ObjectiveTo study the effect of TGF-? 1 signal transduction blocked by antisense-Smad 4 gene on progression of experimental liver fibrosis of Kunming mice.MethodsWe transfered retrovirus-mediated antisense Smad 4cDNA into the CCl4/ethanol induced cirrhotic liver of Kunming mouse model to investigate the antisense-Smad 4 gene integration by Southern blot. The expression of Smad 4 in cirrhotic liver was observed by Northern blot, RT-PCR, and Western blot. The fibrotic degree of the livers among the three groups were compared.ResultsThe antisense-Smad 4 gene was confirmed to integrate into the liver of the retrovirus-treated mouse .In the non-treated cirrhotic liver, the expression of Smad 4 mRNA was significantly increased than normal liver, the antisense Smad 4 gene could decrease the expression of Smad 4. Compared with the non-treated liver, the fibrotic septa of the liver in the retrovirus-treated mouse were fewer,narrow, and incomplete,and their fibrotic degree of the treated-liver was reduced.ConclusionThese results indicate that antisense-Smad 4 gene can reduce the expression of Smad 4,inhibit the progression of liver fibrosis.

Objective To evaluate the clinical effect of early total care (ETC) for treatment of unstable pelvic fractures.Methods This retrospective case series study included eighteen patients with unstable pelvic fractures treated surgically from May 2013 to May 2015.There were 11 male and 7 female patients,aged (45.1 ± 13.1)years.Eleven patients were injured due to road traffic accidents and 7 due to fall from height.Tile B type pelvic fractures were seen in 8 patients and Tile C type in 10 patients.All patients were treated with ETC method and received open reduction and internal fixation within 24 to 48 hours after the primary treatment.Data were collected,such as operation time,intraoperative blood loss,hospital length of stay and perioperative complications.Majeed score and EuroQol 5 dimension (EQ-5D) were used for evaluation of clinical outcome and quality of life respectively.Excellent and good rate of operation was assessed using the Matta clinical evaluation standard.Results Three patients died and fifteen patients survived.Operation time was (120.1 ± 22.2)minutes,amount of intraoperative blood loss was (355.9 ± 56.4) ml,and hospitalization was (8.5 ± 1.6) days.Incidence of perioperative complications was 56％,including 3 patients with acute respiratory distress syndrome,1 pneumonia,2 acute lung injury,1 incisional wound infection,2 multiple organ failure and 1 diffuse intravascular coagulation.All patients were followed up for (15.1 ±2.4)months (range,12-21 months).Majeed score was decreased from preoperative (93.7 ± 6.1) points to final follow-up of (74.1 ± 9.2) points,and the EQ-5D index was decreased from preoperative 0.96 ± 0.04 to final follow-up of 0.74 ± 0.19 (both P ＜ 0.05).Excellent and good rate of operation was 80％.Conclusions ECT provides satisfactory short-term outcome for treatment of unstable pelvic fractures,but the incidence of complications is high.Doctors should choose reasonable treatment plan according to the surgical indications.

Objective To investigate the construction and implementation of the concept of precise sequential therapy for primary liver cancer.Methods The clinical data of 207 patients with resectable liver cancer who were admitted to the Air Force General Hospital from May 2005 to June 2010 were retrospectively analyzed.Of all the patients,81 received conventional therapy (conventional therapy group),and the other 126 patients received precise sequential therapy (precise sequential therapy group).The conditions of the patients in perioperative phase,during postoperative sequential treatment and the period of follow-up between the 2 groups were compared.All data were analyzed by using the covariance analysis,analysis of variance,chi-square test or rank sum test.Results The detection rates of lesions with a diameter less than 1 cm,operation time,liver resection volume,intraoperative blood loss,rate of perioperative blood transfusion,duration of postoperative hospital stay,incidences of postoperative complications were 81％ (22/27),( 186 ± 36) minutes,(75 ± 29) ml,( 189 ± 60) ml,24％(30/126),(21 ± 12)days and 13％ (17/126) in the precise sequential therapy group,and 18％ (2/11),(222 ± 30)minutes,(133 ±88)ml,(327 ±46)ml,51％ (41/81),(26 ± 17)days and 20％ (16/81) in the conventional therapy group,respectively,with significant differences between the 2 groups (F =10.876,7.390,46.996,31.025,14.556,6.315,4.017,P ＜ 0.05).No significant difference was observed on the levels of alanine transaminase,albumin and the Child-Pugh score before and after the intervention in the precise sequential therapy group,but significant differences were observed in the conventional therapy group.The 1-,2-,3-year tumor recurrence rates and the 1-,2-,3-year survival rates were 17％ (21/126),22％ (17/76),26％ (8/31) and 87％ (110/126),87％ (66/76),84％ (26/31) in the precise sequential therapy group,and 31％ (25/81),38％ (27/71),48％(31/65 ) and 77％ (62/81),75％ (53/71 ) and 60％ (39/65) in the conventional therapy group,respectively.There were significant differences in the prognosis of the patients in the 2 groups ( x2 =4.958,4.292,4.168,4.062,3.640,5.470,P ＜ 0.05 ).Conclusion Through accurate assessment of the patients' condition before surgery,precise hepatectomy and precise postoperative intervention,the goal of effective control of tumor recurrence,maximum protection of the liver function and improvement of the survival rate can be achieved.

