Objective To explore the role of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in hyperoxia-induced lung injury in preterm rats. Methods At the 2 nd postnatal day Sprague-Dawley preterm rats were randomly assigned to air group and hyperoxia group (exposed to about 85% of O 2). At 3,7,14 and 21 days after exposure, six rats of each group were used to assess lung histologic changes and expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 in lungs by immunohistochemistry. At 3,7 and 14 days after exposure, gelatinase activity in bronchoalveolar lavage fluid (BALF) of another six rats in each group by gelatin zymography was examed. Results Except 3 d after exposure, hyperoxia group showed lung injury characterized by subacute alveolitis and inhibition of lung development. Expression of MMP-2, MMP-9, TIMP-1 and TIMP-2 in hyperoxia group was stronger than that in air group at every time (P

The effect of extraction and nonextraction treatment on the soft tissue profile of the subjects borderline Angle classⅡ division 1 malocclusion is concerned mainly by orthodontists.This article reviewes the effect of extraction and nonextraction treatment on the soft tissue profile of the subjects with Angle classⅡ division 1 borderline patients and the important factors(such as differences between the types of tooth extraction and growth).The aim is to guide clinic diagnosis and treatment for borderline Angle classⅡ division 1 malocclusion.

Objective To establish a method of isolation, purification and identification of type Ⅱ alveolar epithelial cells (AEC- K ) from neonate piglet lungs of 1 ～ 3 days old and to investigate effects of proinflammatory cytokines on expression of growth factors (GFs). The yield, viability and purity of AEC- Ⅱ obtained using different enzyme digestion and purifying methods were compared. Methods After the first 24-hour culture of AEC- Ⅱ ,the media containing interleukin (IL)-1β,IL-6 and IGF-Ⅰ at different concentrations were used to culture AEC-Ⅱ for another 48 hours. And then the cells were counted and the expressions of insulin-like growth factor (IGF-Ⅰ ), platelet-derived growth factor ( PDGF), surfactant proteins (SP) -A and SP-B mRNA were determined by real time PCR. Results A significantly higher yield of AEC-Ⅱ was achieved by digesting the lung with 30 unit/ml elastase and 0.1 % trypsin at 37 t for 20 min, the yield was (5.33 ±0.54) × 106 after adjusted by the weight of lung and heart (P <0.01). The number of purified AEC-II obtained by immune adherence method was (38.0 ±28.0) × 106 perpiglet which was higher than by the method of percoll. The optimal phenotype maintenance time of AEC- Ⅱ was the first 24～96 hours in the primary culture. With increasing concentrations of IL-1 β and IL-6, there were decreased proliferation and expression of SP-A and IGF-Ⅰ mRNA in the cultured AEC- Ⅱ ,but SP-B mRNA expression was not affected. Both AEC-Ⅱ proliferation and expression of SP-A, SP-B mRNA decreased significantly after cultured with anti-IGF-Ⅰ. Conclusion In a new model of cultured AEC-Ⅱ from neonate piglets, IL-1β and IL-6 inhibited AEC- Ⅱ proliferation and SP-A mRNA expression through IGF-Ⅰ -dependent mechanisms.

Purpose To study the hemodynamic changes of kidney interlobar artery in children with type 1 diabetes, and to explore the clinical value of the color Doppler ultrasound in diabetic nephropathy during the incubation period. Materials and Methods The study was performed in 52 children with type 1 diabetes and in 51 age-matched healthy children as control group. The resistance index (RI) of interlobar artery was measured with color Doppler ultrasonography, and the clinical data and laboratory indicators were analyzed. Results The left kidney RI, right kidney RI and average RI were all significantly higher in children with diabetes than in age-matched healthy group (P<0.01). The mean RI were correlated positively with (HbA1c) (r=0.96, P<0.01) and diabetes duration (r=0.31, P<0.01), respectively. Fasting blood glucose, glycated hemoglobin, and glomerular filtration rate were significantly higher in children with diabetes than in age-matched healthy group (P<0.01). Conclusion The RI of interlobar artery increases in children with type 1 diabetes. Early changes of renal hemodynamics in children with type 1 diabetes are detectable with Doppler sonography, and have certain value in early diagnosis of diabetic nephropathy.

