1. The regulatory effects of quercetin combined with DDP on Bcl-2 and Bax expressions in transplanted lung adeno-carcinoma in mice and their significance
Tumor 2008;28(5):386-389
Objective: To observe the inhibitory effects of quercetin combined with DDP on the growth of transplanted lung adenocarcinoma LA795 cells in T739 mice and detect the expression levels of Bel-2 and Bax and apoptotic index to explore the action mechanism for the combination therapy. Methods: Thirty-two T739 mice inoculated with LA795 lung adenocarcinoma cells were randomly divided into four groups with 8 mice in each group: control group (A), DDP treatment group (B), quercetin treatment group (C), DDP plus quercetin treatment group (D). The tumor growth was observed on day 16 after drug administration. All mice were sacrificed on day 24 after inoculation. The weight and volume of transplanted tumors were recorded. The expression levels of Bcl-2 and Bax were quantified using immunohistochemical method and image analyzing system. The apoptotic index of transplanted tumors was measured by TdT-mediated dUTP-biotin nick end labeling(TUNEL) method. Results: The growth of transplanted tumors in groups B, C, and D were significantly inhibited. The weight of tumor body in the three groups were markedly lower than that of group A. The tumor inhibitory rates were 38.57%, 26.03%, and 51.58% in groups B, C, and D, respectively. The inhibitory effect of quercetin combined with DDP on tumor growth was enhanced compared with quercetin or DDP alone treatment. The expression of Bcl-2 was significantly higher but Bax was significantly lower in group A compared with group B, C, and D. The difference between the drug therapy groups and control group was significant (P< 0.05 or P<0.01). The apoptotic index was significantly increased in the three drug therapy groups compared with control group and the difference was significant (P<0.01). Conclusion: The combination therapy for quercetin plus DDP significantly suppresses the growth of lung adenocarcinoma cells in mice. The action mechanism may be due to regulation of Bcl-2 and Bax expressions, thereby, inhibiting cell proliferation and inducing apoptosis.
2. The regulatory effect of ginsenoside Rg3 on expression of somatostatin receptor in lung carcinoma allografts in mice and its significance
Tumor 2012;32(8):572-577
Objective: To investigate the inhibitory effects of ginsenoside Rg3 on the growth and metastasis of lung carcinoma allografts in mice, and to explore the possible mechanism. Methods: The mice bearing a metastatic variant of Lewis lung carcinoma were established, and then they were randomized to receive 0.9% sodium chloride solution (as a control), DDP (cisplatin), and ginsenoside Rg3 from the fourth day after transplant, respectively. Until the twenty-fourth day after transplant, the mice were sacrificed. The subcutaneous tumor was dissected, and the lung was removed. The inhibitory rate of tumor growth and the number of metastatic foci on the lung surface were counted. The expressions of SSTR (somatostatin receptor), VEGF (vascular endothelial growth factor) and PCNA (proliferation cell nuclear antigen) in subcutaneous tumor were examined by immunohistochemistry. The apoptosis was detected by TUNEL (terminal transferase-mediated dUTP nick end-labeling) method. Results: The inhibitory rates of tumor growth in DDP-treated group and the ginsenoside Rg3-treated group were 39.20% and 54.86%, respectively (P < 0.01). The numbers of metastatic foci on the lung surface in DDP-treated group and the ginsenoside Rg3-treated group were decreased by 30.25% and 58.57%, respectively (P < 0.05). The expression level of SSTR and the apoptosis index in the ginsenoside Rg3- treated group were higher than those in the control group and the DDP-treated group (P < 0.01), while the expression level of VEGF and the proliferation index of PCNA in the ginsenoside Rg3-treated group were decreased as compared with the control group and the DDP-treated group (P < 0.01, P < 0.05). Conclusion: Ginsenoside Rg3 can inhibit the growth and metastasis of lung carcinoma allografts in mice. The mechanism may be associated with the overexpression of SSTR. Copyright © 2012 by TUMOR.
