AIM:To investigate the expression of long non-coding RNA maternally expressed gene 3 (MEG3) in colorectal cancer ( CRC) cells, and to observe the effect of MEG 3 on the invasion and migration of CRC cells .METH-ODS:The levels of MEG3 in human normal colon cell NCM 460 and CRC cells SW48 and LoVo were detected by real-time PCR.MEG3 was over-expressed by plasmid transfection , and the effects of MEG 3 on the invasion and migration of SW 48 and LoVo cells were analyzed by Transwell assay and wound healing assay .The expression of matrix metalloproteinase ( MMP) family proteins was determined by Western blotting .RESULTS:The level of MEG3 was down-regulated in CRC cells compared with normal colon cell NCM 460.The invasion and migration of CRC cells were reduced after MEG 3 over-ex-pression.Transwell invasion and migration assays showed that the numbers of transmembrane SW 48 and LoVo cells were smaller in MEG3 over-expression group than control group (P<0.05).The cell spaces were broader after MEG3 over-ex-pression in the wound healing assay , indicating that MEG3 over-expression inhibited the mobility of CRC cells .Meanwhile, over-expression of MEG3 reduced the expression of MMP-2 and MMP-9, and elevated the expression of tissue inhibitor of metalloproteinase-2 (TIMP-2).CONCLUSION:The expression of MEG3 is down-regulated in CRC cells.Over-expres-sion of MEG3 inhibits the invasion and migration of CRC cells .TIMP-2, MMP-2 and MMP-9 might play an important role in this regulation .

ObjectiveTo investigate the effect of pravastatin on serum levels of interleukin-6(IL-6) and tu- mor necrosis faetor-α(TNF-α) in patients with acute coronary syndromes(ACS). Methods50 patients with ACSwere randomly divided into pravastatin group( n = 25) and routine therapy group( n = 25). Serum TNF-α and IL-6levels were measured before and four weeks after the two treatment options respectively. ResultsThe level of TNF- α and IL-6 were higher than routine therapy group before therapy(P < 0.01 ). The level of TNF-α and IL-6 weredecreased significantly after pravastatin therapy,and higher thancontrols(P < 0.05). The levels of TNF-α and IL-6changed only slightly after the routine therapy(P > 0.05). ConclusionThe level of TNF-α and IL-6 becominghigher in ACS patients may be related to the pathogenesis of ACS. Pravastatin can reduce serum level of TNF-α and IL-6 contributing to treatment of ACS.

Objective To evaluate the effects of continuous quality improvement (CQI) management on nutritional status,renal function progression,and compliance of low protein diet in patients with chronic kidney disease (CKD).Methods Totally 115 CKD patients who were regularly followed up in CKD clinic services were recruited in this study.Plan,Do,Check,and Act (PDCA) method was adopted to manage the dietary of these patients for 12 months.The clinical indicators and diet compliance before and after receiving CQI management were compared.Results After receiving the CQI management,the nutritional status of patients was well maintained;meanwhile,the average hand strength and the hemoglobin,serum albumin,total cholesterol,and triglyceride levels showed no significant changes (all P ＞ 0.05).Subjective feelings of patients were improved.The modified Subjective Global Assessment of Nutrition (mSGA) score was decreased from 7.0 (7.0,8.0) to 7.0 (7.0,7.0) (P =0.000).The estimated glomerular filtration rate (eGFR) calculated by formula of modified MDRD was decreased from (40.74 ± 14.49) to (37.94 ± 16.86) ml/(min · 1.73 m2) (P =0.000),and the average descended speed was (2.81 ±7.42) ml/(min · 1.73 m2) per year.The creatinine clearance rate had no statistical difference between pre-and post management (P =0.910),and the average descended speed was (0.19 ± 17.01) ml/min per year.The daily protein intake (DPI) and protein equivalent of nitrogen appearance rate (PNA) were both significandy descended:DPI/kg decreased from (0.79 ± 0.27) to (0.64 ± 0.15) g/ (24 h · kg) (P =0.000),and PNA/kg dropped from (1.02 ± 0.32) to (0.82 ± 0.24) g/ (24 h · kg) (P =0.000).The scores of awareness and compliance of patients on low protein diet were significantly increased after CQI management (P =0.000).Conculsion Applying CQI on dietary and nutrition management in CKD patients can maintain the good nutritional status and improve the compliance of low protein diet.

