Objective To discuss the treatment and prognosis of multifocal osteosarcoma. Methods Between January 1998 and June 2005, nine patients(seven males and two females), six with synchronous multifocal osteosarcoma and three with metachronous multifocal osteosarcoma were treated in our department. The diagnosis of multifocal osteosarcoma was confirmed by clinical presentation, radiology and pathology. Eight of the nine patients received chemotherapy with adriamycin(60 mg/m2), cisplatinum(100 mg/m2), high-dose methotrexate(8-12 g/m2), vincristine(1.4 mg/m2), ifosfamide(12.5 g/m2), and G-CSF. Based on the response to primary chemotherapy, further chemotherapy plans were made, and different operations were done to control local lesions or to relieve symptoms. Arsenous acid, paclitaxel and VP-16 were used in patients who responded poorly to primary chemotherapy. Results One patient who given up remedy was died in systematic fail due to the cancer after 5 month, five patients who were diagnosed synchronous multifocal osteosarcoma died from progressive disease in 3.5 to 17 months (mean time was 9.8 months). Only two of them had a good response to chemotherapy and had finished at least four cycles of chemotherapy,and they had a better survival time of 15 and 17 months respectively. Three patients with metachronous multifocal osteosarcoma had a better prognosis. After a two-year follow-up, two lived with disease and one lived without disease. Conclusion The prognosis of patients with multifocal osteosarcoma is poor, and patients with metachronous multifocal osteosarcoma have a relatively better prognosis. To improve the prognosis of patients with multifocal osteosarcoma, aggressive chemother-apeutics are needed.

[Objective]To evaluate the effectiveness of biopsy of bone lesions.The factors that might effect on the clinical utility were analyzed in order to find out the independent factor for clinical effectiveness.Also we evaluated the effects of these factors on the open biopsy with needle biopsy previously done.[Method]We retrospectively reviewed 206 cases performed at our institute between June 2003 and Oct 2006,including percutaneous core-needle biopsy or open biopsy.The biopsy result was classified as correct,nondiagnostic,or incorrect.The biopsy results were considered clinical useful if they were correct and not useful if they were nondiagnostic or incorrect.We identified the independent risk factor by analyzing whether the characteristics including biopsy methods,lesion position,myxoid,lesion type reduced the effect on the clinical utility and increased the probability of proceeding to subsequent open biopsy.[Result]In 206 patients,one hundred and forty-seven cases(71.4%)underwent core-needle biopsy.Forty-seven cases(22.8%)underwent open biopsy and twelve cases(5.8%)had open biopsy with needle biopsy first.One hundred and seventy-six cases(85.4%)patients had clinical useful results.Eighteen cases(8.7%)were nondiagnostic,and twelve cases(5.8%)were incorrect.Myxoid histology was identified of independent risk factor on the clinical utility by logistic regression(P=0.02).[Conclusion]Most cases of bone lesions can be diagnosed by core-needle biopsy or open biopsy.The myxoid histology is associated with decrease clinical accuracy;No single characteristic cause increase probability of open biopsy.Correct diagnose of bone lesions requires combining biopsy,radiology and clinical data.

Objective To investigate the clinical manifestation and diagnosis of osteitis deformans. Methods Three male patients with characteristic manifestations of osteitis deformans were reviewed in this study. The ages of the patients were 51, 49 and 43 respectively. Two patients with pelvic lesions on X ray films presented symptomatic pain and malaise localized in hip, one complained of tibial bowing deformity. The typical radiographic characteristics were bone enlarged size, increased bone mineral density with uneven distribution; the bone trabeculae were usually coarse and arrayed irregularly like a grid. All patients underwent the diagnostic biopsy. Under microscopic observation, the bone structures were arranged disorderly, the trabeculae were thickened and enriched with osteoclasts and osteoblasts, irregular blue cement line were also seen in thickened trabeculae, the bone marrow were replaced by the fibrovascular connective tissue. All patients were given calcitonin and diphosphonates as medical treament. Results In the follow up period of an average of 16 months(range 3-26 months), one patients had pain relief at lesioned hip but functional limitation remained, the others did not have any sign of malignant progression. The serum alkaline decreased dramatically and the involved bone did not present with deteriorated change. They were working as usual. Conclusion Osteitis deformans is a disorder of bone architecture resulting from disturbance in the rate of bone turnover; typical radiographic and pathologic feature is the important points for definite diagnosis of osteitis deformans.

