1.Study of mechanism of indirubin derivative PHⅡ-7 in augmenting TRAIL-induced cytotoxicity in breast cancer cell line as well as its chemo-resistant counterpart
Hongwei PENG ; Fei LI ; Xuelian ZHENG ; Yanni LYU ; Xiaochun SUN ; Zhouping DUAN ; Dongsheng XIONG ; Xiaohua WEI
Chinese Pharmacological Bulletin 2015;(5):679-685
Aim To investigate the effect of indirubin derivative PHⅡ-7 and TRAIL on proliferation in breast cancer cell MCF-7 and its MDR counterpart MCF-7/ADR and the mechanism.Methods Growth inhibition rate was examined respectively by MTT assay under treatment with TRAIL or PHⅡ-7 or in combination. Cell apoptosis and ROS production were examined by flow cytometry.The change of TRAIL receptors(DR4/DR5 )in mRNA was analysed by realtime PCR.Re-sults IC50 of PHⅡ-7 on MCF-7 and MCF-7/ADR was (4.49 ±1.55 ),(3.44 ±0.90 )μmol · L-1 respec-tively;MDA-MB-231 was TRAIL sensitive cell line, and apparently TRAIL induced apoptosis in MDA-MB-23 1 .Low concentration of PHⅡ-7 in combination with TRAIL could augment TRAIL-induced cytotoxic effect including apoptosis while TRAIL or PHⅡ-7 treatment alone had limited cytotoxity to those cells.Besides, PHⅡ-7 at this concentration had little toxicity to hu-man peripheral blood mononuclear cells even if in com-bination with TRAIL.PHⅡ-7 generated ROS produc-tion inside MCF-7 and MCF-7/ADR cells and up-regu-lated DR4/DR5 expression concentration dependently. Once upon ROS scavenger NAC involved,the effect of TRAIL receptors up-regualtion by expression was abro-gated.Conclusions PHⅡ-7 at low concentration could improve the sensitivities of breast cancer cell MCF-7 and MCF-7/ADR to TRAIL,the mechanism of which may be the ability of ROS production by PHⅡ-7 help up-regulated TRAIL receptor DR4,DR5 .Our re-search set a solid foundation for PHⅡ-7 in combination with TRAIL in future clinical application.
2.Protective effects of artesunate on septic lung injury in mice via modulation of heme oxygenase-1
Tianhui CAO ; Lei JIANG ; Kai KANG ; Dongsheng FEI ; Shangha PAN ; Songgen JIN ; Mingyan ZHAO
Chinese Journal of Emergency Medicine 2015;24(11):1227-1233
Objective To investigate the effects of artesunate (AS) on septic lung injury in mice and to study the modulation of heme oxygenase-1 (HO-1) in lung in order to clarify the mechanism of AS action.Methods Sixty male Kunming mice were randomly (random number) divided into four groups: Sham group (n =15), CLP group (n =15), AS + CLP group (n =15) and AS + ZnPP + CLP group (n =15).Cecal ligation and puncture (CLP) method was employed to induce septic lung injury.AS (15 mg/ kg) was injected into the abdomen of mice 2 hours before the CLP procedures, and ZnPP Ⅸ, an inhibitor of HO-1, was intraperitoneally injected in dose of 40 μmol/kg 1 hour after the AS injection.The equivalent volume of normal saline was intraperitoneally injected instead in mice of Sham group and CLP group.The mice were sacrificed 24 hours after the CLP procedures.The TNF-α, IL-6 in serum were assayed by ELISA method.The lung injury score and wet/dry ratio were measured.The western blotting and immunohistochemistry methods were used to determine HO-1 protein expression in lung tissue.The protein level of nuclear factor-E2-related factor-2 (Nrf-2), an important transcriptional factor of HO-1 in lung tissue was also analyzed by western blotting.One-way analysis of variance (ANOVA) was used for comparisons among the groups, and SNK-q (Student-Newman-Keuls) test was performed for further comparison, and difference was statistically significant at P < 0.05.Results The TNF-α (pg/mL) (54.37 ± 15.59 