1.Expression and localization of transmembrane protein CMTM2 in human testis and sperm
Xiaowei ZHANG ; Ke LAN ; Wenbo YANG ; Qing LI ; Yongping ZHAO ; Huaqi YIN ; Brandes KITE ; Wenjun BAI ; Tao XU
Journal of Peking University(Health Sciences) 2017;49(4):575-579
Objective: To study the expression of transmembrane protein CMTM2 in the testis and sperm of adult males and to approach the potential function of the protein in the male reproductive system.Methods: The expression of CMTM2 in human testis and sperm was confirmed by Western blot.Immunohistochemical staining was used for detecting CMTM2 localization in the testis tissue, TRITC-CMTM2 and FITC-Hoechst double immunofluorescence staining was performed to examine the subcellular localization of CMTM2 in the human sperm before and after acrosome reaction, that is, immunofluorescent staining was used for detecting CMTM2 localization in both the testis and sperm before and after the acrosome reaction.Results: CMTM2 was presented in both human testis and sperm.In the testis, CMTM2 immunoreactive particles were observed mainly in the membrane of the different stages of spermatogenic cells.In the human sperm, its immunoreactivity was restrictively localized to the posterior head where sperm-egg fusion occurred, and the CMTM2 localization was not affected by sperm acrosome reaction.CMTM2 was widely expressed in seminiferous tubules of the human testis, mainly in the cell membranes of spermatogenic cells, which was consistent with the previous reports.The immunofluorescence performed on frozen human testis slides showed similar findings with immunohistochemistry, which gave weight to the localization of CMTM2 in the cell membranes of spermatogenic cells at different stages.TRITC-CMTM2 and FITC-Hoechst double immunofluorescence staining was performed to examine the subcellular localization of CMTM2 in the human sperm before and after acrosome reaction.CMTM2 was localized at the posterior head of sperm before and after acrosome reaction.The localization and expression of CMTM2 were not affected by sperm acrosome reaction.Conclusion: Expression of CMTM2 in the male reproductive system of the adult human exhibits cell-and region-specific patterns, which suggests that they may play an important role in spermatogenesis and sperm-egg fusion.The expression of CMTM2 in the male reproductive system of the adult human exhibits cell-and region-specific patterns, which suggests that they may play an important role in spermatogenesis and sperm-egg fusion.However, it still remains to be further elucidated about the definite role of CMTM2 in male reproductive system and the process of spermatogenesis.And in vitro fertilization experiments are needed to confirm the role of CMTM2 in fertilization in future.
2.Effect of occupational stress on cardiovascular function of different vocational population.
San-qiao YAO ; Xue-yun FAN ; Yu-lan JIN ; Yu-ping BAI ; Yin-e QU ; Yuan ZHOU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(1):20-22
OBJECTIVETo study the effect of occupational stress on cardiovascular function of different vocational population.
METHODSThe occupational stressors, risk factors of cardiovascular diseases were investigated by questionnaire in 839 people with 4 kinds of jobs. Blood pressure, sugar, and lipid were detected at the same time.
RESULTSBlood pressure were higher in the groups of old age, long standing and teachers, and the abnormal rate of blood pressure was 21.69%. There was no difference in abnormal ECG among ages, standing and occupation, and the abnormal rate of ECG was 19.07%. Job control, job demands, job responsibility, role in a job and shift work were the main stress factors affecting systolic and diastolic blood pressure. More conflict in job, less chance of participation, severe job loads were the risk factors of primary hypertension. Accident due to job responsibility, job responsibility, role in a job were the main risk factors of abnormal electrocardiograph. Self-respect and activity beyond work were the good modifiers of heart function.
CONCLUSIONOccupational stress has certain effect on cardiovascular function.
Adult ; Blood Pressure ; Electrocardiography ; Female ; Humans ; Logistic Models ; Male ; Occupational Diseases ; physiopathology ; Stress, Psychological ; physiopathology
3.Study on the change of urinary iodine of different salt concentrations among the target population in an intervention trial
Yin-Yin SONG ; Li-Hong MU ; Ge LI ; Gui-Wang DOU ; Xin-Shu LI ; Xin LIU ; Bai-Sheng QIAN ; Jian-Lan CHEN ; Mei LUO
Chinese Journal of Epidemiology 2011;32(6):576-578
Objective To analyze the change of urinary iodine in a cohort of intervention trial and to observe the role of different doses on salt iodization and related impact factors on nutritional condition of iodine. Methods Multistage cluster sampling was used to sample three townships in two counties for community intervention with different doses (15 ± 5, 25 ± 5, 35 ± 5)mg/kg. Results Compared to the (35 ± 5)mg/kg group, the urine iodine levels of three experimental townships were gradually declining in county B when time went on, and the (15 ± 5) mg/kg group showed anobvious results, at 6,12,18 and 24 months, with the urine iodine level as 180.00,186.10,150.04,191.28 μg/L respectively, which were in accordance with the WHO standard and reached to appropriate range (187.96μg/L) at the 18 month. The townships at county Y under intervention had declined slightly, but the urine iodine levels did not reach the WHO standard. The thyroid volume declined from 3.65 ml to 3.40 ml in two counties and the difference between them was statistically significant. Conclusion To some extent, reducing the iodine concentration in salt, had a role of lowering the urine iodine level and reducing the strumous rate.
