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Objective: To understand the relationship between female breast cancer and occupational risk factors in Beijing, and provide scientific basis for the formulation of prevention strategies and measures. Methods: From June to December 2019, A 1: 1 case-control study was adopted, eight medical institutions in Beijing were selected as the research objects. Patients with breast cancer diagnosed by medical institutions were selected as case group and non breast cancer patients in the same medical institution as control group. A total of 973 subjects were included, including 495 in the case group and 478 in the control group. A one-to-one survey was conducted using a questionnaire uniformly compiled by the Beijing Centers for Disease Control and Prevention. The survey content mainly includes basic demographic characteristics and occupational risk factors. The Trait Coping Style Questionnaire (TCSQ) was used to investigate the corresponding methods, including two aspects: positive coping and negative coping. First, chi square test or Wilcoxon rank sum test were used for univariate analysis. Then Lasso regression was used to screen the risk factors of breast cancer. Finally, the risk factors were screened by multivariate logistic regression analysis. Results: Education lovel was 49.64%, body mass index (BMI) was 18.4~23.9 kg/m(2), accounting for 48.82%, marital status ws 84.48%. Compared with no night shift history, there was a significant increase in risk of breast cancer at night shift history (OR=1.70, 95% CI: 1.25~2.30, P<0.05) . Compared with most of the sitting posture and sometimes standing, the risk of breast cancer was increased, and the difference was statistically significant (OR=2.01, 95%CI: 1.40~2.90, P<0.05) . Conclusion: In the occupation risk factors, night shift work and working posture are related to the incidence of breast cancer in women, establishing a good schedule and avoiding long standing can effectively prevent and reduce the occurrence of breast cancer.
Subject(s)
Female , Humans , Beijing , Breast Neoplasms/epidemiology , Case-Control Studies , Logistic Models , Risk FactorsABSTRACT
OBJECTIVE: To analyze the active surveillance results of occupational pneumoconiosis(hereinafter referred to as “pneumoconiosis”) in Beijing in 2019. METHODS: A total of 2 634 dust exposed workers were recruited as the active surveillance subjects by judgment sampling method. The abnormalities and influencing factors of High kV or digital photography of posterior-anterior chest radiography(hereinafter referred to as “chest radiography”) and lung function were analyzed. RESULTS: The detection rate of abnormal chest radiograph and pulmonary dysfunction were 14.0%(368/2 634) and 6.6%(175/2 634), respectively. The multivariate logistic regression analysis showed that the risk of abnormal chest radiograph in dust-exposed workers increased with age(P<0.01). Mining and construction workers had higher risk of abnormal chest radiograph than manufacturers(all P<0.05). The risk of abnormal chest radiograph of dusk-exposed workers in state-owned and foreign enterprises was higher than that of workers in private enterprises(P<0.05). The risk of pulmonary dysfunction was increased with age and length of dust exposure(all P<0.05). Workers exposed to silica dust and aluminum dust had higher risks than those exposed to welding dust(all P<0.01). The risk of pulmonary dysfunction of workers in small and micro enterprises was higher than that of workers in large enterprises(all P<0.01). Mining workers had higher risks of pulmonary dysfunction than manufacturing workers(P<0.05). CONCLUSION: Age, length of dust exposure, dust type, industry type, enterprise scale and economic type were the influencing factors of lung injury of dust exposed workers. Therefore, the supervision of key population and industries should be strengthened to reduce the occurrence of pneumoconiosis.