Objective To study the effect of transplanted endothelial progenitor cell (EPC) on hyperoxia-induced lung injury in neonatal rats.Methods Rat bone marrow mononuclear cells were cultured in endothelial cell growth medium to obtain EPCs,which were identified by morphology,phagocytosis and CD34+ analyses.Sixty neonatal Sprague-Dawley rats were allowed to acclimate in room air for 24 h after birth,and were then divided into four groups (15 per group),including the air group,the hyperoxia group,the EPCs transplantation group and the N ω-nitro-L-arginine methyl ester (L-NAME) intervention group.Neoborn rats in the Air and Hyperoxia groups were fed in the room air or hyperoxia (85％ oxygen) for 28 days.For rats in transplantation group were exposed continuously to hyperoxia for 28 days,and got an EPC (1 × 105 cells) injection on the 21st day.Rats in Intervention group were exposed continuously to hyperoxia for 28 days,got an EPC (1 × 105 cells) injection on the 21st day,and a daily injection of L-NAME from day 21 to day 28,with a daily dose of 20 mg/kg.Levels of circulating CD34+ cells and serum VEGF expression were detected.Specimens from lung tissues were analyzed by immunohistochemistry or immunofluorescence.The expression of vascular endothelial growth factor (VEGF),VEGF receptor 2 (VEGFR2) and eNOS were detected by realtime polymerase chain reaction and Western-blotting.NO production were detected by nitrate reductase assay.One way ANOVA and Bonferroni test were used for statistical analysis.Results (1) The cultured cells had a typical cobblestone appearance; double positive cell binding of fluorescein Ulex Europaeus agglutinin-1 and uptake of Dil-labeled acetylated low density lipoprotein accounted for approximately 85％ of the total number of cells.CD34+ cells accounted for 68.2％-72.4％ of total cultured cells.(2) Circulating CD34+ cells in the air group,hyperoxia group,EPC transplantation group and L NAME intervention group were (1.91 ± 0.34)％,(1.06 ± 0.10)％,(1.47 ± 0.06)％ and (0.77 ± 0.11)％ (F=32.710,P=0.000).The number of circulating CD34+ cells in the hyperoxia group was lower than the air group,in the EPC transplantation group the number of these cells was higher than the hyperoxia group,and in the L-NAME intervention group the number of these cells was lower than that in the EPC transplantation group,and the differences between these two groups were statistically significant (P ＜ 0.05,respectively).Serum VEGF in the four groups was (7.90±2.72),(6.38±0.72),(14.00± 1.66) and (11.70± 1.91) pg/ml,respectively.The difference between the four groups was statistically significant (F=22.809,P=0.000),and serum VEGF in the EPC transplantation group was higher than that in the hyperoxia group (P ＜ 0.05).(3) Transplanted EPCs could engraft in pulmonary vascular endothelium and alveolar interstitium,and L-NAME intervention significantly reduced the engraftment of EPCs in the lungs (10.7±0.47 / field vs 16.95±0.5 /field,t=17.820,P=0.000).(4) There were significant differences in the radial alveolar count (RAC) and number of microvessels between the four groups (F=859.580 or 211.150,P=0.000,respectively).RAC and the number of microvessels in the hyperoxia group were less than those in the air group (7.98±0.23 vs 13.12±0.20,3.98±0.42 vs 9.50±0.22,P ＜ 0.05,respectively).The number of microvessels in the EPC transplantation group was 5.40±0.41,being higher than that in the hyperoxia group (P＜0.05).(5) VEGF mRNA in lungs in the hyperoxia group was lower than that in the air group (0.23 ± 0.16 vs 1.05 ± 0.33,P ＜ 0.05); in the EPC transplantation group,VEGF mRNA was higher than that in the hyperoxia group (0.69 ± 0.09 vs 0.23 ± 0.16,P ＜ 0.05); and in the L-NAME intervention group,VEGF mRNA was lower than that in the EPC transplantation group (0.31 ±0.08 vs 0.69±0.09,P ＜ 0.05).VEGF protein in the lungs in the hyperoxia group was lower than the air group (0.52±0.01 vs 0.82±0.01,P ＜ 0.05),and was higher in the EPC transplantation group than the hyperoxia group (0.58±0.05 vs 0.52±2501,P ＜ 0.05).VEGFR2 mRNA in the hyperoxia group was lower than the air group (0.35±0.13 vs 1.07±0.45,P ＜ 0.05).eNOS mRNA in the hyperoxia group was lower than the air group (0.46±0.10 vs 1.05±0.36,P ＜ 0.05).eNOS protein in the hyperoxia group was lower than the air group (0.32±0.01 vs 0.51 ±0.03,P ＜ 0.05),and was higher in the EPC transplantation group than the hyperoxia group (0.86±0.02 vs 0.32±0.01,P ＜ 0.05).Conclusion Transplanted EPC can engraft in the lung tissue,improving alveolar and pulmonary vascular development,which may be associated with upregulation of the expression of eNOS and VEGF in lung.

