1.Prokaryotic expression of fowl adenovirus serotype 11 Fiber protein and prepara-tion of its polyclonal antibody
Shuaifeng LIU ; Yifan MA ; Xiangqin WANG ; Xiaoran GUO ; Xiaona LIU ; Baishi LEI ; Wanzhe YUAN ; Fuxing CHEN ; Kuan ZHAO
Chinese Journal of Veterinary Science 2024;44(7):1387-1393
In order to obtain polyclonal antibodies against the fibrillar(Fiber)protein of fowl ade-novirus serotype 11(FAdV-11)and investigate its cross-reactivity to different serotypes of FAdV Fiber,the gene encoding the FAdV-11 Fiber protein was cloned into a prokaryotic expression vec-tor pET-32a by homologous recombination technology,then the plasmid was transformed into BL21(DE3)receptor cells,and the purified recombinant protein was used as an immunogen to im-munize rabbits to prepare polyclonal antibody after induced expression,and the cross-reactivity of the polyclonal antibody against different serotypes of FAdV Fiber proteins was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the His-FAdV-11-Fiber recombinant protein was mainly expressed as inclusion bodies and was well expressed.Western blot and IFA showed that the prepared polyclonal antibody reacted with the Fiber proteins of FAdV-8a,FAdV-8b,and FAdV-11,but did not with the 2 Fiber of FAdV-4(Fiber 1 and Fiber 2)proteins.In conclusion,in this study,we successfully prepared rabbit polyclonal antibodies against FAdV-11 Fiber and showed that it specifically recognized the Fiber proteins of FAdV-8a,FAdV-8b and FAdV-11,which lays the foundation for further establishment of serological differential diag-nosis of FAdV-11.
2.Preparation of mouse monoclonal antibodies against the ectodomain of Western equine encephalitis virus E2 (E2ecto) protein.
Fuxing WU ; Yangchao DONG ; Jian ZHANG ; Pan XUE ; Ruodong YUAN ; Yang CHEN ; Hang YUAN ; Baoli LI ; Yingfeng LEI
Chinese Journal of Cellular and Molecular Immunology 2024;40(1):62-68
Objective To prepare mouse monoclonal antibodies against the ectodomain of E2 (E2ecto) glycoprotein of Western equine encephalitis virus (WEEV). Methods A prokaryotic expression plasmid pET-28a-WEEV E2ecto was constructed and transformed into BL21 (DE3) competent cells. E2ecto protein was expressed by IPTG induction and presented mainly as inclusion bodies. Then the purified E2ecto protein was prepared by denaturation, renaturation and ultrafiltration. BALB/c mice were immunized with the formulated E2ecto protein using QuickAntibody-Mouse5W as an adjuvant via intramuscular route, boosted once at an interval of 21 days. At 35 days post-immunization, mice with antibody titer above 1×104 were inoculated with E2ecto intraperitoneally, and spleen cells were fused with SP2/0 cells three days later. Hybridoma cells secreting specific monoclonal antibodies were screened by the limited dilution method, and ascites were prepared after intraperitoneal inoculation of hybridoma cells. The subtypes and titers of the antibodies in ascites were assayed by ELISA. The biological activity of the mAb was identified by immunofluorescence assay(IFA) on BHK-21 cells which were transfected with eukaryotic expression plasmid pCAGGS-WEEV-CE3E2E1. The specificity of the antibodies were evaluated with E2ecto proteins from EEEV and VEEV. Results Purified WEEV E2ecto protein was successfully expressed and obtained. Four monoclonal antibodies, 3G6G10, 3D7G2, 3B9E8 and 3D5B7, were prepared, and their subtypes were IgG2c(κ), IgM(κ), IgM(κ) and IgG1(κ), respectively. The titers of ascites antibodies 3G6G10, 3B9E8 and 3D7G2 were 105, and 3D5B7 reached 107. None of the four antibody strains cross-reacted with other encephalitis alphavirus such as VEEV and EEEV. Conclusion Four strains of mouse mAb specifically binding WEEV E2ecto are successfully prepared.
Horses
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Animals
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Mice
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Encephalitis Virus, Western Equine
;
Ascites
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Immunosuppressive Agents
;
Antibodies, Monoclonal
;
Immunoglobulin M
3.Analysis of tuberculosis epidemiological characteristics and drug resistance among the floating population in Beijing in 2019.