3.Effect of peritoneal dialysis fluids on the expression of TLR2 and TLR4 on peritoneal mesothelial cells
Jun WU ; Min HE ; Jian ZHANG ; Wenfei HE ; Bin CHENG ;
Chongqing Medicine 2016;(2):156-158,163
Objective To investigate the effect of glucose-based peritoneal dialysis fluids and icodextrin-based peritoneal dial-ysis fluids on the expression of TLR2 and TLR4 on huamn peritoneal mesothelial cells .Methods Human peritoneal mesothelial cell line 5 - 10 generations(HMrSV5) was cultured in DMEM /F12 medium supplemented with 10% (v/v) fetal calf serum (FCS) .Cell viability and cell proliferation were assessed using M TT method .The experiment were divided into 5 different groups :group A (control group) ,1 .5% dextrose group ,2 .5% dextrose group ,4 .25% dextrose group and 7 .5% Lcodextrin group .Icodextrin group (aikau dextrin) ,TLR2 and TLR4 expression were detected by Western blot .Results Treatment with different concentrations of glucose-based peritoneal dialysis fluids for 24 h did not affect the expression of TLR2 and TLR4 protein .In addition ,after stimula-tion for 48 h ,1 .5% dextrose ,2 .5% dextrose ,4 .25% dextrose decreased TLR2 expression by (5 .5 ± 2 .8)% ,(31 .4 ± 7 .5)% , (54 .9 ± 1 .9)% respectively ,TLR4 expression by (32 .9 ± 17 .6)% ,(47 .7 ± 13 .5)% ,(66 .4 ± 13 .5)% respectively .Stimulation for 72 h ,decreased TLR2 expression by (29 .4 ± 14 .7)% ,(38 .9 ± 9 .9)% ,(63 .5 ± 16 .5)% respectively ,TLR4 expression by(59 .5 ± 16 .8)% ,(63 .1 ± 9 .5)% ,(79 .2 ± 14 .0)% respectively .There was no significant change in TLR2 and TLR4 protein expression on 7 .5% icodextrin group .Conclusion Glucose-based peritoneal dialysis fluids ,but not icodextrin-based peritoneal dialysis fluids downregulates expression of TLR2 and TLR4 by HM rSV5 .
5.Application of total hip replacement for intertrochanteric fractures following the failure of dynamic hip screw fixation in four elderly people
Shuangjian HE ; Zheng YE ; Jian ZHU ; Weihua LUO ; Bin YAN
Chinese Journal of Tissue Engineering Research 2010;14(17):3210-3212
BACKGROUND:Many internal fixations,such as dynamic hip screw,Gamma screw,proximal femoral nail,angle steel plate,as well as locking proximac femoral plate,are utilized in treating intertrochanteric fractures,especially the dynamic hip screw However,the failure rate is gradually increased.OBJECTIVE:To explore the application and clinical efficacy of total hip replacement for the treatment of elderly intertrochanteric fracture fixation after failure of dynamic hip screw.METHODS:A total of four cases with intertrochanteric fractures were treated by total hip replacement after failure of dynamic hip screw fixation was selected.According to Evans typing,one case were type Ⅱ,two cases were type Ⅲ A,and one case was typeⅢB.Internal fixation displacement could be found at half to 1 year after dynamic hip screw fixation.Because of bone disunion,coxa adducta and pain,the patients could not walk.Sequentially,total hip replacement was performed with 45°abduct angle and10°-15°anteversion angle The clinical efficacy was evaluated by Harris scoring criteria.RESULTS AND CONCLUSION:All the cases were operated smoothly,with 1.5-2 hour operation duration and 400-600 mL blood loss.No case appeared allergic response to bone cement.By 3-12 months follow-up,4 successful operative cases do not appear prosthesis loosening,subsidence and are satisfied with well hip function.The average Harris score were 81 points.The results revealed that application of total hip replacement after the failure of DHS in elderly intertrochanteric fracture fixation,which shortened the time patients stay in bed to reduce complications and improved the hip joint function.
6.Quantitative measurement of fatty liver by spectral imaging on gemstone CT:an experimental study on mice model
Tingting SHI ; Jian HE ; Jiong SHI ; Shan JIANG ; Bin XUE ; Zhengyang ZHOU ; Bin ZHU
Journal of Practical Radiology 2014;(12):2079-2083
Objective To investigate the feasibility of quantitative measurement of fat concentration by CT spectral imaging in a mice model of fatty liver.Methods Twenty-four mice with different degrees of fatty liver underwent CT spectral imaging.CT values of liver parenchyma under mixed X-ray energy and 65 keV,fat concentration based on various basic material pairs (fat/water,fat/io-dine,fat/calcium)and spectral curves were obtained.Liver specimens were obtained to measure the concentration of triglyceride , and HE staining was performed.Correlations between various CT indexes and triglyceride concentration were analyzed.Results Correlation between fat concentration (fat/water pair)and triglyceride (r =0.91 5 )was better than that between CT values on 65 keV and triglyceride (r=-0.858),as well as polychromatic CT values (r=-0.81 6).All the P values were<0.001.Correlations between fat concentrations based on fat/iodine or fat/calcium pairs and triglyceride were relatively low (r=-0.726,-0.660).CT indexes on 1.25 mm slice thickness performed better than those on 2.5 mm.With fatty liver degree increased,the shape of spectral curve changed gradually.Conclusion Liver fat concentration can be measured by CT spectral imaging noninvasively,accurately and quantitatively in a mice model of fatty liver.
