OBJECTIVETo investigate the influence of human C-type natriuretic peptide (hCNP) on proliferation of vascular endothelial cells (HUVECs).

METHODSReconstructed pcDNA3.1 (+)/hCNP was transfected into HUVECs with polyethylenimine and its plasmid expression was examined with RT-PCR, immunohistochemistry and Western blot. MTT method was used to determine the effect of expressed protein on proliferation of HUVECs. pcDNA3.1 (+)/hCNP transfection was used for control.

RESULTSThe proliferation of HUVEC 48 h after pcDNA3.1 (+)/hCNP transfection was (0.301 +/- 0.096), which was obviously higher than that with pcDNA3.1 (+) transfection (0.164 +/- 0.012). Reconstructed pcDNA3.1 (+)/hCNP might be expressed in HUVECs effectively and its protein expression was capable of promoting HUVECs proliferation markedly.

CONCLUSIONThe successive expression of reconstructed pcDNA3.1 (+)/hCNP and the promoting activity of its expressed protein on HUVECs lay the foundation potential therapeutic value of C-type natriuretic peptide.

Cell Line ; Cell Proliferation ; Endothelial Cells ; cytology ; Humans ; Natriuretic Peptide, C-Type ; genetics ; Plasmids ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo find sensitive and specific micro-metastic markers for prostate cancer.

METHODSUsing nested reverse transcription-PCR, we examined the expression of PSA, hK2 and PSMA mRNA in peripheral blood mononuclear cells of 51 patients with prostate cancer, 33 patients with benign prostate hyperplasia (BPH) and 32 normal young people.

RESULTSThe expression rates of PSA, hK2 and PSMA mRNA were 52.9%, 43.1% and 64.7%, respectively in prostate cancer group, and 6.2%, 7.7% and 4.6%, respectively in control group (BPH patients and normal young people) with statistical significance (P < 0.01). Although the expression rate of PSA and hK2 mRNA increased with cancer progression, there was no statistical significance among patients in different stages. The expression rate of PSMA mRNA was higher than that of PSA and hK2 mRNA in each clinical stage.

CONCLUSIONPSMA mRNA expression detected by nested RT-PCR is of greater value for the diagnosis, therapy choice and prognostic evaluation of prostate cancer patients.

Aged ; Antigens, Surface ; blood ; Biomarkers, Tumor ; blood ; Glutamate Carboxypeptidase II ; blood ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; blood ; pathology ; Prostatic Neoplasms ; blood ; pathology ; Tissue Kallikreins ; blood

OBJECTIVETo investigate the influence of hypoxia induction factor-1alpha (HIF-1alpha) on glycosis of rat myocardial cell under hypoxic condition.

METHODSThe myocardial cells of the rats were routinely isolated and cultured. The cells were divided into single hypoxia (H) and HIF-1alpha inhibiting (I) groups. The cells in H group were cultured in glucose-free medium with mixed low-oxygen gas [1% O2, 94% N2 and 5% CO2 (v/v)]. While the cells in I group were cultured with low-oxygen gas after the cell model of low expression of HIF-1alpha protein constructed by RNAi technique. The cells in both groups were all observed before hypoxia (routine culture) and at the time points of 1, 3, 6, 12 and 24 hours of hypoxia. The LA (lactate acid ) content in the supernatant of the culture and the activity of the key enzymes in glycolysis such as hexokinase (HK), phosphofructokinase (PFK) and lactate dehydrogenase (LDH) of both groups of cells were determined at all the time points.

RESULTS(1) After hypoxia, the HK and PFK activities of the rat myocardial cells in H and I groups were obviously increased at the beginning and decreased thereafter when compared with that before hypoxia. While the activities of HK and PFK in H group at 1, 3 and 6 hours after hypoxia were evidently higher than those in I group (P <0.05 or 0.01), and the peak activity of them in H and I groups was 159 +/- 13 U/g vs 133 +/- 55 U/g, and 298 +/- 44 U/g vs 188 +/- 55 U/g, respectively. (2) Compared with normal control (92 +/- 12 U/g), the LDH activity of the cells in H group after hypoxia increased significantly, reaching the peak at 6 hours after hypoxia (2 568 +/- 125 U/g, P < 0. 01), and it decreased thereafter, while that in I group peaked at 3 hours after hypoxia (2125 +/- 126 U/g, P <0.01). The LA content in the culture supernatant in H group increased significantly after hypoxia with the passage of time, while that in I group increased in smaller magnitude (P <0.01).

CONCLUSIONHigh expression of HIF-1alpha in the rat myocardial cells after hypoxia could directly cause continuous enhancement of cell glycolysis, which was beneficial to the protection of myocardial cells under hypoxic condition.

