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 establish an HPLC method to simultaneously determine the contents of shikonin andβ,β'-dimethylacrylshikonin inZizhu Ointment.Methods The contents of shikonin andβ,β'-dimethylacrylshikonin inZizhu Ointment were determined by HPLC. The chromatographic conditions were Welch Materials XB-C18 Column (4.6 mm × 250 mm, 18.5μm), with mobile phase of acetonitrile-water-formic acid (75:25:0.05), detection wavelength of 516 nm, temperature of 30℃, flow rate of 1.0 mL/min and the load of 20μL.Results Shikonin showed a good linearity in the range of 0.025 6-0.256μg (r2=0.999 5), and the average recovery rate was 98.12% (RSD=1.92%).β,β'-dimethylacrylshikonin showed a good linearity in the range of 0.174 4-2.616μg (r2=0.999 2), and the average recovery rate was 100.46% (RSD=1.17%).Conclusion This method is simple and accurate, which can be used for the quality control and evaluation of Zizhu Ointment.

One hundred and eighty two outpatients with functional dyspepsia (FD)were enrolled.The cognitive questionnaire and the short form Nepean Dyspepsia Index (NDI) were used for survey.Spearman analysis showed that the cognitions of symptoms affected by emotion (r =0.284,P =0.006),somatisation symptoms induced by other diseases (r =0.211,P =0.045),and fears of cancer (r =0.217,P =0.039) were positively correlated with NDI scores.Multiple linear regression analysis demonstrated that cognition of symptoms affected by emotion (β =3.709,P =0.009),somatisation symptoms induced by other diseases (β =3.259,P =0.020),need of hospitalization (β =4.533,P =0.006),and need of medication for several years (fβ =-3.207,P =0.029) were associated with NDI scores.These results suggest that the quality of life might be effected by cognitive factors,and the correction of cognitive mistakes may improve quality of life of FD patients.

BACKGROUND:Tissue inhibitor of matrix metaloproteinase 1 (TIMP-1) is the corresponding antagonist of matrix metaloproteinase 13 (MMP-13), and their balance between expression and functional activity exerts an important role in the metabolic state of the extracelular matrix. During the development of osteoarthritis, however, TIMP-1 and MMP-13 expressions and their expression ratio show unclear changes in DH guinea pigs.
OBJECTIVE:To explore the expression levels of MMP-13 and TIMP-1 in DH guinea pigs with different ages, and to analyze the relationship between the ratio of MMP-13 to TIMP-1 and the age-dependent degenerative changes in the articular cartilage.
METHODS:Twenty-four female Dunkin Hartley guinea pigs were sacrificed at age of 2, 4, 8, 12 months separately, with six animals at each time point. The knee joints were colected and gross visual appearance of the articular cartilage was observed, then were decalcified and prepared for paraffin sections. VG staining and Mankin score were used to analyze the histological changes. Immunohistochemistry was conducted to assess the expression levels of MMP-13 and TIMP-1 in the cartilage. Integrated absorbance values were used as the quantitive analysis calculated by Image pro-Plus 6.0. Linear regression analysis was done to analysis the relationship between Mankin score and the ratio of MMP-13/TIMP-1. RESULTS AND CONCLUSION:Normal appearance in the articular cartilage was observed in 2-month-old DH guinea pigs, while degenerative changes in the articular cartilage were shown in 4-month-old animals, which became severer with age. Significant difference was found in Mankin score between any two groups (P < 0.05). The ratio of MMP-13 to TIMP-1 increased with age, and the ratio was positively correlated to the Mankin score (P < 0.05). Age-related articular cartilage degeneration occurred in Dunkin Hartley guinea pigs at 4 months of age, and devoloped with age, which is related with the imbanlance of the expression ratio of MMP-13 to TIMP-1.

Objective To evaluate the effect of conversion from mycophenolat mofetil (MMF) to enteric-coated mycophenolate sodium (EC-MPS) on gastrointestine-related quality of life,as well as the safety and efficacy.Method A total of 41 renal transplant (RT) recipients were converted from MMF to EC-MPS (46.3 ± 17.1) months after the operation due to the gastrointestinal side effects of MMF,with a mean time of 46.3 months.Before the conversion and 12 weeks later,the patients were evaluated with Gastrointestinal Quality of Life Index (GIQLI) questionnaire,and the safety and efficacy were assessed.Result The average dose of MMF was 846.7 ± 291.3 mg/day before the conversion,with a mean dose of 639.5 ± 186.4 mg/day for EC-MPS.The total score of GIQLI was 103.6 ± 10.7 before the conversion,and 12 weeks after conversion 118.3 ± 15.1,with a statistical significance (P＜0.05).The safety of EC-MPS was excellent without infection,acute rejection episode,loss of allograft or death.The serum creatine was 136.9 ± 35.7 mol/L before conversion and 128.4± 40.8 mol/L after conversion (P ＞ 0.05).Conclusion For the RT patients with gastrointestinal side effects of MMF,conversion to EC-MPS could significantly alleviate gastrointestinal illness,and improve quality of life,with excellent safety and efficacy.