1.Molecular mechanisms of Triple H therapy in the treatment of SAH-induced vasospasm
Ishiguro MASANORI ; B.puryear COREY ; Bisson ERICA ; Jewell RYAN ; J.nathan DAVID ; I.tranmer BRUCE ; C.wellman GEORGE
Journal of Third Military Medical University 2004;26(4):301-303
Objective Cerebral artery vasospasm is a major cause of death and disability in patients experiencing subarachnoid hemorrhage (SAH). Vasospasm typically has been evaluated using angiography to examine narrowing of large diameter (>1 mm) cerebral arteries. Currently, little is known regarding the impact of SAH on small diameter (100~200 μm) cerebral arteries, which play an important role in the autoregulation of cerebral blood flow. The goal of the current study was to examine the influence of SAH on the pressure-diameter relationship of these small diameter blood vessels. Methods Small diameter cerebral arteries were obtained from a rabbit SAH model. Isolated artery segments were canulated and placed in a myograph chamber superfused with warmed, oxygenated, physiological saline solution. Diameter measurements were then recorded to step-wise increases in intravascular pressure. Results Cerebral arteries from SAH animals exhibited a significant increase in pressure-induced constrictions (myogenic tone) at intravascular pressures between 40 mmHg and 120 mmHg. At intravascular pressures above 120 mmHg, myogenic tone began to decrease and was abolished at pressures above 180 mmHg. Conclusion These data suggest that in the days following SAH, myogenic tone is enhanced in small diameter cerebral arteries subjected to physiological levels of intravascular pressure. However, supra-physiological intravascular pressures causes vasodilation, suggesting small diameter cerebral arteries may act as therapeutic targets of hypervolemia, hemodilution, and hypertension "Triple H therapy" used in the treatment of cerebral artery vasospasm.
2.Expression of some apoptosis-related genes in the lung tissues of rats with hypoxic pulmonary hypertension
Guansong WANG ; Guisheng QIAN ; Zhiqiang ZHAO ; Wei YAO
Journal of Third Military Medical University 2001;23(2):140-142
Objective To study the expressions of some apoptosis related to genes in lung tissues and their relationship to the pathogenesis in rats to hypoxic pulmonary hypertension (HPH). Methods A total of 60 rats were employed and equally divided into 5 groups, i.e. control, HPH, hemin, Tin Protoporphyrin (Snpp) and low concentration CO groups. Of them, the rats from middle 3 groups were treated with hypoxia under normal pressure for 7 h in every day, 3 weeks except Sunday. Hemin and Tin Protoporphyrin (Snpp) were given half an hour before hypoxia while low concentration CO after 2 h of hypoxia. The expression of bcl-2, bax, Fas and Fas ligand (FasL) were detected with immunohistochemical staining, in situ cell death detection, DNA fragment detection, and in situ hybridization. Results In low concentration CO and hemin group, typical DNA ladder bands and apoptotic cells were found in lung tissues, and the expressions of Fas, FasL, bax, and bcl-2 mRNA were increased in the epithelial cells of alveoli, vascular and bronchial walls. And the expression of bcl-2 mRNA was decreased than in hypoxic group. Conclusion CO can regulate the expression of bcl-2, bax, Fas and FasL, these apoptotic genes may involve in the regulation of cell apoptosis of lung tissues under HPH and play important roles in the genesis and development of chronic obstructive lung disorders.
3.Telomerase activity and P53 expression in Non-small cell lung cancer and their significances
Tonghua MEI ; Liping ZHANG ; Yuequan JIANG
Journal of Third Military Medical University 2001;23(2):138-139
Objective To investigate the significances of telomerase activity and P53 expression in Non-small cell lung cancer (NSCLC). Methods The activity of telomerase in cancerous tissues was detected by PCR-ELISA, and the expression of P53 protein was detected by S-P method. Results The telomerase activity and P53 expression were significantly higher in NSCLC tissues than in normal lung tissues. The telomerase activity and P53 expression had close association with the differentiation of cancer cells and lymph node metastasis. Conclusion It is very valuable to measure telomerase activity and P53 expression in studying the initiation and progression of NSCLC.
