OBJECTIVETo investigate the apoptosis regulation on hepatoma cells by HBx between genotype B and C.

METHODSGenotype B and C HBx gene fragments were amplified and inserted into green fluorescent protein (GFP) eukaryotic expression vector pEGFP-C1 to construct recombinant pGFP-XB and pGFP-XC. The pEGFP-C1, pGFP-XB and pGFP-XC were introduced into Bel-7402 cells by Fugene HD to obtain Bel-7402 cells expressing GFP. The transcription and expression of HBx gene were demonstrated by RT-PCR and Western Blot analysis. Bel-7402, Bel-7402/GFP, Bel-7402/GFP-XB, Bel-7402/GFP-XC cells were treated with adriamycin (2.5 microg/ml), and the apoptosis of the cells was determined by trypan blue exclusion, and flow cytometry analysis.

RESULTSRT-PCR and Western Blot analysis showed that HBx genes of genotypes B and C were transcribed and expressed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Trypan blue exclusion showed adriamycin induced time-dependent cell death in Bel-7402, Bel-7402/GFP cells while no significant cell death was observed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Flow cytometry analysis indicated that no significant differences of apoptosis rates of Bel-7402/GFP-XB (3.87%) and of Bel7402/GFP-XC (4.01%) were observed (P > 0.05), moreover, no significant differences of Bel-7402/ GFP-XB (3.87%), Be17402/GFP-XC (4.01%) and of the untreated cells. Apoptosis rates in Bel-7402/GFP-XB (3.87%), Bel-7402/GFP-XC (4.01%) cells were significantly lower than those in Bel-7402 (27.05%) and Bel-7402/GFP (29.14%) cells at 48 hours after the adriamycin treatment (P < 0.01).

CONCLUSIONSBel-7402 cell lines expressing GFP, GFP-XB and GFP-XC fusion proteins were successfully established. HBV X protein blocks adriamycin-induced apoptosis of Bel-7402 cells. There is no difference between HBx of genotype B and C in inhibiting apoptosis induced by adriamycin.

Antiviral Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Hepatitis B ; drug therapy ; physiopathology ; virology ; Hepatitis B virus ; classification ; drug effects ; genetics ; metabolism ; Humans ; Trans-Activators ; genetics ; metabolism

Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; pathology ; surgery ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; diagnostic imaging ; pathology ; surgery ; Male ; Middle Aged ; Tomography, X-Ray Computed

Objective To investigate the mechanism of IL-17, the signature cytokine produced by Th17 cells, in OLP lesion. Methods 24 patients with reticular OLP, 19 patients with atrophic-erosive OLP and 13 healthy volunteers were enrolled in this study . Real-time quantitative PCR ( real-time qPCR ) was performed to analyze the expressions of the production of IL-17 and CCL20 mRNA. Results The expressions of IL-17 mRNA in reticular OLP and atrophic-erosive OLP were significant higher than that in healthy oral mucosa (P = 0.0095, P <0.0001, respectively), meanwhile, remarkable increased IL-17 expression in atrophic-erosive OLP group was found compared with reticular OLP group (P = 0.0012). Additionally, the expressions of CCL20 mRNA in reticular OLP and atrophic-erosive OLP were significant higher than that in control group (P=0.0357, P<0.0001, respectively), meanwhile, CCL20 expression in atrophic-erosive OLP was higher than that in reticular OLP. The expressions of CCL20 mRNA rises with the increased expression of IL-17, and were positive correlated with IL-17 expressions in OLP lesions (P=0.003). Conclusions IL-17 production can induce chemokine CCL20 expression in OLP lesion. The signal pathway may promote the migration and infiltration of inflammatory cells in OLP lesions.

BACKGROUND:Conventional surgical repair can cause large traumas in patients with knee injuries, and patients often recover slowly after implant fixation, most of whom can appear to have poor recovery of knee function. OBJECTIVE: To explore the folow-up effect of arthroscopic double-steel wire clip fixation on tibial eminence avulsion fracture of anterior cruciate ligament. METHODS: A retrospective analysis was performed on the clinical data of 23 patients with tibial eminence avulsion fractures, who were given arthroscopic double-steel wire clip fixation. The patients were folowed up for 1-6 months. Short- and middle-term therapeutic effect as wel as IKDC and Lysholm scores before and after treatment were observed and analyzed. RESULTS AND CONCLUSION:The operation time was 35-65 minutes, and no complications, such as blood, nerve and anterior cruciate ligament injuries occurred. Moreover, no infection and other poor biocompatible reactions occurred after internation fixation. Al patients were folowed up for 1-6 months. The excelent and good rate was 87% at 1 month after treatment and 96% at 6 months after treatment. Al the patients had improved IKDC score and Lysholm score after treatment (P < 0.05), indicating that the knee function of patients was improved significantly.

