1.Diagnosis and surgical treatment of bile duct stone of caudate lobe
Xizeng HUI ; Haiyan ZHENG ; Zhaoyang QIN ; Provincial SHANDONG
Chinese Journal of Current Advances in General Surgery 2009;0(08):-
Objective: To explore the diagnosis and surgical treatment for cholelithiasis of cau- dal lobe of liver. Methods: The data of 98 patients with cholelithiasis of caudal lobe were reviewed. Results: 42 cases (43% ) got preoperative diagnosis. No cases died during operation.7cases had residual stones,the rate of which was 7.1% . The stones recurred in 10 cases and the rate was 12.3% . Conclusion: Understanding the disease, the preoperative image examination combining with introperative exploration are important to decrease the misdiagnosis and increase the rate of diagnosis. The surgical treatment should follow the principle of individualized therapy.
2.Modified baculovirus system for high expression of Bombyx mori bidensovirus NS1 in silkworm.
Guohui LI ; Mangmang LI ; Qian ZHOU ; Zhaoyang HU ; Qi TANG ; Qin YAO
Chinese Journal of Biotechnology 2015;31(4):591-602
To improve the expression of heterologous genes using baculovirus expression system, we constructed a novel shuttle vector based on the Bm-Bacmid. In the Bm-Bacmid, partial sequences of Chitinase and Cystein Protease were replaced with a tandem cassette of Cm and egfp through homologous recombination. Bombyx mori bidensovirus (BmBDV) ns1 under the control of polyhedrin promoter was inserted into the modified Bm-bacmid by transposition. For comparison, BmBDV ns1 under the control of polyhedrin promoter was also cloned in the wild type Bm-bacmid. The resulting Bm-bacmids were transfected into the cultured BmN cells to prepare recombinant virus to infect silkworms for expression of BmBDV ns1. Total proteins of hemocyte from infected silkworms were subjected to Western blotting and ELISA analysis. The yield of BmBDV NS1 1 with the modified vector was three times as much as that with the unmodified vector. The method to improve the yield of BmBDV NS1 in silkworms will facilitate the function and three-dimensional structure study of BmBDV NS1.
Animals
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Baculoviridae
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Bombyx
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virology
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Cell Line
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Chitinases
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Cysteine Proteases
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Genetic Vectors
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Insect Viruses
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Promoter Regions, Genetic
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Transfection
3.Expression of ns1 gene from Bombyx mori bidensovirus by a novel baculovirus expression system.
Guohui LI ; Peng WANG ; Mangmang LI ; Wu XU ; Zhaoyang HU ; Qin YAO
Chinese Journal of Biotechnology 2014;30(4):625-635
Baculovirus gene expression is the most popular method to make target protein in cultured insect cells. To fast determine the generation of recombinant virus in cultured cells, donor plasmid of pFastBacI was modified by introducing egfp cassette. In the modified vector, egfp cassette was under the control of ie1 promoter, and target gene cassette was under the control of polyhedron promoter. To evaluate the convenience of the genetically modified donor plasmid used in eukaryotic expression, ns1 gene from Bombyx mori bidensovirus was ligated into the donor plasmid to generate recombinant plasmid pFastBacI-[P(ie1)-egfp-sv40]-[P(polh)-ns1-sv40]. Then the plasmid was transformed into DH10B competent cells containing Bm-Bacmid vector to produce the final recombinant Bm-Bacmid with the help of transposase. The resulting recombinant Bm-Bacmid was transfected into BmN cells to generate recombinant virus, which was easily and rapidly judged by green fluorescent signal observed in BmN cells. After infection for 96 h, the BmN cells were harvested and the total protein extracted from the infected BmN cells was subjected to Western blotting analysis. The result showed that a specific protein band about 36 kDa was detected, indicating that NS1 protein was successfully expressed in the BmN cells. In conclusion, the expression of NS1 protein with the modified expression system is useful for further research on the function of NS1 protein.
Animals
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Baculoviridae
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genetics
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Bombyx
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virology
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Cell Line
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Cells, Cultured
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Gene Expression
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Genetic Vectors
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Plasmids
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Promoter Regions, Genetic
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Transfection
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Viral Nonstructural Proteins
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biosynthesis
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genetics
4.Impact of pelvic autonomic nerve preservation on urination and potency in total mesorectal excision
Xizeng HUI ; Shide SONG ; Haiyan ZHENG ; Zhaoyang QIN ; Long LI ; Zhiwei ZHENG ; Tao XIE ; Xianbin LIU
Chinese Journal of Urology 2010;31(7):489-492
Objective To study the effect of pelvic autonomic nerve preservation(PANP)on urination and sexual function in total mesorectal excision(TME). Methods Two hundred and forty cases of male rectal cancer patients,divided into the PANP who accept the pelvic autonomic nerve preservation in TME,and the control group of 120 patients who do not.The urination and sexual function were observed and compared.3-year-survival rate,local recurrence rates of the two groups were recorded. Results The urinary disorder rates,erective disorder rates and ejaculation disorder rates of PANP group were 30.8%,28.3%and 34.2%,while values of control group were 55.0%、60.0%and 62.5%.The difference between them had statistical significance(P<0.05).The 3-year-survival rate and local recurrence rate of PANP group were 9.4%and 75.0%.The 3-year-survival rate and local recurrence rate of control group were 9.0%and 65.0%.There was no significant difference between them(P>0.05). Conclusion The PANP technique in TME could improve the urinary and sexual function of male patients without affect the prognosis.
