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Mitochondria play a central role in the regulation of energy metabolism and signal transduction in eukaryotic cells. Although many fluorescent labeling strategies have been developed for mitochondrial studies, the methods that enable labeling efficiency assessment at the single-mitochondrion level are still lacking. By employing the unique advantages of high sensitivity flow cytometry ( HSFCM ) in the sensitive, rapid, and quantitative multiparameter analysis of individual mitochondria, here we examined the performance of several different mitochondrial labeling strategies from the perspectives of brightness, labeling ratio, and stability. Mitochondria isolated from HeLa cells transfected with pAcGFP1-Mito plasmid upon transient or stable transfections, and mitochondria directly labeled with MitoTracker Green or SYTO 62 were analyzed by a laboratory-built three-channel HSFCM. Upon the quantitative measurement of fluorescence brightness, it was found that the fluorescence intensity of green fluorescent protein ( GFP ) in mitochondria isolated from cells with stable transfection was about 17. 7-fold higher than the transient transfection ones, and was approximately two orders of magnitude brighter than mitochondria labeled with MitoTracker Green. On the other hand, the fluorescence signal of SYTO 62 labeling decreased upon washing, indicating its rapid dissociation rate. The strong fluorescence intensity and good labeling stability make stable transfection an efficient method to label mitochondria. The experimental results demonstrates that HSFCM provides a powerful analytical tool to assess the performance of mitochondrial fluorescence labeling via high throughput single mitochondria analysis.
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Objective To investigate the antimicrobial susceptibility and molecular nature related to the resistance on macrolides from nasal Staphylococcus (S.) aureus isolates among healthy people.Methods A total of 100 S.aureus isolates collected from 2009 to 2011 were tested for antimicrobial susceptibility by E-test.Double disc test (D-test) was used to detect the inducible clindamycin resistance.All S.aureus isolates were characterized by spa typing.Macrolides resistance genes were detected and compared with isolates that were collected clinically or from the livestock.Results High resistance rates on erythromycin or clindamycin was noticed,with 52％ and 27％,respectively.Inducible clindamycin resistance was identified in 29 of the 100 (29％) isolates.In total,the 100 isolates were assigned to 35 spa types.The most common spa types were found to be t189,t571,t002,t796,t437,t034 and t701,that accounted for 51.0％ of all the isolates.erm (C) (57.7％) and erm (B) (34.6％) were found as the dominant genes in 52 S.aureus isolates from healthy people.On the other hand,erm (A) and erm (C) were identified in 95.0％ S.aureus isolates from patients and all the livestock,respectively.Conclusion erm (C) and erm (B) carrying S.aureus strains were circulating in healthy people and these genes were distributed in different S.aureus clones.
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Mitochondria play a central role in the regulation of cellular energy metabolism, bio-synthesis and cell death. Dysfunction of mitochondria can lead to many diseases. Mitochondrial proteomics provides important theoretical foundation for a systematic understanding of the biological functions of mitochondria, studying the mechanisms of mitochondria-related diseases, and promoting the research and development of mitochondria-targeting drugs. The methodologies, recent technology development, and characteristics and applications of mitochondrial proteomics were reviewed and the challenges and prospects were also discussed.
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Objective:According to the tumor of children psychology, explore a set of suitable for children accurate tumor radiotherapy positioning method, in order to improve the positioning accuracy, thus improve the tumor clinical cure rate.Methods: 1.Eliminate fear; 2.Adaptability training; 3.The methods of psychological suggestion. Evaluation method: take psychological induction of radiation induced by radiation therapy in children with and without taking mental children from the placement time, downtime in drug utilization rate, treatment rate and bit error rate comparison.Results: There is obvious difference between two groups (P<0.05).Conclusion: radiotherapy positioning of children with psychological intervention in the method, can improve the treatment of children's compliance.
