Arginine Deiminase(ADI) was purified to homogeneity using ammonium sulfate precipitation,Q-Sepharose Fast Flow anion exchange chromatography and SephadexG-75 gel filtration chromatography. This purification protocol resulted in a 34.5-fold purification of ADI with 31.4% final yield. A molecular weight of about 190 kD determined by native gradient polyacrylamide gel electrophoresis. The enzyme has only one kind of 46 kD subunit determined by SDS-PAGE. Combining the results from the two kinds of electrophoresis,the authors deduce that the enzyme may be a tetramer. The optimum pH and temperature for lipolytic activity of ADI was pH 6.5 and 50℃,respectively. It was extremely stable at 45℃ and retained 97.9% of its original activity for 30 min. The stability declined rapidly as soon as the temperature rose over 50℃. ADI was highly stable in the pH range from pH 5-8. ADI acted on L-arginine but not on D-arginine. ADI catabolism was dependent on metal ions. At their adequate concentration,Mn2+,Mg2+ and Co2+ were the effective promoter,while superfluous Zn2+and Co2+ inhibited ADI activity. L-citrulline did not act on ADI,but L-ornithine inhibited ADI activity. The degradation of L-arginine with ADI catalysis was according to simple Michaelis-Menten equation. The Michaelis constant was 3.2686 mmol/L and the maxi-mum velocity was 2.44 ?mol/min.

L-theanine (γ-glutamylethylamide) is the main free amino acid component of tea and its favorable physiological effects on mammals have been reported. An enzymatic method for optically pure L-theanine production with a new L-aminoacylases-production fungi Cunnighamella echinulata 9980 was developed. The optimum conditions for enzymatic catalysis were at pH 6.5 with 50 mmol/L N-Acyl-DLtheanine and 40 mmol/L CoCl2. After 12-h incubation at 50℃,22.5 mmol/L L-theanine was obtained, the conversion rate against N-Acyl-L-theanine being 90%. Cunnighamella echinulata and the aminoacylase were applied in preparation of L-theanine.

OBJECTIVETo investigate the way of diagnosis and therapy of parosteal osteosarcoma.

METHODSA retrospective review was conducted of 48 patients treated at our department between June 1964 and December 2001. The average age of patients in this study was 29.2 years (13 - 47 years). Thirty-two of the patients were female; sixteen were male. The single most common site is the posterior aspect of the distal femur (in 36 patients), followed by the femur shaft (in 6 patients), the proximal tibia (in 2 patients), etc. Nine patients had been operated on before referral to our department.

RESULTSThe average follow-up period from the first operation was 5.2 (0.25 - 24) years. In 36 of the 39 patients in whom a limb-salvage procedure was performed, a segment of the tumor-bearing bone was excised along with the tumor, whereas in 3 patients only the subadjacent cortex was excised with the tumor. In the limb-sparing group, the reconstruction was achieved by means of attenuated tumor bone or allograft in 23 cases, by endoprosthetic replacement in 9 cases, and by allograft replacement in 4 cases. The local resections were wide in 35 cases, and marginal in 13 cases. After marginal surgery, local recurrence occurred in 5/13 patients, whereas it occurred in 3/35 patients treated with wide resection. Pulmonary metastases developed in 6 patients, four patients died, and 2 patients are alive with disease. There were 4 cases of fractures of bone grafts. Four patients developed an infection. Long-term survival rate is 85.8%. For tumors that invaded the medullary canal there was no statistical association with local recurrence or metastasis. There is statistical significance between surgical margin and local recurrence.

CONCLUSIONSWide surgical excision alone is adequate treatment for patients with conventional parosteal osteosarcoma. A tumor-free margin remains the critical factor determining overall prognosis. When a marginal excision was knowingly done to preserve a major neurovascular bundle, the risk of recurrence was less than when it was done to shell-out a presumptively benign lesion. Repeated recurrence probably increases the risk of dedifferentiation and thereby worsens the prognosis. Recurrent lesions with multiple soft-tissue satellite nodules or involvement of the neurovascular structures may however require amputation to provide sufficient local control when a wide margin cannot be achieved. An individualized resection will be performed in the future probably under the help of the advanced technique of image to distinguish the reactive zone from the normal tissue precisely.