Xin Yu YANG ; Shuang Shuang CHEN ; Jun Li YI ; Yan Feng ZHAO ; Hao CHEN ; Xiao Wei DAI ; Bei Chuan DING ; Meng Di PANG ; Qiao LI ; Zhegn Ying ZHAO ; Chuan You LI
Chinese Journal of Epidemiology 2023;44(6):949-953
Objective: To analyze the epidemic characteristics and drug resistance of pulmonary tuberculosis among the floating population in Beijing and to provide a scientific basis for formulating strategies for the prevention and control of tuberculosis among the floating population. Methods: Data of tuberculosis patients who were positive for Mycobacterium tuberculosis culture was collected from 16 districts and one municipal institution of tuberculosis control and prevention in Beijing in 2019. The strain samples were tested for drug sensitivity by the proportional method. According to household registration location, patients were divided into the floating population and Beijing registration. SPSS 19.0 software analyzed tuberculosis patients' epidemic characteristics and drug resistance in the floating population. Results: In 2019, there were 1 171 culture-positive tuberculosis patients in Beijing, among the floating population, 593 (50.64%) patients were identified, with a male-to-female sex ratio of 2.2∶1 (409∶184). Compared to patients under household registration as Beijing residents, a higher proportion of young adults aged 20-39 years (65.09%,386/593) were noticed, with 55.65% (330/593) reported from the urban areas and 96.80% (574/593) were reported the first time. The differences were statistically significant (all P<0.05). After completing the drug sensitivity test, 37 cases were with multiple drug-resistant tuberculosis, accounting for 6.24% (37/593). The rates of isoniazid resistance (42.11%,8/19) and multidrug resistance (21.05%,4/19) in floating population patients after retreatment were significantly higher than those in newly treated patients (11.67%, 67/574 and 5.75%, 33/574), and the differences were statistically significant (all P<0.05). Conclusions: Most patients with tuberculosis in the floating population in Beijing in 2019 were young males aged 20-39 years. The reporting areas were urban areas and the newly treated patients mainly. The patients with tuberculosis in the re-treated floating population were more likely to suffer from multidrug and drug resistance, which should be taken as the key population for prevention and control.
Young Adult
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Humans
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Female
;
Male
;
Beijing/epidemiology*
;
Tuberculosis
;
Tuberculosis, Pulmonary/epidemiology*
;
Tuberculosis, Multidrug-Resistant/epidemiology*
;
Drug Resistance
4.The Influence of Diabetes, Hypertension, and Hyperlipidemia on the Onset of Age-Related Macular Degeneration in North China: The Kailuan Eye Study.
Yong Peng ZHANG ; Ya Xing WANG ; Jin Qiong ZHOU ; Qian WANG ; Yan Ni YAN ; Xuan YANG ; Jing Yan YANG ; Wen Jia ZHOU ; Ping WANG ; Chang SHEN ; Ming YANG ; Ya Nan LUAN ; Jin Yuan WANG ; Shou Ling WU ; Shuo Hua CHEN ; Hai Wei WANG ; Li Jian FANG ; Qian Qian WAN ; Jing Yuan ZHU ; Zi Han NIE ; Yu Ning CHEN ; Ying XIE ; J B JONAS ; Wen Bin WEI
Biomedical and Environmental Sciences 2022;35(7):613-621
Objective:
To analyze the prevalence of dry and wet age-related macular degeneration (AMD) in patients with diabetes, hypertension and hyperlipidemia, and to analyze the risk factors for AMD.
Methods:
A population-based cross-sectional epidemiologic study was conducted involving 14,440 individuals. We assessed the prevalence of dry and wet AMD in diabetic and non-diabetic subjects and analyzed the risk factors for AMD.
Results:
The prevalence of wet AMD in diabetic and non-diabetic patients was 0.3% and 0.5%, respectively, and the prevalence of dry AMD was 17% and 16.4%, respectively. The prevalence of wet AMD in healthy, hypertensive, hyperlipidemic, and hypertensive/hyperlipidemic populations was 0.5%, 0.3%, 0.2%, and 0.7%, respectively. The prevalence of dry AMD in healthy, hypertensive, hyperlipidemic, and hypertensive/hyperlipidemic populations was 16.6%, 16.2%, 15.2%, and 17.2%, respectively. Age, sex, body mass index, and use of hypoglycemic drugs or lowering blood pressure drugs were corrected in the risk factor analysis of AMD. Diabetes, diabetes/hypertension, diabetes/hyperlipidemia, and diabetes/hypertension/hyperlipidemia were analyzed. None of the factors analyzed in the current study increased the risk for the onset of AMD.
Conclusion
There was no significant difference in the prevalence of wet and dry AMD among diabetic and non-diabetic subjects. Similarly, there was no significant difference in the prevalence of wet and dry AMD among subjects with hypertension and hyperlipidemia. Diabetes co-existing with hypertension and hyperlipidemia were not shown to be risk factors for the onset of dry AMD.
