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Objective:To investigate the effect of hydroxysafflor yellow A(HSYA) preconditioning group on apoptosis induced by cold hypoxia/reoxygenation (cold H/R) injury in human renal tubular epithelial cells (HK2 cells).Methods:After digestion and passage, HK2 cell lines were divided into Sham group (control group), cold hypoxia and reoxygenation group (cold H/R group, cells cold hypoxia for 4 h, reoxygenation for 4 h), and HSYA preconditioning group (each HSYA subgroup was given different doses of HSYA 0.5 h before hypoxia, and the other operations were the same as the cold H/R group). The cell survival rate was measured by CCK-8 method.The expression of Bcl-2, Bax and Caspase-3 proteins in HK-2 cells were detected by immunocytochemistry and Western blotting.Results:(1) Compared with cold H/R group, different doses of HSYA could improve cell survival rate in different degrees, but only HSYA25 μmol/L group had the most significant effect (74.000±5.500 vs.59.000±3.800, P<0.05). (2) Immunocytochemistry semi-quantitative score: Compared with cold H/R group, the expression of Bax and Caspase-3 in HK2 cells of HSYA25 μmol/L group was significantly decreased(0(0, 1) vs. 8(6, 8), Z=2.041, P<0.05 and (3.400±0.548) vs.(7.800±1.095), t=11.000, P<0.01). The expression of Bcl-2 protein was increased significantly ((6.800±1.095) vs.(1.400±0.548), t=10.590, P<0.01). The ratio of Bcl-2/Bax increased significantly.(3)Western blot was used to detect protein: Compared with the cold H/R group, the protein levels of Bax, Cleaved-Caspase-3 and Pro-caspase-3 of HK2 cells in the HSYA25 μmol/L group were significantly decreased ((0.707±0.012) vs.(0.968±0.117), (0.480±0.009)vs.(0.735±0.005), (0.992±0.008)vs.(1.197±0.005), all P<0.01). The expression of Bcl-2 protein was significantly increased, and the ratio of Bcl-2/Bax was significantly increased ((0.410±0.009) vs.(0.273±0.008), (0.582±0.016) vs (0.282±0.080), all P<0.01). The experimental results were consistent with the immunocytochemistry. Conclusion:HSYA can effectively reduce the damage of HK2 cells after cold hypoxia and reoxygenation.
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Objective To observe the effect of HIF-1a/iNOS signaling pathway on myocardial ischemia-reperfusion injury in rat heart transplantation and the protective mechanism of N-acetylcysteine (NAC) on donor heart after cardiac transplantation in rats.Methods Eighty healthy male Lewis rats were randomly divided into 3 groups,the control group (0.3 ml saline was infused via inferior vena cava 30 min before donor harvest or implantation),NAC donor pretreatment group [NAC (30 mg/kg.w) was injected into the vena cava of donor rat 30 min defore donor harvest],and the NAC receptor pretreatment group (NAC 300 mg/kg.w was injected into the vena cava of the recipient rats 30 min before transplantation.The 30 min was injected into the vena cava of the recipient rats).A transplant model was established and the graft was obtained after 24 h transplantation.The expression of iNOS,HIF-1a and mRNA in cardiac muscle tissue was detected by immunohistochemistry and Real time-PCR.Results HIF-1a protein expression in graft myocardial tissue was significantly lower in NAC donor pretreatment and recipient pretreatment group compared with control group (P ＜0.05),the differences were statistically significant (2.72±0.17 vs.2.24±0.23 vs.3.14.±0.16,F=56.26,P =0.000).The iNOS protein expression in NAC donor pretreatment group,and NAC recipient pretreatment group were lower than that in the control group (1.52±0.18 vs.1.61±0.19 vs.3.30±0.18,F=232.345,P =0.000),the differences were statistically significant (P ＜ 0.05).24 h after transplantation,the differences in graft myocardial tissue HIF-1a and iNOS mRNA among the three groups were statistically significant (F=7.467,16.490,P=0.003,0.000).The expression of iNOS mRNA in the NAC receptor pretreatment group was significantly lower than that in the control group (P＜0.05).Conclusion HIF-1a/iNOS signaling pathway can regulate ischemia reperfusion injury in rat heart transplantation,and the protective effect of NAC on donor heart maybe mediated via this pathway.
