Objective To investigate whether high glucose can induce endothelial-mesenchymal transition (EndMT) in glomerular endothelial cells and the role of TGF-β in the process.Methods Rat glomerular endothelial cells were divided into five groups:normal glucose (NG,5.5mmol/L),high glucose (HG,15,30 mmol/L),TGF-β inhibition (HG+ LY36,30 mmol/L glucose + 10 μmol/L LY364947),hyperosmotic control (M,5.5 mmol/L glucose+25.5 mmol/L mannitol) and solvent control (D,5.5 mmol/L glucose + 1 ml/L DMSO).Western blotting was performed to detect relative protein quantities of endothelial marker claudin 5 and mesenchymal marker α-smooth muscle actin (α-SMA).TGF-β1 and TGF-β2 mRNA levels were measured by real-time PCR.Vascular endothelial marker VE-cadherin and mesenchymal marker α-SMA were detected by immunofluorescent stain and observed by confocal microscopy.Results Compared with NG,the expression of claudin5 protein in HG (15 or 30 mmol/L) was up-regulated while expression of α-SMA protein was down-regulated (P ＜0.05).Both TGF-β1 and TGF-β2 mRNA levels increased as well (P ＜ 0.05).However,when compared with HG,the claudin 5 levels increased while α-SMA decreased in TGF-β inhibition group.No significant changes were observed in hyperosmotic or solvent control group.Confocal microscopy showed the transformation of cells from a cobblestone-liked shape to a spindle one,and a decreasing expression of VE-cadherin while an increasing α-SMA in HG group (P ＜ 0.05),whereas TGF-β inhibition partly attenuated those changes in both morphological and protein levels.Conclusions High glucose treatment of glomerular endothelial cells results in an increase in the level of TGF-β1 and TGF-β2 mRNA and leads to endothelial-mesenchymal transiton.Inhibition of TGF-β partly prevents this process,indicating that TGF-β plays a crucial role in high-glucose-induced glomerular endothelial-mesenchymal transiton.

ObjectiveTo explore the value of virtual touch tissue quantification (VTQ) in the evaluation of clinical pathological typing of advanced gastric cancer.MethodsFifty six patients who had been diagnosed as gastric cancer were examined using acoustic radiation force impulse.According to clinical pathological typing,all cases were divided into highly-moderately differentiated adenocarcinoma (14 cases) and non-highly-moderately differentiated adenocarcinoma (42 cases).A comparison with clinical pathologic results was made after surgery.The correlation of VTQ results and clinical pathological typing of gastric cancer was analyzed.ResultsThe VTQ value of highly-moderately differentiated adenocarcinoma was lower than that of non-highly-moderately differentiated adenocarcinoma [(1.49 ± 0.44) m/s vs (2.12 ± 0.45) m/s],with statistical significance( t =-4.53,P ＜0.05).According to the maximum area under the ROC curve,the cutoff value of VTQ was 1.795 m/s,the sensitivity and specificity were 78％ and 86％,respectively,the Youden's index was 0.64,and with high reproducibility(Kappa =0.81).ConclusionsVTQ could initial estimate the clinical pathological typing of advanced gastric cancer before operation.

ObjectiveTo explore the value of virtual touch tissue quantification (VTQ) in evaluating Lauren classification of advanced gastric carcinoma.MethodsForty-one patients with gastric cancer proved by endoscopic biopsy performed preoperative VTQ examination,and the findings were compared with postoperative pathologic results via hematoxylin -eosin and Alcian Blue-Periodic Acid Schiff (AB-PAS) staining.ResultsIn 41 patients,26 cases were diagnosed as diffuse type and 15 cases as intestinal type by pathology after operation.The shear wave velocity(SWV) of diffuse type was higher than that of intestinal type [(1.72± 0.83)m/s vs (1.05± 0.66)m/s,t =2.819,P=0.008] measured by virtual touch tissue quantification (VTQ).According to the area under the ROC curve,the cut- off value of SWV in gastric cancer tissues for assessing the diffuse type was 1.045 m/s with a sensitivity of 80.8％ and specificity of 73.3 ％ respectively.ConclusionsVTQ could be considered as a promising method to distinguish the Lauren classification in patients with advanced gastric carcinoma.