Adolescent ; Adult ; Bone Neoplasms ; diagnostic imaging ; surgery ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Osteosarcoma, Juxtacortical ; diagnostic imaging ; surgery ; Prognosis ; Radiography

Background: The side effects of glucocorticoid in treatment of systemic lupus erythematosus (SLE) have been the focus of debate, and our preliminary study indicates that ginsenosides can enhance the efficacy of dexamethasone. Objective: To observe the effects of ginsenosides combined with prednisone in SLE patients. Design, setting, participants and interventions: A total of 60 SLE patients from Department of Rheumatology and Immunology, Changhai Hospital, Second Military Medical University, were randomly divided into treatment group and control group, with 30 patients in each group. Patients in the treatment group were given routine treatment with prednisone plus ginsenosides, while those in the control group were given routine treatment with prednisone plus placebo. They were all treated for 3 months. Main outcome measures: After three-month treatment, syndrome score in traditional Chinese medicine (TCM), total response rate and symptom improvement rate were measured and evaluated. Results: Twenty-eight cases in treatment group and twenty-seven cases in control group were included in analysis. The total response rates in the treatment group and control group were 89.28% and 66.67% respectively, and there was a significant difference between the two groups (P<0.05). After treatment, the TCM syndrome scores in the two groups were lower than those before treatment (P<0.01), and prednisone plus ginsenosides was better in decreasing the TCM syndrome score than prednisone plus placebo (P<0.05). The symptoms were improved in the treatment group as compared with the control group (P<0.05). Conclusion: Prednisone combined with ginsenosides can increase the clinical effective rate and improve the clinical symptoms of SLE patients.

A bacterial strain DN25, effective on cyanide-degradation, was isolated from contaminated soil and identified as Alcaligenes sp. on the basis of phenotype analysis and 16S rDNA sequence analysis. It showed great tolerance to the cyanide, which can grow in the medium containing 500mg CN -/L. The suitable condition for the cell growth and boitransformation was pH8.0 and 30oC and the transformation rate for 500mg CN - /L could achieve 99% in 10 h. It has also been found that the screened strain had the ability of K 4Fe(CN) 6 transformation with 96% of transformation rate at 12 h for the concentration of 500 mg CN /L.

Objective To observe the relationship between apoptosis of K562 cell induced by iron-deprivation and activation of Caspase-3.Methods K562 cells were treated with desferrioxamine(DFO) in different dosages were collected at different time points.K562 cells were labelled with Annexin V/PI,and then the rate of apoptosis was measured by flow cytometry;The activation of Caspase-3 were detected by colorimetric method with pAN labelled substrate;The active protein of Caspase-3 were analyzed by Western blot.Results When K562 cells were treated with different concentrations of DFO,the apoptosis rate and the activity of Caspase-3 increases gradually.When K562 cells were incubated with DFO(50 ?mol/L and 100 ?mol/L) 24 h later,the enzymatic activity of Caspase-3 increases dramatically more than that of control group,and the difference was significantly(P0.05).All those effect above can be counteracted by equal mole concentration of FeCl_3.Conclusion Iron-deprivation maybe induce the apoptosis of K562 cell by chelating intracellular iron and activing Caspase-3.

OBJECTIVETo evaluate the differences in selecting patients who will undergo operation by comparing the evaluating systems and to discuss the indication of surgery performed on patients with spinal metastases.

METHODSOf the 113 patients presented at Jishuitan Hospital, Beijing, PRC. from February 1994 to February 2002, who were diagnosed as spinal metastases, the clinical, pathological and imaging records of 82 patients were reviewed. Harrington's classification system, Tokuhashi's scoring system, Tomita's scoring system, and Grubb's scoring system were used to evaluate each of these patients at the same time. The classification or score concerning each patient was recorded and compared. SPSS software was used for statistical analysis. McNemar Test was used to compare the proportion of surgery indicated by different evaluating systems.