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Objective:To explore the effect of sulforaphane (SFN) preconditioning on the cold myocardial ischemia-reperfusion injury (IRI) in the rats through PI3K/Akt signaling pathway.Methods:Sixty-four health male Sprague-Dawley (SD) rats were randomly divided into cold IRI group,SFN group,LY (LY294002) + cold IRI group,and LY+SFN group (n=16).The allogeneic heterotopic heart transplantation model was established by donor heart into recipient abdomen.The myocardium tissue was taken 24 h after reperfusion for the detection of histological changes using HE staining.The expression levels of Akt,p-Akt,Bax and Bcl-2 proteins were detected by immunohistochemistry and Western boltting methods.Results:The morphological results showed that the myocardium tissue damage was serious in cold IRI group and LY+cold IRI group,it was light in SFN group;the myocardium tissue damage of the rats in SFN+ LY group was ranged between cold IRI group and SFN group.Compared with IRI group,the expression levels of p-Akt protein and Bcl-2 protein in SFN group were increased (P＜0.05),and the expression level of Bax protein was decreased (P＜0.05).After treatment of blockage LY294002,compared with LY-+-cold IRI group,the expression level of p-Akt protein in LY-+-SFN group was not statistically significant (P＞0.05),the expression level of Bcl2 protein was increased (P＜0.05),the expression levels of Bax protein was decreased),and the ratio of Bcl-2/Bax was also increased (P＜0.05).Conclusion:SFN may attenuate cold IRI of heart transplantation through PI3K/Akt signaling pathway in the rats.
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Objective:To explore the effect of sulforaphane (SFN) preconditioning on the cold myocardial ischemia-reperfusion injury (IRI) in the rats through PI3K/Akt signaling pathway.Methods:Sixty-four health male Sprague-Dawley (SD) rats were randomly divided into cold IRI group,SFN group,LY (LY294002) + cold IRI group,and LY+SFN group (n=16).The allogeneic heterotopic heart transplantation model was established by donor heart into recipient abdomen.The myocardium tissue was taken 24 h after reperfusion for the detection of histological changes using HE staining.The expression levels of Akt,p-Akt,Bax and Bcl-2 proteins were detected by immunohistochemistry and Western boltting methods.Results:The morphological results showed that the myocardium tissue damage was serious in cold IRI group and LY+cold IRI group,it was light in SFN group;the myocardium tissue damage of the rats in SFN+ LY group was ranged between cold IRI group and SFN group.Compared with IRI group,the expression levels of p-Akt protein and Bcl-2 protein in SFN group were increased (P＜0.05),and the expression level of Bax protein was decreased (P＜0.05).After treatment of blockage LY294002,compared with LY-+-cold IRI group,the expression level of p-Akt protein in LY-+-SFN group was not statistically significant (P＞0.05),the expression level of Bcl2 protein was increased (P＜0.05),the expression levels of Bax protein was decreased),and the ratio of Bcl-2/Bax was also increased (P＜0.05).Conclusion:SFN may attenuate cold IRI of heart transplantation through PI3K/Akt signaling pathway in the rats.