7.Suppression of corneal neovascularization by culture supernatant of human amniotic epithelial cells in vitro
Bin-bin, LI ; Xiao-xi, YANG ; Qing, ZHOU ; Yan-hua, HE ; Jian, CHEN
Chinese Journal of Experimental Ophthalmology 2012;30(5):408-413
BackgroundCorneal neovascularization (CNV) is a common eye disease.The researches on the pathogenesis and treatment of CNV are focus in Ophthalmology.ObjectiveThis study was to investigate the effects of culture supernatant from human amniotic epithelial cells (AECs) on CNV in vitro and its mechanism.MethodsHuman AECs were obtained from a placenta and cultured in serum-free medium for 48 hours,and the supernatant was collected.The levels of interleukin-1 receptor antagonist (IL-1 Ra) and pigment epithelium-derived factor (PEDF) in the human AECs culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA).Rabbit corneal epithelial cells (CECs) were obtained and cultured in different concentrations of human AECs culture supernatant for 48 hours,serum-free medium was used as the control group.The expression of vascular endothelial growth factor (VEGF) mRNA and basic fibroblast growth factor (bFGF) mRNA in rabbit CECs were measured by quantitative real-time PCR (QRT-PCR).Human umbilical cord vein endothelial cells (UVECs) were cultured in the three mediums above,and the proliferation of human UVECs (absorbance value,A value) was tested by the cell counting kit 8 ( CCK8 ).Migration assay was performed by the wound healing method for the human UVECs.The membrane ultra-structure of human UVECs was examined under the atomic force microscope (AFM).ResultsCultured and passaged human AECs showed a positive response for keratin.The expression of VEGF mRNA (1.00±0.22) and bFGF mRNA (1.00±0.36) in rabbit CECs was suppressed by the human AECs culture supernatant,with a significant reduction in comparison with the serum-free DMEM group (2.98±0.46,2.55±0.48 )(P=0.001,0.002).The A value was significantly lowered in the human AECs culture group for 72 hours compared with the serum-free DMEM group ( 1.941 ± 0.036 versus 2.144 ± 0.059 ) ( P =0.000 ),and the bFGF-induced migration rate of human UVECs was strongly inhibited by the culture supernatant of human AECs in comparison with serum-free DMEM.The plasma membrane of human UVECs cultured with the human AECs culture supernatant was full of bumps,and decreased intercellular connection and cellular pseudopodia were found on the AFM image.The concentration of IL-1Ra was (153.56±0.36)ng/L and that of PEDF was (70.41 ±0.68 )μ,g/L in the human AECs culture supernatant.Nothing was deteched in serum-free DMEM group.Conclusions Human AECs culture supernatant suppressed the expression of VEGF and bFGF in CECs as well as the migration and proliferation of vascular endothelial cells.The effect may be associated with IL-1Ra and PEDF secreted by human AECs.These results suggest that human AECs may be a potential therapy for the inhibition of CNV.
8.Imageology and significances of rotational axes of distal femur on Chinese adults with total knee arthroplasty.
Bing-hua SONG ; Jun-ying SUN ; Zeng-liang NI ; Bin HE ; Jian-qun HE ; Rong SHAO
China Journal of Orthopaedics and Traumatology 2016;29(1):41-47
OBJECTIVETo explore clinical significance of rotational axis of distal femur on Chinese adults in total knee arthroplasty (TKA).
METHODSThere were 86 Chinese adults (106 normal knees) including 47 males (53 knees) and 39 females (53 knees), 54 knees were on left and 52 on right. The CT scan was employed in the distal femur. The scan direction was aligned to be on the plane perpendicular to the mechanical axis of the femoral. The CT images of cross sections across lateral and medial femoral epicondyle were moved to personal computer,lateral angle between anterior posterior line (APL) and surgical transepicondylar axis (STEA) (ATA),lateral angle between posterior condylar line (PCL) and APL (APA), angle between perpendicularity of APL and PCL (A-PA), posterior condylar angle (PCA), condylar twist angle (CTA), angle between clinical transepicondylar axis (CTEA) and STEA (CSA) were measured. These values were divided into different groups according to gender and side, the values of CTA, PCA, A-PA, angle PT (varus of tibia plateau), constant 3, ATA, APA and constant 90° were compared by statistically. A-PA and PCA, and CTA were analysed statistically with the liner regression, the relationship among CTEA, STEA ,PCL, APL and PLP were performed to assess by liner regression.
