Objective To study the mechanism of oxidative damage in myocardial tissue after limb ischemia reperfusion (IR), and the protective effects of heme oxygenase-1 on myocardial injury in experimental rats. Method The models of bilateral hind limbs ischemia and reperfusion in rats were established by using tourniquets applied to the roots of both hind limbs until palm blanched and pulseless for 4 hours. A total of 56 SD rats were randomly (random number) divided into 7 groups, namely one normal control group ( n = 8) and 6 ischemia-reperfusion groups as per different lengths of reperfusion time, e. g. 2 hrs, 4 hrs, 8 hrs, 16 h rs and 24 hr ( n = 8 each). The experimental rats were sacrificed after different lengths of reperfusion time. Specimens of myocardium and blood were taken for assays of malonaldehyde (MDA), superoxide dismutase (SOD) and myeloperoxidase (MPO), and pathological changes of myocardium were observed, and the expressions of HO-1 mRNA in myocardium were detected. Data were analyzed with ANOVA. Results (1) Compared with the control group, the levels of serum MDA and myocardial MDA of rats were increased in all IR groups and were higher (P < 0.05), and the levels of MDA reached the peak after reperfusion for 4 hours. The levels of serum SOD and myocardial SOD in rats of all IR groups were decreased and lower than those in rats of the control group ( P < 0.05), and the levels of serum SOD dropped away to the lowest point after reperfusion for 4 hours, and the levels of myocardial SOD fell off to the bottom after reperfusion for 8 hours. The levels of serum MPO and myocardial MPO were significantly increased in rats of all IR groups compared with the control group (P < 0.05). The levels of serum MPO reached peak after reperfusion for 4 hours, and the levels of myocardial MPO were increased to the highest spot after reperfusion for 6 hours. (2) The pathological changes in myocardium showed the most severe damage after reperfusionfor 4-6 hours.(3) After reperfusion for 2 hours, there were no significant differences in the expression of HO-1 mRNA between IR groups and control group (P >0.05), and after reperfusion for 4 hours and over, the expressions of HO-1 mRNA were markedly increased in IR groups and reached peak after reperfusion for 16 hours in comparison with the control group (P < 0.05). Conclusions The activation of neutrophils and free radicals may play a primarily adverse role in myocardial injury after limb IR, and the increase in the expression of HO-1 mRNA lessens the harm effects of IR on myocardium.

Objective To assess the value of chest computer tomography (CT) and serum tumor marker in lung cancer screening among health check-up populations.Methods A total of 88 596 adults who had a medical check-up (including chest CT,chest DR,and tumor biomarkers test) in our center were enrolled in this investigation.Clinical diagnosis of lung cancer was confirmed by three professional doctors.Receiver operating characteristic (ROC) curve was used to assess the relationship between serum level of tumor biomarkers and lung cancer.Results The average detection rate of lung cancer was 59.8/100 000 in our center during 2007 and 2011 (men 42.9/100 000,women 92.2/100 000).Rate of positive results of chest CT was 94.3％,which was approximately 2 times of chest DR.Moreover,carcinoembryonic antigen (CEA) testing was effective in early diagnosis of lung cancer with higher sensitivity (73.6％) and specificity (57.1％).Conclusion This study suggests that health checkup centers should pay more attention to lung cancer screening in high risk populations,and new technologies might be used to facilitate the early diagnosis of lung cancer.

Objective To analyze the effect of eukaryotic plasmids containing wild (sense) or anti- sense methylenetetrahydrofolate reductase (MTHFR) gene on cell viability and transcription level of tumor related genes in human gastric cancer cell line.Methods Human gastric cancer cell line MKN-45 was cultured.Recombinant plasmids containing wild MTHFR (W) or antisense MTHFR (A) gene, pCMV-W and pCMV-A,were constructed.Then pCMV-W,pCMV-A and pCMV blank plasmid were transfected into MKN45 cells respectively by using lipofect.Cell viability was analyzed by 3-(4,5-bime- thylthiazolyl-2)-2,5-diphenyhetrazolium dromide(MTT).The transcription levels of Dnmt 1,c-myc, p21~(WAF1) and hMLH1 genes were detected by real-time polymerase chain reaction(PCR).Results Cell vi- ability remarkably increased in those transfected with wild MTHFR (P＜0.01),which was contrary to those transfected with antisense MTHFR(P＜0.01).The expression of those tumor related genes mRNAs were all remarkably decreased in the MKN45-W cells in comparison with those in the MKN45-pCMV cells.No significant difference in the expressions of those tumor related genes mRNAs were found between the MKN45 cells transfected with pCMV-A and blank pCMV.Conclusion MTHFR influences cell viability and the expres- sion level of tumor related genes in human gastric cancer cell line MKN45.

