Cigarette smoke is associated with the development of several diseases, such as chronic ob- structive pulmonary disease （COPD）. The purpose of this study was to investigate genotoxicity and heat shock protein 70 （H...Cigarette smoke is associated with the development of several diseases, such as chronic ob- structive pulmonary disease （COPD）. The purpose of this study was to investigate genotoxicity and heat shock protein 70 （Hsp70） in human airway smooth muscle cells （HASMCs） exposed to cigarette smoke extract （CSE）. HASMCs was exposed to CSE with different doses for 24 h. The level of 8-hydroxydeoxyguanosine （8-OHdG） was determined by using HPLC-ECD, the DNA damage was ana- lyzed by using comet assay, and apoptosis was examined by using Annexin-FITC/PI staining. The pro- duction of Hsp70 after CSE stimulation was tested. Results indicated that CSE significantly increased the level of 8-OHdG, DNA damage and cell apoptosis, and reduced the production of Hsp70. In par- ticular, levels of Hsp70 were inversely correlated with 8-OHdG, DNA damage and cell apoptosis. It was concluded that cigarette smoke induced genotoxicity and decreased the production of cell protective protein Hsp70, which may contribute to the development of some airway diseases.展开更多
The effects of cigarette smoke extract (CSE) on the expression of heat stress protein 70 (Hsp70) in human bronchi smooth muscle cells were investigated in vitro, and the changes in Hsp70 mRNA in the patients with chro...The effects of cigarette smoke extract (CSE) on the expression of heat stress protein 70 (Hsp70) in human bronchi smooth muscle cells were investigated in vitro, and the changes in Hsp70 mRNA in the patients with chronic obstructive pulmonary disease and their significance were explored. Human bronchi smooth muscle cells were cultured with CSE at the different concentrations. The expression of Hsp70 mRNA and Hsp70 was detected by reverse translation-polymerase chain reaction (RT-PCR) and Western blotting respectively. Levels of Hsp70 mRNA and Hsp70 in lymphocytes from 20 patients with COPD and 20 healthy smoking control subjects were measured by RT-PCR and Western blotting. The results showed the expression of both Hsp70 mRNA and Hsp70 was decreased conformably in human bronchi smooth muscle cells treated with CSE at certain concentration in vitro. The A values of the Hsp70 mRNA expression were 0.24±0.11 and 0.42±0.13 respectively in COPD patients and healthy smoking controls with the difference being significant (P<0.01). There was also significant difference in the A values of the Hsp70 expression between COPD patients and healthy smoking controls (20.9±9.9 vs 44.8±15.3, P<0.01). The levels of Hsp70 mRNA had strongly positive correlation with Hsp70 protein (r = 0.85, P<0 01). It was suggested that the expression of Hsp70 mRNA was in concordance with the expression of Hsp70, which could provide a basis on the study of Hsp70 gene regulation and Hsp70 gene in the development of COPD.展开更多
Objective:To investigate the antifertility effect of chronically administered Tabernaemontana divarkala(T.divaricata) leaf extract on male rats.Methods:The effect of 50%ethanol extract of T.divaricata leaves on reprod...Objective:To investigate the antifertility effect of chronically administered Tabernaemontana divarkala(T.divaricata) leaf extract on male rats.Methods:The effect of 50%ethanol extract of T.divaricata leaves on reproduction was studied on male rats.The study was divided into four groups of five animals each.The first groups(1) received vehicle alone to serve as control. The second,third and fourth groups(Ⅱ,ⅡandⅣ) of animals were administered the leaf extract daily at 50 mg/kg body weight,p.o.,100 nig/kg body weight,p.o.,and 200 mg/kg body weight, p.o.,respectively,for a period of 60 days.Results:Significant decreases in the weight of testes, epididymis,seminal vesicle and ventral prostate were observed.A dose related reduction in the testicular sperm count,epididymal sperm count and motility,number of fertile male,ratio between delivered and inseminated females and numbers of pups were observed.The testis showed a clear correlation between the dose and severity of lesions of seminiferous epithelium. In general,the seminiferous tubules appear reduced in size with a frequently filled eosinophilic material.Spermatogenesis arrested at the secondary spermatocyte stage.Pachytene spermatocytes were undergoing degeneration.Disorganisation and sloughing of immature germ cell were visible. Leydinf cells were atrophied.No morphological changes were observed in Sertoli cells.Significant reduction in serum concentration of luteinizing hormone and testosterone were observed.No distinct change in serum FSH concentration was recorded.The final body weights of all groups were elevated markedly.No alterations were recorded in any hematologiocal parameters. Conclusions:It is concluded that the 50%ethanol extract of T.divaricata leaf produced dose related effect on male reproduction without altering general bodv metabolism.展开更多
