Purpose The aim of this study was to reveal the role of SGT,the small glutamine-rich tetratricopeptide repeat (TPR)-containing protein,in the developing mouse brain through examining the expression profile of SGT during the development stage. Methods In this study, quantitative RT-RCR and Western blot were applied to investigate the expression of SGT mRNA and protein in the mouse whole brain. Western bolt was also used to detect the expression of SGT in the hippocampus, cerebral cortex, striatum and cerebellum. Immunohistochemical analysis on postnatal and adult mouse brain was performed to examine the subcellular localization of SGT. Results Our data showed that the levels of SGT mRNA and protein in the mouse whole brain were both high during the postnatal stage and declined in the adult. Regional expression of SGT protein in the hippocampus, cerebral cortex, striatum and cerebellum showed a similar expression profile.Immunohistochemical analysis found that in the P14 mouse brain, SGT was abundant in all the CA regions of hippocampus as well as most regions of cerebral cortex and striatum. In the cerebellum, SGT was mainly distributed in Purkinje cells. In the sections of the adult mouse brain, faint expression was observed in the regions mentioned above. Conclusions Our findings firstly exhibit the expression pattern of SGT in the mouse brain development,which might shed new light on further functional analysis of SGT in the central nervous system.

Objective To evaluate the clinical efficiency of intermittent photic stimulation (IPS) with pattern and red color plastic plates in video-electroencephalography(EEG).Methods Three hundred and fifty-eight patients hospitalized in Wuhan Children's Hospital from March 2013 to March 2014 were selected,and they were examined by using stroboscope with a red plastic plate,a dots printed plastic plate,and ordinary white flicker for photic driving response (PDR),photoparoxysmal responses (PPR),photoconvulsive response (PCR).The results of patients with PDR,PPR,PCR were analyzed.Results The PDR synchronously evoked by the three flickering patterns were in 29 cases,and the amplitude of photic drivings evoked by ordinary white flicker and red flicker were higher than that of flickering dot pattern [(30.294 ± 7.767) μV,(31.103 ± 8.920) μV vs (24.436 ± 8.075) μV],and there were significant differences(t =2.983,2.815,P =0.003,0.008).However,there was no significant difference between ordinary white flicker and red flicker (t =0.368,P =0.710).The PPR evoked were in 17 cases,and 16 cases (94.12％) of them were evoked by red color,while 10 cases (58.82％) wcre evoked by white light,and 1 case only evoked by flickering dot pattern.However,stronger intensity and longer duration time of PPR were evoked by red color than by white light.In 3 patients with positive PCR,the epileptic seizure was more likely to be evoked by red color or pattern plastic plate.Conclusions IPS with red and pattem plastic plate is more potent in eliciting photosensitive epilepsy in video-EEG than the ordinary white light.Moreover,it could reduce the discomfort of eyes without influencing PDR in comparison to the ordinary white light.

Objective:To study the amino acid and codon usage profile of HIV-1 Env gene in donors whose serum exhibit highly broad cross-neutralizing activity. Methods:The samples were divided into highly broad cross-neutralizing activity group (hBCN + group) and non-highly broad cross-neutralizing activity group (hBCN - group) based on whether the neutralization breadth was higher than 90% or not. Full-length Env genes were amplified by single genome amplification (SGA) method from patients′ plasma samples, and the characteristics of Env sequences in hBCN + group were compared with hBCN - group. The correspondence analysis (COA) on relative amino acid usage (RAAU), adaptability to host based on similarity index D( A, B) and relative synonymous codon usage (RSCU) values of Env genes (hBCN + and hBCN -) with respect to human host RSCU were analyzed. Results:Correspondence analysis showed that the RAAU data of hBCN + group and hBCN - group were distributed along the two main axes to form two relatively separated clusters, indicating that the Env genes of the two groups had relatively unique amino acid usage patterns; the similarity index calculation results showed that hBCN + group (0.097) was lower than the hBCN - group (0.102), in addition, the Env gene of the hBCN + group had less frequency of similarly selected codons with human host system compared to hBCN - group. Conclusions:Env genes in hBCN + group and hBCN - group may have relatively unique amino acid usage patterns, and virus strains in hBCN + group are less adaptable to the host than those in hBCN - group.