Objective To explore the changes of the plasma motilin(MTL)and serum gastrin(GAS)levels and their relationship with course of disease,the severity,the complication of digestive system in neonates with intracranial haemorrhage(ICH).Methods The objects were 26 cases of term newborns with ICH,the healthy control group contained 30 cases of healthy term newborns.Plasma motilin and serum gastrin concentration were measured by radioimmmunoassay in 26 cases of newborns with ICH in the 2 d,3-5 d,7-10 d,and 12-15 d after birth,and compared with the healthy control group.Results Compared with healthy control group,the blood GAS and MTL levels of neonatal ICH group increased dramatically(Pa

Objective To explore the relationship between dynamic changes in ultra-micro-structural of pulmonary arteries and endogenous hydrogen sulfide in rats with left-right shunt.Methods Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of pulmonary artery structural remodeling. After 1 day, 3 days, 1 week, 4 weeks and 8 weeks of experiment, the ultra-micro-morphologic changes of pulmonary arteries of rats were observed under electronic microscope and H_2S concentration in serum was evaluated by modified sulfide electrode method.Results The changes of ultra-micro-structure of pulmonary arteries were progressively exacerbated, endothelial cells became swollen and large in size on 3 days, smooth muscular cells increased in size as well as the change of endothelial cells in 1 week, and they changed from contractile phenotype to synthetic phenotype in 4 weeks.Conclusions Shunt exhibited changes of ultra-micro-structure of pulmonary arteries are accompanied by the changes of endogenous H_2S. It is suggested that endogenous H_2S might play a protective role in changes of ultra-micro-structure of pulmonary artery.

OBJECTIVETo evaluate the efficacy and safety of the parenteral administration of various quantities of amino acid in preterm infants.

METHODSPreterm infants (birth weight 1000-2000 g) recruited into the study were randomized into three groups. High amino acid group (HP): 2.4 g/(kg.d) of amino acid IV within 24 hours after birth increasing by increments of 1.2 g/(kg.d) to a maximum of 3.6 g/(kg. d); medium amino acid group (MP): 1.0 g/(kg.d) of amino acid IV 24 hours after birth, increasing by increments of 0.5 g/(kg.d) until a maximum of 3.0 g/(kg.d); and low amino acid group (LP): 0.5 g/(kg.d) of amino acid on D3, increasing by increments of 0.5 g/(kg.d) until a maximum of 3.0 g/(kg.d) as the final dose.

RESULTSTotally 96 preterm infants were recruited: HP 34, MP 32 and LP 30. There were no significant differences in demographic or clinical characteristics among the 3 groups. HP group showed lower postnatal weight loss (43.4 g, 95% CI 74.3, 12.6) and weight loss% (2.84%, 95% CI 4.79%, 0.71%) than LP group. HP group showed shorter length of stay in NICU (5.25 d), days to reach 2000 g (7.03 d) and days to tolerate 100 kcal/(kg.d) enteral nutrition (4.52 d) than LP group. Cost of hospitalization was significantly lower in HP group than in LP group (-6275 RMB, 1 US$=8 RMB) and MP group (-5715 RMB). Mean serum RBP (D4), threonine and tyrosine levels were significantly higher in HP group than in LP group. Serum insulin levels were similar; mean serum glucose level was lower in HP group than in LP group. HP infants had lower incidence of sepsis than LP infants (21.9% vs 40.0%). There were no significant differences in the levels of blood ammonia, acid-base balance (as determined by pH and NaHCO3-), BUN, Cr, AST, and ALT.

CONCLUSIONSIntensive and early administration of intravenous amino acid [2.4 g/(kg.d)] improves preterm infants' growth and the tolerance of enteral feeding. It also reduces the cost of hospitalization, and the incidence of sepsis.

Amino Acids ; administration & dosage ; adverse effects ; Humans ; Infant, Newborn ; Infant, Premature ; Parenteral Nutrition

Objective:To find the best synthesis method of 6-benzyl-1-[ ( benzyloxy ) methyl ]-3-hydro-xy-5-(hydroxymethyl)pyrimidine-2,4(1H,3H)-dione e for observing the change of its biological activity after N-3 hydroxylation .Methods:After trying some N-hydroxylation methods , the target compound was successfully synthesized via one-pot oxidizing process by sodium hydride ( NaH) and 3-chloroperbenzoic acid( m-CPBA);the anti-HIV reverse transcriptase ( RT) activity and integrase ( IN) activity of the tar-get compound was assayed via enzyme-linked immunesorbent assay ( ELISA) and phosphorylation of DNA package method .Results:The target compound could be obtained through the improved m-CPBA oxida-tive method by only one step , and the yield of the reaction could reach 60%-70%.And the structure of this compound was identified by 1 H NMR, 13 C NMR and MS;The activity result showed it added the an-ti-HIV IN activity after N-3 hydroxylation as well as retained the anti-HIV RT activity.Conclusion:The improved m-CPBA oxidative method is a convenient and efficient way to prepare the compound 6-benzyl-1-[(benzyloxy)methyl]-3-hydroxy-5-(hydroxymethyl)pyrimidine-2,4(1H,3H)-dione e which has both anti-HIV RT and IN activity .

