1.Cytokine detection of tuberculosis patients and study of protective immune mechanism
Jiefan TAN ; Yanya SHEN ; Huijuan FAN
Chinese Journal of Laboratory Medicine 2003;0(07):-
Objective To investigate the roles of the cytokine and chemokine in protective immunity of tuberculosis(TB) patient. Methods The RANTES( CCL5) and MIP1-?( CCL3) content of the Supernatant from PBMC stimulated with IL-4、TNF-?、MIP1-?and LPS in TB patients and normal control subjects was assessed using commercially available ELISA kit. Results The RANTES secretion was increased significantly in both TNF-?plus LPS groups[ (1 731. 86?925. 60) ng/L ] and IFN-?plus LPS [(2 120.78?1 120.72)ng/L] compared with LPS group[ (1 102. 56?873. 44) ng/L] in TB patients (P 0. 05). The MIP1-?secretion in response to IL-4、TNF-?、IFN-?was unchanged significantly in PBMC of TB patients and normal subjects. Conclusion TNF-?、IFN-?and RANTES may play important roles in protective immunity of TB patient
2.The mechanism of inhibitory control in sensation seeking:an event-related potential study
Wenbin SHENG ; Jing XU ; Ya ZHENG ; Fei TAN ; Huijuan SHEN
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(1):25-27
Objective To investigate the neural mechanism of inhibitory control in sensation seeking by using the event-related potential(ERP) technique. Methods High and low sensation seekers( 16 people in each group ), who were selected according to their sensation seeking scores, performed a Go/Nogo task in which the stimuli possessed two levels of difficulty. Electro- encephalogram(EEG) signals were recorded continuously by a set of 32 Ag/AgCI electrodes. Results For two types of stimuli ( Congruent, Incongruent) :( 1 )The amplitude(FCZ) of Nogo N2 and Nogo P3 were( (1.61 ±4.25)μV,(-2.32±4.55)μV)and((16.44±5.74)μV,(17.00±5.71)μV). (2)There was no significant main effects of group for the Nogo N2 amplitude( F (1.30) =0.31, P=0. 59,η2=0. 01;F(1.30) =0.07,P=0.80,η2=0.002) ,the N2d amplitude( F(1.30) =1.18,P=0.29,η2=0.04;F(1.30) =0.004, P=0.95, η2 < 0.001 ) ,the Nogo P3 amplitude( F (1.30) =0.13, P=0.72, η2 =0.004;F(1.30)=0.28, P=0.60, η2 =0.009) and the P3d amplitude( F(1.30) =0.08, P=0.50, η2 =0.02; F (1.30) =0.56,P=0.46, η2 =0.02). (3)Neither of main effects for the N2 and P3 latency was significant (P>0.05). Conclusion The inhibitory control is similar across high and low sensation seeking groups,indicating that there is no relationship between the sensation seeking behaviors and the individual inhibitory control.
3.Experimental study on CT lymphography in evaluating axillary sentinel lymph nodes of rabbit's breast cancer using 3D and curved planar reformation
Pange WANG ; Hongna TAN ; Bo WANG ; Huijuan XIAO ; Jianbo GAO
Journal of Practical Radiology 2017;33(4):615-618
Objective To study the value of CT lymphography (CT-LG) in evaluating axillary sentinel lymph nodes (SLN) of rabbit with breast cancer using 3D and curved planar reformation (CPR).Methods CT-LG examinations were completed on the VX2 rabbit models bearing breast cancer established by tumor tissue mass suspension injection.All raw data were transferred to a postprocessing workstation, then the 3D and CPR were used to show lymphatic vessel and lymph nodes.On the CT-LG images, one or several lymph nodes displayed firstly in the lymphatic drainage pathway was defined as the SLN which was then confirmed by the biopsy.Results (1) 20 rabbits bearing tumor underwnet CT-LG examinations, and both SLNs and draining lymphatic vessels were detected using 3D and CPR techniques.70% (14/20) of experimental rabbits had fine images, and 16 SLNs and 7 non-SLNs were demonstrated on 14 rabbits on CT-LG images with good quality.(2) 8 continuous lymphatic vessels displayed well, exhibiting a homogeneous and continuous tube with uniform contrast-medium filling on CPR images.Most lymphatic vessels demonstrated well, and the 14 vessels with local interrupt or fuzzy on 3D images showed continuous tubular contrast-filling with heterogeneous high-density on the CPR images from the injection sites to axillary SLN.Among 6 rabbits with poor images, 3 rabbits showed the SLN but without lymphatic vessels on 3D images, 2 showed only initial segment of lymphatic vessel,and 1 had no lymphatic vessel and SLN.(3) A total 24 SLNs were obtained by SLNB,including 18 metastatic SLNs and 6 negative ones.On CT-LG images, there were statistical differences between the metastatic and negative SLNs in the maximum diameter and filling defect (P=0.041 and P=0.001, respectively).However, the short diameter, shape and margin of SLNs had no statistical differences between the two groups (all P>0.05).Conclusion On CT-LG examinations, 3D and CPR images may show axillary SLNs and lymphatic vessels effectively,and CPR can demonstrate more details than 3D.The maximum diameter and fillingdefect of SLN are helpful for distinguishing metastatic and negative SLNs.
