUSION: A total of 36 transplants for formal experiment,12 for isogeneic transplantation,and 24 for allogeneic transplantation,were performed with a surgical successful rate of 100%.The total surgical procedure was(49.6±7.4)minutes and total ischemic time of the grafts was(28.8±4.2)minutes.In particular,the average time for vascular everting and for the reconnection of both vessels was obviously shortened.This improved tail-cuff technique shows its superiority,and can serve as an ideal method for establishing cervical heterotopic heart transplantation model in mice.
2.Advances in the study of the immunoregulatory mechanism of acupuncture-moxibustion treatment for rheumatoid arthritis
Yinping WANG ; Xiaozheng DU ; Zongquan ZHANG
Shanghai Journal of Acupuncture and Moxibustion 2017;36(1):108-112
This article summarizes recent years’ advances in the study of the immunoregulatory mechanism of acupuncture-moxibustion treatment for rheumatoid arthritis (RA) and shows that RA occurrence involves a complex process in which immunocytes, cytokines, Immunoreactive substances and neuro-endocrino-immune network participate together. The article proposes that the future study should utilize the newest achievements in modern medical development and conduct a thorough and systematic investigation from multiple disciplines, levels and angles to promote the study of the immunoregulatory mechanism of acupuncture and moxibustion in RA treatment and make traditional acupuncture-moxibustion therapy still play its unique advantages in the RA field.
3.Effect of Lovastatin preconditioning on cardiac myocyte apoptosis in ischemic and reperfused rat hearts
Xiaojie ZHANG ; Zongquan SUN ; Xinling DU
Chinese Journal of Thoracic and Cardiovascular Surgery 1995;0(05):-
Objective To study the effects and mechanisms of Lovastatin preconditioning on cardiac myocyte apoptosis in acute ischemic and reperfused (I/R) rat hearts. Methods 24 Sprague-Dawley male rats were randomly divided into three groups: control group (C); Lovastatin group (L) treated with lovastatin 15 mg/kg once a day for two weeks; Lovastatin and L-NAME group (N) treated with the same dose of Lovastatin with L group and L-NAME 30 mg/kg once a day for two weeks. Rat heart models of I/R were established with coronary occlusion 30 mintues and reperfusion 30 mintues of the left anterior descending artery after two-week administration. Expression of Bcl-2,Bax protein and myocardium apoptosis were investigated in every group. Results There was no change in blood lipin, expression of Bax protein was increased (P
4.Inhibitory effect of methylene chloride-induced on cardiac allograft rejection in a murine heart transplantation model
Songlin ZHANG ; Zongquan SUN ; Li YU ; Yuan ZHOU
Chinese Journal of Organ Transplantation 2012;33(4):241-245
Objective To investigate the role of induction of carbon monoxide (CO) with methylene chloride (MC) in recipients in ameliorating allograft rejection and prolonging allograft survival and to explore the possible mechanisms in a murine heart transplantation model.Methods Inbred male C57BL/6 mice were used as donors and inbred male Balb/c mice as recipients respectively to establish cervical heterotopic heart transplantation model.The experiments were divided into 3 groups.Recipients were treated with MC (100 mg/kg,per os) day 1 prior to transplantation to day 6 posttransplantation (group MC1w) or to day 13 posttransplantation (group MC2w),or treated with isovolumic olive oil day 1 prior to transplantation to cardiac arrest of allograft (group Tx).The serum TNF-α and IL-10 proteins,TNF-α and IL-10 mRNA,and Foxp3 mRNA and protein in cardiac grafts were measured respectively.The intercellular adhesion molecule-1 (ICAM-1) and Caspase-3 protein in cardiac grafts,and the histopathologic changes of cardiac grafts were also observed.Results Serum COHb levels in untreated mice were (0.85 ± 0.28)%.After MC application,serum COHb peaked within 3 h in recipients (5.24 ± 0,45)% (P<0.01 ).The median survival time of cardiac grafts in group MC1w(12.1 days) and group MC2w( 19.4 days) was longer than that in group Tx (6.3 days) (P <0.01). As compared with group Tx,induction of CO in group MC1w and group MC2w down-regulated significantly the levels of serum TNF-α (P<0.01 ) and TNF-α mRNA (P<0.01) of cardiac grafts and spleen in recipient mice,inhibited the protein expression of ICAM-1 (P<0.01) and Caspase-3 (P<0.01) of cardiac grafts,and inhibited,especially in group MC2w,the proliferation of lymphocytes and monocytes infiltration in cardiac grafts.There was no significant difference in serum IL-10 and Foxp3 mRNA and protein in cardiac grafts and spleen of recipients among the groups (P>0.05).Conclusion Induction of CM in recipients could relieve cardiac allograft rejection and prolong cardiac allograft survival via its anti-inflammation and anti-apoptotic effects,not via up-regulation of Foxp3 in recipient mice.