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ObjectiveTo analyze the endemic situation of schistosomiasis in national surveillance sites of Zhenjiang City from 2015 to 2019, so as to provide the scientific basis for the consolidation of schistosomiasis control achievements and timely treatment of risk factors. MethodsA total of 7 national schistosomiasis surveillance sites were assigned in Zhenjiang City from 2015 to 2019. According to the National Scheme for Schistosomiasis Surveillance (2014 version), Schistosoma japonicum infections were monitored in local residents, floating populations and livestock, and snail habitats were surveyed in Zhenjiang City from 2015 to 2019. The results of schistosomaisis surveillance in Zhenjiang City from 2015 to 2019 were analyzed. Results There were 15 848 and 7 779 person-times of local residents and floating populations receiving serological screening of schistosomiasis in 7 national surveillance sites of Zhenjiang City from 2015 to 2019, and the seroprevalence rates were 0.95% to 1.75% and 0.14% to 0.91% among local residents and floating populations, respectively; however, no egg-positives were identified. A total of 1 885 herd-times of livestock were detected, and no egg-positives were found. During the 5-year period, snail survey was conducted in snail habitats and suspected snail habitats covering 3 333.79 hm2, and there were 108.71 hm2 snail habitats identified, including 50.97 hm2 re-emerging snail habitats. The density of living snails was 0.007 to 0.125 snails/0.1 m2 from 2015 to 2019, which showed a trend for a rise year by year (χ2 = 2 732.626, P < 0.01); however, no S. japonicum infections were detected in snails. Conclusions The endemic situation of schistosomiasis has been maintained at a low level in Zhenjiang City, and schistosomiasis control achievements are consolidated. However, snails are widely distributed in the city, and there are still risk factors associated with schistosomiasis transmission. Further integrated interventions require to be reinforced to timely manage the risk of transmission of schistosomiasis and promote the progress towards elimination of schistosomiasis.
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Objective To understand the epidemic trend of soil-transmitted nematodiasis in Zhenjiang City from 2006 to 2018, so as to provide evidence for formulating targeted control measures. Methods The permanent residents at ages of 3 years and greater and the immigrants who had lived in Zhenjiang City for more than one year were selected as the study subjects. Fecal samples were collected, and the soil-transmitted nematode eggs were detected in fecal samples with the modified Kato-Katz technique (three smears from one stool sample), while the children at ages of 12 years and lower were tested for Enterobius vermicularis eggs using the adhesive cellophane-tape perianal swab method. The prevalence and intensity of parasitic infections were estimated in Zhenjiang City each year from 2006 to 2018, and the data pertaining to the deworming for soil-transmitted nematode infections were collected during the study period. Results A total of 202 880 person-times were subjected to stool examinations in Zhenjiang City from 2006 to 2018, and 946 persons were detected with soil-transmitted nematode infections, with a mean prevalence of 0.47%. The overall prevalence of human soil-transmitted nematode infections appeared a decline tendency (P < 0.01), with a reduction from 1.10% in 2006 to 0.11% in 2018, and there was a region-specific overall prevalence (P < 0.01), with the highest prevalence seen in Runzhou District (1.65%) and the lowest detected in New District (0.04%). The mean prevalence of human Ascaris lumbricoides, hookworm and Trichuris trichiura infections was 0.33%, 0.09% and 0.04% in Zhenjiang City, respectively, and showed a decline tendency year by year from 2006 to 2018 (P < 0.01). Among all infections, mild infections were predominant. During the period from 2006 through 2018, a total of 45 427 children were examined in Zhenjiang City from 2006 to 2018, and 145 children were positive for E. vermicularis infections, with a mean prevalence of 0.32%. The prevalence of E. vermicularis infections decreased from 1.33% in 2006 to 0.22% in 2018 in children, and the overall prevalence appeared a decline tendency (P < 0.01). A single parasite infection was predominant in soil-transmitted nematode infections (97.46%), and mixed infection of two parasites were mainly detected before 2007. A total of 535 089 person-times received deworming in Zhenjiang City from 2006 to 2018, and the rate of adverse events was 0.002%. Conclusions Currently, the soil-transmitted nematode infection is at a low level in Zhenjiang City, and the overall prevalence of soil-transmitted nematode infections shows a decline tendency year by year. The surveillance and health education of soil-transmitted nematodiasis should be further intensified to consolidate the control achievements.
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Human genetic resources are valuable for life science research and pharmaceutical industry.Competitions for human genetic resources and the relevant techniques and industries have increasingly become intense among countries with the the implementation of precision medicine strategy and the maturity of gene editing technology.In the context of scientific progress and efficiency,the Organisation for Economic Co-operation and Development,the most important international economic organization,has proposed solutions to technological development and research paradigm changes from the perspective of national and global public interests.The United States,Japan,the United Kingdom,and some other developed countries have also released their policies and guidelines on the sharing of genetic resource data.In this article we analyzed these policies and guidelines,with an attempt to further improve the administration of human genetic resource sharing in China and promote the legal sharing and effective use of these resources.