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Objective To investigate the clinical effects of comprehensive rehabilitation treatment for patients with a diabetic foot. Methods Fifty-two patients with one diabetic foot were randomly divided into a rehabilitation group ( n =26 ) and a control group ( n =26).The patients in the control group received conventional treatment including health education,basic medical therapy and focal treatment of the foot.In addition to the conventional treatment,the rehabilitation group was treated with aerobic exercise,Beecher's exercise regimen,infrared therapy and ultrashortwave therapy 6 days a week for 4 weeks.The therapeutic effect and quality of life were evaluated before and after treatment. Results The effectiveness rate of the rehabilitation group was significantly higher than that of the control group ( 88.46％ vs 73.08％ ) after treatment.The quality of life scores in both groups were significantly better than those before treatment,including on the social relationship dimension,the physiological function dimension,the mental/psychology dimension and the therapy influence section.Average quality of life scores in the rehabilitation group had improved significantly more than in the control group. Conclusion Comprehensive rehabilitation is beneficial to prevent the progress of diabetic foot and to improve the quality of life of patients with a diabetic foot.
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Objective To analyse the survival time and related factors of patients with brain stem glioma who received 3DCRT.Methods Thirty-six patients with brain stem tumor were admitted from October 2004 to December 2008 and all received 3D-CRT with the dosage (50-54 Gy,25-30 f,5-6 weeks).During treatment,the patients’ outcomes were analyzed by observing the changes of symptoms,signs and adverse radiotherapy reaction and all of them were followed-up in the next 3 years.The survival data were analyzed by Kaplan-Meire method.Results The median survival time was 9 months in the 23 pediatric patients and 15 months in 13 adult patients.One-,two-and three-year survival rates between pediatric group and the adult group were 43.5 ％ (10/13) vs 76.9 ％ (10/13),26.1％ (6/23) vs 46.2 ％ (6/13),8.7 ％ (2/23) vs 38.5 ％ (5/13).Karnofsky performance scale score at admission (x2 =20.059,P =0.000),tumor site (x2 =17.585,P =0.000),growth pattern (x2 =21.247,P =0.000) were associate with survival time.Conclusion 3DCRT is an effective therapy to brain stem glioma,childhood onset,pontine glioma,diffusion style and Karnofsky performance scale less than 80 are risk factors of poor prognosis.
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Objective To study the expressions of miRNAlet7 and Ras in non-small cell lung cancer ( NSCLC),and their correlations with clinicopathological features and survival time.Methods In-situ hybridization was used to detect the expression of let7,and SP immunohistochemistry to measure HMGA2 in 68 NSCLC cases ( group A) and 20 cases with normal lungs ( group B).Results The positive rate of let7 in group A was lower than that in group B (39.7％ vs 63.2％ ) ( P ＜0.05).The positive rate of Ras in group A was higher than that in group B (66.2％ vs 25.0％ ) ( P ＜0.01 ).The positive rate of let7 was not related to the age,gender,histological type,cell differentiation,and clinical stages of cancer patients(P ＞0.05).The positive rate of Ras was related to smoking,sex,and histological type of cancer( P ＜0.01 ),and was not related to cell differentiation,lymphatic metastasis,and clinical stages of cancer ( P ＞0.05).There was an obvious negative correlation between let7 and Ras( r =-0.627,P ＜0.01 ).The 2-year survival rate of let7-positive group was higher than that of the let 7-negative group ( x2 =4.84,P ＜0.05).No statistically significant difference was found between Ras-positive and-negative groups ( P ＞0.05 ).Conclusions The lower level of let7 expression,and high level of Ras expression have much to do with the carcinogenesis of NSCLC.The level of let7-positive expression is closely related to prognosis; while Ras may act in cooperativity in the occurrence and development of NSCLC.
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@#Objective To investigate the factors that influence the patients to understand the stroke they suffering from.Methods 61 stroke patients were investigated with a questionnaire designed by ourselves.ResultsThe age,sex and educated level affected the stroke patients' understanding,but the time after onset,the degree of locomotory injured and hemiplegic side did not.ConclusionDistinguishing direction of rehabilitation should be established according to the age,sex and educated level of patients.
