A study to construct three prokaryotic expression vectors and its expression in adenosine deaminase / 国际检验医学杂志

Objective To express adenosine deaminase protein by molecular cloning technique.MethodsTotal RNA was extracted from human leukocytes and the cDNA was obtained by reverse transcription.Whereupon the cDNA was used as a template to amplify adenosine deaminase gene by polymerase chain reaction (PCR) and then integrated it into three prokaryotic plasmids pET-28 b, pET-32 a (+) and pHSIE.The plasmid with the correct sequencing was transformed into E.coli BL21 (DE3) by CaCl2 method for the protein expression.The expression activity of these fusion proteins were detected by Western-blot and SDS-PAGE, with the optimized expression conditions.Results Complete fusion of target gene and three prokaryotic plasmids was observed through sequencing.The expressed and accurate ADA protein was identified by Westernblot and SDS-PAGE.The optimal expression conditions were observed:the protein expression would be induced with 0.4 mmol/L IPTG and incubated at 16℃for 24 hours.Conclusion The prokaryotic vectors of adenosine deaminase (BL21+pET-28 b+ADA, BL21+pET-32 a+ADA, BL21+pHSIE+ADA) were successfully constructed and efficiently expressed.

Detection of TORCH infection in pregnant women by using reverse phase protein array / 国际检验医学杂志

Objective To evaluate whether the reverse phase protein array (RPPA) method can be used for detecting TORCH infection in pregnant women .Methods The RPPA method was established for detecting TORCH infection .The positive coinci‐dence rates of TORCH infection detected by the RPPA method and ELISA method in 2000 fresh serum samples from pregnant women were compared for evaluating the feasibility of RPPA in TORCH detection .Results The positive coincidence rates of estab‐lished RPPA and ELISA for detecting TORCH infection was 100 .0% ,91 .1% ,97 .2% ,91 .3% and 93 .0% respectively ,indicating that the detection results of various indexes by RPPA and ELISA had better consistency (P>0 .05) ,but the positive detection rates of RPPA for Rubellavirus ,CMV and HSV‐1 ,2 were higher than those of correspondent ELISA method .Conclusion RPPA method for detecting TORCH infection has the advantages of simpleness ,rapidness ,high sensitivity and strong specificity ,is an effective method of auxiliary diagnosis for bearing and rearing better children in clinical ,and is worthy of being promoted and used in the fu‐ture .

Application of Sysmex XE-5000 automated hematology analyzer in cell counting of serous cavity effusion / 国际检验医学杂志

Objective To investigate the application of hematology analyzer in cell counting of serous cavity effusion .Methods Humoral mode of Sysmex XE-5000 automated hematology analyzer and manual microscopy were employed to perform cell counting in 85 samples of serous cavity effusion .Results Compared with the manual method and instrument method ,differences of mononu-clear cells and multinucleated cells of serous cavity effusion showed no statistical significance (P>0 .05) ,while those of leukocytes , erythrocytes were found statistical significance (P<0 .05) .Conclusion Sysmex XE-5000 automated hematology analyzer has advan-tages of simple ,stability and accuracy ,however ,its application in the effusion cell counting can not completely replace the manual microscopy .

The role of IDO/TTS mediated tryptophan metabolic pathway in patients with immune thrombocytopenia / 实用医学杂志

Objective To discuss the role of indoleamine 2, 3-dioxygenas e (IDO) and tryptophanyl-tRNA synthetase (TTS) mediated tryptophan catabolism in immune thrombocytopenia (ITP) patients treated with high doses of dexamethasone through the expressions of IDO and TTS in T cells , and the concentrations of plasma kynurenine and tryptophan. Methods 20 newly diagnosed or relapse ITP patients were treated with 40 mg/d × 4 d dose of dexamethasone. The heparin anticoagulant blood samples were obtained before treatment and the 5th day after treatment. 20 healthy subjects were selected as the control group. The IDO and TTS expressions in CD4+and CD8+ T cells were analyzed by flow cytometry. The concentrations of plasma kynurenine and tryptophan were detected by liquid-mass spectrometry system. Results Compared with healthy controls group, the plasma tryptophan and kynurenine concentration and the ratio of Kyn/Trp were significantly elevated in ITP patients (P <0.05); the IDO expressions of CD4+ and CD8+ T cells in ITP patients were significantly lower than those in healthy controls (P < 0.05), but the TTS expressions were significantly higher (P < 0.05). The concentration of tryptophan in effective group was significantly lower than before treatment (P < 0.05), in contrast, the kynurenine concentration and the ratio of Kyn/Trp were significantly higher than before (P < 0.05). The expression of IDO in effective group were significantly higher than that before treatment (P < 0.05), conversely, the expression of TTS in effective group were significantly decreased (P < 0.05). No significant difference can be found in ineffective group. Conclusion IDO/TTS-mediated tryptophan catabolism pathway could indicate the onset of ITP. The sensitivity of ITP patients with high dose of dexamethasone treatment can be observed through the level of IDO and TTS.