4.Screening of mouse-derived monoclonal antibodies against the receptor binding domain of Middle East respiratory syndrome coronavirus (MERS-CoV) spike protein
Huijuan WANG ; Wenling WANG ; Jiaming LAN ; Yao DENG ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2016;36(2):88-92
Objective To prepare and screen out monoclonal antibodies against the receptor bind-ing domain (RBD) of Middle East respiratory syndrome coronavirus ( MERS-CoV) spike ( S) protein in mice. Methods The RBD of MERS-CoV S protein expressed in the insect-baculovirus system was purified and then used to immunize the female BALB/ c mice. The spleen cells collected from the mice were fused with myeloma Sp2 / 0 cells. The positive hybridoma cells were obtained by using limited dilution method. Enzyme-linked immunosorbent assay ( ELISA), Western blot assay and neutralization test based on the MERS-CoV pseudovirus were performed for further screening and identification. Results Twelve strains of hybridoma cells that produced the monoclonal antibodies against RBD of MERS-CoV S protein were screened out. All of the 12 monoclonal antibodies (McAbs) could have specific reaction with the RBD of MERS-CoV S protein as indicated by the results of ELISA. Of the 12 McAbs, two were identified as the immunoglobulin M (IgM) isotype and the rest were IgG1 isotype by using double antibodies sandwich ELISA. Four McAbs including 1F1, 2E4, 3C3 and 3E6 were identified as having neutralizing activity by the neutralization test based on MERS-CoV pseudovirus. Results of the Western blot assay showed that the four McAbs (1F1, 2E4, 3C3 and 3E6) could have specific reaction with the RBD of MERS-CoV S protein, but no cross-reac-tion with that of SARS-CoV S protein. Conclusion Twelve mouse-derived McAbs against the RBD of MERS-CoV S protein were obtained. The prepared hybridoma cells showed the characteristics of high speci-ficity and stability in antibody secretion. Four out of the 12 McAbs were proved to have neutralizing activity.
5. Effect of feedback teaching method on rehabilitation training of patients with limb movement disorder after brain glioma surgery
Chinese Journal of Practical Nursing 2018;34(21):1610-1616
Objective:
To explore the application effect of video teaching combined with Teach-back method in rehabilitation training of patients with hemiplegia.
Methods:
The patients with glioma in eloquent area in department of neurosurgery were selected as the research subjects, grouped by time sequence of admission. Between July 2016 to August 2016, 30 patients were selected as control group. Between September 2016 to November 2016, 30 patients were selected as observation group. The control group using conventional department of neurosurgery postoperative routine nursing and hemiplegia rehabilitation education. The observation group was given feedback teaching method on basis of the control group. On the third days after operation, 2 weeks after operation and 4 weeks after operation respectively, the muscle strength, activities of daily living and satisfaction of two groups were evaluated.
