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OBJECTIVE@#To investigate the effect of inhibitor of growth protein-2 (Ing2) silencing on angiotensin Ⅱ (AngⅡ)-induced cardiac remodeling in mice and explore the underlying mechanism.@*METHODS@#An adenoviral vector carrying Ing2 shRNA or empty adenoviral vector was injected into the tail vein of mice, followed 48 h later by infusion of 1000 ng · kg-1 · min-1 Ang Ⅱ or saline using a mini-osmotic pump for 42 consecutive days. Transthoracic echocardiography was used to assess cardiac geometry and function and the level of cardiac hypertrophy in the mice. Masson and WGA staining were used to detect myocardial fibrosis and cross-sectional area of cardiomyocytes, and myocardial cell apoptosis was detected with TUNEL assay. Western blotting was performed to detect myocardial expressions of cleaved caspase 3, ING2, collagen Ⅰ, Ac-p53(Lys382) and p-p53 (Ser15); Ing2 mRNA expression was detected using real-time PCR. Mitochondrial biogenesis, as measured by mitochondrial ROS content, ATP content, citrate synthase activity and calcium storage, was determined using commercial assay kits.@*RESULTS@#The expression levels of Ing2 mRNA and protein were significantly higher in the mice with chronic Ang Ⅱ infusion than in saline-infused mice. Chronic infusion of AngⅡ significantly increased the left ventricular end-systolic diameter (LVESD) and left ventricular end-diastolic diameter (LVEDD) and reduced left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) in the mice. Ing2 silencing obviously alleviated AngⅡ-induced cardiac function decline, as shown by decreased LVEDD and LVESD and increased LVEF and LVFS, improved myocardial mitochondrial damage and myocardial hypertrophy and fibrosis, and inhibited cardiomyocyte apoptosis. Chronic AngⅡ infusion significantly increased myocardial expression levels of Ac-p53(Lys382) and p-p53(Ser15) in the mice, and Ing2 silencing prior to AngⅡ infusion lessened AngⅡ- induced increase of Ac-p53(Lys382) without affecting p53 (ser15) expression.@*CONCLUSION@#Ing2 silencing can inhibit AngⅡ-induced cardiac remodeling and dysfunction in mice by reducing p53 acetylation.
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Animals , Mice , Angiotensin II , Tumor Suppressor Protein p53 , Acetylation , Stroke Volume , Ventricular Remodeling , Ventricular Function, Left , Myocytes, CardiacABSTRACT
Objective@#To study the influence of meteorological factors on the incidence of hand foot and mouth disease (HFMD) in Xiamen, Fujian province, and provide scientific evidence for the early warning, prediction, prevention and control of HFMD.@*Methods@#Correlation analysis and distribution lag nonlinear models (DLNM) analysis of meteorological factors such as daily average pressure, daily average relative humidity, daily average temperature and sunshine hours and the incidence of HFMD in Xiamen during 2013 to 2017 were conducted by using R3.4.3 software.@*Results@#A total of 36 464 cases of HFMD were reported in Xiamen during 2013-2017, and the incidence showed an upward trend (F=40.359, P=0.008). The daily average relative humidity, daily average temperature and sunshine hours were positively correlated with the incidence of HFMD (r>0), and the daily average site pressure was negatively correlated with the incidence of HFMD (r<0). In the case of a lag of 0-5 days, when the daily average pressure of the station was higher than 1 005 hPa, the risk of HFMD gradually increased with the increase of air pressure, and the risk of disease decreased with the increase of lag days. The risk was highest when air pressure was 1 017 hPa and at the lag of 0 day (RR=1.14, 95%CI: 0.67-1.94). When the relative humidity was higher than 95%, the risk of HFMD gradually increased with the increase of relative humidity, and the lag time ranged from 0 day to 10 days, which was most obvious on the 4th and 5th days. The risk was highest when relative humidity was 100% and at the lag of 5 days (RR=1.32, 95%CI: 1.02-1.71). When the air temperature was >28 ℃ and <8 ℃, the risk of HFMD existed, but the lag time was inconsistent. The relative risk was highest during 15-20 days at low air temperature, and the lag time at high air temperature was mainly during 5-15 days. The risk was highest when air temperature was 28 ℃ and at the lag of 4 days (RR=1.10, 95%CI: 0.94-1.29). The sunshine time was >12 h and lag of 0-3 days was a risk factor for the incidence of HFMD. The risk was highest when sunshine time was 13 h and the lag of 0 day (RR=1.20, 95%CI: 1.05-1.36).@*Conclusion@#Meteorological factors such as daily average pressure, daily average relative humidity, daily average temperature and sunshine hours were associated with the incidence of HFMD with certain lag in Xiamen. So, it is suggested to use these data in the early warning system of HFMD.
