Objective　To study the expression of 5 multidrug-resistant genes in hepatocellular carcinoma (HCC) and establish the criteria for genetic diagnosis of multidrug resistance (MDR). Methods　The expression of mRNA and proteins of the multidrug-resistant genes were detected with reverse transcriptase polymerase chain reaction (RT-PCR) and flow cytometry (FC) respectively. IC50 of the original HCC cells was determined with MTT method. Results　MDR in HCC was associated with the genes of mdr1, MRP, LRP, Topo II α and GSTTP1. mdr1 mRNA ≥0.5, MRP mRNA ≥0.6, LRP mRNA ≥0.8, GSTP1 ≥0.7 and Topo II α mRNA ≤0.4 could be used as the criteria for genetic diagnosis of MDR in HCC. Conclusion　The complex MDR is the main form of drug resistance of HCC. Using RT-PCR to detect the mRNA expression of 5 multidrug-resistant genes is necessary and feasible for predicting sensitivity of chemotherapy for HCC.

Objective: To analyze clinical features and prognosis of more than 75-year old patients with heart failure and preserved ejection fraction (HFPEF) through comparing with heart failure（HF）and reduced ejection fraction (HFREF), and provide reference for prevention and treatment of HFPEF. Methods: A total of 134 > 75 years HF inpatients admitted from Jan 2009 to Dec 2011 were selected. With left ventricular ejection fraction (LVEF) 50% as the critical point, patients were divided into HFPER group (n=63) and HFREF group (n=71). Clinical characteristics were compared between two groups. Clinical outcomes (all-cause death and rehospitalization caused by heart failure) were compared between two groups after follow up. Results: (1) Clinical features: compared with HFREF group, there were significant increase in LVEF [(36.46±6.84)% vs. (58.65±5.01)%], percentage of patients with hypertension (49.3% vs. 69.8%), left atrial diameter [(34.98±3.78) mm vs. (40.02±3.29) mm], and significant decrease in level of brain natriuretic peptide [BNP, (1005.62±458.99) pg/ml vs. (646.57±333.56) pg/ml], concentration of hemoglobin [(11.97±1.29) g/dl vs. (10.76±1.21) g/dl] and left ventricular diameter [(57.17±7.52) mm vs. (47.73±5.48) mm] in HFPEF group, P<0.01 all; (2) Patients were followed up for a mean 2.6 years. There were no significant difference in all-cause mortalities（17.5% vs. 19.7%）and mean time without heart failure event (286 d vs. 258 d) between HFPEF group and HFREF group. Conclusion: Compared with HF patients with reduced ejection fraction, hypertension and anemia are more frequent in HF patients with preserved ejection fraction, but there is no significant difference in clinical outcomes between them.

Aim To investigate the effect of tanshinol on bone mineral density and microstructure of proximal tibias in rats with bone loss induced by glucocorticoid. Methods Sixty 7-month-old female SPF SD rats were randomly divided into 6 groups with 1 0 rats per group:control group(saline:5 ml·kg -1 ·d -1 ),glucocorti-coid group (prednisone acetate:6 mg·kg -1 ·d -1 ), glucocorticoid +low dose of tanshinol group(1 2.5 mg ·kg -1 ·d -1 ),glucocorticoid +medium dose of tan-shinol group (25 mg·kg -1 ·d -1 ),glucocorticoid +high dose of tanshinol group (50 mg·kg -1 ·d -1 ), glucocorticoid +(positive control drug)calcitriol group (0.045 μg · kg -1 · d -1 ).Rats were gavaged with prednisone acetate continuously for 1 4 weeks to estab-lish the bone loss model.Meanwhile,tanshinol and calcitriol were orally administered to the rats which were treated with prednisone acetate for intervention. At the end of the experiment,the left proximal tibias were collected for Micro-CT scanning and three-dimen-sional reconstruction of cortical and trabecular bone re-
spectively to observe the changes of bone microstruc-ture and test related parameters.Results Bone min-eral density was decreased and bone microstructure was destroyed in proximal tibias of rats after treatment with glucocorticoid.Both tanshinol (25 mg·kg -1 ·d -1 ) and calcitriol(0.045 μg·kg -1 ·d -1 )could increase bone mineral density and improve bone microstructure in proximal tibias without significant differences be-tween each other.Tanshinol (50 mg · kg -1 · d -1 ) could improve bone microstructure to some extent,but it had no significant effect on bone mineral density. Tanshinol(1 2.5 mg·kg -1 ·d -1 )had no significant effect on bone mineral density or microstructure.Con-clusion Oral administration of tanshinol (25 mg · kg -1 ·d -1 )to the rats treated with glucocorticoid can increase bone mineral density and improve bone micro-structure in proximal tibias.