Cross-Sectional Studies
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Diabetes Mellitus/epidemiology*
;
Humans
;
Hyperlipidemias/epidemiology*
;
Hypertension/epidemiology*
;
Macular Degeneration/etiology*
;
Risk Factors
5.Effect of BRCA1 on proliferation, migration and invasion of non-small cell lung cancer H1650 cells via Wnt/β-catenin pathway
YAO Jie1 ; WEI Yaping1, ; LIU Han1 ; LI Hailian1 ; CHEN Qian2
Chinese Journal of Cancer Biotherapy 2021;28(4):353-358
[Abstract] Objective: To investigate the effect of breast cancer susceptibility gene 1 (BRCA1) on the proliferation, migration and invasion of non-small cell lung cancer (NSCLC) H1650 cells through Wnt/β-catenin pathway. Methods: WB and qPCR were used to detect the mRNA and protein expressions of BRCA1 in NSCLC A549, H1299, H1650 cells and normal lung epithelial BEAS-2B cell. A stable BRCA1 over-expression cell line (LV-BRCA1) was constructed in H1650 cells, and blank control group (NC), negative control group (LV-BRCA1-NC), experimental group (LV-BRCA1) and inhibitor group (LV-BRCA1+XAV-939) were set up. The proliferative activity of cells in each group was detected by MTT assay, the migration ability of cells was detected by scratch test, the invasive ability of cells was detected by Transwell method, and the protein expression levels of BRCA1, cyclin D1, β-catenin, c-Myc and Cox2 were detected by WB. Results: The mRNA and protein expression levels of BRCA1 in NSCLC cells were significantly higher than those in BEAS-2B cells (all P<0.01). Up-regulation of BRCA1 expression in H1650 cells could significantly enhance cell proliferation, migration and invasion (P<0.05 or P<0.01), and increase the protein expressions of cyclin D1, β-catenin, c-Myc, Cox2 and c-Jun (P<0.05 or P<0.01). β-catenin inhibitor XAV-939 significantly down-regulated the proliferation, migration and invasion ability of H1650 cells over-expressing BRCA1, and decreased the protein expressions of cyclin D1, β-catenin, c-Myc, Cox2 and c-Jun (P<0.05 or P<0.01). Conclusion: BRCA1 can promote the proliferation, migration and invasion of NSCLC H1650 cells by activating Wnt/β-catenin pathway, and it is expected to be a potential diagnostic biomarker and treatment target for NSCLC.
6.Analysis of mineral contents in the colostrum and peripheral blood and their correlation in pregnant women in Beijing
Yuhui CHEN ; Xiaoyu CHEN ; Yurong ZHAO ; Ping LI ; Kemin QI
Journal of Public Health and Preventive Medicine 2021;32(2):85-88
Objective To investigate the mineral contents of colostrum and peripheral blood and their correlation in pregnant women in Beijing. Methods A total of 80 pregnant women who gave birth in the Affiliated Fuxing Hospital of Beijing Capital Medical University in May 2019 were selected as research subjects, and their colostrum and peripheral blood samples were collected. The contents of 46 minerals in the colostrum and peripheral blood samples were determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and Atomic fluorescence spectrum (AFS). The correlation between the contents of minerals in the colostrum and peripheral blood was analyzed by SPSS 21.0. Results Among the 46 minerals detected, the minerals with higher contents in the peripheral blood were sodium, potassium, iron, magnesium, calcium, zinc, rubidium, copper, aluminum and selenium. The minerals with higher colostrum contents included potassium, sodium, calcium, magnesium, zinc, iron, rubidium, copper, strontium and aluminum. The contents of calcium, strontium, cesium, molybdenum, cobalt, uranium and thorium in the colostrum were significantly higher than those in the peripheral blood (P<0.05). There were significant correlations in the contents of sodium, rubidium, selenium, strontium, cesium, arsenic, mercury and cadmium between the colostrum and peripheral blood (P<0.05). In addition, there were significant correlations between some minerals in the colostrum, for example potassium vs manganese (r=-0.236, P=0.043), rubidium (r=0.816, P<0.001) and magnesium (r=0.400, P<0.001); sodium vs selenium (r =0.509, P<0.001), cerium (r=0.353, P=0.002), praseodymium (r =0.337, P=0.003) and thulium (r=0.331, P=0.004); calcium vs iron (r=0.251, P=0.031) and strontium (r =0.365, P=0.001); magnesium vs cesium (r=0.314, P=0.006) and copper (r =0.395, P=0.001); zinc vs selenium (r=0.310, P=0.007), cerium (r=-0.229, P=0.050), gadolinium (r=-0.372, P=0.001) and yttrium (r=-0.380, P=0.001); rubidium vs titanium (r=-0.413, P<0.001); copper vs cesium (r=0.275, P=0.018); strontium vs uranium (r=0.439, P<0.001); cadmium vs molybdenum (r=0.379, P =0.001). Conclusion The correlations of the mineral contents between the colostrum and peripheral blood suggested that some mineral elements could be accumulated from blood to breast milk through various transport mechanisms. The correlations between the mineral elements in the colostrum showed that there were some interactions between these elements, indicating that maintaining the dynamic balance of mineral elements in human milk is very important for the health of their offspring.