Objective To study the effect of long-term salmon calcitonin on bone mineral density (BMD), bone metabolism biochemical indicators and subjective score of bone pain in maintenance hemedialysis (MHD) patients with osteopenia. Methods Thirty-four MHD patients diagnosed as osteopenia by dual-energy X-ray absorptiometry (DXEA) were enrolled in this study. All the patients were treated with hypodermic injection of salmon calcitonin (50 U, thrice a week) for 12 months. The detecting parameters were as follows: BMD with DEXA in lumbar spine (L1-L4), femoral neck, troch, inter, and Ward's triangle before and after the study;serum bone metabolism biochemical indicators before and 6 and 12 months after the study;subjective scores of bone pain before and 1, 6, and 12 months after the study. Results Thirty-two patients were followed-up successfully. As compared to BMD parameters before study, the total T-score (-1.98± 2.20 vs 1.26±1.88, P=0.009) and total Z-score (-0.90±2.15 vs 0.08±2.05, P=0.002) of lumbar spine, the total T-score (-1.72±1.53 vs 1.06±1.58, P=0.016) and totle Z-score (-0.66±0.80 vs 0.08±1.08, P=0.029) of hip, the T-score of L3 (-2.02±2.51 vs 1.24±2.02, P=0.033), the Z-score of L2 (-0.44±1.82 vs 0.06±1.63, P=0.016), the Z-score of femoral troch (-0.65±1.11 vs 0.48±1.12, P=0.034) and the Z-score of inter (-0.58±0.94 vs 0.02±1.12, P=0.006) were increased significantly after study. But there were no significant differences in other examined regions and serum biochemical parameters. The subjective scores of bone pain were decreased rapidly for 41.7% after 1 month (P<0.01) and 76.6% after 6 months (P<0.01). The subjective score of bone pain after 12 months was similar to 6 months. The side effects of salmon calcitonin included nausea and vomitting in 5 cases (14.71%, 5/34), dizziness, blushing and flustered in 1 case respectively (3.13%,1/32). Conclusions Long-term hypodermic injection of salmon calcitonin can improve BMD and bone pain for MHD patients with osteopenia but has no significant effect on serum bone metabolism biochemical indicators. Salmon calcitonin is safe for MHD patients with seldom side effects, such as nausea and vomitting.

AIM:To investigate the formation of membrane pore in PC 12 cells induced by exogenous adenosine triphosphate ( ATP) and to identify the key molecular targets .METHODS:PC12 cells were treated with different concen-trations of ATP to establish the injury model .The morphological change was observed under an inverted phase -contrast mi-croscope.The viability of the PC12 cells was measured by CCK-8 assay.Fluorescent dye YO-PRO-1 was used to detect the membrane permeability.The expression of P2X7 receptor and pannexin 1 (Panx1) at mRNA and protein levels was as-sessed by real-time PCR and Western blotting .RESULTS:After exposed to ATP (1 mmol/L, 3 mmol/L and 5 mmol/L) for 3 h, the PC12 cells became edematous , and the number of adherent cells decreased gradually in a dose-dependent man-ner .The cell viabilities in 3 mmol/L ATP group and 5 mmol/L ATP group were significantly decreased compared with con-trol group (P<0.05).YO-PRO-1 uptake in the PC12 cells exposed to ATP (0, 1, 3 and 5 mmol/L) for 15 min, 30 min and 60 min increased in a dose-dependent and time-dependent manner .The cell viability increased and the intracellular fluorescence intensity induced by ATP were significantly antagonized in brilliant blue G ( a P2X7 receptor inhibitor ) pre-treatment group (P<0.05), whereas it did not change in carbenoxolone (a Panx1 inhibitor) pretreatment group (P>0. 05).The expression of P2X7 receptor at mRNA and protein levels was significantly increased (P<0.05), but the expres-sion of Panx1 was not changed ( P>0.05) when PC12 cells were exposed to ATP for 3 h.CONCLUSION:Extracellular ATP at high concentration may induce membrane pore formation with the expression and activation of P 2X7 receptor in PC12 cells.

