Objective To evaluate the potential protective effects of erythropoietin (EPO) in combination with granulocyte-colony stimulating factor (G-CSF) on hypoxia cardiomyocytes. Methods After left ventricular cardiomyocytes were isolated from neonate rat, the cells were inoculated in a cultural atmosphere of 95%N2 and 5%CO2 for 24 h to establish hypoxia cardiomyocytes model. The protective effects of EPO and G-CSF at different concentrations were evaluated for the optimal concentration. Then the protective effects of EPO in combination with G-CSF were investigated under the optimized concentrations. The survival, apoptotic and necrotic rates of cardiomyocytes were assessed by flow cytometry. Results The mortality and ratio of apoptotic cells to total necrotic cell were higher in hypoxia cardiomyocytes than normal cells significantly (26.73% vs 5.63%,70.05% vs 37.83%, P0.05), and the combination group was much better than EPO and G-CSF alone groups (P

Objective To observe the impairment of different doses of cyclosporine A (CsA) to the rat myocardial tissue to offer scientific evidence for the long-term safe application of CsA in heart transplantation. Methods Eighty-four female Wistar rats, each weighing of (200 ± 25)g, were randomly divided into 12 groups. On days 7,14,21 after a constant peritoneal injection of CsA(0,5,10,15 mg/kg) and 1 ml physiological saline in control group, the rats were put to death, the rat myocardial tissue taken, to observe the pathologic and structural changes of the tissue cells under light microscope and electron microscope. The contents of rat myocardium tissue malondialdehyde(MDA) and superoxide dismutase(SOD) were measured;cardiomyocyte apoptosis was detected and accounted, apoptosis index(AI) was measured with the method of TUNEL. Results Small dose of CsA(5 mg/kg)had no obvious effects on cardiac tissue, in CsA groups of 10 mg/kg and 15 mg/kg, under the light microscope, there appeared edema, degeneration and necrosis of myocardium, part of cardiac myocyte had different level cavity;under the electron microscope, there appeared mitochondria damage, nucleus shrinkage and chromatic margination, part of cardiac myocyte had focus cavity. There was dilated endoplasic reticulum in the sarcoplasm. The effects of different time and dose on MDA content of rat myocardium tissue had statistical significance (F = 6.37,10.15, both P < 0.05). Interaction between time and dose existed statistical significance (F=7.14, P< 0.05). The MDA contents of CsA group of 10 mg/kg and 15 mg/kg were [(2.29 ± 0.18), (3.10 ± 0.45), (2.57± 0.37)nmol/L] and [(3.09±0.63), (3.32 ±0.52), (3.34 ± 0.29)nmol/L] on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [(1.98 ± 0.20), (2.04 ± 0.52), (1.99 ± 0.26) nmol/L, all P < 0.05], respectively. The effects of different time and dose on SOD activity of rat myocardium tissue had statistical significance(F = 8.43,11.69, both P < 0.05). Interaction between time and dose existed statistical significance(F = 9.86, P < 0.05). The SOD activity of CsA groups of 10 mg/kg and 15 mg/kg were (15.95 ± 1.00), (12.74 ± 1.31), (14.01 ± 0.81)nmol/L and (13.04 ± 1.01), (14.68 ± 0.81), (14.01 ± 0.63)nmol/L on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [(10.38 ± 0.80), (9.73 ± 0.58), (10.20 ± 0.26)nmol/L, all P < 0.05], respectively. Apoptosis nucleus appeared huffy or brown under the light microscope. The effects of different time and dose on AI of rat myocardium tissue had statistical significance (F = 10.02,20.46, both P < 0.05). Interaction between time and dose existed statistical significance (F = 15.73,P < 0.05). The AI of CsA groups of 10 mg/kg and 15 mg/kg were (6.91 ± 0.70)%, (11.10 ± 2.05)%,(19.81 ± 5.00)% and (11.02 ±2.02)%,(15.51 ± 1.31)%,(33.40±6.60)% on days 7,14,21 after a constant peritoneal injection of CsA, which were obviously higher than the control group [(4.40 ± 0.13)%, (4.60± 1.20)%, (5.20 ± 1.10), all P < 0.05] and CsA group of 5 mg/kg [(4.60 ± 0.10)%, (5.00±2.11)%, (5.43± 1.11)%, all P < 0.05], respectively. Conclusion Small dose of CsA has no obvious effects on cardiac tissue, but large dosage can induce myocyte apoptosis and damage by causing oxidative stress;after implantation, attention should be paid to cardiac impairment due to constant large dosage of CsA.

