Objective To investigate the effects of down-regulation of Netrin-1 expression on cell proliferation,apoptosis and migration of a cutaneous squamous cell carcinoma(CSCC)cell line SCL-1.Methods Unspecific(control)small interference RNA(siRNA)and Netrin-1-targeting siRNA were designed and constructed.SCL-1 cells were divided into 3 groups to remain untreated(control group 1),be transfected with control siRNA(control group 2)and Netrin-l-targeting siRNA(experiment group)by using Lipofectamine 2000 respectively.After 48 hours of additional culture,flow cytometry was performed to detect the apoptosis of cells,Western blot to determine the protein expression of caspase-3 and-9,Netrin-1 and matrix metalloproteinases (MMP)in these cells,and Boyden chamber was utilized to evaluate the migration of cells.Enzyme linked immunosorbent assay(ELISA)was performed with cholecystokinin-8(CCK-8)kit to estimate the proliferation of cells at 24,48,72 and 96 hours after the transfection.Results The expression of Netrin-1 protein was significantly downregulated in SCL-1 cells in the experiment group compared with the control group 1 and 2 (0.207 ± 0.044 vs.0.741 ± 0.111 and 0.716 ± 0.109,F =31.43,P ＜ 0.05).The Netrin-1-targeting siRNA significantly downregulated the proliferation of SCL-1 cells,but induced the apoptosis in SCL-1 cells with significant differences in the percentage of early apoptotic cells and late apoptotic cells between the control group 1,2 and experiment group(11.74 ± 0.33 vs.11.55 ± 1.22 vs.24.74 ± 2.43,F=68.63,P＜ 0.01;3.82 ± 0.32 vs.4.69 ± 0.68 vs.5.33 ± 0.18,F =8.58,P ＜ 0.05).The experiment group showed a significant increase in the expression of caspase-3 and-9(1.458 ± 0.111 vs.0.398 ± 0.057 and 0.412 ± 0.049,F =183.99,P ＜ 0.01;0.967 ± 0.099 vs.0.234 ± 0.036 and 0.205 ± 0.033,F =138.11,P ＜ 0.01),but a statistical decrease in the expression of MMP-2(0.235 ± 0.038 vs.0.862 ± 0.040 and 0.892 ± 0.035,F =291.07,P ＜ 0.01)compared with the control group 1 and 2.Conclusions The down-regulation of Netrin-1 expression can suppress the proliferation and migration of SCL-1 cells,but promote the apoptosis in SCL-1 cells.

Objective:Toprepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods: The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP-1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR, and the proliferation of HepG2 cells was detected by MTT assay. Results: The microsphere encapsulating recombinant TIMP-1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×10~8/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP-1 PELA microsphere efficiently induced TIMP-1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion: PELA microsphere encapsulating recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.

Objective To investigate plasma levels of total carnitine (TC) and free camitine (FC) in patients undergoing hemodialysis and peritoneal dialysis.Methods 200 cases of normal group came from physical examination in this hospital,all testing cases were the in-hospital patients in the department of nephropathy.TC and FC were determined by use of an enzymatic cycle assay on Hitachi 7170 automatic biochemical analyzer.Results In 200 cases of normal group,TC level was (56.52?9.61) ?mol/L,and FC was (46.60?8.23) ?mol/L.In 37 hemodialysis patients,TC and FC levels were (41.47?13.22) ?mol/L and (24.58?8.91)?mol/L before dialysis,a statistic difference was observed against the control group (P0.05).Conclusions Carnitine deficiency was seen in most patients undergoing hemodialysis and peritoneal dialysis.Furthermore,the deficiency status got worse along with the dialysis course in hemodialysis patients.Carnitine infusion can effectively improve the status of these patients.

OBJECTIVETo explore the correlation of the gene polymorphisms of Toll-like receptor 2 ( TLR2) and TLR4 with the susceptibility and recurrence of condyloma acuminatum (CA).

METHODSUsing Snapshot, we detected the gene polymorphisms of TLR2 597(T/C), 1350(T/C), 15607(A/G), and 2258(G/A) and TLR4 896(A/G) and 1196(C/T) in the peripheral blood of 140 CA patients and 105 HPV-negative controls. We made comparisons between the CA patients and controls as well as between the cases of recurrent CA and those of non-recurrence at 6 months after treatment.