Objective To investigate the effects of fluoride at different concentrations on proliferation and apoptosis of primary rat ameloblast in vitro.Methods Ameloblasts were isolated from tooth germ of 4 days SD rat maxillomandibular molar and cultured in vitro.Cells were treated with NaF at 0.0 (control group),0.4,0.8,1.6,3.2 and 6.4 mmol/L for 24,48 and 72 h,respectively.Inverted microscope was used to observe cell morphology;immunochemistry method was used to identify ameloblasts;3-(4,5-dimethylthiazole-2)-2,5-diphenyl tetrazolium bromide (MTT) assay was applied to measure cell viability at each time point.The cells were treated with 1.6 mmol/L NaF for 24 and 48 h,or after 50 mol/L caspase pan-inhibitor Z-VAD-FMK pretreatment 1 h,1.6 mmol/L NaF treatment for 48 h.Cell apoptosis was then tested by flow cytometry.In addition,activation of caspase-3 and poly (ADP-ribose) polymerase (PARP) were assessed by Western blotting to explore potential involvement of caspase activation in NaF-induced apoptosis.All data analysis was performed using SigmaStat V 3.5 software.Results ①Primary rat ameloblasts were in polygonal shape at low density and appeared like paving stone at high density with obvious nucleus,showing typical morphological characteristics of cells with epithelial origin.②The results of immunochemistry assay indicated that the cultured cells were positive in cytokeratin 14 (CK14) and ameloblastin (AMBN) staining,in accordance with the immunocytochemical characteristics of ameloblasts.③The effects of NaF on ameloblast proliferation were in a dose-and time-dependent manner.For low dose NaF (0.4 and 0.8 mmol/L) groups,cells treated for 24 h had significantly higher cell proliferation rates than that of the control group (0.0 mmol/L,P ＜0.05),the proliferation indexes for 0.0,0.4 and 0.8 mmol/L groups were 1.00 ± 0.00,1.38 ± 0.11 and 1.29 ± 0.13,respectively;the same doses of NaF had no obvious influence on cell proliferation at 48 h (1.00 ± 0.00,1.16 ± 0.14 and 0.94 ± 0.07,P ＞ 0.05);cell proliferation indexes at 72 h were significantly lower than that of the control group (0.87 ± 0.03 and 0.80 ± 0.04,P ＜ 0.05).Medium dose of NaF (1.6 mmol/L) did not cause obvious alterations in cell proliferation at 24 h (0.90 ± 0.08,P ＞ 0.05);while cell proliferation indexes at 48 and 72 h were obviously reduced than that of the control group (0.38 ± 0.03 and 0.26 ± 0.04,P ＜ 0.01).For high NaF concentration (3.2 and 6.4 mmo]/L) groups,cell proliferation indexes were significantly decreased at all time points compared with control cells,the rates for 3.2 mmol/L groups were 0.57 ± 0.14,0.08 ± 0.03 and 0.00 ± 0.00,respectively,and the rates for 6.4 mmol/L groups were 0.11 ± 0.04,0.00 ± 0.00 and 0.00 ± 0.00,respectively (P ＜ 0.01).④Flow cytometry was used to detect apoptosis.The results showed that treatment with 1.6 mmol/L NaF resulted in significantly increased apoptosis in ameloblasts at both 24 h [(5.80 ± 2.03)％] and 48 h [(17.45 ± 4.97)％] compared to the control group [(2.59 ± 0.95)％,P ＜ 0.05].In cells pre-treated with pan-caspase inhibitor Z-VAD-FMK,NaF-induced apoptosis was significantly lower than that of cells treated with only 1.6 mmol/L NaF [(9.43 ± 3.79)％ vs (18.26 ± 3.39)％,P ＜ 0.05].⑤Cleavage of caspase-3 and PARP was detected in ameloblasts treated with 1.6 mmol/L NaF for 48 h.Conclusion Overdose fluoride could inhibit proliferation and induce apoptosis via activation of caspase cascade in primary cultured rat ameloblasts.

Objective:To investigate the polymorphism of HLA-DQA1 genes in Jing nationality of Central Vietnam.Methods:Applied PCR-SSP tecnique to determine the polymorphism of the HLA-DQA1 alleles of 105 healthy children and youth,unrelated individuals in Central of Vietnam.Results:10 HLA-DQA1 alleles were detected of which DQA1*0104 were the most common allele with frequency of 21.3% and lowest frequency is DQA1*0601.Conclusion:The results indicate that HLA-DQA1 alleles polymorphism of Jing nationality in Central Vietnam is different from the other Chinese. [

OBJECTIVETo investigate midterm follow-up results on Asian femoral intramedullary nail in treating segmental and comminuted femoral fractures.

METHODSBetween June 2011 and October 2012,16 patients with segmental and comminuted femoral fractures were treated with minimally invasive reset and Asian femoral intramedullary nail under extension table. Among them, there were 10 males and 6 females aged from 21 to 49 years old with an average of 34.5 years old; the time from injury to operation ranged from 3 to 24 d with an average of 9.1 d. There were 6 cases were type C1,2 cases were type C2 and 8 cases were type C3 according to AO classification. X-ray of femoral segment at 3,6 and 12 months after operation were applied for evaluating fracture healing. Harris score of hip joint and HSS score of knee joint were used to evaluate postoperative function.