Objective To explore the teaching effects of Lecture Based Learning (LBL) incorporated with Case Based Learning (CBL) and Problem Based Learning (PBL) methods in the teach-ing of anatomy.Methods 740 students of Grades 2012 and 2013 from clinical medical major were chosen as the subjects,randomly divided into experimental and control groups.LBL was conducted in the control group of 644 students,while LBL+PBL+CBL method was applied in the experimental group of 96 students.After completion of the course,all students were surveyed with the questionnaire and tested with specimen and theoretical examinations.The data was collected and analyzed with SPSS 17.0 and two sample U tests.Results The students' time of learning anatomy in the control group outside classroom teaching was (105 ± 25) minutes per week,while the students' time of learning anatomy in the experimental group was (190 ± 25) minutes per week.The difference was statistically significant (t=31.08,P=0.000).The difference of after-class total learn-ing time was also statistically significant between two groups(t=27.42,P=0.000).Percentages of satisfaction with teaching methods for control group were 87.5％ and 87％,while for the experimental group 98％ and 100％.Kruskal-Wallis test showed there were significant differences in experimental groups of grades 2012(x2=31.19,P=0.000) and 2013 (x2=40.35,P=0.000) compared with control group.Comparison of both groups' results of the examinations was statistically significant concerning multiple choices,essays and case analysis questions (P＜0.05).Conclusion Teaching method of LBL incorporated with PBL and CBL was significantly effective than that of the sole traditional LBL method,which indicated its practice value.

Objective:To analyze the chicken egg-white extracts were co-cultured with cells whether elevated stem cells protein,whether the cells transformation into stem cell.Methods:Four kinds of cells,making a common culture,a 50% chicken egg-white extract co-cultured for 3 days,cells were collected and frozen at -80 degrees,sending the company to do stem cell protein microarray.Results:C57-BMSC has three proteins occurred statistically significant change , TS-UC-MSC has one proteins occurred a statistically significant change ,293T has one protein occurred a statistically significant change ,and 293T-GFP has one protein occurred a statistically significant change.Conclusion:50% chicken egg-white extract co-cultured cells,the cells occurred the phenomenon of transformation into stem cells.

BACKGROUND:Systemic lupus erythematosus is an autoimmune disease characterized as an emergence of a variety of autoantibodies in serum and multi-system and multi-organ lesions. Currently, there is a lack of effective treatment options, and umbilical cord mesenchymal stem cel s are a promising therapy for systemic lupus erythematosus based on cel biological roles. OBJECTIVE:To observe the therapeutic efficacy of human umbilical cord mesenchymal stem cel transplantation in the treatment of systemic lupus erythematosus in mice. METHODS:Human umbilical cord mesenchymal stem cel s were isolated and cultured fol owed by labeling with DiR fluorescence. Experimental mice were divided into normal control group (C57BL mice), model control group (C57BL/lpr mice), low-, medium-and high-dose umbilical cord mesenchymal stem cel therapy groups (C57BL/lpr mice), with 10 mice in each group. Mice in the low-, medium-and high-dose groups were respectively injected 0.5×106, 1×106, 2×106 human umbilical cord mesenchymal stem cel s, once a week, for 3 consecutive weeks. At the end of treatment, blood samples were col ected to measure antinuclear antibody, anti-histone antibody, anti-double stranded DNA antibody changes;OPG and Foxp3 gene expression changes were detected by quantitative PCR method. RESULTS AND CONCLUSION:After treatment, the levels of anti-nuclear antibodies, anti-histone antibodies and anti-double stranded DNA antibodies in the peripheral blood of mice were al declined in the low-, medium-and high-dose groups, while the number of peripheral blood CD4+CD25+T cel s was significantly elevated. OPG and Foxp3 gene expression was also increased dramatical y in the low-, medium-and high-dose groups, which was similar to that in the normal control group and significantly different from that in the model control group (P<0.01). Experimental findings demonstrate that after transplantation of human umbilical cord mesenchymal stem cel s, al relevant indicators in C57BL/lpr mice recovered to the normal levels, and the high-dose treatment group had the most obvious effect.