Propolis is a resinous natural product,produced by bees(Apis mellifera),from vegetable parts and plant secretions.Propolis’samples A,B,C and D were extracted with phosphate buffer saline(PBS)or with 70%EtOH at pH val...Propolis is a resinous natural product,produced by bees(Apis mellifera),from vegetable parts and plant secretions.Propolis’samples A,B,C and D were extracted with phosphate buffer saline(PBS)or with 70%EtOH at pH values 8.0,7.2 and 6.4 followed by:(1)reverse-phase high-performance liquid chromatography(RP-HPLC)on Purospher®Star RP-18 column,the quantity of caffeic acid,chrysin,pinocembrin and galangin was determined;(2)determination of total flavonoids in both extracts;(3)antimicrobial tests of both extracts against(a)Gram-positive bacteria:methicillin-resistant Staphylococcus aureus(MRSA),St.aureus,Streptococcus pyogenes,Str.agalactiae,(b)Gram-negative bacteria:Escherichia coli,Pseudomonas aeruginosa,Proteus mirabilis,Acinetobacter baumanii and(c)yeast:Candida albicans.The antimicrobial activity of propolis’PBS extracts against Gram-positive bacteria shows the lowest minimal inhibitory concentration(MIC,mg/mL)at pH 8.0 in sample C,followed by A,B and D.In sample C,MICs at pH 8.0 were 0.007(Str.agalactiae),0.015(MRSA),0.015(Str.pyogenes)and 0.007(St.aureus).The polyphenol content of sample C is:flavonoid content 5.47±0.62 mg/mL,caffeic acid 1.33±0.92 mg/mL,chrysin 41.02±4.22μg/mL,pinocembrin 2.93±0.33 mg/mL and galangin 41.87±4.23 mg/mL.PBS extracts against Gram-negative bacteria show the lowest MIC(mg/mL)at pH 8.0 in sample D,followed by B,C and A.In sample D,MICs at pH 8.0 were 0.003(Acin.baumanii,Pr.mirabilis,Ps.aeruginosa)and 0.007(E.coli).Polyphenol content of sample D is:flavonoids 8.28±0.92 mg/mL,caffeic acid 3.56±0.32 mg/mL,chrysin 677.42±68.42μg/mL,pinocembrin 146.49±13.89 mg/mL and galangin 59.81±5.86 mg/mL.The strongest anti C.albicans activity,with the lowest MIC(mg/mL),at pH 8.0 was in the sample C,followed by samples D,A and B.In sample C,the MIC at pH 8.0 is 0.001(PBS extract).The antimicrobial activities of selected propolis samples correlate with their polyphenol content,more precisely,flavonoid,caffeic acid,chrysin,pinocembrin and galangin content.展开更多
Objective： To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction （当归补血汤, EDBD） was able to protect cardiomyocytes from oxidative injury of rats induced by hydro...Objective： To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction （当归补血汤, EDBD） was able to protect cardiomyocytes from oxidative injury of rats induced by hydrogen peroxide （H2O2） and its potential mechanism. Methods： Myocardial cells from 1- to 2-day-old neonatal rats were cultured in Dulbecco＇s modified Eagle＇s medium low-glucose and Ham＇s F12 medium （1：1）, and the cellular injury was induced by H2O2. The ultra-filtration extract mixture from Angelica sinensis and Hedysarurn po/ybotrys was given in three doses of 3.75, 7.5, and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （Ml-r） assay. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase （LDH） and creatine kinase （CK） releases in the medium, superoxide dismutase （SOD） activities, and intracellular malondialdehyde （MDA） and myeloperoxidase （MPO） contents. The levels of caspase-3 and heat shock protein 70 （hsp70） mRNA expression in cardiomyocytes were measured by reverse transcription polymerase chain reaction. Results： The EDBD could protect the cardiomyocytes from H202 injury in a dose- dependent manner （3.75, 7.50, and 15.00 mg/mL）. The EDBD could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents, increase SOD activity, up-regulate hsp70 expression, and down-regulate caspase-3 expression. Conclusion： The EDBD has protection on cardiomyocytes injured by H202 through improving cell antioxidant ability, up-regulating hsp70 expression, and inhibiting caspase-3 activity.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.8107003681370145+3 种基金81370156 and 81070021)The National Key Technology R&D Program of the 12th National Five-year Development Plan:Clinical Study on Translational Medicine of Respiratory Disease(No.212BA105B01)The Specific Project of National Health Research Project of Ministry of Health of China(No.201002008)Program for Changjiang Scholars and Innovative Research Team in University(No.PCSIRT1131)
文摘Cigarette smoke is associated with the development of several diseases, such as chronic ob- structive pulmonary disease （COPD）. The purpose of this study was to investigate genotoxicity and heat shock protein 70 （Hsp70） in human airway smooth muscle cells （HASMCs） exposed to cigarette smoke extract （CSE）. HASMCs was exposed to CSE with different doses for 24 h. The level of 8-hydroxydeoxyguanosine （8-OHdG） was determined by using HPLC-ECD, the DNA damage was ana- lyzed by using comet assay, and apoptosis was examined by using Annexin-FITC/PI staining. The pro- duction of Hsp70 after CSE stimulation was tested. Results indicated that CSE significantly increased the level of 8-OHdG, DNA damage and cell apoptosis, and reduced the production of Hsp70. In par- ticular, levels of Hsp70 were inversely correlated with 8-OHdG, DNA damage and cell apoptosis. It was concluded that cigarette smoke induced genotoxicity and decreased the production of cell protective protein Hsp70, which may contribute to the development of some airway diseases.