RESULTSAll patients were followed up from 24 to 36 months with an average of 28.4 months. Operative time was from 88 to 112 min with an average of 90.7 min; blood loss ranged from 150 to 200 ml with an average of 188.75 ml; the time of fracture healing was from 5 to 9 months with an average of 5.4 months. All incision were healed at stage I. No loosening, breakage of internal fixation and displacement of fracture were occurred. There were no significant differences in Harris score of hip joint at 3, 6 and 12 months after operation (F = 0.07, P = 0.893 > 0.05), 10 cases obtained excellent results, 5 good and 1 moderate. There was no obvious meaning in HSS score of knee joint (F = 0.08,P = 0.876 > 0.05), 9 cases obtained excellent results, 6 good and 1 poor.

CONCLUSIONAsian femoral intramedullary nail could treat segmental and comminuted femoral fractures by using variety of less invasive ways,which has advantages of less trauma, quick recovery of function and satisfied midterm following-up results. But long term following-up effects remains to be seen.

Adult ; Bone Nails ; Female ; Femoral Fractures ; surgery ; Femur ; injuries ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; Fractures, Comminuted ; surgery ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

ObjectiveTo establish the time-resolved fluoroimmunoassay (TRFIA) method for the detection of serum galectin-3 and investigate the clinical value of serum galectin-3 for the diagnosis of pancreas cancer.MethodsMonoclonal anti-human galectin - 3 antibody and biotinylated polyclonal antibody were used to establish the sandwich TRFIA for detection of serum galectin-3.The optimal experimental condition was studied.Serum levels of galectin-3,CEA and CA19-9 in the patients with pancreatic cancer,benign pancreatic mass,pancreatitis,and healthy controls were measured.The diagnostic value of serum galectin-3,CEA and CA19-9 for pancreas cancer was studied.ResultsThe linearity of the TRFIA for detection of serum galectin3 tanged between 0 to 100 μg/L.The within-run CV and between-run CV were ≤6.45％ and ≤8.68％,respectively,and the average recovery was 106.6％.The level of serum galectin-3 was 4.93 ( 0.85 ～ 23.80) μg/L in pancreatic cancer group,which were significantly higher than those in benign pancreatic mass [2.83 ( 2.17 ～ 4.06) μg/L ],pancreatitis [ 2.62 (0.55 ～ 9.76 ) μg/L ],and healthy controls group [ 1.88 ( 0.59 ～ 3.94) μg/L] (P ＜0.05).By using 3.77 μg/L as the cut-off point,the smsitivity,specificity for the diagnosis of pancreatic cancer was 75.5％ and 90.9％.The levels of Gal 3 and CEA,CA19-9 was not correlated ( r =0.1321,P =0.3761 ; r =0.0920,P =0.5384).Combined determination of galactin-3 and CEA,CA19-9 levels could increase the diagnostic sensitivity to 92％.ConclusionsTRFIA method for the detection of galactin-3 is sensitive and stable.Galectin-3 could be a potentially novel serum tumor marker of pancreatic cancer.

Objective To evaluate the clinical outcome and the acute toxicity in nasopharyngeal carcinoma (NPC) treated with tomotherapy followed by the anti-EGFR monoclonal antibody.Methods Between March 2008 and November 2009,34 newly diagnosed NPC patients were treated with helical tomotherapy combined with nimotuzumab or cetuximab.All the patients underwent tomotherapy at the dose of 70 Gy/33F for the gross tumor volume (pGTVns) and positive lymphnodes (GTVnd) ,and 60 Gy/33F for the high risk clinical target volume (PTV1),and 56 Gy/33 F for the low risk clinical target volume (PTV2),respectively.17 patients in group N were given weekly injection of 200 mg for 6-7 times and 17 patients in group C were given initial dosage 400 mg/m2 followed by subsequent weekly dosage of 250 mg/m2 for 6-7 times.Acute lesions were evaluated with the RTOG/EORTC criteria.Result The median follow-up time was 22 months.The effective rates (CR + PR) in 3,6 and 12 months were 14/17,12/17,12/17 in group N and 15/17,14/17,14/17 in group C.The 1 year survival rate was 15/17 in group Nand 17/17 in group C.Nimotuzumab had less acute mucositis reaction (u = 2.25,P < 0.05),weight loss(t=2.56,P=0.02) and rash (u=4.36,P＜0.01) compared with cetuximab.Conclusions Helical tomotherapy combined with nimotuzumab or cetuximab was effective and made no difference in the shortterm efficacy and 1 year survival rate for the patients with NPC.Nimotuzumab has less acute reaction than cetuximab.More studies should be done to prove long-term effects.

ObjectiveTo explore the role of Network Management System (NMS) in decreasing mortality and incidence of cerebral palsy in preterm infants.MethodsThe data of 356 preterm infants transported by NMS from January 2004 to December 2005 were analyzed.ResultsNo death cases occurred during the transportation of 356 preterm infants, the success rate was 100%. 292 cases (84.39%) were cured and 36 cases (10.4%) were effective. 7 case dead for compliance, the mortality was 19.6‰. 3 cases suffered from cerebral palsy , the incidence of cerebral palsy was 8.6‰.ConclusionNMS applied to preterm infants is a high-effective medical model, and plays an important role in improving the forward prognosis of preterm infants.