Objective To compare the parameters of bilateral tubal ligation by LESS and multi-incision laparoscopy and their prognosis of IVF-ET. Methods From Jun. 2016 to Dec. 2017,32 patients underwent LESS bilateral tubal ligation and 60 patients under-went multi-incision laparoscopic bilateral tubal ligation were enrolled. The demographic data ,operative parameters ,and postoperative prognosis of IVF- ET in the two groups were summarized and compared. Results The mean operating time in LESS group was prolonged and its operative hospital stay was significantly decreased compared to multi-incision laparoscopic group. No significant difference was observed in the intraoperative blood loss in two groups. Moreover,no significant differences were found in the AFC,AMH,Gn stimulation days,total Gn dosage,Ovum number,good quality embryo rate,embryo implantion rate,clinical pregnancy rate and early spontaneous abortion rate in two groups. Conclusions For experienced laparoscopic surgeons,LESS is a safe and feasible procedure with similar perioperative outcomes to multi-incision laparoscopic surgery.

Objective To observe the effect of different explosion impulse on rabbit lung injury and decide the death curve,so as to provide a reference for the prediction of lung injury.Methods Six healthy male New Zealand white rabbits with weight of 2.0-2.5 kg and age of (6 ± 1)months were selected.The rabbits were put 0.5 m,0.6 m,0.7 m,0.9 m,1.0 m,and 1.2 m away from 90 g TNT to carry out the blast injury experiment.The characteristic parameters of blast shock wave and general lung injury were recorded.Based on the experimental results combined with theoretical analysis,the changes of rabbit lung injury depending on the explosion distance as well as the rabbit death curve were determined.Results After the 90 g TNT explosion,the peak overpressure of shock wave and the corresponding specific impulse decreased quickly with the increase of explosion distance.The peak overpressure was 0.79 MPa and the specific impulse was 82 Pa · s at the explosion distance of 0.5 m.The peak overpressure was 0.1 MPa and the specific impulse was 34 Pa · s at the explosion distance of 1.2 m.The rabbits at 0.5 m and 0.6 m died,the rabbit at 0.7 m was severely injured,and the rabbits at 0.9 m,1.0 m,and 1.2 m were slightly injured.The dependence of lung injury degree on the explosion distance under 90 g TNT explosion was established based on dimensional analysis theory.The lung injury degree was exponentially attenuated with the explosion distance:φ =(R/0.6)-5.64(φ represented lung injury degree,and R represented the explosion distance).Considering the combined injury effects of peak overpressure of shock wave and its specific impulse on rabbit lung,the death curve of rabbit was determined:(p-0.1) (I-59) =2.6 (p represented peak overpressure,and I represented specific impulse).The criterion of "overpressure-specific impulse" was used to estimate the death of rabbit,and the death curve of rabbit was determined as (p-0.1)(I-59) =2.6(p represented peak overpressure and I represented specific impulse).The critical overpressure was 0.1 MPa and the critical specific impulse was 59 Pa · s.Conclusions Under the explosion condition of 90 g TNT,the relationship between degree of lung injury in rabbits and explosion distance is established.Death curve of rabbits is determined based on the damage effect of shock wave peak overpressure and specific impulse on the lungs of rabbits,which is significant for predicting the blast injury.