vs.627.45 ± 117.03, P < 0.05), IL-6 (pg/mL) (81.53 ± 26.89 vs.898.52 ± 222.78, P < 0.05) in serum was increased, and the lung protein exudation, pulmonary edema (wet/dry weight ratio: 4.27 ± 0.22 vs.6.78 ±0.73, P < 0.05), pulmonary pathology injury (lung injury score: 2.20 ± 0.2 vs.13.25 ± 2.67, P < 0.05) were aggravated by CLP.The HO-1 and Nrf-2 were up-regulated in lung tissue in CLP group compared with the sham group (P < 0.05).After the intervention of AS, the HO-1 and Nrf-2 were further increased (P<0.05), theTNF-α (pg/mL) (627.45 ±117.03 vs.307.88 ±72.33, P<0.05), IL-6 (pg/mL) (898.52 ± 222.78 vs.413.47 ± 115.14, P < 0.05) in serum, lung protein exudation, pulmonary edema (wet/dry weight ratio: 6.78 ± 0.73 vs.5.05 ± 0.61, P < 0.05), pulmonary pathology injury (lung injury score: 13.25 ± 2.67 vs.4.95 ± 1.46, P < 0.05) were attenuated compared with the CLP group.However, the protective role of AS in the septic lung injury in mice was partly reversed by ZnPP, and no significant difference was detected between the AS + CLP + ZnPP and CLP group (lung injury score: 12.15 ± 2.95 vs.13.25 ± 2.67, P > 0.05;wet/dry weight ratio: 6.78 ± 0.73 vs.6.29 ± 0.82, P > 0.05).Conclusions AS plays protective roles in septic lung injury, and it is attributed to limiting lung inflammation via up-regulation of HO-1.
3.Survey of cognitive function in motor neuron disease
Fei GAO ; Dongsheng FAN ; Huali WANG ; Qiong YANG ; Yan ZHANG ; Jun ZHANG ; Yang SHEN
Chinese Journal of Internal Medicine 2009;48(1):31-34
Objective The aim of the study is to investigate the prevalence of cognitive impairment and frontotemporal dysfunction in Chinese patients with motor neuron disease(MND).Methods 100 Datients diagnosed as MND underwent a series of survey including mini mental state examination(MMSE),neuropsychiatry inventory(NPI),Hamilton depression scale(HAMD)and Hamilton anxiety scale (HAMA).Demographics,site of onset,and disease severity-functional rating scale(FRS)were also investigated.Results The prevalence of mild cognitive impairment determined with MMSE score was 24.2%.Compal ison between patients with normal and abnormal MMSE showed statistic differences in depression state and FRS score.Since patients with anxiety and depression would also have abnormal NPI,a follow uP stndy after 3-month antidepressive therapy for the depression patients was made and 2 patients were found to haye Drobably frontotemporal dysfunction lasting for more than 6 months.Conclusion MND patients might have mild impairmented of cognitive function and some of the patients were neuroethologically abnormal.2 of the patients might have frontotcmporal dysfunction.
4.HypnoPTT in the diagnosis of the sleep respiratory disease in children
Dongsheng MIAO ; Lan ZHANG ; Fei HE ; Xuping WANG ; Fenge JIANG ; Yingzhan CHANG ; Weiping LIANG ; Wei LUO
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(05):-
OBJECTIVE To explore a more effective method of detecting sleep respiratory events in children. METHODS Thirty-eight children were tested with HypnoPTT and 31 with polysomnography (control). The test parameters and operative methods were compared. RESULTS In addition to the parameters comm.on to both HypnoPTT and polysomnography, inspiratory flow limitation and spontaneous microarousal can be measured using HypnoPTT and fewer electrodes were needed. CONCLUSION HypnoPTT is a convenient method characterized by less sleep disturbance and credible results, rendering it is especially suitable for pediatric cases. Pulse transit time is a useful parameter for diagnosing the sleep respiratory disease.