4.Protective effect of medicated serum prepared with taohong siwu tang on hydrogen peroxide-induced human umbilical vein endothelial cells.
Zhu-Qing LIU ; Deng-Ke YIN ; Lan HAN ; Bai-Kun LI ; Dai-Yin PENG
China Journal of Chinese Materia Medica 2013;38(3):402-406
OBJECTIVETo study the protective effect of medicated serum prepared with Taohong Siwu Tang on hydrogen peroxide-injured human umbilical vein endothelial cells (HUVECs).
METHODSprague Dawley rats were orally administered with 10 mL x kg(-1) extracts from Taohong Siwu Tang (1.75 g crude drug), twice a day for three days, in order to prepare medicated serum of Taohong Siwu Tang. The effect of medicated serum pre-treated with Taohong Siwu Tang (with concentrations of 5%, 10%, 15%) on reduction of H2O2-induced cell activity was detected MTT. Cell morphological changes were observed under microscope. The effect of medicated serum prepared with Taohong Siwu Tang on the apoptosis and antioxidant capacity of HUVECs was detected with the content of malondialdehyde (MDA) and dehydrogenase (LDH), the activity of superoxide dismutase (SOD) and AO/EB staining. The expression of Caspase-3 was determined by western blot.
RESULTCompared with the blank serum control group, cell were significantly less active after being damaged by H2O2 (400 micromol x L(-1)). Medicated serum could significantly improve the SOD activity, reduce the levels of LDH and MDA, and inhibite the expression of Caspase-3. AO/EB staining and inverted microscope also showed that medicated serum prepared with Taohong Siwu Tang could reduce cell apoptosis induced by H2O2.
CONCLUSIONMedicated serum prepared with Taohong Siwu Tang has significant protective effect on HUVECs injured by H2O2.
Animals ; Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Survival ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Hydrogen Peroxide ; pharmacology ; L-Lactate Dehydrogenase ; metabolism ; Malondialdehyde ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serum ; chemistry ; Superoxide Dismutase
5.Value of serum gamma-glutamyl transpeptidase combined with direct bilirubin in the diagnosis of biliary atresia in infants.
Hai-Yan FU ; Rui-Qin ZHAO ; Ge-Lan BAI ; Chun-Lan YIN ; Run-Kai YIN ; Hai-Hua LI ; Wei-Na SHI ; Ya-Li LIU ; Li-Juan CHENG ; Xiao-Yun JIA ; Gui-Gui LI ; Shi-Guang ZHAO
Chinese Journal of Contemporary Pediatrics 2019;21(12):1198-1202
OBJECTIVE:
To study the value of serum gamma-glutamyl transpeptidase (GGT) combined with direct bilirubin (DB) in the diagnosis of biliary atresia.
METHODS:
A total of 667 infants with cholestasis who were hospitalized and treated from July 2010 to December 2018 were enrolled as subjects. According to the results of intraoperative cholangiography and follow-up, they were divided into biliary atresia group with 234 infants and cholestasis group with 433 infants. The two groups were compared in terms of age of onset, sex, and serum levels of total bilirubin (TB), DB, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acid (TBA), and GGT. A receiver operating characteristic (ROC) curve analysis was performed for indices with statistical significance, and the area under the ROC curve (AUC) and the optimal cut-off value for diagnosis were calculated.
RESULTS:
The biliary atresia group had a significantly younger age of onset than the cholestasis group (P<0.001). There were no significant differences in sex, ALT, and AST between the two groups (P>0.05), while the biliary atresia group had significantly higher serum levels of TB, DB, TBA, and GGT than the cholestasis group (P<0.05). GGT combined with DB had the highest AUC of 0.892 (95% confidence interval: 0.868-0.916) in the diagnosis of biliary atresia. At the optimal cut-off values of 324.0 U/L for GGT and 115.1 μmmol/L for DB, GGT combined with DB had a sensitivity of 79.8% and a specificity of 83.2% in the diagnosis of biliary atresia.
CONCLUSIONS
GGT combined with DB has high sensitivity and specificity in the diagnosis of biliary atresia and can be used as an effective indicator for diagnosis of biliary atresia in infants.
