BACKGROUND:Cardiopulmonary exercise test plays a significant role in the evaluation of cardiopulmonary function, but it is limited by expensive equipments and professional personnel, and moreover, the subjects need to bear the maximal exercise intensity. As a result, it is extremely urgent to find a submaximal exercise test characterized by simple operation, low cost and easy to be popularized and used. OBJECTIVE:To compare the maximal oxygen uptake in cardiopulmonary exercise test and 6-minute stair climbing and descending test. METHODS:Sixty-seven volunteers were recruited to undergo the cardiopulmonary exercise test using the Bruce protocol, and then, the maximal oxygen uptake was detected. After that, al the participants were subjected to 6-minute stair climbing and descending test, folowed by determination of the maximal oxygen uptake. RESULTS AND CONCLUSION:The maximal oxygen uptake in the cardiopulmonary exercise test was significantly higher than that in the 6-minute stair climbing and descending test (P < 0.01), and there was a highly positive correlation and consistency between the maximal oxygen uptakes in the two tests (r=0.911,P < 0.01). Therefore, 6-minute stair climbing and descending test can be used to detect the maximal oxygen uptake, which may become an effective method for evaluating cardiopulmonary function.

Objective To observe the effects of high fat diet and 6?week voluntary wheel running on the structure of hypothalamus, pituitary and testis and sex hormone levels in pubertal male rats. Methods Forty 3?week?old male Spra?gue?Dawley rats were randomly divided into following groups: the control group fed with normal diet (C), training group fed with normal diet ( CE) , control group fed with high fat diet ( D) , and training group fed with high fat diet ( DE) , 10 rats in each group. The groups C and D were bred for 8 weeks freely, and the groups CE and DE were assigned to have vol?untary wheel running twice/d, 1 h/time, 5 d/w, for 6 weeks after a 2?week adaptive feeding. Eight weeks later, blood sample was collected to detect the serum T, E2 , FSH and LH, and the E2 and T of testis were also detected. The histology of hypothalamic, pituitary, and testis tissues was observed by light microscopy. Results ( l) Compared with the group C, the group D had significantly decreased levels of serum T and E2 and testicular T (P<0?05), and significantly increased serum E2, FSH and LH levels (P<0?05). In the group D, vacuolar lipid droplets were increased in the hypothalamus, e?osinophils and basophils were reduced in the pituitary, and the area of seminiferous tubules, percentage of sperm cells, and quantity of Leygid cells were significantly decreased in the testis. (2) Compared with the group D, the serum T and testic?ular T concentrations were increased, but the serum E2, FSH and LH were decreased significantly (P<0?05) in the group DE, and vacuolar lipid drops were increased in the hypothalamus, eosinophil cells were increased in the pituitary gland, and the area of seminiferous tubules was increased but not significantly in the testis. Conclusions 6?week voluntary wheel exercise can improve the high fat diet?induced abnormal secretion of sex hormones, but not effectively improve the histologi?cal changes in hypothalamus, pituitary gland and testis.

AIM: To study the changes in myocardial blood flow (MBF), capillarization and cardiac function in the rat during acclimation to hypoxia. METHODS: Myocardial capillary density (CD) and capillary/myocyte ratio (C/M) was assayed by alkaline phosphatase histochemistry. Biomicrosphere method was used to determine MBF in the rat after 5, 15 or 30 days hypobaric hypoxic exposure (5 000 m). RESULTS: In the course of hypoxia, MBF and cardiac function increased in the right ventricle. However, in the left ventricle, acute hypoxia caused an increase in MBF and a decrease in cardiac function. Both returned to the control level on continued hypoxic exposure. Neovascularization occurred after 15 day or 30 day of hypoxic exposure in both ventricles, judged from the significant increment of C/M ratio albeit the CD remained unchanged in the right ventricle. CONCLUSION: Our findings indicate that adaptive changes in rat heart during acclimation to hypoxia include: ① persistent increase in MBF, hypertrophy associated with increase in capillarity and enhanced cardiac function of the right ventricle; ② increase in MBF and depression of cardiac function at first, then followed by recovery of MBF and increase in capillarity accompanied with recovery of left ventricular function.

