Objective: To explore the effect of bortezomib on migration and invasion of cervical carcinoma HeLa cell and specific molecular mechanism. Methods:The effect of bortezomib on the viability of HeLa cell was measured by MTT assay. The effect of bortezomib on cell migration and invasion was measured by Transwell assay and invasion experiment respectively. The activation of Akt/mTOR signaling pathway and expression level of MMP2, MMP9 were assayed by western blot. Results:MTT assay indicated bortezomib (2.5μM, 5μM, 10μM) could inhibit HeLa cell viability, and the inhibitory rate was highest at 48 h. Transwell assay and invasion experiment results showed that bortezomib inhibited HeLa cell migration and invasion. Western blotting assays presented bortezomib could suppress the phosphorylation of Akt and mTOR, and down-regulate the expression of MMP2 and MMP9. Conclusions:These results suggested bortezomib could inhibit migration and invasion in cervical carcinoma HeLa cell, which might be related to Akt/mTOR signal pathway.

Objective By investigating precision,linearity and accuracy of 9 commonly urine albumin assay systems (8 of immuno-turbidimetric assays and 1 of immuno-nephlometric assay),and comparing the concordance of measurement results,to elucidate the quality of the existing analytical systems.Methods Referring to Clinical and Laboratory Standards Institute (CLSI) EP15-A2,two mixed urines with U-Alb levels of 20 mg/L and 200 mg/L were made to validate precision; Referring to CLSI EP14-A2,fourty fresh urines were selected to evaluate matrix effect of saline diluted European Reference Materials (ERM) DA 470 and saline diluted urine,also to reflect the variation of measurement results among systems; Referring to EP6-A,saline diluted urines (10 levels) were made to validate linearity; Taking the theoretical concentration of precisely saline-diluted ERM-DA 470 as the target value,accuracy of each assay system was evaluated.Maximal allowable coefficient variation (CV) of ≤ 15％ was taken as the acceptable precision for each assay system,as rccommcnded by International Federation of Clinical Chemistry (IFCC)-and National Kidney Disease Education Program (NKDEP) ; maximum allowable bias of ≤25％ was taken as criteria for accuracy evaluation as used in Proficiency Test (PT) sponsored by College of American Pathologists (CAP).Results At level of micro-albuminuria(20-200 mg/L),all 9 systems total CVs were ≤ 15％ ; No matrix effect or interference were found in saline diluted ERM DA 470 and saline diluted urine.For A,B,E,F,G and I systems,validated linear regions were close to those stated in kit instruction;For C,D and H systems,the lower limits of validated linear region (18.7,3.6 and 12.0 mg/L,respectively) were higher than those stated in kits instruction (0,0.9 and 5.0 mg/L,respectively) ;For B and C systems,the lower limits of validated linear region were close to the upper limits of reference interval stated in kit instruction.When urine albumin was ≤ 12.6 mg/L,A,E,F,G and I systems showed good accuracy,absolute biases at all dilution were below 3 mg/L,D system showed higher positive bias (5.0-14.4 mg/L),B,C and H systems' biases were not evaluated because of high in-batch CV (the CV of B system≥ 18.1％,of C system ≥ 14.5％,of H system ≥ 39.1％); when U-Alb ranged in 25.2-201.0 mg/L,all 8 systems' relative biases were ≤25％,except D systems,which showed an un-acceptable positive bias (15.9％-44.3％).Good concordance among systems' results was present at level of microalbuminuria(20-200 mg/L),with CV among systems ＜ 15％ ;when urine albumin was ＜ 20 mg/L,CV among systems increased as allumin concentration decreased.The main contribution of variation came from B,C and H systems,which lower limits of linearity were relatively high.Conclusions At level of microalbuminuria(20-200 mg/L),except D system,the other 8 systems show good precision and accuracy;at low level of urine albumin(＜20 mg/L,especially ＜ 10 mg/L),precision and accuracy of some systems(B,C and H system) needs to be improved.

Urinary excretion of albumin indicates kidney damage and is recognized as a risk factor for progression of kidney disease and cardiovascular disease,resulting a widespread clinical utilization of urine albumin measurement.Considerable inter-method difference has been reported for urine albumin,there are no available reference materials and no reference measurement procedures for urine albumin.This review discusses the clinical utilization of urine albumin in recent years,the principles of existing measurement systems and difference of results among systems,and the current status of work about reference materials and reference measurement procedures.

