Objective:To explore the role and mechanisms ofω-3 polyunsaturated fatty acids (ω-3PUFAs) alone or in combination with dexamethasone (DEX) in inducing cell apoptosis and reversing drug resistance in multiple myeloma (MM). Methods:DEX-resistant MM cell line MM1R was treated with different concentrations of eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) alone or in combination with DEX for 24 or 48 h. Cel proliferation was detected by MTT assay. Cel cycle and apoptosis were measured by flow cytometry. Expression levels of apoptosis-related proteins were analyzed by Western blot. Two-tailed, unpaired Student's t-test was used to compare the two treatment groups. A value of P<0.05 was considered statistically significant. Results:MM1R proliferation was inhibited by different concentrations (10, 20, 50, and 100μM) of EPA or DHA alone or in combination with 10μM DEX in a dose-and time-dependent manner. The inhibition effect was significantly higher in combinative groups than in single EPA or DHA treatment group (P=0.014, P=0.032). The percentage of G0/G1 phase and cell apoptosis rate in MM1R treated with different concentrations of EPA or DHA alone increased in a dose-dependent manner. This percentage was also significantly higher in the combinative groups than in the single EPA or DHA treatment group (P=0.015, P=0.004). The expression levels of cleaved caspase-3 and Bax were upregulated, whereas those of pro-caspase-3 and BCL-2 were downregulated in a dose-dependent manner. Drug resistance gradually decreased in MM1R cells at different concentrations of EPA or DHA with the increase of drug concentration. The reversal fold also increased gradual y, whereas the cel s decreased in the two drug-combination groups compared with the single-drug group. Moreover, the drug-resistance reversal index increased significantly. Conclusion:ω-3PUFAs can inhibit DEX-resistant MM cell proliferation, arrest cell cycle, and induce cell apoptosis.ω-3PUFAs also exhibit a synergistic anti-resistanteffect in combination with DEX. Furthermore,ω-3PUFAs can serve as novel effective drugs for MM treatment.

Objective Recent.studies have found a strong association of insulin resistance, which might occur during ischemia reperfusion in vitro in the experimental dogs, with disturbed function of cardiomyocytes. Obvious acute insulin resistance, along with glucose dysmetabolism in the reperfused cardiomyocytes, was furher observed in the study performed with ischemia-reperfused ventric- ular myocytes of rats. We tried to investigate preliminarily the molecular mechanisms of insulin resistance in the cardiomyocytes after ischemia reperfusion. Methods An experimental model of insulin-stimulated ischemia reperfusion (SI/R) was created by isolating cardiomyocytes from adult rats. Glucose uptake of the cardiomyoctyes was evaluated with isotope-labeling technique. Glucose trans- porter 4 (GLUT4) translocation induced by insulin was investigated with Western blot analysis, and the intracellular level of free Ca2+ ([Ca2+]I) was measured quantitatively with Ca2+ indicator Fura-2. Results Insulin can stimulated glucose uptake by cardiomyo- cytes, indicating that these cells were insulin-sensitive. Cardiomyocytes were demonstrated notable acute insulin resistmce during reperfusion. Insulin-stimulated GLUT4 translocation in the cardiomyocytes 15 minutes after reperfusion was 72.2% of that in the con- trol group(P<0.05), in which the GLUT4 content in plasma membrane remained unchanged. The finding suggested that a disturbed GLUT4 translocation might happen in the cardiomyocytes during insulin-stimulated ischemia-reperfusion. Calcium overload was identi- fied in the cardiomyocytes with ischemia reperfusion. At 15 minutes of reperfusion, [Ca2+]I was significantly higher in the reperfused cardiomyocytes than that in the control cardiomyocytes[(318.66±23.06)vs(130.70±0.82) nmol/L, P<0.05], and kept at a higher level [(177.79±17.46) nmol/L] at 60 minutes of reperfusion (P<0.05, vs control). Partial correlation analysis revealed a negative correlation of[Ca2+]I with insulin-induced ghcose uptake in the cardiomyoctyes (r = -0.557,P=0.006). Conclusion Disturbed GLUT4 translocation and decreased intrinsic activity may be important molecular mechanisms for the development of insulin resistance in the cardiomyocytes of rat during insulin-simulated ischemia reperfusion,. [Ca2+]I overload may account for the de- creased intrinsic activity d GLUT4.