RESULTSATA was (89.79 ± 1.22)°, APA was (84.84 ± 1.83)°, A-PA was (5.16 ± 1.83)°, PCA was (4.80 ± 1.23)°, CTA was (8.23 ± 1.40), CSA was (3.45 ± 0.68)°. All the parameters had no differences on sex and side,but CSA had difference on male and female. There was no difference among angle PT, PCA, A-PA. There was significant difference in CAT, constant 30 and angle PT, PCA,A-PA. There was no difference between ATA and constant 90°, but there was difference between APA and constant 90°. There was relativity between PCA and CTA, and also PCA and A-PA, CTA and A-PA. There was significant relativity between STEA and CTEA, between STEA and APL, between STEA and PCL, and also between APLP, APL and PCL, but there was no significant relativity between PCL and CTEA.
CONCLUSIONTKA for Chinese, the section of femoral posterior condyle should be external 5° to obtain the optimum rotational orientation. The property is different entirely between STEA and CTEA, the rotational alignment is not performed according to parallel to the CTEA in distal femur. Among STEA, APL, PCL, the STEA is the most reliable mark ofrotational alignment of the distal femur, and the PCL is the less reliable mark.
Adult ; Arthroplasty, Replacement, Knee ; Female ; Femur ; anatomy & histology ; diagnostic imaging ; Humans ; Male ; Middle Aged ; Rotation ; Tomography, X-Ray Computed
10.Effects of mitomycin C on membrane permeability and ultrastructure changes in human pterygium fibroblasts
Xiao-xi, YANG ; Xiao-fang, CAI ; Jian, CHEN ; Bin-bin, LI ; Yan-hua, HE ; Jing-hui, ZHANG
Chinese Journal of Experimental Ophthalmology 2012;30(6):490-494
Background Mitomycin C (MMC) has an inhibitory effect on the growth and proliferation of human pterygium fibroblasts,however,there is little literature about its influence on plasma membrane. Objective This study was to investigate the influence of MMC on the physical and chemical features and ultrastructures of plasma membrane. Methods Human pterygium tissues were obtained during the surgery.Human pterygium fibroblasts were primarily cultured and passaged using explant cultured method and identified by Vimentin staining.The third generation of cells were incubated to 96 well plate at a density of 5 × 103 cells/well,and 0,50,100,200,300 and 400 mg/L MMC was added in the culture well respectively to act for 12 hours.Cell viability was assayed using cell counting kit-8 ( CCK-8 ),and cellular apoptosis was detected using annexin V-FICT/PI.The changes of cell membrane structure were examined under an atomic force microscope.Malondialdehyde( MDA ) content in cell supernatant and level of lactate dehydrogenase ( LDH ) in extracellular fluid were detected to assess the lipid peroxidation level and permeability of cell membrane.Intracellular Ca2+ changes and membrane surface topography were assessed by Fluo-3/AM mark and flow cytometry( FCM ).This study was approved by Ethic Commission of Affiliated First Hospital of Jinan University.Informed consent was obtained from each patient. Results A lot of cells grew with the shape of spindle 1-2 weeks after culture.Positive response was seen in cultured cells for Vimentin.Growth and proliferation of the cells reduced 12 hours after action of MMC with the increase of MMC concentrations.The apoptosis rate of human pterygium fibroblasts was 4.2%,4.2%,5.4%,19.3% and 25.8% in 0,50,100,200 and 300 mg/L MMC groups respectively.Different degrees of abnormalities of cells membrane were found in various concentrations of MMC groups.The elevated contents of LDH and MDA in extracellular fluid were detected in various concentrations of MMC groups compared with the control group,and the LDH and MDA levels were gradually ascended as the increase of MMC concentrations,with a significant difference between any two groups(P<0.05).The disturbance of intracellular Ca2+ homeostasis was also been seen after MMC acted. Conclusions MMC leads to the changes of physical and chemical features in human pterygium fibroblasts at a dose-dependent manner.Cell membrane may be the acting target of MMC.