Animals ; Cell Hypoxia ; Cells, Cultured ; Glycolysis ; Hexokinase ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Myocytes, Cardiac ; metabolism ; Phosphofructokinase-1 ; metabolism ; RNA Interference ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the influence of microtubule intervention drugs on the energy metabolism of myocardial cells after hypoxia.

METHODSThe primary passage of cultured myocardial cells from neonatal rats were divided into A (with hypoxia), B (with hypoxia and administration of 10 micromol/ml colchicine), C (with hypoxia and administration of 5 micromol/ml taxol), D (with hypoxia and administration of 10 micromol/ml taxol) and E (with hypoxia and administration of 15 micromol/ml taxol) groups. The creatine kinase (CK) activity and contents of ATP and ADP were assayed with colorimetry and HPLC, respectively, and the vitality of myocardial cells were determined by trypan blue method at 0.5, 1.0, 3.0, 6.0, 12.0, 24.0 post-hypoxia hours (PHH).

RESULTSThe mortality was obviously higher in B and E groups than those in A group( P < 0.05) at each time-points, but that in C and D groups were markedly lower than those in A group during 6.0 to 24.0 PHH (P < 0.01). The CK activity was significantly higher in B group than that in A group during 1.0 to 24.0 PHH, while that in E group was evidently higher, but it was lower in C and D groups than that in A group at each time-points (P < 0.05 or 0.01). The ATP contents in C group during 0.5 to 6.0 PHH were [(49.9 +/- 2.8), (40.7 +/- 2.0), (25.8 +/- 1.9), (19.1 +/- 1.2) microg/10(6) cells, respectively], which were obviously higher than those in A group [(42.9 +/- 5.8), (29.5 +/- 1.8), (18.2 +/- 0.9), (14.1 +/- 0.7) microg/10(6) cells, respectively, P < 0.05 or P < 0.01, and those in E group at each time-point were significantly lower than those in A and D groups (P < 0.01). The changes in the contents of ADP were on the contrary to the above.

CONCLUSIONMicrotubule-destabilizing drugs and high concentration microtubule-stabilizing drugs can sharply decrease ATP content in myocardiocytes under hypoxic conditions, while suitable amount of microtubule-stabilizing drugs can protect myocardiocytes by promoting its energy production.

Animals ; Cell Hypoxia ; Cells, Cultured ; Colchicine ; pharmacology ; Energy Metabolism ; drug effects ; Microtubules ; drug effects ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; Paclitaxel ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the role of p38 mitogen activated protein kinase ( p38 MAPK) in the regulation of cytosolic phospholipase A2 ( cPLA2 ) expression and degradation of membrane phospholipids in myocardium in early stage of burn rats.

METHODSWistar rats were randomized into normal group (n = 8), burn(n =40) , burn and SB203580(n = 16), burn and isotonic saline( n = 16) groups, with 8 rats at each time-points. There were 5 time-points in burn group, and 2 time-points in other groups. The rats in the latter 3 groups were inflicted with 40% TBSA full-thickness burns, and those in burn and SB203580, burn and isotonic saline groups were administered with SB203580 (p38 MAPK inhibitor) or isotonic saline, respectively. The levels of cPLA2 mRNA and membrane phospholipids in myocardium were detected with RT-PCR. In the same experiment, the effect of SB203580 on cPLA2 expression in rat myocardial cells was determined after hypoxia and burn serum treatment in vitro.

RESULTSThe level of myocardial cPLA2 mRNA in burn group at each time-point was obviously higher than those in normal group (0. 280 +/- 0. 020) , and it reached the peak value at 3 PBH. In contrast, the level of cardiac membrane phospholipids was lowered immediately after burns, and it reached the lowest level at 6 PBH [(0. 052 +/- 0. 017) mg phosphorus/mg protein]. Herein, a significant negative correlation was showed between the levels of cPLA2 mRNA and cardiac membrane phospholipids ( r = - 0. 53, P < 0. 05). Administration of SB203580, however, inhibited the increased activity of p38 MAP kinase, suppressed the upregulation of cPLA2(72% and 51% of those in burn and saline group, P <0. 01) , and markedly increased the levels of membrane phospholipids in myocardium at 6 and 12 PBH. In addition, treatment of cardiac myocytes with SB203580 also abolished the upregulation of cPLA2 mRNA elicited by hypoxia and burn serum challenge.

CONCLUSIONp38 MAP kinase play an important role in the burn-induced degradation of cardiac membrane phospholipids in rat through the upregulation of myocardial expression of cPLA2 mRNA in the myocardial cells.