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Objective To investigate the mechanism of epidermal growth factor receptor-forkhead transcription factor A2 (EGFR-FOXA2) pathway-involved high secretion of mucus in human bronchial epitheli-um (HBE) cells after respiratory syncytial virus (RSV) infection and to evaluate the effects of intervention using agonist ( rosiglitazone ) and antagonist ( GW9662 ) of peroxidase proliferation activated receptor γ( PPARγ) and EGFR inhibitor ( AG1478 ) . Methods HBE cells were randomly divided into six groups: A group ( AG1478+RSV) , B group ( rosiglitazone+RSV) , C group ( GW9662+RSV) , D group ( RSV) , E group (0. 1% dimethyl sulfoxide DMSO) and F group (HBE cell control group). Two hours before RSV infection, A, B and C groups were respectively treated with 10 μmol/L of AG1478, rosiglitazone and GW9662. Expression of EGFR, PPARγ and FOXA2 at mRNA level in each group was detected by real-time fluorescence quantitative RT-PCR 12 h, 24 h and 48 h after HBE cells were infected with or without RSV. Expression of phosphorylated-EGFR ( p-EGFR) and EGFR at protein level was detected by Western blot. ELISA was performed to measure the expression of mucin-5AC (MUC5AC). Results Compared with F group, EGFR expression at mRNA lev-el, p-EGFR/EGFR protein ratio and MUC5AC expression at protein level were increased in a time-dependent manner in A, B, C and D groups at 12 h, 24 h and 48 h. Compared with group F, the expression of PPARγat mRNA level in A, B, and D groups increased at each time point. Moreover, PPARγ expression gradually in-creased over time in A and B groups, reaching the peaks at 48 h, but was in decline in D group. Expression of FOXA2 at mRNA level in RSV-infected HBE cells was declined at each time point compared with that in group F, especially in D group. Compared with group D, A and B groups showed significantly decreased EGFR ex-pression at mRNA level, p-EGFR/EGFR protein ratio and MUC5AC expression at protein level, but markedly increased FOXA2 expression at mRNA level. Conclusions RSV infection increased the expression of MUC5AC at protein level in HBE cells. PPARγand EGFR-FOXA2 signaling pathways were involved in the hypersecretion of airway mucus during RSV infection.
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OBJECTIVE To investigate the effect of asiaticoside (AS) on endothelial-to-mesenchymal transition (EndoMT) in hypoxia pulmonary hypertension (HPH). METHODS Male Sprague-Dawley (SD) rats were divided into normoxia control group, hypoxia model group, and AS 25 and 50 mg · kg-1 group. Hypoxia model group and AS group were subjected to intermittent hypoxia exposure. Control group and model group received 1-1.5 mL saline daily, and AS groups were ig administrated with AS 25 and 50 mg·kg-1 for 4 weeks. Human pulmonary artery endothelial cells (HPAECs) were divided into normoxia control group and hypoxia AS groups. Hypoxia groups were cultured with AS 0, 25, 50, 100 and 200 mg·L-1 for 72 h under hypoxic (5%O2, 5%CO2) conditions. Anti-proliferation effect of AS was investigated by CCK-8 assay. Then, HPAECs were divided into normoxia control group, normoxia AS 100 mg · L-1 group, hypoxia model group, and hypoxia AS 100 mg · L-1 group. After five days of culture, migration ability of cells was detected by Transwell test. Expression of CD31 andα-SMA was detected by immunofluorescence and Western blotting in both in vivo and in vitro experiments. RESULTS In both in vivo and in vitro experiments, compared with normoxia control group, expression of CD31 was reduced (P<0.01) andα-smooth muscle actin (α-SMA) was increased (P<0.01) in hypoxia model group in both immunofluorescent analysis and Western blotting. Compared with hypoxia model group, expression of CD31 was increased andα-SMA was decreased (P<0.05, P<0.01) in AS treatment groups. Compared with normoxia control group, proliferation and migration ability of HPAEC were elevated in hypoxia model group (P<0.05). Compared with hypoxia group, AS 100 mg · L-1 depressed proliferation and migration of HPAEC under hypoxia exposure up to 72 h (P<0.05). CONCLUSION EndoMT might be involved in HPH and could be partly inhibited by AS.
