Objective To develop an ELISA for detection of anti-dsDNA antibody by using plasmid DNA as antigen.Methods DNA in plasmid pBV220 for prokaryotic expression vector was purified by base-cleavage. The microtiterplates were pretreated by poly-L-lysine and coated by the plasmid DNA in a dilution of 1∶50 as antigen. An ELISA method for detection of serum anti-dsDNA antibodies was developed with HRP-SPA as enzyme-labeled marker.The IIF using crithidia lucilia as substrate was performed simultaneously for comparison. The serum samples from 64 patients with SLE, 8 with MCTD and 17 with RA were detected. Results The concentration of DNA was 1 54 g/L by UV spectrophotometer at wavelength of 260 nm. The positive percentage of ELISA for anti-dsDNA was higher than that of IIF. By comparison with IIF the positive percentages in SLE, MCTD and RA groups were 23 4% vs 17 2%, 12 5% vs 12 5% and 11 8% vs 5 9%, respectively, and the coincident rates between the 2 methods were 93 8%, 100% and 94 1% respectively. The sensitivity and specificity of the developed ELISA for detection of anti-dsDNA were 100% and 93 4% when IIF was as gold standard.Conclusion The ELISA by using plasmid DNA as antigen to detect anti-dsDNA has fine precision, sensitivity and specificity. Its positive rate is higher than that of IIF thus it will contribute to monitor the activities for SLE patients′ condition.

BACKGROUND: Now, dual-energy X-ray absorptiometry is international y recognized as gold standard for the diagnosis of osteoporosis, but the errors can be found in the measurement results due to the heterotopic ossification and bone hyperplasia exists in the measurement part. OBJECTIVE: To investigate the clinical significance of bone metabolic markers in the diagnosis and treatment of elderly patients with osteoporotic fractures, and to research its correlation with the changes of pathological histology and bone mineral density. METHODS: Four bone biochemical markers in 50 elderly patients with osteoporosic fractures were measured preoperatively. According to the results, 25 patients had significantly increased tartrate-resistant acid phosphatase 5b (considered as the increased tartrate-resistant acid phosphatase 5b group), and 25 patients had increased bone alkaline phosphatase (considered as the increased bone alkaline phosphatase group). During operation, the bone tissues of eight patients in each group were treated with hematoxylin-eosin staining and electron microscopy scanning in order to detect the pathological changes. After operation, the patients in the increased tartrate-resistant acid phosphatase 5b group received salmon calcitonin anti-osteoporosis treatment, and the patients in the increased bone alkaline phosphatase group received the anti-osteoporosis treatment of bone peptide injection. The bone mineral density and the four bone biochemical markers were detected again at 6 months after treatment. RESULTS AND CONCLUSION: There were no significant differences in the preoperative bone mineral density and four biomechanical markers between two groups (P > 0.05). The pathological examination results of bone tissue on the fracture site showed that the number of osteoblasts was reduced and the number of oeteoclasts was increased in the increased tartrate-resistant acid phosphatase 5b group; while in the increased bone alkaline phosphatase group, the pathological examination results showed the number of osteoblasts was reduced; the trabecular bone/bone area ratio was decreased in two groups, and there was a significant difference in the decrease degree between two groups (P < 0.05). The electron microscope scanning showed that the osteoclasts of two groups were more active than that of the normal group. The sloppy of trabecular bone in the increased tartrate-resistant acid phosphatase 5b group was more obvious than that in the increased bone alkaline phosphatase group, and the absorption vacuoles were increased. There were significant differences in the bone mineral density and four biomechanical markers between two groups before and after anti-osteoporosis treatment (P < 0.05). The detection of bone metabolic markers could help us to make it clearly that the main function of osteoblast reduce or osteoclast increase in bone tissue of patients, and guide us to use anti-osteoporosis drugs in target. Pathological histology examination can better reflect the condition of osteoblasts, osteoclasts and trabecular bone in bone tissue on the fracture site. Target application of anti-osteoporosis drugs in the osteoporosis patients can effectively improve the efficacy and reduce the relative complications.

Objective To study the clinical value of mammography and ultrasound needle localization biopsy technique in clinical nonpalpable breast disease.Methods 148 patients with negative clinical palpation were operated only because of the mammography or ultrasound findings of lesions.They were undergone localized excision biopsy and derived definite pathological diagnosis.By histopathology diagnosis,benign and malignant lesions,lesion size,complete resection rate were statistically analyzed.Results 21 cases in 148 cases were malignant lesions(14.2％),which were all early breast cancer.The other 127 cases were benign breast disease(85.8％).147 cases were successfully resected at the first time and 1 case was successfully resected at the second time.Conclusion Mammography and Ultrasound fine needle localized biopsy in nonpalpable breast disease diagnosis and treatment has great significance,which can raise early diagnosis rate of breast cancer without mass,and this method is simple and easy for promotion.

Objective To summarize and analyze the adverse reactions caused by cancer chemo-therapy drugs in clinical practice and the specific influencing factors,so as to make more scientific and ef-fective chemotherapy for the treatment of cancer patients.Methods 80 cases of cancer patients treated in our hospital from December to October 2013 in were selected as the research objects.Results Adverse effects of cancer chemotherapy drugs emerged strictly speaking is inevitable,the patient should be the high-est incidence of bad hair in the digestive system,boil down to:nausea,loss of appetite,alopecia and diar-rhea.Conclusions Fully grasp the application of chemotherapeutic drugs adverse reaction and its influen-cing factors,and for making treatment scheme,for improving the treatment effect has important clinical sig-nificance.

