1.Cell proliferation inhibited by TIP-6 through autophagy in human hepatoma cell line HepG2 and human normal hepatocyte cell line L02
Xiaofei GAO ; Jun HAI ; Yuping DU ; Qin WANG ; Xinping HUI
Journal of Cellular and Molecular Immunology 2009;25(10):883-886
AIM: To investigate the effect of 7-(4-methoxyphenyl)-5, 8a-diphenyl-1,2, 3, 7, 8, 8a-hexahydroimidazo[1,2-a] pyridine (TIP-6) on cell proliferation in human hepatoma cell line HepG2 and human normal hepatocyte cell line L02. METHODS: Typan blue assay was used to check the effect of TIP-6 on cell proliferation. The changes of cell morphology were observed by the phase contrast microscope. Flow cytometry (FCM) was used to check cell cycle. Autophagy and autophagic cell death were detected after acridine orange (AO) staining under fluorescent microscopy. Apoptosis was analyzed by Annexin V/7-AAD, DAPI staining and DNA ladder. NF-κB expression was detected with cellular immunochemistry. RESULTS: Cell proliferation inhibiting effect was appeared when treated with TIP6 from 60 μmol/L to 200 μmol/L, which was correlated with treated concentrations and time. The proliferation rates were just 12.10% and 18.75% (vs control) under 200 μmol/L 72 h in HepG2 and L02 respectively. Vacuolization were found more and more frequently with the increasing of TIP-6 concentrations and treated time prolonged. FCM results indicated that cells were blocked in G2/M phase, and more sensitive were found in HepG2 than L02. AO staining results indicated that the phenomenon of autophagy and autophagic cell death were occurred and appeared more potent with more TIP-6 and longer time treated. No apoptosis markers were found with Annexin V/7-AAD and DAPI staining, and no DNA ladders were found either, these indicated that TIP6 didnt induce apoptosis in these cells. NF-κB was found increased after treated with TIP-6, and the autophagic vacuole became more and more with the increasing of NF-κB protein, but the proliferation rates decreased at the same time. CONCLUSION: TIP-6 inhibited cell proliferation and induced autophagy and autophagic cell death in HepG2 and L02 cells. NF-κB activation may be involved in these effects.
2.Brain White Matter Fiber Change in Major Depression before and after Therapy:a Diffusion Tensor Imaging Study
Hai-Yan LIU ; Zhi-Jian YAO ; Gao-Jun TENG ;
Chinese Mental Health Journal 2002;0(07):-
Objective:To study the cognitive function and brain white matter fiber change in major depressive patients prior and post-treatment.Methods:Eleven major depressed patients were given antidepressants for 10 weeks, and their conditions were evaluated using 24-item Hamilton Depression Scale(HAMD).The cognitive function was determined by using Wisconsin Card Sorting Test(WCST),part of Wechsler memory scale and diffusion tensor ima- ging(DTI)was scanned before and after treatment.11 healthy people as control group were involved and given the same tests at the same time.Results:(1)The WCST scores of patients increased significantly after treatment(prior treatment Cc:1.6?1.6,Re:67.9?20.0,Rpe:51.5?24.8;post treatment Ce:4.0?2.1,Re:43.2?18.8,Rpe:22.8?16.0,P=0.001/0.000/0.003).There was no difference in number sequence memory in Wechsler memory scale.No difference was found between patients after treatment and control group in either WCST or number sequence memory.The patients made significant improvement in the total score of HAMD after treatment(16?14/54?13,t=6.60,P
3.The study of differentiation potential into enteric neuron from rat bone marrow stromul cells in vitro
Yuan-Jun GAO ; Wei QIAN ; Bu-Hai WANG ;
Chinese Journal of Digestion 2001;0(10):-
