4.Down-regulation of leucine-rich repeats and immunoglobulin-like domain proteins (LRIG1-3) in HP75 pituitary adenoma cell line.
Dongsheng, GUO ; Lin, HAN ; Kai, SHU ; Jian, CHEN ; Ting, LEI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(1):91-4
Three human leucine-rich repeats and immunoglobulin-like domains (LRIG) genes and proteins, named LRIG1-3, has been previously characterized and it was proposed that they may act as suppressors of tumor growth. The LRIG1 protein can inhibit the growth of tumors of glial cells and the down-regulation of the LRIG1 gene may be involved in the development and progression of the tumor. Real-time reverse transcription-polymerase chain reaction (RT-PCR) is a recently developed technique for quantitative assessment of specific RNA levels. In the current study, it was demonstrated that LRIG1-3 and EGFR mRNA was detected in human pituitary adenoma cell lines and a normal pituitary sample, with differences in the expression levels. Compared to the normal pituitary samples, the expression of LRIG1-3 in HP75 cell line was lower, but the expression of EGFR in HP75 cell line was higher. The results are consistent with LRIG1-3 being tumour suppressor genes, and LRIG genes decreasing the expression of EGFR. The ratio of EGFR/LRIG1 was increased at least 13-fold in HP75 cells compared with the normal pituitary cells, which was also the case for the ratio of EGFR/LRIG2 (14-fold increase in HP75) and EGFR/LRIG3 (11-fold increase in HP75). Further studies were needed to elucidate the explicit role of LRIG genes as negative regulators of oncogenesis in human pituitary adenoma.
5.Construction of a recombinant BCG secreting BP26 and the effects of BP26 on CD4+ and CD8+ T cells in mice
Ting-ting, ZHU ; Lin, ZHANG ; Chuang-fu, CHEN ; Yuan-zhi, WANG ; Jian-xin, LIU ; Hui, WANG
Chinese Journal of Endemiology 2012;31(4):357-360
Objective To develop a BP26 recombinant BCG (rBCG-BP26) vaccine,and to observe the effects of rBCG-BP26 on CD4+,CD8+ T cells in immunized mice.Methods The recombinant shuttle vector pMV261-Ag85B-BP26 was constructed by using traditional molecular biological technology.The recombinant strains were obtained by kanamycin resistance screening and PCR identification after electroporation.Western blotting was used to detect the expression of recombinant BP26 vaccine in immunized mice.Safety experiment was carried out in three different groups:the target experiment(rBCG-BP26) group,the positive control(BCG) group and the negative control(PBS) group,15 BALB/c mice in each group.Intradermal inoculations of 100 μl rBCG-BP26 [containing 106 colony forming units(CFU)],BCG,and PBS were carried out,respectively.Signs of mice in each group were observed.After immunization for 10,20,30,and 40 days,body weight was weighed,and tail blood was collected to observe the change of peripheral blood CD4+ and CD8+ T cells by flow cytometry.Results The rBCG-BP26 was successfully constructed.The expression of BP26 protein was detected in the liquid medium and the bacteria cells.The results of safety test analysis showed that there were no significant differences in signs and body weights(F=2.468,0.331,1.520,0.739,all P> 0.05),between PBS group[ (19.24 ± 0.54),(21.37 ± 0.66),(22.83 ± 0.62),(25.06 ± 0.37)g],BCG group[ (19.90 ± 0.02),(21.53 ± 1.57),(21.95 ± 0.55),(24.70 ± 0.39)g]and rBCG-BP26 group[ (19.16 ± 0.55 ),(20.89 ± 0.20),(22.15 ± 0.76),(24.60 ± 0.64)g].The results of flow cytometry showed that the percentages of CD4+ T cell level were lower in BCG group(26.70%,33.07%) and rBCG-BP26 group( 13.40%,26.70%) than that of the PBS group(33.85%,29.33%) and the values of CD4+/CD8+ T cells increased in rBCG-BP26 group (0.69%,1.27%,1.57%,1.70% ) 10,20 and 30 days after immunization.Conclusions Recombinant BCG-BP26 vaccine strain can express brucella BP26 protein efficiently.Furthermore,its virulence is mild,and it can activate CD4+,CD8+ T cells in the body.It can be used as one of candidate vaccine strain against brucellosis.