Objective:To evaluate the effect of Arg and Gln enriched PN on nutrition status,immune function and stress status of gastric cancer patients after radical treatment.Methods:Eighty patients with gastric cancer were randomly divided into 4 groups: arg group(arginine enriched PN group),Gln group(glntamine supplemented PN group),Arg+Gln group(arginine and glutamine enriched PN group)and Rou group(Routine PN group).The distinct immune nutrients were added into PN in the different groups for 7 consecutive days.The difference of the patients' nutrition status,immune function and stress status were analyzed after operation.Results: The nutritional station and immunue fuction in Arg group,Gln group and Arg+Gln group were better than in Rou group on the 8th postoperative day.The concentration of CRP in the Arg+Gln group was different from that in other groups(P

Objective To analysis curative effect of triple therapy and sequential therapy for eradication of Helicobacter pylori in patients with long-term aspirin use.Methods 52 patients with long-term aspirin use who were diagnosed with helicobacter pylori infection were collected.All patients were divided into sequential group and triple group according to different drugs classified, each 26 cases in each group were given corresponding drug treatment, after the end of treatment, the serum pepsinogen, Hp-IgG antibody levels and Helicobacter pylori clearance rate were detected in all patients. Results After treatment, compared with the triple Group, in the sequential group, the levels of serum PG I and PG II were lower, and PGⅠ/PGⅡwas higher, and the differences were statistically significant (P<0.05);the serum Hp-IgG antibody levels were lower in the sequential group (P<0.05);the Hp clearance rate of patients in the sequential group was higher (P<0.05).Conclusion Compared with triple therapy, sequential therapy can lower serum PG, PG II and Hp-IgG levels, and improve the Hp clearance rate and clinical efficacy, have guiding significance to clinical.

Objective To investigate the effects of sericin pretreatment on the expression of extracellular matrix(ECM) associated protein in diabetic nephropathy(DN) rats' kidney.Methods Sixty six male SD rats were randomly divided into 3 groups(n=12):normal control group,DN model group and sericine pretreatment group.DN rats model in model group and sericine pretreatment group were established by intraperitoneally injection of streptozotocin(STZ).Blood glucose≥16.7 mmol/L was taken as the standard of successful modelization.The rats in sericine pretreatment group were lavaged with sericine(2.4 g·kg~(-1)·d~(-1)) for 35 days before injecting STZ.The enzymic method was used to measure the blood glucose.Type Ⅳ collagen(cⅣ)and laminin(LN)content in the serum were detected by ELBA.The expression of transforming growth factor-β_1,(TGF-β_1)and tissue inhibitors of maprix metalloproteinase-1(TMP-1) protein in the kidney was observed by immunohistochemical staining.The expression of Smad 3 protein in the kidney was detected by Western blot.Results Compared with normal control rats,the blood glucose,cⅣ and LN content in the serum,TGF-β_1,TIMP-1 and Smad 3 expression in the kidney of the model group rats increased obviously(P<0.01).The blood glucose,cⅣ and LN content in the serum,TGF-β_1,TMP-1 and Smad3 expression in the kidney of rats in sericine pretreatment group were significantly lower than those of the rats in model group(P<0.01).Conclusion Sericin pretreatment can inhibit the activation of TGF-β/Smad 3 signal pathway in the kidney of DN rats,and prevent the decrease of MMPs activity induced by up-regulation of TIMP-1.So sericin can prevent accumulation of ECM and glomerulosclerosis during DN,and has satisfactory apotropaic effects on the development of DN.

