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.

Objective To investigate the influences of HLA mismatching on renal function in the kidney transplant patients re-ceiving pairs of allograft from the same donor.Methods 171 pairs of renal transplant patients receiving the kidneys from the same donors were investigated.They were admitted in our hospital before 2008.Their human leukocyte antigens (HLA)were typed with the commercial polymerase chain reaction (PCR)-sequence-specific primers (SSP)HLA typing kit (One Lambda,Inc.,USA;and GTI Diagnostics,USA).The serum creatinine (SCr)and blood urea nitrogen(BUN)were measured in the clinical laboratory of our hospital.Results Among 171 pairs of renal transplant patients,there were 162 recipients with HLA mismatch≤4,in which the re-nal function was remained stable in 107 recipients and lost or decreased in 55 patients.There were 180 recipients with HLA mis-match >4,in which the renal function was stayed normal in 84 recipients and lost or decreased in 96 patients.The difference in in-fluencing the renal function between the HLA mismatch≤4 and HLA mismatch>4 had statistical significance (χ2 =12.22,P <0.05).Conclusion Excellent HLA typing match has important significance for renal 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.

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 observe the effect of hot spring medicated bath regimen which is guided by the theory of constitution distinction of TCM.Methods According to the standard of Constitution Classification and Decision of TCM which is published by China Association of Chinese Medicine to assess the standard-compliant volunteers,then make the volunteers have physical-chemical indexes checked and hot spring medicated bath taken.At last,re-assess the constitution of the individual and re-check the indexes.Results After the drug bath,the levels of total cholesterol (TCH),low-density lipoprotein (LDL-C),thyroid stimulating factor(TSH),and IL-1 of the patients who were both yang deficiency and phlegm-dampness decreased significantly (t values were 2.093,2.045,2.092,and 2.13,respectively; P＜0.05); the levels of high-density lipoprotein (HDL-C),cortisol,and free thyroxine T4 (fT4) increased significantly (t values were 3.731,2.571,and 3.027,respectively; P＜0.05) ; the pulse rate raised obviously (t=8.323,P＜0.01) ; both of the systolic (t=4.405,P＜0.01) and diastolic (t=2.782,P＜0.01)blood pressure dropped.After the drug bath,the levels of fT4 and IL-1 of the patients who were yang deficiency significantly increased (t values were 3.709 and 3.339,respectively; P＜0.01); the waist circumference reduced (t=2.831,P＜0.01),the pulse rate rised (t=9.605,P＜0.01) ; both of the systolic (t=5.281,P＜0.01)and diastolic (t=6.573,P＜0.01)blood pressure dropped.After the drug bath,the levels of pulse rate and diastolic blood pressure of the patients who are phlegm-dampness changed significantly:the pulse rate raised (t=4.702,P＜0.01),the diastolic blood pressure dropped(t=6.096,P＜ 0.01).As the effect analysis showed,the Ridit data of the criterion group which consists of summation was 0.5.Compared with the criterion group,the total effective rate of the yang deficiency group was 74.1％,which was statistically significant (R=0.55,P＜0.05).The total effective rate of the phiegm-dampness group was 81.8％,which had no statistical significance (R=0.48,P＞0.05).The total effective rate of the both yang deficiency and phlegm-dampness group was 77.4％,which was statistically significant (R=0.46,P＜0.05).Conclusion The Wuyi hot spring medicated bath regiman is helpful to the constitution type of yang deficient and phlegm-dampness,which can help warming yang and benefiting qi,living blood links contact us nourishing the skin,reducing blood fat,lowering blood pressure,improving the immune function and so on.

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.