AIM: To investigate the therapeutic effect of endothelial progenitor cell-conditioned medium (EPC-CM) on the lung structure of neonatal rat exposed to hyperoxia, and to explore the mechanisms.METHODS: Bone marrow-derived endothelial progenitor cells (EPCs) were collected from new born Sprague-Dawley (SD) rats and the EPCs were identified.The conditioned medium from the passage 3 EPCs was collected.Newborn SD rats (n=40) were randomly divided into 4 groups.The rats in room air group were exposed to the room air (21% O2) for 21 d.The rats in hyperoxia group were exposed to hyperoxia (85% O2) for 21 d.The rats in endothelial cell basal medium (EBM) group were exposed to hyperoxia for 21 d, and received 100 μL EBM on postnatal day 14 (P14) in a single intratracheal (IT) injection.The rats in EPC-CM group were exposed to hyperoxia for 21 d, and received 100 μL EPC-CM on P14 in a singlie IT injection.The rats were sacrified on the 21st day.The left lungs were excised, placed in 4% paraformaldehyde, serially dehydrated in ethanol and embedded by paraffin.Serial sectioning of the paraffin-embedded left lung tissues was prepared for 5 μm thickness, and stained with hematoxylin and eosin.The pulmonary radical alveolar count (RAC) and alveolar mean linear intercept (MLI) were then calculated.The microvascular density was determined by FVIII immunostaining.The mRNA expression of KGF, VEGF, SP-A and SP-C in the right lung tissues was detected by real-time fluorescence quantitative PCR.RESULTS: The cultured cells had typical EPC morphological characteristics, and had the abilities to bind to FITC-UEA-1 and uptake DiI-ac-LDL.The body weight of the rats on day 21, RAC, MLI and microvascular density were significantly lower in hyperoxia group and EBM group than those in room air group (P<0.05).The EPC-CM group had significantly higher RAC and microvascular density than those in hyperoxia group and EBM group (P<0.05), but the body weight and MLI had no significant difference.The mRNA expression levels of KGF, VEGF, SP-A and SP-C in hyperoxia group and EBM group were significantly lower than those in room air group (P<0.05).The mRNA expression levels of KGF, VEGF, SP-A and SP-C in EPC-CM group were significantly higher than those in hyperoxia group and EBM group (P<0.05).CONCLUSION: EPC-CM promotes the lung alveolarization and microvascular formation in neonatal rats exposed to hyperoxia.These benefits may be correlated with the increased KGF and VEGF mRNA expression.

Objective To investigate the relationships between septic shock and vascular endothelial cells (VECs)injury by measuring the plasma CEC,vWF,and NO in septic shock patients and to explore its potential clinical values. Methods 72 patients were divided into group A including 34 infection patients but without septic shock,and group B including 38 patients with septic shock. Circulating endothelial cell(CEC),von willebrand factor(vWF),and nitric oxide(NO)in plasma were measured in all patients at the time points of accessing ICU (T0)and 24 hours after staying in ICU(T1). And the blood pressures of studied patients were also recorded at the same time points. Results The levels of CEC and vWF at each observing time point were higher in group B than those in group A(P < 0.01),while the levels of NO were lower in group B than those in group A(P < 0.01). In group B,the levels of CEC and vWF at T1 were higher than those at T0(P<0.01),while the level of NO was low-er at T1 than that at T0(P<0.01). With contrary to those of group A,circulation conditions of group B were obvi-ously unstable and high doses of vasoactive agents were needed during the therapy period. MAP in group B at each time point was obviously lower than that in group A(P<0.05). There were 7 dead cases(18%)in group B but no death in group A during the whole therapy period. Conclusion Obvious injury of vascular endothelial cells exists in septic shock patients and the injury degree is related to the severity of the disease.