RESULTSThere were 72, 48, and 20 cases of genotype TT, TC, and CC of TLR2 597 (T/C), respectively, in the CA patients, as compared with 71, 31, and 3 cases in the controls. The gene frequency of mutant C was 31. 43% in the patients, significantly higher than 17.62% in the controls (χ2 = 12.04, P < 0.01), and it was 38.68% in the recurrent cases, remarkably higher than 27.01% in the non-recurrent cases (χ2 = 4.16, P < 0.05). There were 74, 49, and 17 cases of genotype TT, TC, and CC of TLR2 1350( T/C), respectively, in the CA patients, as compared with 73, 29, and 3 cases in the controls. The gene frequency of mutant C was 29. 64% in the patients, significantly higher than 16. 67% in the controls (χ2 =11.05, P < 0.01), and it was 36.79% in the recurrent cases, markedly higher than 25. 29% in the non-recurrent cases (χ2 = 4.18, P < 0.05). There were 44, 66, and 30 cases of genotype AA, AG, and GG of TLR2 15607(A/G), respectively, in the CA patients, as compared with 26, 58, and 21 cases in the controls. There was no significant difference in the gene frequencies of mutant G between the two groups (χ2 = 0.33, P > 0.05). No mutant genes of TLR2 2508 (G/A) or TLR4 896(A/G) and 1196(C/ T) were detected in either the CA patients or the controls. Linkage disequilibrium analysis showed a tight linkage between TLR2 597 (T/C) and 1350(T/C) (D' = 1, r2 = 0.93).

CONCLUSIONTLR2 597(T/C) is tightly linked to 1350(T/C), which is correlated with both the susceptibility and the recurrence of condyloma acuminatum.

Aged ; Case-Control Studies ; Condylomata Acuminata ; genetics ; Gene Frequency ; Genetic Linkage ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Recurrence ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics

ObjectiveTo evaluate the various methods of choledochoplasty in the repair of major bile duct defects in Mirizzi syndrome.MethodsThis is a retrospective study on 3 patient with Mirizzi syndrome with a large bile duct defect.These defects were repaired by using a vascular gastric pedicle patch in our department from July 2008 to November 2011.The authors searched the domestic medical literature on surgical repair for Mirizzi syndrome in the past ten years.The patients were treated by various surgical methods,and they were analyzed according to the Csendes Classification.ResultsThere were no surgical complications in our three patients.There was one patient with a Csendes type Ⅲ,while the remaining 2 patients were with Csendes type Ⅳ.At a median follow- up of 2.5 years,no patient developed signs of chronic cholangitis.In the medical literature,there were 93 patients who were with Csendes type Ⅰ ; and 58 patients were treated by cholecystectomy only,while 35 patients were treated by partial cholecystectomy plus mucosal ablation.Of the 40 patients with type Ⅱ,29 patients were treated by direct fistula repair,9 patients by pedicle gallbladder flap and 2 patients by pedicle round ligament.Of the 20 patients with type Ⅲ,9 patients were treated by pedicle gallbladder flap.1 patient by pedicle round ligament,3 patients by pedicle gastric flap and 7 patients by Rouxen- Y hepaticojejunostomy.For the 5 patients with type Ⅳ,they were treated by Roux-en- Y hepaticojejunostomy.Of these 159 patients,postoperative complications included biliary fistula (n=1 ),upper gastrointestinal bleeding (n=1),and biliary stricture (n=1).All the remaining patients were cured.ConclusionIn patients with Mirizzi Syndrome,the choice of treatment depends on the size of the fistula.For patient with a major tissue defect in the common hepatic duct,a pedicle vascular gastric flap is a good treatment.

In order to find the anti-virus constituents of Alternanthera philoxeroides (Mart.) Griseb, the investigation was carried out. The paper reported the five triterpenoid saponins isolated from n-BuOH fraction: 3-O-beta-D-glucopyranosyl (1-->3)-O-[beta-D-glucopyranosyl-oleanolic acid]-28-O-beta-D-glucuronopyranoside (1), oleanolic acid-3-O-beta-D-glucuronopyranoside (calenduloside E, 2), oleanolic acid-3-O-beta-D-glucopyranosyl-28-Obeta-D-glucopyranosyl ester (chikusetsusaponin-IVa, 3), 3-O-(6'-O-butyl-beta-D-glucuronopyranosyl)-oleanolic acid-28-O-beta-D-glucopyranosyl ester (4) and hederagenin-3-O-beta-D-glucuronopyranoside (HN-sapoins K, 5). 1 is a new compound, saponins 4 and 5 were isolated from the plant for the first time.