Alanine Transaminase
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Aspartate Aminotransferases
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Biliary Atresia
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diagnosis
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Bilirubin
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Humans
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Infant
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gamma-Glutamyltransferase
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blood
6.Comparison between genotyping and serological phenotyping in RhCE blood group.
Hua-you ZHOU ; Yin-ze ZHANG ; Qing-bao MENG ; Xu-hua BAI ; Cong-rong WANG ; Qiong CAO ; Jiong-cai LAN
Chinese Journal of Medical Genetics 2008;25(1):66-69
OBJECTIVETo genotype the RHCE gene of Hans, Xinjiang's Uigurs and Kazakstans in China, and to compare the results of RHCE genotyping with that of RhCc/Ee phenotyping.
METHODSRHCE genes of 98 Hans with RhD positive and 230 Hans, 72 Uigurs and 18 Kazakstans with RhD/RHD negative were genotyped with PCR-sequence specific primer (SSP) technique.
RESULTSThe results of RHE/RHe genotyping from samples with RhD positive and negative were in accord with that of phenotyping. It would result in 4.44% error using C-->G polymorphism at nt48 of RHCE gene to genotype RHCE, and 4.05% failure of detection using the 109 bp insertion to detectRHCE gene in Chinese Hans. The results of RHE/RHe genotyping in unrelated 72 Uigurs and 18 Kazakstans with RhD phenotype were consistent with that of phenotyping, and false positive and false negative were not found in genotyping in Uigurs and Kazakstans tested.
CONCLUSIONThe results of RHE/RHe and RHc genotyping were correct with PCR-SSP and accordant with that of phenotyping. Using the C48G polymorphism in exon 1 of RHCE to genotype RHC gene would result in false positive resulting from RHc mutation at this locus, and using the 109 bp insertion to genotype RHC gene would result in false negative because of the absence of the 109 bp. Therefore it is necessary to genotype RHC gene using more than two polymorphic loci.
Ethnic Groups ; genetics ; Genotype ; Humans ; Phenotype ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; blood ; genetics ; Serologic Tests ; methods
7.A comparative research on RHD gene structures of Chinese Han and Uigur population.
Hua-you ZHOU ; Xu-hua BAI ; Yin-ze ZHANG ; Cong-rong WANG ; Qiong CAO ; Jiong-cai LAN
Chinese Journal of Medical Genetics 2006;23(2):151-155
OBJECTIVETo research comparatively on the RHD gene structures in unrelated RhD negative individuals of Chinese Uigur and Han population.
METHODSThe upstream, downstream, hybrid box and 10 exons of RHD gene were detected with sequence specific primer-PCR technique.
RESULTSThe results showed the genotypes of RhD negative individuals to have the significant difference between Chinese Uigur and Han population, that 94.44% Uigur individuals were with RHD(-)/RHD(-) genotype but just 61.40% Han population were with this genotype(94.44% versus 61.40%, P<0.01); 2.78% Uigur individuals were with RHD(+)/RHD(-) genotype but 34.21% Han population were with this genotype(2.78% versus 34.21%, P<0.01). However, there was significantly no RHD(+)/RHD(+) genotype difference between Chinese Uigur and Han population(2.78% versus 4.39%, P>0.05). In 78 cases of RhD negative Chinese Hans with single RHD gene, of which the RHD gene structure showed that 53(67.95%) cases were RHD(1-10) allele(of 53 RHD(1-10) alleles, 14 alleles were unexpressed); 15(19.23%) were RHD-CE(2-9)-D(2) allele; 5(6.41%) cases were RHD-CE(2-7)-D(2) allele; 2(2.56%) were similar to RHD-CE(3-6)D allele; 1(1.28%) case was RHD-CE(5-6)-D allele; and 2(2.56%) were RHD-CE(6)-D or point mutation respectively. Of 2 RhD negative Chinese Uigurs with RhD(-)/RHD(+) genotype, one carried RHD(1-10) allele, another carried RHD-CE(2-9)-D(2) allele.
CONCLUSIONThe most frequently unexpressed RHD alleles were RHD-CE(2-9)-D(2), RHD(1-10) and RHD-CE(2-7)-D(2) respectively in Chinese Han population who carried single RHD allele with RHD(-) phenotype and RHD(+) genotype. It showed the confluent character of RH gene in Chinese Han and Uigur population that there existed unexpressed RHD-CE(2-9)-D(2) allele in Chinese Uigur nationality, which was infrequent in Chinese Uigur population but frequent in Chinese Han population.
Alleles ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Ethnic Groups ; ethnology ; genetics ; Exons ; genetics ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Population Groups ; Rh-Hr Blood-Group System ; genetics
8.The effects of three kinds of biochemical indices on evaluation of occupational stress.