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BACKGROUND: The indicant of successful transplanted bone marrow stem cells is that the labeled transplanted ceils can survive in the target organs and can their biological functions. Up to date, at cell level, there are several labeled methods such as enzyme-linked, 3H-TdR, fluorescent and 5-bromodexyuridine method, etc.OBJECTIVE: To observe the biological characteristics of bone marrowderived mesenchymal stem cells labeled with 5-bromodexyuridine. DESIGN: Observations in single kind of sample SETTING: Department of Thoracic Cardiac Surgery, Affiliated Hospital of North China Coal Medical College MATERIALS: The experiment was carried out in the Experimental Center of North China Coal Medical College from July, 2003 to November,2004 on six Japanese long-eared rabbits ,with the age of 3 months , of either gender and the body mass of (3.00±0.25)kg.METHODS:①Monocytes were isolated from the bone marrow with Percoll reagent of the concentration of 1 073 g/L. The mesenchymal stem cells were cultured and proliferated with Eagle's medium, which was improved by adding 10% low sugar Dulbecco's to fetal bovine serum, so that their purity could reach about 95%. Secondly, in the experimental groups, the percentage of bone marrow-derived mesenchymal stem cells labeled with 5-bromodexyuridine was detected after 24 hours, 48 hours, 72 hours and 96hours, respectively. The negative-controlled group was composed of bone marrow-derived mesenchymal stem cells non-labeled with 5-bromodexyuridine while the blank-controlled group was made by substituting phosphate buffer or normal mice serum for primary antibody.MAIN OUTCOME MEASURES: In three different groups, 200 cells of each group were detected and observed at different time. Outcomes of the immunochemical stauning and counting of bone marrow-derived mesenchymal stem cells labeled with 5-bromodexyuridine were also determined.RESULTS: The positive bone marrow-derived mesenchymal stem cells labeled with 5-bromodexyuridine were observed in all the experimental groups. The positive materials were brown, granular and dfiffusedly scattered in the nucleus while no positive cell was observed in the controlled groups. After 24 hours, 48 hours, 72 hours and 96 hours, the numbers of bone marrow-derived mesenchymal stem cells labeled with 5-bromodexyuridine were 48±2, 100±4, 173:t:2, 178±3 respectively. The labeling percentage was raised gradually with the elongation of time. 72 hours later,the labeling percentage is above 85%. The numbers in negative-controlled and blank-controlled groups were all zero.CONCLUSION: The appropriate time of bone marrow-derived mesenchymal stem cells labeled with 5-bromodexyuriding is 72 hours. The sensibility of post-labeling detection is high due to the results being observed in the low power microscope, which makes this method suitable to quantitative analysis in massive tissue. These results show that the method of bone marrow-derived mesenchymal stem cells labeled with 5-bromodexyuridine is feasible.
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BACKGROUND: A majority scholars views that polymorphism of angiotensinogen T174M gene is one of the susceptible factors of inheritance of coronary heart disease, hypertension and myocardial infarction.OBJECTIVE: To probe into the relationship between the variation of angiotensinogen T174M gene and myocardial infarction.DESIGN: Case-controlled verified experiment.SETTING: Department of Cell Biology, Biochemistry and Molecular Biology, Biological Science Faculty of North China Coal Medical College,and Department of Cardiology in Affiliated Hospital of North China Coal Medical College.PARTICIPANTS: Fifty-five cases of myocardial infarction were collected from outpatients and inpatients in Department of Cardiac Vascular Internal Medicine of Worker's Hospital Affiliated to North China Coal Medical College in Tangshan from September 2002 to September 2003, of which,29 cases were males and 26 cases females, aged (60±8) years. At the same time, 60 cases (health control) were selected from the people who received clinical physical health check (without repeated physical check), of which,32 cases were males and 28 cases females, aged (60±10) years. The cases selected had no manifestation of coronary heart disease, without history myocardial infarction or cerebral infarction in family and the participants were in the know of the research.INTERVENTIONS: Polymerase chain reaction was used to amplify the genetic sequence of No.174 DAN residue involved in No.2 exon of angiotensinogen gene. Electrophoresis was used after variated with Nco I restriction endonuclease.Analysis of restriction fragment length polymorphism was carried on angiotensinogen genotype. Simultaneously,the relevant risk factors of coronary heart disease were detected in two groups, such as blood pressure, body mass, blood lipid, fasting blood glucose, etc.MAIN OUTCOME MEASURES: ① Distribution of genotype, frequency of genotype and frequency of allele in two groups. ② Analysis on risk factors of two groups.RESULTS: Totally 115 cases of objects all accomplished the design and entered result analysis. ① Frequency of angiotensinogen genotype: in myocardial infarction group, TT 75％ (41/55), TM 18％ (10/55), MM 7％ (4/55) and in control group, TT 83％ (50/60), TM 15％ (9/60), MM 2％ (1/60). Frequency of allele of M174 and T174 were 16％ (18/110), 84％ (92/110) and 9％ (11/120), 91％ (109/120) in myocardial infarction group and control group respectively. Frequency of allele of M174 in myocardial infarction group was significantly higher than that of control group (x2=5.79,P ＜ 0.05). By the division of sex, the frequency M and T alleles of both male and female in experiment group was basically identical to control group. Angiotensinogen 174MM genotype in myocardial infarction group was significantly higher than control group (x2= 7.55, P ＜ 0.025). ② Comparison of risk factors: The percentage of smoking history in myocardial infarction group was significantly higher than control group (P = 0.006). After correction of essential risk factors of coronary heart disease, angiotensinogen 174MM gene still increased significantly the risk of myocardial infarction (Odds ratio was 3.66, P= 0.018).CONCLUSION: Angiotensinogen genotype is related to the occurrence of myocardial infarction. M allele is one of the susceptible factors of inheritance of myocardial infarction and T allele prevents from myocardial infarction. The attack of myocardial infarction is not relevant to sex, but angiotensinogen 174TM gene is one of the essential risk factors of myocardial infarction.