In this study, the effect of heparin-derived oligosaccharide (HDO) on platelet-derived growth factor (PDGF) induced vascular smooth muscle cells (VSMCs) proliferation and the related signal transduction mechanisms were investigated. MTT assays were used to measure VSMCs proliferation. Cell cycle distribution was analyzed by flow cytometry. The level of key regulatory proteins in PKC, MAPK and Akt/PI3K pathways were determined by RT-PCR, Western blot and immunocytochemical methods. Meanwhile, mRNA expressions of some proto-oncogenes were assayed by RT-PCR method. Our data showed that HDO (0.01, 0.1 and 1 μmol · L(-1)) inhibited 30 ng · mL(-1) PDGF-induced VSMCs proliferation in a dose-dependent manner, blocked the G1/S transition and inhibited the level of key regulatory proteins and some proto-oncogenes (P < 0.05). The results showed that HDO may decrease the key regulatory proteins expression, hence suppress the transcription of proto-oncogene and G1/S transition, finally inhibiting VSMCs proliferation.
Cell Cycle ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Heparin ; pharmacology ; Humans ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Oligosaccharides ; pharmacology ; Platelet-Derived Growth Factor ; pharmacology ; Signal Transduction

Objective To investigate the effects of erythropoietin (EPO) pretreatment on the acute lung injury induced by lipopolysaccharide (LPS) in rats and the underlying mechanism. Methods Thirty-two male SD rats weighing 180-220 g were randomly divided into 4 groups (n=8 each): group Ⅰ control (C);group Ⅱ EPO;group Ⅲ LPS and group Ⅳ EPO + LPS. EPO 3 000 U/kg was given IP in group Ⅱ , LPS 6 mg/kg was given iv in group Ⅲ . In group Ⅳ EPO 3 000 U/kg was given IP at 30 rain before iv LPS 6 mg/kg, The animals were killed at 4 h after LPS administration. Lung tissue specimens were obtained for microscopic examination. Wet/dry ratio (W/D), myeloperoxidase (MPO) activity, malondialdehyde (MDA) and nitric oxide (NO) content in lung tissue were determined. The expression of inducible nitric oxide synthase (iNOS) and nitrotyrosine (NT) in lung tissue was determined by Western blot. Results W/D ratio, MPO activity, MDA and NO content were significantly increased and iNOS and NT expression was significantly up-regulated in LPS group as compared with control group. EPO pretreatment significantly attenuated the LPS-induced changes in group EPO + LPS. Conclusion EPO pretreatment can ameliorate the acute lung injury induced by LPS by down-regulating iNOS expression and reducing NO production.

OBJECTIVETo investigate the semen quality and its influencing factors in preconception males in Nanjing area so as to provide some evidence for working out effective intervention measures.

METHODSTotally 687 men receiving preconceptional physical examination were enrolled in this study. A questionnaire survey was conducted among the subjects along with an analysis of their semen quality.

RESULTSThe median of sperm concentration was 63.3 x 10(6)/ml (95% CI [19.88-119] x 10(6)/ml). The median of grade a sperm was 33.03% (95% CI [19.38-55.05]%), that of grade a + b sperm was 52.08% (95% CI [39.53-69.37]%), and that of teratosperm was 91.75% (95% CI [69-100]%). The median concentration of seminal plasma PMN-elastase was 195.55 ng/ml (95% CI [76.16-3330.38] ng/ml) and that of seminal plasma zinc was 7.62 μmol/L (95% CI [1.5-23, 45] μmol/L). The positive rates of Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), and Gardnerella vaginalis (GV) were 42.4%, 0.3%, and 2.4%, respectively. The median of sperm DNA fragmentation index (DFI) of those whose wives had a history of adverse pregnancy was 20.25% (95% CI [2.15-68.25]%). Multivariate logistic regression analysis suggested that mental stress (OR 1.567, 95% CI [1.081-2.27]) and sedentariness (OR 1.772, 95% CI [1.211-2.592]) were independent risk factors for asthenospermia.

CONCLUSIONThe sperm quality of preconception males in Nanjing area is not encouraging, and it can be improved by changing undesirable lifestyle and reducing mental stress.

Adult ; Asthenozoospermia ; etiology ; China ; Chlamydia trachomatis ; isolation & purification ; DNA Fragmentation ; Gardnerella vaginalis ; isolation & purification ; Humans ; Leukocyte Elastase ; analysis ; Male ; Preconception Care ; Semen ; microbiology ; Semen Analysis ; statistics & numerical data ; Sperm Count ; statistics & numerical data ; Spermatozoa ; Ureaplasma urealyticum ; isolation & purification