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Humans , China , Guidelines as Topic , Human Genetics , Information DisseminationABSTRACT
Background@#On diagnosis of human immunodeficiency virus (HIV) infection, a person may have been infected already for many years. This study aimed to estimate the duration of HIV infection at the time of diagnosis.@*Methods@#Newly diagnosed HIV cases in Dehong, China, from 2008 to 2015 were studied. Duration of infection at the time of diagnosis was calculated using the first CD4 cell count result after diagnosis and a CD4 depletion model of disease progression. Multiple linear regression analysis was used to investigate the associated risk factors.@*Results@#A.@*@#total of 5867 new HIV cases were enrolled. Overall, mean duration of infection was 6.3 years (95% confidence interval [CI]: 6.2, 6.5). After adjusting for confounding, significantly shorter durations of infection were observed among participants who were female (beta: -0.37, 95% CI: -0.64, -0.09), Dai ethnicity (beta: -0.28, 95% CI: -0.57, 0.01), and infected through injecting drug use (beta: -1.82, 95% CI: -2.25, -1.39). Compared to the hospital setting, durations were shorter for those diagnosed in any other settings, and compared to 2008, durations were shorter for those diagnosed all years after 2010.@*Conclusion@#Although the reduction in duration of infection at the time of diagnosis observed in Dehong was significant, it may not have had a meaningful impact.
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Adult , Female , Humans , Male , Middle Aged , Young Adult , CD4 Lymphocyte Count , China , HIV Infections , Diagnosis , Risk Factors , Substance-Related Disorders , Time FactorsABSTRACT
To explore potent anticancer agent based on artemisinin scaffold, a series of 10--phenyl ethers derivatives containing dihydropyrazolyl or pyrazolyl moiety have been designed and synthesized. Their structures were determined by LC-MS and ¹H-NMR date. Inhibitory effects of the target compounds in human breast cancer MCF-7, MCF/Adr, MDA-MB-231 cells and prostate cell line PC-3 were determined by MTT assay. Those derivatives displayed good antiproliferative activity against the tested cancer cells. Particularly, target compounds exhibited significant cytotoxicity against drug-resistance cells MCF/Adr, which was worthy for further investigation.
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Objective To analyze the epidemic situation and epidemiological characteristics of malaria in Zhenjiang City from 2004 to 2017, so as to provide the evidence for formulating the reasonable strategies and measures for malaria prevention and control. Methods The epidemiological data of malaria in Zhenjiang City from 2004 to 2017 were collected and analyzed for the epidemic situation, epidemiological characteristics, species of Plasmodium, blood test status for febrile patients. Results A total of 182 malaria cases were reported in Zhenjiang City between 2004 and 2017, 123 cases (67.58%) were infected with P. vivax, and 55 cases (30.22%) were infected with P. falciparum. Among all the reported cases, 41 (22.53%) were local cases, 66 (36.26%) were imported cases from other provinces in China, 75 (41.21%) were imported cases from abroad. Most of the cases were male (133 cases), and 59.89% of them concentrated in the age groups of 20-50 years. These cases were mainly farmers (24.73%, 45/182) and workers (24.18%, 44/182), and most of the cases occurred from July to September. From 2004 to 2017, the blood tests were conducted for 258 879 febrile patients, with a positive detection rate of 0.007% in the local population and 0.724% in floating population, and there was a statistically significant difference between them (P<0.01). Conclusion Since 2011, no indigenous cases have been reported in Zhenjiang City, and therefore, the control of imported malaria from abroad has become the key of malaria prevention and control in this city.
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Objective To analyze the epidemic situation and epidemiological characteristics of malaria in Zhenjiang City from 2004 to 2017, so as to provide the evidence for formulating the reasonable strategies and measures for malaria prevention and control. Methods The epidemiological data of malaria in Zhenjiang City from 2004 to 2017 were collected and analyzed for the epidemic situation, epidemiological characteristics, species of Plasmodium, blood test status for febrile patients. Results A total of 182 malaria cases were reported in Zhenjiang City between 2004 and 2017, 123 cases (67.58%) were infected with P. vivax, and 55 cases (30.22%) were infected with P. falciparum. Among all the reported cases, 41 (22.53%) were local cases, 66 (36.26%) were imported cases from other provinces in China, 75 (41.21%) were imported cases from abroad. Most of the cases were male (133 cases), and 59.89% of them concentrated in the age groups of 20-50 years. These cases were mainly farmers (24.73%, 45/182) and workers (24.18%, 44/182), and most of the cases occurred from July to September. From 2004 to 2017, the blood tests were conducted for 258 879 febrile patients, with a positive detection rate of 0.007% in the local population and 0.724% in floating population, and there was a statistically significant difference between them (P<0.01). Conclusion Since 2011, no indigenous cases have been reported in Zhenjiang City, and therefore, the control of imported malaria from abroad has become the key of malaria prevention and control in this city.