5.Effects of HIF-1α expression regulated by Tet-on gone expression system on hepatoma cell proliferation and cell cycle in vitro
Ronglong XIA ; Zongquan XU ; Ruiqin ZHANG ; Xiaoping CHEN
Chinese Journal of General Surgery 2010;25(10):834-837
Objective To investigate the effects of HIF-1α expression regulated by Tet-on gene expression system on cell proliferation and cell cycle of hepatoma cells in vitro. Methods The change of human hepatocellular carcinoma cell lines HepG2 cell cycle and cell proliferation was measured after HIF-1 α expression of HepG2 in vitro was regulated by Tet-on expression system. Results Amplified products were confirmed as the cDNA of HIF-1α by DNA sequencing, and pTRE-HIF-1α obtained by edonuclease digestion,capable of expression in HepG2 Tet-on cells. After being incubated under different concentrations of doxycycline for 48 h, MTT assays showed that up-regulation of HIF-1α expression increased HepG2 cell proliferation activities. The cell index of S and G2/M phase was significantly higher and that of G0/G1 phase reduced with the increasing concentrations of doxycycline. The mRNA expression of Cyclin A increased with the increasing concentrations of doxycycline ( P < 0. 001 ), CyclinD1 and CyclinE did not change ( P >0. 05). Conclusion HIF-1 α gene promotes cell proliferation and cell cycle of hcpatoma cells in vitro, and this effects increased with the increasing of HIF-1α expression possibly through influencing the expression of CyclinA.
6.The increase of carbon monoxide in recipients ameliorates isehemia/reperfusio.injury in a murine heart transplantation model
Songlin ZHANG ; Zongquan SUN ; Jiane FENG ; Long WU ; Li YU
Chinese Journal of Organ Transplantation 2010;31(3):157-161
Objective To examine whether the increase of carbon monoxide (CO) induced by oral methylene chloride (MC) administration in recipients before heart transplantation would protect heart grafts against isehemia/reperfusion (I/R) injury associated with transplantation and to explore the possible mechanism.Methods Inbred male Balb/c mice were used as donors and recipients to establish cervical heart transplantation model Recipients were treated with either MC (100 mg/kg or 500 mg/kg,per os)(group MC 100 mg,n=10;group MC 500 mg,n=12) or olive oil(0.15 ml,per os.group olive,n=10) 3 h prior to anesthesia.Age-matched norwlal mice served as controls (group N,n=5).The serum COHb and the CO content of myocardial tissue were measured at 0,1,3,6,12,24 h after oral MC administration.Half of recipients were killed at 3 and 24h after transplantation for senum or cardiac graft samples.The serum cTnI levels,the mRNA levels of TNF-α,IL-10,Bcl-2,Bax.the protein levels of NF-κB and the ultrastructures of myocardium were examined.Results As tompared with group olive.the serum COHb and tissue CO were increased significantly and peaked within 3 h in group MC 100 mg and group MC 500 mg.The serum cTnI levels in group MC 100 mg and group MC 500 mg were significantly decreased (P<0. 01 ), especially in group MC 500 mg. The increase of CO in recipients of group MC100 mg and group MC 500 mg significantly inhibited the proinflammatory gene expression of TNF-α mRNA and the pro-apoptotic gene expression of Bax mRNA (P<0. 01), and increased the anti-apoptotic gene expression of Bcl-2 mRNA (P<0. 01), but did not increase the anti-inflammatory gene expression of IL-10 mRNA (P>0. 05) in the heart grafts. As compared with group N, the myocardial NF-κB activation was increased significantly in group olive,group MC 100 mg and group MC 500 mg (P<0. 01 ), but there was no significant difference among the later three groups (P>0. 05). The myocardial ultrastructure was also alleviated significantly in group MC 100 mg and group MC 500 mg as compared with group N. Conclusion The increase of CO induced by MC in recipients suppresses pro-inflammatory and pro-apoptotic gene expression and efficiently ameliorates transplant-induced heart I/R injury. The possible mechanism does not seem to be associated with down-regulation of the NF-κB signaling pathway.
7.Science of Meridians, Collaterals and Acupoints--Exploration on teaching method of meridian syndromes.
Yinping WANG ; Zongquan ZHANG ; Wenlin WANG ; Limin YUAN
Chinese Acupuncture & Moxibustion 2015;35(4):385-387
Meridian syndromes are the required basic knowledge for mastering Science of Meridians, Collaterals and Acupoints but have not brought the adequate attention on the teaching program. The writers discovered' that the content of this section occupied a decisive role for developing the students' clinical thinking ability and, stimulating their interests to learn classical TCM theories. It's necessary to enhance the importance on meridian syndromes during teaching program. The teaching program was discussed in three aspects, named workshop pattern, competitive pattern and multimedia pattern. This teaching method may improve students' interests in the study on classical TCM theories, deepen the understanding on knowledge and motivate students' learning autonomy so that the teaching quality can be improved.