In the present research, the effects of sintered bone modified with surface mineralization/P24 peptide composite biomaterials on the adhesion, proliferation and osteodifferentiation of MC3T3-E1 cells were investigated. The experiments were divided into three groups due to biomaterials used: Group A (composite materials of sintered bone modified with surface mineralization and P24, a peptide of bone morphogenetic protein-2); Group B (sintered bone modified with surface mineralization) and Group C (sintered bone only). The three groups were observed by scanning electron microscopy (SEM) before the experiments, respectively. Then MC3T3-E1 cells were cultured on the surfaces of the three kinds of material, respectively. The cell adhesion rate was assessed by precipitation method. The proliferative ability of MC3T3-E1 cells were measured with MTT assay. And the ALP staining and measurement of alkaline phosphatase (ALP) activity were performed to assess the differentiation of cells into osteoblasts. The SEM results showed that the materials in the three groups retained the natural pore structure and the pore sizes were in the range between 200-850 μm. The adhesive ratio measurements and MTT assay suggested that adhesion and proliferation of MC3T3-E1 cells in Group A were much higher than those in Group B and Group C (P < 0.05). The ALP staining and ALP activity of MC3T3-E1 cells in Group A were significantly higher than those in Group B and Group C (P < 0.05). The sintered bone modified with surface mineralization/P24 composite material was confirmed to improve the adhesion rate and proliferation and osteodifferentiation of MC3T3-E1 cells, and maintained their morphology.
3T3 Cells ; Animals ; Biocompatible Materials ; Bone Morphogenetic Protein 2 ; Bone and Bones ; Cell Adhesion ; Cell Differentiation ; Mice ; Microscopy, Electron, Scanning ; Osteoblasts ; cytology ; Peptides ; Staining and Labeling

Objective To explore the effects of combined infusion of Treg cells from recipient rats and imDCs from donor rats prolonging the survival time of rats undergoing allogeneic orthotopic liver transplantation.Methods One hundred cases of rat allogeneic orthotopic liver transplantation were performed by using modified two-cuff method: DA rats served as donors,and Lewis rats as recipients,which were divided into 5 groups: acute rejection group,mDC group,imDC group,Treg group,co-infusion group (imDC-Treg group).2 × 106 imDC,2 × 106 mDC and 1 × 107 Treg cells were respectively injected to Lewis rats in every group via caudal vein before transplantation.Four Lewis rats from each group were randomly sacrificed on the day 3,7 and 10 after transplantation.Liver function (ALT and TBIL) and changes of cytokines (IL-10,IL-2 and TGF-β) were evaluated in peripheral blood.The pathological changes of the liver graft were observed.The DNA fragmentation characteristics of apoptosis in the liver were detected by using TUNEL staining.Survival analysis was performed on remnant rats.Results The liver function in imDC-Treg group had reverted to normal faster than that in the control groups.ELISA revealed that at day 7,the serum levels of IL-10 and TGF-β were higher,and the serum levels of IL-2 were lower in imDC-Treg group.Liver pathology demonstrated that there was only a slight number of mononuclear cell infiltration in hepatic graft afterorthotopic liver transplantation in co-infusion group (imDC-Treg group),which belonged to a nonidentified or slight type of acute rejection,and the liver graft showed regeneration of hepatocytes along with a small number of neutrophile granulocyte infiltration at the day 10.TUNEL staining indicated that the number of apoptotic T cells was much less in control groups than in imDC-Treg groups.The survival time of recipients in Treg-imDC group was 37 days,significantly longer than in irnDC group (23 days) and Treg group (15 days).Conclusion Infusion of imDC from donor rats or Treg from recipient rats alone couldn't induce long-term allograft survival of recipients after allogeneic rat liver transplantation.The co-infusion of Treg and imDC significantly prolonged the survival time of rats subject to allogeneic liver transplantation.

Objective:To establish the optimal extraction technology of Xiaoer Yinqiao granules by orthogonal test. Methods:The effects of water amount,extraction duration and extraction times were investigated by orthogonal design using the contents of forsythia-side A and chlorogenic acid as the indices. Results: The optimum extraction process was as follows: adding 8-fold amount of water, and extracting 1. 5 h for the first time, and then adding 6-fold amount of water, extracting 1 h for the second and third time, respective-ly. Conclusion:The extraction technology is simple, reasonable and reliable.

Objective To investigate the effects of radix salviae miltiorrhiaze(RSM) on the expression of Fas, perforin and bcl 2 mRNA in liver ischemia reperfusion(I R). Methods 45 patients were divided into three groups: Control group(group 1, n=5); I R group(group 2, n=20), and RSM pretreatment group(group 3, n=20). The patients in group 2 and group 3 subjected to hepatic resection with “Pringle procedure” for 15?min. Expression of Fas, perforin and bcl 2 mRNA were examined with reverse transcription polymerase chain reaction(RT PCR) after liver reperfusion for 40?min. Results Expression of Fas, perforin were very weak in control group(0.144?0.037; 0.106?0.032), but increased in group 2(0.928?0.135; 1.099?0.364); and in group 3 was significantly lower(0.347?0.081; 0.454?0.116)than that in group 2(P