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OBJECTIVE:To study the effects of podophyllotoxin derivative QW-83 on human cervical cancer HeLa cell apopto-sis and its mechanism. METHODS:After treated with 0(negative control),0.01,0.1,1 and 10 μmol/L QW-83 and positive drug etoposide(VP-16)for 48 h,proliferation inhibition rate and IC50 of HeLa cell were determined by MTT assay. The morphological changes of HeLa cell were observed by Hochest 33342 staining after treated with QW-83 [0(negative control),2.5,5,10μmol/L] for 48 h;flow cytometry was used to detect apoptosis rate;semi quantitative RT-PCR was adopted to detect the expression of apop-tosis related gene P53,Bax,Casepase-3,Casepase-8,Casepase-9 and Bcl-2 mRNA. RESULTS:Compared with negative control, 1,10 μmol/L VP-16 and QW-83 had obvious proliferation inhibition effect on HeLa cells (P<0.05 or P<0.01),and IC50 were (5.11±0.43)μmol/L and(4.96±0.54)μmol/L. Hochest 33342 staining results showed QW-83 could obviously induce cells apopto-sis and nuclear pyknosis. Flow cytometry showed QW-83 could increase apoptosis rate in concentration-dependent manner,being 16.89%-62.56%. RT-PCR showed mRNA expression of P53,Bax,Caspase-3,Casepase-8 and Casepase-9,Bcl-2/Bax increased, while mRNA expression of Bcl-2 decreased after treated with QW-83(P<0.05). CONCLUSIONS:Podophyllotoxin derivative QW-83 can induce HeLa cell apoptosis,and its mechanism may be associated with regulate mRNA expression of apoptosis related gene.
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Objective To analyze the features of hidden blood loss in the elderly femoral intertrochanteric fractures treated with extramedullary dynamic hip screw (DHS), intramedullary nails (Gamma 3, PFN) and hip arthroplasty (LBFH). Methods The clinicle records of 193 elderly patients (ages ≥75 year old) with femoral intertrochanteric fractures treat by DHS,Gamma3, PFNA and LBFH in our hospital were retrospectively analyzed. The estimated blood loss were calculated by Gross equation, according to the height,weight and changes of blood routine test reoperative and postoperative,the differences of hidden blood loss among DHS group,Gamma 3 group, PFN group and LBFH group were compared. Results Total blood loss in intramedullary nail groups were significantly higher than DHS group [(766 ± 83) mL],P 0.05), The hidden blood loss were significantly higher in the intramedullary nail groups than those in the LBFH group [(453 ± 98) mL,accounting 54%] and DHS group [(429 ± 59) mL,accounting 56%] (P 0.05). Conclusion Noteworthy, the hidden blood loss is major part of perioperative total blood loss in the elderly femoral intertrochanteric fractures , intramedullary fixations may result in greater hidden blood loss than extramedullary fixations and hip arthroplasty ,which may cause postoperative anemia.
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<p><b>OBJECTIVE</b>The purposes of our study were to investigate the association between maternal urinary phthalate metabolites and the levels of inhibin B (INHB) and insulin-like factor 3 (INSL3) in the cord blood in a Chinese pregnant population.</p><p><b>METHODS</b>Maternal urine samples in the third trimester of pregnancy of 69 participants were collected and stored, and the samples of cord blood (10 ml) were collected at delivery between June 2011 and September 2012 in a comprehensive hospital of gynecology and obstetrics in Tianjin, China.Four phthalate metabolites, monomethyl phthalate (MMP), monoethyl phthalate (MEP), monobutyl phthalate (MBP), and mono-2-ethylhexyl phthalate (MEHP) were measured in the urine samples using liquid chromatography-tandem mass spectrometry. The levels of INHB, INSL3 in the cord blood were tested by ELISA. Associations of phthalate exposure with INHB and INSL3 levels were determined by spearman correlation and multiple regression model analysis.</p><p><b>RESULTS</b>The median concentrations of observed metabolites in descending order were 49.74 µg/L for MMP, 24.96 µg/L for MEHP, 19.52 µg/L for MEP and 17.73 µg/L for MBP. The median concentrations of INHB and INSL3 were 89.09 and 106.21 ng/L.Significant negative associations between INHB and MMP(β' = -0.252), MEP(β' = -0.363) or the sum value (∑PAEs) (β' = -0.346) were found by the multiple regression model analysis. For INSL3, only the sum value (β' = -0.313) was inversely significantly associated with the levels of INSL3 in the cord blood.</p><p><b>CONCLUSIONS</b>Maternal urinary phthalate metabolites were associated with INHB and INSL3 in the cord blood in a Chinese population.</p>
Subject(s)
Adult , Female , Humans , Infant, Newborn , Male , Pregnancy , Young Adult , Diethylhexyl Phthalate , Urine , Fetal Blood , Chemistry , Inhibin-beta Subunits , Blood , Insulin , Blood , Maternal Exposure , Phthalic Acids , Urine , Proteins , Testicular Hormones , BloodABSTRACT
Objective:To investigate effects of viola on macrophage TOLL-like receptor expression ,and tentatively explore the partial mechanism of viola intervention on macrophage function.Methods: Using viola water decoction lavage intervention in clean grade SD rats of conventional preparation containing serum ,In a certain concentration coculture with mouse peritoneal macrophages in 96-well plates.After 24 hours,cell phagocytosis activity was determined by neutral red uptake assay ,changes in expression levels of TLR-1,TLR-2,TLR-3,TLR-4,TLR-5 mRNAs were determined by RT-PCR.Results:Compared with the same concentration of normal serum group :(1 ) viola-containing serum group macrophage activity was significantly increased ( P0.05 ) ,and the expression level of TLR-5 was significantly increased in each group ( P<0.01 ).Conclusion:Viola-containing serum has obvious promoting effect on macrophage phag-ocytosis,the mechanism of which may be related to that this drug regulates and controls part of macrophage TOLL -like receptor expres-sion.