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Objective To evaluate the effect of left ventricular assist device(LVAD) on acute left heart failure canine model by the alteration of local AngII receptor(AT1/AT2) mRNA expression in left ventricular myocytes. Methods 15 canines were randomly divided into 3 groups: Control, hert failure(HF) and HF+LVAD groups. Acute left heart failure model was induced by occluding the left anterior descending coronary artery (LAD) or LAD+left circumflex coronary artery (LCX). The LVAD group was treated for 3 hours by pneumatic LVAD, whose output was 70?ml?kg. -1.?min. -1.. 100mg myocardium was taken from the left ventricle of each dog at 1, 2, 3 hours respectively. The expression of the local left ventricle AngII receptor (AT1/AT2) mRNA was tested by reverse transcription-polymerase chain reaction (RT-PCR). Results When compare with the control group, the expression of the AT1 mRNA in ischemic left ventricle decreased significantly in HF group after heart failure; AT2 mRNA in ischemic left ventricle increased significantly in HF group after heart failure. At 1, 2, 3 hours after LVAD, the AT1 mRNA level of the HF+LVAD group was higher than that of the HF group's, the AT2 mRNA level of the LVAD group was lower than that of the HF group's. Conclusion After heart failure, the expression of AT1 mRNA decreased, but it can be reversed by the LVAD. The expression of AT2 mRNA increased, and it can be reversed by the LVAD. The improvement of the AT1 mRNA and AT2 mRNA values in HF+LVAD group shows the improvement of left ventricular function.
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Objective To compare the clinical effects between hysteroscopic electroresection and open hysterectomy in the treatment of intrauterine benign diseases. Methods A total of 58 women with intrauterine benign received hysteroscopic electroresection (hysteroscopic group) and 60 women underwent open total hysterectomy (control group). The operating time, hospital stay, postoperative recovery and cure rates were compared between the two groups, respectively. Results The operating time was 68.4?20.6 min in control group and 55.2?19.7 min in hysteroscopic group ( t=-3.555, P =0.000). The postoperative hospital stay was 7.2?1.8 d and 4.5?1.5 d in control and hysteroscopic group, respectively ( t=-8.836, P =0.000). The time to resume regular work was 64.4?25.3 d in control group and 37.2?7.8 d in hysteroscopic group ( t=-7.835, P =0.000). The cure rates in control and hysteroscopic group were 100% (60/60) and 94.7% (54/57), respectively ( ? 2 =1.477, P =0.224). Conclusions Hysteroscopic electroresection gives good short-term therapeutic effects and quick postoperative recovery. It may replace hysterectomy in part of patients with intrauterine benign diseases.
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AIM:To evaluate the protective effect of hypothermic ventricular fibrillation without aortic-cross clamping under cardiopulmonary bypass(CPB)on canine lung.METHODS:Fourteen dogs were randomly divided into two groups.All dogs received a standardized anesthetic technique.A conditional CPB was performed in every instance.Ventricular fibrillation was induced by systemic hypothermia to 28 ℃ and pericardial cooling saline in the experimental group.A standard CPB was performed in control group.The concentration of IL-8 in serum was measured by ELISA.The expressions of NF-?B and ICAM-1 were determined by using immunohistochemical staining.RESULTS:Serum IL-8 level in experimental group was significantly lower than that in control group(P