Objective To detect the physico-chemical properties , cytotoxicity and antimicrobial activity of dressings containing Ag +.Methods The morphology of this dressing was shown by the scanning electron microscope ( SEM ) .The cytotoxicity was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) method.Using the shake flask test method , the antibacterial effect of dressings was studied .Result There was no significant difference in the water vapor transmission rate between the dressings (P>0.05).Compared with the alginate calcium dressing ,silver ion dressings had a stronger swelling rate .A higher ion concentration would lead to a much larger swelling rate and slower degradation . A lower cytotoxicity was exhibited among the dressings .The silver ion dressing had stronger bacteriostasis to Gram-positive ( G+) and Gram-negative ( G-) bacteria than the alginate calcium dressing .Conclusion The experiment has proved that the silver ion dressing has stronger antibacterial activity and lower cytotoxicity , and it is more effective for wound surface healing, with a shortened treatment course .

Objective To determine the plasma concentration of cytarabine(Ara-C) in children with leukemia and obtain dynamics parameters, and investigate the relationship between the parameters and clinical effect in order to provide the basis for optimization of Ara-C application. Methods Using highperformance liquid chromatogram (HPLC) to determine the plasma concentration of Ara-C, its metabolite Ara-U and infusion rate in 37 children with acute leukemia, their therapeutic reaction, remission, treatment-related infection, side-effect and long-term treatment effect were analyzed in statistic. Results Ara-C by 1～2 g/m2 intravenous drop infusion for 2 hours, the peak plasma concentration time was 2 h and peak concentration were (14.37-84.44)μmol/L, and the median was (41.42±22.80)μmol/L. The median infusion rate was 869.57at 30 minutes after Ara-C drip completion, its average level was (253.40±81.49) μmol/L, over six-times than Ara-C peak concentration. The median continuous complete remission time in 37 children was 29.8 months (5.0～53.1 months), 3y-DFS was (90.63±5.15)%. The therapy-related infection rate was 56.8 %(21/37),including three children (8.1 %) suffered from severe infection, but there was no therapy-related death and no children were off the protocol due to poor tolerance. Conclusion As post-remission treatment, high-dose Ara-C would not cause cumulation in vivo in children with acute leukemia and side-effect were slight. Ara-C could improve the long-term continuous complete remission rate and clinical cure rate for children with leukemia. Therefore, it was worth to apply in clinical.

Objective To detect serum protein spectrum in gastric cancer, screen the serum proteins related to gastric carcinoma and build auxiliary diagnosis model with SELDI-TOF-MS, Methods The serum proteomic fingerprints were detected by SELDI-TOF-MS and CM10 proteinchip. The sera came from 50 first visit gastric cancer patients who were diagnosed by pathological method and 16 healthy peo- ple. The data were analyzed by Biomarker Wizard Software and the proteomic fingerprint of gastric carcinoma model was developed by BPS (biomarker pattern software). Results Compared with the healthy people group, there were 34 significant different protein peaks in the gas- tric carcinoma group. And the diagnosis model composed with 6 proteins (M/Z values were 6016, 6744, 2822, 4474, 7892 and 3242) could classify the 2 groups correctly. In the test group, the sensitivity and specificity were 96% and 93.75% respectively, the accuracy was 95.45%. Conclusion SELDI-TOF-MS method show features such as microcontent, fast and high- resolution etc. It could be utilized to screen significant proteins and develop an auxiliary diagnosis model in gastric carcinoma, which may be used to diagnose this disease.

Objective to investigate the level of platelet leukocyte aggregates in patients with acute cerebral infarction and their short term prognosis.Methods 105 patients with acute cerebral infarction onset within 24 hours were selected continuously,then platelet leukocyte aggregates including neutrophil aggregates (PNA) and platelet monocyte aggregates (PMA) and platelet lymphocyte aggregates (PlyA) were detected by flow cytometry within 24 hours of admission and the incidence of 14 days.modified Rankin Scale(mRS) was performed at 14 days of onset,as a prognostic indicator,and the mRS score was good at 3.The score >3 mRS was divided into poor prognosis.The level of platelet leukocyte aggregates was detected in 50 healthy subjects.Results (1) The platelet leukocyte aggregates in patients with acute cerebral infarction group were significantly higher than that of the healthy group,which was statistically significant (P<0.05).(2)MRs score <3 group and mRS score >3 score comparison,the difference of white blood cell aggregates was statistically significant(P<0.05).Conclusion leukocyte aggregates could be used as an index of short-term prognosis in patients with acute cerebral infarction.

Objective To investigate the clinical characters and therapy of primitive neuroectodermal tumor(PNET).Methods A retrospective analysis was conducted.A 36-year-old female patient was showed pain and numbness of the right upper limb and back for 6 months.The cervical spine MRI showed a spindleshaped intradural mass right ventrolateral of spinal cord at C5-7 with in homogeneously enhancing.Surgery and pathologic examination confirmed that was PNET.Combiled with a series of literatures to analyse the clinical characters Results Surgery was performed to remove the tumor and decompression combined with radiotherapy.The pathologic examination and immunohistochemical analysis revealed that it was PNET.MRI identified local recurrence in spinal canal at 3 month later after surgery.Conclusion Spinal PNET is an uncommon intraspinal tumor with poor prognosis.Histopathology is the evidence of diagnosis.Optimal therapy has not yet been found.Surgical resection with the combination of chemo-radiotherapy or radiotherapy might get the better outcomes.Multidisciplinary treatment should be further clinical required.