1.Inhibition of Bcl-2 Antisense Oligonucleotide Combined with Radiation against Lung Carcinoma Cells
Wei HU ; Guoping SHAN ; Changhui YU
Journal of Zhejiang Chinese Medical University 2007;0(01):-
[Objective] To study the inhibitory effect of bcl-2 antisense oligonucleotide(ASODN)combined with radiation on lung carcinoma cells NCI-H446.[Method]The cultured NCI-H446 lung carcinoma cells were divided into 5 groups:control,pure radiation,nonsense+radiation,lipofectin+radiation,ASODN+radiation.Every group except for control were assayed by MTT on the inhibition rates,24h,48h,72h after 10Gy irradiation respectively.[Results]The inhibitions of lung carcinoma cells NCI-H446 were observed in ASODN+radiation,lipofectin+radiation,nonsense+radiation and pure radiation groups.The differences between the ASODN+radiation group and the other three groups were significant(P
2.Advance in Music Therapy for Speech Rehabilitation (review)
Changhui SUN ; Ruiping HU ; Yulong BAI
Chinese Journal of Rehabilitation Theory and Practice 2013;19(7):623-625
Music and speech may share the same neural pathway, and the role of music therapy in speech rehabilitation gradually attracts the attention from the rehabilitation medical profession in recent years. This article reviewed the effect, mechanism, and strategy of music therapy in speech rehabilitation.
3.A application study of dose constraint template in intensity modulated radiation therapy planning for early stage nasopharyngeal carcinoma
Wei HU ; Changhui YU ; Minghai SHAO ; Haihua YANG ; Weijun DING ; Weifang YANG ; Guoping SHAN ; Shixiu WU
Chinese Journal of Radiological Medicine and Protection 2009;29(1):61-64
Objective To investigate a feasibility of using dose constraint template (DCT) to increase conformity index (CI) of planning target volume (PTV) and improve intensity modulated radiation therapy (IMRT) planning efficiency for early stage nasopharyngeal carcinoma. Methods Ten patients with pathological diagnosed and treated by IMRT were selected for this study. Target volumes were delineated with Corvus 6.3 of treatment planning system, two dose limiting regions(DLR) around PIN were added by extending from PIN,each DLR was 1 cm thick. We created three plans:Plan0,Planl and Plan2. PianO was without DLR and DCT, Planl without DLR but with DCT, Plan2 with both condition;but to compare dose distribution in PLTV and normal tissue using three plans. Results Three plans could fill equal request of dose distribution in PLTV and normal tissue, and their difference was not statistical significant. CI of Plan2 was increased and planning time was decreased significantly compared with Piano and Planl. Conclusloa Usage of DCT together with DLR can increase CI of PTV and improve IMRT planning efficiency for early stage nasopharyngeal carcinoma, planning time is shortened significantly.
4.1,25(OH)2D3 protects against BLM-induced pulmonary fibrosis by inhibiting activation of STAT3
Lishan LUO ; Hangming DONG ; Chaowen HUANG ; Yahui HU ; Changhui YU ; Xuan WAN ; Haijin ZHAO ; Shaoxi CAI
The Journal of Practical Medicine 2017;33(1):59-63
Objective To investigate the impact of 1,25(OH)2D3 on histological changes and activation of STAT3 in BLM?induced pulmonary fibrosis mice. Methods 30 male C57BL/6 mice were randomly divided into control group ,BLM group and BLM+VD group. Mice in BLM group and BLM+VD group received intratracheal injection of BLM(3 U/kg). Control group were intratracheally injected equal volume of sterile saline. From the first day after the surgery,mice in BLM+VD group received intraperitoneal injection of VD (5μg/kg·d). After 21 days, H&E and Masson′s trichrome staining were carried out. Aschroft score were used to evaluate histological changes in lungs. IL?6,IL?4 and INF?γin BALF were assessed by Elisa. p?STAT3,α?SMA and Collagen I were detected by western blot (WB) and immunohistochemistry. Results Fibrosis score and level of α?SMA,Collagen I in BLM group were significantly higher than that in control group (P < 0.05). However ,treatment with VD effectively at?tenuated fibrosis (P<0.05). IL?6 and IL?4 increased while INF?γwas decreased in BALF of BLM group (P<0.05). VD could ameliorate these changes. Upregulation and neuclear translocation of p?STAT3 were observed in BLM group,while VD intervention could inhibit phosphorylation of STAT3. Conclusions VD attenuate BLM?induced pulmonary fibrosis and regulate inflammatory cytokines probably by blocking STAT3 activation.