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OBJECTIVE To study the therapeutic effect and the underlying mechanism of asiatico?side on bleomycin-induced rat interstitial pulmonary fibrosis(IPF). METHODS Male Sprague-Dawley (SD)rats were divided into normal control group,bleomycin 5 mg·kg-1 model group and asiaticoside 50 mg · kg-1 group. The model and asiaticoside group were administrated with bleomycin 5 mg · kg-1 to induce IPF,while the asiaticoside group was administrated with asiaticoside 50 mg·kg-1 by gastric perfusion. Hematein eosin(HE)and Masson staining were carried out to analyze the histopathological changes in the lung. Lung homogenates were used to examine hydroxyproline(HYP) content,and serum samples were used to measure the concentration of interferon-γ(IFN-γ),interleukin-4(IL-4) and tumor necrosis factor-α(TNF-α). In addition,immunohistochemical methods were used to locate lung transforming growth factor-β1(TGF-β1)and adenosine 2A receptor(A2AR)expression,and Western blotting was used to examine the expression levels of TGF-β1 and A2AR. RESULTS On the 7th,14th and 28th days,the scores of pulmonary inflammation were higher in model group than in control group (P<0.01),and the asiaticoside group showed mitigated alveolitis(P<0.01,P<0.05) compared with model group. Compared with control group,the scores of pulmonary fibrosis in model group were elevated(P<0.01),and the asiaticoside group showed reduced pulmonary fibrosis(P<0.05). On the 14th and 28th days,HYP content in the model group〔1.85±0.10,(2.48±0.18)mg·g-1〕was higher than in the control group〔0.79 ± 0.07,(0.84 ± 0.08)mg · g-1〕(P<0.01),but HYP content in the asiaticoside group〔1.32±0.131,(1.71±0.13)mg·g-1〕was lower than in the model group(P<0.05). IL-4 and TNF-αin the asiaticoside group were lower than in model group(P<0.05),but were higher in the model group than in the control group(P<0.01,P<0.05). The expression level of TGF-β1 protein in the asiaticoside group was lower than in the model group(P<0.05),but was higher in the model group than in the control group(P<0.05). The expression level of A2AR protein in the asiaticoside group was higher than in the model group(P<0.05),but was lower in the model group than in the control group(P<0.05). CONCLUSION Asiaticoside can mitigate bleomycin-induced IPF by inhibiting the expression of IL-4, TNF-αand TGF-β1,and raising the level of A2AR.
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Objective To investigate clinical effects of early postoperative oral enteral nutrition on immune function for colorectal cancer patients,to provide a reference for clinical treatment.Methods We selected 90 colorec-tal cancer surgery patients from January 2012 to April 2015,and they were divided into two groups by random number method.50 cases in the study group were given early oral enteral nutrition,and 40 patients in the control group were given postoperative parenteral nutrition.Before and after treatment,the quality of life,complications,immunology indi-cators,nutrition indicators were compared in two groups.Results In the observation group,the postoperative fever, exhaust time,hospital stay,hospital costs,quality of life scores were (54.29 ±5.76)h,(57.89 ±5.95)h,(10.48 ± 1.63)d,(41 432.85 ±2 165.46)RMB,(79.23 ±5.17)points,which were significantly lower than those in the con-trol group,the differences were statistically significant (t =8.69,P =0.00;t =8.61,P =0.00;t =9.49,P =0.00;t =5.54,P =0.00;t =4.16,P =0.01);7d after the treatment,the IgA,IgG,IgM,CD +4 ,CD +4 /CD +8 in observation group were (1.92 ±0.49)g/L,(11.31 ±2.24)g/L,(0.99 ±0.21)g/L,(41.21 ±5.51)%,(1.72 ±0.32),which were better than those in the control group,the differences were statistically significant (t =2.05,P =0.04;t =2.11, P =0.04;t =2.12,P =0.04;t =3.64,P =0.01;t =2.23,P =0.03).In observation group,postoperative 7d Hb, TRF,ALB,PAB were (113.28 ±13.36)g/L,(3.02 ±0.39)mg/L,(38.97 ±3.15)g/L,(333.15 ±18.35)mg/L, which were better than those in the control group,the differences were statistically significant (t =3.45,P =0.01;t =2.18,P =0.03;t =2.32,P =0.04;t =3.21,P =0.01).7d after operation,the CRP level of observation group was(7.29 ±2.05)mg/L,which was significantly lower than that of the control group,the difference was statistically sig-nificant (t =4.22,P =0.01 ).The incidence rate of postoperative complications in the observation group was 26.00%,that of the control group was 40.00%,the difference was statistically significant (χ2 =4.52,P =0.01). Conclusion Early oral enteral nutrition in patients with colorectal cancer will help to improve immune function, nutritional support,reduce the inflammatory response,which will help postoperative recovery.