5.Fecal diversion in the management of pelvic fractures associated with perineal injuries
Jinlei DONG ; Dongsheng ZHOU ; Lianxin LI ; Yonghui WANG ; Fu WANG ; Maoyuan XIN ; Weifeng WANG ; Fei CHE
Chinese Journal of Orthopaedics 2011;31(11):1209-1212
ObjectiveTo investigate fecal diversion in the management of pelvic fractures associated with perineal injuries.MethodsThis retrospective study involved 27 patients of pelvic fractures associated with perineal injuries during April 2005 to April 2011.There were 23 males and 4 females,and the mean age was 32.9 years old (range,16-62 years old).Fractures type according to Tile classification:4 cases were type A,10 cases were type B,and 13 cases were type C.The pelvic external fixator and fecal diversion were selectively used.Results Of the 27 patients of pelvic fractures associated with perineal injuries,24survived.The overall mortality of pelvic fractures associated with perineal injuries in the present study was 11%.The survived 24 patients were totally reviewed clinically.The mean follow-up time of these patients was 10.9 months (range,4-42 months).Of those patients who underwent early fecal diversion (< 48 h),none experienced infectious completions.However,of those underwent non-early (>48 h) fecal diversion (including those who did not undergo fecal diversion),four patients experienced infectious completions.Fisher's exact test was used to compare the infection rate of these two groups.And the result of Fisher's exact test demonstrated that those patients who underwent early (< 48 h) fecal diversion and non-early (>48 h) fecal diversion (including those who did not undergo fecal diversion).ConclusionStabilization of hemodynamic; selective fecal diversion; early stabilization of pelvic fracture are necessary for the emergency management of pelvic fractures associated with perineal injuries.Rectal injury and severe perineal injury without involvement of rectum should undergo fecal diversion.Early fecal diversion (< 48 h) could reduce the infection rate of pelvic fractures associated with perineal injuries.
6.High volume hemofiltration therapy in patients with multiple organ dysfunction syndrome
Kaijiang YU ; Mingyan ZHAO ; Yan GAO ; Limin LIU ; Hongliang WANG ; Dongsheng FEI
Chinese Journal of General Surgery 2001;0(09):-
Objective To study the effect of high volume hemofiltration(HVHF) in patients with multiple organ dysfunction syndrome (MODS). Methods Nineteen MODS patients were divided into two groups randomly, 10 patients receiving HVHF and 9 patients treated by routine continuous venovenous hemofiltration(CVVH).Artery blood was sampled before and 2、4、8 hours after HVHF and CVVH, concentrations of Scr, BUN, TNF?, IL 1?, IL 6 and blood gas were measured.Results In both HVHF group and CVVH group, the 4th hour Scr、BUN decreased significantly, renal function improved. In HVHF group compared with pre-treatment level the 4th hour concentrations of TNF?[(1 759?506)ng/L vs. (1 265?397)ng/L]、IL-1?[(964?185)ng/L vs. (511?124)ng/L]、IL-6[(1 332?415) ng/l vs. (726?243)ng/L] decreased singificantly. In CVVH group, the 4th hour concentrations of TNF?[(1 799?511) vs.(1 327?421) ng/L] decreased significantly (all P
7.A clinic analysis on 30 cases of mucoepidemoid carcinoma in nasal cavity and nasal sinuses
Yingzhan CHANG ; Wei LUO ; Dongsheng MIAO ; Feng'E JIANG ; Xuping WANG ; Fei HE
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(10):-
OBJECTIVE To evaluate the clinic feature and therapeutic principles and to improve curative effect of mucoepidemoid carcinoma originated in nasal cavity and nasal sinuses. METHODS A restrospective review of clinic features ,therapeutic methods and consequences was undertaken in thirty cases with nasal cavity and nasal sinuse mucoepidemoid carcinoma and treated at the 81st hospital of CPLA from 1990 to 2001. RESULTS Twenty patients received surgery、radiotherapy and chemotherapy, of which sixteen survived in five to ten years follow-up . Six patients received surgery and radiotherapy, of which four survived in five years follow-up. Only one out of two patients who received radiotherapy and chemotherapy respectively survived in five years follow-up. The total five years survival rate was 73 %(22/30). CONCLUSION CT or MRI scan may be helpful to evaluate the location and scale of mucoepidemoid carcinoma.Final diagnosis rely on histopathology examination. Early radical resection combined with radiotherapy and chemotherapy pre- or postoperatively could improve five-year-survival rate and reduce recurrence rate.
8.Effect and mechanism of epidermal growth factor on proliferation of GL15 gliomas cell line.
