Objective Up to now, the role of Brucea in early embryonic development of mice and its mechanism is still unclear.This paper aims to explore the role of Brusatol in mouse early embryonic development and its possible mechanism.Methods 100 kunming rats of clean grade(80 female rats and 20 male rats) were divided into 6 group: negative control group(no DMSO)、blank control group(culture in fresh CM with equal DMSO)、20nmol/L brusatol treated group、50nmol/L brusatol treated group、100nmol/L brusatol treated group、200nmol/L brusatol treated group(A solution of Brusatol was diluted in CM to concentrations of 20, 50, 100 or 200nmol/L.).Each group used an average of 20 embryos each time, repeated 4 times.Fertilized eggs after cultured 24h, 48h,72h, 96h were respectively 2-cell stage, 4-cell stage,morula and blastocyst stage..The embryo development rate was observed in the culture medium and the optimal concentration was selected, the embryos were collected to analysis the subcellular localization of the Nrf2 by immunofluorescence.The mRNA expression level of Cyclin B, CDK1 and the protein expression of Nrf2 were detected by Q-PCR and western blot respectively.Results In 4-cell stage, the embryo development rates of 20、50、100nmol/L brusatol treated groups[(75.0±2.8)%、(30.4±7.5)%、(4.2±5.9)%] significantly reduced compared with the negative control group[(93.0±2.8)%]、blank control group[(90.9±1.2)%].In morula stage, compared with blastocyst rates of negative control group、blank control group [(83.5±2.1)%、(84.2±1.2)%], 50nmol/L brusatol treated group[(19.3±13.1)%] decreased obviously [(79.00±0.06)% vs 100%, P<0.05].In the cellular immunofluorescence assay, the expression of Nrf2 protein in 50nmol/L brusatol treated group was lower than blank control group(P<0.05).We further found that 50nmol/L brusatol treated group decreased more mRNA levels of Cyclin B[(59.5±9.2)%] and CDK1[(56.0±1.4)%] than blank control group(100%) in G2/M phase(P<0.05).Conclusion In this study, Brusatol mainly affects the cell cycle transformation from G2 to M phase dependent on Cyclin B-CDK1, further inhibiting the development of the embryo through down-regulating Nrf2.

Expressed emotion was the describing term about the guardian of patients' emotiveness tendency to patients,which reflected attitude,tolerance degree and the coping way in crisis.It was a robust predictor of relapse of schizophrenia.Studies showed that the family intervention for the high expressed emotion guardian of patients could effectively reduce the disease relapse.By summarizing the expressed emotion's concept,assessment tools,influence factors,family interventions,the paper aimed to innovate perfect interventions and promote the patients recovery.

Objective:To explore the role of Semaphorins 3A(Sema 3A) and its receptor Neuropilin-1 (Nrp1) in the development of chronic periodontitis of rats and clinical samples.Methods:20 SD rats were divided into 2 groups.Rats in the experimental group were induced into chronic periodontitis models.Rats in control group were not treated.After 8 weeks,maxilla of all the rats were collected for micro-CT scanning and IHC staining.The distance from cementoenamel junction to alveolar bone crest(CEJ-ABC) and the IOD of Sema3A/Nrp1 positive staining in rats were analyzed.20 clinical samples of chronic periodontitis(n =10) and normal periodontal tissues (n =10) were collected for immunofluorescence and qRT-PCR analysis.Results:The CEJ-ABC distance of chronic periodontitis group was higher than that of the control group(P ＜ 0.05).The IOD of Sema3A/Nrp1 in experimental group was lower than that of the control group (P ＜ 0.05) and with a negative correlation with bone loss (P ＜ 0.05).Immunofluorescence and qRT-PCR analysis showed that the expression level of Sema3a/Nrpl in clinical samples with chronic periodontitis was also lower than that of the healthy subjects(P ＜ 0.05).Conclusion:The reduced Sema3A/Nrp1 plays an important role in the development of bone destruction in chronic periodontitis.

Objective To explore the characteristics of semantic priming effects in Chinese words with different association strength in patients with aphasia by auditory stimulation.Method Stimulus-response word pairs with different association strength including strong,moderate,weak,and no association categories were chosen from word association thesaurus as experiment materials.Both patients with aphasia(n=11)and normal subjects (n=16)were requested to finish an auditory lexical decision task for target words.Semantic priming effects were investigated by means of measuring reaction time(RT)and error rate of each word-pair.Results In patients with aphasia and normal subjects,the mean RTs were significantly shorter in strong,moderate and weak association strength words than in no association strength words(patients with aphasia(1270.20±47.70)ms,(1340.50±266.25)ms,(1429.70±317.07)ms vs(1549.00±325.87)ms,P＜0.05 and normal subjects(1140.2±274.48)ms,(1196.50±284.06)ms,(1262.10±274.31)ms vs(1391.20±315.68)ms,P＜0.05).In strong,moderate,and no association strength words,the mean RTs were no significant differences between two groups.In the weak association strength words,mean RTs were significantly longer in patients with aphasia than in normal subjects((1429.70±317.07)ms vs(1262.10±274.31)ms,P＜0.05).In two groups,mean error rates were significantly less in strong,moderate and weak association strength words than in no association strength words(patients with aphasia:7.73±6.07,4.55±7.23,6.82±8.15 vs 14.09±12.41,P＜0.05 and normal subjects:3.44±4.37,2.81±3.64,5.31±5.91 vs 10.94±11.14,P＜0.05).However,in strong association strength words,mean error rates were significantly higher in patients with aphasia than in normal subjects(7.73±6.07 vs 3.44±4.37,P＜0.05).In moderate,weak and no association strength words,there were no significant differences between two groups.Conclusion The patients with aphasia follow gradient of the association strength words like normal subjects and have semantic priming effects in the strong,moderate association strength words.