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<p><b>OBJECTIVE</b>A Dengue outbreak was reported in Dongfen town Jianou county, Fujian province on September 19, 2014. The goal of this project was to explore the role of syndromic surveillance program in the practice of early detection on disease outbreak through the case mentioned above.</p><p><b>METHODS</b>The authors retrospectively collected data related to Outpatient log and Pharmacy drug use in Dongfen township hospital through the electronic information system of the hospital from August to November, 2014. All the abnormal events were recorded, according to related data on fever and drug use. Description of fever, syndromic characteristics, correlation and Linear regression analyses were conducted, using the surveillance data on fever syndrome and drug use from the pharmacy.</p><p><b>RESULTS</b>A total of 1 102 cases with fever and 2 437 fever-related clinic visits were reported which showing an increased number of 19.6, 10.2 times respectively, when compared to the same period of the previous year in which men accounted for 45.3% (499/1 102) and female accounted for 54.7% (603/1 102). Age groups presented an atypical type " M" type. 5 and 10 year olds groups formed the largest proportion, accounted for 11.5% (127/1 102) of the total number os the patients. The correlation coefficient ranged from 0.85 to 0.97 (P<0.05). Data from the syndromic surveillance program showed an " outbreak" was occured in August 23, 2014.</p><p><b>CONCLUSIONS</b>Compared to routine surveillance program, the syndromic surveillance program could detect the appearence of an outbreak, a month or even more earlier. The role of syndromic surveillance program needs to be further explored.</p>
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Female , Humans , Male , Data Collection , Dengue , Diagnosis , Epidemiology , Disease Outbreaks , Drug Prescriptions , Drug Utilization , Early Diagnosis , Fever , Health Information Systems , Pharmacy Service, Hospital , Population Surveillance , Methods , Retrospective StudiesABSTRACT
The Middle East Respiratory Syndrome (MERS) discovered in 2012 was caused by a novel coronavirus .With high mortality and severe acute respiratory syndrome ,the coronavirus had been named as the Middle East Respiratory Syn‐drome Coronavirus (MERS‐COV) .At present ,the disease occurred in the Middle East and spread to other countries aroused extensive attention all over the world .In this review ,we mainly focus on the research progress of MERS‐CoV about the epide‐miological characteristics ,clinical treatment and global response situation ,providing advices and references of preventing and controlling effectively for M ERS in the future .
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Objective: To investigate the effect of the gene interfering technology on fatty acid synthase (FAS) gene silencing for lipid contents in human hepatic cell line HepG2 and to study the lipid metabolism related gene expression in HepG2 cells. Methods: A total of 3 pairs of small interfering RNA (siRNA) targeting different sequences of FAS mRNA were synthesized as FAS-siRNA-1, FAS-siRNA-2 and FAS-siRNA-3, meanwhile, 2 controls were established as Blank control group, in which HepG2 cells were not treated, and Negative control group, in which HepG2 cells were transfected by non-effective siRNA. The mRNA, and protein expression levels of FAS in HepG2 cells were examined by real-time lfuorescence quantitative RCR and Western blot analysis to screen the most effective pair of siRNA for FAS gene silencing; and that speciifc siRNA was transtected to HepG2 cells for 48 hours to detect the intra-/extra-cellular TG, TC levels and the mRNA expression related to lipid metabolism in HepG2 cells. Results: The screening experiment indicated that FAS-siRNA-3 was most effective for FAS gene silencing. Compared with Blank control group, the mRNA and protein expressions in FAS-siRNA-3 transfected HepG2 cells (Transfected group)decreased to (52.33 ± 3.07) % and (51.57 ± 3.14) % respectively. Compared with Blank control group, Transfected group had the reduced intra-/extra-cellular TG levels and reduced extracellular TC level; while increased mRNA expression of hepatic lipase,P<0.0001 and decreased mRNA expression of TG transfer protein in HepG2 microsome,P<0.05. Conclusion: FAS gene silencing could signiifcantly decrease the intra-/extra- cellular TG level and extracellular TC level in HepG2 cells, those ifndings need to be conifrmed by furtherin vivo andin vitro studies.