RESULTS(1) Surgical treatment was indicated for 92.7% of patients by Tomita's scoring system and 69.5% by Harrington's classification system respectively. The difference was statistically significant at the level of P < 0.001. (2) Seventy-five cases with spinal cord palsy and progressive pain could be assessed by Tokuhashi's system. These patients were evaluated by Tomita's system at the same time. The type of surgery determined by Tokuhashi's score was excisional operation for 20% of the patients, palliative operation for 20%. As for the remaining 60%, either excisional or palliative operation might be chosen. The type of surgery determined by Tomita's score was excisional operation for 49.3% of the patients, palliative for 42.7%, and nonsurgical modality for the remaining 8.0%. (3) Twenty-five patients were classified as I, II or III, by Harrington's classification system. These patients were evaluated by the Grubb scoring system to predict the possibility of pathologic fracture and by Tomita's system at the same time to determine if an operation was necessary. By Grubb's system, prophylactic stabilization was necessary for twenty-two patients. By Tomita's system, surgery was chosen for 23 patients. But the treatment options were different in 5 patients assessed by the two systems.

CONCLUSIONSThere was a great difference in the selection of treatment modality for patients with spinal metastases by different evaluating systems. That no systems have incorporated the evaluation of life expectancy and the local lesion causes differences. The patients should be evaluated individually by more factors than those proposed by the established systems.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Preoperative Care ; methods ; Prognosis ; Proportional Hazards Models ; Spinal Neoplasms ; diagnosis ; secondary ; Survival Analysis ; Young Adult

OBJECTIVETo improve the life quality of cancer patients with metastasis to long bone and to select suitable surgical treatment.

METHODSFifty two patients with metastasis 27 men and 25 women, were treated from 1990 to 1999. Their average age was 56.8 years (33 - 74). In 16 patients with multiple lesions, underwent surgery at bone shaft (29 patients) and bone epiphysis (26). Thirty patients were treated for pathologic fracture and the rest for impending fracture. Operations included limb-salvage (51 patients) and amputation (4) Limb salvage consisted of intralesional curettage (3 patients), intramedullary nailing reconstruction (29), endoprosthesis (18), and temporary spacer (1). 21 patients accepted postoperative chemotherapy or radiotherapy.

RESULTSFollow-up of 52 patients for a mean of 28.2 months (2 - 122 months) showed pain relief (41 patients), (75%) and full or part weight-bearing stability (36) 69%. Local tumor recurrence occurred in 11 patients.

CONCLUSIONSSurgical treatment can effectively improve the life quality of patients with metastasis to long bone. The metastatic lesions should be resected with wide or radical margin for the patients with kidney, breast, prostate and thyroid cancer.

Adult ; Aged ; Bone Neoplasms ; pathology ; secondary ; surgery ; Female ; Follow-Up Studies ; Humans ; Limb Salvage ; Male ; Middle Aged ; Treatment Outcome

Animals ; Escin ; pharmacology ; Intestines ; blood supply ; Lipid Peroxidation ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; prevention & control

OBJECTIVETo study the molecular mechanism of apoptosis of leukemic cells (K562 cells) induced by iron chelating agent deferoxamine (DFO).

METHODSThe exponentially growing K562 cells were used (1×10(6)/mL) in this study. The K562 cells were treated with different concentrations of DFO (10, 50 and 100 mmol/L), DFO+FeCl3 (10 μmol/L each) or normal saline (blank control). The cellular labile iron pool was measured with a fluorimetric assay using the metalsensitive probe calcein-AM. The viable count and cell viability were determined by typanblue assay. Cell apoptosis was determined by morphological study and flow cytometry assay. Caspase-3 activity in K562 cells was detected by colorimetry.

RESULTSAfter DFO treatment, the cellular labile iron pool and the viability of K562 cells were reduced and the cell apoptosis increased in a time- and dose-dependent manner compared with the blank control group. The apoptosis rate of K562 cells in the DFO+FeCl3 treatment group was not significantly different from that in the blank control group. The caspase-3 activity in K562 cells increased significantly 24 hrs after 50 and 100 μmmol DFO treatment when compared with the blank control group (P<0.01). There was a negative correlation between cellular labile iron pool and caspase-3 activity of K562 cells (r=-0.894, P<0.05).

CONCLUSIONSDFO induces apoptosis of leukemic cells possibly through decreasing cellular labile iron pool and increasing caspase-3 activity of the cells.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Deferoxamine ; pharmacology ; Flow Cytometry ; Humans ; Iron Chelating Agents ; pharmacology ; K562 Cells