5.Bcl-2 Antisense Oligodeoxynucleotides Increase Apoptosis of Lung Cancer Cells Induced by Radiation
Wei HU ; Chengchu ZHU ; Min ZHU ; Baoguo CHEN ; Guoping SHAN ; Changhui YU
Journal of Medical Research 2006;0(11):-
Objective To study the effects of Bcl-2 antisense oligodeoxynucleotides(ASODN)on the apoptosis of lung cancer cells induced by radiation in vitro.Methods NCI-H446 lung cancer cell strains were divided into 5 groups:control simple radiation,lipofectin plus radiation,nonsense sqnence radiation and ASODN plus radiation.The cells cultured in five groups were collected at 6h,12h,24h,48h and 72h,with Wright-Giemsa stain,morphology analysis for which was done;the mRNA expression for p53、bcl-2 and PTEN gene was examined by RT-PCR half quantivity and DNA-ploid of the cells in five groups was detected by flow cyfometric method.Results Cell proliferation is obviously restrained and conformation is changed too with the shape crimpled and adherence function decreased obviously after irradiated for 10 Gy dose by the linac;p53 and PTEN expression clearly increased for the combination of Bcl-2 ASODN and bcl-2 mRNA expression clearly decreased.The apoptosis rate after 72 hours among control,pure radiation,lipofectin+radiation,nonsense+radiation and ASODN +radiation grouop is 0.14?0.09,13.17?2.47,11.84?1.76,13.72?1.4,21.26?2.97 respectively,the difference between ASODN combined with radiation grouop and other 4 groups are significant(P
6.Intraoperative lymphatic mapping guided D2 lymphadenectomy in advanced gastric cancer
Jiangwen LIU ; Defeng TONG ; Jianhua NIU ; Junqiang XIA ; Qi WANG ; Changhui DENG ; Jiankun HU
Chinese Journal of General Surgery 2011;26(8):655-658
Objective To compare the number of lymph node dissected by intraoperative lymphatic mapping guided D2 gastrectomy and that by standard D2 gastrectomy plus lymphadenectomy in patients of advanced gastric cancer. Methods In this study 20 advanced gastric cancer cases received intraoperative peritumor injection of carbon nanoparticles suspension ( group 1 ) and D2 lymphadenectomy was guided by the black-stained lymph nodes. 21 cases undergoing standard D2 lymphadenectomy served as controls (group 2). The number of lymph nodes removed and the condition of lymphatic metastasis in two groups, blackstained lymph nodes in group 1, and postoperative complications were compared. Results The average lymph nodes dissected in group 1 (35. 1 ± 13.4) were higher than in control group (26.2 ±7.8). The differences were statistically significant (t =2. 126, P =0. 034). The number of removed N2 and N3 lymph nodes in group 1 were more than that in control group. The total black-stained ration of lymph nodes was 52. 7% in group 1. The positive rate of lymph nodes was higher in black-stained lymph nodes (27.6%) than in unstained lymph nodes ( 10. 8% ) in group 1 and in control group ( 16. 9% ). The differences were also statistically significant ( x2 = 6. 034, P = 0. 016; x2 = 5. 142, P = 0. 023 ). Postoperative afferent loop obstruction developed in one case in group 1. Conclusions Lymphatic mapping guided D2 radical gastrectomy plus lymphadenectomy increases the number of lymph nodes dessected and improves the efficiency of positive lymph nodes excision for patients of advanced gastric cancer.
7.Coculture of human umbilical cord mesenchymal stem cells from Wharton's jelly and brain tumor stem cells
Yi TIAN ; Fangxia GUAN ; Xiang HU ; Bo YANG ; Ying DU ; Changhui ZHOU ; Yuntao BA ; Chenxi GU ; Ningjing LEI ; Xiaowei WANG
Chinese Journal of Tissue Engineering Research 2010;14(10):1721-1728
BACKGROUND:Human mesenchymal stem cells derived from Wharton's jelly(WJCs)display the characteristics of MSCs as defined by the International Society for Cellular Therapy.They can be differentiated into bone,cartilage,adipose,muscle,and neural cells.They can also support the expansion of other stem cells,be weli-tolerated by the immune system,and have the ability to home to tumors.OBJECTIVE:To investigate biological changes of WJCs and brain tumor stem cells(BTSCs)co-cultured in vitro.METHODS:WJCs cultured by situ cultivation and BTSCs used enzyme digestion way respectively,and gathering the 3rd passage of WJCs though subculturing as well as BTSCs.Two kinds of cells co-cultured in 24-well plates in serum-free medium (SFM)without any growth factor.3 and 7 days after co-cultured respectively,CD133 expression of suspension cells in the 24-well plates were identified by flow cytometry,and immunofluorescence was performed for Nestin and glial fibrillary acidic protein (GFAP)expression of adherent cells.Co-culture supernatant(CCS)re-suspended 3~(rd) passage of BTSCs and cultured into 96-well plates at day 3,which were used to determine the difference in cell growth curve in both groups using a microplate reader.RESULTS AND CONCLUSION:With the cocultivation days increasing,the phenomenon that tumor sphere cells began to be decomposed,adherent and differentiated observed by an inverted microscope.BTSCs in the co-cultured group expressed GFAP and Nestin when adherent and differentiated.The higher degree of malignant brain tumor tissue used in culturing BTSCs was,the higher expression of CD133 in BTSCs was.CD 133~+ in BTSCs declined when co-cultured with WJCs.Growth curve of brain tumor stem cells cultured in CCS compared with in SFM at day 3,which indicates that the proliferation of BTSCs inhibited obviously.Results indicated that CD 133~+ expression and proliferative capacity of BTSCs went down and BTSCs underwent differentiation during the co-culture in vitro.