Nano-resin and self-etching adhesive for bonding repair of anterior tooth defects with vital pulp:A 2-year follow-up / 中国组织工程研究

BACKGROUND:Compared with the conventional composite resin, the 3M Z350 nano-resin has good wear resistance, physical mechanical properties, and polishing, and exerts a lower irritation to the dental pulp. Besides fil ing materials, a reliable tooth-prosthesis bonding interface is necessary for resin bonded repairs. OBJECTIVE:To compare the clinical effects of self-etch bonding Adper Easy One and total-etch bonding Single Bond 2 on nano-resin bonding restoration of the anterior teeth. METHODS:120 anterior teeth with vital pulp, which had defects at the incisal ends and were to be restored with nano resins, were divided into two groups randomly. Two kinds of adhesives, self-etch adhesive and total-etch adhesive, combined with nano-resin were used to restore the teeth. The patients were re-examined immediately, 6 months, 1 year and 2 years after the treatment. The fil ings, teeth and pulps of patients were examined, including whether the prosthesis and tooth color were coordinated, whether the gap between the prosthesis and the teeth were sealed, whether the surface of the prosthesis was intact with no loose, whether the prosthesis and teeth had no staining and secondary caries, whether the condition of the tooth pulp had hot or cold stimulation-induced pain. RESULTS AND CONCLUSION:No significant difference in the fil ing effects was found between the two groups when the patients were re-examined immediately, 6 months and 1 year after the treatment (P>0.05). The pulp lesions of the self-etching group were fewer than those of the total-etch group 2 years after the treatment (Pincomplete restoration and secondary caries, respectively. No statistical y significant differences were found in these four aspects between the two groups (P>0.05). The 2-year fol ow up showed a low incidence of pulp lesions and satisfactory clinical performance after 3M Z350 nano-resin working with self-etching bonding system in the nano-resin fil ing of anterior teeth with vital pulp.

Detection of serum MDA, NO and iNOS in patients with SARS and its clinical significance~* / 中国病理生理杂志

AIM: To observe free radicals (MDA, NO) and iNOS of patients with severe acute respiratory syndrome (SARS) and to explore its significance. METHODS: MDA, NO_2~-/NO_3~- and iNOS were determined in SARS patients during the early, recovery and follow-up stage, front doctors and nurses (contact group) and health people (health control). RESULTS: The level of MDA during first stage was higher than that of recovery stage and the MDA level of recovery stage was higher than that of follow-up stage, contact group, and health control group (P0.05). CONCLUSION: The pathological injury in pathogenesis of SARS is related to free radicals. [

Differential expressions of ameloblastoma related genes determined by microarray / 中国病理生理杂志

AIM:This study aims to screen the differentially expressed genes related to the pathogenesis of ameloblastoma by cDNA microarray.METHODS:The total RNAs were isolated from ameloblastoma and tooth germ,respectively.The RNAs were purified by oligotex.Both the mRNAs from two kinds of tissues were reversely transcribed to cDNA with the incorporation of fluorescent-labelled dUTP to prepare the hybridization probes.The mixed probes were hybridized to cDNA microarray.Tumor-related genes were screened through the analysis of fluroescent intensity.RESULTS:722 genes exhibited significant changes in expression levels in the ameloblastoma in comparison with tooth germs tissues.240 genes were overexpressed more than doubled(92 genes were more than 3-fold),and 482 genes were underexpressed to below 0.5 of the control level.CONCLUSION:Microarray technique facilitates large scale and rapid identification of potential target genes of ameloblastoma.

Detection of serum IFN-?, IFN-?, NO and iNOS in patients with SARS and its clinical significance / 中国病理生理杂志

AIM: To study the serum level of interferon ( IFN)-?, IFN-?, nitric oxide (NO) and nitric oxied synthase (iNOS) in severe ac ute respiratory syndrome (SARS) patients and to expl ore its significance. METHODS: IFN-?, IFN-?, NO and iNOS were determined in SARS patients during the early, recovery and follow-up stage, the health doctors and nurses and health people, respectively. RESULTS : The serum level of IFN-? during the early stage was higher than that of other groups (P