Objective To clarify the clinical significance of visit-to-visit variability in blood pressure (BP) of stage 3-4 chronic kidney disease (CKD) patients with hypertension.Methods One hundred and fifty-two cases of stage 3-4 CKD patients with hypertension were enrolled in the study.Variability in BP was defined as the standard deviation (SD) in BP.For each patient,SD and mean BP from BP measurements were calculated at all the visits.Correlations between the decline in estimated glomerular filtration rate (eGFR) and SD in BP were analyzed by multivariable regression.Results Visit-to-visit variability in BP was significantly associated with renal function decline (P ＜ 0.05),in addition,baseline eGFR,baseline albuminuria and mean SBP during follow-up were significantly associated with renal function decline as well (all P ＜ 0.05).The percentage of CCBs used in low SD of the SBP group was higher than that in high SD of the SBP (76.1％ vs 58.2％,P ＜ 0.05).Conclusion Visit-to-visit variability in BP is significantly associated with renal function decline.Drugs which can decrease the variability of blood pressure should be the first choice in the treatment of hypertension.

BACKGROUND: Adequate preparation of donors and recipients prior to living-related donor renal transplantation, short warm and cold ischemia time for donor kidney, good histocompatibility of human leukocyte antigen match, and low postoperative rejection incidence provide feasibility for use of low-dose immunosuppressive agents after living-related donor renal transplantation. OBJECTIVE: To investigate the safety and effectiveness of low-dose calcineurin inhibitors (CNI), an immunosuppressive agent, in living-related donor renal transplantation. METHODS: A total of 38 recipients who underwent living-related donor renal transplantation at the Center of Renal Transplantation of the First Affiliated Hospital of Nanjing Medical University from January 2006 to June 2008 were randomized for treatment with mycophenolate mofetil (750 mg twice a day), prednisone, and either standard-dose CNI (n=18) or low-dose CNI (n=20) during 12 months post-transplantation. Ciclosporin A was given orally (starting dose, 6 and 4 mg/kg per day, respectively) in two divided doses to achieve the 12-hour whole blood concentration as measured by fluorescence polarization immunoassay. The starting dose of tacrolimus was 0.12 and 0.08 mg/kg per day respectively, and its whole blood concentration was measured by enzyme-multiplied immunoassay technique. After transplantation, patients were followed up. Renal function, pulmonary infection, liver dysfunction, and CNI nephrotoxicity at different time periods were compared between different regimens. RESULTS AND CONCLUSION: During 12 months post-transplantation, patient death occurred in one of 18 patients (5.6%) in the CNI standard-dose group and none of 20 patients (0%) in the CNI low-dose group. There was no significant difference in renal function and acute rejection between CNI standard-dose and CNI low-dose groups (P > 0.05). The incidence of liver dysfunction and CNI nephrotoxicity was significantly lower in the CNI low-dose group than in the CNI standard-dose group (P < 0.05). In addition, a low-dose CNI regimen helped recipients to lessen the economic burdens. These findings indicate that it is effective, safe and economical to use a low-dose CNI regimen in living-related donor renal transplantation.