6.Prediction of the Secondary Structure and B Cell Epitopes of DMO and DMT Protein in Oreochromis aureus
Jin-Ling CAO ; Jian-Jie CHEN ; Ju-Hua YU ; Ting-Ting WU ;
China Biotechnology 2006;0(07):-
The secondary structure of the protein of DM0 and DMT in Oreochromis aureus were predicted by the methods of Garnier-Robson, Chou-Fasman and Karplus-Schulz based on the amimo acid sequences of DM0 and DMT. And Hydrophilicity plot, Surface probability and Antigenic index for DM0 and DMT protein were obtained by the methods of Kyte-Doolittle, Emini and Jameson-Wolf, respectively. Combined the results according to these methods, the B cell epitopes for DM0 and DMT protein were predicted. The results demonstrated that there were some centers of?-helix in the DM0 protein' s N- terminal No. 80 - 112, 144 -147, 193- 194, 251 - 255, 260 - 269 and No. 279- 283, and in the DMT protein' s N-terminal No. 61 -86, 98 - 105, 140 - 146, 239 -241 and No. 269 -273. And there are some centers of?-sheet in the DM0 protein' s N-terminal No. 59 -61, 69 -70, 148 - 150 and No. 383 -390, and in the DMT protein's N-terminal No. 125 -129, 207-213, 255-264 and No. 281-284. Furthermore, the DMO protein' s N-terminal No. 40-41,44 -45, 50-51, 128-129, 189-192, 204-207, 216-222, 226-233, 244-246, 298 - 299 and No. 323 -326, and the DMT protein' s N-termianl No. 12 - 13, 26 - 27, 43 - 44, 58 - 60, 93 - 95, 115 - 120, 136 -139 and No. 149 -151 may be the flexible regions. Moreover the B-cell epitopes possibly localized in or nearby the DMO protein's 1 -5, 41 -51, 65-67, 86-89, 98-110, 154-170, 183-203, 205 -248, 258-264, 284 - 291, 293 - 298, 270 - 375, 389 - 392 and No. 402 - 410, and DMT protein' s N-termianl No. 1 - 9, 17 - 28, 77 - 84, 114 - 123 , 131 - 139, 157 - 184 and No. 96 - 207. Theses results are helpful for studies on sex control mechanism of DMO and DMT in Oreochromis aureus.
7.Microsurgical treatment of lateral ventricular tumors
Zhengming YANG ; Jian CHEN ; Jincao CHEN ; You ZHOU ; Yuping WANG ; Ping ZHANG ; Wengong BAO ; Ting LEI
Chinese Journal of Microsurgery 2008;31(5):332-334
Objective To investigate the clinical characteristics,minimal invasive operation technique and perioperative management of lateral ventrieular tumors.Methods The clinical characteristics,image diagnosis,surgical approaches and postoperative management and surgical outcomes of 65 consecutive cases of lateral ventricular tumors were retrospectively analyzed.Results In our series,total resection was achieved in 54 cases,and subtotal resection was achieved in 11 cases.Lateral ventricular tumors were mostly found in male and in the left side.Headache caused by increased intracranial pressure was the most common clinical symptom.Ependymocytoma and astrocytic glioma are the most common pathologic diagnosis.Postoperative complications included fever (26 cases),hydrocephalus (9 cases),intraventricular hematoma (7 cases) of which 2 cases were evacuated by craniotomy,epilepsy (7 cases),wound infection (3 cases).Postoperative death was happened in 3 cases.Two of them died of respiratory failure due to postoperative epilepsy.Conclusion Early discovery of lateral ventricle tumors,meticulous operation,subtle micromanipulation and right postoperative treatment are the criticality to improve the rate of total resection of lateral ventricle tumors through microsurgical treatment and reduce postoperative complications and mortality.