Objective To investigate the role of tet methylcytosine dioxygenase 2 (TET2) in the regulation of transforming growth factor-β1 (TGF-β1) expression in human glomerular mesangial cells induced by high glucose.Methods Cultured human glomerular mesangial cells were divided into normal control group (5.5 mmol/L glucose) and high glucose group (30.0 mmol/L glucose) which was cultured for 12 h to 72 h.The gene expression of TET2 in mesangial cells were inhibited by small molecule chemical called SC1,and which were divided into high glucose group (30.0 mmol/L glucose+ DMEM),DMSO group (30.0 mmol/L glucose+0.1％DMSO) and SC1 group (30.0 mmol/L glucose+3 μmol/L SC1).The mRNA and protein expression of TGF-β1,TET1 to 3 and α-smooth muscle actin (α-SMA) was detected by quantitative real-time PCR and Western blotting.Methylation of CpG islands in the regulation region of TGF-β1 was detected by bisulfite sequencing PCR (BSP).The activity of mesangial cell proliferation was assessed by colorimetry of thiazolyl blue (MTT).Results Compared with normal control group,the mRNA and protein expression of TET2 in mesangial cells induced by high glucose was increased significantly in a time-dependent manner (all P ＜ 0.05),but the expression of TET1 and TET3 was not affected.Meanwhile methylation rate of 4 CG sites from 24 h to 72 h were decreased in the first exon of TGF-β1 (P ＜ 0.01),but not in the promoter.Compared with high glucose group,when the expression of TET2 was inhibited by SC1,the methylation rate of TGF-β1 was recovered evidently (P ＜ 0.05),the mRNA and protein expression of TGF-β1 and α-SMA was suppressed,and the proliferation of mesangial cells was decreased (all P ＜ 0.05).Conclusions Demethylation of the CpG island mediated by TET2 may play an important role in the expression of TGF-β1 and mesangial cell phenotype transformation induced by high glucose.

ObjectiveTo establish a method for measuring exhaled nitric oxide (eNO)concentrations in neonates with and without hypoxemic respiratory failure ( HRF), and to investigate the relationship between eNO and respiratory parameters in neonates with HRF. Methods Twenty-two newborn infants with HRF and 26 control neonates were included within the first 24 hours of postnatal life.Their eNO levels were detected with a rapid-response chemiluminescence analyzer daily during the first week of their postnatal life, and lung mechanics and gas exchange efficiency were monitored at the same time, such as pulse oxygen saturation ( SpO2 ), inspired fraction of oxygen ( FiO2 ) and other parameters.Wilcoxon Mann-Whitney U tests were used to compare eNO, SpO2/FiO2 and eNO/ ( SpO2/FiO2 × 100) in two groups. Pearson's correlation analyses were used to determine the relationships between eNO levels and indices of hypoxemic respiratory failure. ResultsDuring the first two days of postnatal life, eNO values of HRF neonates were higher than those of the control neonates[day 1,(7. 9 ± 3.2 )× 10-9 vs. (5.8 ±1.8)×10-9, P＜0.05;day2, (8.8±3.2)×10-9vs. (6.0±2.4)×10-9, P＜0. 05], butthere were no significant differences in the following days. With SpO2/FiO2 increasing, difference of eNO values between the HRF and non-HRF controls became narrowed, but there was still two fold difference of eNO/(SpO2/FiO2 × 100) on day 5-7. ConclusionsA method for measuring eNO was established and there was difference in neonates with and without HRF, which diminished with prolonged postnatal days, reflecting pathophysiological characteristics of HRF.

Funds management,which has a direct effect on the development of scientific research projects,is an important part of the scientific research management in hospitals.By investigating the present situation of the scientific research funds management in a local hospital in Shanghai,this paper analyzes the problems commonly found in the scientific research funds management.Based on the findings of the analysis,this paper proposes some corresponding recommendations and countermeasures to improve the management.