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@#ObjectiveTo investigation the requirement of patients for occupational therapy, and find a better manner for occupational therapy practices.MethodsThe questionnaire survey of expectation for occupational therapy was performed for sixty-nine patients. Questions were as below: how much does each patient expect through their occupational therapy for the goal, such as physical function recovery, psychological stability, vocational reinstate and so on.ResultsMore than ninety percent subject responded to questions about physical function recovery, re-acquiring roles in own family, decreasing care volume, psychological stability and pastime activity as expected. About the question of condition after discharge, thirty-two percents wanted to live in own home without care; while reinstate, continue to hospitalization, and live in care facilities were twenty-five percents respectively.ConclusionThere is a remarkable higher requirement for physical function recovery, however through the explanation about occupational therapy, another aspects of it are to be recognized.
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@# ObjectiveTo examine the occurrence rate of visuospatial hemi-neglect of patients with cerebral injury with behavioural inattention test (BIT) (Japanese version),and assess the validity of the BIT test.Methods22 subjects (16 with right hemispheric damage and 6 with left hemispheric damage) were tested with BIT. ResultsThe rate of visuospatial hemi-neglect in 22 patients with hemispheric damage was 36.4% (8 cases) , and that rate in 16 patients with right hemispheric damage was 31.3% (5 cases), in 6 patients with left hemispheric damage was 50% (3 cases).ConclusionThe occurrence rate of visuospatial hemi-neglect appears to be independent from male or female, age, course of disease and education level of study subjects.
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Sepharose 6B was activated by epichlorohydrin in the strong base condition, and then reacted with solution of iminodiacetic sodium. The arms of IDA were conjuncted to the activated Sepharose 6B. Then the products were reacted with the solution of NiSO4. The arms of IDA were chelated with Ni2+,and the chelating resin―Ni2+-IDA could be prepared. The physicochemical indexes and performance in purifying protein of the expressing product were assayed with atomic absorption method and purifying aimed protein-human B lymphocyte stimulator(hBLyS) from the expressing products in E.coli. The results indicated that the performance of made gel is very good, and its price is less than 1/10 of that of commodity gel.
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Objective To investigate imaging parameters on low field MRCP and to optimize the quality of images.Methods MRCP in 77 cases were performed using Airis-Ⅱcomfort open 0.3T low field MR imaging system made by HITACHI company.3D-FSE combined with 2D single-shot breath-hold FSE sequences were used.The 3%～5% compound solution of Gd-DTPA was taken to restrain the hige signals from remaining liquid in stomach.Results MRCP examination was succeeded in all 77 cases and the images in all cases but 2 were fine for diagnosis.The total diagnostic accuracy was 79.2%.The accuracy of MRCP in the detection of the degree and location of pancreaticobiliary duct obstruction was 100%. Conclusion The distinct MRCP images can be obtained with low field MR system using combination of several scanning techniques and proper scanning parameters,and also patients have to prepare fully.
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Objective To evaluate the diagnostic value of MRI in hepatic echinococcosis.Methods MRI features of 37 cases with hepatic echinococcosis proved by operation and pathology were analysed in comparison with that of CT.Results The hepatic echinococcosis could be divided into five types according to the manifestations of CT and MRI:simple cyst in 8 cases,multiple daughter cyst in 13 cases,ruptured cyst in 7 cases,consolidated and calcified cyst in 4 cases and mixed cyst in 5 cases.Conclusion MRI can clearly show and determine the location,size,form,number and types of hepatic echinococcosis,that are helpful for the plan of operation.
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Objective To evaluate the diagnostic value of the low field MRCP in obstructive jaundice by comparing its result with that of US and CT. Methods 55 cases of obstruction jaundice patients proved by clinical diagnosis were studied and compared the collected original diagnosis results by low filde MRCP with US and CT.Results Of 55 cases of obstructive jaundice, 30 were cholelithiasis ,21 were tumors and 4 belonged to the other type. The accuracy of MRCP for defining the site of obstruction was 100%. The accuracy for identifying the cause of obstruction was 81.8%.Conclusion The low filde MRCP can accurately define the obstructive site in biliary obstructive disease .When used together with 2D single-shot breath-hold MRCP , T 2WI, T 1WI and 3D-FSE MRCP shows high accuracy and sensitivity in diagnosing obstructive jaundice.