AIM: To construct eukaryotic expression vector of small interfering RNA(siRNA) specific to bcl-2 and investigate the effect of recombinant plasmid on suppressing bladder cancer cell growth.METHODS: siRNA of bcl-2 gene was designed according to the principle of RNAi-based medicine, and was converted into cDNA coding expression of small hairpin RNAs(shRNA) of siRNA. The cDNA was synthesized and inserted into plasmid pGenesil-1. The recombinant eukaryotic expression vectors of pGenesil-1545 and pGenesil-1555 were controlled by the U6 promoter of RNA polymerase Ⅲ, identified by the restriction map and the sequence analysis, and transfected into T24 cells. After T24 cells were transfected for 72 h, expression of bcl-2 mRNA was assayed by RT-PCR; and MTT was used to observe the proliferation of T24 cells.RESULTS: The recombinant plasmids of pGenesil-1545 and pGenesil-1555 were identified by the restriction map and the sequence analysis. The sequences completely coincided with the designs. The expression of the bcl-2 mRNA in T24 cells transfected with recombinant plasmid decreased nearly 80%, and the growth of T24 cells was suppressed significantly.CONCLUSION: The siRNA eukaryotic expression vector against bcl-2 gene is successfully constructed. It effectively downregulates the expression of bcl-2 in T24 cells and suppresses the cell growth.

Objective Combination of nanoparticle with p53 and Rb gene therapy by gene targeting was applied to investigate its curative effect and safety evaluation on colorectal rabbit hepatic VX2 metastasis for tumor eradication and survival enhancement. Methods Recombinant expressing plasmids harboring wild type p53 and Rb were cotransferred or transferred separately to the rabbit hepatic VX2 metastasis by the emulsion of PLL-nHAP nanoplex and lipodiol through the hepatic artery in a tumor target manner. Subsequent co-expressions of p53 and Rb protein within the treated tumors were detected by Western blot and in situ analysis of confocal laser scanning microscope. The therapeutic effect was evaluated by the tumor growth velocity and the survival time of animals. Eventually, investigations of liver function were applied to evaluate the safety of the process. Results With safe procedure for the rabbits liver function, both p53 and Rb local nano-therapy showed favorable anti-tumor effects and increased animal survival time. p53+Rb local nano-therapy could significantly inhibit hepatic VX2 metastasis and enhance the animal survival time compared with p53 local nano-therapy or Rb local nano-therapy. Local nano-therapy showed no significant influence to animal liver function. Conclusions Rb can work synergistically with p53 in the combined therapy mediated by PLL-nHAP nanoplex to augment the anti-tumor effect. The local nano-therapy with p53 and Rb is likely to be an effective and safe anti-tumor therapy for hepatic colorectal metastasis.

Objective: To investigate the cinlical features and therapeutic methods of unex-pected gallbladder carcinoma. Methods: A retrospective analysis was performed in 19 cases of unexpected gallbladder carcinoma treated in our hospital from April 2002 to April 2008. Results: Ten cases in early stage were perfomed gallbaldder resection and local lymph node removal, and the average survival time was 18.7 months; 4 cases were perfomed extend radical resection, and the average survival time was 12.3 months; 5 cases were performed palliative resection, and the survival time was less than 6 months. Conclusions: Frozen section examination during opera-tion is key to diagnose unexpected gallbladder carcinoma. Radical resection is the first option once the diagnosis is confirmed, and the method can be decided according to Nevin classification and B ultrosound scaning during opreation.

