2.Construction of fusion expression vector pET22b-SUMO-FGFR4 and optimization of expression conditions in E.coli
Wei LIU ; Yang YAO ; Xiaoxiao MA ; Yuxuan DENG ; Di MEI ; Lei LIU ; Huiyan WANG
Journal of Jilin University(Medicine Edition) 2016;42(4):642-647
Objective:To design the small ubiquitin modification-fibroblast growth factor receptor 4 (SUMO-FGFR4) fusion gene and construct the expression vector pET22b-SUMO-FGFR4, to optimize the expression conditions. Methods:The SUMO-FGFR4 fusion gene was obtained by Overlap PCR and was connected to pET22b;the recombinant expression vector pET22b-SUMO-FGFR4 was obtained. The influence of lactose concentration, induction time,induction temperature,induction point and adding mode of lactose in the expression levels was observed,and the best induction condition was determined; then the solubility of recombinant protein was analyzed.Results:The SUMO-FGFR4 fusion protein was highly expressed,the molecular weight of the fusion protein was about 40 000 and it could bind with FGFR4 specific antibody.When the lactose concentration was 1.0 g·L-1 ,the induction time was 3 h,the induction temperature was 37℃,the value of A (600)was 0.8,the expression level was highest;but adding mode of lactose had no remarkable effect on the protein expression.The expression level of recombinant protein induced by lactose was higher than IPTG.SUMO-FGFR4 protein existed in a form of inclusion body.Conclusion:The SUMO-FGFR4 fusion protein is expressed successfully in this study while lactose is used as inducer and the best expression conditions are confirmed.
3.Construction of GJB2 mutations common in Chinese EGFP fusion protein vectors
Yanping ZHANG ; Yuanding ZHANG ; Lina LI ; Lei MA ; Yurui SUN ; Zonglin ZHANG ; Jinwei LIU ; Huiyan DENG ; Wei ZHU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;(16):724-727
Objective:To construct GJB2 gene mutaitons common in Chinese EGFP fusion protein vectors, and to search for better way to study the mechanism of deletion mutaitons in GJB2 gene. Method: Non-fusion protein vectors of 235delC, 299-300 del AT and 176 del 16 bp were first made by point mutaiton methods in vitro. Then expression part of the upper 3 mutations were amplified by PCR and the PCR products were cloned into TA cloning vector. After cutting by restriction enzymes EcoRI/BamHI, three deletion mutaions were inserted into pEG-FP-N1 vector. Sequencing was used to verify the validity of the fusion protein vectors. HEK293 cells were trans-fected with the recombinant DNA samples by the liposome complex method. Results The recombined plasmids were highly expressed in HEK293 cells. Green fluorescence singals were distributed uniformly in cytoplasm. Conclusion; GJB2 mutations common in Chinese EGFP fusion protein vectors were constructed successfully. It may provide a better way to explore the reasons of nonsyndromic hearing loss common in Chinese.
4.The short-term effect of temperature on non-accidental mortality in Guangzhou, Changsha and Kunming.
Huiyan XIE ; Wenjun MA ; Yonghui ZHANG ; Tao LIU ; Hualiang LIN ; Jianpeng XIAO ; Yuan LUO ; Yanjun XU ; Xiaojun XU
Chinese Journal of Preventive Medicine 2014;48(1):38-43
OBJECTIVETo explore the relationship between temperature and non-accidental mortality in Guangzhou, Changsha and Kunming;to evaluate the temperature-related risk of mortality; and thereby to provide scientific evidence for enacting the policy to tackle climate changes.
METHODDaily meteorology data and mortality data were collected in 2006-2009 in Guangzhou, Changsha and Kunming. Distributed lag non-linear model (DLNM) was established and applied in a case-crossover design, which controlled the secular trend of time, to estimate the specified effects of temperature on non-accidental mortality at conditions of lag 0-2, lag 0-18 and lag 0-27 days, respectively.
RESULTAn obvious seasonal periodicity was found in non-accidental mortality in Guangzhou, Changsha and Kunming during 2006-2009. The mortality number was comparatively high in the winters, and some high temperature days in summer; but was comparatively low in springs and autumn. An L-shaped relationship was found between temperature and mortality in Guangzhou and Kunming and a U-shaped relationship was found in Changsha. When daily mean temperature exceeded 28.2 °C, 24.5°C and 23.2°C, as average temperature increase 1°C, non-accidental mortality increased 4.56% (95%CI:2.74%-6.63%), 5.66% (95%CI:0.22%-12.65%) , -3.94% (95%CI:-32.77%-39.01%) , respectively; when daily mean temperature below 24.8°C, 20.0°C and 17.3°C, as average temperature decrease 1°C, the corresponding increase in non-accidental mortality were 3.28% (95%CI:2.41%-4.10%) (lag 0-18 days), 1.35% (95%CI:0.31%-1.77%) (lag 0-2 days) and 2.42% (95%CI:1.08%-3.27%) (lag 0-27 days) , respectively. The effects of hot weather were acute and short term; while the effects of cold weather had a several days delay, but a longer persistence.