Objective To investigate the expression of sodium iodide symporter (NIS ) mRNA in differentiated thyroid Carcinoma (DTC)and further explore its value in clinical diagnosis and therapy of DTC. Methods The expression of NIS mRNA was detected and analyzed in 21 nodular goiter and 45 cases of DTC (including 35 cases of papillary thyroid carcinoma and 10 cases of follicular thyroid carcinoma)by using real-time fluorescent quantitative reverse transcription polymerase chain reaction (real-time RT-PCR).Results Compared with that in nodular goiter,the mRNA expression of NIS in DTC tissue was significantly decreased (P < 0.05 ). Moreover,the mRNA expression of NIS was significantly correlated with lymph node metastasis and AJCC stage, respectively.The expression of NIS mRNA in DTC with lymph node metastasis was significantly decreased compared with that in DTC without lymph node metastasis (P <0.05).In addition,the expression of NIS mRNA in Ⅱ-Ⅳstage DTC was significantly decreased compared with that in Ⅰ - Ⅱ stage DTC (P < 0.05 ).Conclusion Differential expression of NIS can provide evidence for individual 1 3 1 I therapy for DTC.

Objective To evaluate the role of neuropeptide Y2 receptor (NPY2R) in neuropathic pain (NP) in rats.Methods Thirty-six adult male Sprague-Dawley rats,aged 8 weeks,weighing 190-210 g,were randomly divided into 3 groups (n =12 each):sham operation group (group S),group NP and NPY2R antisense ohgonucleotide group (group ODN).NP was induced by chronic constrictive injury (CCI).5 μg/μl NPY2R antisense oligonucleotide 30 μl was injected intrathecally 7 days after CCI in group ODN.While normal saline 30 μl was injected intrathecally in group S.The mechanical paw withdrawal threshold and cold allodynia were measured 3 days before CCI (T0,baseline),7 days after CCI (T1) and at 15 min,1.5,3.0,4.5 and 6.0 h after intrathecal injection (T2-6).The animals were then sacrificed after the last measurement and the lumbar segment of spinal cord was removed for determination of the expression of NPY2R and calcitonin gene-related peptide (CGRP) and co-expression of NPY2R with CGRP in spinal dorsal horn neurons (by immuno fluoresceence).Results Compared with group S,the mechanical paw withdrawal threshold was significantly decreased and cold allodynia was increased at T1-6,and the expression of NPY2R and CGRP and co-expression of NPY2R with CGRP in spinal dorsal horn neurons was up-regulated in NP and ODN groups (P ＜ 0.05).Compared with group NP,the mechanical paw with-drawal threshold was significantly increased at T3-5,and the expression of NPY2R and co-expression of NPY2R with CGRP in spinal dorsal horn neurons was down-regulated (P ＜ 0.05),and no significant change was found in cold allodynia and the expression of CGRP in spinal dorsal horn neurons in ODN group (P ＞ 0.05).Conclusion NPY2R in the spinal cord dorsal horn is involved in the maintenance of mechanical hyperalgesia,but not in the maintenance of clod hyperalgesia in rats.