AIM: To identify the non-steroid transcription factors upregulating the expression of L-plastin in hormone-independent prostate cancer, and partly elucidate the mechanism of hormone-refractory prostate cancer. METHODS: TF SEARCH software was used to analysis the possible binding sites of transcription factors in the 3’ end of L-plastin promoter that had been identified as important part of regulation response elements. Gel shift assay and supershift assay were used to confirm the transcription factors binding the speculated response elements. PCR site-mutagenesis technique was performed to delete the binding site of transcription factor and luciferase activity assay was carried out after deletion of the binding site. RESULTS: SP-1 respond element GGTGGGGCGGGGA located at -54- -41 of L-plastin promoter was identified with the TF SEARCH software. Gel shift assay and supershift assay confirmed that SP-1 was the transcription factor binding to GGTGGGGCGGGGA. Mutant deleted the SP-1 binding-site had low-luciferase activity than that of the naive. CONCLUSION: SP-1 plays an important role in the up-regulation of L-plastin expression in hormone-independent prostate cancer.

Objective To verify the efficiency of a new laparoscopic urethrovesical anastomosis training model by comparing it with the chicken skin model. Methods Chicken posterior trunks and porcine colons were used to construct the training model. The posterior trunk of a chicken was used to simulate a human pelvis, and a 3-mm cloacal stump was used to simulate a human urethral stump. A 15-cm segment of porcine colon with a 1-cm orifice was used to simulate a human bladder or neobladder. An imitation urethrovesical anastomosis was performed with laparoscopic instruments in a laparoscopic training box. Forty trainees with no laparoscopic experience were randomized into 2 groups.The trainees in group A (n=20) practiced using this new model for 8 h, while those in group B (n=20) practiced using the chicken skin model for 8 h. The trainees' skills were assessed using the porcine model before and after training. Results Compared with the chicken skin model, this new training model more accurately resembled the structure and characteristic of human pelvis, urethral stump, and bladder (neobladder). After the training sessions, both groups improved in anastomosis time [GroupA: (64±11)min vs. (123±20)min, P＜0.05; Group B: (77±12)min vs. (121±17)min, P＜0.05] and quality (Group A: 8.8± 1.0 vs. 3.8 ± 1.2, P＜0. 05 ; Group B: 7.7 ± 0.9 vs.3. 7± 1.1, P＜0. 05). Compared with trainees in group B, trainees in group A required less time and achieved a higher quality score (P＜0.05). Conclusions This new training model can help urologic surgeons to reduce learning curve of this technique and improve their suturing skills. It is an effective,convenient training model for laparoscopic urethrovesical anastomosis.

Objective To compare superiorities of liver malignant tumors underwent regular and irregular hepatectomies through approaching perioperative factors.Methods 1019 consecutive hepatectomies of liver malignant tumor from 1986 to 2009 at Air Force General Hospital and General Hospital of Chinese PLA were investigated retrospectively according to their medical documentation.Results Multivariate analysis showed that liver malignant tumors on which regular hepatectomy and irregular hepatectomy were performed,there was no significant difference in the blood loss,complications,mortality related to operation,hospital stay,and so on.But the operating time of regular hepatectomy was obvious more than that of irregular hepatectomy (P＜ 0.001,OR=1.004).Conclusions Although for liver malignant tumor,regular hepatectomy seems to be superior to irregular hepatectomy based on oncological theory,in clinical practice,there were no significant difference between the perioperative risk of regular hepatectomy and that of irregular hepatectomy.

Objective: To explore the parameters used for the clinical evaluation of post-trau- matic hepatic function crashing down to the ground. Methods: By establishing the animal model of liver impact injury in simulation of parachuter,blood were sampled before injuries and 15 min, 30 min,45 min,1 h, 3 h, 6 h ,9 h after injuries. Cytokines including TNF?、IL-2 and liver function parameters including ALT,AST,LD-L,GGT,ALP were measured and analyzed statistically. Results: TNF? increased after 15 min- utes, and IL-2 increased after 30 minutes(P0.05). These two pa- rameters were significantly increased all the time after injuries(P