5.Rescuing Bombyx mori bidensovirus in BmN cells in vitro.
Miaomiao ZHANG ; Ying MA ; Xiaoli PAN ; Zhaoyang HU ; Guohui LI ; Yayun SI ; Yali XING ; Keping CHEN ; Qin YAO
Chinese Journal of Biotechnology 2015;31(1):86-95
Bombyx mori bidensovirus (BmBDV) has been identified as causing chronic densonucleosis in Bombyx mori specifically. The replication mechanism of BmBDV remains unknown. Its genome comprises two single stands DNA (VD1 and VD2). In order to rescue infectious virions in vitro, we obtained the total viral DNA extracted from the BmBDV-infected larvae midguts, subsequently cloned the full-length sequence of BmBDV genome fragments by PCR and constructed recombinant plasmids pMD18T-VD1 and pUC-VD2. The linear genome fragments were obtained by digesting recombinant plasmids with corresponding restriction enzymes, and then collectively transfected BmN cells by the method of liposome-embedding. We determined the replication of the virus gene by PCR with the template of demethylated total DNA extracted from the post-transfect BmN cells. Meanwhile, we collected the total proteins from the post-transfect BmN cells and the larvae midgut of feeding the post-transfect BmN cells to perform Western blotting analysis, and detected the expression of viral genes. Here we firstly confirm that infectious virions can be rescued in BmN cells by linear co-transfect method.
Animals
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Bombyx
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DNA, Viral
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Densovirus
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growth & development
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Larva
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Transfection
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Virion
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Virus Cultivation
6.The study on the relationship between hippocampus neuronal apoptosis and hippocampus synaptic plasticity in rats exposed to aluminum.
Xiaohan NIE ; Xiujun QIN ; Huifang ZHANG ; Pan KANG ; Zhaoyang LI ; Qiao NIU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2015;33(7):535-538
OBJECTIVETo investigate the effect of aluminum exposure on neuronal apoptosis of rats hippocampus and the correlation of and synaptic plasticity.
METHODSThere were 40 SPF grade SD rats which were randomly divided into four groups: the control group, the low dose group, the medium dose group and the high dose group, 10 rats in each group. The rats were daily gavaged with aluminum lactate for 30 days. The hippocampal fEPSPs in rat was measured by electrophysiological grapher and the neuronal apoptosis in hippocampus was detected by Flow cytometer. In addition, the relative expression of gene which includes caspase-3, 8, 9 was measured by Real-time PCR.
RESULTSCompared to the control group, the average of fEPSPs which after HFS 10, 20, 30, 40, 50, 60 min was decreased at different time point in the low dose group, the medium dose group and the high dose group (P < 0.05). Compared with the control group, the rate of apoptosis was significantly increased in the medium dose group and the high dose group (P < 0.05). Compared to the control group, the relative expression of caspase-3 in the medium dose group and the high dose group was significantly increased in Real-time PCR (P < 0.05), and the relative expression of caspase-8 in the high dose group was significantly increased (P < 0.05).
CONCLUSIONAluminum exposure may induced neuronal apoptosis in rats, and then affect hippocampal synaptic plasticity.
Aluminum ; toxicity ; Aluminum Compounds ; toxicity ; Animals ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Hippocampus ; cytology ; drug effects ; Lactates ; toxicity ; Neuronal Plasticity ; drug effects ; Neurons ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley
7.Construction of a 5-Lipoxygenase Transgenic Mice
Meiying ZHANG ; Honglian WU ; Wei YANG ; Zhaoyang LI ; Wanwei DONG ; Shenglai ZHOU ; Yang YU ; Wei WANG ; Xiangchuan Lü ; Ying QIN ; Zhihong ZHENG ; Luzeng WANG
Acta Laboratorium Animalis Scientia Sinica 2010;18(1):60-64
Objective To construct a 5-lipoxygenase (5-LO) transgenic mouse model of atherosclerosis.Methods Purified 5-LO fragment was injected into male pronucli and the firtilized eggs were transplanted into pseudopregnant mice.PCR and Southern blot were used to detect the genotype of DNA separated from the newborn mouse tail tissues.RT-PCR and Western blot analysis were used to detect the gene transcription and expression.Results PCR and Southern blot results showed that 7 of 25 mice were transgenic mice.Expression of 5-LO and FLAP was found in the bone marrow,spleen,kidney,and peritoneal cells.Results of RT-PCR and Western blot showed that No.9,20,24transgenic mice expressed a higher level of 5-LO and FLAP than those in the wild type C57BL/6 mice.The expression levels in bone marrow and peritoneal cells were significantly different(P<0.05).Conclusion A 5-LO transgenie mouse line has been established in this study and may be used for future study on the function of 5-LO gene.