Objective To investigate the galectin-3 expression in squamous carcinioma of esophagus and analyze its relationship with clinicopathological characteristics.Methods S-P immunohistochemistry was used to detect the expression of galectin-3 in 80 paraffin-embedded specimens of esophageal carcinoma and 22 specimens of normal tissues adjacent to the benigh lesions of esophagus.RT-PCR was performed to detect galectin-3 mRNA expression in 36 pairs of fresh squamous carcinoma tissues and corresponding adjacent normal mucosa.Results Out of 80 specimens of squamous carcinoma of esophagus,62 specimens had positive expression of galectin-3,with the positive rate of 77.5%(62/80)that was significantly higher than 27.27% in normal esophageal tissues(6/22,P

Objective To explore the indication and the functional advantages of the high-flexion posterior stabilized (PS) rotating-platform mobile-bearing (RP-MB) total knee system. Methods A prospective randomized, controlled trial was performed. Osteoarthritis was the indicators for total knee arthroplasty.From Feb. 2009 to Apr. 2009, 75 patients (94 knees) were randomly assigned to to receive either a highflexion PS, RP-MB total knee system(PFC sigma RPF) or a standard one (PFC sigma RP). There were no statistical difference in the baselines, the preoperative scores of the Hospital for Special Surgery (HSS) and the knee range of motion (ROM) of both groups. The functional status were assessed with Hospital for Special Surgery and the ROM of the knee at the postoperative 1, 6, 12, 18 months. The satisfaction rates were assessed at the postoperative 18 months. The radiographic measurements were t assessed at the postoperative 3days and 3, 6, 12, 18 months. Results A total of 70 participants (87 knees) completed the 18-month followup. At the time of the final follow-up, the average Hospital for Special Surgery knee score was 92.4±5.0points in the standard group and 94.7±7.0 points in the high-flex ion group. The difference was not statistically significant(P ＞0.05). The average maximal flexion was 131.9±14 degrees in the high-flexion group and 123.0±15.3 degrees in the standard group. There was a statistical difference. But it was not enough to confirm our hypothesis that the difference should be higher than 10 degrees. Moreover, the satisfaction rate were 100% in both groups, and no statistical significant difference was found. Conclusion No significant differences were found between standard and high-flexion posterior-stabilized rotating-platform mobile-bearing total knee prostheses in terms of clinical outcomes or range of motion.

ObjectiveTo explore the association between hepatic steatosis and the liver tissue expression of HBsAg and HBcAg in chronic hepatitis B (CHB) patients.MethodsFrom January 2005 to June 2008,a total of 147 CHB patients with hepatic steatosis diagnosed by liver biopsy and other 149 CHB patients without hepatic steatosis but with similar HBV DNA titer were enrolled.The differences of HBsAg and HBcAg immunostaining and liver injury in these two groups were compared.The data were analysed using t test and chi square test.ResultsCompared with non-steatosis group,the average age and body weight index of hepatic steatosis group were higher (t values were -3.31and -6.57,both P＜0.01).The percentage of moderate to severe hepatic inflammation in liver,obvious hepatic fibrosis and the strong positive HBsAg staining was lower (30.6％ vs 15.4％ ； 26.5％vs 12.8％； 23.1 ％ vs 6.7 ％； x2=9.63,8.92,15.76； all P＜0.01),and the percentage of strong positive HBcAg staining was also in downtrend.Compared with degree F1 and F2 of liver steatosis,the percentage of HBsAg and HBcAg strong positive staining in liver tissues of degree F3 and F4 was in downtrend.ConclusionsHepatic steatosis affected the expression of HBsAg and HBcAg in liver tissue of CHB patients.As hepatic steatosis appeared and became more severe,both expression of HBsAg and HBcAg and the degree of liver injury were in downtrend.

