The aim of this study is to clone the mouse T-bet promoter and enhancer, construct and identify the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS for T-bet transcription regulation study and its function in signaling of multiple sclerosis. The promoter and CNS of T-bet were predicted by bioinformatics assay. The predicted fragment of mouse T-bet promoter plus CNS was amplified by PCR and cloned into pGL4.10. The recombinant plasmid pGL4.10-TBX21pr-CNS was transferred into Escherichia coli DH5α. The positive clone was identified by double digestion with Kpn I and Sfi I and DNA sequencing. Finally, pGL4.10-TBX21pr-CNS was cotransfected with pRL-TK into 293T cells and Jurkat cells, pRL-TK and pGL4.10 as a control. The luciferase activity in 293T cells (P = 0.012 2) and Jurkat cells (P = 0.002 2) was higher than that of the control group. A fragment of 1 028 bp mouse T-bet promoter plus 1 308 bp CNS was successfully cloned and the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS was constructed. In 293T cells and Jurkat cells, pGL4.10-TBX21pr-CNS has the promoter functions. This work offers a basic material for the research of T-bet transcription.
Animals ; Gene Expression Regulation ; Genes, Reporter ; Genetic Vectors ; Luciferases ; Mice ; Multiple Sclerosis ; Plasmids ; Promoter Regions, Genetic ; T-Box Domain Proteins ; genetics

Objective To summarize the bacterial drug susceptibility of common enterobacteriaceae in pediatric patients and the epidemic status of the extended spectrum beta‐lactamases(ESBLs) during 2013 and 2014 in our hospital in order to guide the clini‐cal application of antibacterial drugs .Methods Routine methods of isolation and culture were performed and the distribution of en‐terobacteriaceae with resistance were statistically analyzed with the software .Results A total of 530 isolates were collected in the hospital in which was 32 .08% Klebsiella pneumonia and 38 .87% Escherichia coli .The most sensitive drug of Enterobacteriaeeae was carbapenems .ESBLs positive rate in Escherichia coli and Klebsiella pneumonia were 32 .04% and 20% .The difference was sta‐tistically significant (P<0 .05) .Conclusion Escherichia coli and Klebsiella pneumonia were the common enterobacteriaceae in pedi‐atric patients .Carbapenems remained very high activity against Enterobacteriaceae .However ,the carbapenems resistance strains have been appeared that should pay attention .Antibiotic therapy should be based on the guidance of bacteriology to select sensitive drugs .

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)is a powerful tool for routine identification of a variety of microorganisms,including bacteria,yeast and filamentous fungi.Fast,accurate, high-throughput and low-cost characteristics,and gradually used for filamentous fungal infection in the laboratory diagnosis. In the MALDI-TOF MS analysis of filamentous fungi,from sample preparation to obtaining accurate identification results, each step determines whether the medical technician can provide accurate results for the clinician.This review will provide a systematic overview of the applications and prospects of filamentous fungal analysis (including identification,typing,and an-tifungal susceptibility testing)from preparation of the test sample to the MALDI-TOF MS technique,as well as a review of some of its future developments.

Six cases of T-cell lyrnphoma associated with reactive histiocytic hyperplasia with hemophagocytosis (HHH) localized primarily to the skin and subcutaneous fat were described. Initial biopsy findings may he misinterpreted as benign panniculitis. Four of which were progressive, recurrent, even fatal (follow-up for 8-36 months). Histologically the lesions of all cases were composed of a mixture of small and medium sized atypical lymphoid cells and benign hemophagocytes infiltrated with neutrophils and eosinophils between fat cells. The mechanism and differential diagnosis of the disease were discussed.

Wrist-ankle acupuncture is a type of special acupuncture therapy, featured by simple operation, swift efficiency, and few adverse effects. In the recent years, ankle-wrist acupuncture has become more and more popularized in clinic. By retrieving the relevant medical literatures from VIP, CNKI, and Wan Fang databases, this manuscript was to analyze the history of wrist-ankle acupuncture and to review it from both mechanism and clinical reports. Meanwhile, this article also pointed out the existing problems in the research of wrist-ankle acupuncture during the recent years, and expected further standardization and studies.