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Objective To detect the prevalence of human cytomegalovirus (HCMV) in peripheral blood of patients with systemic lupus erthematosus (SLE) and explore its role in the pathogenesis of SLE.Methods HCMV DNA was isolated from the peripheral blood leucocytes (PBLs) of 60 patients with SLE and 111 healthy controls.Nested polymerase chain reaction (nPCR) technology was used to investigate the gene of HCMV glycoprotein gB (UL55) in these specimens.HCMV infections in the PBLs of SLE patients were confirmed by HCMV-UL55 detection.Two-sample t test,nonparametric test,Chi-square test and Fisher probabilities were used to analyze.Results Agarose gel electrophoresis and sequencing analysis showed that established nPCR could specifically detect HCMV-UL55 gene,the HCMV infection rate was significantly higher in patients with SLE than in the healthy controls (P<0.01).Positive rates of HCMV infection in SLE group and controls were 41.7% (25/60) and 1.8% (2/111),respectively.Compared to the SLE patients with HCMV-negative PBLs,the positive rate of Rib-P [26%(9/35) vs 56%(14/25),x2=5.659,P=0.017],the positive rate of direct Coomb's test [37%(13/35) vs 72%(18/25),x2=7.096,P=0.008] and the level of antiβ2GP1 [21.3 (9.9,51.8) U/ml vs 13.6 (5.9,23.1) U/ml,U=2.017,P=0.044] were significantly higher than those in the SLE patients with HCMV-positive PBLs.Compared to the SLE patients with HCMV-negative PBLs,the number of red blood cells [(3.65±0.10)×1012/L vs (3.17±0.17)×1012/L,t=2.574,P=0.013] and lymphocytes [(1.37±0.14)×1012/L vs (0.90±0.13)×1012/L,t=2.456,P=0.017] in peripheral blood and the hemoglobin levels [(110±19) g/L vs (98±5)g/L,t=2.034,P=0.048] of the SLE patients with HCMV-positive decreased significantly.At the same time,the positive rate of hematuria [40%(14/35) vs 72%(18/25),x2=6.000,P=0.014] and 24 h proteinuria [0.80 (0.53,2.37)g vs 0.48 (0.13,1.21)g,U=2.140,P=0.032],which indicated kidney damage were also significantly increased in SLE patients with HCMV-positive PBLs.Conclusion The infection of HCMV in peripheral blood cells may take part in the development of SLE.