7.Evaluation of shoulder function after repairing head and neck defects with supraclavicular flap.
Jing ZHOU ; Xiao Hong CHEN ; Yong Gang JIN ; Yun Long BAI ; Qi Dong ZHAO ; Yue LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2021;56(9):919-924
Objective: To evaluate the shoulder function in patients after repair of head and neck defects with supraclavicular flap. Methods: A retrospective analysis was performed on 56 patients (54 males, 2 females, aged 35-74 years old) who received the repair of head and neck defects with supraclavicular flaps at Department of Otorhinolaryngology Head and Neck Surgery of Beijing Tongren Hospital, Capital Medical University in January 2013-December 2020. The areas and types of flaps, disruption or infections of the incision at the donor sites and other postoperative complications were recorded. Quick disabilities of the arm, shoulder and hand (Quick-DASH) was used for evaluating the shoulder functions in 43 patients conforming to the standard for evaluation of the clinical functions of shoulders and upper limbs, to compare the postoperative upper limb functions between patients treated with clavicular flaps and patients with acromion flaps. Meanwhile, 30 patients who received bilateral neck lymph node dissection over the same period of time were selected for a comparative evaluation of the donor sides (observation group) and the opposite sides (control group). Data were processed with SPSS 22.0. Results: The areas of obtained supraclavicular flaps were (4-10) cm × (10-18) cm. Three patients (5%) showed the defect widths of 8-10 cm at the donor sites, which couldn't be sutured directly, received the repair of their shoulder defects with partial flaps. Defects in other patients were sutured directly. After surgery, 3 patients (5%) suffered from disruption of the acromion incision, which healed after 2 weeks of local dressing. The follow-up time was 6-43 (27±14) months. All patients expressed no dissatisfaction with the appearance. Among 43 patients, 28 (65%) were clavicular type and 15 (35%) were acromion type. The acromion type showed average motor ability and Quick-DASH scores higher than the clavicular type [(average motor ability: (14.4±4.7) vs. (11.8±3.1), t=2.105, P=0.048; Quick-DASH: (16.9±11.6) vs. (12.2±7.1), t=2.284, P=0.033]. Among 30 patients who received bilateral neck lymph node dissection over the same period of time, the observation group showed higher average motor ability, local symptoms and Quick-DASH scores than the control group [average motor ability: (13.4±5.8) vs. (9.8±4.2), t=3.024, P=0.004; average local symptoms: (4.1±1.0) vs. (3.4±1.0), t=2.537, P=0.014; Quick-DASH: (15.6±14.7) vs. (5.2±11.1), t=3.106, P=0.003]. Conclusion: Shoulder dysfunction exists after treatment with supraclavicular flap, which is related to the flap type.