Objective To compare the results of emergency clinical training for medical students and to put forward some suggestions.Methods Toatlly 207 medical students of Grade 4 studied in the first people' s hospital of Shanghai Jiaotong university from 2009 to 2011 were divided into the traditional group (n =90 ) and the experimental group (n =117 ).The students in the experimental group participated in emergency clinical training in summer vacation while those in the traditional group did not.Questionnaires about their self-learning ability,clinical communicating ability and critical thinking ability after training the were conducted and the data were analyzed by descriptive statistical analysis.Results One hundred and five effective questionnaires were recovered from the experimental group,with the recovery rate 89.7％ and 62 effective questionnaires were recovered from the traditional group,with the recovery rate 68.9％.The scores about clinical communicating ability and critical thinking ability of the experimental group were significantly highcr than those of the traditional group [ (79.45 ± 9.354)vs.(75.87 ± 9.926),(263.38 ± 30.925 ) vs.(251.36 ± 23.679),P ＜ 0.05 ].However,there was no significant difference in scores about self-learning ability between the two groups [ ( 153.97 ± 23.725 ) vs.( 149.83 ± 13.891 ),P ＞ 0.05].Conclusion Emergency clinical training is helpful to improve medical students' clinical communicating ability and critical thinking ability.It is worth popularizing.

The afferent connections of the interpeduncular nucleus (IPN) were studied with retrograde transport method of using horseradish peroxidase (HRP).A microinjected syringe needle or microelectrophoretic pipette was inserted into the brain in the vertical, inclined through cerebellum or inclined through cerebral cortex medially.The results were as follows:Following HRP injection into the IPN, labeled neurons were found in medial habenular nuclei, lateral habenular nuclei, dorsal raphe nucleus, superior central nucleus, dorsal tegmental nucleus of Gudden, dorsolateral tegmental nucleus, ventro-lateral and lateral portions of periaqueductal grey matter, and locus coeruleus. In addition, labeled cells were also observed within caudal portion of accumbens nucleus and nucleus tractus diagonalis (Broca).Following HRP injection into the lateral, ventral (or ventrolateral) and dorsal portions of the IPN, labeled cells were found in ipslateral medial habenular nucleus,medial regions of the bilateral medial habenular nuclei, and lateral regions of the bilateral medial habenular nuclei, respectively.After injection into the rostral or caudal pole of the IPN, no labeled cells were observed in bilateral medial habenular nuclei.Following injection of HRP into the caudal pole of the IPN, labeled neurons were observed within both sides of lateral habehular nucleus, dorsolateral tegmental nucleus and ventromedial portion of dorsal tegmental nucleus.No labeled neurons were observed in the ventral (deep) tegmental nucleus,premamillary nucleus, supramamillary nucleus and parabigeminal nucleus except in the cases in which the HRP injection site extended beyond the IPN.

Afferent connections to the nucleus tegmenti dorsalis were studied by means of retrograde axonal transport method of horseradish peroxidase (HRP). HRP was depositted stereotaxically via three approaches: vertical, oblique through the cerebellum or oblique through the cerebrum. The serial sections were treated by 3,3'-diamino-benzidine, benzidine dihydrochloride, diaminobenzidine-cobalt dichloride or tetrame-thylbenzidine respectively.Labeled neurons were observed in following nuclei: nucleus lateralis habenulae,nucleus interpeduncularis, nucleus mammillaris lateralis, nucleus posterior hypothalami, nucleus lateralis hypothalami, substantia grisea centralis mesencephali, nucleus dorsalis raphes, nucleus tractus solitarii, nucleus prepositus hypoglossi, nucleus parafascicularis and nucleus of Darkschewitsch.Then HRP was introduced into nucleus parafascicularis or nucleus prepositus hypoglossi, labeled terminals were also observed within the nucleus tegmenti dorselis.

Lycobetaine ( AT-1840 ) is an effective antitumor substance. In the present work it was demonstrated that in a dose of 40 pig/kg AT-1840 could decrease the percentage of G, cells, meanwhile it increased G2 + M cells remarkably by means of flow fluocytometry method, AT-1840 could also inhibit the progression of cells from G2 to G1 phase. 8 to 72 h after, the injection the mitotic index was decreased significantly and recovered gradually in 6 d. It could inhibit the prophase and metaphase greatly, but had no marked influence on anaphase & telophase.

Objective: To evaluate the reliability and validity of the Chinese version of Frost's Multidimensional Perfectionism Scale(FMPS).Methods: One thousand two hundred and eighty Chinese undergraduate students participated in the study and responded to the FMPS and instruments measuring psychological distresses such as anxiety,depression,and obsession-compulsion.Both exploratory and confirmatory factor analyses were used to examine the factor structure of the Chinese FMPS.The association between FMPS and psychological distresses was analyzed as an investigation for criterion validity.Results: Factor analyses extracted five of the original six factors of FMPS.PC did not emerge as a dimension.The five subscales were shown to have satisfactory internal consistencies with Cronbach's alpha between 0.64 and 0.81 and test-retest reliability 0.63~0.82.Chinese FMPS was associated with anxiety,depression,and obsession-compulsion.Particularly,CM and DA are correlated with these psychological distresses with coefficients between 0.30 and 0.70.Conclusion: Chinese FMPS is satisfactorily valid and reliable.It is suitable to use in Chinese college students.