Results:
The recovery of muscle strength of the lower limbs of the observation group was better than the control group at 4 weeks after operation (
6.Construction of recombinant infectious clones of HCoV-OC43 expressing green fluorescent protein and viral rescue
Yang YANG ; Liang SHEN ; Roujian LU ; Huijuan WANG ; Wenling WANG ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2016;36(4):256-262
Objective To investigate the feasibility of using recombinant infectious clones of hu-man coronavirus OC43 (HCoV-OC43) as a vector for the expression of exogenous genes and to analyze the insertion sites. Methods Based upon pBAC-OC43FL, a full-length cDNA infectious clone of HCoV-OC43, three recombinant expression plasmids ( pBAC-OC43-GFPΔNS2, pBAC-OC43-GFPΔNS12. 9 and pBAC-OC43-N-GFP) were respectively constructed by replacing NS2 and NS12. 9 genes with the reporter gene en-coding the green fluorescent protein ( GFP ) and inserting the reporter gene after the N gene by using the overlapping-PCR and in vitro ligation. Reverse genetics techniques were used for viral rescue. All of the res-cued virus strains were characterized by immunofluorescence assay ( IFA) and Western blot ( WB) assay af-ter transfecting BHK-21 cells with the recombinant viruses. Results Two recombinant viruses, OC43-GFPΔNS2 and OC43-GFPΔNS12. 9, could be successfully rescued by transfection the BHK-21 cells with pBAC-OC43-GFPΔNS2 and pBAC-OC43-GFPΔNS12. 9 plasmids. The expressed GFP was observed in BHK-21 cells transfected with pBAC-OC43-GFPΔNS2 or pBAC-OC43-GFPΔNS12. 9 plasmids, but not in the cells transfected with the pBAC-OC43-N-GFP plasmid. An efficient and stable expression of GFP was observed in the pBAC-OC43-GFPΔNS2 plasmid-transfected cells. The 10th generation of OC43-GFPΔNS2 virus was ob-tained after repeated freezing and thawing. The expression of GFP and N protein were detected in cells infec-ted with the OC43-GFPΔNS2 virus after 10 passages. Conclusion The NS2 gene of HCoV-OC43 could be used as a promising insertion site of the pBAC-OC43 FL infectious clone for the expression of exogenous genes. This study might provide a platform for further researches on the replication of HCoV-OC43 and the development of human coronavirus-based vectors.
7.The expression of K4 in the tissue of oral submucous fibrosis at the early, middle and advanced stages
Gaoxing WEI ; Yingfang WU ; Aihong TAN ; Xiongwei LIU ; Huijuan ZHAO ; Ersha LIU
Journal of Chinese Physician 2015;17(9):1296-1299
Objective To investigate the expressions of cytokeratin/keratin 4 in the buccal tissue of oral submucous fibrosis at the early,middle and advanced stages,explore which role keratin4 (K4) plays in the process and development of oral submucous fibrosis (OSF),and provide evidence for K4 being a promising biomarker to evaluate the development and prognosis of OSF.Methods Ten cases of normal tissues,and 10 cases of OSF tissues with typical early,middle and advanced stages,were selected,respectively.Detect the expression of K4 in the tissue mentioned above through immunohistochemistry and Westem blot.The data was analyzed by statistical means.Results The results of immunohistochemistry showed that K4 was mainly located in cytoplasm,and positive cells with brownish yellow granules were seen in whole epithelial layer of the normal mucosa.The expression of K4 was lower at all stages of OSF than that at the normal tissue with statistical significance (P <0.05).With the aggravation of OSF,the expression of K4 was decreased,difference between early and advanced stage was found to be statistically significant.The results of Western blot also showed that the expression of K4 was lower than that of early,middle and advanced stages of OSF (P < 0.05).With the aggravation of OSF,the expression of K4 was decreased,but the differences between them had no significance (P > 0.05).Conclusions The expression of K4 in OSF tissue of early,middle and advanced stages were decreased compared to normal tissue,respectively.It suggests that K4 might play and important role in the initiation and development of OSF.