8.Immunomodulatory effect of Wharton's jelly-derived mesenchymal stem cells from human umbilical cord on human peripheral blood T lymphocytes
Changhui ZHOU ; Yi TIAN ; Bo YANG ; Xiang HU ; Hongliang JIAO ; Yunfan ZHOU ; Chengchun WANG ; Chenxi GU ; Ningjing LEI ; Fangxia GUAN
Chinese Journal of Tissue Engineering Research 2010;14(14):2485-2491
BACKGROUND:Bone marrow mesenchymal stem cells have low immunogenicity and immunomodulatory effect,but there are seldom reports concerning the immunomodulatory effect of Wharton's jelly-derived mesenchymal stem cells of human umbilical cord and its mechanims.OBJECTIVE:To investigate the immunomodulatory effects and mechanisms of Wharton's jelly-derived mesenchymal stem cells of human umbilical cord on varient peripheral blood T lymphocytes.METHODS:Mesenchymal stem cells were isolateded from Wharton's jelly of human umbilical cord by tissue culture.T lymphocytes from human peripheral blood were stimulated by phytohemagglutinin and co-cultured with umbilical cord Wharton's jelly-derived mesenchymal stem cells and umbilical cord Wharton's jelly-derived mesenchymal stem cells supernatant respectively to measure A value following 72 hours of coculture using multifunctional microplate reader.Expression of cytokines including transforming growth factor-beta 1(TGF-β1)and interferon-y(IFN-γ)was evaluated by enzyme-labeled immunosorbent assay.RESULTS AND CONCLUSION:Wharton's jelly-derived mesenchymal stem cells could inhibite the proliferation of T lymphocytes induced by phytohemagglutinin.The proliferation inhibition rate was 56%(P<0.01).Wharton's jelly-derived mesenchymal stem cells supernatant also had inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin,in a dose-dependent fashion.The proliferation inhibition rates were 8.3% and 27% respectively in the 50% Wharton's jelly-derived mesenchymal stem cells supernatant and 100% Wharton's jelly-derived mesenchymal stem cells supematant groups(P<0.05).Wharton's jelly-derived mesenchymal stem cells significantly decreased γ-interferon secrted from T-lymphocytes(P<0.05).The secretion of TGF-β1 was lower in the coculture of Wharton's jelly-derived mesenchymal stem cells and T lymphocytes group than Wharton's jelly-derived mesenchymal stem cells alone group(P<0.05).These indicated that Wharton's jelly-derived mesenchymal stem cells and Wharton's jelly-derived mesenchymal stem cells supernatant have inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin.The mechanims may be associated with cell contant and inhibition of v-interferon secrted from T-lymphocytes.
9.High insulin level promotes the degradation of high density lipoprotein generation-related functional protein ABCA1 through calpain and proteasome pathway in 3T3-L1 adipocytes.
Cong YUAN ; Jie WU ; Zhisheng JIANG ; Changhui LIU ; Zewei OUYANG ; Hengjing HU ; Mihua LIU
Chinese Journal of Cardiology 2015;43(2):141-145
OBJECTIVETo explore effects and potential mechanisms of high insulin environment on high density lipoprotein (HDL) generation-related functional protein ABCA1.
METHODS[(3)H] labeled cholesterol efflux from mature 3T3-L1 adipocytes was detected by liquid scintillation counting. ABCA1 mRNA and protein expression in mature 3T3-L1 adipocytes post stimulation with various concentrations of insulin was detected by real-time fluorescence-based quantitative techniques and Western blot, respectively, in the absence and presence of CHX (cycloheximide, CHX), calpeptin (calpain pathway inhibitor) or MG-132 (proteasome pathway inhibitor).
