Objective: To observe the effects of knocking down the expression of peroxisome proliferators-activated receptor gamma (PPARγ) with RNA interference techniques on the proliferation and apoptosis of hepatocellular carcinoma HCCLM3 cells. Methods: A short-hairpin RNA (shRNA) eukaryotic expression vector against PPARγ was constructed and transfected into HCCLM3 cells. The changes of PPARγ expression were detected by semi-quantitative RT-PCR and Western blotting analysis. The proliferation of HCCLM3 cells was tested by MTT assay. Apoptosis ratio of HCCLM3 cells was detected by TUNEL method and flow cytometry (FCM). Expressions of PCNA and wide-type p53 protein were analyzed by immunocytochemistry (ICC) methods. Results: The sequence-specific shRNA (pshPPARγ) efficiently blocked the expression of PPARγ mRNA by 80.5%. At 48 h after transfection of pshPPARγ, proliferation of HCCLM3 cells was significantly suppressed by 71.5%. The positive rate of PCNA expression was (23.8 ± 7.2)% at 40 h transfection. The apoptotic rates were (24.2 ± 4.7)% as detected by TUNEL assay and (23.2 ± 4.2)% of cells as measured by FCM test, respectively. The detection results of the two methods were consistent. In pshPPARγ transfection group, cell cycle of HCCLM3 cells was arrested in G0/G1 phase and the proportion of cells in G2/M phase decreased. Moreover, expression of wide-type p53 protein increased significantly. Conclusion: Knockdown PPARγ expression with RNA interference technology can significantly suppress proliferation and induce apoptosis of HCCLM3 cells. It is related with up-regulation of wide-type p53 protein expression.

Objective: To observe the association of the expression of peroxisome proliferator-activated receptor γ (PPARγ) with microvessel density (MVD) and vascularization mimicry (VM) in hepatocellular carcinoma (HCC) and explore their clinical significance. Methods: Thirty-four cases of HCC confirmed by pathological examination between 2003 and 2006 in Nantong Tumor Hospital were recruited in this study. The expressions of PPARγ, MVD and VM were tested by CD34 immunohistochernical (IHC) staining and seriodic acid-schiff (PAS) reaction in 34 cases of HCC tissues and its paired non-cancerous tissues. Their biological behaviors were analyzed and associated with the clinicopathological features. Results: PPARγ was over-expressed (24/34) in HCC tissue. It was associated with tumor size (>5 cm, P =0.027), broken capsule (P =0.014),tendency to metastasis and recurrence (P =0.001). The MVD was higher (23/24) in HCC tissue than that in corresponding para-cancerous tissues (P < 0.05). High MVD was related with tumor size (>5 cm, P =0.010), broken capsule (P =0.001), tendency to metastasis and recurrence (P=0.004), with or without metastasis during follow- up period (P =0.008), and differentiation degree (P =0. 001). The positive expression of VM was detected in 7 out of 34 HCC tissues. The MVD was higher in VM-positive group (P < 0.05). The positive expressions of VM were associated with TNM stage (P =0.001), tendency to recurrence (P =0.029), and differentiation degree (P =0.004). There was a significantly positive correlation between the over-expression of PPARγ and high MVD (r = 0.519). Kaplan-Meier analysis indicated that positive expression of PPARγ and VM as well as high MVD could be used for predicting the survival rate of HCC patients (P =0.013, 0.036 and 0.022, log-rank test). Cox hazard model analysis showed that positive PPARγ expression and high MVD were the independent prognostic factors for HCC. Conclusion: The PPARγ correlates with the development of microcirculation in HCC. PPARγ is a novel prognostic indicator for evaluating the prognosis of HCC.

Vascular calcification (VC) is a chronic systemic vascular disease characterized by abnormal deposition of hydroxyapatite minerals in the vascular system and is closely associated with aging, diabetes, atherosclerosis, and chronic kidney disease. Perivascular adipose tissue (PVAT), a special type of adipose tissue that surrounds blood vessels, is thought to be a supportive component of the vascular structure and is capable of playing a role in homeostatic regulation during vasodilatation and contraction. Currently, there is growing evidence that perivascular adipose tissue acts as an endocrine and paracrine organ and interacts closely with cellular components of the vascular wall, which may be involved in the development of vascular calcification. This article reviews the role of perivascular adipose tissue in the pathophysiological process of vascular calcification and its potential as a target for therapeutic intervention, with the aim of providing new ideas for the prevention and treatment of vascular calcification.