8.Application of reverse dot blotting for detection of Tyzzer’ s organism
Tingting ZHAO ; Lei YAN ; Liwen WEI ; Li WEI ; Huijuan WANG ; Guoqi LAI ; Yi TAN
Acta Laboratorium Animalis Scientia Sinica 2014;(3):72-77
Objective To establish a simple, stable, specific and sensitive method for detection of Tyzzer ’ s or-ganism by reverse dot blotting ( RDB) .Methods Primers and specific probes were designed according to the conservative sequence of Tyzzer 16S rDNA.The forward primer was labeled with biotin .The reverse dot blotting method was established followed by PCR amplification .The specificity and sensitivity of this method were determined .Next, 41 mice and 38 rats were examined by RDB , ELISA and IFA .Results The RDB method showed a high specificity , and in the testing of the 79 laboratory animals , its limit of detection was 4.5 ng/μL.Compared the results of ELISA and IFA , its consistence with ELISA was 100%and the positive rate was 7.59%(6/79), the consistence with IFA was 92.4%(73/79), and the posi-tive rate was 0%.Conclusions An accurate, sensitive and specific method in combination with PCR and RDB in detection of Tyzzer’ s organism is established in this study .
9.Effect of Astragalus Root Injection on hippocampal neuronal apoptosis induced by axaliplatin
Oujing LI ; Huijuan CUI ; Meiyan HUANG ; Huangying TAN ; Yuan LI ; Zhi QI
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(01):-
Objective:To study the effects of Astragalus Root Injection(a traditional Chinese herb medicine) on oxaliplatin-induced apoptosis of hippocampal neurons. Motheds: The primary culture of hippocampus neurons from SD rats (aged within 24h) were treated with different concentrations of oxaliplatin( L-OHP) (0, 3, 5 and 10 ?g/mL) and Astragalus Root Injection (0.05, 0.2 and 0.5 g/mL). The apoptosis of the neurons and the nerve growth factor(NGF)level were analyzed. Results: Astragalus Root Injection dose-dependently decreased neuronal cell apoptosis induced by oxaliplatin (P
10.Factors affecting colorectal cancer and precancerous lesions among cured leprosy patients
Xiaohai ZHANG ; Huijuan SHI ; Hongtao ZHANG ; Hu ZHONG ; Min YAO ; Youji TAN
Journal of Preventive Medicine 2022;34(12):1257-1261
Objective:
To investigate the influencing factors of colorectal cancer and precancerous lesions among cured leprosy patients, so as to provide insights into prevention of colorectal cancer among cured leprosy patients.
Methods:
All cured leprosy patients were recruited from each leprosy village (hospital) in Zhejiang Province from 2019 to 2020. Participants' demographic features, family and medical history of intestinal polyps, smoking, alcohol consumption, dietary behaviors and defecation during the recent three years were collected using questionnaire surveys. Colorectal cancer and precancerous lesions were diagnosed using rectal examination, electronic colonoscopy and histopathological examinations, and factors affecting the development of colorectal cancer and precancerous lesions were identified with a multivariable logistic regression model among cured leprosy patients.
Results:
Totally 100 cured leprosy cases were enrolled, including 72 men and 28 women, and with a mean age of (67.23±3.42) years. There were 13 cases with grade 0 leprosy disability, 16 cases with grade 1 leprosy disability and 71 cases with grade 2 leprosy disability. The detection of colorectal cancer and precancerous lesions was 5.00% and 26.00%, respectively. Multivariable logistic regression analysis showed that age of 65 years and older (OR=7.128, 95%CI: 5.019-10.123), disability (OR=4.591, 95%CI: 2.942-7.163), a medical history of intestinal polyps (OR=4.031, 95%CI: 3.052-5.325), eating moldy food frequently (OR=4.433, 95%CI: 2.920-6.729), seldom eating vegetables (OR=4.411, 95%CI: 3.204-6.071), bloody stools (OR=4.341, 95%CI: 2.771-6.799), mucous stools (OR=4.577, 95%CI: 2.771-7.559) and presence of depression (OR=3.935, 95%CI: 3.026-5.117) were statistically associated with the development of colorectal cancer and precancerous lesions among cured leprosy patients.
Conclusion
Advanced age, disability, history of intestinal polyps, frequent intake of moldy food, infrequent consumption of vegetables, bloody stools, mucous stools, and depression may increase the risk of colorectal cancer and precancerous lesions among cured leprosy patients.