1.Appraisal of multi-drug resistance function of a new gene cloned from human lung cancer cell line SPC-A-1/CDDP and identify its chromosome′s locus
Guijun HUANG ; Guisheng QIAN ; Weizhong LU
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
objective In the former research work, a differential-expressed gene was cloned from multi-drug resistance lung cancer cell line (SPC-A-1/CDDP) with suppression substractive hybridization, and in this study we further analyze the site of this gene on the chromosome, and appraise its function related to multi-drug resistance in lung cancer cells. Methods The cDNA sequence data of the gene was input to computer and analyzed to ascertain its site on human chromosome by screening the gen bank on the www.ncbi.nlm.nih.gov. With DNA recombination technique, the gene was reversedly inserted to the vector pLXSN to get an antisense expression recombinant vector pLXSN-R, which was then transfected into SPC-A-1/CDDP cells with the aid of electroporation technique. And the semi-quantitative RT-PCR technique was used to quantify the mRNA content of gene in the transfected cells. Finally, the chemosensitivity of the transfected cells was tested with MTT method. Results The gene was located on the human chromosome 19q13.3-19q13.4 locus. The antisense gene recombinant vector was successfully constructed and transfected into SPC-A-1/CDDP cells as shown by its inhibitory activity on the mRNA expression of the gene. The drug-resistance indexes of transfected SPC-A-1/CDDP cells for cisplatin, doxorubicin, 5-fluorouracil, vincristin, hydroxycamptothecine and etoposide were obviously decreased. Conclusion The function of this gene is related to multi-drug resistance in human lung cancer cell, and its locus on the human chromosome is at 19q13.3-1913.4.
2.Clinical Evaluation of Measuring the Cord Blood Gas and Lactic Acid Analysis of Newborns in the Umbilical Artery
Yanping LU ; Guisheng SUN ; Xiayun WENG
Chinese Journal of Perinatal Medicine 1998;0(01):-
Objective To study the plasma lactic acid and blood gas and their value in the umbilical artery of the newborns. Methods The umbilical artery blood samples of 62 newborns were obtained immediately after delivery and then blood gas analysis and plasma lactic acid values were measured within ten minutes. Results The mean value of umbilical artery plasma lactic acid was:(4.4?1.7)mmol/L,pH:7.22?0.07,Base excess:(-5.4?2.7)mEq/L. The lactic acid value had a significant negative relationship with pH values (r=-0.86,P
3.Evaluation of the glucose screening retest during pregnancy
Yanping LU ; Guisheng SUN ; Xiayun WENG ; Lun MAO ; Lian LI
Chinese Journal of Obstetrics and Gynecology 2000;0(12):-
Objective To evaluate of the glucose screening retest for gestational diabetes mellitus (GDM) during pregnancy. Methods A retrospective analysis of 714 pregnant women screened for GDM, between December 1,2001, and December 31,2002, was performed.The first glucose challenge test(GCT) was performed in 16~27 week and retested in 28~38 week.Diagnosis of GDM was based on the criteria of Dong.NDDG criteria was also discussed. Results (1) 1-hour glucose value of 50 g GCT ≥7.8 mmol/L was set as abnormal.The first 50 g GCT abnormal rate was 26.6%(190/714),and the retest abnormal rate was 35.2%(225/639).The mean age of pregnant women in 50 g GCT positive group was significantly higher than that in the negative group( P
4.Role of bcl-2 and bax expression in apoptosis of pulmonary artery smooth muscle cells of rats induced by NHE-1 inhibition in vitro
Junyu LU ; Wei YAO ; Guisheng QIAN ; Guoming WU
Chinese Journal of Pathophysiology 1986;0(03):-
AIM:To investigate the role of bcl-2 and bax expression in apoptosis of pulmonary artery smooth muscle cells(PASMC) of rats induced by Na +/H + exchanger isoform-1(NHE-1) inhibition in vitro. METHODS:Intracellular Ca 2+ concentration ([Ca 2+ ]i) was measured using Fura-2/AM. The expression of bcl-2 and bax mRNA in cells were detected by sqRT-PCR, and the expression of Bcl-2 and Bax protein were examined immunohistochemically. RESULTS:The expression of bax mRNA and Bax protein and [Ca 2+ ]i in cells transfected with NHE-1 ribozyme gene increased significantly compared with those of cells transfected with pLXSN and nontransfected control. Meanwhile, the expression of bcl-2 mRNA and Bcl-2 protein in cells transfected with NHE-1 ribozyme gene decreased significantly. CONCLUSION:Apoptosis of pulmonary artery smooth muscle cells induced by inhibiting NHE-1 may have relevance to the increase in [Ca 2+ ]i and bax expression and the decrease in bcl-2 expression.
