The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.

Electromagnetic Fields ; Magnetic Fields ; Microwaves ; adverse effects ; Television

OBJECTIVE To study the isolation and resistance tendency of Acinetobacter calcoaceticus-baumannii to antimicrobial agents from 1998 to 2004 to provide valuable data for infection prevention and therapy. METHODS We reviewed the isolation rates,distribution in clinical specimens and wards,and the resistance rates of(A.calcoaceticus-baumannii)to 14 kinds of antimicrobial agents from 1998 to 2004. RESULTS There was an increasing tendency of isolation rates of A.calcoaceticus-baumannii every year,which was 0.18% in 1998 but 1.48% in 2004.In the seven years,there was the highest isolation rate of 70.58% in specimens from respiratory tract,the next was from the urine(9.42%),and blood(4.63%).Concerning the wards distribution,ICU had the highest rate of 47.28%.In 1998,A.calcoaceticus-baumannii had resistance rates more than 50% only to one kind of antimicrobial agents(aztreonam),but in 2004,it had increased to thirteen kinds(except cefoperazone/sulbactam).About the fourteen kinds of antimicrobial agents we inspected,that were increased in their resistance rate.The highest increasing of resistance rate was ceftazidime from 11.1% in 1998 to 88.9% in 2004,the imipenem was second for 0.0% to 64.8%,and the third was sulfamethoxazole/trimethoprim form 0.0% to 64.0%,while there still was an increasing resistance tendency to them. CONCLUSIONS The clinical isolation rate of A.calcoaceticus-baumannii is increasing,and it has higher resistance rates to many antimicrobial agents as well as an increasing resistance tendency to relatively susceptive antimicrobial agents every year.So physicians should prescribe on the basis of antimicrobial agents susceptibility tests in vitro.

Objective To explore the changes of S - 100? protein in cerebrospinal fluid and serum of children with viral encephalitis and its clinical significance. Methods The levels of S - 100? protein of cerebrospinal fluid and serum of 36 children with viral encephalitis and 20 lumbar anesthesia children without central nervous system diseases were measured by enzyme - linked immunosor bent assay. Differences in the levels of cerebrospinal fluid and serum S-100? protein between children with and without coma, with and without convulsion, with and without sequelae in the case group were compared. Results S-100? protein levels of cerebrospinal fluid in the case group and control group were (0.641?0.390) and (0.037 ? 0.014) ?g/L( P

OBJECTIVETo demonstrate the impact of hypoxia on ER-alpha in both breast cancer tissue and cell line, and its relationship with hypoxia-related parameters.

METHODSExpression of ER-alpha in 51 breast cancer patients with ER positive determined by ligand-binding assay was examined by immunohistochemistry and compared with CA-IX and Glut-1. Impact of hypoxia on breast cancer cell line MCF-7 (ER-alpha positive) was observed by Western Blot and RT-PCR.

RESULTSOf 51 breast cancer patients, 49 were ER-alpha positive. Regional decrease of ER-alpha expression was consistently observed in peri-necrotic regions as compared to distant regions in both in-situ carcinomas (n=29, P <0.0001) and invasive carcinomas (n=20, P=0.0001), which was closely associated with the induction of CA-IX and Glut-1 in hypoxia (P <0.0001). The decreased expression of ER-alpha protein and mRNA in breast cancer cell lines were attributed to hypoxia and not to other stress factors, such as reduced glucose, low pH, and products released from necrotic or hypoxic cells. Chronic intermittent hypoxia could cause persistent down-regulation of ER-alpha in the MCF-7 breast cancer cell line.

CONCLUSIONRegional hypoxia in breast cancer is associated with the reduced ER-alpha expression, and intermittent hypoxia can cause persistent down-regulation. Hypoxia may therefore contribute to the progression of ER-alpha negative status and potentially to the development of resistance to endocrine therapy.

Antigens, Neoplasm ; metabolism ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carbonic Anhydrase IX ; Carbonic Anhydrases ; metabolism ; Carcinoma in Situ ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Cell Hypoxia ; Cell Line, Tumor ; Down-Regulation ; Estrogen Receptor alpha ; genetics ; metabolism ; Female ; Glucose Transporter Type 1 ; Humans ; Hypoxia ; metabolism ; Monosaccharide Transport Proteins ; metabolism ; RNA, Messenger ; biosynthesis ; genetics

