Background and purpose:Breast cancer is one of the most common carcinoma among female patients with high mortalities. Drug-resistance is the major reason that leads to chemotherapy failure in clinical practice. MCF-7/ADR is a multi-drug resistant cell line that was established on the basis of breast cancer cell line MCF-7. This research aimed to investigate the anti-apoptotic effect of c-fos in resistant breast cancer cell MCF-7/ADR, and to compare with its sensitive counterpart MCF-7.Methods:Doxorubicin with various concentrations was used to treat MCF-7 as well as its MDR- counterpart MCF-7/ADR. The growth inhibitory rate of MCF-7 and MCF-7/ADR wasdetermined by MTT assay. Additionally, RT-PCR was used to test the expression of P-gp and c-fos mRNA in MCF-7 and MCF-7/ADR; The expression of c-fos mRNA was detected by RT-PCR after 3 μmol doxorubicin treatment;We further established cell lines that stably interfered with c-fos, named MCF-7/ADR/si-fos-8B, MCF-7/ADR/si-fos-3D. Flow cytometry and MTT assay were used to investigate the apoptosis rate and inhibitory rate in these above cells under the treatment of 5-FU, CDDP or γ-radiation. At last, RT-PCR and Western blot analysis were used to detect the expression of bax, bcl-2, puma, p53.Results:The expression of c-fos and P-gp (MDR-1) was up-regulated in MCF-7/ADR, compared with its sensitive counterpart MCF-7. Additionally, the resistant fold of MCF-7/ADR to doxorubicin was nearly 40; The expression of c-fos was gradually up-regulated after 3 μmol doxorubicin treatment; The sensitivity to drugs (5-FU and CDDP) was increased after c-fos interference while the apoptosis rate was also increased after 5-FU, CDDP and γ-radiation treatment. RT-PCR and Western blot analysis indicated that up-regulation ofbax,puma,p53 after c-fos interference while the expression of bcl-2 was down-regulated.Conclusion:c-fos may act as an anti-apoptotic protein in resistant breast cancer cell line MCF-7/ADR by regulating the expression of apoptosis related proteins, and may play a vital role in the formation of multi-drug resistance phenotype.

Aim To study the effect of the total flavonoids of turpinia arguta seen (TFS) on immune function of rats with adjuvant arthritis. Methods Adjuvant arthritis (AA) was induced on d0 by intradermal injection of Complete Freund′s Adjuvant (FCA), containing 10 mg heat-inactive BCG in 1ml paraffin oil. On d 28 after immunization, AA rats were killed by cervical decollation. Immune cells (splenocytes and peritoneal macrophages) were obtained, then cultured in vitro with TFS. Some immune indexes were detected respectively such as the splenocytes proliferation and the productions of IL-1 and IL-2 were measured by the method of cell proliferation, the production of prostaglandin E2 was determined by radioimmunoassay. Results Compared with normal groups, the response of splenocytes to ConA and LPS was lowered, IL-2 syn-thesis was decreased, and IL-1 and PGE_2 released from PM? were elevated. TFS (10~ -8 ~10~ -4 mg?L~ -1 ) not only increased splenocytes proliferation, which induced by ConA and LPS, and IL-2 production of splenocytes, but also reduced the elevated productions of IL-1 and PGE_2 released from peritoned macrophate in AA rats in vitro. Conculsion TFS could adjust abnormal immune function in AA rats.

Objective To investigate the changes of gastric emptying in awake rats after acute myocardial infarction(AMI). Methods The male Wistar rats were submitted to thoracotomy followed by or not by occlusion of the left coronary artery. After 24 h, AMI(n=12)and sham groups (n=9)were fed in gavage(1 ml)with ~(99m)-Tc-DTPA labeled test meal. Half gastric emptying time (GET1/2) and 50min residual rate were measured by single photon emission computed tomography. The rats of AMI group were sacrificed after 50 min. Plasma level of motilin was measured by radioimmunoassay in AMI and sham group respectively(n=9). Results GET1/2 was significantly higher in AMI group[(23.1?4.7)min] than in sham group[(16.0?4.0)min](P

Objective:To prepare and characterize specific and discrepant mouse hybridoma antibodies on membrane of HL60 and HL60/ADR cell lines.Methods:BALB/c mice were immunized by subtractive immunization induced Cp(Cyclophosphamide).McAbs were prepared by hybridoma technique,screened and detected by FACS and LSCM.Results:51 candidates and discrepant antibodies were found,and one of them (5F6) was purified and identified.Conclusion:Combination of SI with discrepant screening method should facilitate the preparing and identifying discrepant McAbs for identifying antibodies that can distinguish the differences in proteins expressed in HL60 and HL60/ADR,which is a significative and potential method in the research and target therapy associated drug-resistance.