4.Establishment of a multidrug resistance cell line from human lung adenocarcinoma cells and its biologic features
Jie CHEN ; Guisheng QIAN ; Guijun HUANG ; Wei XIONG
Journal of Third Military Medical University 2001;23(2):135-137
Objective To establish a multidrug resistance cell line from human lung adenocarcinoma. Methods The human lung adenocarcinoma cell line SPC-A-1 was exposed to cisplatin of a high and then increasing concentration for 192 d to establish multidrug resistance cell line (SPC-A-1/CDDP). The relative resistance was tested with MTT assay. The morphology of the cells was observed with transmission and scanning electron microscopy and the chromosome of them was analyzed with Giemsa stained specimens. Results The resistance index of SPC-A-1/CDDP cells to cisplatin was 11.2 and the cells showed various cross-resistance to 5-Fu, doxorubicin, mitomycin, vincristine and etoposide, but not to hydroxycamptothecine. Electron microscopy showed the cells with irregular and enlarged nuclei and abundant microvilli. Conclusion A multidrug resistance cell line (SPC-A-1/CDDP) from human lung adenocarcinoma is established. It can be used to downstream experiment.
5.Effect of antisense transfection of monocarboxylate transporter on cell biological characteristics in human lung adenocarcinoma cells
Guizhi ZHANG ; Guijun HUANG ; Xianjian GUO ; Guisheng QIAN
Journal of Third Military Medical University 2001;23(2):128-130
Objective To study the effect of transfecting anti-sense expression vector of the first subtype of the monocarboxylate transporter (MCT1) gene into human lung adenocarcinoma cells on intracellular pH (pHi) regulation, lactate transportation and cell growth. Methods MCT1 antisense gene recombinant vector pLXSN-MCT1 was introduced into human lung cancer cells A549 with electroporation. The cell colonies resistant to G418 were selected. Positive clones were examined by PCR to confirm the integration of genomic A549 DNA and antisene MCT1 gene. The changes of pHi and lactate transportation were detected with spectrophotometry. Cell growth was studied with cell growth curve. Results pHi and lactate transport were remarkably decreased in the transfected cells, and the cell growth was inhibited compared with the cells without transfection(P<0.001). Conclusion MCT1 gene may play an important role in pHi regulation, lactate transport and cell growth in lung tumor cells.
6.Cloning and identification of partial positive regulatory sequence of Na+/H+ exchanger-1 gene from human lung cancer cells
Guoming WU ; Guijun HUANG ; Guisheng QIAN ; Wei YAO
Journal of Third Military Medical University 2001;23(2):125-127
Objective To clone the partial positive regulatory fragment of Na+/H+ exchanger-1 (NHE-1) gene from human lung cancer cells. Methods After BamHⅠ and EcoRⅠ cut sites were added to the 5' ends of the upstream and downstream primers respectively, the partial positive regulatory sequence of NHE-1 gene was cloned with the length of 170 bp from genomic DNA of lung cancer cell line A549 cells with PCR method. The cloned fragment was ligated to plasmid pUC18. Finally, the constructed recombinant was identified with enzyme cut, PCR and DNA sequencing. Results The cloned fragment was about 170 bp in size and successfully ligated to pUC18 with identifiation of double enzyme cut and PCR. DNA sequencing approved that the fragment cloned was objective one with 168 bp in length. Compared with the reported sequence, two t were lost. Conclusion The positive regulatory fragment of NHE-1 gene from human lung cancer cells was successfully cloned.