Objective To investigate the expression of OPN, α-SMA, E-cadherin and their correlation in the chronic allograft nephropathy (CAN) rat model, and to explore the possible role of OPN in CAN.Methods Orthotopic renal-transplantation using Fisher rats as donors and Lewis rats as recipients was done to establish CAN group, and Lewis to Lewis rats as control group. Rats in each group were sacrificed 12 weeks after the surgery. Blood and urine were collected for further test. Allograft samples were collected and sectioned for HE, Sirus-red staining, immunohistochemistry and Western blot.Results There were CAN morphological changes of the allograft in CAN group. As compared with control group, immunohistochemistry and Western blot revealed that the expression of OPN and α-SMA in CAN group was significantly increased, and that of E-Cadherin reduced. Its trend was correlated with the inflammatory response and the EMT of tubule epithelial cells.Conclusions OPN expression in rat CAN model is significantly up-regulated. OPN may play a role in CAN. OPN might affect the CAN by promoting EMT of tubule epithelial cells.

Objective To evaluate the protective effect of adiponectin in early atherosclerosis and the diagnostic value of adiponectin in metabolic syndrome in obese children. Methods Total 176 obese children and 88 normal weight children aged 9-12 years were included in the present study. All participants underwent hematologic and biochemical tests including serum adiponectin, high sensitivity C-reactive protein (hsCRP),fasting blood glucose, insulin, and plasma lipids. Homeostasis model assessment of insulin resistance (HOMA-IR)was calculated. Noninvasive ultrasound measurement including intima-media thickness of the common carotid artery(IMT), brachial flow-mediated dilatation (FMD), carotid artery compliance (CAC), and the maximum fatthickness ahead of peritoneum (Pmax) were obtained to investigate arterial mechanical properties and endothelial function. Results (1) The level of adiponectin was negatively correlated with obese index, blood pressure,fasting insulin, hsCRP, HOMA-IR, and IMT(P<0.05 or P<0. 01 ); but not with triglyceride, fasting blood glucose, CAC, high-density lipoprotein-cholesterol (HDL-C), and FMD. (2) The risk of metabolic syndrome increased 3.43 times in children with adiponectin level <7. 060 mg/L compared with ＞7. 060 mg/L. (3)Receiver operating characteristic( ROC ) curve was used to choose the optimal cutpoint of adiponectin to identify obese children with the metabolic syndrome. The area under the curve (AUC) for adiponectin to discriminate the sensitivity of metabolic syndrome was 0. 769 (95% CI0. 714-0.816, P< 0. 0 1 ). (4) The obese children were divided into three groups according to the cut-off value for adiponectin (high, middle, low groups). There were significant differences in the prevalences of severe obesity, visceral fat accumulation, hypertension, insulinemia,low HDL-C, metabolic syndrome among three groups (P<0.05). Conclusions High levels of serum adiponectin could prevent early stage of atherosclerosis. The lower the adiponectin level, the higher the incidence of metabolic syndrome.

Objective To study the feasibility of artificial immune system(AIS)in assistant designing of computer for new Chinese herbal compounds with anti-infectious function.Methods A group of Chinese herbal drug compounds from Treatise on Exogenous Febrile and Miscellaneous Diseases were selected as training samples.Serum was obtained from mice which were infected by Staphylococcus aureus and ig administrated with the various Chinese herbal compounds.The inhibition ability against S.aureus of the serum was tested and input into an artificial neural network(ANN)together with the characters of the every Chinese herb included in those compounds.The ANN was optimized using AIS so that it can predict the inhibitory ability against S.aureus of the Chinese herbal compounds according to the characters of Chinese herbs included in those compounds.After that the computer generated a number of Chinese herbal compounds randomly,calculated their inhibitory ability against S.aureus and output those among the top levels,so as to verify whether the predictions were correct or not.Results After verification,the Chinese herbal drug compounds designed by computer could significantly raise the serum's inhibitory ability against S.aureus.Conclusion The computer can be an assistant for designing new Chinese herbal compounds with anti-infectious function