Objective To explore the differentiation potential of bone marrow stromal cells (BMSC) to enteric neuron in vitro and to seek proper induction methods.Methods BMSC were harves- ted from male rats and cultured in DMEM supplemented with 20% fetal bovine serum,and characterized by flow cytometry.At passage 6,BMSC were pre-induced by basic fibroblast growth factor (bFGF,10 ng/ml) for 24 h,then induced in two groups:glial cell-line derived neurotrophic factor (GDNF) group, 10 ng/ml GDNF in fetal gut condition medium (FGCM) for 10 d.Vitamin A acid (VA) group,VA, zinc in FGCM for 10 d.The expressions of neuronal markers,neural specific enolase (NSE) and neu- rofilament (NF),glial cell marker,glial fibrillary acidic protein (GFAP),enteric neuronal marker,pro- tein gene production 9.5(PGP9.5),nitric oxide synthase (nNOS),enteric neural transmitter,vasoactive intestinal peptide (VIP) were detected by fluorescent immunohistochemistry method.Results The cul- tured BMSC were CD90 positive and CD45 negative on flow cytometry.After 10 d of induction,a certain number of cells adopted neuron-like morphological changes and showed the expressions of NSE,NF, PGPg.5,nNOS and VIP without the expression of GFAP by fluorescent immunohistoehemistry method in both groups.But in GDNF group,the positive rate of NF,PGP9.5,nNOS and VIP was significantly higher than that in VA group (75.6%?8.4% vs 48.5?7.5%;57.7%?6.5% vs 35.7%?7.2% 46.6%?5.4% vs 30.5%?6.6%;72.3%?6.7% vs 40.4%?7.4%;P<0.01).Conclusion BMSC can be induced to differentate into enteric neuron in vitro by different methods.GDNF with FGCM can induce higher rate of enteric neuron like cells compared with VA etc.
4.Immobilized Lipases Cooperates to Catalyze Transesterification Reaction of Lard
Ying HUANG ; Huan GAO ; Hai ZHEN ; Yun-Jun YAN ;
China Biotechnology 2006;0(01):-
The transesterification reaction conditions of lard with methyl acetate with combined use of immobilized lipases as catalysts were conducted. Initially, according to single factorial experiments, the studies on Lipozyme TL IM and Novozym 435 respectively catalyzed transesterification of lard showed that the optimal parameters of transesterification reaction were: the molar ratio of methyl acetate to oil of 14∶1, 40% enzyme added based on oil weight, temperature 50℃. Combined use of Lipozyme TL IM and Novozym 435 was proposed further to improve the catalytic performance by the response surface method (RSM). Herein, a 5-level-3-factor central composite rotated design was employed to evaluate the effects of lipase loading, the proportion of the two lipases and amount of methyl acetate. The optimum conditions were as followings: 40% lipase loading based on oil weight, 50%/50% the proportion of lipases (Novozym 435/Lipozyme TL IM), and the molar ratio of methyl acetate to oil of 14∶1. And under the optimal conditions, the highest biodiesel yield of 97.6% could be attained, which was higher than the biodiesel yield with each single one of the two lipases. The results suggested that the technics of combined use of certain immobilized lipases catalyzed transesterification reaction of lard for biodiesel production with methyl acetate as the acyl acceptor could raise the FAME yield and save the production cost.
6.Evaluation of Urodynamic Study and Somatosensory Evoked Potentials before and after Operation in Children with Lipomyelomeningocele
zhe, GAO ; hai-tao, GU ; gang, WANG ; jun-ping, HE ; yun-fei, GUO ; fang, LIU
Journal of Applied Clinical Pediatrics 2004;0(11):-
Objective To evaluate prourodynamic study and somatosensory evoked potentials(SEPs)before and after operation of children with lipomyelomeningocele(LMMC)and its clinical significance.Methods Urodynamic study(UDS)and SEPs in 31 cases of LMMC who underwent microsurgical release within 1 week preoperatively and 3 months postoperatively were conducted.The 4 parameters used for UDS evaluation were bladder volume,compliance,detrusor activity and residual urine;parameters used for SEPs evaluation were latent period and amplitude.Results There was a statistics significant difference between operation and control group;the similar result was attented in the comparison of before and after operation,also in the comprison of the improved group and the group without improvement after operation.Conclusions UDS and SEPs investigation can provide guidance for the treatment of tethered cord syndrome(TCS)and evaluate clinical prognosis.These cases with the severity changing in UDS and somatosensory shall be avoided unnecessary action in operation.