5.Clinical Study on Oral Administration and External Application of Yanyuxiao on Acute Traumatic Synovitis of Knee Joint
Guisheng LU ; Jie CHEN ; Xiaopeng GAO ; Hongbin LI
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(3):20-22
Objective To observe the clinical efficacy of oral administration and external application of Yanyuxiao on acute traumatic synovitis of knee joint. Methods Totally 86 patients with acute traumatic synovitis of knee joint were randomly divided into treatment group and control group, with 43 cases in each group. The patients in the two groups were treated with conventional western medicine. Patients in the control group orally took diclofenac sodium at the same time, while patients in the treatment group received oral administration and external application of Yanyuxiao Formula for 2 weeks. The scores of pain, swelling and function of knee joint and the levels interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) of joint fluid before and after the treatment were observed. The efficacy and the recurrence rate in three months after treatment were compared. Results The scores of pain, swelling and function of knee joint of the two groups significantly decreased after treatment, and the scores of pain, swelling and function of knee joint of the treatment group in one and two weeks were lower than those of the control group (P<0.05, P<0.01). The levels of synovial fluid IL-6 and TNF-α of the treatment group were lower than those of the control group after two-week treatment (P<0.01); the total effective rate was higher than that of the control group (P<0.05). The recurrence rate of the treatment group was lower than that of the control group after discharged for 3 months (P<0.05). There were no obvious adverse reactions during the treatment period. Conclusion Oral administration and external application of Yanyuxiao Formula has definite efficacy for acute traumatic synovitis of knee joint, with good safety.
6.Curative observation of Huoxue-Zhitong decoction combined with mannitol in the treatment of swelling and pain after operation of tibiofibular fracture
Xiaopeng GAO ; Guisheng LU ; Jie CHEN ; Yupeng LI ; Chaoxia GAO
International Journal of Traditional Chinese Medicine 2015;(11):994-997
Objective To observe the clinical curative effect of Huoxue-Zhitong decoction combined with mannitol in the treatment of swelling and pain after operation of tibiofibular fracture. Methods 86 patients with swelling after the operation of fracture of tibia and fibula were randomly divided into two groups, with 43 cases in each group. The observation group was treated with intravenous rapid infusion of mannitol, the control group was treated with Huoxue-Zhitong decoction combined with intravenous rapid infusion of mannitol. Swelling, pain visual analogue scale (VAS) score of limb of patients at 1 D and 10 d after operation, time of detumescence and acesodyne and the total efficiency of the treatment between the two groups were compared. Results The indices of the two groups mentioned above at 7 d after the operation significantly decreased, the value in the observation group was lower than the control group (1.3 ± 0.7 cm vs. 1.9 ± 0.9 cm, t=3.451; 1.5 ± 0.6 points vs. 1.9 ± 0.7 points, t=2.845; all P<0.01); The detumescence and acesodyne time of the observation group was shorter than those of the control group (8.6 ± 1.7 d vs. 9.8 ± 2.3 d, t=2.751; 5.5 ± 1.4 d vs. 6.7 ± 1.7 d, t=3.573; all P<0.01), the total efficiency in the observation group was superior to the control group, there were statistically significant differences (97.7% vs. 83.7%; χ2=4.961, P=0.026). Conclusions Huoxue-Zhitong decoction combined with mannitol is effective and safe in the treatment of swelling and pain after operation of tibiofibular fracture.
