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.

Objectives:To observe the role of combined use of early micro feeding and intravenous nutrition in low birth weight(LBW) infants. Methods:Fifty four cases of LBW infants were randomly divided into two groups.Early micro feeding and intravenous nutrition were given in one group(EF & IN group) and another group(Control group) was given only with intravenous nutrition.The times of intravenous nutritional support requirement and hospital stay,the increase in body weight and the changes in serum bilirubin,lipids and creatinine were compared between the two groups. Results:The times of intravenous nutritional support and hospital stay were shortened and the body weight was increased in EF & IN group. The levels of serum bilirubin and creatinine and the serial concentrations of lipids on the day 7 and 14 after birth were significantly lower than those in control group. Conclusions:The combination of early,micro feeding and intravenous nutrition is superior to the only use of intravenous nutrition in shortrenning the critical course,increasing the body weight,beginning the oral intake and decreasing the possible injuries from total parenteral nutrition.

Objective To investigate the optimal b value of diffusion-weighted imaging (DWI)in predicting the grade of cerebral gliomas.Methods 38 patients with pathologically-proved brain gliomas (24 high-grade gliomas and 14 low-grade ones)were studied retrospectively.All patients received conventional MRI and DWI examination with three different b values (1 000 s/mm2 ,2 000 s/mm2 and 3 000 s /mm2 ).The signal features of the tumor on DWI with three different b values were evaluated and compared.The minimum ADC values of the tumors were calculated and compared between each other.Through drawing the ROC curve of different b values, the best diagnostic threshold was found.The sensitivity and specificity in predicting the grade of brain gliomas were assessed using Chi-square test.Results On DWI with b value of 3 000 s/mm2 ,91.6% (22/24)of high-grade tumors showed hyper-intensity, while 85.7%(12/14)of low-grade tumors presented hypo-intensity signals.Regarding hyper-intensity signals as diagnostic criterion for high-grade tumors,the sensitivity and specificity were 91.6% and 100% respectively.When the b value was similar,the minimum ADC value of high-grade glioma was significantly lower than that of low-grade glioma,and statistically significant differences in differentiating high-grade glioma from low-grade glioma existed among three b values (P <0.05).When the ADC 3 000 value<0.74×10 -3 mm2/s was regarded as a standard for the identification of high-grade and low-grade glioma,the sensitivity and specificity were 100% and 87.3%respectively.Conclusion DWI with high b value is more useful than standard moderate b value in preoperative grading the gliomas.When DWI with b value of 3 000 s/mm2 is used,the minimum ADC value will provide quantitative indicators in preoperatively precisely predicting grading glioma.

Objective To explore the mechanism of intrauterine growth restriction (IUGR) via observing the change of mitochondria in IUGR placenta. Methods Placenta samples were collected from 30 singleton pregnancies at the time of elec-tive caesarean section. Fifteen of them were appropriate for gestational age and 15 were IUGR. Mitochondrial morphology was observed by transmission electron microscopy, DNA copies were analyzed by real-time quantitative PCR and membrane potential was assayed by lfow cytometry. Results Signiifcant morphological changes of placental mitochondria were observed under transmission electron microscopy in IUGR, mitochondrial DNA copies in IUGR placenta were signiifcantly increased (P<0.01) and membrane potential decreased dramatically (P<0.01). Conclusions It is suggest that impaired mitochondrial function in IUGR may involve in IUGR pathogenesis.

Aim To investigate the effects of panax nonstaining saponins ( PNS ) on the apoptosis of renal cells induced by cisplatin through the path of mitochon-drion . Methods Male Sprague-Dawley( SD) rats were randomized divided into normal control group, cisplatin model group and the cisplatin+PNS group,with 12 rats of each group. Animals were sacrificed to determine the N-acetyl-β-D-Glucosaminidase ( NAG ) in urine, blood urea nitrogen ( BUN) and serum creatinine ( Cr) concentrations after 8d of intraperitoneal injection. HE-staining was employed to observe renal pathologies. Transmission electron microscopy ( TEM ) was em-ployed to observe the mitochondria of the rats′ injured kidney region. TUNEL staining was employed to detect the distribution of apoptotic cells. Immunnohistochem-istry was used to detect Bax and caspase-9 expression, and expression of Bcl-2 protein was detected by West-ern blot. Results Compared with the normal control group, contents of BUN, serum Cr and urinary NAG levels of rat in the cisplatin model group were increased ( P <0. 01 ) , and some mitochondria of the epithelial cells of renal tubules got injured seriously. The apopto-sis rate of kidney cell was increased ( P<0. 01 ) . The expression of Bax,caspase-9 and Bcl-2 proteins was in-creased ( P < 0. 01 ) . Compared with the cisplatin model group, contents of BUN, serum Cr and urinary NAG levels of rats in the cisplatin model group were decreased ( P <0. 01 ) , and some mitochondria of the epithelial cells of renal tubules were significantly im-proved. The apoptosis rate of kidney cell was decreased ( P<0. 01 ) . The expression of Bax and caspase-9 pro-tein was decreased (P<0. 01),but Bcl-2 protein was increased ( P < 0. 01 ) . Conclusion PNS may in-crease the expression of Bcl-2 protein and decrease the expression of Bax and caspase-9 proteins, which may play a protective role in cisplatin nephritic damage.