基金This project was supported by grants from the NationalKey Technologies R&D Program of the Tenth five year plan[2001BA703B03 ( B)], the Clinical Intensive Discipline of Ministry of Public Health ([ 2001 ] 321 ) and the National Natural Sciences Foundation of China (No. 30400194).
文摘The effects of cigarette smoke extract (CSE) on the expression of heat stress protein 70 (Hsp70) in human bronchi smooth muscle cells were investigated in vitro, and the changes in Hsp70 mRNA in the patients with chronic obstructive pulmonary disease and their significance were explored. Human bronchi smooth muscle cells were cultured with CSE at the different concentrations. The expression of Hsp70 mRNA and Hsp70 was detected by reverse translation-polymerase chain reaction (RT-PCR) and Western blotting respectively. Levels of Hsp70 mRNA and Hsp70 in lymphocytes from 20 patients with COPD and 20 healthy smoking control subjects were measured by RT-PCR and Western blotting. The results showed the expression of both Hsp70 mRNA and Hsp70 was decreased conformably in human bronchi smooth muscle cells treated with CSE at certain concentration in vitro. The A values of the Hsp70 mRNA expression were 0.24±0.11 and 0.42±0.13 respectively in COPD patients and healthy smoking controls with the difference being significant (P<0.01). There was also significant difference in the A values of the Hsp70 expression between COPD patients and healthy smoking controls (20.9±9.9 vs 44.8±15.3, P<0.01). The levels of Hsp70 mRNA had strongly positive correlation with Hsp70 protein (r = 0.85, P<0 01). It was suggested that the expression of Hsp70 mRNA was in concordance with the expression of Hsp70, which could provide a basis on the study of Hsp70 gene regulation and Hsp70 gene in the development of COPD.
文摘Objective:To investigate the antifertility effect of chronically administered Tabernaemontana divarkala(T.divaricata) leaf extract on male rats.Methods:The effect of 50%ethanol extract of T.divaricata leaves on reproduction was studied on male rats.The study was divided into four groups of five animals each.The first groups(1) received vehicle alone to serve as control. The second,third and fourth groups(Ⅱ,ⅡandⅣ) of animals were administered the leaf extract daily at 50 mg/kg body weight,p.o.,100 nig/kg body weight,p.o.,and 200 mg/kg body weight, p.o.,respectively,for a period of 60 days.Results:Significant decreases in the weight of testes, epididymis,seminal vesicle and ventral prostate were observed.A dose related reduction in the testicular sperm count,epididymal sperm count and motility,number of fertile male,ratio between delivered and inseminated females and numbers of pups were observed.The testis showed a clear correlation between the dose and severity of lesions of seminiferous epithelium. In general,the seminiferous tubules appear reduced in size with a frequently filled eosinophilic material.Spermatogenesis arrested at the secondary spermatocyte stage.Pachytene spermatocytes were undergoing degeneration.Disorganisation and sloughing of immature germ cell were visible. Leydinf cells were atrophied.No morphological changes were observed in Sertoli cells.Significant reduction in serum concentration of luteinizing hormone and testosterone were observed.No distinct change in serum FSH concentration was recorded.The final body weights of all groups were elevated markedly.No alterations were recorded in any hematologiocal parameters. Conclusions:It is concluded that the 50%ethanol extract of T.divaricata leaf produced dose related effect on male reproduction without altering general bodv metabolism.