8.Effect of occupational factors on pre-diabetes mellitus among iron and steel workers
Yajing LIAO ; Chuxuan XU ; Chongqi MA ; Zhenwei QIN ; Yajiao SU ; Hongru ZHU ; Xiaotong ZHANG ; Chan LI ; Xiaoming LI ; Zhaoyang WANG ; Juxiang YUAN ; Hongmin FAN
Chinese Journal of Epidemiology 2020;41(6):929-933
Objective:To investigate the prevalence of pre-diabetes mellitus (PDM) and the impact of occupation-related factors on PDM, among workers from a steel company in Tangshan city, Hebei province.Methods:Clustering sampling method was used to select a steel company and to carry out occupational health-related physical checkup programs for eligible workers who had working in this company for longer than one year. The study began in February and ended up in June, 2017. Workers who were with FPG level as ≤6.9 mmol/L, and free from diabetes, were selected as the subjects for this study. Questionnaires were used and physical examinations and FPG testing conducted.Results:The total number of subjects in this study was 4 173, of which 2 648 appeared as pre-diabetic, with the prevalence rate as 63.4 %. Increase of the PDM prevalence was in parallel with the length of service, among the workers. The risk for the pre-diabetes in those who worked more than 8 hours per day was 1.696 times higher than those who worked less than or equal to 8 h/d (95 %CI:1.517-1.937). Compared with those workers without exposures to heat, noise or carbon monoxides, the proportion of pre-diabetes appeared higher in workers exposed to heat, noise or CO with OR=1.782 (95 %CI: 1.205-2.636), 1.815 (95 %CI: 1.209-2.794) and 1.653 (95 %CI: 1.158-2.361), respectively. Risks for those who were exposed to heat or noise were higher than those who were free from exposure to any occupational hazards ( OR=2.098, 95 %CI: 1.296-3.397). Prevalence rates of pre-diabetes in those who were exposed to heat, noise or CO, were higher than that those who were not. Conclusion:Working hours and exposures to heat, noise or CO appeared as influential factors on PDM.
9.A case-control study on effect of occupational factor exposures on carotid atherosclerosis in steel workers
Zhenwei QIN ; Yajiao SU ; Haitao WANG ; Hongru ZHU ; Xiaotong ZHANG ; Chan LI ; Xiaoming LI ; Zhaoyang WANG ; Juxiang YUAN ; Hongmin FAN
Chinese Journal of Epidemiology 2020;41(11):1836-1842
Objective:To investigate the effect of occupational factor exposures on carotid atherosclerosis (CAS) in steel workers.Methods:A frequency matched case-control study was conducted by age and factory proportion. A total of 1 033 workers with carotid atherosclerosis diagnosed by ultrasonography examination from February to June 2017 were selected as case group, and 1 033 workers without carotid atherosclerosis indicated by physical examination at the same time were selected as control group. The basic information of the workers, such as diet pattern, lifestyle, serum biochemical index and occupation history, were collected. The effects of occupational hazards on carotid atherosclerosis were analyzed by univariate and multivariate logistic regression analyses. The combined effects of various occupational hazards on carotid atherosclerosis were evaluated by environmental risk score (ERS).Results:High temperature, noise, occupational stress and night shift days increased the risk of CAS. With the increase of cumulative high temperature and noise exposure, occupational stress and night shift days, the risk of CAS increased (trend text: χ2=37.53, P<0.01; χ2=16.98, P<0.01; χ2=13.93, P<0.01; χ2=5.59, P<0.05). After adjustment of covariates, compared with P 20 group, the risk of carotid artery in P 40, P 60, P 80 and P 100 groups were as follows: high temperature 1.61 (1.19-2.18), 1.69 (1.25-2.30), 1.84 (1.36-2.49), 2.43 (1.77-3.34); noise 1.70 (1.15-2.52), 1.68 (1.20-2.35), 1.80 (1.34-2.42), 2.23 (1.53-3.26); occupational stress 1.39 (1.04- 1.86), 1.41 (1.06-1.89), 1.45(1.09-1.95), 1.48 (1.10-1.98); night shift days 1.58 (1.08-2.33), 1.66 (1.12-2.47), 1.55 (1.04-2.31), 1.76 (1.17-2.64). The results of the environmental risk score showed that the risk of carotid atherosclerosis increased with the increase of ERS (ERS trend text χ2=51.61, P<0.01); RCS results showed that there was a linear relationship between ERS and CAS in steel workers( P<0.01). Linear dose-response relationship existed between ERS and CAS (nonlinear test P>0.05). Conclusions:High temperature, noise, occupational stress and night shift days were related to carotid atherosclerosis. Linear dose-response relationship existed between ERS and CAS in steel workers.