OBJECTIVEHypoxic pulmonary hypertension is an important pathophysiologic process of various cardiovascular diseases. Sulfur dioxide (SO2) was considered as a kind of toxic gas previously, but recent studies suggested that SO2 could act as a key bioactive molecule in the pathogenesis of cardiovascular diseases. Therefore, this study was designed to examine the effect of sulfur dioxide on pulmonary vascular structure of hypoxic pulmonary hypertensive rats treated with SO2 donor substances.

METHODSThe rats were randomly divided into 3 groups: control group(n = 8), hypoxic group(n = 8) and hypoxic + SO2 group (n = 10, treated with SO2 donor Na2SO3/NaHSO3). The rats of hypoxic group and hypoxic + SO2 group were under a hypoxic condition for 21 days, while the rats of control group were exposed to room air. The mean pulmonary artery pressure was tested by means of right cardiac catheterization and the content of SO2 in plasma was investigated by high performance liquid chromatography (HPLC). The change in relative medial thickness (RMT) of pulmonary arteries was examined under optical microscope. The ultra-structural changes were observed under a transmission electron microscope. The data were analyzed through one-way analysis of variance (ANOVA) by SPSS 13.0 software.

RESULTSCompared with control group [(2.25 +/- 0.50) kPa], the mean pulmonary artery pressure of hypoxic group [(5.12 +/- 0.51) kPa] raised significantly (t = 5.091, P < 0.01) and RMT of hypoxic group (9.66 +/- 1.27) compared with control group (6.83 +/- 1.57) significantly raised (t = 3.392, P < 0.01). Ultrastructural observation showed the proliferation and degeneration of endothelial cells in small pulmonary arteries in rats with pulmonary hypertension. The internal elastic lamina was irregular. The proliferation of medial smooth muscle cells of arteries was shown at the level of respiratory bronchioles. The collagens also increased. Meanwhile, compared with control group [(33.36 +/- 5.62) micromol/L], the content of SO2 in plasma of hypoxic group [(27.01 +/- 4.17) micromol/L] declined (t = 2.067, P < 0.05). Whereas compared with that of hypoxic group [(5.12 +/- 0.51) kPa], the mean pulmonary artery pressure of hypoxic + SO2 group [(3.94 +/- 0.33) kPa] declined (t = 2.712, P < 0.01) and RMT of hypoxic + SO2 group (6.97 +/- 1.83) decreased compared with hypoxic group (9.66 +/- 1.27) (t = 3.009, P < 0.01). Compared with those of hypoxic group, the pulmonary artery ultrastructural changes in hypoxic group ameliorated obviously after using exogenous sulfur dioxide donor. The endothelial cells became flat and the smooth muscle cells of arteries slightly enlarged and arranged regularly. At the same time, compared with hypoxic group [(27.01 +/- 4.17) micromol/L], the content of SO2 in plasma of hypoxic + SO2 group [(29.89 +/- 4.52) micromol/L] raised (t = 1.263, P > 0.05).

CONCLUSIONSulfur dioxide plays an important role in the regulation of small pulmonary artery structural changes in hypoxic pulmonary hypertensive rats. The hypoxic pulmonary hypertensive damages can be ameliorated significantly after using exogenous SO2 donor.

Animals ; Hypertension, Pulmonary ; blood ; pathology ; physiopathology ; Hypoxia ; blood ; pathology ; physiopathology ; Male ; Pulmonary Artery ; drug effects ; pathology ; Rats ; Rats, Wistar ; Sulfur Dioxide ; adverse effects ; blood

OBJECTIVEPulmonary vascular structural remodeling induced by high pulmonary blood flow is an important pathologic basis of pulmonary hypertension with congenital heart disease of left-to-right shunt. However, the mechanism is still not clear. The present study aimed to examine the alteration of endogenous nitric oxide (NO) pathway in high pulmonary blood flow-induced pulmonary vascular structural remodeling, so as to explore the role of NO pathway in pulmonary hypertension induced by high pulmonary blood flow.

METHODSSixteen male SD rats were randomly divided into control group (n = 8) and shunting group (n = 8). Aortocaval shunting was produced for 11 weeks in shunt rats. Pulmonary artery mean pressure (mPAP) of each rat was evaluated using right cardiac catheterization. The ratio of right ventricular mass to left ventricular plus septal mass [RV/(LV + S)] was detected. Pulmonary vascular micro-and ultra-structure was examined by using a light microscope and a transmitted electronic microscope. Meanwhile, the concentration of plasma NO was measured by spectrophotometry. The expressions of endothelial NO synthase (eNOS) mRNA and protein by pulmonary arteries were detected by in situ hybridization and immunohistochemistry, respectively.