CONCLUSIONSExtreme cold and hot temperature could increase the risk of non-accidental mortality in Guangzhou, Changsha and Kunming. The effects of cold weather had a several days delay, but a longer persistence.
China ; epidemiology ; Climate Change ; Cross-Over Studies ; Humans ; Mortality ; Seasons ; Temperature
5.Nonfatal child pedestrian injury in two urban cities of Guangdong Province, China: results from a cross-sectional survey.
WenJun MA ; ShaoPing NIE ; HaoFeng XU ; YanJun XU ; HuiYan XIE ; YuRun ZHANG
Biomedical and Environmental Sciences 2011;24(4):335-342
OBJECTIVETo describe the epidemiological characteristics of nonfatal child pedestrian injuries and provide information to help understand an important public-health problem.
METHODSThis was a school-based, cross-sectional questionnaire survey. The sample (42 750 children) was obtained from two urban cities of Guangdong Province, China, using multi-stage randomized sampling. Information was collected by the respondents self-reporting in the classroom.
RESULTSThe incidence rate of nonfatal child pedestrian injuries in the cities was 2.0%. Boys had a higher incidence rate (2.6%) than girls (1.4%). Compared to other children, those aged 10 years are at the highest risk. The primary places of occurrence were sidewalks, residential roads, and crosswalks. High-risk behavior of the children immediately prior to injury included mid-block crossings, playing on roads, and crossing on red lights. The major vehicles that caused pedestrian injuries were bicycles, car or vans, and motorcycles. Bruises, fractures, and injuries to the internal organs were the top three types of injuries. Almost 40% of victims were hospitalized, and nearly 30% of the victims suffered long-term disabilities.
CONCLUSIONThis study shows that nonfatal child pedestrian injuries are a very serious public-health problem in the urban cities of Guangdong. Based on the epidemiological characteristics, prevention strategies and further research should be carried out to reduce the occurrence of injuries.
Accident Prevention ; methods ; Accidents, Traffic ; prevention & control ; statistics & numerical data ; Animals ; Child ; China ; epidemiology ; Cross-Sectional Studies ; Data Collection ; Female ; Humans ; Incidence ; Male ; Motor Vehicles ; Surveys and Questionnaires ; Urban Population ; statistics & numerical data ; Wounds and Injuries ; epidemiology
6.Study on the Safety and Influencing Factors of Home Medication for Elderly Patients with Chronic Diseases in Urban Areas of Henan:A Cross-sectional Survey
Huiyan MA ; Zhigang ZHAO ; Chunfeng QIAO ; Shu GE ; Boya ZHOU ; Xixi LI ; Mingfen WU
Herald of Medicine 2024;43(12):1951-1957
Objective To evaluate the status of home medication safety among elderly patients with chronic diseases and analyze its influencing factors in urban and rural areas of Henan Province.Methods Convenient sampling method was adopted.Data was collected through a designed and optimized questionnaire.Pharmacists conducted in-home surveys.Excel and SPSS 26.0 software were used for data analysis.Results A total of 352 valid questionnaires were analyzed.Most respondents were aged 60~70 years,with a female proportion of 56.2%,and 52.0%of them had a junior high school education or lower.The top-ranked chronic diseases were hypertension(62.2%)and hyperlipidemia(33.2%),and 61.1%of patients suffered from multiple chronic diseases.Antipyretic and analgesic drugs were most common in home medicine cabinets(90.1%).The proportion of drugs being stored in accordance with the instructions(46.9%)was slightly low,and 93.8%of the elderly handled expired drugs improperly.Awareness of medication guidance services was low(34.9%).Logistic regression analysis showed that factors such as education level and monthly income had significant effects on the understanding of drug package insert,rational drug storage,proper disposal of expired drugs,and awareness of medication guidance services(P<0.05).Conclusion There are still some potential safety risks in the home medication use of elderly patients with chronic diseases in urban and rural areas of Henan Province.It is necessary to enhance education on drug storage and medication guidance,particularly for low-income and less-educated groups,to improve the safety of home medication use.