Objective:To detect the expressions of urea transporter B (UT-B) in tumor tissue of bladder cancer patients and normal urothelium tissue, and to investigate its clinical significance of expression in bladder cancer tissue.Methods: Fifty-two paraffin-embedded specimens of bladder cancer patients and 15 normal urothelium specimens were selected. The expression levels of UT-B protein were detected by immunohistochemistry method.The difference of expression of UT-B in bladder cancer tissue and normal urothelium tissue and its relationship with the clinicopathological parameters were analyzed.Results:The expression rate of UT-B protein in bladder cancer tissue was 44.2%, while it was significantly higher in control group (93.3%), and the difference between two groups was significant (P=0.001).The positive expression rates of UT-B in bladder cancer tissue were significantly decreased with the increasing of histological grades, and there were significant differences between different grade groups (P=0.010).The positive expression rate of UT-B in muscle-invasive stage (Ta+T1) was significantly lower than that in non-muscle-invasive stage (T2+T3) (P=0.014).Conclusion:The reduction or lack of UT-B expression may promote the incidence, progression and invasiveness of bladder cancer.

Patients with hypoxia pulmonary hypertension (HPH) are often accompanied by dyspnea, fatigue, and headache. With the development of the disease, the right ventricle gradually collapses and eventually leads to death. Hypoxic pulmonary vascular remodeling is an important pathological basis of HPH, and the remodeled pulmonary vessels will form permanent thickening. The mechanism of hypoxic pulmonary vascular remodeling is relatively complex. At present, there are few studies on drugs for pulmonary vascular remodeling on the market, mainly focusing on the alleviation of pulmonary vasoconstriction. It was found that hypoxia induces calcium overload in pulmonary artery smooth muscle cells (PASMCs), resulting in the proliferation of PASMCs. The main mechanisms include: ① abnormal expression of calcium pumps; ② abnormal calcium channels in the plasma membrane of pulmonary artery smooth muscle cells; ③ overexpression of calcium-sensitive receptors in cells; ④ the expression of Na⁺/Ca²⁺ exchanger type-1 was abnormal. This review summarized several mechanisms of hypoxia induced calcium overload leading to pulmonary artery remodeling, hoping to provide a new idea for the treatment of HPH.

OBJECTIVEPrevious studies showed potential role of tissue factor pathway inhibitor (TFPI) on attenuating restenosis, we investigated the effect of TFPI gene transfer on vascular smooth muscle cells (VSMCs) apoptosis.

METHODSHuman TFPI recombinant adenovirus or LacZ recombinant adenovirus or PBS were transferred to rat aortic VSMCs respectively in vitro. RT-PCR was used to detect the expression of exogenous TFPI gene. VSMCs were examined by cell counting and MTT. Apoptosis of VSMCs was detected by flow cytometry, TUNEL and electron microscope at different time after gene transfer.

RESULTSmRNA expression of TFPI was detected in VSMCs at the 3rd day after gene transfer. Cell numbers and absorbance value in Ad-TFPI group were similar as those in Ad-LacZ and PBS groups at the 1st, 3rd and 5th day but significantly lower at the 7th day (P<0.05) after gene transfer. The apoptosis rates in Ad-TFPI group tested by flow cytometry were all significant higher than those in Ad-lacZ groups at each time point. The positive rates in Ad-TFPI group determined by TUNEL were significant higher than those in Ad-LacZ groups at 3rd (10.82% +/- 1.57% vs. 3.46% +/- 0.93%), 5th and 7th (16.95% +/- 2.01% vs. 5.11% +/- 1.29%, all P<0.05) day post gene transfer. Electron microscope evidenced cell contracting, cytoplasm condensing, lightly swelled mitochondria, nucleus pyknosis and apoptotic body formation after gene transfer in Ad-TFPI group which were not shown in cells of LacZ and PBS groups.