AIM: To compare the effects of hypoxia on expression of vascular endothelial growth factor (VEGF), iNOS and eNOS mRNA in cultured umbilical vein endothelial cells (UVECs) obtained from Tibetan and Han. METHODS: UVECs were obtained from native Tibetan and immigrant Han, respectively and cultured under hypoxia conditions (0.5% oxygen) for 2 h, 4 h, 12 h, and 24 h and normoxic conditions. VEGF, iNOS and eNOS mRNAs were detected with methods of RT-PCT. RESULTS: VEGF and iNOS mRNAs were up-regulated while eNOS mRNA depressed by hypoxia similarly in Tibetan and Han UVECs. CONCLUSION: Our results suggest that the changes of VEGF, iNOS and eNOS mRNA expression are common pathways in the mechanisms of hypoxic responses.

Objective:To evaluate the clinical application value of gastrointestinal contrast-enhanced ultrasound combined gas injection method in verifying the location of nasointestinal tube in critically ill patients.Methods:Data of 60 critically ill patients who had the indications of indwelling nasointestinal tube were collected from September 1,2015 to September 1,2016 in the Intensive Care Unit of Zhejiang Provincial People(s) Hospital.The position of nasointestinal tube in patients who underwent bedside blind insertion would be confirmed routinely through gas injection auscultation method.After tube was inserted,its route was scanned by ultrasound with gas perfusion assistance.Afterwards,rapid gas perfusion was used until suspicious tube end position was determined.Furthermore,oral ultrasound contrast agent was injected into the tube if instantaneous strong echo of gas was observed in localized lumen,and contrast agent filling meant the placement being successful.Two methods of position confirmation of nasointestinal tube in critically ill patients included gastrointestinal contrast enhanced ultrasound combined gas injection and gas injection auscultation only,and the effect of the two methods was compared and confirmed by chest and abdominal X ray examinations to verify the location of nasointestinal tube below pylorus.Results:A total of 60 patients were included in this study,58 patients(96.7％)in gastrointestinal contrast enhanced ultrasound combined gas injection group were successfully positioned.Among them,the placements of tube in 56 cases were below pylorus,while 2 cases were above pylorus.The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of location of gastrointestinal contrast enhanced ultrasound combined gas injection method were 96.6％,100％,100％,50％,96.7％ and of gas injection auscultation method were 74.1％,50％,97.7％,6.3％ and 73.3％.The differences of the sensitivity,specificity,negative predictive value and accuracy between the two methods were statistically significant (P ＜ 0.05).Conclusion:Gastrointestinal contrastenhanced ultrasound combined gas injection method is a safe,simple and convenient method with high sen-sitivity,specificity,negative predictive value and accuracy in confirming the location of the nasointestinal tube.

Objective　To investigate morphological changes of endothelial cells after nordihydroguaiaretic acid (NDGA) treatment in vitro. Methods　The morphological changes of human umbilical vein endothelial cells (HUVEC) cell line ECV-304 and the cell apoptosis rate in sub-G0 phase were observed with invert, light and electron microscope and flow cytometry after NDGA treatment at different concentrations or with PBS (0.01 mol/L) as control. Results　①After the treatment of NDGA at 50～200 μmol/L for 1～3 d or up to 8 d at 100 μmol/L, ECV-304 cells tended to elongate into a shuttle-like sparse appearance and those in mitosis were decreased, indicating the suppression of cell proliferation. All these alteration was in a time-and dose-dependent manner. ②NDGA-treated ECV-304 cells displayed morphological features of apoptosis, especially at the 48th h after the treatment. With flow cytometry, the cells in sub-G0 phase were significantly increased, and reached its peak at hour 12 (20.42%) after NDGA treatment. In addition, the degeneration and necrosis of ECV-304 cells were related to the concentrations of NDGA. Conclusion　NDGA can inhibit the proliferation and growth of endothhelial cells, and induce apopotosis, which might also inhibit angiogenesis.