SUMMARY To describe the first case of leiomyosarcoma of the tibia with multiple skeletal metastases in China. A 31-year-old woman was referred to Peking University People’s Hospital for evaluation of a lesion on the proximal left tibia. The radiographic examination showed the lesion had a moth-eaten destructive appearance with ill-defined border and soft tissue extension. The tumor is isointense to muscle on T1-weighed images and heterogeneous and of high signal on T2-weighed images. Technetium-99m bone scintigraphy demonstrated multiple markedly increased skeletal radioisotope uptake. The tibia lesion was surgically removed. Microscopically, tumor cells consisted of spindle shaped cells, arranged in bundles, with cigar-shaped and blunt-ended nuclei. Immunohistochemistry revealed that the tumor cells were positive for smooth muscle antigen (SMA) and caldesmon. It should be cautious to make the diagnosis of leiomyosarcoma arising in bone. After excluding metastasizing leiomyosarcoma of gastrointestinal tract, urinary system and uterus, the final diagnosis relies on immunohistochemical analysis, however, other spindle cell sarcomas have to be differentially diagnosed.

Objective To evaluate the effect of extracorporeal membranous oxygenation (ECMO) on the small bowel preservation in a porcine model of controlled donors after cardiac death (CDCD).Method The domestic crossbred donor pigs were anesthetized and ventilated with 100％ oxyger.An intravenous cannula was placed through iliac arteries and jugular vein,and connected to ECMO system.The ECMO was performed to infuse abdominal organs when cardiac death was declared.Pathology,electron microscope,energy metabolism and cell apoptosis level of intestinal mucosa were evaluated before operation,in cardiac death and at the end of extracorporeal support,respectively.Result The normothermic extracorporeal support could quickly restore graft blood supply and oxygenation.One-h extracorporeal support could improve the energy status in intestine of donation after cardiac death (DCD).The histologic damage and apoptosis of 1-h extracorporeal support had no significant difference with those before operation and in cardiac death (P＞0.05).With the extension of extracorporeal support,the intestinal mucosa damage degree was gradually increased,and the content of adenosine triphosphate in intestinal mucosa reduced gradually.Conclusion The normothermic extracorporeal support for 1 h in DCD is beneficial for improving the viability of small bowel.However,the integrity of intestinal mucosa is destroyed gradually as extracorporeal supporting time over,which may be initiated by the activation of intestinal epithelial apoptosis.

Objective To investigat the changes of hemodynamics and arterial blood gas during the establishment of ECMO model and provide an ideal platform for the study of the mechanism of DCD protection using ECMO, a porcine model of controlled donors of cardiac death (CDCD) with extracorporeal support was established in the paper. Methods Eight domestic crossbred pigs were anes-thetized, atracurium (1mg/kg) and heparin (150~200 U/kg) was administered and the ventilator was discontinued while the animal was under deep anesthesia to kill the animal.Meanwhile, blood was drained from the left jugular vein catheter into the ECMO circuit and re-turned into the left femoral artery to establish the porcine model of CDCD with extracorporeal support . The average blood press and arterial blood gas were ministered at normal, cardiac arrest and ECMO supporting 1 h, 3 h and 5 h. Results The model of CDCD with extra-corporeal support was established successfully.Two animals restored the beating of the heart and the high doses of potassium chloride were given untill cardiac arrest during extracorporeal support.Blood gases throughout the apneic time showed a severe hypoxemia and acidosis, pH and PaO2 reduced to 7.09 ±0.24 and 5.3 ±0.93mmHg respective-ly, which can be improved during extracorporeal support.MAP and PaO2 were maintained over 300mmHg and 65mmHg respectively. Conclusion A new porcine model of controlled donors of cardiacdeath with extracorporeal support has been successfully established and showed to be feasible for related pathophysiological research work.

Objective To elucidate the genome organization of small deletion mutants of hepatitis B virus(HBV).Methods Amplified the HBV genomes by polymerase chain reaction from the serum of the patients with chronic hepatitis B and cloned the small HBV DNA less than 1 kb,then sequenced and analyzed the gene organization of these small deletion mutants of HBV.Results Totally one hundred and twenty-four small deletion mutants of HBV genomes categorized to sixty-four types were obtained and classified into three groups according to the criteria of the characteristics of gene organization,for example,spliced variants,regular deletion mutants and the deletion mutants with an internal poly (dA).All of these isolated mutants shared some common features as the deletion in coding regions and regulatory elements,66% of the mutants retained the cis elements crucial for the viral replication and encapsidation,while 48% retained the X region.Conclusions Small deletion mutants of HBV are commonly detected in the serum from chronic hepatitis B patients,the characteristic structure of such mutants implies that they might be closely co-related with the pathogenicity of HBV.The exact mechanisms need further study yet.