Objective To observe the effects of rosuvastatin on the homocysteine (Hcy)-induced expression of matrix metalloproteinase 2 (MMP 2) and cell migration in rat vascular smooth muscle cells (VSMCs), and to explore the possible mechanism of Hcy-induced atherosclerosis and the role of statins in reversing atherosclerosis. Methods In one cell culture plate, the cultured rat VSMCs were incubated with different concentrations of Hcy (0, 50, 100, 500, 1000 μmol/L and 5000 μmol/L) in vitro for 24 h, 48 h and 72 h. And in another cell culture plate, the different concentrations of rosuvastatin (10-9, 10-8, 10-7, 10-6, 10-5 mol/L and 0 mol/L) were added to the cultured rat VSMCs (while the concentration of Hcy was 1000 μmol/L). The MMP 2 expression and enzyme activity were determined by gelatin zymography and Western blotting. The effects of Hcy and rosuvastatin on cell migration and invasiveness of VSMCs were observed. Results Hcy (50-5000 μmol/L) increased the protein expression, and Hcy (50-1000 μmol/L) increased enzyme activity of MMP 2 significantly. But Hcy (5000 μmol/L) inhibited activity of MMP 2 (F=9.31, 6.44 and 5.97, all P＜0.05). Rosuvastatin (10-9-10-5 mol/L) inhibited Hcy-induced expression and enzyme activity increasing of MMP 2. The counts of cell migration of VSMCs were 18.32±2.17, 32.68±4.34, 44.75±4.08, 61.39±5.21, 79.74±5.54 and 90.78±5.83, while the concentration of Hcy was 0, 50, 100, 500, 1000 μmol/L and 5000 μmol/L respectively (F=5.31, P＜0.05). The counts of cell migration of VSMCs were 79.74±5.54, 62.53±6.41, 48.37±5.66, 31.41±4.79, 19.27±3.62 and 11.17±2.33, while the concentration of rosuvastatin was 10-9, 10-8, 10-7, 10-6 and 10-5 mol/L respectively (F=4.99, P＜0.05). Rosuvastatin could decrease the stimulation of Hcy-induced migration of VSMCs. Conclusions Hcy can influence the MMP 2 protein expression/activity in VSMCs, and rosuvastatin can inhibit augmentation of Hcy-induced MMP 2 expression/activity and migration of VSMCs. It may be one of the multiple-effects of rosuvastatin reducing atherosclerosis.

Objective To explore the expression of 11-β hydroxysteroid dehydrogenase 2 (11-β HSD2) gene in placenta of pregnancy induced hypertension (PIH) complicated by intrauterine growth retardation (IUGR) and the relationship between different expression of 11-β HSD2 in placenta and newborn's birth weight or placental weight. Methods Thirteen cases of term pregnancy mothers with PIH complicated by IUGR were served as PIH complicated by IUGR group, 22 cases of term pregnancy mothers complicated by PHI with appropriate for gestational age (AGA) infant as PIH with AGA group and 36 cases of normal controls as control group. The mRNA expression level of 11-β HSD2 gene in placenta was evaluated by RT-PCR. The level of cord serum cortisol was detected by the method of chemiluminescence. Results The 11-β HSD2 gene mRNA was expressed in placenta. The mRNA expression level of 11-β HSD2 gene in PIH complicated by IUGR group's placenta was significantly lower (0.26±0.09) than that in PIH with AGA group (0.64±0.19) and control group (0.66±0.20). The level of cord serum cortisol in PIH complicated by IUGR group was significantly higher [(71.60±20.20)μg/L] than that in PIH with AGA group [(51.00±13.80)μg/L] and control group [(49.10±14.40)μg/L]. The newborn's birth weight and placental weight in PIH complicated by IUGR group was significantly lower than those in PIH with AGA group and control group. The mRNA expression level of 11-β HSD2 gene in placenta was positively correlated with the birth weight of their newborns and placental weight. Conclusion The lower mRNA expression level of 11-β HSD2 gene in placenta may contribute to the higher cortisol level in fetal of PIH complicated by IUGR and has a negative role on the fetal development.