Cestode Infections ; parasitology ; Humans ; Infant ; Male

The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.

Objective To compare the mutation sites in human immunodefieiency virus type 1 (HIV-1) vpr gene via of HIV-1 infected individuals from different regions in China with the previous studies, and to provide information for the further study on the relationship between HIV-1 vpr gene mutations and clinical conditions of the patients. Methods Reverse transcription-polymerasc chain reaction (RT-PCR) and nested PCR were used to amplify HIV-1 vpr gene of 398 HIV-1 infected individuals. The amino acid sequences were analyzed to determine polymorphisms, deviation rate and common mutation sites of HIV-1 vpr gene. Meanwhile, the viral load, subsets of lymphocytes and clinical course of patients infected with mutated HIV-1 were analyzed. Results One hundred and fifty three positive samples which were obtained from 398 HIV-1 infected individuals were available for further analysis. The amino acids sequence typing of HIV-1 Vpr were showed that CRF01 AE was 51.63%, subtype C 24.84%, subtype B 17.65%, CRF03_ AB 3.92% and CRF08 BC 1.31%. Eighty four point three percent of 77th amino acid of HIV Vpr sequence was glutamic acid which was significantly different from what overseas researches reported that the R77Q mutation was correlated with long-term non-progression (LTNP) of AIDS. The mutations of the, 63th, 70th, 85th, 86th, 89th and 94th amino acids of HIV Vpr were likely related to the clinical remission of HIV-1 infected individuals. Conclusions M group is the main type of HIV Vpr typing in China, and CRF01 AE is predominant. Some amino acid mutation sites of HIV-1 Vpr are possibly correlated with clinical manifestations of HIV-1 infected individuals.

AIM: To investigate the feasibility and effect of directly differentiation of embryonic stem cells(ESC) into neural cells induced by retinoid acid(RA) without embryonic body(EB) culture period in vitro.METHODS: ESC were digested and divided into 4 groups: group A and B were undergone EB culturing.After that,cells in group A were induced by RA,cells in group B were differentiated spontaneously,cells in group C were committedly induced by RA directly without EB culturing,and cells in group D were differentiated spontaneously without EB period.Morphologic changes were observed under inverted microscope and scanning electron microscope.MAP-2 and GFAP were detected by immunocytochemistry and flow cytometry after differentiated for 9 days.RESULTS: In groups A or C,neuron-like cells increased gradually,forming neural network.At the 9th day,a large part of cells in these groups were MAP-2 positive cells,and the positive rate was higher than that in groups B or D(P0.05).CONCLUSION: ESC was directly induced into neural cells by RA without EB culture period in vitro.This modified method has the same effect as the traditional RA 4-/4+ assay and can replace the traditional method.

ObjectiveTo summarize the clinical significance of ultrasonic screening of fetal structural anomalies at 11-13+6 weeks.Methods We conducted a retrospective study of 4853 cases of nuchal translucency screening at 11-13+6 weeks in Maternal and Child Health Hospital of Bao?an of Shenzhen City from September 2011 to May 2014. The screening ultrasound planes included the median sagittal plane, neck sagittal section, cerebral transverse section, cardiac four-chamber view, three-vessel-trachea view, abdominal transverse section, bladder section, upper limb section and lower limb section of the fetuses. All the cases then underwent the ultrasonic structural screening in the second trimester (20-24 weeks) and the third (28-32 weeks) trimester and were followed up until 6 weeks after birth or the biopsy after abortion.Results Eighty-ifve fetal structural anomalies were detected among the 4853 pregnant women at 11-13+6 weeks of gestation with the detection rate of 1.75% (85/4853), including central nervous system abnormalities (28 cases), anterior abdominal wall anomalies (9 cases), cardiac anomalies (6 cases), urinary system malformation (3 cases), skeletal system malformation (2 cases), multilocular cystic tumor and dropsy embryo (35 cases), and abnormal twins (2 cases). Among above abnormal fetuses, 6 cases showed normal structure in the screening after 14 weeks and were born without malformations, while the rest 79 cases were taken artiifcial abortion (73 cases in the ifrst trimester and 6 cases in the second trimester). Only 9 cases were taken chorionic puncture or amniocentesis, including normal karyotypes (3 cases), 47, XN, +18 (3 cases) and 45, X (3 cases). The False negative rate in the ifrst trimester was 23% (25/110). Supplementary detection of fetal structural abnormalities in the second and third trimester were found in 22 cases (20%, 22/110). Two cases of VSD and 1 case of microtia were identiifed after birth.ConclusionsThe fetal malformation can be detected in the earlier gestation with the ultrasonic screening at 11-13+6 weeks, which provide the earlier termination to the abnormal fetus. It has important clinical signiifcance in effectively reducing fetal births with structural abnormalities.

0.05).Conclusion CYPIA1 MspⅠ polymorphism is not related to the susceptibility to colorectal carcinoma.