Objective To explore the clinical characteristics, diagnosis, treatment and prognosis of synchronous occurrence of gastrointestinal stromal tumor( CIST) and digestive tract cancer in 6 cases. Methods The clinical and pathologic data of 6 patients of synchronous occurrence of gastrointestinal stromal tumor and digestive tract cancer from January 2005 to September 2010 in Wuhan Union Hospital were analyzed retrospectively. Results In 4 cases both tumors were located in the same organ, in the other 2 cases tumors were located in the adjacent sites of digestive tract. Only 1 case was found both tumors by preoperative examination , and the others were found digestive tract cancer. Gastrointestinal stromal tumors were all in the very low risk group, the diameter of CISTs was less than 1 cm and the mitotic score was less than 2/50 HPF in all the cases. Digestive tract cancer showed no specical demonstrations both macroscopically and microscopically. Conclusions Clinical features of synchronous occurrence of CIST and digestive tract cancer are atypical. We should pay much attention to this entity in clinical practice. In this series, most CIST are in early stage and of low risk of malignance. The prognosis of patients with synchronous occurrence of GIST and digestive tract cancer mainly depends on digestive tract cancer.

Objective To compare the influence of aging on apoptosis and the expression of apoptosis-related protein in adult and aged male Wistar rat hearts after acute coronary artery occlusion. Methods The acute myocardial infarction rat model was established by left anterior descending(LAD)occlusion.A total of 115 adult and aged rats,aged 6-24 months,were included in the study.The rats were divided into 4 groups:aged model group,aged control group,adult model group and adult control group.Animals were killed 1,3,5 hours,1 and 7 days after coronary occlusion.Hemodynamic parameters[heart rate(HR),left ventricular systolic pressure(LUSP),left ventricular end-diastolic pressure(LVEDP),±dp/dtmax)]were obtained from each group at every time points.The apoptosis and necrosis of myocardium were detected with TUNEL way and TTC stain.The expressions of Bcl-2 and Bax were analyzed with immumohistochemical stain. Results DNA fragmentation occured 1 hour after coronary occlusion and apparently peaked earlier in the aged than in the adult rat hearts.At 3 hour,the apoptotic index of aged model group was obviousily higher than that in adult model group[(51.90±23.15)%us.(18.67±17.15)%,P<0.01].The basal levels of Bcl-2 and Bax were higher in the aged than in the adult rat hearts.The expression of Bcl-2 in aged model group and adult model group were 2.7±0.9 and 1.8±0.8,P<0.05.The expression of Bax in aged model group and adult model group were 6.2±2.9 and 4.2±1.5,P<0.05. Conclusions The ability of aged rats to resist ischemia is poor,aging may alter the expressions of Bcl-2 and/or Bax,increase cardiomyocyte apoptosis,thereby,enhance the myocardial dysfunction during acute myocardial infarction.

BACKGROUND:Osteoporosis and its complications severely threaten the elder’s health. Simvastatin, widely accepted as a lipid-lowering drug, is reported to potentialy promote bone formation, but it is in debate when oraly administered, and there is no evidence to support whether this is due to the region difference.
OBJECTIVE:To investigate the effect of oraly administered simvastatin on bone mass and biomechanical properties of the femur and vertebrae in osteopenia rats induced by ovariectomy (OVX).
METHODS: Twenty-four 6-month-old female Sprague-Dawley rats were subjected to OVX+oraly administered saline vehicle (OVX group,n=8), OVX+oraly administered simvastatin (5 mg/kg/d; intervention group,n=8) or sham surgery (sham group,n=8). After 8 weeks of treatment, al rats were sacrificed and the level of procolagen type I N-terminal propeptide in blood serum was assessed by ELISA. Bone mineral density was determined in the L5 vertebra and left femur using dual-energy X-rays. Furthermore, the biomechanical properties of the L4 vertebra and right femur, including maximum load and elastic modulus, were detected by compression testing and three-point bending test, respectively.
RESULTS AND CONCLUSION: The serum level of procolagen type I N-terminal propeptide in the sham group was significantly lower than that in the other two groups. OVX rats showed significantly lower bone mineral density in both the L5 vertebra and left femur than sham rats (P < 0.05). Rats in the intervention group showed higher bone mineral density than those in the OVX group, with statisticaly significant difference in the L5 vertebra (P < 0.05), but insignificant difference in the femur. Maximum load and elastic modulus of the L4 vertebra in the OVX group were significantly lower than those in the sham and intervention group. Markedly lower elastic modulus of the femur was found in the OVX group than the sham and intervention groups. These findings demonstrate that simvastatin treatment can partialy prevent bone loss in OVX rats with more notable effect on the vertebrae than the femur, and for this model, the vertebra is superior to the femur used in biomechanical test.