Objective To discuss the feasibility and effects of cemented artificial bipolar arthroplasty for senile patients with femoral intertrochanteric instable fractures. Methods From January 2004 to June 2008,46 aged cases with femoral intertrochanteric instable fractures were cured with cemented artificial bipolar arthroplasty. According to improving-classfication of Evans,there were 19 cases with type Ⅲfractures and 27 cases with type Ⅳ fractures. Results All cases were successfully operated. The surgery time was 50-90 (68.00 ±20.43) minutes. Bleeding volume was 160-600 (210.00 ±40.26) ml. The time in hospital was 16-26 (19.30 ± 4.56) days. All cases were retrospectively reviewed for 6-39 (17.10 ± 4.33)months. Three cases died of other diseases, 43 cases were graded by the system of Harris. The Harris scale was 70-94 (86.70 ±5.16) scores,and the choiceness rate was 90.7%(39/43). No dislocation,loosening,subsidence and late infection occurred in all the implants. Conclusions Cemented artificial bipolar arthroplasty has good advantages of reduced laying up period and few complications. The short-term outcome is satisfactory.

Objective:To investigate the inhibitory effects of melanoma differentiation associated gene-7(mda-7/IL- 24)on lymphoma cell line Namalwa in vitro and in vivo.Methods:Using RT-PCR,the expression of mda-7/IL-24 was examined in 10 malignant hematopoietic cell lines,including Namalwa,Raji,K562,NB4,U937,Ramous,CEM,KG1a, HL60,J6-1,etc.The coding region of mda-7/IL-24 was cloned from LPS-treated human peripheral blood mononuclear cells(PBMC)by RT-PCR,and the eukaryotic expression vector pTarget-IL-24 was constructed.The recombinant vector, after sequenced,was transfected into Namalwa cell line via lipofectamine reagent.The stable expression transfectants were selected by G418.The expression of mda-7/IL-24 mRNA and protein was verified by RT-PCR and Western blotting.MTT assay,colony forming assay,apoptosis detection,and tumorigenesis in nude mice were used to assess the effects of mda- 7/IL-24 on tumor proliferation,growth characteristics,colony forming,apoptosis,and tumorigenesis.Results:Expression of mda-7/IL-24 mRNA was not found in any of the 10 malignant hematopoietic cell lines and the expression of mda-7/IL- 24 mRNA and protein was found in Namalwa cells transfected with recombinant plasmid pTarget-IL-24.Significant de- crease in tumor cell viability was observed in Namalwa cells stably transfected with mda-7/IL-24,compared with control cells transfected with empty plasmid pTarget(P

BACKGROUND: With the increasing number of patients undergoing knee arthroplasty in China, the complications caused by the mismatch of prosthesis with anatomical parameters arouse extensive attentions.OBJECTIVE: To measure the anatomical parameters of the knee joints in healthy Zhuang ethnic group from Guangxi Zhuang Autonomous Region, China, using three-dimensional (3D) reconstruction technology, thus providing reference for the design and placement of the hip prosthesis.METHODS: Sixty-eight individuals, including 30 females and 38 males, were selected from 217 healthy people undergoing CT examination. All data were imported into Mimics 15.0 software used for 3D reconstruction, and then the knee parameters were then measured and analyzed.RESULTS AND CONCLUSION: There was no significant difference in the parameters of the bilateral femur and tibia (P > 0.05). The femoral parameters had significant differences between genders (P < 0.05). The width of tibial platform, sagittal length of lateral tibial platform, distance between medial tibial plateau and fibular head, and posterior slope of tibial plateau showed no significant differences between genders (P > 0.05), while there were significant differences in the width of medial/lateral tibial platform, and sagittal length of medial tibial platform between genders (P < 0.05). The sagittal length of medial femoral condyle was negatively correlated with age, and all femoral parameters were positively correlated with height; while only the width of medial and lateral tibial platform were positively correlated with height. To conclude, 3D reconstruction technology is available for research on the morphology of the knee joint in the Zhuang ethnic group from Guangxi Zhuang Autonomous Region, which provides references for prosthesis design and research appropriate for the Zhuang ethnic group.

Human herpes simplex virus 1 (HSV-1) causes facial,ocular,and encephalitic disease and is associated with latent infection and cancer.Here,we developed a means of studying the pathogenesis of HSV-1 infection at the protein level by using the SELDI Protein Chip to detect changes of protein expression in Vero cells cultured in vitro.After infection with HSV-1 and culture for 12,24 or 48 h,cells were harvested and lysed.IMAC3 arrays were applied to SELDI-TOF-MS to detect proteomic differences before and after infection.The chip detected a series of differentially expressed protein peaks.Interestingly,both peaks at 16 912 Da and 17 581 Da corresponded precisely with the molecular mass of ISG 15,which may participate in antiviral activity during the process of infection.Thus,the results we obtained can serve as a basis to study the pathogenesis of HSV-1 and the interaction between the virus and its host.In addition,they can help in the discovery of new therapeutic targets for treatment of HSV-1 infection.