1.THE REGWATION OF GANGLIOSIDE ON PKC PATHWAYS AND ITS PROTECTIVE EFFECT ON SERUM-DEPRIVED INJURY IN PC12 CELLS
Ying SONG ; Meng LI ; Jianren LIU ; Sa YU ; Jicheng LI
Acta Anatomica Sinica 1955;0(03):-
Objective To determinted whether GM1 had a protective effect on injury induced by serum-deprivation and the possible mechanism in PC12 cells. Methods The viability of PC12 cells was quantified by MTT after serum-deprivation.The number of apoptotic cells and necrotic cells were determined by Hoechst 33258/PI staining.And the change of PKC protein expression on PC12 cells' membrane and cytosols was detected by Western blotting. Results 1.The viability of PC12 cells decreased after serum-deprivation and the serum-deprivation for 24 hours was chosen as an injury model in this research.Most of the PC12 cells presented apoptosis 24 hours after serum-deprivation.In addition,the PC12 cells' cytosols PKC protein decreased,while the PC12 cells' membrane PKC protein increased significantly,and this result suggested PKC's translocation to membrane and its activation.2.The viability of PC12 cells preincubated with GM1 in high concentrations(10,1,0.1?mol/L) increased significantly and GM1 protected PC12 cells from apoptosis after serum-deprived injury.GM1 reduced the damage of serum-deprivation on PC12 cells and inhibited PKC protein translocation after injury.3.The repair function of GM1 was effective to neuronal resume after serum-deprived injury.Conclusion Neuroprotective effects of GM1 on serum-deprived injury may be partly mediated through the regulation of PKC pathways and it is helpful for the recovery after injury.
2.Controlled live dendritic cell vaccine mediates potent antitumor immune responses
Yu KANG ; Congjian XU ; Xishi LIU ; Xinhua ZHANG ; Chaoqun WU ; Cuiping ZHONG ; Jianren GU ;
Chinese Journal of Obstetrics and Gynecology 2000;0(10):-
Objective To investigate the efficiency of antitumor immune responses induced by a controlled live dendritic cell(DC)vaccine Methods DC precursors were isolated from Fischer 344 rat bone marrow and cultured with granulocyte macrophage colony stimulating factor and interleukin 4 The rat ovarian tumor cell line NuTu 19 was genetically modified by retroviral mediated suicide gene(HSV 1 TK), and the positive clones were selected using G418 Live DC vaccine was then fused with DC and NuTu 19/TK cell by polyethylene glycol The characteristics of live DC vaccine were assayed with flow cytometry and confocal laser scanning microscopy The specific expression of HSV 1 TK gene in live DC vaccine was evaluated by RT PCR and western blot The sensitivity of live DC vaccine to ganciclovir (GCV) was evaluated by methylthiazoletetrazolium assay In vivo, rats vaccinated twice with live DC vaccine were compared to those vaccinated with killed DC vaccine, unfused DC and NuTu 19/TK cell or phosphate buffered saline Seven days following the last immunization, the rats were sacrificed to test the specific cytotoxic T lymphocyte (CTL) activity by lactate dehydrogenase release assay, or challenged with NuTu 19 and tumor incidence was observed Results The fusion efficiency was approximately (23?14) Live DC vaccine displayed an up regulated expression of major histocompatibility complex (MHC) IIOX6 (87 6?3 4)%, costimulatory molecule B 1 2 (71 1?9 3)%, integrin OX 62 (68 0?7 4)%, and adhesion ICAM 1 (77 1?2 0)%, and specifically expressed HSV 1 TK gene. Our data showed that spleen T lymphocytes from rats vaccinated with live vaccine displayed enhanced CTL aetivity (61 8?8 3)% contrast to that of rats vaccinated with killed vaccines (26 0?3 8)% ( P
3.Detection of p53 gene mutations in human leukemia by PCR-SSCP analysis and direct nucleotide sequencing
Rong WANG ; Zhifei YU ; Jianren GU ; Daizhong LI ; Mingchun WANG ; Yuqing CAO
Chinese Medical Journal 1998;111(4):346-350
Objective To look for mutations of the p53 gene in leukimic patients and study the relationship between abnormalities in p53 gene and leukemogenesis.Methods The peripheral blood and Bone Marrow Samples were collected from 36 patients with various leukemia types including 14 cases of lymphocytic leukemia [8 cases of acute lymphocytic leukemia (ALL), 4 cases of chronic lymphocytic leukemia (CLL), 2 cases of hairy cell leukemia (HCL)] and 22 cases of myelocytic leukemia [11 cases of acute non-lymphocytic leukemia (ANLL), 11 cases of chronic myelocytic leukemia (CML)]. DNA structures of exon 5-8 of the p53 gene were scanned by PCR-SSCP (single strand conformation polymorphism analysis of polymerase chain reaction products). The appropriate DNA fragments were amplified, Purified and sequenced directly.Results By PCR-SSCP analysis, shifts in electrophoretic mobility of the p53 gene were detected in 3 of 14 patients with lymphocytic leukemia (2 ALL and 1 CLL), but none in 22 patients with myelocytic leukemia including one in blastic crisis. Direct nucleotide sequencing in one patient with ALL showed transition of CTG to CAG at codon 257 of exon 7, resulting in a change of its encoded amino acids from aspartic acid to valine. To our knowledge, the mutation at this codon has not been previously reported hitherto.Conclusions The p53 gene mutations are specifically associated with lymphocytic leukemia. Alternations of the p53 gene may play a certain role in leukemogenesis in some cases of lymphocytic leukemia.