Heping, WANG ; Dongsheng, GUO ; Fei, YE ; Guifa, XI ; Baofeng, WANG ; Jian, CHEN ; Ting, LEI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(5):604-6
The effects of epidermal growth factor (EGF) on proliferation of G15 glioma cells and the possible mechanisms were investigated. GFAP and EGFR expression was detected by immunohistochemical method. After the cells were treated with EGF at different concentrations, cell count method was used to determine the proliferation of glioma cells, cell cycle and apoptosis were analyzed by flow cytometry (FCM), and laser scan confocal microscope (LSCM) was used to measure the cytoplasmic free calcium. The results showed that GFAP was diffusedly expressed in GL15 cells and EGFR was over-expressed. EGF at doses of < or =1 ng/mL could significantly stimulate cell proliferation, cells in phase G0/G1 decreased, and those in phase S increased. EGF at doses of 10 and 100 ng/ml could inhibit the cell proliferation significantly, and the apoptosis ratio in high dose of EGF group was higher than in control group. EGF could significantly induce a quick rise of intracellular free calcium, but the peak value of intracellular free calcium activated by high dose of EGF was higher than by low dose of EGF. It was suggested that EGF had a dual effect on gliomas: low dose of EGF could stimulate the cell proliferation of gliomas, but high dose of EGF could induce the cell apoptosis and inhibit the proliferation of gliomas, which might be contributed to the difference of intracellular free calcium.
9.Effects of RNAi-mediated gene silencing of LRIG1 on proliferation and invasion of glioma cells.
Feng, MAO ; Baofeng, WANG ; Guifa, XI ; Wei, SUN ; Huaqiu, ZHANG ; Fei, YE ; Dongsheng, GUO ; Ting, LEI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(2):227-32
The effects of RNAi-mediated gene silencing of LRlG1 on proliferation and invasion of the human glioma cell line U251-MG and the possible mechanisms were explored in this study. The plasmids pGenesil2-LRIG1-shRNA1 and pGenesil2-LRIG1-shRNA2 were transfected into U251-MG glioma cells respectively by using Lipofectamine 2000 and the transfected cells in which the LRIG1 expression was stably suppressed were selected by G418. The cells transfected with negative shRNA served as control. The expression levels of LRIG1 mRNA and protein were measured by qRT-PCR and Western blotting, respectively. The cell cycle was analyzed by flow cytometry. The results showed that LRIG1 mRNA expression was reduced by 70% and 58% and LRIG1 protein expression by 58% and 26% in U251-MG cells transfected with pGenesil2-LRIG1-shRNAl and pGenesil2-LRIG1-shRNA2 relative to the negative shRNA-transfected U251-MG cells. The proliferative capacity of the LRIG1 specific siRNA-transfected cells was stronger than that of control cells. Cell cycle analysis showed that silencing LRIG1 significantly increased the percentage of S phase cells and the proliferation index (P<0.01). Moreover, silencing LRIG1 could promote the invasion of U251-MG cells (P<0.05). These findings suggested that LRIG1-targeting siRNA can exert a dramatically inhibitory effect on RNA transcription and protein expression of LRIG1, and LRIG1 down-regulation could promote the proliferation of U251-MG cells, arrest U251-MG cells in S phase, and enhance the invasion of U251-MG cells.
10.Association of expression of Leucine-rich repeats and immunoglobulin-like domains 2 gene with invasiveness of pituitary adenoma.
Huaqiu, ZHANG ; Qin, YAN ; Shutian, XU ; Yibo, OU ; Fei, YE ; Baofeng, WANG ; Ting, LEI ; Dongsheng, GUO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(4):520-3
The Leucine-rich repeats and immunoglobulin-like domains-2 (LRIG2) gene expression in pituitary adenoma and its correlation with tumor invasiveness were studied. The expression of LRIG2 mRNA and protein in human pituitary adenoma obtained surgically was detected by RT-PCR (39 cases) and immunohistochemical staining (30 cases). It was found that LRIG2 was mostly localized at the nucleus of the pituitary adenoma cells. Its expression was significantly higher in the invasive cases than in the non-invasive cases. LRIG2 protein was positive in 14 cases out of 21 cases of invasive adenoma, but only 2 cases were positive in 9 cases of non-invasive adenoma. The positive expression rate of LRIG2 mRNA was 91.3% in invasive cases (total 23 cases) and 62.5% in non-invasive cases (total 16 cases), respectively. LRIG2 gene is overexpressed in invasive pituitary adenoma. It may play an important role in pituitary adenoma invasiveness and further studies are necessary to elucidate the mechanism under this phenomenon.