Animals ; Burns ; metabolism ; Disease Models, Animal ; Myocytes, Cardiac ; metabolism ; Phospholipases A2 ; metabolism ; Phospholipids ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the effect of Shengmai injection on the management of "shock heart" after burns.

METHODSTwenty patients with severe burns were enrolled in the study and randomly divided into two groups according to the clinical research method, i.e. treatment group (n= 10, with intravenous infusion of 40 ml Shengmai injection together with 250ml 50 g/L glucose solution for 3 days, 1 time/ per day) and control group(n = 10, with intravenous infusion of 290 ml 50 g/L glucose injection liquid for 3 days, 1 time/per day). Beside the venous line used for routine fluid resuscitation for burn shock, another venous line was set up after hospitalization for the administration of the drug. Blood samples were obtained from the femoral vein in both groups at 12 post-burn hour( PBH) , and on 1, 2, 3, 4 and 5 post burn days (PBD) for the determination of serum contents of creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH) and cardiac troponin I (cTnI). The changes in hepatic and renal function, as well as coagulability were determined before drug infusion and on 1 , 2, 3, 5 and 7 PSDs.

RESULTSThe serum content of CK-MB, LDH and cTnI reached the peak at 12 PBH in both groups[ (52+/-20)U/L, (5.9+/-1.3) micromol x s(-1) L(-1), (0. 274+/-0. 231) microg/L in treatment group and [(9+/-31)U/L, (8.5+/-1l.8) micromol x s(-1) x L(-1) , (0. 584+/-0. 192) microg/L in control group]. All of them decreased with the passage of time, but in the treatment group they decreased more markedly within 2 or 3 PBD compared with those in control group ( P < 0.05).

CONCLUSIONEarly administration of Shengmai intravenously is beneficial to the protection of myocardial cells and in the management of the "shock heart" damage.

Adolescent ; Adult ; Burns ; complications ; drug therapy ; Cardiomyopathies ; prevention & control ; Creatine Kinase ; blood ; Female ; Humans ; L-Lactate Dehydrogenase ; blood ; Male ; Middle Aged ; Phytotherapy ; Prospective Studies ; Shock, Traumatic ; drug therapy ; Troponin I ; blood

OBJECTIVETo investigate the alleviation of myocardial injury of rats after early escharectomy en masse of severe burns, and to explore its molecular mechanism.

METHODSTotally 66 SD rats were randomly divided into normal control (n=6), non-escharectomy (NE, n=30) and escharectomy (E, n=30, with total escharectomy 20 minutes after burns ) groups. The rats in the NE and E groups were inflicted with 30% TBSA full-thickness scald. The content of ATP in mitochondria, troponin I (Tn I) in serum and 4.8-kb deletion of myocardial mitochondrial DNA (mtDNA) of the rats in each group were determined at 1, 3, 6, 12 and 24 post-scald hours (PSH).

RESULTS(1) The content of ATP in myocardial mitochondria was decreased in both E and NE groups, but it was obviously increased at 1 and 6 PSH (0.90 +/- 0.27 microg/mg 0.66 +/- 0.19 microg/mg) in E group when compared with those in NE group (0.74 +/- 0.18 microg/mg, 0.46 +/- 0.21 microg/mg, P < 0.05). (2) There was no obvious change in the serum content of Tn I in E group at 1 and 3 PSH, but the respective content in 1, 3 and 6 PSH was markedly lower than those in NE group (P < 0.05). (3) The 4.8 kb deletion of myocardial mtDNA was found at 1, 3, 24 PSH in NE group, while it was observed only at 1, 12 PSH in E group. The partial and whole deletion rate in E group was lower than that in NE group.

CONCLUSIONEarly escharectomy en masse can significantly alleviate the myocardial injury after burns,which might be related to its effect in lowering the deletion rate of myocardial mtDNA at early postburn stage.

Adenosine Triphosphate ; metabolism ; Animals ; Burns ; metabolism ; surgery ; DNA, Mitochondrial ; genetics ; Disease Models, Animal ; Female ; Male ; Mitochondria, Heart ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Sequence Deletion ; Troponin I ; blood

OBJECTIVETo investigate the protective effect of glycine (Gly) on hypoxic rat myocardial cells and its mechanism.

METHODSSdfetal rat myocardial cells were isolated and cultured in vitro. The released amounts of creatine kinase (CK) and lactate dehydrogenase (LDH) from the myocardial cells in the culture supernatant at 6 hour after hypoxia and after glycine treatment were determined with ultraviolet spectrophotometer. The expression of the alpha1 subunits of glycine receptor (GlyRalpha1) in the myocardial cells was detected by immunofluorescent histochemistry. The changes in the intracellular calcium content and the membrane potential of the myocardial cells were determined by laser confocal microscopy.

