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Objective To explore the effect of transforming growth factor β1 (TGF-β1) on epithelial-mesenchymal transition in rat peritoneal mesothelial cells(RPMCs) and its mechanism.Methods Primary peritoneal mesothelial cells of SP rats were cultured in vitro. After synchronization for 24 h, RPMCs were randomly divided into 2 groups: Group A (control), Group B (TGF-β1, 10 μg/L). RPMCs were stimulated by 10 μg/L TGF-β1 for different time. The mRNA and protein expression levels of E-cadherin, α-smooth muscle actin (α-SMA) and collagenⅠwere measured by RT-PCR and Western blot, respectively. The protein expression level of total RhoA was measured by Western blot. Active RhoA was extracted by Plasma Membrane Protein Extraction Kit, and assessed by Western blot. Results TGF-β1 down-regulated mRNA and protein expression of E-cadherin in RPMCs, and upregulated mRNA and protein expression of α-SMA and CollagenⅠ. TGF-β1 stimulation elicited a robust increase in RhoA activity in a time-dependent manner. RhoA activity peaked at 1 h.Conclusion RPMCs can be transdifferentiated into myofibroblast under the effect of TGF-(β1,)and the mechanism may be related to the activation of RhoA associated signal pathway.
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Objective To investigate the role of RhoA-Rock signaling pathway in the process of rat peritoneal mesothelial cells (RPMCs) epithelial-mesenchymal transition (EMT) induced by transforming growth factor β1 (TGF-β1). Methods Primary RPMCs were cultured in vitro. After synchronization for 24 hours, RPMCs were randomly assigned to 4 groups: group A (control), group B (TGF-β1, 10 μg/L), group C (10 μg/L TGF-β1+10 μmol/L Y-27632, an inhibitor of Rock, pretreated for 2 hours with Y-27632 before TGF-β1 stimulation), group D (Y-27632 alone, 10 μmol/L). Growth arrested and synchronized RPMCs were stimulated by 10 μg/L TGF-β1 for different time. The mRNA and protein expression levels of E-eadherin, α-SMA and collagen Ⅰ were measured by RT-PCR and Western blotting respectively. The protein expression level of vimentin was measured by Western blotting. Active RhoA was extracted by Plasma Membrane Protein Extraction Kit, then it was assessed by Western blotting. Results (1) TGF-β1 stimulation elicited a robust increase in RhoA activity in time-dependent manner, which was (2.57±0.52) folds compared with control group (P<0.05) after 10 min stimulation. RhoA activity peaked at 1 hour, which was (4.35±0.41) folds compared with control group (P<0.05). (2) TGF-β1 up-regulated mRNA and/or protein expression of α-SMA, vimentin and collagen Ⅰ , and down-regnlated mRNA and protein expression of E-cadherin in RPMCs. (3) The Rock inhibitor Y-27632 effectively revered TGF-β1-induced expression of α-SMA, collagen Ⅰ and vimentin. The mRNA levels of α-SMA and collagen Ⅰ decreased by 53.8% and 55.7%, and the protein levels of α-SMA, vimentin and collagen Ⅰ decreased by 42.6%, 60.1% and 58.1% compared with TGF-β1-stimulated groups (P< 0.05). But Y-27632 had no effect on the level of E-cadherin. Conclusions RhoA-Bock signaling pathway may mediate EMT induced by TGF-β1 in rat peritoneal mesothelial cells. RhoA-Rock pathway may be the potential therapeutic target in the progress of peritoneal fibrosis.
