BACKGROUND: A process of -80 ℃ profound hypothermia over three months and freeze drying will suppress the histocompatibility antigen of decalcified freeze-dried bone and reduce their immunogenicity. OBJECTIVE: To investigate the effect of treating intraarticular fracture using allogeneic decalcified freeze-dried bone nail alone. DESIGN, TIME AND SETTING: From August 2000 to August 2007, a clinical analysis was performed among 83 patients of cancellous bone fracture involving articulus, who were enrolled from Department of Orthopaedics in the Central People's Hospital of Zhanjiang (Zhanjiang, Guangdong, China). MATERIALS: The involved patients consisted of 64 males and 19 females, aged 16-53 years with a mean of 36.3. The lesioned sites were elbow joint in 28 cases, acetabulum in 6 cases, femoral condyles in 9 cases, tibial plateau in 17 cases and ankle joint in 23 cases. Allogeneic decalcified freeze-dried bone nail was developed by the Biomaterial Laboratory of the Southern Medical University (also known as the First Medical University of Chinese PLA). METHODS: Totally 83 cases of cancellous bone fracture involving articulus were fixed with bone nail or bone screw after reducing of bone fracture. Bone nails or screws were pretreated with saline or allogeneic blood over 30 minutes. Bone nail at 3.5-mm diameter were applied on elbow joints and ankle joints, while those at 4.0-mm diameter for knee joints and hip joints. After implantation, hip and femoral condyles fractures were treated with the traction for 4-6 weeks, and the fractures of elbow joints, ankle joints and tibial plateau were fixed with gypsum for 4-6 weeks. Patients received functional exercise after wound healing. Subsequent to draining for 48 hours, antibiotics were used to prevent infection, but immune depressant was unnecessary. MAIN OUTCOME MEASURES: Wound healing was observed. The growth of osteotylus and the absorption of bone nail were checked with X-ray. Immunological rejection was also checked. RESULTS: One case had dystopia at postoperative 2 weeks, one case with breakage of bone nail was found at postoperative 3 weeks, and two cases developed immunological rejection in advanced stage. The other 79 cases got bone healing without dystopia or immunological rejection, and they achieved bone union within 1-3 months with normal joint function. CONCLUSION: Being internal fixation material of intraarticular fracture, allogeneic bone nail can maintain the stabilization of cancellous bone fracture to bone healing, exhibit complete absorption and be good at bone repair.

Objective To study the effect of radiofrequency ablation (RFA) with sublethal temperature on the production of liver cancer stem cells (LCSCs) and on the expression of LCSCs-related transcriptional factors.Methods Mouse hepl-6 hepatoma cell line and clinical samples of patients with hepatocellular carcinoma (HCC) were used to test the expressions of LCSCs-related markers and transcriptional factors.Results Different temperatures were used to stimulate Hep1-6 cells,and it was proved that the temperature of 45℃ was a sublethal temperature that could not induce cell death.Flow cytometry testing showed that treatment with 45℃ could obviously increase CD13+,CD44+,CD90 and CD133+ Hep1-6 cells,suggesting that treatment with 45℃ could increase the production of above mentioned types of LCSCs in hep1-6 cells.Real-time quantitative polymerase chain reaction (RT-qPCR) assay indicated that the temperature of 45℃could cause significant increase in CD13,CD90 and CD133 mRNA.In all 5 HCC patients,CD13 mRNA in the recurrent HCC lesions was remarkably increased,CD133 mRNA was increased in 4 patients with recurrent HCC,and CD90 mRNA was increased in only one patient with recurrent HCC.Flow cytometry testing revealed that CD13+ LCSCs were strikingly increased in 4 recurrent HCC patients,while CD133+LCSC was increased in only one patient,suggesting that more close correlation existed between the increase of CD13+ LCSCs and the temperature of 45℃.RT-qPCR assay showed that in 4 recurrent HCC patients with increased CD13+ LCSC,the Sox2 and Stat2 among 13 LCSCs-related transcriptional factors were obviously increased.Flow cytometry testing showed that 45℃ treatment also increased the expression of Sox2 and Stat1 mRNA in Hep1-6 cells.Finally,Sox2 and Stat1 could be knockdown by siRNAs,indicating that both Sox2 and Stat1 transcriptional factors were involved in 45℃-induced production of CD13+ LCSCs in Hep1-6 cells.Conclusion In RFA therapy,the use of sublethal temperature of 45℃ can increase CD13+LCSCs,which is related to the promotion of Sox2 and Stat1 expression.The results of this study can be used for reference in the research of liver cancer recurrence.

Objective: To investigate the genetic characteristics of Lamivudine-resistant mutation patterns and HBV S gene mutants in patients with chronic hepatitis disease of different disease progression. Methods: Blood samples of LAM-resistant patients with chronic hepatitis disease were collected. HBV RT gene nucleotide sequences were obtained, and then differences in drug-resistant mutation patterns, drug susceptibility and HBV S gene mutants characteristics between the two groups were analyzed. Results: Forty-seven chronic hepatitis B (CHB) patients and 16 HBV-related liver cirrhosis (LC)/HBV-related hepatocellular carcinoma (HCC) patients were included in this study. M204I single point mutation and L180M+ M204I/V were the most common pattern during patients with chronic hepatitis disease (35/63, 55.56%). The numbers of resistant to three nucleos(t)ide analogues in LC/HCC group was higher than CHB group’s (62.50% vs 34.04%, P=0.046). In HBV S gene, more immune associated HBsAg-escape mutations were detected in LC/HCC group than that in CHB group (62.50% vs 31.91%, P=0.031). I126T/V and G145A (for LCC/HCC group, 60%), I126S/T and S117T (for CHB group, 46.67%) were showed as the most common form for HBsAg escape mutations in the two groups. The two groups both detected RT mutations concomitantly with stop codon mutations in S gene (rtA181T/sW172* and rtM204I/sW196*). Conclusions Different characteristics in Lamivudine-resistant mutations and associated HBV S gene mutants were found in patients with chronic hepatitis disease of different disease progression, and LC/HCC patients exhibit more multi-drug resistant variants and immune associated HBsAg-escape mutants than CHB patients.