8.Microsurgical treatment of tumours in foramen magnum:58 cases report
Liu YANG ; Zhengming YANG ; Wei JIANG ; Jian CHEN ; Jingcao CHEN ; Le DONG ; Ting LEI
Chinese Journal of Microsurgery 2013;36(6):528-531
Objective To explore the optimal diagnosis and treatment strategy of tumors at the foramen magnum,the clinical characteristics were analyzed retrospectively.Methods The clinical data of 58 cases with foramen magnum tumors treated with microsurgery in our department were collected and analyzed retrospectively.And intraoperative neurophysiological monitoring was employed in later 21 cases.Results Gross total resection was achieved in 48 of the 58 cases ; Five of the 10 patients with VA encased by tumors were totally resected.Full resection was achieved in 43 of the last 48 cases.Forty-three of the 47 cases with intradural tumors got complete resection,and total resection was achieved in 3 and 2,respectively,in 9 intra-extradural communicated tumors and 2 extradural ones.Symptoms were significantly improved in 50 cases,worsened in 5 patients,and maintained stable in the last 3 patients.Post-operative symptoms were obviously improved in the later 21 cases with intraoperative neurophysiological monitoring employed and the therapeutic effect of the treatment was much better than the former cases.Three patients died postoperatively.Fifty cases got complete follow-up from 6 months to 3 years.Among them,forty-four patients had a KPS score of 80-100,but the other 6 patients less than 80.Conclusion Microsurgical treatment for foramen magnum tumors is effective.Optimal surgical approaches and precise micro-operative techniques are crucial for the tumor removing.It is quite difficult and risky to fully resect the intra-extradural communicated tumors and the ones encasing vertebral artery.Preoperative evaluation should be profound.The application of intra-operative electrophysiological monitoring and the timely management of post-operative complications can improve the prognosis obviously.
9.HLA-DQA1 genotyping by PCR-SSP technique in Jing nationality of Central Vietnam
Ting-Ping CHEN ; Lin-Lin WANG ; Wei-Xiong LIN ; Jian-Feng CHEN ; Xiang-Zhi XIE ;
Chinese Journal of Immunology 1985;0(03):-
Objective:To investigate the polymorphism of HLA-DQA1 genes in Jing nationality of Central Vietnam.Methods:Applied PCR-SSP tecnique to determine the polymorphism of the HLA-DQA1 alleles of 105 healthy children and youth,unrelated individuals in Central of Vietnam.Results:10 HLA-DQA1 alleles were detected of which DQA1*0104 were the most common allele with frequency of 21.3% and lowest frequency is DQA1*0601.Conclusion:The results indicate that HLA-DQA1 alleles polymorphism of Jing nationality in Central Vietnam is different from the other Chinese. [
10.Influences of ultraviolet A (UVA) on the secretion and expression of chemokine CXCL11/I-TAC by HaCaT cells
Shiling GENG ; Shijun SHAN ; Tongwei ZHANG ; Jian WU ; Zhihua WANG ; Ting XIAO ; Chundi HE ; Hongduo CHEN
Chinese Journal of Dermatology 2010;43(3):181-183
Objective To investigate the influences of UVA on the secretion and expression of chemokine CXCL11/I-TAC by HaCaT cells induced by interferon γ (IFN-γ) and tumor necrosis factor α (TNF-α). Methods HaCaT cells were cultured in the presence of IFN-7 and TNF-a and irradiated with UVA of 2, 4 and 8 J/cm~2, respectively; those cells receiving neither treatment with IFN-γ or TNF-α nor UVA irradiation served as the negative control, and those receiving only cytokine treatment but no irradiation as the positive control. After another 24-hour culture, enzyme-linked immunosorbent assay (ELISA) was performed to detect the protein levels of CXCL11/I-TAC in the supernatant of HaCaT celb, real time PCR to measure the mRNA expression of CXCL11/I-TAC in these HaCaT cells. Results As far as the negative control HaCaT cells were concerned, there was a minor secretion of CXCL11/I-TAC protein and expression of CXCL11/I-TAC mRNA. After treatment with IFN-7 and TNF-a of 10 μg/L, the protein and mRNA expressions of CXCL11/ I-TAC were synergistically upregulated, whereas the induced secretion and expression of CXCL11/I-TAC by HaCaT cells were dose-dependently inhibited by UVA irradiation. Conclusions UVA irradiation inhibits the secretion and expression of CXCL11/I-TAC by HaCaT cells, which in turn suppresses the chemotaxis of Th1/ Tel cells in some degree.