Objective To explore the influencing factors of gastrointestinal bleeding(GB) in secondary prevention patients with cerebral ischemic stroke(CIS).Methods A total of 616 patients were divided into bleeding group and control group according to the status (yes,no) of suffering GB during the 2years follow-up.Single factor analysis and Logistic regression analysis was used to explore the influencing factors of GB in CIS patients.Results The proportion of age≥65,a history of GB,gastric disease,renal insufficiency,sudden onset,NIHSS ≥12 and CIS ≥2 in the bleeding group was significantly higher than that in the control group (P ＜ 0.05) ; The proportion of combined with statins,proton pump inhibitors and gastric mucosal protective agent in the bleeding group was significantly lower than that in the control group (P ＜0.05) ; The Logistic regression analysis showed that age≥65,a history of GB,gastric disease,renal insufficiency,sudden onset,NIHSS≥12 and the times of CIS≥2 were risk factors of GB; however,combined with statins,proton pump inhibitors were protective factors.Conclusions Aging,a history of GB,gastric disease,sudden onset,higher NIHSS score and the times of CIS ≥ 2 were the risk factors of GB,combined with statins and proton pump inhibitors could reduce the risk of GB.

The effects of estradiol and tamoxifen on the proliferation of estrogen receptor positive cells and the relationship between the tamoxifen tolerance and cell origin were investigated. The tissues of human endometrium and breast cancer were randomly selected following dissection for primary cell culture. After the breast cancer cells and endometrial cells were treated with 1 x 10(-8) mol/L estradiol and/or 1 x 10(-6) tamoxifen, 3H-labelled thymine nucleotide was used to trace the kinetics of cell proliferation. There was no significant difference in the inhibition on the human endometrial cells between tamoxifen-treated group (6.3%) and control group (6.4%), but tamoxifen could significantly inhibit the proliferation of the human breast cancer cells (45.84%) as compared with control group (52.72%). Moreover, tamoxifen could significantly stimulate the proliferation of tamoxifen resistant breast cancer cells (9.64%) as compared with control group (6.32%). Estradiol could significantly stimulate the proliferation of all the three kinds of cells as compare with control group. The combined use of estradiol and tamoxifen could inhibit the proliferation of the endometrial cells and breast cancer cells as compared with estradiol used alone, but on the tamoxifen resistant breast cancer cells, they could more significantly stimulate the proliferation than E2. It was concluded that E2 could stimulate the proliferation of these three kinds of cells. However, the inhibitive effects of tamoxifen on the proliferation of these cells were dependent on the estradiol.
Antineoplastic Agents, Hormonal/*pharmacology ; Breast Neoplasms/*pathology ; Cell Division/drug effects ; Cells, Cultured ; Drug Interactions ; Endometriosis/pathology ; Endometrium/*pathology ; Estradiol/*pharmacology ; Tamoxifen/*pharmacology ; Tumor Cells, Cultured

Objective To explore the effects of continuous renal replacement thempy ( CRRT) on efficacy in severe acute pancreatitis ( SAP) patients based on different time .Methods A total of 38 SAP patients were divided into two groups randomly:early group(CRRT within 48h) for 21 patients and late group(CRRT after 48h)for 17 pa-tients.the APACHE II score,the length of stay ICU and the mortality of 28d were compared between the two groups . Results The APACHE Ⅱscore was significantly higher in late group than that in early group in day 1 and 2 ( t=2.231,2.304,both P<0.05);there is no significant differences between late group than that early group in APACHEⅡscore before treatment(P>0.05);the hospitalizing days and mortality were significantly higher in late group than that in early group(t=2.156,χ2 =4.293,both P<0.05);there was no significant differences between late group and early group in gender,age,APACHEⅡscore(all P>0.05).Conclusion Early CRRT can decrease both the hospi-talizing days and mortality in SAP patients .CRRT may provide more clinical benefits in the early phase (within 48h) of SAP.

Objective To investigate the effect of dexmedetomidine (DEX) on proliferation and differentiation of rat embryonic neural stem cells in vitro. Methods Embryonic neuraI stem cells of fetal SD rats were separated from rats with gestational age of 14-16 days , and underwent primary culture for nestin expression. Cells were divided into control group, low-dose DEX group, and high-dose DEX group, which were cultured with 0,1 and 10 ng/mL DEX respectively. Cell viability was detected by MTT assay;the proliferation rate was estimated by BrdU incorporation; the phosphorylation of ERK1/2 in the proliferation phases was detected by western blot; the differentiation of neuronal cells,astrocytes,oligodendrocyte and neural stem cells were assessed by immunocytochemistry of cell specific markers. Results More than 95% of the embryonic neural stem cells in primary culture were Nestin-positive. Cell viability , BrdU-positive cells and phosphorylation of ERK1/2 in low-dose DEX group increased significantly (P<0.05), and decreased significantly in high-dose DEX group compared with control group and low-dose DEX group (P < 0.05). NeuN-, GFAP- and MBP-positive cells increased (P < 0.05), while Nestin-positive cell decreased (P<0.05) in both low and high dose DEX group compared with control group. However, no significant difference was found in low and high dose DEX group (P > 0.05). Conclusions The proliferation of neuraI stem cells can be promoted by low-dose DEX and depressed by high-dose DEX , as well as regulated by DEX of the phosphorylation of ERK1/2. DEX induces neural stem cells non-selective differentiation.