8.Clinical analysis of 166 aged women with breast cancer
Guo-Xiang GE ; Zhi-Hai GAO ; Xiao-Hui YANG ; Hong-Jun YANG
Chinese Journal of Geriatrics 2000;0(04):-
Objective To evaluate the effect of operation on aged women with breast cancer. Methods The clinical data of the 166 patients recruited from 1989 to 2004 were retrospectively analyzed. Results One hundred and sixty six patients were all treated by operation. The modified radical mastectomy was performed in 98 cases(59.0%), radical masteetomy in 21 cases(12.7%), mastectomy in 29 cases (17.5%), palliative operation in 18 cases (10.8%). After operation, one hundred and twenty six patients received chemotherapy, twelve radiotherapy and 73 endocrine therapy. The rates of postoperative complications were 10.8%, no patients died during operation. The overall postoperative 3-and 5-year survival rates were 82.2% and 65.2% respectively. Conclusions The effect of surgical treatment and its combination with other adjunct therapies is satisfactory for aged women with breast cancer. Different types of operations should be performed for different patients. Ageing is not a contraindication of the operation. Surgery should be combined with medical therapy to reduce the concomitant diseases and postoperative complications.
9.Establishing scientific evaluation view to promote traditional Chinese medicine
Xue-Min GAO ; Jian-Jun ZHANG ; Chun WANG ; Jing-Xia WANG ; Hai-Ying TONG ;
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(11):-
Traditional Chinese Medicine(TCM) makes great contributions to the prosperous growth and people's health.But understanding deviation and imperfect evaluation system of TCM affect the healthy development of TCM.Clinic practice is the motive power of TCM,and curative effect is the key of TCM researches,and the scientific evaluation system is the safeguard for a healthy development of TCM.So we should focus on clinical researches of stubborn diseases and emergency cases to satisfy social demand and upgrade the position of TCM in the medical system.At the same time,functional disease must be explored to show the advantage of TCM.Our mission is to establish a scientific objective evaluation system to accurately understand TCM and take it as the turning point to give an impetus to theoretical breakthrough of the basic studies to promote an overall and healthy development of TCM.
10.Optimization on Fermentation and Purification of the Fibrinolytic Enzyme from Stenotrophomonas maltophilia
Gao-Xue WANG ; Chao-Jun LIANG ; Hai-Hong HUANG ; Jian-Fu WANG ; Ju-Lin YUAN ;
China Biotechnology 2006;0(09):-
The optimization on liquid fermentation and purification of the fibrinolytic enzyme from Stenotrophomonas maltophilia(DR-929) was investigated.The results showed the best fermentation are amidulin 2.0%,soya flour 1.0%,yeast extract 0.5%,NaCl 1.0%,CaCl2 0.02%,MgSO4 0.05%,inoculum of 36 hours,fermental time 4d,initial pH 8.0 or 9.0,temperature 25℃,volume of media 30ml,volume of inoculum 5% or 6%.The purification process includes the following steps: removing cells by the centrifugation,25%~70% saturation ammonium sulfate precipitation,HIC with Phenyl FF(high sub),IEC with Q-Sepharose FF,gel filtration chromatography with Superdex 75.SDS-PAGE electrophoresis was used to examine the purification effect,and the results indicated that homogeneous strap in SDS-PAGE and has a molecular weight around 28.3kDa.The purification factor and activity recovery of the fibrinolytic enzyme are 271.5 and 24.5%,respectively.