San-qiao YAO ; Xue-sheng WANG ; Yu-ping BAI ; Xue-yun FAN ; Shu-jie ZHANG ; Yu-lan JIN ; Yin-hui PEI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(1):23-26
OBJECTIVETo study the feasibility of monoamine neurotransmitters, serum glucose, serum glycerinate and cholesterol as objective indices for evaluating occupational stress.
METHODSJob stressors, modifiers, job strains in 844 people with four kinds of occupation were investigated, and the concentration of monoamine neurotransmitters, glucose, glycerinate and cholesterol in blood were detected at the same time. The methods of multiple stepwise regression and covariance analysis were used to analyze the data.
RESULTSThere was close relationship between monoamine neurotransmitters and job stressors, the forecast of the equation of 5-hydroxytryptamine (5-HT), dopamine (DA) and norepinephrine (NE) was 0.7238, 0.5703, 0.4438 respectively, the critical values of them were 804.00, 226.00 and 275.00 ng/ml respectively. There was a little contribution of job stressors to the equation of glucose, glycerinate and cholesterol, the critical values were 6.40, 2.51 and 5.92 mmol/L respectively.
CONCLUSIONMonoamine neurotransmitters may be a direct objective evaluating indices. Sugar, glycerinate and cholesterol may be an indirect objective indices.
Adult ; Blood Glucose ; analysis ; Cholesterol ; blood ; Female ; Humans ; Male ; Middle Aged ; Neurotransmitter Agents ; blood ; Occupational Diseases ; metabolism ; Stress, Psychological ; metabolism
9.Study of aniline's penetration capability through rats' complete skin in vitro and normal lab gloves.
Feng FENG ; Bai-yin HE ; Xue-lan YE ; Chang-qing LIU ; You-liang XIE ; Xiao-ping LAI ; Ying-jun ZHOU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(9):683-685
OBJECTIVETo study the permeability of intact mouse abdominal skin to aniline and the protective capability of two typical lab gloves against aniline.
METHODSA Franz diffusion cell was used to perform in vitro transdermal absorption test and glove permeation test for aniline (0.102 mg/ml and 0.010 mg/ml). The permeabilities of intact mouse abdominal skin and gloves to aniline were measured by high performance liquid chromatography-diode array detection.
RESULTSThe transdermal penetration of the two concentrations of aniline followed zero order kinetics within 12 h, exhibiting total aniline permeabilities within 24 h of 51.71% and 48.31%, respectively. The absorption liquid had an aniline concentration of at least 18 µg/L. The medical disposable latex glove could not stop the penetration of 0.010 mg/ml aniline, but the industrial natural latex glove could.
CONCLUSIONThe penetration of 0.102 mg/ml and 0.010 mg/ml aniline through the mouse abdominal skin follows zero order kinetics within 12 h. The medical disposable latex glove cannot stop the penetration of 0.010 mg/ml aniline, but the industrial natural latex glove can.
Aniline Compounds ; pharmacokinetics ; toxicity ; Animals ; Gloves, Protective ; Mice ; Skin Absorption ; drug effects
10.Prokaryotic expression and identification of HPV16 E7 protein.
Yan-Yan JIA ; Yue-Lan YIN ; Chun-Guang BAI ; Hong FU ; Yun-Fei GAO ; Zhi-Ming PAN ; Xin-An JIAO
Chinese Journal of Virology 2012;28(1):51-56
HPV16 E7 fusion protein was expressed in E. coli BL21, and its applied value for HPV was evaluated. HPV16 E7 gene was amplified by PCR, and cloned into prokaryotic expression vector pGEX6p-1. The recombinant plasmid was transformed into E. coli BL21, and HPV16 E7 fusion was expressed through IPTG induction. The expressed product was analyzed by SDS-PAGE and Western blot, subsequently purified according to Glutathione Sepharose 4B purification procedure. An indirect ELISA with the purified fusion protein as the coating antigen was then established to detect E7 serum antibodies from mice immunized with recombinant Listeria monocytogenes delivering HPV16 E7. The results demonstrated that the soluble fusion protein was highly expressed at 25 degrees C after induction with 0.5 mM IPTG. Furthermore, the result of Western blot analysis showed that the fusion protein had good specific reaction with an anti-E7 monoclonal antibody. Indirect ELISA result confirmed that the fusion protein could detect the serum antibodies against E7 with a titer of 1:200. The expressed GST-E7 fusion protein was immunocompetent, which was useful in the research of E7 biological function and therapeutic vaccine.
Animals
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli
;
genetics
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Female
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Mice
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Mice, Inbred C57BL
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Papillomavirus E7 Proteins
;
biosynthesis
;
genetics
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isolation & purification
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Polymerase Chain Reaction
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Recombinant Proteins
;
biosynthesis
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isolation & purification