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A retrospective surveillance study on enterovirus D68 was performed in Beijing, China, following the largest and most widespread EV-D68 infection, which occurred in the USA. From January 2011 to July 2015, EV-D68 was identified in 12 individuals with respiratory infections in Beijing, China. The phylogenetic relationships based on the genomic sequence alignment showed that there were two lineages circulating in Beijing from 2011 to 2015. Eight EV-D68 strains belonged to group 1 and four belonged to group 3. All EV-D68 strains from Beijing in 2014 were separately clustered into subgroup II of group 1. Based on these results, we concluded that the Beijing EV-D68 strains had little association with the EV-D68 strains circulating in the 2014 USA outbreak.
Subject(s)
Adolescent , Aged , Child , Child, Preschool , Female , Humans , Male , Beijing , Epidemiology , Enterovirus D, Human , Classification , Genetics , Enterovirus Infections , Epidemiology , Virology , Genome, Viral , Phylogeny , Retrospective StudiesABSTRACT
No direct comparison of tauroursodeoxycholic acid (TUDCA) and ursodeoxycholic acid (UDCA) has yet been carried out in the treatment of liver cirrhosis in China. We designed a double-blind randomized trial to evaluate the potential therapeutic efficacy of TUDCA in liver cirrhosis, using UDCA as parallel control. The enrolled 23 patients with liver cirrhosis were randomly divided into TUDCA group (n=12) and UDCA group (n=11), and given TUDCA and UDCA respectively at the daily dose of 750 mg, in a randomly assigned sequence for a 6-month period. Clinical, biochemical and histological features, and liver ultrasonographic findings were evaluated before and after the study. According to the inclusion criteria, 18 patients were included in the final analysis, including 9 cases in both two groups. Serum ALT, AST and ALP levels in TUDCA group and AST levels in UDCA group were significantly reduced as compared with baseline (P<0.05). Serum albumin levels were significantly increased in both TUDCA and UDCA groups (P<0.05). Serum markers for liver fibrosis were slightly decreased with the difference being not significant in either group. Only one patient in TUDCA group had significantly histological relief. Both treatments were well tolerated and no patient complained of side effects. It is suggested that TUDCA therapy is safe and appears to be more effective than UDCA in the treatment of liver cirrhosis, particularly in the improvement of the biochemical expression. However, both drugs exert no effect on the serum markers for liver fibrosis during 6-month treatment.
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No direct comparison of tauroursodeoxycholic acid (TUDCA) and ursodeoxycholic acid (UDCA) has yet been carried out in the treatment of liver cirrhosis in China. We designed a double-blind randomized trial to evaluate the potential therapeutic efficacy of TUDCA in liver cirrhosis, using UDCA as parallel control. The enrolled 23 patients with liver cirrhosis were randomly divided into TUDCA group (n=12) and UDCA group (n=11), and given TUDCA and UDCA respectively at the daily dose of 750 mg, in a randomly assigned sequence for a 6-month period. Clinical, biochemical and histological features, and liver ultrasonographic findings were evaluated before and after the study. According to the inclusion criteria, 18 patients were included in the final analysis, including 9 cases in both two groups. Serum ALT, AST and ALP levels in TUDCA group and AST levels in UDCA group were significantly reduced as compared with baseline (P<0.05). Serum albumin levels were significantly increased in both TUDCA and UDCA groups (P<0.05). Serum markers for liver fibrosis were slightly decreased with the difference being not significant in either group. Only one patient in TUDCA group had significantly histological relief. Both treatments were well tolerated and no patient complained of side effects. It is suggested that TUDCA therapy is safe and appears to be more effective than UDCA in the treatment of liver cirrhosis, particularly in the improvement of the biochemical expression. However, both drugs exert no effect on the serum markers for liver fibrosis during 6-month treatment.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Cholagogues and Choleretics , Therapeutic Uses , Double-Blind Method , Liver Cirrhosis , Diagnosis , Drug Therapy , Taurochenodeoxycholic Acid , Therapeutic Uses , Treatment Outcome , Ursodeoxycholic Acid , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To investigate the epidemiological characteristics of measles cases of new genotype D8 in Beijing from January to June, 2013.</p><p><b>METHODS</b>Epidemiological survey and descriptive analysis was conducted.