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BACKGROUND: Studies have confirmed that RhoA/ROCK signaling pathway plays an important role in the process of epithelial-mesenchymal transition. OBJECTIVE: To investigate the effect of Y-27632 (inhibition of rhodopsin kinase of 4-aminopyridines compounds) on the epithelial-mesenchymal transition of rat peritoneal mesothelial cells (RPMCs) induced by transforming growth factor ?1 (TGF-?1). DESIGN,TIME AND SETTING: An animal observational experiment was performed at the Laboratory of Nephrology in the First Affiliated Hospital of Sun Yat-sen University from March 2007 to March 2008. MATERIALS: A total of ten healthy male Sprague Dawley rats were provided by the Animal Experimental Center of Sun Yat-sen University. Y-27632 was the product of Calbiochem Company (German). TGF-?1 was the product of R&D Company (USA). METHODS: Primary RPMCs were cultured in vitro. After synchronization for 24 hours, RPMCs were randomly divided into 4 groups: control group (RPMCs cultured in the DMEM/F12 without serum), TGF-?1 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L TGF-?1), Y-27632 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L Y-27632), TGF-?1 +Y-27632 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L Y-27632. 2 hours later, 10 ?g/L TGF-?1 were added). The cells were collected 48 hours after culture. MAIN OUTCOME MEASURES: The mRNA and protein expression of E-cadherin and ?-Smooth muscle actin (?-SMA) were measured by RT-PCR. The protein expression level of E-cadherin, ?-SMA and vimentin was measured by Western blotting. RESULTS: Compared to control group, the expression of E-cadherin significantly decreased in the TGF-?1 group, and the expression of ?-SMA and vimentin significantly increased (P
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OBJECTIVE To study the epidemiological features of hospital lower respiratory tract infection in order to adopt effective control methods.METHODS We investigated and analyzed the 45 patients suffered from hospital lower respiratory tract infections among the 9 449 hospitalizd patients between Jan 2000 and Jun 2002.RESULTS The rate of total hospital lower respiratory tract infection was 0.48%,the composition rate was 23.56%,the average bacteria resistance rate was 57.17%.CONCLUSIONS In order to prevent hospital lower respiratory tract infection,immune support therapy should be strengthened and risk factors should also be controlled.
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BACKGROUND: In the past, a lot of researches used one-time lead level in umbilical cord blood at birth for investigations,however, one-time lead level in umbilical cord blood at birth can not represent lead caused cumu lative injury to neonatal nervous system during the whole period of preg nancy. Lead in meconium is mainly from digestive juice secreted by fetal alimentary tract, exfoliative epithelia from neonatal intestinal tract and am niotic fluid and sebum cutaneum swallowed by fetus, which is excreted from the very start of pregnancy to 24 hours after birth of neonates and re flects the lead deposit in neonatal intestinal tract during the whole period of pregnancy. OBJECTIVE: To explore the relationship between neurobehavioral devel opment in neonates with intrauterine exposure to lead at low level and the lead level in umbilical cord blood (CBPb) and meconium (MPb). DESIGN: Take lead levels in umbilical cord blood and meconium as neonatal intrauterine exposure indicators and scores of neonatal neurobe havioral development as effect indicators, and descriptive analysis is used to evaluate the correlativity. SETTING: Wuhan University of Science and Technology; Laboratory of Occupation Disease and Epidemiology, Tongji Medical College, Huazhong University of Science and Technology; Wuhan First Metallurgical Con struction Company Hospital for Workers and Staff. PARTICIPANTS: A total of 103 cases of full-term and healthy neonates were selected as objects of observation. The neonates were born in Depart ment of Gynecology and Obstetrics, Wuhan First Metallurgical Constrction Company Hospital, Qingshan District of New Industrial District of Wuhan from January to October 1999. Their parents were agreed to participate in the study and filled in the questionnaire, and provided neonatal meconium and performed neonatal tests on schedule. METHODS: ① Collection and assay of sample: 5 mL umbilical cord blood were collected and reserved in refrigerator at -4 ℃. Meconium with in 24 hours after birth, with dry weight between 5 to 10 g was collected, the lead levels in umbilical cord blood and meconium were assayed with the method of graphite furnace atomic