Objective To investigate a shorter and safer route for contralateral C_7 nerve root transfer. Methods Eight male patients were treated from Dec2005 to Nov.2006Their range of age was from 22 to 43 years with an average of 30 yearsFive cases had total brachial plexus avulsion.The operative delay was from 2 to 6 months(mean:4 months).The sealenus anterior muscle was transected before a prespinaJ & ret- ropharyngeal tunnel was madeThe contralateral C_7 nerve root was used to repair the upper trunk or the infra- clavicular lateral cord and posterior cord of injured side via this routeusing direct anastomosis or nerve graft- ing.Results The length of the harvested contralateral C_7 nerve root was(4.67?0.52em in the early 5 casesThe nerve graft was6.25?0.35)em long for repairing supraclavicular brachial plexus and(8.56?0.45cm long for repairing infraclavicular brachial plexusThe length of the harvested contralateral C_7 nerve root averaged 6.85cm in the other 3 cases2 of which had direct anastomosis to the residual nerve C_5 and C_6 nerve roots and the other used nerve graft of 3 cm in lengthTransient contralateral sensory symptoms were re- ported in most patientsAt 3 months followups6 patients had tingling sensation on the contralateral fingers with percussion on the injured cervical areaIpsilateral SSEP could be recorded by stimulating at 2 cm above sternoclavicular joint on the injured sideAt 7 months follow ups of 5 patientsCMAP could be recorded in bi- cepsdeltoids and infraspinatus or triceps with stimulation at Erb's pointHoweverno clinical movements was noticedAt 12 months follow ups of 3 patientswe could observe early motor and sensory function recovery of those patients to different extent.Conclusion Transection of anterior scalenus muscle shortens the length of the transfer route and allows more efficient neurotizationThe procedure is convenient and safeprovided certain precautions being usedThe principal of contralateral C_7 nerve transfer are reconstruct the anterior divi- sions of upper trunkposterior divisions of upper trunk and suprascapular nerve when repairing the supraclavic- ula brachial plexusReconstruct the lateral cord and posterior cord when repairing the infraclavicula brachial plexusPostsurgical fasting for 4 days included foods and liquids will benefit of healing of anastomosed nerves and regenerationaud avoid complications.

Objective To investigate the cerebral blood flow (CBF) perfusion patterns and related factors in hyperthyroidism patients.Methods Twenty-five patients with hyperthyroidism and twenty-two healthy controls matched for age,sex,education were enrolled.~(99)Tc~m-ethylene cysteinate dimer (ECD) SPECT CBF perfusion imaging was performed at rest.Statistical parametric mapping 5.0 software (SPM5) was used and a statistical threshold of P＜0.05 (corrected) was applied for signifying changes of regional CBF (rCBF).The semiquantitative values of rCBF were extracted automatically by brain search 1.1 software and were correlated with concentrations of serum thyroid hormones(FT_3,FT_4),thyroid autoimmune antibodies:sensitive thyroid stimulating hormone(sTSH),thyroid peroxidase antibody (TPOAb) and TSH receptor antibody (TRAb) by Pearson analysis,with disease duration by Spearman analysis.Results rCBF was decreased significantly in limbic system and frontal lobe,including parahippocampal gyrus,uncus (posterior entorhinal cortex,posterior parolfactory cortex,parahippocampal cortex,anterior cingulate,right inferior temporal gyrus),left hypothalamus and caudate nucleus (P＜0.05,corrected).rCBF in left lingual gyrus,posterior cingulated was negatively correlated with concentration of FT_3(r=-0.468,-0.417,both P＜0.05).rCBF in left lingual gyrus,bilateral inferior temporal gyrus,right superior parietal lobe was negatively correlated with concentration of FT_4(r=-0.4M,-0.418,-0.415,-0.459,all P＜0.05),while that in left mammillary body and putamen was positively correlated with concentration of FT_4(r=0.419,0.412,both P＜0.05).rCBF in left insula was negatively correlated with concentration of sTSH,and right auditory associated cortex was positively correlated with concentration of sTSH(r=-0.504,0.429,both P＜0.05).rCBF in left middle temporal gyrus,left angular gyrus was positively correlated with concentration of TRAb while that in right thalamus,right hypothalamus,left anterior nucleus,left ventralis nucleus was negatively correlated with concentration of TRAb(r=0.750,0.862,-0.691,-0.835,-0.713,-0.759,all P＜0.05).rCBF in right anterior cingulate,right cuneus,right rectus gyrus,right superior marginal gyrus was positively correlated with concentration of TPOAb(r=0.696,0.581,0.779,0.683,all P＜0.05).rCBF in postcentral gyrus,temporal gyrus,left superior marginal gyrus and auditory associated cortex was positively correlated with disease duration(r=0.502,0.457,0.524,0.440,all P＜0.05).Conclusion Hypoperfusions in limbic system and fontal lobe were found in hyperthyroidism Patients,which might be associated with thyroid function and disesse duration.