</p><p><b>RESULTS</b>661 suspected measles were reported from January to June, 2013. 416 were confirmed measles cases by serology and etiology detection. 28 measles cases were caused by genotype D8 measles virus by genotype identification. There were 2 measles outbreak including 14 cases and 14 sporadic cases. The incidence peak was during April and May. 25 cases (89.3%, 25/28) occurred in downtown and suburban districts. 22 cases (78.5%, 22/28) were adults aged 15-39 years and 19 cases (67.9%, 19/28) were migrant population. 12 cases (85.7%, 12/14) in outbreak were migrant population working in clothing sales. There was epidemiological association between 2 outbreaks.</p><p><b>CONCLUSION</b>Measles cases of genotype D8 were found for the first time in Beijing. Genotype D8 virus mainly infected migrant adults and caused local outbreak and endemic.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , China , Epidemiology , Disease Outbreaks , Genotype , Incidence , Measles , Epidemiology , Virology , Measles virus , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To screen and identify the possible existence of natural antisense transcript (NAT) within the mouse neocortex.</p><p><b>METHODS</b>Sixty-three cerebral cortex layer-specific genes were screened by bioinformatics prediction in mice, among which 31 mice with potential NATs were screened. NAT was identified using reverse transcription polymerase chain reaction (RT-PCR) and then cloned in pGEM-T Vector System for sequencing.</p><p><b>RESULTS</b>Among 31 genes predicted using bioinformatics, 8 were proved to be NAT positive by RT-PCR.</p><p><b>CONCLUSIONS</b>NATs exist in the mouse neocortex tissue during the development of cerebral cortex. NATs may influence mouse cortical development by regulating the related coding genes.</p>
Subject(s)
Animals , Mice , Cell Line , Cerebral Cortex , Molecular Sequence Data , RNA, Antisense , Genetics , RNA, Messenger , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To assess the expression level of D-Tyr-tRNA(Tyr) deacylase (DTD) in SAMP8 mice and speculate the function of DTD in disorders associated with Alzheimer's disease (AD).</p><p><b>METHODS</b>Altogether 12 SAMP8 mice and 12 SAMR1 mice were used in this study. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to detect the mRNA and protein levels of DTD in the mice. Purified DTD protein was injected into lateral ventricle to investigate the function of DTD in SAMP mice. The behavior of the mice was tested by using a Step-through Test System.</p><p><b>RESULTS</b>Both mRNA and protein levels of DTD were found to be significantly lower in SAMP8 mice compared with those in SAMR1 mice (P<0.05). In vivo injection of DTD protein did not lead to an obvious change in behavior of SAM mice.</p><p><b>CONCLUSIONS</b>DTD might function in the process of AD-associated pathology and could possibly participate in physiology process in a long-term manner to orchestrate with other regulators in order to maintain the balance of organism.</p>
Subject(s)
Animals , Mice , Alzheimer Disease , Aminoacyltransferases , Metabolism , Base Sequence , DNA Primers , Disease Models, Animal , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To evaluate the clinical function and significance of establishing a regional active neonatal transport network (ANTN) in Beijing.</p><p><b>METHOD</b>The authors retrospectively studied intensive care and the role of ANTN system in management of critically ill neonates and compared the outcome of newborn infants transported to our NICU before and after we established standardized NICU and ANTN system (phase 1: July 2004 to June 2006 vs phase 2: July 2006 to May 2008).</p><p><b>RESULT</b>The number of neonatal transport significantly increased from 587 during phase 1 to 2797 during phase 2. Success rate of transport and the total cure rate in phase 2 were 97.85% and 91.99% respectively, which were significantly higher than those in phase 1 (94.36% and 88.69%, respectively, P < 0.01). The neonatal mortality significantly decreased in phase 2 compared with that in phase 1 (2.29% vs 4.31%, P < 0.01). The capacity of our NICU was enlarged following the development of ANTN. There are 200 beds for level 3 infants in phase 2, but there were only 20 beds in phase 1. Significantly less patients in the phase 2 had hypothermia, acidosis and the blood glucose instability than those in phase 1 (P < 0.01, 0.05, 0.01 and 0.05, respectively). The proportion of preterm infants transported to our NICU were higher in phase 2 compared with that in phase 1, especially infants whose gestational age was below 32 weeks. The proportions of asphyxia and respiratory distress syndrome were lower in phase 2 than that in phase 1, but the total cure rates of these two diseases had no significant changes between the two phases. The most important finding was that the improvement of outcome of premature infants and those with asphyxia and aspiration syndrome was noted following the development of ANTN.</p><p><b>CONCLUSION</b>Establishing regional ANTN for a tertiary hospital is very important to elevate the total level in management of critically ill newborn infants. It plays a very important role in reducing mortality and improving total outcomes of newborn infants. There are still some problems remained to solve after four years practice in order to optimize the ANTN to meet needs of the development of neonatology.</p>