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<p><b>OBJECTIVE</b>To explore the prevalence and the different risk factors for asthma in children between urban and rural areas in Fuzhou, Fujian province.</p><p><b>METHOD</b>The epidemiological survey of asthma in 0-14 years old children was conducted from October 2009 to October 2010 between Fuzhou urban and rural areas in Fujian province. The investigation subjects were selected in urban and rural areas by phased stratified random cluster sampling. The 2010 third national epidemiological survey questionnaire of children with asthma was used for screening for possible patients. Diagnosis of asthma was confirmed by physical examination. The children with asthma were designated as the positive cases, while non asthmatic children who were age, gender, ethnic, and living environment matched with asthmatic patients were designated as negative control. Comparison of the prevalence of asthma in children between Fuzhou urban and rural areas was performed. The influencing factors of asthma were analyzed and screened by the regression equation model of two element Logistic regression.</p><p><b>RESULT</b>Totally 12 235 questionnaires of children with asthma and allergic disease screening were issued and 11 738 questionnaire were sent back (6 221 were male and 5 517 were female). The return rate was 95.9% in urban Fuzhou; 648 children were diagnosed as asthma. The prevalence of asthma in male was 6.48% and female children was 4.44% (comparison of the prevalence of gender χ(2) = 23.267, P < 0.001) in urban areas . A total of 6 000 questionnaires of children with asthma and allergic disease screening were sent out and 5 860 were responded (male children 3 228, female children 2 632). The recovery rate was 97.7% in rural Fuzhou; 135 children with asthma was diagnosed. The prevalence of asthma in male was 2.73%and female children and was 1.79%. Adding protein supplement before 6 months (OR = 1.908, 95%CI:1.233-2.959), the use of antibiotics in the treatment of asthma (OR = 14.541, 95%CI:8.920-23.705), furniture materials (non wood) (OR = 2.432, 95%CI:1.563-3.785) were the main risk factors of children with asthma in urban. Adding protein supplement before 6 months(OR = 3.021, 95%CI:1.357-6.711), the use of antibiotics in the treatment of asthma(OR = 14.784, 95%CI:3.842-56.885), the use of coal as fuel (OR = 63.339, 95% CI: 7.993-501.943), domesticated livestock (OR = 13.659, 95% CI:1.342-139.068), the family smoking before and after birth (OR = 6.226, 95%CI:2.674-14.495) and chemical fiber pillow (OR = 3.638, 95%CI:1.241-10.666) were the main risk factors of children with asthma in rural areas.</p><p><b>CONCLUSION</b>The prevalence of children with asthma in urban areas was higher than that in rural areas. The prevalence of asthma in male children was higher than in female children. Adding protein food supplement before 6 months, the use of antibiotics and non solid wood furniture material were the main risk factors in children with asthma in urban areas. Adding protein supplement before 6 months, the use of antibiotics, domesticated livestock, the use of coal as fuel and the family smoking before and after birth were the main risk factor of asthma in children in rural areas.</p>
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Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Anti-Bacterial Agents , Asthma , Epidemiology , China , Epidemiology , Logistic Models , Prevalence , Risk Factors , Rural Population , Sampling Studies , Smoking , Surveys and Questionnaires , Urban PopulationABSTRACT
To observe the influence of human leucocyte antigen B27 [HLA-B27] status and gender on sacroiliitis on computed tomography [CT] in ankylosingspondylitis [AS]. We reviewed the archived medical records of the AS inpatients admitted in the Rheumatology Department of the First Affiliated Hospital of Wenzhou Medical University during the period from January 2007 through January 2013 and finally 386 patients were included in the study. The severity of sacroiliitis on CT was evaluated according to the grading used in the modified New York criteria for AS. Two-way classification analysis of variance [ANOVA] was employed to examine the effect of HLA-B27 status and gender on age at disease onset. The impact of HLA-B27 and gender on sacroiliitis on CT was tested by univariate and multivariate logistic regression analyses. There were 350 HLA-B27 positive patients [90.7%] and 36 HLA-B27 negative patients [9.3%]. The ANOVA test indicated that HLA-B27 positive patients and male patients respectively had an earlier age at disease onset than HLA-B27 negative patients and female patients. The logistic regression analysis indicated that positive HLA-B27 status [OR 2.601, p=0.004] and male gender [OR 1.923, p=0.004] were significant predictors of worse sacroiliitis. In addition, elevated ESR [OR 2.181, p=0.013] and longer disease duration [OR 1.100, p < 0.001] contributed to worse sacroiliitis likewise. Positive HLA-B27 status and male gender are associated with worse sacroiliitis on CT, acting as predictors of sacroiliitis. Elevated ESR and longer disease duration also contribute to worse sacroiliitis. Meanwhile, positive HLA-B27 status and male gender are associated with earlier age at disease onset
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Objective To observe the effect of hydrogen on ultraviolet B (UVB)-induced oxidative damage to skin fibroblasts.Methods Primary human skin fibroblasts from foreskin tissues were divided into five groups:normal control group receiving no treatment,hydrogen control group treated with hydrogen-rich saline,UVB group receiving irradiation only,post-treatment group irradiated with UVB followed by hydrogen-rich saline treatment,and pre-treatment group treated with hydrogen-rich saline followed by UVB irradiation.The dose of UVB was 30,60 and 90 mJ/cm2 in the cell proliferation assay and 90 mJ/cm2 in the other experiments.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of fibroblasts,a chemiluminescence method to estimate the activity of superoxide dismutase (SOD) and catalase as well as to determine the level of malondialdehyde in the culture supernatant of fibroblasts,enzyme linked immunosorbent assay (ELISA) to determine the supernatant level of 8-isoprostane-prostaglandin F2α (8-iso-PGF2α),Western blot to detect the expression of heme oxygenase-1 (HO-1) in fibroblasts.One-factor analysis of variance was conducted to assess differences in these parameters among these groups.Results UVB irradiation decreased the proliferative activity (absorbence value at 490 nm) of fibroblasts in a dose-dependent manner.Both the pre-treatment group and post-treatment group showed a statistical increase in proliferative activity of cells compared with the corresponding UVB control groups (all P < 0.05).The activity of SOD and catalase as well as the protein expression of HO-1 were significantly higher (all P < 0.05),whereas the supernatant levels of malondialdehyde and 8-iso-PGF2α were statistically lower (both P < 0.05) in the pre-treatment group and post-treatment group than in the UVB control group.Conclusion Hydrogen may mitigate UVB-induced oxidative damage to skin fibroblasts.