Objective To observe the effect of inferior vena cava filter (IVCF) on prevention of bone cement implantation syndrome (BCIS). Methods Ten sheep were divided into 2 even groups, BCIS and LVCF intervention ones. First IVCF was implanted into the inferior vena cava through cervical vena-right atrium pathway under fluoroscopic monitoring to observe the influence of IVCF on BCIS. Then BCIS was es-tablished in the same sheep by compressing 10 mL of bone cement into a sheep medullary canal after mutilation of the left femur. Arterial blood pressure, heart rate, central venous pressure (CVP) and blood gas were monitored, while an ultrasonic device was utilized to monitor fat embolisms in the right atriums of the sheep. Oil red staining was performed to detect fat embolisms in pulmonary arteries after the sheep were executed. Results In BCIS group, dotted uneven resonances were found in the right atrium and right ventricle when the medullary canal pressure was increased to 120 mm Hg, indicating embolisms in the right chambers. The dotted resonances were increased to ponderous, snowflake-like ones as the medullary canal pressure climbed up. At the same time, blood pressure and Pa02 dropped significantly, the systolic blood pressure dropped to (80±11) mm Hg and PaO<.2> to the minimum 25 minutes after cone cement implantation. The heart rate and CVP increased continuously. The blood gas assay indicated respiratory and metabolic acidosis. The oil red staining showed bulk fat embolus in pulmonary arteries. But in IVCF group, the similar resonances were not observed throughout the surgery and the medullary canal pressure climbed to 400 mm Hg, reaching the maximum of our pressure gage range. The blood pressure, PaO2, heart rate and CVP did not change much compared to those before implantation. The blood gas and pulmonary oil red staining showed few changes either. Conclusion IVCF implantation can prevent the genesis of BCIS.

A subcellular proteomic method was applied to investigate the protein expression profiles of nasopharyngeal carcinoma (NPC) cell lines,CNE1 and CNE2,at various differentiation levels.Plasma membrane (PM) proteins were obtained by Percoll density grade centrifugation and subjected to twodimensional electrophoresis (2DE) followed by PDQuest software analysis.Nine proteins expressed with more than two folds difference were identified by MALDI-TOF-TOF,of which functions involved in cell differentiation,signal transduction,and metabolism.Half of these proteins,such as galectin-1 and annexin Ⅱ,were analyzed with real-time quantitative PCR or Westem blotting.We have tested a proteomic method to study differentiated NPCs at different levels and found several proteins that might be related to their biological characteristics.

Background and purpose:Multidrug resistance(MDR)is one of the major causes of progressive breast cancer chemotherapy failure.One of the major mechanisms of MDR is the overexpression of P-glycoprotein (P-gp).Therefore,the identification of novel agents which can inhibit the drug transporter function of P-gp or its expression is of utmost interest in cancer research.The aim of this study was to explore the antitumor and reversal effect of PHⅡ-7,natural products from traditional Chinese medicine(TCM).Methods:The cytotoxicity of PHⅡ7 alone and combined application of PHⅡ-7 and adriamycin(ADR)on breast cancer cells were determined using MTT assay.Annexin V–FITC/PI apoptosis detection kit was used to observe the apoptosis-inducing effect of PHⅡ-7 in MCF-7 and MCF-7/ADR cells.The effect of PHⅡ-7 on mdr1 mRNA was determined by reverse transcription PCR and real time PCR,flow cytometer was used to measure the intracellular ADR accumulation.Results:PHⅡ-7 alone inhibited cell growth of MCF-7 and MCF-7/ADR cells with the IC 50 (6.07?0.85),(5.51?1.22)?mol/L,respectively when combined with ADR,PHⅡ-7 enhanced the cytotoxicity of ADR toward MCF-7/ADR cells.In addition,PHⅡ-7 induced apoptosis both on MCF-7 and MCF-7/ADR cells;PHⅡ-7 reversed the drug resistance to ADR in MCF-7/ADR cells by inhibiting mdr1 mRNA transcription and increasing the intracellular ADR accumulation.Conclusion:PHⅡ-7 displayed significant anti-proliferative and apoptosis-inducing effect on sensitive and multidrug resistant breast cells in vitro.PHⅡ-7 reversed effectively MDR by blocking the drugs to be pumped out by inhibiting P-gp expression and function pathway.