Objective To evaluate the clinical efficiency of humanized anti-CD52 monoclonal antibody (Campath-1H) and anti-CD25 monoclonal antibody (Zenapax) induction therapy in intestinal transplantation patients.Method The data of 6 patients receiving Campath-1H and 5 patients receving Zenapax induction therapy in intestinal transplantation between 2007 and 2012 were analyzed retrospectively.The counts of peripheral blood lymphocytes and monocytes,incidence of rejection and infention,and liver and kidney toxicity of recipients were recorded before and 3 months after transplantation.Results Of 6 intestinal transplantation patients receiving Campath-1H induction therapy,1 died of acute heart failure on the postoperative day 3,and the rest 5 patients had a powerful depletion of lymphocytes and monocytes in 8 weeks,followed by gradual increases after 8 weeks.The percentage of peripheral blood CD3 + T cells,CD4 + T cells,and CD8 + T cells was dropped to 5％ before administration,and remained at a steady low level in the first 8 weeks after induction.Of 5 patients receiving Zenapax induction therapy,1 died of Aspergillus infection on the postoperative day 25,and the rest 4 patients had an obeivous increase of lymphocytes and monocytes on the postoperative day 1.Counts of lymphocytes and monocytes kept steady at normal levels from the 1st to 12th week.One case of mild rejection was found in Campath-1H group.One case of mild,one moderate and one severe rejection were detected in Zenapax group.All rejections were successfully cured by prompt anti-rejection therapy.There were no significant difference in serum creatimine,urea nitrogen,alanine aminotransferase or total bilirubin after 3 months in comparison to preoperation.Conclusion Both Campath-1H induction therapy and Zenapax induction therapy successfully induce immune tolerance in patients with intestinal transplantation.Campath-1H seems to offer better immunosuppression against Zenapax during the first 3 months posttransplantation.

Objective To investigate the anti-IFN-α effects of the novel protein TSR'r' encodedby the 458 nt-1308 nt spliced variant of hepatitis B virus genome,and to determine its functional domaias.Methods the TSR'r' gene(originated from open reading frame of HBV DNA polymerase,T represents terminal protein region,S represents the Spacer region,R'represents the truncated reverse transcriptase region,and r'represents the truncated RNaseH region)of the 458 nt-1308 nt spliced variant of HBV genome and its deletants were amplified by PCR and were cloned into the pcDNA3.1/HisC vector.The recombinant vector was transfected into Huh7 hepatocytes individually by FuGENE6 transfection reagent,and the expression of the fusion protein was detected by Western blot.Huh7 hepatocytes were co-transfected with p6 16CAT and the recombinant vector harboring either TSR'r'or the related deletant,and treated with IFN-α 2a 48 h post transfection.After 24 h stimulating.the cells were lysed and the intracellular CAT value was calculated.All data were processed with One-way analysis of variance(ANOVA).Resuits Recombinant vectors harboring either the TSR'r'gene or related deletant were constructed successfully,and the fusion proteins were expressed well in Huh7 cells.When Huh7 hepatocytes were co-transfected with p6-16CAT and TSR'r' recombinant.the intracellular CAT values reduced gradually as paralleled with the increasing amount of TSR'r'recombinant.Furthermore,as compared with the empty vector,intracellular CAT values also decreased significantly when the Huh7 cells co-transfected with recombinant harboring TP plus Spacer regions,while any of the other deletants(harboring either TP or Spacer region or neither)showed no significant difference.Conclusion The novel protein encoded by the 458 nt-1308 nt spliced variant of hepatitis B virns genome suppressed the response of Huh7 hepatocytes to IFN-α.and the N-terminal TP plus Spacer region was the functional domain of the protein for anti-IFN-α effects.

The study aims to develop a unified method to determine seven phenolic acids (neochlorogenic acid, chlorogenic acid, 4-caffeoylquinic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C) contained in honeysuckle flower that is the monarch drug of all the eight Yinqiao Jiedu serial preparations using quantitative analysis of multi-components by single-marker (QAMS). Firstly, chlorogenic acid was used as a reference to get the average relative correction factors (RCFs) of the other phenolic acids in ratios to the reference; columns and instruments from different companies were used to validate the durability of the achieved RCFs in different levels of standard solutions; and honeysuckle flower extract was used as the reference substance to fix the positions of chromatographic peaks. Secondly, the contents of seven phenolic acids in eight different Yinqiao Jiedu serial preparations samples were calculated based on the RCFs durability. Finally, the quantitative results were compared between QAMS and the external standard (ES) method. The results have showed that the durability of the achieved RCFs is good (RSD during 0.80% - 2.56%), and there are no differences between the quantitative results of QAMS and ES (the relative average deviation < 0.93%). So it can be successfully used to the quantitative control of honeysuckle flower principally prescribed in Yinqiao Jiedu serial preparations.