基金the frame of Croatian Institute for Experimental and Translational Oncology(CIETO)supported by IvanČermak from the Crodux-plin,Savska Opatovina,10000 Zagreb,Croatia。
文摘Propolis is a resinous natural product,produced by bees(Apis mellifera),from vegetable parts and plant secretions.Propolis’samples A,B,C and D were extracted with phosphate buffer saline(PBS)or with 70%EtOH at pH values 8.0,7.2 and 6.4 followed by:(1)reverse-phase high-performance liquid chromatography(RP-HPLC)on Purospher®Star RP-18 column,the quantity of caffeic acid,chrysin,pinocembrin and galangin was determined;(2)determination of total flavonoids in both extracts;(3)antimicrobial tests of both extracts against(a)Gram-positive bacteria:methicillin-resistant Staphylococcus aureus(MRSA),St.aureus,Streptococcus pyogenes,Str.agalactiae,(b)Gram-negative bacteria:Escherichia coli,Pseudomonas aeruginosa,Proteus mirabilis,Acinetobacter baumanii and(c)yeast:Candida albicans.The antimicrobial activity of propolis’PBS extracts against Gram-positive bacteria shows the lowest minimal inhibitory concentration(MIC,mg/mL)at pH 8.0 in sample C,followed by A,B and D.In sample C,MICs at pH 8.0 were 0.007(Str.agalactiae),0.015(MRSA),0.015(Str.pyogenes)and 0.007(St.aureus).The polyphenol content of sample C is:flavonoid content 5.47±0.62 mg/mL,caffeic acid 1.33±0.92 mg/mL,chrysin 41.02±4.22μg/mL,pinocembrin 2.93±0.33 mg/mL and galangin 41.87±4.23 mg/mL.PBS extracts against Gram-negative bacteria show the lowest MIC(mg/mL)at pH 8.0 in sample D,followed by B,C and A.In sample D,MICs at pH 8.0 were 0.003(Acin.baumanii,Pr.mirabilis,Ps.aeruginosa)and 0.007(E.coli).Polyphenol content of sample D is:flavonoids 8.28±0.92 mg/mL,caffeic acid 3.56±0.32 mg/mL,chrysin 677.42±68.42μg/mL,pinocembrin 146.49±13.89 mg/mL and galangin 59.81±5.86 mg/mL.The strongest anti C.albicans activity,with the lowest MIC(mg/mL),at pH 8.0 was in the sample C,followed by samples D,A and B.In sample C,the MIC at pH 8.0 is 0.001(PBS extract).The antimicrobial activities of selected propolis samples correlate with their polyphenol content,more precisely,flavonoid,caffeic acid,chrysin,pinocembrin and galangin content.
基金Supported by the Key Projects of the National Science and Technology Pillar Program during the Eleventh Five-Year Plan Period(No.2007BAI37B01)Major Science and Technology Program of Gansu Province(No.092NKDA017)
文摘Objective： To investigate whether the administration of the ultra-filtration extract from Danggui Buxue Decoction （当归补血汤, EDBD） was able to protect cardiomyocytes from oxidative injury of rats induced by hydrogen peroxide （H2O2） and its potential mechanism. Methods： Myocardial cells from 1- to 2-day-old neonatal rats were cultured in Dulbecco＇s modified Eagle＇s medium low-glucose and Ham＇s F12 medium （1：1）, and the cellular injury was induced by H2O2. The ultra-filtration extract mixture from Angelica sinensis and Hedysarurn po/ybotrys was given in three doses of 3.75, 7.5, and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （Ml-r） assay. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase （LDH） and creatine kinase （CK） releases in the medium, superoxide dismutase （SOD） activities, and intracellular malondialdehyde （MDA） and myeloperoxidase （MPO） contents. The levels of caspase-3 and heat shock protein 70 （hsp70） mRNA expression in cardiomyocytes were measured by reverse transcription polymerase chain reaction. Results： The EDBD could protect the cardiomyocytes from H202 injury in a dose- dependent manner （3.75, 7.50, and 15.00 mg/mL）. The EDBD could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents, increase SOD activity, up-regulate hsp70 expression, and down-regulate caspase-3 expression. Conclusion： The EDBD has protection on cardiomyocytes injured by H202 through improving cell antioxidant ability, up-regulating hsp70 expression, and inhibiting caspase-3 activity.