7.Effects of hammerhead ribozyme on NHE-1 activity and proliferation in pulmonary artery smooth muscle cells of rats in vitro
Wei YAO ; Junyu LU ; Guisheng QIAN ; Xiaojing YANG ;
Journal of Third Military Medical University 1988;0(05):-
Objective To explore the effects of hammerhead ribozyme on the expression and activity of NHE 1 and pHi in pulmonary artery smooth muscle cells (PASMCs) of rats and its role in PASMCs proliferation in vitro . Methods According to the secondary structure of NHE 1 mRNA in rats, NHE 1 specific hammerhead ribozyme was designed with the assistance of computer. The recombinant vector of retroviral plasmid pLXSN and hammerhead ribozyme, PRZ, was transfected into the cultured PASMCs. G418 resistant cell clones were screened with 100 ?g/ml G418. Then, the expression of NHE 1 mRNA was detected by RT PCR, intracellular pH(pHi) value and recovery rate of pHi after intracellular acid loading were measured by fluorescent probe BCECF. 22 Na uptake and 3H TdR incorporation were determined, respectively. Results Compared with the cells transfected with pLXSN and non transfected cells, NHE 1 mRNA, pHi value, pHi recovery rate, 22 Na uptake and 3H TdR incorporation decreased significantly in cells transfected with recombinant vector PRZ. No significance was found between the pLXSN transfected group and non transfected group. Conclusion Hammerhead ribozyme can cleave the target NHE 1 mRNA specifically, reduce the expression of NHE 1 mRNA, induce intracellular acidosis and consequently suppress the proliferation of PASMCs.
8.Adenovirus-mediated protein-kinase-GIα suppresses the hypoxia-induced proliferation and phenotype-switching of pulmonary arterial smooth muscle cell
Bin YI ; Junyu LU ; Li BAI ; Guansong WANG ; Guisheng QIAN
Chinese Journal of Internal Medicine 2010;49(5):385-388
Objective To observe the proliferation and phenotype-switching of pulmonary arterial smooth muscle cell (PASMC) induced by hypoxia and interfered by Ad-PKGIα. And to investigate the potential regulative role of PKGIα gene in the molecule mechanism of hypoxia pulmonary vessel remodeling (HPVR). Methods To establish the pure PASMC cultured by tissue-sticking methods. Semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR) and Western blot were used to examine the PKGIα mRNA and protein expression after PASMC were transfected by Ad-PKG. The mRNA and protein expressive change of smooth muscle α actin(SM-α-actin) determined the degree of cell phenotype-switching. The changes of PASMC proliferation were determined by flow cytometry and ~3H-TdR incorporated way. Results Ad-PKGIα could transfect into PASMC and highly express. Hypoxia down-regulated the expression of SM-α-actin protein (44. 25±5.34 in normoxia, 32. 18±4. 19 in 12 h hypoxia condition, 21.90 ±2. 44 in 24 h hypoxia condition, P < 0. 05), that could be blocked by the transfeetion of Ad-PKGIα. Hypoxia could push PASMC mitosis and proliferating(~3H-TdR incorporated way: 7570 ± 371 in normoxia,12 020± 831 in 12 h hypoxia condition,14 924 ± 1491 in 24 h hypoxia condition, P <0. 05), that could be blocked by the transfection of Ad-PKGIα, too. Conclusions The results suggested that PKGIα signaling pathway might play an important role in the molecule mechanism of HPVR. And PKGIα gene might be a target point of gene therapy.
9.Changes of activity of PKC-AchE and HSP70 in tissues of rabbits dying from suspension of anterior limbs
Yijun ZHANG ; Hualan JING ; Honguian JIN ; Guisheng OU ; Tianyu LU ; Hong LIN ; Xinbiao LIAO
Chinese Journal of Forensic Medicine 2010;25(1):27-29,39,后插1
Objective Investigating activity variance of PKC.AchE and HSP70 in different tissues of rabbits caused by dying from suspension of anterior limbs.in order to study how factors above contribute in the mechanism of death.Methods Suspend rabbit by its anterior limbs until it dies.and coilect tissue from its diaphragmatic muscle,intercostal muscle,myocardium,gastrocnemius,cerebrum,brain stem,liver,kidney and lung.In the control groups,collect same tissues of rabbits dying from strangulation,craniocerebral injury and diseases as contrast.Detect activity of PKC,AchE and HSP70 in tissues of those groups mentioned above by SP stain of immunohistochemistry.and statistically analyze result of detection by rank SUm test of independent samples of multi-groups.Results Compared with data from other control groups,there are more obvious expressions in following tissues of group of suspension:PKC in disphragmatic muscle,intercostal muscles and cerebrum(P<0.05);AchE in disphragmatic muscles,intercostal muscles,myocardium,cerebrum,liver and nerve tract ofmuscles(P<0.05);HSP70 in cerebrum,myocardium and lung(P<0.05).Conclusion Through different mechanisms,PKC,AchE and HSP70 could take some certain contributions in death pmcess of suspension by anterior limbs,and the results above could offer some experimental evidences for related studies and medicolegal investigations.