Objective To investigate the correlation between ICRU reference point dose and dosevolume parameters of organs at risk (OARs) under different bladder and rectal filling status in threedimensional conformal brachytherapy for locally advanced cervical cancer.Methods A total of 31 patients who received magnetic resonance imaging-guided three-dimensional conformal brachytherapy for cervical cancer in 96 fractions were enrolled.The ICRU rectal and bladder reference points were determined in the treatment planning system,and the doses at these points were recorded and compared with the dose-volume parameters of the rectum and bladder.The paired t-test was used to analyze the differences between them.Results Bladder DICRU was lower than bladder D0.1cm3 and D1 cm3 (P=0.000 and 0.000),higher than bladder D5 cm3 and D10cm3 (P=0.000 and 0.000),and similar to bladder D2 cm3 (P=0.345).Under the bladder filling status,bladder DICRU was lower than D2cm3.Rectal DICRU was lower than rectal D0.1 cm3 and D1cm3 (P =0.000 and 0.002),higher than rectal D5 cm3 and D 10 cm3 (P =0.000 and 0.000),and similar to rectal D2cm3 (P=0.058).The ICRU bladder and rectal reference point doses were positively correlated with corresponding D2 cm3.In the case of bladder volume ≥ 200 cm3,the ICRU bladder reference point dose underestimated bladder D2 cm3.In the case of rectal volume ≥ 37 cm3,the ICRU rectal reference point dose overestimated rectal D2 cm3.Conclusions In three-dimensional conformal brachytherapy,it is generally safe to use D2 cm3 as an index to evaluate OARs,but when the bladder or rectum is in an empty status,the ICRU bladder or rectal reference point doses should be considered.

Objective To explore clinical feasibility of liver transplant from child of brain death to adult, to summarize the clinical experiences that a child of brain death transplants liver to an adult. Methods The recipient was a 39-year-old woman patient with primary hepatic carcinoma and posthepatitis cirrhosis (decompensation stage); while the donor was a 8-old-year child of brain death because of brain neoplasms. Donated liver was gained by the method of en bloc multivisceral procurement in a short time; the operative method was classic orthotopic liver transplantation. The postoperative managements included immunosuppression, prevention of infection, hepatic protection, and other relevant supports etc. Results The transplantation operative duration was 6 hours, after which not only did the recipient survive but also her body functioned well including the liver part, with no severe postoperative complications. Conclusions The technology of transplanting livers from children to adults is feasible. The key to ensure the success of transplant operation is systematic preoperative evaluation, excellent operative technique, and perfect postoperative treatment.

BACKGROUND: Previous studies demonstrated that construction of eoexpression plasmid containing multiple genes on the same vector could improve transfection and expression rates.OBJECTIVE: To construct eukaryotic expression plasmid pcDNA3.1 (+)-MUC1 -GM-CSF by human polymorphic epithclial mucin (MUC 1) and granulocyte macrophage colony stimulating factor.(GM-CSF) and to observe expression of recombinant plasmid in COS-7 cells.DESIGN,TIME AND SETTING: Gene recombination design,which was carried out in the Animal Central Laboratory,the Fourth Military University of Chinese PLA from September 2005 to December 2006.MATERIALS: Eukaryotic expression vector pcDNA3.1 (+) was presented by Pro.Taylor-Papadimitriou;pGEM-3zf()-GM-CSF plasmid,COS-7 cells,pUCI 8 vector,and E.coli DH5α were made in the center; female BALB/c mice were provided by Experimental Animal Center of the Fourth Military University of Chinese PLA.METHODS: Signal peptide was synthesized with encoded MUCI gene sections to obtain repeated sequence coneatemer after renaturation.Next,the accepted concatemer was cloned with GM-CSF following enzyme identification and sequencing analysis,and then they were put in eukaryotic expression vector pcDNA3.1(+) to construct eukaryotic coexpression plasmid pcDNA3.1 (+)-MUCI -GM-CSF in order to transform COS-7 cells.MAIN OUTCOME MEASURES: Gene expression was detected by indirect immunofluorescence and enzyme linked immunosorbent assay (ELISA).RESULTS: Enzyme identification and sequencing analysis showed that recombinant plasmid contained a fusion gene encompassing human MUC1 repeated sequence concatemer and GM-CSF; moreover,MUC1 expression was detected in COS-7 cells,while recombinant plasmid could induce the production of anti-GM-CSF antibody.CONCLUSION: The recombination between human MUC1 repeated sequence concatemer and GM-CSF gene successfully constructs eukaryotic coexpression plasmid.