7. Association between both maternal pre-pregnancy body mass index/gestational weight gain and overweight/obesity children at preschool stage
Jingjing LIANG ; Yan HU ; Yanfei XING ; Ying MA ; Lin JIANG ; Huiyan LIU ; Fang HU ; Shaomin LU ; Suifang LIN
Chinese Journal of Epidemiology 2019;40(8):976-981
Objective:
To investigate the association of both maternal pre-pregnancy body mass index (BMI) and gestational weight gain (GWG) with childhood overweight and adiposity in preschool children.
Methods:
A total of 4 303 preschool children aged 3-5 years were enrolled in our study during June and November 2016 in Guangzhou. Children defined as overweight and obesity were according to the criteria of WHO while weight status during maternal pre-pregnancy was using the China Adult Reference. Gestational weight gain was defined according to the Institute of Medicine guidelines.
Results:
After adjusting the possible confounding factors, results from the logistic regression analysis showed that both maternal pre-pregnancy overweight and obesity would increase the risk for both childhood overweight and obesity (
8.Construction of GJB2 mutations common in Chinese EGFP fusion protein vectors.
Yanping ZHANG ; Yuanding ZHANG ; Lina LI ; Lei MA ; Yurui SUN ; Zonglin ZHANG ; Jinwei LIU ; Huiyan DENG ; Wei ZHU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;23(16):724-727
OBJECTIVE:
To construct GJB2 gene mutations common in Chinese EGFP fusion protein vectors, and to search for better way to study the mechanism of deletion mutations in GJB2 gene.
METHOD:
Non-fusion protein vectors of 235delC, 299-300 del AT and 176 del 16 bp were first made by point mutation methods in vitro. Then expression part of the upper 3 mutations were amplified by PCR and the PCR products were cloned into TA cloning vector. After cutting by restriction enzymes EcoRI/BamHI, three deletion mutations were inserted into pEGFP-N1 vector. Sequencing was used to verify the validity of the fusion protein vectors. HEK293 cells were transfected with the recombinant DNA samples by the liposome complex method.
RESULT:
The recombined plasmids were highly expressed in HEK293 cells. Green fluorescence signals were distributed uniformly in cytoplasm.
CONCLUSION
GJB2 mutations common in Chinese EGFP fusion protein vectors were constructed successfully. It may provide a better way to explore the reasons of nonsyndromic hearing loss common in Chinese.
Asian Continental Ancestry Group
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genetics
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Connexin 26
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Connexins
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genetics
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Genetic Vectors
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Green Fluorescent Proteins
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genetics
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Humans
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Sequence Deletion
9.Value of ultrasonic measurement of the ratio of optic nerve sheath diameter to eyeball transverse diameter in the diagnosis and prognosis of intracranial hypertension in patients with craniocerebral trauma
Kun ZHANG ; Fengjie MA ; Huiyan LI ; Yayun FANG ; Chao LONG ; Ran LIU ; Liping SONG
Chinese Journal of Postgraduates of Medicine 2024;47(2):134-138
Objective:To investigate the value of ultrasonic measurement of the ratio of optic nerve sheath diameter (ONSD) to eyeball transverse diameter(ETD) in the diagnosis and prognosis of intracranial hypertension in patients with craniocerebral trauma.Methods:A total of 120 patients with craniocerebral trauma treated in the Xingtai General Hospital of North China Medical and Health Group from December 2021 to January 2023 were perspectively selected, and they were divided into normal intracranial pressure group (73 cases) and intracranial hypertension group (47 cases) according to the results of intracranial pressure measurements, and the intracranial hypertension group was divided into good prognosis group (20 cases) and poor prognosis group (27 cases) according to the follow-up prognosis. The efficacy of ONSD, ETD and ONSD/ETD in intracranial hypertension diagnosis and prognosis assessment were analyzed by receiver operating characteristic (ROC) curve. Kaplan-Meier method was used to evaluate the 6-month risk of adverse prognosis of patients, and the comparison was made by Log-rank test.Results:The levels of intracranial pressure, ONSD, ONSD/ETD in the normal intracranial pressure group were lower than those in the intracranial hypertension group: (130.73 ± 23.63) mmH 2O (1 mmH 2O = 0.009 8 kPa) vs. (270.11 ± 35.78) mmH 2O, (5.47 ± 0.29) mm vs. (5.78 ± 0.44) mm, 0.246 ± 0.018 vs. 0.263 ± 0.018, there were statistical differences ( P<0.05). The scores of Glasgow Coma Scale (GCS), intracranial pressure, ONSD, ONSD/ETD in the good prognosis group were lower than those in the poor prognosis group: (5.50 ± 1.24) scores vs. (6.41 ± 1.34) scores, (256.15 ± 30.23) mmH 2O vs. (280.44 ± 36.56) mmH 2O, (5.62 ± 0.40) mm vs. (5.90 ± 0.44) mm, 0.254 ± 0.014 vs. 0.270 ± 0.017, there were statistical differences ( P<0.05). ROC curve analysis results showed that the area under the curve (AUC) of ONSD and ONSD/ETD for diagnosing intracranial hypertension in patients with craniocerebral trauma were 0.718 and 0.765, respectively, and the critical values were 5.87 mm and 0.263, respectively. The AUC of ONSD and ONSD/ETD predicting prognosis of intracranial hypertension patients was 0.677 and 0.763, respectively, and the critical values were 5.90 mm and 0.267, respectively. Grouped by the threshold of ONSD/ETD for the prognosis of intracranial hypertension (0.267), the incidence of adverse prognosis in ONSD/ETD > 0.267 group was higher than that in the ONSD/ETD≤0.267 group, there was statistical difference ( P<0.05). Conclusions:ONSD/ETD can be used as an index for diagnosis and prognosis of intracranial hypertension.