To explore the clinical value of contrast-enhanced ultrasound (CEUS) in differentiating benign and malignant focal liver lesions (FLLs) with SonoVue, CEUS was used to examine 113 patients with focal liver lesions (FLLs) in our hospital during July 2005 to December 2006. All the patients underwent contrast-enhanced CT (CECT) or contrast-enhanced MRI (CEMRI). Except for patients with focal fatty sparings (n=18) and with hemangiomas (n=8), all the patients were confirmed by operation or ultrasonic-guided liver puncture biopsy. A sulfur hexafluoride gas-based contrast agent was used with a MI of 0.15 to 0.17. Forty-eight cases of malignant FLLs, including 30 hepatocellular carcinomas (HCCs), 2 cholangiocarcinomas and 16 metastatic tumors, were detected. Seventy-eight cases of benign FLLs, including 33 hemangiomas, 9 focal nodular hyperplasias (FNHs), 19 focal fatty sparings, 5 abscesses, 7 regenerative nodules and 2 inflammatory pseudo-tumor, were involved. The contrast pattern of benign and malignant FLLs was quite different. CEUS has higher specificity and sensitivity than conventional ultrasound in differentiating benign and malignant FLLs.
Carcinoma, Hepatocellular/*ultrasonography ; Contrast Media/*diagnostic use ; Diagnosis, Differential ; Hemangioma/ultrastructure ; Image Enhancement/*methods ; Liver Diseases/*ultrasonography ; Liver Neoplasms/*ultrastructure ; Magnetic Resonance Imaging ; Sensitivity and Specificity ; Young Adult

Objective To evaluate the relationship between transplantation of allogeneic peripheral blood stem cells (allo-PBSCs) and low incidence of leukemic relapse. Methods Thirty-seven adult patients with acute myeloid leukemia in first remission ( n =16) and chronic myeloid leukemia in chronic phase ( n =21) underwent allo-PBSCT from June 1997 to December 1999. A median number of CD34 + cells, CD3 + cells, CD4 + cells and CD8 + cells infused was 7.3 ?10 6/kg, 364?10 6/kg, 210?10 6/kg and 137?10 6/kg, respectively. Busulfan and cyclophosphamide were used as conditioning regimen (BU-CY2). All patients received cyclosporine and methotrexate for GVHD prophylaxis. Results Engraftment of neutrophil and platelet was achieved at a median of day +13(9～28) and day +12(7～19) respectively. Acute GVHD (aGVHD) occurred in 18 of 37 patients ( 48.6% ) with grade II～IV aGVHD ( 24.3% ). Chronic GVHD (cGVHD) developed in 22 (extensive 8, limited 14) out of 32 evaluable patients ( 68.8% ). 28 patients were still alive in complete remission at a median follow-up of 950 days (range 600～ 1 500 days). The main causes of death were aGVHD in two patients, CMV-IP in four patients, cGVHD in two patients, and relapse only in one patient. The probability of disease free survival at 3 years was 75.7% . Conclusion In our series, leukemia recurrence was seen only in one patient ( 2.7% ), suggesting that a high incidence of cGVHD may have an enhanced graft-versus-leukemia effect and hence reduce the incidence of relapse after transplantation.

Aim To study the anti-inflammatory activity of the Channa argus bile.Methods The bile was isolated and purified by extraction and silica gel column chromatography.Then the compounds were identified by hydrogen and carbon spectra.The spleen lymphocytes proliferation assay and Lipopolysaccharide(LPS) induced mouse macrophage RAW264.7 releasing Nitrogen Monoxide(NO) experiment were used to evaluate the anti-inflammatory activity.Results Compound(C1) of sodium taurocholate and compound(C2) of sodium taurochenodeoxycholate were isolated by activity tracing.The cell relative viabilities of the two compounds on Concanavalin A(Con A) induced spleen lymphocytes proliferation assay were 65.9%±11.7% and 60.5%±9.4%, which were significantly different from the result of model group (P<0.01), respectively.The NO production of LPS-induced RAW264.7 release of NO was (16.4±1.9) μmol·L-1 and (15.5±1.7) μmol·L-1, which were significantly different from the result of model group(P<0.01).Conclusion Sodium taurocholate and sodium taurochenodeoxycholate from Channa argus perform the anti-inflammatory activities but have no cytotoxic effect on spleen lymphocytes and macrophage.

Objective: To evaluate the analgesic effect of pregabalin combined with intrathecal sufentanil infusion for the treatment of breakthrough pain in patients with bone metastases. Methods: A total of 60 breakthrough pain patients with bone metastases were randomly divided to 3 groups: group A (pregabalin combined with intrathecal sufentanil infusion group,n=20), group B (placebo combined with intrathecal sufentanil infusion group,n=20) and group C (oral morphine sulfate controlled-release tablet group,n=20). The differences in visual analogue scale (VAS)between background pain and breakthrough pain, the seizure frequency of breakthrough pain, general satisfaction and side effects of the 3 groups were observed. Results: hTe seizure frequency and VAS of breakthrough pain in group A decreased signiifcantly after the treatment (P<0.05) and the general satisfaction was the best among the the 3 groups (P<0.05), with less nausea and vomiting, constipation, drowsiness and fewer other side effects. Conclusion: Pregabalin combined with intrathecal sufentanil infusion can effectively relieve breakthrough pain in patients with bone metastases.