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<p><b>OBJECTIVE</b>To observe the effects of dibutyl phthalate (DBP) and monobutyl phthalate (MBP) on the mRNA and protein expression of insulin-like factor 3 (INSL3) in the Leydig tumor cells (MA-10) of mice and the level of testosterone secreted from MA-10 cells.</p><p><b>METHODS</b>The MA-10 cells of mice, used as a cellular model, were exposed to DBP and MBP. The content of testosterone in the supernatant medium was measured by enzyme-linked immunosorbent assay; the mRNA and protein expression levels of INSL3 in MA-10 cells were measured by quantitative PCR and Western Blot.</p><p><b>RESULTS</b>Compared with the control group, MA-10 cells showed increased synthesis of testosterone when exposed to low concentrations of DBP and MBP (10(-9) ∼ 10(-6) mol/L) and inhibited synthesis of testosterone when exposed to high concentrations of DBP and MBP (10(-3) mol/L), and the typical two-way effects became more significant as the time went one and the concentrations increased (P < 0.05). Compared with the control group, MA-10 cells showed significantly lower mRNA and protein expression levels of INSL3 when exposed to 10(-6) and 10(-4) mol/L DBP (P < 0.05); MA-10 cells showed increased protein expression of INSL3 when exposed to 10(-7) mol/L MBP, and the mRNA and protein expression levels of INSL3 decreased as the concentration of MBP increased.</p><p><b>CONCLUSION</b>DBP and MBP can inhibit the secretion of testosterone from MA-10 cells at high concentrations, but stimulate the secretion of testosterone at low concentrations. Both DBP and MBP have inhibitory effects on the mRNA and protein expression of INSL3 in MA-10 cells.</p>
Subject(s)
Animals , Male , Mice , Cell Line , Dibutyl Phthalate , Toxicity , Insulin , Metabolism , Leydig Cells , Metabolism , Phthalic Acids , Toxicity , Proteins , Metabolism , Testosterone , Bodily SecretionsABSTRACT
The aim of this study was to evaluate the effects of low concentrations of DEHP and MEHP on steroidogenesis in a murine Leydig tumor cell line (MLTC-1) in vitro. The result of flow cytometry analysis revealed that the proportion of apoptotic cells was significantly increased after the exposure to DEHP. All three genes (P450scc, P450c17, and 3βHSD) under study showed an increased expression following exposure to DEHP or MEHP, although some insignificant inhibitory effects appeared in the 10 μmol/L treatment group as compared with the controls. It was also found that compared with the controls. It was also found that DEHP or MEHP stimulated INSL3 mRNA and protein especially in the 0.001 μmol/L treatment group. Testosterone secretions were stimulated after the exposure to DEHP or MEHP. Alternations of steroidogenic enzymes and INSL3 in MLTC-1 cells might be involved in the biphasic effects of DEHP/MEHP on androgen production.
Subject(s)
Animals , Mice , Apoptosis , Cell Line, Tumor , Diethylhexyl Phthalate , Toxicity , Leydig Cell Tumor , Metabolism , Pathology , SteroidsABSTRACT
Objective: To study the effect of new photosensitizer Chlorophyl-derivative (CPD4) combined with Adriamycin on cell cycle and cell proliferation of MCF-7 breast cancer cell line and to investigate the mechanism of combination therapy. Methods: A new type of photosensitizer and traditional chemotherapy drug Adriamycin (ADM) were used to treat breast cancer cell line MCF-7. Flow cytometry was employed to detect apoptosis rate and cell cycle distribution in ADM group, group with photodynamic effect and the combined group. The influence of ADM on the mean fluorescence intensity and the changes in the mean fluorescence intensity after CPO4 (1.5μg/mL) treatment were analyzed. Results: The apoptosis rate of the combination group was higher than that in the other two groups, with statistical significance (P＜0.01). Photodynamic effect caused G_0/G_1 phase arrest in MCF-7 cells. Low concentration of ADM increased the number of G_2/M phase cells. The percentage of G_2/M phase cells was increased in the combination group. No significant difference was found in the mean fluorescence intensity between ADM pretreated MCF-7 cells for 24 hours and 48 hours and the control group (P＞0.05). Pretreatment of MCF-7 cells with ADM increased the volume of photosensitizer CPD4 into the cells. The mean fluorescence intensity at 2 hours after CPD4 incubation was the highest. Conclusion: ADM can increase the amount of CPD4 into the MCF-7 cells. Photodynamic therapy com-bined with Adriamycin has a synergistic effect on MCF-7 cells.
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Response Evaluation Criteria In Solid Tumors (RECIST) and WHO Criteria are the most in portant ones.There are some differences between them,in one word,RECIST is better than WHO Criteria when implemented in clinic.However,there are some shortcomings in RECIST.Improvement is needed to make the criteria more and more Derfect and scientific.