Adult
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Aged
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Female
;
Humans
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Male
;
Middle Aged
;
Reconstructive Surgical Procedures
;
Retrospective Studies
;
Shoulder/surgery*
;
Skin Transplantation
;
Soft Tissue Injuries/surgery*
;
Treatment Outcome
8. Conditional Deletion of Foxg1 Alleviates Demyelination and Facilitates Remyelination via the Wnt Signaling Pathway in Cuprizone-Induced Demyelinated Mice
Fuxing DONG ; Dajin LIU ; Feiyu JIANG ; Yaping LIU ; Xiuxiang WU ; Xuebin QU ; Jing LIU ; Yan CHEN ; Ruiqin YAO ; Fuxing DONG ; Hongbin FAN
Neuroscience Bulletin 2021;37(1):15-30
The massive loss of oligodendrocytes caused by various pathological factors is a basic feature of many demyelinating diseases of the central nervous system (CNS). Based on a variety of studies, it is now well established that impairment of oligodendrocyte precursor cells (OPCs) to differentiate and remyelinate axons is a vital event in the failed treatment of demyelinating diseases. Recent evidence suggests that Foxg1 is essential for the proliferation of certain precursors and inhibits premature neurogenesis during brain development. To date, very little attention has been paid to the role of Foxg1 in the proliferation and differentiation of OPCs in demyelinating diseases of the CNS. Here, for the first time, we examined the effects of Foxg1 on demyelination and remyelination in the brain using a cuprizone (CPZ)-induced mouse model. In this work, 7-week-old Foxg1 conditional knockout and wild-type (WT) mice were fed a diet containing 0.2% CPZ w/w for 5 weeks, after which CPZ was withdrawn to enable remyelination. Our results demonstrated that, compared with WT mice, Foxg1-knockout mice exhibited not only alleviated demyelination but also accelerated remyelination of the demyelinated corpus callosum. Furthermore, we found that Foxg1 knockout decreased the proliferation of OPCs and accelerated their differentiation into mature oligodendrocytes both in vivo and in vitro. Wnt signaling plays a critical role in development and in a variety of diseases. GSK-3β, a key regulatory kinase in the Wnt pathway, regulates the ability of β-catenin to enter nuclei, where it activates the expression of Wnt target genes. We then used SB216763, a selective inhibitor of GSK-3β activity, to further demonstrate the regulatory mechanism by which Foxg1 affects OPCs in vitro. The results showed that SB216763 clearly inhibited the expression of GSK-3β, which abolished the effect of the proliferation and differentiation of OPCs caused by the knockdown of Foxg1. These results suggest that Foxg1 is involved in the proliferation and differentiation of OPCs through the Wnt signaling pathway. The present experimental results are some of the first to suggest that Foxg1 is a new therapeutic target for the treatment of demyelinating diseases of the CNS.
9.Study on the association between CYP24A1 genetic polymorphisms and risks related to postmenopausal breast cancer
Fuxing CHEN ; Zheng ZHU ; Lan CUI ; Hao YU ; Renqiang HAN ; Pengfei LUO ; Jinyi ZHOU ; Ming WU
Chinese Journal of Epidemiology 2020;41(6):934-939
Objective:To evaluate the associations between CYP24A1 genetic polymorphisms and related risks on breast cancer among postmenopausal women.Methods:We carried out a population-based case-control study to include 1 134 postmenopausal women (589 cases and 545 controls) from Wuxi, Jiangsu province and to explore the association between CYP24A1 polymorphisms and related risks on breast cancer. Seven CYP24A1 variants (rs2209314, rs2585428, rs2762941, rs3787555, rs4909959, rs912505 and rs927650) were genotyped by Sequenom MassARRAY platform. Logistic regression method was used to estimate the CYP24A1 genetic variants and susceptibility of breast cancer. Loci-loci interactions were evaluated by a generalized multifactor dimensionality reduction (GMDR) method.Results:Result showed that rs2209314, rs2585428, rs2762941, rs3787555, rs4909959, rs912505 and rs927650 of CYP24A1 were not associated with breast cancer under the codominant, dominant, recessive or additive models. Among the population with <80 cm waist circumstance, rs2585428 was associated with the reduced risks on breast cancer ( OR=0.64, 95 %CI: 0.42-0.96). Similar negative association was observed for rs3787555 ( OR=0.58, 95 %CI: 0.38-0.87). The genotypes of rs2585428, rs3787555 and rs4909959 showed significant interactions with waist circumstance on the risk of breast cancer. Also, rs2209314, rs3787555 and rs912505 in CYP24A1 could alter the risk of breast cancer by way of loci-loci interaction. Conclusion:CYP24A1 variants rs2585428 and rs3787555 were associated with risks of susceptibility on breast cancer, among postmenopausal women.
10. Status of outpatient service for cancer patients in Beijing Yuetan Community Health Service Center
Chinese Journal of General Practitioners 2019;18(10):977-981
Objective:
To analyze the outpatient service for patients with malignancies in Yuetan Community Health Service Center of Xicheng District in Beijing.
Methods:
Two hundred and nine patients with malignancies visiting Yuetan Community Health Service Center in Beijing from January 2016 to January 2018 were included for analysis. There were 92 patients who visited fixed physicians (fixed group) and 117 patients who visited non-fixed physician (non-fixed group), and the influencing factors affecting fixed physician visit were analyzed.
Results:
Of the 209 patients, 65 (31.10%) were males and 144 (68.90%) were females with a mean age of (69.4±10.0) year. There were 114 cases (54.55%) complicated with, hypertension 52 cases (24.88%) complicated with type 2 diabetes and 40 cases (19.24%) complicated with dyslipidemia. The mean number of visit per patients was (23.16±3.86) times; and the time of visits in fixed group was significantly higher than that in non-fixed group [(27.72±2.67)


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