Subject(s)
Humans , Infant, Newborn , Infant Mortality , Infant, Premature , Intensive Care Units, Neonatal , Reference Standards , Transportation of Patients , Reference StandardsABSTRACT
<p><b>OBJECTIVE</b>To investigate possible relationship between expression of surfactant protein B (SP-B) gene product and neonatal respiratory distress syndrome (NRDS) in Han ethnic group.</p><p><b>METHOD</b>Unrelated 20 cases with NRDS of Han ethnic group were selected as NRDS group while unrelated 20 diseases cases of Han ethnic group with diseases were selected as control group. The cases in the control group had congenital heart disease or bronchopulmonary dysplasia or persistent pulmonary hypertension. Blood sample was taken from every case. Lung tissues were taken from the patients who died half an hour after death in the two groups. Expression of SP-B in lung tissue was determined with immunohistochemical tecnique. Genetic deficiency variant of SP-B intron IV was screened with polymerase chain reaction (PCR).</p><p><b>RESULTS</b>Two cases at gestational age 26 weeks and one case at gestational age 34 weeks and two cases at gestational age 42 weeks of NRDS groups had lower level expression of SP-B in lung tissue than those at the same age of NRDS. Expression of SP-B in lung tissue of control group increased with gestational age, but no such phenomenon was found in NRDS group. Further, two cases at gestational age 42 weeks of NRDS group had genetic deficiency variant of SP-B intron IV with gene analysis of five cases who had lower expression of SP-B. Clinical data suggest that patients at 42 weeks of gestational age had severe illness.</p><p><b>CONCLUSIONS</b>Decrease of SP-B expression may participate in occurrence of NRDS, genetic deficiency variant of SP-B intron IV exists in the NRDS cases of Han ethnic group of China.</p>
Subject(s)
Humans , Infant, Newborn , Bronchopulmonary Dysplasia , Genetics , China , Ethnicity , Genetics , Genetic Predisposition to Disease , Genome-Wide Association Study , Gestational Age , Introns , Polymorphism, Genetic , Pulmonary Surfactant-Associated Protein B , Genetics , Pulmonary Surfactants , Therapeutic Uses , Respiratory Distress Syndrome, Newborn , Genetics , WillsABSTRACT
The aim of this study is to construct a lentiviral vector encoding human growth hormone, and to achieve the long, efficient and stable expression in murine skeletal myoblasts. Primary skeletal myoblasts were isolated from Sprague-Dawley rats and cultured by enzymatic digestion. We tested them by Desmin immunohistochemistry stains and found their viability was up to 94% by Trypan blue. Human growth hormone (hGH) cDNA was subcloned into expression vector pLenti6/V5-D-TOPO to construct recombinant pLenti6/V5-hGH. The pLenti6/V5-hGH and the contructed pLenti6/V5-EGFP were transfected into murine skeletal myoblasts by the Lipofectamin 2000. Through counting by the Confocal Laser Scanning Microscope, we identified the transfection efficency. We added the blasticidin to the 6-well plate with lids and obtained stable myoblasts expressing hGH. The concentration of human growth hormone (hGH) in cell culture medium was detected by Radioimmunoassay (RIA). Polymerase Chain Reaction (PCR) and DNA sequence showed hGH cDNA had been correctly inserted into pLenti6/V5-D-TOPO vector. Bright green fluorescence of the transfected cells could be observed under the Confocal Laser Scanning Microscope after 24 h transfection with pLenti6/V5-EGFP plasmids, and the transfection rate reached 40%. The difference was distinct (P < 0.01) between the pLenti6/V5- hGH groups and control groups in the secretive level of human growth hormone. After 8 weeks, the expression of human growth hormone was still stable. Then, we validated the biological characterization of the rhGH by the enzyme-link immunosorbent assay (ELISA) of the Insulin-like growth factor I (IGF-1). These results demonstrate we have successfully constructed the recombinant pLenti6/V5-hGH plasmids and accomplished rhGH long, efficient and stable expression ectopic in skeletal muscle myoblasts.