Objective　To explore the effect of nordihydroguaiaretic acid (NDGA) on the expressions of vascular endothelial growth factor (VEGF) and its receptor, kinase-inserted domain containing receptor(KDR) and the possible mechanism. Methods　The expression of VEGF in human malignant glioma cell line SHG-44 and that of KDR in human umbilical vein endothelial cell (HUVEC) line ECV-304 were observed 1～3 d after NDGA treatment with immunohistochemistry, in situ hybridization and image analysis. Results　The expression of VEGF was declined at protein or mRNA levels in SHG-44 cells after treated with 100 μmol/L NDGA for 1 to 3 d. The expression of KDR in endothelial cells with 100 μmol/L NDGA treatment for 1 to 3 d was decreased too, in a more obvious way compared with the decline of VEGF expression in SHG-44 cells. Conclusion　The results suggest that NDGA inhibits the expression of VEGF in glioma cells as well as that of VEGF receptor KDR in endothelial cells, which may be the important molecular mechanism of anti-angiogenesis of NDGA.

We stimulated preadipocyte of mice with calcium acetate, p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580, the paralysors and excitomotors of calcium channel. Then we detected expression level of preadipocyte differentiation's marker genes and calcium signal related acceptor genes by real-time PCR, and determined intracellular free Ca2+ concentration ([Ca2+]i]) with Fura-2/AM, intracellular lipid accumulation by oil red O staining. Our aim was to investigate the potential mechanism between calcium signal and preadipocyte differentiation. The results indicated that the paralysors and excitomotors of calcium channel changed the expression level of lipoprotein lipase (LPL), peroxisome proliferators-activated receptor gamma (PPARgamma), fatty acid synthetase (FAS), and the lipid accumulation, markedly. Compared with exocellular Ca2+'s decrease, inhibited intracellular Ca2+'s liberation can promoted preadipocyte differentiation (P < 0.01), and compared with intracellular Ca2+'s increase, promoted exocellular Ca2+'s ingest inhibited preadipocyte differentiation (P < 0.01). SB203580 degraded [Ca2+]i, promoted differentiation marker genes' expression and lipid accumulation in preadipocyte (P < 0.01). But calcium signal didn't have effects to vitamin D receptor (VDR) and extracellular Ca2+-sensing receptor (CaSR)'s expression. It indicated that calcium signal may effect preadipocyte different and lipid accumulation by p38 MAPK pathway.
Adipocytes ; cytology ; Animals ; Calcium ; metabolism ; Calcium Signaling ; drug effects ; Cell Differentiation ; physiology ; Cells, Cultured ; Imidazoles ; pharmacology ; Lipids ; biosynthesis ; Mice ; Pyridines ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