RESULTSCholesterol efflux rates were reduced post insulin stimulation in a dose-dependent manner ((7.06 ± 0.27)%, (6.59 ± 0.30)%, (6.34 ± 0.24)%, (5.07 ± 0.40)%, and (4.71 ± 0.40)% at 0, 1, 10, 10², and 10³ nmol/L of insulin, P < 0.05). Cholesterol efflux rates decreased in a time-dependent manner post 10³ nmol/L insulin stimulation (6.52 ± 0.30)%, (5.59 ± 0.71)%, (5.44 ± 0.37)%, (4.52 ± 0.32)%, and (4.38 ± 0.33)% at 0, 2, 4, 6, 12 h, respectively). ABCA1mRNA levels were not affected by insulin (P > 0.05). ABCA1 protein level was significantly downregulated in 10³ nmol/L insulin group compared to 0 nmol/L insulin group (P < 0.01). Compared with the 0 h group, ABCA1 protein level was significantly reduced in 6 h group (P < 0.05) and further reduced in 12 h group (P < 0.01). Both calpeptin and MG-132 could partly reduce insulin-induced degradation of ABCA1. Compared with the negative control group, ABCA1 protein levels were significantly upregulated by cotreatment with calpeptin and MG-132, respectively (both P < 0.01).
CONCLUSIONOur data suggest that high insulin level could promote the ABCA1 protein degradation and reduce cholesterol efflux from mature 3T3-L1 adipocytes through calpain and proteasome pathway, thus, produce a circumference not suitable for nascent HDL formation in 3T3-L1 adipocytes.
3T3-L1 Cells ; ATP Binding Cassette Transporter 1 ; Adipocytes ; Animals ; Calpain ; Insulin ; Leupeptins ; Lipoproteins, HDL ; Mice ; Proteasome Endopeptidase Complex ; RNA, Messenger
10.Relationship between gene polymorphisms of methylenetetrahydrofolate reductase and plasminogen activator inhibitor-1 and lower limb deep venous thrombosis after colorectal cancer surgery
Zhixiong JIN ; Changhui HU ; Jianjun ZHANG ; Rui LI
Chinese Journal of Postgraduates of Medicine 2023;46(11):972-977
Objective:To analyze the relationship between gene polymorphisms of methylenetetrahydrofolate reductase (MTHFR) and plasminogen activator inhibitor-1 (PAI-1) and lower limb deep venous thrombosis (DVT) after colorectal cancer surgery.Methods:The clinical data of patients with colorectal cancer who underwent surgical treatment in Zhangjiakou First Hospital from February 2020 to February 2023 were analyzed retrospectively. According to the presence or absence of lower limb DVT after surgery, the patients were divided into DVT group (20 cases) and non-DVT group (80 cases). The polymorphism of MTHFR C677T and PAI-1 promoter 4G/5G were detected by polymerase chain reaction method. The relationship between the polymorphisms of MTHFR C677T and PAI-1 promoter 4G/5G and lower limb DVT after colorectal cancer surgery was discussed by logistic regression analysis.Results:TT genotype frequency and T allele frequency of MTHFR C677T in the DVT group were higher than those in the non-DVT group: 65.00% (13/20) vs. 25.00% (20/80), 80.00% (32/40) vs. 38.75% (62/160). CC genotype frequency and C allele frequency were lower than those in the non-DVT group: 5.00% (1/20) vs. 47.50% (38/80), 20.00% (8/40) vs. 61.25% (98/160), with statistically significant differences ( P<0.05). There was no significant difference in CT genotype frequency between the two groups ( P>0.05). 4G/4G gene frequency and 4G allele frequency of PAI-1 gene in the DVT group were higher than those in the non-DVT group: 50.00% (10/20) vs. 21.25% (17/80), 67.50% (27/40) vs. 38.75% (62/160). 5G/5G gene frequency and 5G allele frequency were lower than those in the non-DVT group: 15.00% (3/20) vs. 43.75% (35/80), 32.50% (13/40) vs. 61.25% (98/160), with statistically significant differences ( P<0.05). There was no significant difference in 4G/5G gene frequency between the two groups ( P>0.05). The distribution frequency of TT genotype of MTHFR C677T and 4G/4G genotype of PAI-1 promoter in DVT group was higher than that in non-DVT group: 55.00% (11/20) vs. 22.50% (18/80), with a statistically significant difference ( P<0.05). Multivariate Logistic regression analysis showed that MTHFR C677T TT genotype ( OR = 1.499, 95% CI 1.201 to 1.871), PAI-1 promoter 4G/4G genotype ( OR = 1.471, 95% CI 1.170 to 1.850) and MTHFR The C677T loci TT genotype combined with the 4G/4G genotype of the PAI-1 promoter ( OR = 1.592, 95% CI 1.258 to 2.014) were risk factors for lower limb DVT after colorectal cancer surgery ( P<0.05). Conclusions:The TT genotype of MTHFR C677T site and the 4G/4G genotype of PAI-1 promoter are closely related to the formation of lower limb DVT after colorectal cancer surgery, and the risk of lower limb DVT is higher in patients with both genotype TT and 4G/4G.