Subject(s)
Animals , Humans , Rats , Animals, Newborn , Cells, Cultured , Genetic Vectors , Human Growth Hormone , Genetics , Lentivirus , Genetics , Metabolism , Myoblasts, Skeletal , Metabolism , Plasmids , Genetics , Rats, Sprague-Dawley , Recombinant Proteins , Genetics , TransfectionABSTRACT
Objective To investigate the developmental characters of neural stem cells(NSCs) in occipital of cortex from human fetal brain at different age.Methods Ninety cases of embryoes at gestational age 16-32 weeks and by induction of labor with water bag were collected for determining distribution,shapes,growth modes and the number of NSCs in the occipital of cortex with immunohisto- chemical method under light microscope.Results It was noted that NSCs existed in the occipital of cortex from human fetal brain at different ages.NSCs mainly distributed in layers of cone cells and inner granule cells.NSCs existed in the occipital of cortex of different fetal age included middling round cells,NSCs had enations from 0 to 1.Nucli were larger than plasm.Each NSC had nucleoli from 2-4 and rarefaction chromatin.Most of NSCs distributed in three growth modes including crowd,cluster and clone,occasionally with a single growth mode among other nerve cells.There were no differences including distribution,shapes,growth modes and the number of NSCs in the occipital of cortex between groups,but,NSCs gradually decreased with increasing of age.Conclusion NSCs exists in the occipital of cortex from different gestational age,and the number of NSCs decreases with increasing of age.
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<p><b>BACKGROUND</b>The activation of extracellular signal-regulated kinase1/2 (ERK1/2) has been shown to be important signaling pathway in the ischemic preconditioning (IPC) response. Recently, some studies suggest a key role for the mitochondrial ATP-sensitive potassium channel (mKATP) as both a trigger and an end effector of acute and delayed protection of IPC. Hence, this study was undertaken to elucidate the relationship between mKATP and ERK1/2 in the delayed protection mechanism of anoxic preconditioning (APC).</p><p><b>METHODS</b>An APC model was established using cultured neonatal rat cardiomyocytes. Pharmacological agents [diazoxide, 5-hydroxydecanoate (5-HD), 2-mercaptopropionylglycine (MPG), and PD98059] were used to modulate mKATP and ERK1/2 activation. Cellular injury was evaluated by measuring cellular superoxide dismutase (SOD) activity, cell viability, and lactate dehydrogenase (LDH) release. The generation of cellular reactive oxygen species (ROS) and the activation of ERK1/2 were determined at different time points starting from the beginning of preconditioning with anoxia or diazoxide (an mKATP opener).</p><p><b>RESULTS</b>Cell viability and SOD activity in the APC [(81.9 +/- 11.4)%, (13.6 +/- 3.7) U/L] and diazoxide [(79.2 +/- 12.4)%, (16.5 +/- 4.6) U/L] groups were significantly higher than in the anoxia/reoxygenation (A/R) [(42.2 +/- 7.3)%, (8.8 +/- 2.8) U/L] group (all P < 0.01). LDH activity in the APC group [(101.9 +/- 18.9) U/L] and diazoxide group [(97.5 +/- 17.7) U/L] was significantly lower than in the A/R group [(250.5 +/- 43.6) U/L] (all P < 0.01). Both APC and diazoxide simultaneously facilitated intracellular ROS generation and rapid ERK1/2 activation. But the effects of APC and diazoxide were remarkedly attenuated by 5-HP (an mKATP blocker) and by MPG (a free radical scavenger). In addition, the ERK1/2 inhibitor PD98059 also abolished the cellular protective effects induced by diazoxide.</p><p><b>CONCLUSION</b>mKATP may mediate ERK1/2 activation during anoxia preconditioning by generating ROS, which then triggers the delayed protection of APC in rat cardiomyocytes.</p>