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8.Impact of NF-kappaB Activation on TNF-α and IL-1β Expression in Myocardial Ischemia/Reperfusion Injury
Tucheng SUN ; Xionggang JIANG ; Kailun ZHANG ; Zongquan SUN
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2009;38(6):752-755,759
Objective To study the impact of NF-kappaB activation on TNF-a and IL-1β expression in myocardial ischemia/reperfusion(I/R) injury.Methods Sixty-five Sprague-Dawley rats were randomly divided into three groups:sham,n=5;I/R:30 min of myocardial ischemia followed by 0,15,30,60,120.240 min of reperfusion,n=5 per subgroup;I/R+PDTC:PDTC(15 mg/kg)was given before ischemia,and the time points were the same as those in I/R group.TNF-a and IL-1β mRNA expression was detected by RT-PCR,activity of NF-KB was measured by electrophoretic mobility shift assay (EMSA),and MDA level in myocardium was assayed by TBA method.Results The expression of TNF-a and IL-1β was increased before reperfusion,reached their peak at the time point of reperfusion 30.60 min respectively.and remained high level at the 2nd h after reperfusion.NF-KB was activated 15 min after reperfusion,reached its peak at the first h after reperfusion.In I/R+PDTC group,NF-κB activation was blocked by PDTC.As compared with I/R group,the expression levels of TNF-a and IL-1β were decreased to varying degrees at each time point.and the content of MDA was also reduced in I/R+PDTC group.Conclusion NF-KB activation could play a pivotabrole in the expression of cytokine.Inhibition of NF-κB signal pathway might be a potential therapeutic strategy in reperfusion injury.
9.The effect of Tet-on gene regulated HIF-1?expression on hepatoma cell apoptosis in vitro
Zongquan XU ; Xiaoping CHEN ; Wanguang ZHANG ; Changhai LI ; Gaopeng LI
Chinese Journal of General Surgery 2000;0(11):-
Objective To investigate the effects of Tet-on gene regulated HIF-1?expression on cell apoptosis of hepatoma cells in vitro. Methods The apoptosis of hepatocellular carcinoma cell lines HePG2 was measured after HIF-1?expression was activated in HePG2 in vitro by Tet-on expression system. Results The amplified products were confirmed as the cDNA of HIF-1?by DNA sequencing, and obtained pTHE-HIF-1?was verified by endonuclease digestion. The HePG2Tet-on cells express HIF-1?constantly. After being incubated under different concentrations of doxycycline for 48 h, HIF-1?gene could inhibit HePG2 cell apoptosis. The apoptosis rates were 59. 6%、50. 9%、38. 1%、30. 5%、23. 9%、18. 3% respectively when the concentrations of doxycycline were 0?g/ml,0. 02?g/ml,0. 2?g/ml、1?g/ml、2?g/ml,5?g/ml (P
10.Mitral valve reconstruction and replacement for moderate to severe ischemic mitral regurgitation: comparison of midterm outcome and complications
Xuefeng QIU ; Nianguo DONG ; Zongquan SUN ; Shiliang XIAO ; Kailun ZHANG ; Xinling DU ; Xionggang JIANG ; Wei SU
Chinese Journal of Thoracic and Cardiovascular Surgery 2010;26(2):95-98
Objective To compare the results of mitral valve reconstruction and replacement as treatments for moderate to severe ischemic mitral regurgitation(IMR), and report the mid-term outcome. Methods From June 2002 to May 2008, 83 pa-tients with moderate IMR(35 cases) and severe IMR (48 cases) underwent coronary artery bypass grafting(CABG) combined with mitral valvuloplasty (MVP) (n = 43) or mitral valve replacement (MVR) (n = 40). There were 49 males and 34 females with a mean age of (59.3±7.5) years(51 -77years). The procedures of MVP included annuloplasty with a Dacron or autologous per-icardium ring in 21cases, commissural annuloplasty in 9, quadrangular resection of the posterior leaflet in 9 and using St. Jude mitral annuloplasty ring in 4. In the cases underwent MVR, 28 patients received mechanical prostheses and 12 received biopros-theses. Results 30-day mortality rate was 2.3% for MVP and 5.0% for MVR (P >0.05). The 30-day complication rate was similar for the 2 groups but mechanical ventilation time was longer for MVR patients. Mild MR ocurred in 6 patients with MVP (P <0.05). Sevonty-six patients were followed by outpatient department visit or telephone for (20.2 ± 4.9) months (3 - 60 months). During the follow-up period, 7 patients with MVP had mild insufficiency but free off etber complications. All the valve prothesis functioned well. However, 3 cases had thromboembolic complications and 7 late deaths were recorded in MVR group. Five-year complication-free survival rate was 90% for MVP group and 61% for MVR. Conclusion MVP resulted in excellent durability and provided significant mid-term survival benefit over MVR. MVP should be the first choice for patients with chronic IMR.