10.Analysis of copy number variation in AZF region of Y chromosome in patients with spermatogenic failure.
Hui GAO ; Lijuan WANG ; Yaqin SONG ; Di MA ; Rui NIE ; Yuhua HU ; Huiyan HE ; Ruanzhang ZHANG ; Shayan WANG ; Hui GUO
Chinese Journal of Medical Genetics 2023;40(9):1068-1074
OBJECTIVE:
To explore the characteristics of copy number variation (CNV) within the Y chromosome azoospermia factor (AZF) region in patients with spermatogenesis disorders in the Shenzhen area.
METHODS:
A total of 123 patients with spermatogenesis disorders who had visited Shenzhen People's Hospital from January 2016 to October 2022 (including 73 patients with azoospermia and 50 patients with oligozoospermia) and 100 normal semen males were selected as the study subjects. The AZF region was detected with multiplex ligation-dependent probe amplification (MLPA), and the correlation between the CNV in the AZF region and spermatogenesis disorders was analyzed using the chi-square test or Fisher's exact test.
RESULTS:
19 CNV were detected among 53 patients from the 223 samples, including 20 cases (27.40%, 20/73) from the azoospermia group, 19 cases (38%, 19/50) from the oligozoospermia group, and 14 cases (14%, 14/100) from the normal control group. In the azoospermia, oligozoospermia, and normal control groups, the detection rates for CNV related to the AZFa region (including AZFab and AZFabc) were 5.48% (4/73), 2.00% (1/50), and 0 (0/100), respectively. The detection rates for the AZFb region (including the AZFbc region) were 6.85% (5/73), 0 (0/50), and 0 (0/100), respectively. The detection rates for gr/gr deletions in the AZFc region were 2.74% (2/73), 6.00% (3/50), and 9.00% (9/100), respectively, and those for b2/b4 deletions in the AZFc region were 2.74% (2/73), 10.00% (5/50), and 0 (0/100), respectively. The detection rates for complex rearrangements in the AZFc region were 6.85% (5/73), 18.00% (9/50), and 3.00% (3/100), respectively. Statistical analysis showed no significant difference in the detection rate of gr/gr deletions between the three groups (Fisher's Exact Test value = 2.712, P = 0.249); the differences in the detection rate of b2/b4 deletions between the three groups were statistically significant (Fisher's Exact Test value = 9.489, P = 0.002); the differences in the detection rate of complex rearrangements in the AZFc region between the three groups were statistically significant (Fisher's Exact Test value = 9.493, P = 0.006). In this study, a rare AZFa region ARSLP1 gene deletion (involving SY86 deletion) was detected in a patient with oligozoospermia.
CONCLUSION
CNV in the AZFa and AZFb regions have a severe impact on spermatogenesis, but partial deletion in the AZFa region (ARSLP1 gene deletion) has a minor impact on spermatogenesis. The b2/b4 deletion and complex rearrangement in the AZFc region may be risk factors for male infertility. The gr/gr deletion may not serve as a risk factor for male infertility in the Shenzhen area.
Humans
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Male
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Azoospermia/genetics*
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DNA Copy Number Variations
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Oligospermia/genetics*
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Infertility, Male/genetics*
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Y Chromosome