7.Tumorigenesis of IL-21-transfected HeLa cells in SCID or hu-PBL-SCID mice
Xiaoyun LIU ; Zheng'Ai XIONG ; Haoshan LI ;
Journal of Third Military Medical University 2003;0(08):-
Objective To explore the antitumor effect of interleukin 21(IL-21) after transfected into in HeLa cells in severe combined immunodeficiency(SCID) mice or human peripheral blood lymphocytes-SCID(hu-PBL-SCID) mice.Methods pcDNA4/HisMax-IL-21 was transfected into human cervical cancer cell line HeLa to produce HeLa-IL-21 cells.Totally 48 SCID mice were randomly and equally divided into SCID group(0.2 ml PBS,i.p.) and hu-PBL-SCID group(0.2 ml 2?107/ml hu-PBL,i.p.).Then every group was further divided into 3 subgroups(n=6) to receive a subcutemeous implantation of HeLa,HeLa-vector or HeLa-IL-21 cells,and the rest 6 mice were intraperitoneally injected with anti-asialo GM1 50 mg/animal 1 d before and 4 d after HeLa-IL-21 cells transplantation for NK cell depletion.The growth of the tumor mass was observed.Flow cytometry was used to detect CD8-and DX5-positive cells in hu-PBL from hu-PBL-SCID mice,and spleen NK cells from SCID mice or hu-PBL-SCID mice.The secretion of IFN-? and cytotoxicity of splenic cells from HeLa/HeLa-IL-21-bearing SCID mice or hu-PBL-SCID mice were detected with double sandwich ELISA assay and LDH assay respectively.Results Growth of HeLa-IL-21 tumors was significantly suppressed compared with that of HeLa cells,with a slow development and smaller volume.But this growth suppression was not observed in NK cell depletion groups,with SCID group more severe than hu-PBL-SCID group.hu-PBL-SCID mice bearing HeLa-IL-21 cells(16.55?4.53)had a higher toxicity in spleen cells than those SCID mice bearing HeLa-IL-21 cells(8.32?2.12) or HeLa cells(3.42?1.56)(P
8.Microinjection of adenosine into orexinergic zone of lateral hypothalamus on sleep-wake cycle in rats
Yanping CUN ; Jianxia XIA ; Yang LI ; Zhi'An HU ;
Journal of Third Military Medical University 2003;0(08):-
Objective To investigate the effect of extragenous adenosine(AD) by microinjection in the orexinergic area of rat lateral hypothalamus(LH) on the sleep-wake cycle.Methods A total of 14 male SD rats were given 1,10 or 20 nmol AD and artificial cerebrospinal fluid(aCSF) as own control through three different models of microinjections into the LH.Their electroencephalogram(EEG) and electromyogram(EMG) were recorded by a polygraph recording instrument to observe the changes of sleep-wake after microinjections.Results AD administration to LH at doses of 1,10 and 20 nmol decreased the total amount of wakefulness by 84%,62% and 60% fold during that first 3-hour recording period and concomitantly increased non rapid eye movement(NREM) and REM sleep,when compared with aCSF treatment.The sleep-inducing effect was maintained even in 3 hour after the 20 nmol AD microinjection.The maximal sleep-inducing effect was obtained at the 20 nmol dose of AD administration into the orexinergic LH(P
9.Evaluation of TDP in Treatment of pneumonia in Children
Yongwu WANG ; Yuhua CHEN ; Kaigui CAO ; Huiying DAI ; Defen LUO ;
Journal of Third Military Medical University 1983;0(04):-
110 patients under three years of age with pneumonia were admitted to this hospital in a period from August of 1982 to March of 1983. There were 68 males and 42 females. The patients were randomly divided into Group A and Group B .The symptoms and signs, the laboratory data, and the severity and duration of the illness of the patients of both groups were similar.After admission, laboratory examinations including WBC counts, throat swab cultures and determinations of immunity function were performed and chest x-ray films were taken for all the patients. Regular treatments were given to all of them but the patients of Group A received TDP radiation in addition.After comparing the clinical courses of the two groups, the authors found that TDP could cause more rapid disappearance of cough and moist rales in the lungs, shorten the time of recovery and the whole course of hospitalization, and hasten the absorption of pulmonary infiltrations as seen from the x-ray films. Furthermore, TDP is helpful to promote the immunity function by raising many immune indices. And the rate of lymphocyte transformation was also increased. Its influence on the bringing down of the fever to normal is not very remarkable.It is concluded that TDP radiation is a simple, safe and effective treatment for pneumonia and it is suitable to be used in children.
10.Obervations on TDP Therapy in Treatment of 110 Cases of Infantile Diarrhoea
Xiangying KONG ; Yongwu WANG ; Yuhua CHEN ; Huiying DAI ; Defen LUO ;
Journal of Third Military Medical University 1983;0(04):-
110 cases of infantile diarrhoea were admitted to this institute in a period from Aug. 30 to Dec. 22 of 1982.The patients were randomly divided into two groups, the TDP group and the control group. The general condition and the age distribution of the patients of both groups were similar. The patients of the TDP group received only TDP radiation instead of antibiotics and those of the control group received antibiotics therapy but no TDP. Other treatments such as fluid replacement, dietary regulation, etc, were the same in two groups. Stool samples were sent for routine examination and bacterial culture and blood samples for the determinations of the electrolyte levels, CO2CP, and immunity function for all the patients right after admission as well as just before discharge. The cure rate and course of the disease were similar in two groups. However the pathogenic organisms could still be revealed in the stool of the patients of the TDP group after recovery. But the rate of lymphocyte transformation was significantly higher in the patients of TDP group.It is concluded that TDP radiation is a simple, safe and effective treatment for infantile diarrhoea but its therapeutic mechanism remains obscure.