RESULTSThe release of CK and LDH in the culture supernatant increased significantly at 6 h after hypoxia [(393.8 +/- 5.3), (1564 +/- 41) U/L] compared with those before hypoxia, while their levels were obviously decreased after glycine treatment [(56.3 +/- 2.7), (716 +/- 18) U/L, (P <0.01)] compared with those before glycine treatment. There was positive expression of GlyRalpha1 in myocardial cells before and after hypoxia. The average fluorescent intensity of intracellular calcium at 6 hours after hypoxia (139 +/- 29) was significantly higher than that before hypoxia (27 +/- 8, P < 0.01), while it was obviously lower (51 +/- 11) after glycine treatment compared with that at 6 hours after hypoxia,but it was evidently higher than that before hypoxia (P <0.01). The membrane potential 6 hours after hypoxia (62 +/- 9) was obviously lower than that before hypoxia (177 +/- 20, P < 0.01), but it was obviously higher after glycine treatment (123 +/- 16) than that at 6 hours after hypoxia (P < 0.01).

CONCLUSIONGlycine might be beneficial in the protection of myocardial cells against hypoxia. The underlying mechanism may involve attenuation of membrane potential depolarization after hypoxia by conjugation of glycine with its receptor, depleting in turn voltage-dependent calcium channel on the cellular membrane, preventing calcium overload due to influx of calcium ions after hypoxia.

Animals ; Calcium ; metabolism ; Cell Hypoxia ; drug effects ; Cells, Cultured ; Creatine Kinase ; metabolism ; Glycine ; pharmacology ; L-Lactate Dehydrogenase ; metabolism ; Membrane Potentials ; drug effects ; Myocytes, Cardiac ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of the Chinese herbal medicine of Longbixiao (LBX) Capsule on the expressions of TGF-beta1 and Smoothelin in human prostatic stromal cells cultured in vitro.

METHODSBlood serum medicated with LBX was incubated with the stromal cells isolated from men with benign prostatic hyperplasia (BPH) and cultured in vitro. The mRNA expression levels of TGF-beta1 and Smoothelin were detected by real-time RT-PCR and other relevant techniques.

RESULTSIn the high and low concentration groups, the gene relative expressions of TGF-beta1 were (0.158 +/- 0.020) and (0.169 +/- 0.020) , while those of Smoothelin were (0.035 +/- 0.007) and (0.036 +/- 0.007) respectively, both significantly decreased in comparison with the control group(P < 0.01).

CONCLUSIONLBX reduces the mRNA expressions of TGF-beta1 and Smoothelin in human prostatic stromal cells and can be used in the treatment of BPH.

Animals ; Capsules ; Cells, Cultured ; Cytoskeletal Proteins ; genetics ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gene Expression ; drug effects ; Humans ; Male ; Muscle Proteins ; genetics ; Prostatic Hyperplasia ; pathology ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Serum ; chemistry ; Stromal Cells ; drug effects ; metabolism ; pathology ; Transforming Growth Factor beta1 ; genetics

OBJECTIVETo explore the underlying mechanism of mesenchymal stem cells (MSCs) transfer induced cardiac function improvement in failing hearts.

METHODSCongestive heart failure (CHF) was induced in rats by cauterization of the heart wall. MSCs were cultured from autologous bone marrow and injected into the border zone and the remote myocardium 5 days after cauterization.

RESULTSTen weeks later, cardiomyocyte nucleus mitotic index, capillary density and expression of insulin-like growth factor 1 (IGF-1), hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) were significantly increased in the border zone and significantly reduced in the remote myocardium in CHF rats (all P<0.05 vs. sham). Besides cardiac function improvement and left ventricular remodeling attenuation evidenced by hemodynamic and echocardiographic examinations, expressions of IGF-1, HGF and VEGF in the remote myocardium and in the border zone were also significantly upregulated (P<0.05 or P<0.01 vs. CHF), and cardiomyocyte nucleus mitotic index as well as capillary density were significantly increased in CHF rats with MSCs (P<0.05 or P<0.01 vs. CHF). Moreover, collagen area was significantly reduced and myocardial area was significantly increased in the border zone in these rats too.

CONCLUSIONAutologous MSC implantation upregulated expressions of growth factors enhanced cardioangiogenesis which might be the underlying mechanisms for improved cardiac function and attenuated left ventricular remodeling induced by MSCs transplantation in failing rat myocardium.

Animals ; Disease Models, Animal ; Heart Failure ; metabolism ; therapy ; Hepatocyte Growth Factor ; metabolism ; Insulin-Like Growth Factor I ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; metabolism ; Ventricular Remodeling