CONCLUSIONTFPI gene transfer could induce apoptosis in rat VSMCs which might be one of the mechanisms responsible for its beneficial effect on restenosis inhibition after angioplasty.

Adenoviridae ; genetics ; Animals ; Apoptosis ; genetics ; Cells, Cultured ; Genetic Therapy ; Genetic Vectors ; Humans ; Lipoproteins ; genetics ; Muscle, Smooth, Vascular ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; Rats ; Rats, Wistar ; Thromboplastin ; antagonists & inhibitors ; Transfection

Objective To explore the relationship between the expression of Cyr61 and the chemosensitivity, apoptosis of human glioma U251 cells by decreasing Cyr61 gene expression by RNA interference. Methods One pair of DNA template coding siRNA was synthesized against U251 to reconstruct pRNA-Cyr61, which was transfected into U251 cells. The Cyr61 expression in U251 cells was transfected with pRNAT-Cyr61, and it was detected by RT-PCR and Western blot. MTT and flow cytometry (FCM) were used to observe the growth inhibiting ratio and apoptosis rate induced by cisplatin or temozolomide in U251 cells. Results RT-PCR and Western blot analyses demonstrated that pRNAT-Cyr61 could significantly inhibit the expression of Cyr61 in U251 cells (P＜0.01); MTT and FCM showed that when U251 cells were exposed to cisplatin or temozolomide, the growth inhibiting ratio and apoptosis rate of pRNAT-Cyr61 transfected cells were significantly increased compared with those of control group and negative group (P＜0.01). Conclusions pRNAT-Cyr61 can significantly inhibit the expression of Cyr61, increase chemosensitivity and apoptosis rate of U251 cells in vitro.

OBJECTIVETo evaluate the effects of erythropoietin (EPO) combined with granulocyte-colony stimulating factor (G-CSF) on left ventricular function and ventricular remodeling after acute myocardial infarction (AMI) and investigate the possible mechanism.

METHODSThe experimental design consisted of 5 groups of rats, namely the sham, myocardial infarction (MI) model, MI with EPO treatment, MI with G-CSF treatment, and MI with EPO plus G-CSF treatment groups. Apoptosis of the cardiomyocytes was detected by TUNEL staining, and HE staining, Masson trichrome staining, scarlatinum staining, and VIII agent staining were used to evaluate the survival, scar collagen deposition, and angiogenic effects. The cardiac structure and function of the rats after the treatments were assessed by echocardiography and hemodynamic examination.

RESULTSEchocardiography indicated that LVEF and FS were improved in all the intervention groups 7 days after MI, and the rats in EPO plus G-CSF treatment group showed the most obvious reduction of LVESD and LVESV (P<0.01). On day 28 after MI, all the intervention groups showed improvements in LVEF, FS, LVESD, LVEDD, LVESV and LVEDV, which were especially obvious in the combined treatment group; the interventions, especially the combined treatment, also resulted in decreased LVEDP and increased LVSP and +dP/dtmax. On day 1 after MI, the number of apoptotic cells was significantly greater in the MI model group than in EPO and G-CSF groups, and was the fewest in the combined treatment group (P<0.01). On day 28, the number of new vessels increased and the scar and collagen deposition reduced in the EPO and G-CSF groups, and these changes were more obvious in the combined treatment group.

CONCLUSIONSEPO combined with G-CSF can prevent left ventricular remodeling and improve cardiac systolic and diastolic functions by inhibiting cardiomyocyte apoptosis, reducing tissue collagen deposition and inducing neovascularisation.

Animals ; Drug Therapy, Combination ; Erythropoietin ; therapeutic use ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Myocardial Infarction ; drug therapy ; physiopathology ; Rats ; Rats, Wistar ; Ventricular Function, Left ; physiology ; Ventricular Remodeling ; drug effects

OBJECTIVETo study the effects of angiotensin II receptor blockers (ARB), losartan and irbesartan, on blood pressure and serum uric acid (SUA) level in mild to moderate essential hypertensive patients complicating hyperuricaemia.