Objective:To investigate the role of LncRNA-TUG1 in the injury of intestinal epithelial cells induced by lipopolysaccharide (LPS).Methods:LPS was used to treat HIEC-6 human intestinal epithelial cells for 24 h to construct a sepsis injury model. Whole transcriptome RNA sequencing was used to analyze the expression changes of mRNA, microRNA and lncRNA in HIEC-6 cells after LPS treatment. Real-time fluorescence quantitative (qRT-PCR) and Western blot was performed to detect the expression changes of lncRNA-TUG1, microRNA-132-3p (miR-132-3p), SIRT1 mRNA and SIRT1 protein in HIEC-6 cells after LPS treatment. The expression levels of LncRNA-TUG1, miR-132-3p and SIRT1 were artificially changed by in vitro transfection. qRT-PCR and Western blot were used to confirm the regulatory effect of lncRNA-TUG1 on microRNA-132-3p and SIRT1. CCK-8 and flow cytometry were used to analyze the effects of LncRNA-TUG1, miR-132-3p and SIRT1 on the proliferation and apoptosis of HIEC-6 cells. The dual luciferase report analysis was used to verify the targeting relationship between LncRNA-TUG1, miR-132-3p and SIRT1. Statistical analysis was performed using SPSS 17.0, and differences between the two groups were compared using independent sample t test. Results:RNA sequencing results showed that the expressions of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells after LPS treatment ( t=3.26, P<0.05 and t=2.55, P<0.05), but the expression of miR-132-3p was increased ( t=4.12, P<0.05). In vitro cell experiments, the expression of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells treated with LPS ( t=5.69, P<0.05 and t=5.712, P<0.05), while the expression of miR-132-3p was increased ( t=3.88, P<0.05). Overexpression of lncRNA-TUG1 increased the proliferation rate ( t=6.55, P<0.05) and decreased the apoptosis rate ( t=3.94, P<0.05) of LPS-treated cells. Upregulation of lncRNA-TUG1 decreased the expression of miR-132-3p ( t=4.66, P<0.05), and increased the mRNA and protein levels of SIRT1 ( t=3.91, P<0.05). Transfection of miR-132-3P mimic could inhibit the mRNA ( t=4.08, P<0.05) and protein levels of SIRT1. In LPS-treated cells, the cells co-transfected with miR-132-3pmimic and siRNA-SIRT1 had a lower proliferation rate ( t=4.55, P<0.05 and t=5.67, P<0.05) and a higher apoptosis rate ( t=3.90, P<0.05 and t=4.22, P<0.05) than those transfected with only pcDNA3.1-lncRNA-TUG. Conclusions:lncRNA-TUG1 may act as a ceRNA to regulate miR-132-3p/SIRT1, therefore alleviating HIEC-6 cell injury caused by LPS. Intervention of lncRNA-TUG1/miR-132-3p/SIRT1 regulatory pathway may become a potential strategy to prevent sepsis-induced intestinal mucosal damage.

Objective To investigate the prevalence of feeding intolerance (FI),and to explore the FI within 7 days of ICU admission in association with clinical outcome in critically ill patients.Methods The adult patients from 14 general ICUs in Zhejiang Province with an expected admission to ICU for at least 24h were recruited from March 2014 to August 2014,and all clinical,laboratory,and survival data were prospectively collected.The AGI (acute gastrointestinal injury) grade was daily assessed based on gastrointestinal (GI) symptoms,feeding details and organ dysfunction within the first week of ICU stay.The intra-abdominal pressures (IAP) was measured using AbViser device.Results Of 550 patients enrolled,418 were assessed in GI symptoms and feeding details within 7 days of ICU stay.The mean age and SOFA score were (65.1 ± 18.3) years and (8.96 ±4.10),respectively.Of them,355 patients (84.9％) were under mechanical ventilation support,and 37 (8.85％) received renal replacement therapy.The mean length of time for enteral feeding was (30.8 ±26.2) h,and the prevalence of FI on the 3rd and 7th day of ICU stay accounted for 39.2％ and 25.4％,respectively.Compared to those with FI within 7 days of ICU stay,the patients without FI had higher rate of successively weaning from mechanical ventilation (21.3％ vs.5.7％,P =0.003) and higher rate of withdrawal of vasoactive medication (45.5％ vs.20.0％,P =0.037),as well as lower mortality rate of 28-day (24.4％ vs.38.7％,P =0.004) and 60-day (29.6％ vs.44.3％,P =0.005).In multivariate Cox regression model with adjustment for age,sex,participant center,serum creatinine and lactate,AGI grade on the first day of ICU stay,and comorbidities,the FI within 7 days of ICU stay (x2 ≥ 7.24,P ＜ 0.01) remained to be independent predictors for 60-day mortality.After further adjusted for SOFA score,the FI within 7 days of ICU stay (HR =1.71,95％ CI:1.18-2.49;P =0.006) and AGI grade on the first day of ICU stay (HR =1.33,95 ％ CI:1.07-1.65;P =0.009) could provide independent prognostic values of 60-day mortality.Conclusions There is high rate of FI occurred within 7 days of ICU stay,and is significantly associated with worse outcome.In addition,this study also provides evidence to further support that measurement of gastrointestinal dysfunction could increase value of SOFA score in outcome prediction for the risk of 60-day mortality.