Objective To explore the Cep70 by adjusting the stability of acetylated alpha tubulin,participate in breast cancer drug resistance mechanisms.Methods (1) In order to induce taxol drug resistance cell line Michigan cancer foundation-7 (MCF-7)/pac,high-dose shock treatments taxol MCF-7 was used for 6 months,until the cells can grow in 3.5 μmol/L of paclitaxel.(2) The 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method was used to detect inhibition rate by taxol to MCF-7 and MCF-7/pac cell.(3) Immunofluorescence and Western blot were used to test acetylated alpha-tubulin and Cep70 expression levels in MCF-7 and MCF-7/pac cells.(4) Chemical intervention was used to acetylate apha-tubulin expression,Western blot and polymerase chain reaction (PCR) were used to detect the change of acetylated alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac groups.Flow cytometry and Western blot were used to detect the change of cell cycle.Results (1) IC50 of MCF-7 and MCF-7/pac was 22.47 μ mol/L and 31.38 μmol/L,respectively.(2) Immunofluorescence and Western blot results showed that the expression of acetylation of alpha-tubulin in resistant MCF-7 cell/pac was obviously decreased.(3) Real time polymerase chain reaction (RT-PCR) and Western blot showed Cep70 expression was consistent of acetylation of alpha-tubulin.(4) After incubation with paclitaxel for 24 hours,the expressions of acetylation of alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac were increased,but the extent of MCF-7 cell was much higher.Instead,incubation with nocodazole after 24 hours,the acetylation of alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac cells were obviously lowered.(5) After paclitaxel intervention,compared to the same group MCF-7 cells,the G2 phase ratio in MCF-7/pac cells was lower.In addition,given nocodazole after the intervention,compared to the same group MCF-7 cells,the ratio of G2 phase in MCF-7 cell/pac was significantly decreased.Conclusions Cep70 decreased the expression of the acetylated alpha-tubulin,reduced the stability of microtubules,which could be an important mechanism of taxol drug resistance.

Objective To reveal acetylated a-tubulin acts as pressure sensors,sensing the changes in extracellular matrix to impact on the invasion and metastasis of breast cancer.Methods The acetylated alpha microtubule protein expressions were detected,and its relationship with invasion and metastasis in breast cancer clinical specimens and different molecular subtypes of breast cancer cell lines were analyzed.The expression of acetylated a-tubulin was interfered through chemical methods,and cell morphology and the change of the invasion and metastasis ability were detected under the condition of different matrix hardness.Results The expression of acetylated a-tubulin was highest in basal-like breast cancer tissue and cell lines,and was lowest expressed in the luminal B breast cancer tissue and the breast cancer cell lines.The expression of acetylated a-microtubule was positively correlated with the occurrence of breast cancer.Under the condition of soft substrate cultivation,cell polarization was declined,becoming the circular or oval shape.The acetylated a-tubulin caused reduction in cell polarity,accompanied with less invasion and metastasis ability.Conclusions The acetylated a-tubulin acts as pressure sensors,sensing the sclerosis of extracellular matrix in the process of tumorigenesis,promoting invasion and metastasis of breast cancer.

[ ABSTRACT] AIM:To study the influence of insulin resistance on fatty liver in the mice fed with high-fat diet (HFD).METHODS:Male 8-week-old C57BL/6J mice were randomly divided into HFD group (with 60% calories by high saturated fatty acid) and control group (with chow diet).The mice in both groups were fed for 12 weeks.The body weight, liver weight, serum triglyceride (TG) and total cholesterol (TC), and blood glucose and insulin levels were meas-ured.Hyperinsulinemic euglycemic clamp experiment was applied to reflect insulin sensitivity .The lipid deposition in the liver was analyzed by HE staining , Sudan IV staining and measurement of liver fat content .The phosphorylation levels of IRS1 and Akt, and the protein levels of SREBP-1 and FAS were determined by Western blot to reflect the activities of insu-lin signaling and lipid synthesis .RESULTS:Compared with control group , the body weight and liver weight were signifi-cantly increased in HFD group .TG and TC contents in serum and liver tissues were remarkably increased in HFD group . High-fat diet induced insulin resistance , as evidenced by increased serum insulin levels , reduced glucose infusion rate and decreases in IRS1 and Akt phosphorylation levels .In livers of HFD group, HE staining showed that the cytoplasm of hepa-tocytes was filled with vacuoles .Sudan IV staining also displayed that many different sizes of red lipid drops existed in the hepatocytes , and the protein levels of SREBP-1 and FAS were significantly increased .In primary normal hepatocytes with exogenous oleic acid intervention for 48 h, the phosphorylation levels of IRS 1 and Akt were reduced , and the protein ex-pression of SREBP-1 and FAS was significantly increased in a dose-dependent manner .CONCLUSION: Feeding with HFD leads to insulin resistance , resulting in activation of lipid synthesis and accumulation of lipid deposition in the liver , thus inducing fatty liver .