4.The distribution and structure of class Ⅰ integron in the multidrug-reisistant Pseudomonas aeruginosa
Jianguo CHEN ; Xiaoli DAI ; Yufeng JIANG ; Yingzhao LIU ; Jianren YU ; Zhaoliang SU ; Xinxiang HUANG ; Chiyu ZHANG ; Shengjun WANG ; Qixiang SHAO ; Shihe SHAO ; Huaxi XU
Chinese Journal of Laboratory Medicine 2008;31(8):872-875
Objective To investigate the antimicrobial susceptibility of Pseudomonas aeruginosa (P. aeruginosa) isolated from Zhenjiang area to 13 routinely used antibiotics and identify the structure and dissemination of class Ⅰ integron. Methods K-B test was used to determine the resistant rate of 71 strains of P. aeruginosa. DNA template was extracted by boiling method, PCR method was utilized to detect class Ⅰintegron, and subsequently gene cassettes were analyzed by sequencing. Results The resistant rates to 13 routinely used antibiotics were quite different from 18. 3 to 77.5% among 71 strains of P. aeruginosa. The prevalence of class Ⅰ integron was 38%. These integrons include 5 gene cassettes ( aadB, aac (6) - Ⅱ , PSE-Ⅰ , dfrA17 and aadAS), in which dfrA17 and aadA5 gene cassette were frequently found. Comparing with the negative strains of integron, the positive strains of integron has obviously higher resistance to ten the antibiotics including piporacillin, piperacillin-tazobactam, ceftriaxone, cefepime, ceftazidime, gentamicin,amikacin, tobmmycin, levofloxacin, and ciprofloxacin. Conclusions The resistant rates of P. aeruginosa to 13 drugs were different, and the resistant rates of integron positive strains were obviously higher than integron negative strains, which indicates that integron may play an important role in multidrug reisistance of P. aeruginoosa.
5.Seminoma characterized by thickening of the pituitary stalk:A case report
Bing PENG ; Xingtian WANG ; Yuhuan DENG ; Yu LIAN ; Yanling ZHENG ; Jianren KUANG ; Jinyu QIAN ; Jie LIANG ; Yanlin ZHANG
Journal of Central South University(Medical Sciences) 2024;49(6):863-869
Intracranial seminoma is a rare malignant tumor originating from the germ cells,usually occurring in the pineal gland or pituitary gland.In June 2020,the Department of Endocrinology at the First Affiliated Hospital of Army Military Medical University admitted a 20-year-old male patient with an intracranial germ cell tumor and spinal metastases.The patient presented with headache,dizziness,and visual impairment.Enhanced magnetic resonance imaging(MRI)of the head indicated thickening of the pituitary stalk.After multidisciplinary consultation,the patient underwent endonasal transsphenoidal resection of the tumor,with the pathological diagnosis confirming germ cell tumor.The patient received regular radiotherapy postoperatively.One year later,the tumor recurred and metastasized,leading to a second surgery for tumor resection in the thoracic spinal canal,followed by continued chemotherapy.The patient's clinical symptoms,such as headache and visual disturbances,improved,but he continued to experience panhypopituitarism and required long-term hormone replacement therapy.Early diagnosis of intracranial germ cell tumors is challenging,and they are prone to metastasis and highly sensitive to radiotherapy and chemotherapy.Early diagnosis and multidisciplinary comprehensive treatment can help improve the quality of life and prognosis for patients.