10.Multimodality imaging evaluation of the treatment effect and mechanism of bone marrow mesenchy-mal stem cells transplantation in swine with acute myocardial infarction
Min CAI ; Rui SHEN ; Lei SONG ; Minjie LU ; Shihua ZHAO ; Yue TANG ; Xianmin MENG ; Guisheng FENG ; Zongjin LI ; Zuoxiang HE
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;(6):420-427
Objective To evaluate the effect and mechanism of bone morrow MSCs transplantation in swine with AMI by cell biology and molecular imaging methods including PET/CT, SPECT, and MRI. Methods Twenty?four Chinese mini?swine ( ( 25 ± 5 ) kg ) were randomly divided into 2 groups: MSCs group ( n=12) and control group ( n=12) . Myocardial infarction was induced in swine hearts by occlusion of the LAD. Thirty minutes later, the MSCs group received autologous MSCs transplantation through in?tramyocardial injection into the peri?infarcted areas (2×107,2 ml) and the control group was subjected to cell culture medium in the same way. At the 1st and 4th weeks after MSCs transplantation, myocardial glu?cose metabolism, myocardial perfusion and cardiac function were evaluated in the two groups through PET/CT, SPECT and MRI. The minimum FDG mean signal intensity ( MSI ) , summed MSI, SRS, SRS%, LVEF, ESV, stroke volume ( SV) and cardiac output ( CO) were calculated. On the 4th week, HE and Masson′s Trichrome stains were performed. Mann?Whitney u test and non?parametric Wilcoxon test were used. Results (1) As evaluated by PET in the 1st week, the MSI and summed MSI in MSCs group were less than those in control group ( 22. 10 ± 3. 18 vs 35. 70 ± 3. 02, z=-2. 65; 1 013. 50 ± 29. 37 vs 1 084. 00 ± 21?15, z=-1.97;both P<0.05) . Compared to the minimum MSI and summed MSI in the 1st week, those in MSCs group increased significantly (34.00±4.25, z=-2.81;1 075.50±28.30, z=-2.80;both P<0?01) in the 4th week. SRS and SRS% decreased in the 4th week compared to those in the 1st week (20.20±2.24 vs 23.80±1.58, (29.80±3.31)% vs (35.10±2.34)%;both z=-2.08, both P<0.05). The averaged MSI in left ventricular infarction area (MSI<70) also increased (56.25±3.54 vs 48.14±2.71;z=-2.80, P<0.01). The a?bove?mentioned parameters had no statistically significant differences in the 4th week compared to those in the 1st week in the control group (all P>0.05). (2) In the 1st week, the perfusion variables had no signifi?cant differences between the two groups ( P>0.05) . There was no significant difference in any perfusion vari?ables between the 1st and 4th weeks in the two groups, respectively (P>0.05). (3) As evaluated by MRI, the cardiac functional parameters had no significant differences between the two groups at the 1st week. In the MSCs groups, LVEF increased significantly ((54.41±2.62)% vs (47.54±2.43)%;z=-2.60, P<0.01) and ESV reduced significantly ((22.85±1.91) vs (27.07±1.67) ml;z=-2.70, P<0.01) in the 4th week com?pared to those in the 1st week; SV and cardiac CO in the 4th week also increased significantly ((29.35± 1?84) vs (26.52±1.46) ml, (2.23±0.14) vs (1.96±0.13) L/min;z=-2.09 and -1.99, both P<0?05). In the control group, there were no significant differences in the cardiac functional parameters between the 1st and 4th weeks ( all P>0.05) . Conclusions Four weeks after MSCs transplantation for AMI, cardiac func?tion and myocardial glucose metabolism improved significantly but without significant myocardial perfusion improvement. Therefore, the cardiac function improvement might be associated with increased myocardial glucose metabolism.