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Objective To investigate the proper inner control genes suitable for mRNA expression level comparison of aging rat tissues.Methods Real-time reverse transcription PCR was used to examine in aging rat tissues the expression level of G3pd(glyceraldehyde-3-phosphate dehydrogenase),ACTB(?-actin),H3f3b(H3 histone,family 3B),Arbp(acidic ribosomal phosphoprotein P0)and 18S(18S ribosomal RNA).Results The most stably expressed housekeeping gene in aging rat kidney was ACTB,in heart and lung G3pd showed the minimum variation;Arbp expression was the most stable one in different tissues.Conclusion For aging rat intra-tissue mRNA normalization at least two housekeeping genes should be used: one is the ribosomal RNA gene 18S and another one is Arbp.
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Objective To find an ideal method of DNA isolation from blood and especially from clotted blood and to minimize the volume of blood collected for laboratory and clinical tests.Methods DNAs were isolated from antiagglutinated and agglutinated blood samples from auricular veins of 30 healthy subjects. The DNAs of these samples were obtained by a nonenzymatic, nontoxic procedure optimized by us and determinated by agarose gel electrophoesis and PCR. Results The yields of DNA isolated from clotted blood and antiagglutinated blood were (40.2?8.86)mg DNA/L and (39.1?10.2)mg DNA/L, and purities were 1.87?0.11 and 1.92? 0.12. The DNAs that we isolated from all samples had high molecular weight and by PCR the dimorphism of ALU alleles of the 8th intron of t-PA was easy to be obtained, so they were complete and reliable. Conclusion This method is rapid, easy, efficient and nontoxic for isolation of DNA from clotted and fresh blood and meets requirements for clinical testing and molecular biology study.
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To keep vocational schools distinctive and vigorous,top priority should be placed in teachers'training and teaching,making practice skill training systematic,embodying practical open professional principle based on vocational education in the process of practice teaching,by the teachers'guidance and stimulating students to actively participate in practice,reforming teaching ways and means.
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AIM: To investigate the changes of bcl-2, bax expression and neuron apoptosis of cerebral cortex in lymphostatic encephalopathy of rats. METHODS: The model of lymphostatic encephalopathy was established by occluding and removing both the shallow and deep cervical lymph nodes in rats. The animals were sacrificed at 1, 2, 3, 5, 7 and 14 days after operation. HE staining was used to observe the structure of brain tissues and TUNEL staining was used to detect in situ cell apoptosis. The expressions of bcl-2 and bax were examined by RT-PCR. RESULTS: Cerebroedema appeared at the second day and was the most serious at the 5th day after blockage of cervical lymphatics. The number of TUNEL positive cells and the expression of bax began to increase at the 2nd day, reached a peak at the 5th day and dropped to control level at the 14th day. The expression of bcl-2 began to increase at the 1st day, reached a peak at the 5th day and dropped to control level at the 7th day. The increasing extent of bax was higher than that of bcl-2. CONCLUSION: The blockage of cervical lymphatics can lead to lymp[JP2]hostatic encephalopathy. Apoptosis is the main form of neuron death in the cortex and has relation to the increasing expression of bcl-2 and bax. [JP]