·AIM: To observe the therapeutic effect of visual perception training combined with total nutrition meal sequential therapy in the treatment of myopic amblyopia.·METHODS:Totally 73 children ( 135 eyes ) with myopic amblyopia were divided into control group ( 36 cases, 67 eyes) and treatment group (37 cases, 68 eyes) according to random number table. The control group were treated with traditional spectaculars and grating covering combined with fine eyesight training;the treatment group were treated with visual perception training combined with total nutrient meal sequential therapy. The visual acuity, diopter and average diopter of two groups were compared before and after treatment at 3, 6mo and 1a. The curative effect of two groups of children was compared after 1a treatment. And the adverse reactions were recorded in two groups during the treatment period. The recurrence rate of amblyopia in 1a follow-up was compared between two groups.·RESULTS: The difference of visual acuity between two groups was not significant at 3mo (P>0. 05). The visual acuity of the treatment group was significantly higher than that of the control group at 6mo and 1a (P<0. 05). There was no significant difference in diopter between the two groups after 3, 6mo and 1a (P>0. 05), but the average annual refractive changes in the treatment group were significantly lower than that in the control group ( P<0. 05). The basic cure rate and total effective rate of the treatment group were significantly higher than that of the control group (P<0. 05). There were no severe adverse reactions occurred between two groups during the treatment period. The recurrence rate of amblyopia in the treatment group was significantly lower than that in the control group (P<0. 05) after 1a follow-up.· CONCLUSION: Visual perception training combined with total nutrition meal sequential therapy in the treatment of myopic amblyopia in preschool children can significantly improve patients' visual acuity, reduce the average annual diopter changes, improve the therapeutic effect, reduce the recurrence rate of amblyopia.

Objective To investigate the added diagnostic value of SPECT/CT imaging over routine planar whole-body bone scintigraphy (WBBS) for bone metastases from lung cancer. Methods One hundred and forty-six patients with lung cancer, confirmed by pathological examination, underwent routine 99Tcm-MDP (1110 MBq) WBBS, followed by SPECT/CT over the regions with indeterminate findings on WBBS. Both WBBS and bone SPECT/CT images were interpreted by two experienced nuclear medicine physicians in consensus as the positive, negative or uncertain bone metastases. The final diagnosis was comfirmed by pathology or clinical follow-up. x2 test was used to compare the differences between the two imaging methods. Results Finally, 45 patients were diagnosed as positive bone metastases and the other 101 as negative. The diagnostic sensitivity of bone SPECT/CT for bone metastases from lung cancer was 93.3%(42/45), singnificantly higher than that of WBBS (64.4%, 29/45) (x2 = 19.944, P＜0.05). The diagnostic accuracy of bone SPECT/CT was 89.7% ( 131/146), much higher than that of WBBS (44.5%,65/146) ( x2 = 69. 598,P ＜ 0.05). The uncertain and incorrect diagnostic rates of bone SPECT/CT and WBBS were 10.3% ( 15/146, raging from 5.3% to 15.2% with 95% confidence interval (CI) ) and 55.5% (81/146, raging from 47. 4% to 63.5% with95% CI), respectively. Conclusion BoneSPECT/CT provides incremental diagnostic value over routine WBBS for bone metastases from lung cancer.

Objective To investigate the additional diagnostic value of 99Tcm-MDP SPECT/CT over conventional SPECT scan in patients with spinal bone malignancy. Methods Fifty-two patients (mean age (56±14) years) with suspicious spinal bone diseases underwent both bone SPECT and SPECT/CT imaging right after 99Tcm-MDP whole-body planar bone scintigraphy. All patients were pathologically diagnosed by either spine operation or trans-spinal biopsy. The images were evaluated by two independent reviewers; inter-reviewer agreement was evaluated using a weighted Kappa score. Each focus of abnormality was recorded using a 4-point diagnostic confidence scale: benign (B), possibly benign (PB), possibly malignant (PM) or malignant (M). Results Accord to the pathological results, 36 patients had bone malignancy (19 with metastatic tumors and 17 with non-metastatic tumors) and 16 had benign lesions. Over SPECT images, two reviewers rated 73.1% (38/52) and 67.3% (35/52) the lesions as the equivocal (PB or PM) respectively. Over SPECT/CT images, they rated only 25.0% (13/52) and 13.5% (7/52) lesions as the equivocal. Inter-reviewer agreement was 63.5% (weighted Kappa score=0.62) for SPECT and 78.9% (weighted Kappa score=0.81) for SPECT/CT. Conclusions Compared with99Tcm-MDP SPECT, 99Tcm-MDP SPECT/CT results in a significant reduction of equivocal diagnoses for the spinal bone malignancy.