Objective To evaluate the feasibility and the safety of the direct coronary stent implantation Methods Retrospectively reviewed the clinical and angiographic data of 139 patients received direct coronary stent implantation, follow up was performed in 95 of these patients Results The direct coronary stenting procedure was successful in 133 (95 7%) of 139 cases In residual 6 (4 3%) failure cases, the stent could not cross the lesion and was successful retrieved in 2 cases, balloon predilation was added In 4 cases the stent did not completely cover the lesion or dissection appeared after direct stenting, the second stent was deployed Angiographic success was achieved in all the 139 cases without major adverse coronary events during in hospital In 7 (7 4%) of 95 cases the target lesion needed repeat PTCA because of significant restenosis during 1～23 months (median 4 7 months) of clinical follow up Conclusion Direct coronary stent implantation is feasible and safe in selective patients It can save the cost of one balloon catheter

Objective To explore the characters of the aged patients with coronary disease undergoing percutaneous transluminal coronary angioplasty in clinical presentation, procedural success, and in-hospital outcomes. Methods Data were collected as a part of a prospective registry of all percutaneous coronary interventions performed by authors between January 2001 and October 2002. Comparisons between 2 age groups (≥70 years and

Objective To investigate the mechanism of glycoprotein serglycin (SRGN) promoting metastasis of breast cancer cells and the possible mechanism of SRGN expression.Methods Real time quantitative polymerase chain reaction (PCR) and bioinformation retrieval were used to detect the expression of SRGN in lymph node metastasis and non-metastasis breast cancer.MDA-MB-231 shRNA and MCF-7-SRGN of breast cancer stable cell line were established by lentivirus shRNA interferencc and overexpression.Transwell assay was used to test the effect of SRGN on invasion and metastasis of breast cancer cell line in vitro.Western blot assay was used to detect the changes of epithelial-mesenchymal (EMT) related markers.The possible regulatory mechanism of SRGN expression was detected by Western blot assay.Results SRGN expression was significantly increased in lymph node metastasis of breast cancer in clinical specimens.SRGN interference inhibited the invasion and metastasis of tumor cells.SRGN promoted breast cancer cells EMT.Transforming growth factor β1 (TGFβ1) promoted the expression of beta SRGN transcription.Conclusions SRGN can induce the change of EMT in breast cancer cells and promote the invasion and metastasis of breast cancer cells.

Objective To investigate the effects of sIL-2R in plasma on regulatory T cells (Treg) in children with Kawasaki disease ( KD ) .Methods Thirty-three children with KD and fourteen age-matched healthy children were enrolled in this study .The proportions of CD4+CD25+Foxp3+Treg cells in pe-ripheral blood and mean fluorescence intensity (MFI) of phosphorylated-STAT5 (pSTAT5) protein in CD4+CD25+T cells were analyzed by flow cytometry .The concentrations of sIL-2R, IL-2, IL-7 and IL-15 in plas-ma were measured by cytometric bead array ( CBA ) .Real-time PCR was performed to detect the gene ex-pressions of Foxp3, GITR, CTLA4, IL-2Rα, IL-2Rβand IL-2Rγat mRNA level as well as the expressions of IL-17A and ROR-γt at mRNA level in CD4+CD25-T cells.Results (1) Compared with the control group, the proportions of CD4+CD25+Foxp3+Treg cells in peripheral blood from patients with KD and the expressions of associated factors including Foxp 3, GITR and CTLA-4 at mRNA level were significantly down-regulated (P<0.05).However,the expressions of IL-17A and ROR-γt at mRNA level in Th17 cells were markedly up-regulated (P<0.05), which could be recovered to some extent after treatment with IVIG (P<0.05).(2)The expressions of pSTAT5 protein in CD4+CD25+T cells from patients with acute KD were sig-nificantly decreased (P<00.5 ), but increased with IVIG intervention (P<0.05).(3)The concentrations of sIL-2R in plasma were elevated during acute KD (P<0.05), but decreased after treatment with IVIG (P<0.05).Moreover, KD patients with coronary artery lesion ( KD-CAL+) presented a high level of sIL-2R than those without coronary artery lesion (KD-CAL-) (P<0.05), but there was no significant difference in the concentrations of IL-2, IL-7 and IL-15 in plasma between two groups (P>0.05).(4)The expressions of IL-2Rαand IL-2Rβat mRNA level in CD4+CD25+T cells from patients with acute KD were lower than those of the healthy subjects (P<0.05), but up-regulated to some extent with IVIG treatment (P<0.05).There was no significant change in the expression of IL-2Rγat mRNA level (P>0.05).The concentrations of sIL-2R in plasma were negatively correlated with the expressions of IL-2Rβand Foxp3 at mRNA level and pSTAT5 at protein level (P<0.05).The expression of pSTAT5 protein had positive correlation with the ex-pression of Foxp3 at mRNA level (P<0.05).Conclusion Aberrant IL-2/STAT5 signaling pathway media-ted by significantly increased concentration of sIL-2R in plasma might be one of the factors leading to down-regulation of Treg cells in patients with acute KD .

Objective:To evaluate the toxic effect of 3 different gingival retraction cords.Methods:DMEMextraction of DL-adren-aline HCl,aluminium sulphate and non-drug retraction cords with the extraction time of 5,10,15 and 30 min were respectively pre-pared and were used to culture human gingival fibroblasts(HGFs)in vitro respectively.Cell proliferation was tested by MTT assay. Cell apoptosis was examined by Annexin/PI method.Results:The 3 gingival retraction cord extractions inhibited the roliferation,pro-moted the apoptosis of HGFs(P <0.05),the effects were related to the extraction time.Conclusion:The 3 retraction cords have time-dependant cytotoxity.

The therapeutic effects of diltiazem in the dosage of 135~240 mg/d on 26 cas-s with coronary myocardial ischemia were observed.It was found that diltiazem could markedly slow down tho heart rate and decrease the blood pressure of the patient when he was at rest and 2,4 and 6 minutes after he underwent exercise test.It could also increase exercise tolerance,delay an exercise-induced anginal attack,improve the ischemic depression of ST segment,and PEP/LVET.Eventually it could pievent the patient from anginal attacks.The therapeutic effects of diltiazem would be more apparent in those cases of myocardial ischemia accompanied with hypertension.

Acute Disease ; Adult ; Carbon Tetrachloride Poisoning ; diagnosis ; drug therapy ; Humans ; Male