Objective To study changes of serum IL-17 and IL-35 levels in patients with heart failure. Methods 60 patients with heart failure (observation group)were selected as research subjects.60 patients accord-ing to different severity were divided into acute period heart failure (34 cases)and stable stage heart failure (26 ca-ses);60 patients graded according to the NYHA standards were divided into 24 cases of heart failure with grade Ⅱ, 20 cases of grade Ⅲ ,16 cases of grade Ⅳ.According to the different primary diseases :expansion cardiomyopathy group (20 cases in group A),the coronary heart disease group (group B,24 cases),hypertensive heart disease group (group C,16 cases).During the same period,42 healthy elderly people in our hospital were selected as control group. The serum IL-17,IL-35 levels were tested,and the serum IL-17,IL-35 levels in patients with heart failure were ana-lyzed.Results Serum level of IL-17 in the observation group was higher than the control group,and the difference was significant [(15.61 ±4.02)pg/mL vs (9.49 ±3.96)pg/mL,t =9.018,P <0.01].Serum level of IL-35 in the observation group was significantly lower than that of the control group,and the difference was significant[(52.78 ± 4.29)pg/mL vs (61.49 ±4.81)pg/mL,t =11.963,P <0.01].The level of serum IL-17 in acute stage of patients with heart failure was higher than that of stable heart failure,and the difference was significant (t =6.278,P <0.01);IL-35 level in serum of patients with heart failure in acute phase was lower than that of stable heart failure,the difference was significant (t =9.529,P <0.01).With the increase in heart failure grade,serum IL-17 level showed a rising trend,and the differences among three groups had statistical differences (F =6.098,P <0.01);serum IL-35 level decreased,and the differences among three groups had statistical differences(F =8.978,P <0.01).The serum IL-17 level of A group was higher than that in B group and C group,there were significant differences (F =6.096, P <0.01),the serum IL-17 level between B group and C group had no statistical difference (t =0.172,P >0.05). The serum IL-35 level of A group was lower than that of B group and C group,there were significant differences (F =8.978,P <0.01),the serum IL-35 level between B group and C group had no statistical difference (t =0.208,P >0.05).Serum IL-17 and serum IL-35 level was negatively correlated (r =-0.429,P =0.009).Conclusion High expression of IL-17 in elderly patients with heart failure,while IL-35 decreased in elderly patients with heart failure, IL-17,IL-35 are closely related to the senile congestive heart failure and the severity of illness.Serum IL-17 is nega-tively correlated with the level of serum IL-35.

Objective To study the influence on allograft function of HLA antibody in patients who received pairs of allograft from the same donor.Methods In Beijing Friendship Hospital.HLA antibodies were tested from October,2008 to April 2009 in patients.Recently (October,2013-February,2014),renal functions(serum creatinine/urea nitrogen)were studied in 226 patients who received transplant from 113 donors.LATM10x5,One Lambdas used for Panel reactive antibody screen-ing.Results 41 pairs of renal for male,21 pairs of renal for female and 51 pairs of renal for both male and female.PRA posi-tive in 26 patients (only 4 pairs of renal for patients were positive),11 recipients (HLA II antibody positive in only 1 pair of renal for patients)and 36 recipients (only 5 patients produced antibody)in 226 patients,HLA antibody positive in 73 pa-tients,in which renal function lost or decreased in 64 patients.HLA antibody negative in 153 recipients,in which renal func-tion lost or decreased in 4 patients.There was significant difference between the two group (χ2=160.70,P<0.001).Con-clusion HLA antibody is a important factor influence renal function and long term survival.

Objective:To optimize the determination technology for the total amino acids in Runing pills. Methods:The process was optimized by L9 (34 ) orthogonal design using the hydrolysis temperature, hydrolysis time and hydrochloric acid concentration as the evaluation factors,and the content and yield of total amino acids as the evaluation indices. Results: The optimal determination condi-tions were as follows:the hydrolysis temperature was 110℃, the hydrolysis time was 22 h and the hydrochloric acid concentration was 6 mol·L-1 . Conclusion:The optimal determination technology can be applied in the total amino acids from Runing pills,which pro-vides the basis for the further research of Runing pills.