Objective: To investigate the role of human umbilical vein endothelial cells (HUVEC) in supporting hematopoiesis and the expansion of hematopoietic stem/progenitor cells in vitro. Methods: According to the fact that HUVEC supernatant has colony stimulating activity shown by methylcellulose colony-forming assay and HUVEC can maintain the survival of mononuclear cells for at least four weeks in vitro, CD34+ cells from umbilical cord blood were seeded with (HUVEC group) or without (control group) HUVEC monolayer. Every week cells were collected and counted, the frequency of CFU-GM was measured by using methylcellulose colony-forming assay, and the percentage of CD34+ and CD41a+ cells was measured by flow cytometry. Results: In control group，all the CD34+ cells died in two weeks. However, in HUVEC group,most nucleated cells and CD34+ cells were expanded by 68.1±14.8 fold and 6.6±1.4 fold，respectively at the third week while CFU-GM expansion reached its peak (5.7±2.1 fold) at the week 2. Moreover, the percentage of CD41a+ cells was enhanced significantly, reaching a maximum (15.6%) at the week 3. Conclusions:HUVEC can support hematopoiesis in vitro and expand the hematopoietic progenitor cells and CD41a+ cells in direct contact coculture.

Objective: To establish an adult rat model of lung blast injury in laboratory for providing a reliable rat model for the diagnosis and treatment of lung blast injury. Methods: According to the different injury driving pressure of BST-I biological shock tube, 40 adult SD male rats aged 7 to 8 weeks were randomly divided into 4 groups (10 each): 4.5 MPa group, 4.8 MPa group, 5.0 MPa group and 5.5 MPa group. The general physiological and 24-hour survival status of rats in each group were recorded before and after modeling. The survived rats 24 h after modeling were anesthetized and then killed, and the chest anatomy was performed. The characteristics of lung injury were observed, the degree of lung tissue injury was graded using organ injury scaling (OIS) and abbreviated injury scale (AIS). Furthermore, the left lower part of lung tissue was taken for routine pathological section, and the degree of lung tissue injury was observed under light microscope. Results: All rats were successfully modeled and survived 24 h after injury, and showed shortness of breath and accelerated heart rate without hematuria and blood stool. There were significant differences among the four groups in bleeding rate of external auditory canal, anal temperature and pulmonary bleeding area (P<0.01). The grading scores of lung injury degree in four groups were as follows: for 4.5 MPa group, the OIS grade concentrated in grade II, the AIS score concentrated in score 1; for 4.8 MPa group, the OIS grade concentrated in grade HI, the AIS score concentrated in score 2; for 5.0 MPa group, the OIS grade concentrated in grade IV, the AIS score concentrated in score 3; for 5.5 MPa group, the OIS grade concentrated in grade V, and the AIS score concentrated in score 4. There existed significant differences in grading and score among the four groups (P<0.05). However, the results of OIS classification showed that the lung injury grades of 5.0 MPa group and 5.5 MPa group were higher than that of 4.5 MPa group, and the grade of 5.5 MPa group was higher than that of 4.8 MPa group. As to AIS score, statistical differences existed between any two groups on the lung tissue injury score (P<0.05), and the AIS scores ranged from high to low were as follows: 5.5 MPa>5.0 MPa>4.8 MPa>4.5MPa. Under light microscope, rats in the four groups showed varying degrees of pulmonary hemorrhage, edema and atelectasis, and the lung tissue was ruptured. Conclusion: The stable adult SD rat models of mild, moderate, severe and extremely severe lung blast injury have been established.

A boy, aged 66 days, was admitted to the hospital due to subcutaneous nodules for 46 days and abdominal distension for 10 days. The main clinical manifestations were loss of adipose tissue, subcutaneous nodules, insulin-resistant diabetes, hypertriglyceridemia, and hepatic steatosis. The boy was diagnosed with congenital generalized lipodystrophy type 1 (CGL1). His condition was improved after administration of middle-chain fatty acid formula milk and insulin injection or oral metformin. Gene testing revealed a homozygous mutation, c.646A>T, in the AGPAT2 gene, and both his parents were carriers of this mutation. This case of CGL1 has the youngest age of onset ever reported in China and multiple subcutaneous nodules as the initial symptom.
Adipose Tissue ; China ; Fatty Liver ; Humans ; Infant ; Insulin Resistance ; Lipodystrophy ; Lipodystrophy, Congenital Generalized ; Male

Objective To evaluate the rationality of predisone dosage in the treatement of SLE with different clinical scoring systems. Methods The clinical data of 51 newly diagnosed patients with systemic lupus erythematosus(SLE) were collected, and disease activity was assessed by SLEDAI- 2000, BILAG- 2004 and SLAM-R scoring systems. The correlations between SLEDAI- 2000, BILAG- 2004 , SLAM-R scores and prednisone dosage were analyzed. Results BILAG-2004 score showed the best correlation with the dosage of prednision (r=0.827, P<0.001) .SLAM score showed the worst correlation with the dosage of prednision (r=0.512, P<0.001). Different treatment choice was associated with different organ/system involvement. Conclusions BILAG- 2004 , SLEDAI- 2000 and SLAM-R scoring systems all can be used as references for the prednisone dosage slection.It is suggested that the BILAG- 2004 scoring system is more reliable and comprehensive in clinical practice．