absorption spectroscopy. ② Group ing: The neonates were divided into two groups with high and low-exposure to lead based on the cutoff value of CBPb of 0.483 μmol/L and MPb of 127.78 mg/kg. ③ Neonatal neurobehavioral development examination: Neonatal neurobehavioral development examination method was used for examination 3 days before delivery. Meanwhile, self-designed questionnaire was used to conduct a survey in puerperas. MAIN OUTCOME MEASURES: ① Lead levels in neonatal umbilical cord blood and meconium. ② Scores of neurobehavioral development of neonates with different lead levels of umbilical cord blood and meconium. RESULTS: All the 103 cases of neonates entered results analysis. ①There was significant difference only in scores of neonatal behavioral neurological assessment (NBNA) and biological visual and auditory orientation reaction (BVAOR)between groups with high and low-exposure to lead in umbilical cord blood (P ＜ 0.05). However, there was no rank correlativity between lead level in umbilical cord blood and scores of NBNA,non-biological auditory orientation reaction (NBAOR), non-biological visual orientation reaction (NBVOR) and biological visual and auditory orientation reaction (NBVOR). ②here was significant difference in scores of NBNA,NBAOR, NBVOR and BVAOR between groups with high and low-exposure to lead in meconium (P ＜ 0.05-0.01). The lead level in meconium clearly correlated reversely with scores of NBNA, NBVOR and BVAOR.CONCLUSION: Lead level in meconium is more sensitively related to the scores of neonatal neurobehavioral development, which could be used as indicator for lead deposit in the fetal body during the period of pregnancy.
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BACKGROUND: Logistic analysis of multi-ordered response-variable is used to probe into from another view the interrelationship between lead content in fetus faeces and neurobehavioral development under exposure to low-level lead in uterus.OBJECTIVE: To probe into multi-factors of neurobehavioral development in neonates and the sensibility.DESIGN: The total score of neurobehavior in neonates was taken as dependent variable and 24 indexes were as independent variables, such as induced factors, lead in umbilicus blood and lead in fetus faeces in questionnaire. Logistic progressive regression of multi-ordered response-variables was used in analysis and corresponding factors were screened at level of P=0.10.SETTING: Wuhan University of Science and Technology, Laboratory Room of Occupation Disease and Epidemic disease in Tongji Medical College of Huazhong University of Science and Technology and Occupation Hospital of China First Metallurgical Construction General Company.PARTICIPANTS: Totally 103 full-month borne and healthy neonates were randomized in Department of Gynecology of one occupation hospital in Qingshan District of New-type Industry Area of Huanhan City from January to October 1999 as the objects. The relatives agreed with topic research and questionnaire investigation and they provided neonatal faeces and received neonatal tests on time.off the umbilicus and preserved in freezing in refrigerator at -4 ℃. The faeces in 24 hours after birth was collected and the lead contents of umbilicus blood and faeces were assayed with graphite furnace atomic absorpBehavioral Neurological Assessment (NBNA) was used in examination on the 3rd day after delivery. Simultaneously, the self-designed questionnaire was adopted in the investigation for parturients. The questionnaire involved other possible factors of neonatal neurobehavioral development, including dependent variables, concerning to states of family, society, environment and health that affected neurological development in neonates and lead contents in umbilicus blood and faeces. Scores of neonatal neurobehavior were taken as response variables. Finally, the corresponding factors were screened.MAIN OUTCOME MEASURES: To screen the factors of neurobehavioral development of neonates.RESULTS: Totally 103 cases entered result analysis. Six factors were selected in the model, named pregnant weeks, the month of drug administration in pregnancy, hemoglobin, emotions in pregnancy, lead level in fetus faeces and drug administration.CONCLUSION: Neonatal neurobehavioral development was related to multiple factors. Good nutrient in pregnancy, long pregnant weeks and good emotions in pregnancy benefit neurobehavioral development of neonates. Drug administration during pregnancy is disadvantageous in neonatal neurobehavioral development, especially the medication at the early phase of pregnancy. The increased lead content in neonatal faeces does not benefit neonatal neurobehavioral development.