OBJECTIVETo study the effect of butenolide (BUT) on cultured chondrocytes differentiation and the possible protective effects of selenium (Se).

METHODSEx-vivo cultured chondrocytes were divided into six groups: (1) Control group (without BUT and Se); (2) Se 0.1 microg/ml control group; (3) BUT 0.1 microg/ml group; (4) BUT 1.0 microg/ml group; (5) BUT 5.0 microg/ml group; and (6) BUT 1.0 microg/ml + Se 0.1 microg/ml group. The expression of collagen II (Col II), collagen X (ColX), basic fibroblast growth factor (bFGF), and parathyroid hormone-related peptide (PTHrP) in (or around) chondrocytes in all groups were analyzed by immunohistochemistry.

RESULTSThe expressions of Col II in 1.0 microg/ml BUT group and 5.0 microg/ml BUT group were significantly lower than those in the control group (P < 0.05). The expression of Col II in 1.0 microg/ml BUT + Se group were significantly higher than those in the 1.0 microg/ml BUT group and 5.0 microg/ml BUT group (P < 0.05). The expressions of bFGF and PTHrP of BUT groups were significantly higher than those in the Se and control groups (P < 0.05). No expression of ColX was observed in all groups.

CONCLUSIONBUT can affect the collagen II synthesis of the chondrocytes. Selenium supplementation may play a protective role.

4-Butyrolactone ; analogs & derivatives ; pharmacology ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Humans ; Protective Agents ; pharmacology ; Selenium ; pharmacology ; T-2 Toxin ; toxicity

OBJECTIVETo examine the significance of individual risk on diabetes to subjects who underwent diabetes screening.

METHODS2003 asymptomatic diabetes subjects with high-risk factors of diabetes as family history, obesity, hypertension, and/or dyslipidemia, fetal giant history were screened. 5362 subjects having no risk factors but from the same community were allocated as controls.

RESULTSThere were 131 (6.54%) diabetes identified in the screening group and 1547 (77.23%) subjects having 1 risk factor, 387 (19.27%) having 2 risk factors, 70 (3.49%) having 3 or more risk factors. There were 96 (1.79%) diabetes identified in the control group. Compared with control group, the OR (95% CI) value was 2.68 (2.20-3.25) after adjusted on age among the high risk group. The OR value of those having 1 risk factor was 2.89, but these having 3 or more risk factors increased to 4.68.

CONCLUSIONThe relation between the risk of high-risk group with diabetes and the number of risk factors of diabetes presented positive correlation. Early and regular screening for diabetes was essential in these individuals with high-risk factors.

Adult ; Aged ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; epidemiology ; etiology ; genetics ; Family Health ; Female ; Glucose Tolerance Test ; Humans ; Hyperlipidemias ; epidemiology ; Hypertension ; epidemiology ; Male ; Mass Screening ; Middle Aged ; Obesity ; epidemiology ; Odds Ratio ; Prevalence ; Risk Factors ; Smoking

OBJECTIVETo study the effect of human spastic paraplegia 21 protein (SPG21) on the replication of hepatitis B virus(HBV) and its regulatory mechanism.

METHODSHBV infectious clone pHBV1.3 and its promoter pHBV-Luc were transfected respectively into HepG2 cells with SPG21 of different concentrations, HBsAg and HBeAg in the supernatants were measured by enzyme linked immunosorbent assay (ELISA), expression of HBV core mRNA and protein were detected by RT-PCR and western blot, covalently closed circular DNA(ccc DNA) levels were measured by real-time PCR, and HBV promoter activity was measured by luminometer fluorescence detector.

RESULTSExpression of HBsAg, HBeAg, HBV core protein and cccDNA were upregulated by SPG21 as well as HBV promoter activity in a dose-dependent approach. The activity of HBV promoter increased to 1.63, 3.09 and 4.66 times in HepG2 cells treated with 50mug/ml, 100mug/ml and 200mug/ml SPG21 respectively during 48 hour-treated ( P less than 0.05), as compared to the control group.

CONCLUSIONSSPG21 can enhance the replication of HBV in HepG2 cells.

Adaptor Proteins, Signal Transducing ; metabolism ; Hep G2 Cells ; Hepatitis B virus ; metabolism ; physiology ; Humans ; Transfection ; Virus Replication