The experiment was designed to investigate the function of SREBP cleavage-activating protein (SCAP) mutant (D443N) by constructing an eukaryotic expressive vector using a smooth muscle specific promoter SM22 (pGL3-SM22-SCAP(D443N)). SM22 promoter (pSM22) was amplified from genome DNA of mice by nested PCR, and then cloned into pMD-T vector. The SM22 promoter fragment released from the vector by Kpn I and Hind III digestion was sub-cloned into pGL3-control-Luc vector, to form pGL3-SM22-Luc. The activity of pSM22 in human vascular smooth muscle cells (VSMCs) was tested using Dual-Luciferase Reporter System. SCAP(D443) mutant amplified from plasmid pTK-HSV-SCAP(D443N) and pSM22 from mice liver were cloned into pGL3-control vector to construct pGL3-SM22-SCAP(D443N) which was transfected into Chinese hamster ovary cells (CHO) to test SCAP(D443) expression by real-time PCR and Western blot. The sequence and construction of pGL3-SM22-SCAP(D443N) were correct. SM22 promoter activity initiated the expression of luciferase in VSMCs and also drove SCAP(D443) expression in transfected CHO cells. The pGL3-SM22-SCAP(D443N) eukaryotic expression vector was successfully constructed and the recombinant vector provides a powerful approach in investigating the function and regulation of SCAP and also in producing vascular smooth muscle specific SCAP transgenic mice.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Genetic Vectors ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; physiology ; Membrane Proteins ; biosynthesis ; genetics ; physiology ; Mice ; Mice, Transgenic ; Microfilament Proteins ; genetics ; Muscle Proteins ; genetics ; Mutant Proteins ; biosynthesis ; genetics ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

Objective To retrospectively analyze the clinical and laboratory characteristics of patients with positive blood culture results for Mycobacterium tuberculosis (M.tb).Methods The clinical laboratory database of patients suspected with disseminated tuberculosis from January 2009 to January 2017 in Huashan Hospital affiliated with Fudan University were collected and analyzed.The clinical manifestations,laboratory characteristics and outcomes between disseminated tuberculosis patients with positive blood culture (positive blood culture group) for M.tb and negative results (negative blood culture group) were compared.T test,Mann-Whitney U test and Fisher exact test were used for statistical analysis.Results A total of 5 589 patients suspected with M.tb infection had peripheral blood culture for mycobacterium.Positive blood culture for M.tb was found in 26 disseminated tuberculosis patients,while 6 patients finally identified as nontuberculous mycobacterium (NTM) with species identification,and 22 disseminated tuberculosis patients with negative blood culture results were enrolled during the same period as control.The mean age ([49.1 ± 10.1] years old vs [38.3 ± 17.1] years old,t =2.460,P =0.018),the proportion of diagnosed with fever of unknown origin at admission (FUO) (65.0％ [13/20] vs 13.6％ [3/22],P =0.001),the proportion of diagnosed with focal infection (30.0％ [6/20] vs 86.4％ [19/22],P =0.001),the proportion of patients with other diseases (75.0％[15/20] vs 22.7％ [5/22],P =0.002),the proportion of patients with hematological diseases (35.0％ [7/20] vs 4.5％ [1/22],P =0.018) and the proportion of patients with tumor (20％ [4/20] vs 0[0/22],P =0.043) in the positive blood culture group were significantly different from those in the negative blood culture group.Laboratory examinations of the percentage of neutrophils,the percentage of lymphocytes,the percentage of monocytes,the value of neutrophil/lymphocyte,the level of hemoglobin,the level of erythrocyte sedimentation rate,the level of C-reactive protein,the level of procalcitonin and the positive rate of T-SPOT.TB in positive blood culture groups were significantly different from those in negative blood culture group (all P ＜ 0.05).Conclusions Peripheral blood M.tb culture is more likely to be positive for those elder disseminated tuberculosis patients with hematological diseases or tumors,and those with increase of neutrophil counts and inflammation markers but reduction of lymphocyte counts and hemoglobin.