RESULTSAfter 11-week aortocaval shunting, mPAP was significantly increased [(22.5 +/- 2.6) mmHg vs. (15.8 +/- 2.8) mmHg, 1 mmHg = 0.133 kPa, t = 4.97, P < 0.01], and RV/(LV + S) was also markedly increased (0.267 +/- 0.022 vs. 0.221 +/- 0.016, t = 4.85, P < 0.01). The percentage of muscularized arteries was obviously increased in shunt rats compared with controls [(23.2 +/- 2.4)% vs. (13.5 +/- 2.1)%, t = 7.82, P < 0.01], and relative medial thickness of pulmonary arteries was obviously increased in shunt rats [median pulmonary artery: (7.76 +/- 0.56)% vs. (4.82 +/- 1.03)%, t = 6.23, P < 0.01; small pulmonary artery: (11.94 +/- 0.66)% vs. (6.91 +/- 0.53)%, t = 14.96, P < 0.01]. Ultrastructural changes, such as hyperplasia and degeneration of endothelial cells, irregularity of internal elastic laminar and hypertrophy and the increased number of synthetic phenotype of smooth muscle cells, were found in intrapulmonary arteries of shunt rats. Meanwhile, plasma NO concentration was increased [(30.2 +/- 7.9) micromol/L vs (19.7 +/- 5.7) micromol/L, t = 3.05, P < 0.01) and eNOS mRNA and protein expressions by pulmonary arteries were significantly augmented in rats of shunting group.

CONCLUSIONThe upregulation of eNOS/NO might be an adaptive response of pulmonary circulation to an increased blood flow in the development of pulmonary hypertension and pulmonary vascular structural remodeling.

Animals ; Blood Flow Velocity ; Hypertension, Pulmonary ; physiopathology ; Immunohistochemistry ; In Situ Hybridization ; Male ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; genetics ; Nitric Oxide Synthase Type III ; Pulmonary Artery ; physiopathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

UNLABELLEDTo explore the changes of time-dependent pulmonary artery structural remodeling in pulmonary hypertension induced by high pulmonary flow in rats.

METHODSEighty male SD rats were randomly divided into control group (n =40) and shunt group (n = 40). Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. In the control group, rats experienced the same experimental processes except the shunting procedure. After 1 day, 3 days, 1 week, 4 weeks, and 8 weeks of experiment, systolic pulmonary artery pressure (SPAP) and mean pulmonary artery pressure (MPAP) of each rat were evaluated by using a right cardiac catheterization procedure. Heart tissues were separated as right ventricle (RV) and left ventricle plus septum (LV+SP), and the ratio of RV to LV+SP [RV/ (LV+ SP)] was calculated. The morphologic changes including micro- and ultra-structural changes of pulmonary arteries of rats were observed under optical microscope and electro-microscope, respectively. The percentages of muscularized artery (MA), partial muscularized artery (PMA) and non-muscularized artery (NMA) in small pulmonary arteries and median pulmonary arteries were calculated. The changes of relative medial thickness (RMT) and relative medial area (RMA) of pulmonary arteries were examined.

RESULTSCompared with control group, SPAP and MPAP did not change on day 1, day 3, and week 4. However, in week 1 and week 8 of experiment, SPAP and MPAP increased significantly (P < 0.01). Meanwhile, in week 8 of experiment, RV/ (LV+SP) increased significantly (P < 0.05). In contrast to control group, the percentages of MA, PMA, and NMA did not change in day 1, day 3 and week 1. But in week 4 and week 8, the percentages of MA and PMA increased significantly (P < 0.01) but that of NMA decreased significantly (P < 0.01). RMT and RMA did not change in day 1, day 3, week 1 and even week 4 in shunt group as compared with those of control group, but they increased significantly in week 8 (P < 0.05). The changes of ultra-structure of pulmonary arteries included that endothelial cells became swollen and large in size on day 3, smooth muscular cells increased in size besides the change of endothelial cells in week 1, and they changed from contractile phenotype to synthetic phenotype in week 4. Collagen deposited in pulmonary arteries markedly in week 8.

CONCLUSIONPulmonary artery structural remodeling develops in a time-dependent manner. Endothelial cells of pulmonary arteries become swollen firstly, followed by the proliferation of smooth muscular cells and finally by remodeling of extra cellular matrix.

Animals ; Blood Pressure ; Hypertension, Pulmonary ; pathology ; physiopathology ; Male ; Pulmonary Artery ; pathology ; ultrastructure ; Pulmonary Circulation ; Random Allocation ; Rats ; Rats, Sprague-Dawley