METHODSA total of 351 eligible patients were recruited in this multi-center, randomized, double-blind parallel clinical trial. After 1 week screening and a 2 week single-blinded placebo wash-out period, patients were randomly assigned to receive losartan 50 mg (n=76) or irbesartan 150 mg (n=175) once daily for 4 weeks, followed by a double-dose for another 4 weeks in patients whose seated DBP were >or=90 mm Hg or SBP>or=140 mm Hg at the end of 4 weeks. The SUA concentration and blood pressure were measured at baseline, 4 and 8 weeks post therapy.

RESULTSThree hundred and twenty-five patients completed the study (162 in the losartan group and 163 in the irbesartan group). Both groups were well matched for baseline clinical characteristics and demographics. SUA was significant reduced in losartan group (430.93 micromol/L vs 372.35 micromol/L, P<0.0001), but not in Irbesartan group (430.46 micromol/L vs 420.67 micromol/L, P>0.05) 8 weeks post therapy compared to baseline level. Blood pressure was significantly and equally reduced in both groups after 8 weeks treatment compared to baseline level (P<0.0001).

CONCLUSIONLosartan is an optimum choice of medication for patients with mild-to-moderate hypertension complicating hyperuricemia.

Adult ; Angiotensin II Type 1 Receptor Blockers ; therapeutic use ; Biphenyl Compounds ; therapeutic use ; Double-Blind Method ; Female ; Follow-Up Studies ; Humans ; Hypertension ; drug therapy ; metabolism ; Losartan ; therapeutic use ; Male ; Middle Aged ; Tetrazoles ; therapeutic use ; Uric Acid ; metabolism

OBJECTIVETo investigate the expressions of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h in lung of guinea pigs with bronchial asthma, and to explore the roles of them.

METHODSForty adult male guinea pigs were randomly divided into 4 groups: the control group (group A), asthmatic group ( group B), dexamethasone group (group C) and rogridone group (group D), 10 guinea pigs in each group. The asthmatic model was established by the ovalbumin challenge method. Expressions of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h mRNA in lung tissue were assayed by in situ hybridization. Expressions of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h protein were detected by immunohischemistry and by Western blot.

RESULTSIn situ hybridization showed that the expressions of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h mRNA in lung tissue were the lowest in group B and the comparison among groups showed statistical significant (all P < 0.01). Immunohistochemistry and Western blot indicated that the value of PPAR-gamma/PGC-1alpha and Nrf2/gamma-GCS-h protein in lung tissue were the lowest in group B, and expressed primarily in nucleus, the differences being statistically significant (all P < 0.01). There was positive correlation between PPAR-gamma and PGC-1. gamma-GCS-h mRNA also positively correlated between PPAR-gamma/PGC-1alpha and Nrf2 in nucleus, and the expression of Nrf2 was also positively correlated with PPAR-gamma/ PGC-1alpha.

CONCLUSIONIn acute asthmatic models induced by ovalbumin, the expressions of PPAR-alpha/PGC-1alpha and Nrf2/gamma-GCS-h were decreased, and PPARgamma/PGC-1alpha could up-regulate the expressions of Nrf2/gamma-GCS-h to increase the antioxidant defense of tissues, thus being implicated that PPARgamma/PGC-1alpha might play important roles in the pathogenesis and prevention of asthma.

Animals ; Asthma ; chemically induced ; physiopathology ; Glutamate-Cysteine Ligase ; genetics ; metabolism ; Guinea Pigs ; Lung ; metabolism ; Male ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Ovalbumin ; PPAR gamma ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism

Objective To help the medical student to master the principle and operation of magnetic resonance imaging instrument.Methods The imaging principle,structure and functions of HT-3DNMR-25 magnetic resonance imaging instrument were described,and its application to experiment teaching was introduced including one-dimension imaging,two-dimension imaging,free induction decay signal observation and etc.Results The instrument facilitated the student to master technical principle,imaging method as well as parameters setup and regulation,and could be a supplement to clinical practical teaching.Conclusion The instrument plays an irreplaceable role in teaching,and thus is worthy promoting in colleges.