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.

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.

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

OBJECTIVETo investigate the clinical characteristics of Segond fracture combined with injuries, and to explore the operative method and opportunity.

METHODSFrom June 2010 to December 2014, 10 patients with Segond fracture were treated. There were 10 males, ranging in age from 26 to 69 years old, with a mean of 42 years old. Under arthroscopy examination within 7 to 10 days after injury, 6 patients had compete rupture of anterior cruciate ligament and meniscus injury; 2 patients had tibial avulsion fractures of the ACL combined with meniscus injury. At 4 to 8 weeks after injury, 2 patients were performed with arthroscopy examination; 1 patient had complete rupture of anterior and posterior cruciate ligament combined with meniscus injury; and the other patient had complete rupture of anterior cruciate ligament alone. All the patients with cruciate ligament rupture were treated with reconstruction of cruciate ligament; 9 patients had meniscus injuries were treated with menisus plasty or suture; and 2 patients had tibial avulsion fractures of the ACL were treated with non-absorbable Ethicon suture fixation under arthroscopy. After operation, the Lysholm scoring system was used to evaluate clinical effects.

RESULTSEight patients were operated within 10 days after injury, the knee joints swelling was obvious at the 3rd day after operation, and blood oozing was found by joint puncture. These 8 patients did out-of-bed activity with assistance at 1 week postoperatively. Other 2 patients performed operation in 4 to 8 weeks after injury without knee joint obvious swelling at the 3rd day after operation, and no blood oozing was found by joint puncture. These 2 patients did out-of-bed activity with assistance after 3 days postoperatively. All the patients were followed up, and the duration ranged from 12 to 24 months, with an average of 18 months. Postoperative Lysholm score of all patients was higher than preoperative scor, and the results were satisfactory.

CONCLUSIONFor Segond fractures, firstly, it is important to gain early diagnosis for treatment. Secondly, it is often associated with the rupture of anterior cruciate ligament and meniscus injuries, and the best time of cruciate ligament rebuilding and meniscus repairing maybe between 4 to 8 weeks after injury. Thirdly, personalized treatment should be chosen according to different complications.

Adult ; Aged ; Anterior Cruciate Ligament Injuries ; Arthroscopy ; Humans ; Male ; Middle Aged ; Tibial Fractures ; complications ; diagnosis ; surgery ; Tibial Meniscus Injuries

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.

AIM: To explore the effect of berbamine(BER) on apoptosis in K562 cells and its possible molecular mechanisms. METHODS: The apoptosis rate was measured by flow cytometry while electron microscopy and DNA electrophoresis were used to evaluate the characteristic changes of apoptosis, RT-PCR and Western blot were used to examine the expression levels of apoptosis related gene bcr/abl and BCR/ABL protein. RESULTS: By FCM, the apoptosis rate of K562 cells treated with 8.0 mg/L BER for 24 h and 72 h increased from (29.20?3.82)% to (61.77?4.35)% (P

Objective Off-pump coronary artery bypass grafting (OPCAB) is used more widely in recent years in China. However, there is an argument on benefits and risks of off-pump surgery. Many studies shown that OPCAB had more benefits in short-term outcomes than conventional coronary artery bypass grafting(CCABG). But evidences from other studies suggested that OPCAB resulted in less long-term graft patency as compared with on-pump surgery. This study examined the longterm graft patency of OPCAB and CCABG performed by one surgeon. Methods 50 patients who had received surgical revascularization by a surgeon for more than 5 years were reviewed, 25 patients received conventional coronary artery bypass grafting ( group 1 ) and 25 patients received OPCAB ( group 2). All patients had angiograms for compareing the graft patency between the two groups. Results Among 25 patients in group 1,21 were male and 4 were female. The mean age of patients at surgery was (55.4 ±8.9) years. 15 cases had unstable angina, 16 patients had old myocardial infarction and 6 cases had diabetes.The ejection fraction (EF) was 0.58 ±0.14. The mean number of bypasses per patient was 3.32 ±0.63. Mean duration of operation was (3.58 ± 0. 82) hours. Mean follow-up duration was ( 86.52 ± 12.48) months. 83 grafts were evaluated for patency ( open vs. closed) and were graded by Fitzgibbon as grade A ( excellent graft), B ( impaired graft, with a stenosis of ≥50%, or a diameter less than 50% of the grafted artery), or O ( completely occluded). The graft patency was 73.49%, 61grafts were graded as Fitzgibbon A, 6 grafts as Fitzgibbon B and 16 grafts as Fitzgibbon 0. 25 patients were in group 2, 21males and 4 females. The mean age of patients at procedure was (58.2 ± 9.09) years, 11 patients had unstable angina, 13 patients had old myocardial infarction and 6 cases had diabetes. The ejection fraction (EF) was 0.59 ± 0. 14. Conclusion No 2011.03.013 difference in long-term graft patency was identified between on-pump and off-pump coronary artery bypass grafting. Off-pump oronary artery bypass grafting preformed by an experienced surgeon may gain similar long-term graft patency to that of conventional bypass.

Objective To investigate the clinical skill and experience of laparoscopic herniorrhaphy in the treatment of abdominal hernia. Methods Clinical data of 30 cases of abdominal hernia undergoing laparoscopic her-niorrhaphy in Shengjing Hospital from May 2004 to December 2008 were analyzed retrospectively. There were 23 in-guinal hernia,4 incision hernia and 3 hiatal hernia. TEP in 20 cases,TAPP in 3 cases,lPOM in 4 cases and Nissen hiatal hernia repair in 3 cases were performed. Results The mean operating duration was 72 min for TEP, 127 for TAPP,150 min for IPOM,and ]60 min for hiatal hernia repair and the mean time for hospitalization was 4.6,5.8, 7.5, and 5.3 days. No conversion to open surgery happened among these cases. No recurrence was found. Conclu-sions Laparoscopic herniorrhaphy appears to be an effective and convenient technique for the treatment of abdomi-nal hernia and has many advantages of minimally invasive surgery.

Objective To discuss the perioperative nursing for patients with Laparoscopic resection of gastrointestinal stromal tumors. Methods A retrospective study was carried out in 18 patients with gastrointestinal stromal tumors on the effect of perioperative nursing and surgery. Results The gastrointestinal stromal tumors were resected under Laparoscopy with average operation time of 70 minutes. Blood loss volume during the operation was 20 to 50 ml. All patients recovered well with hospitalization days of 4 to 9 days. No relapse or metastasis was seen after 6 to 22 months of follow-up visit. Conclusions Laparoscopic resection of gastrointestinal stromal tumors was safe and effective. Perioperative nursing had significant effect on the recovery of patients.