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.

BACKGROUND: Embryonic stem cells (ESCs), the seed cells of all mature cells in vivo, are useful tools for nerve transplantation and developmental gene function research. Notch1 signaling pathway is the key pathway to control the ordered neural development and differentiation of many kinds of neural cells, however, there is no report on the dynamic expression of Notch1 signal during the ESC differentiation to date. OBJECTIVE: To investigate the expression of Notch1 protein, transmembrane signal transduction molecule, during directional differentiation of embryonic stem cells into neural cells. DESIGN, TIME AND SETTING: Cell research was carried out between October 2003 and October 2004 at Zhongshan Ophthalmic Center, SUN Yat-sen University, Guangzhou, Guangdong Province, China. MATERIALS: BALB/C mouse embryonic stem cell line Ⅵ (passage 11)was obtained from experimental animal center of SUN Yat-sen University, provided by professor Huang Bing. ESC culture medium was high-glucose DMEM medium with 20% bovine serum and 106 IU/L mouse leukemia inhibitory factor. Induced differentiation medium was high-glucose DMEM medium with 20% fetal bovine.serum and 5×107 mol/L retinoid acid(RA). METHODS: Passage 11 ESCs were resuscitated and incubated by ESC culture medium in incubator at 37℃ with 5% CO2. Passage 11 ESCs were subcultured after 2 or 3 days and RA was added into medium to induce differentiation. Three time points for observation were established: induced for 1, 5 and 9 days. MAIN OUTCOME MEASURES: Morphological changes were observed under inverted phase contrast microscope, MAP-2 antigen expressed in differentiated cells was detected by immunofluorescence method. Immunocytochemistry, Western Blot, flow cytometry assay were used to investigate the Notch1 protein expression. RESULTS: ESCs presented clone-like growth. After induced by RA for 9 days, single neural network was achieved around most of the cell clusters. With the prolongation of induction, MAP-2 positive neural cells increased gradually. Almost all ESC clones expressed Notch1 protein strongly or positively, but Notehl protein expression decreased gradually after induced differentiation (P < 0.01). CONCLUSION: Notch1 signal shuts off progressively during induction of ESCs into neural cells, which suggests Notch1 may play an important role in the differentiation of ESCs into neural cells.

A strain of thermotolerant halophilic bacteria YJ0238 was isolated from the salt well at 47℃ in the Kangning countryside where was located at the side of Lancang River in the Tibet Autonomous Region. Some physiological and biochemical properties were characterized.16S rRNA gene sequence of YJ0238 was amplified by PCR,and its nucleotide sequence was determined. Based on it’s physiological and biochemical properties,homology and phylogenetic analysis of 16S rRNA gene sequence,strain YJ0238 was identified as a subspecies of the species Idiomarina zobellii. The GenBank accession number of the 16S rRNA gene sequence of strain YJ0238 is EF693953. Until now,there were few reports on the study of high-temperature and high-salt microbial in domestics. The results of this study will provide research material and information for further studies in this area.

OBJECTIVETo explore the essence of Spleen-Kidney Deficiency in middle-aged patients.

METHODSInvestigation was carried out in 773 cases of 50-69 years old to assay 34 parameters in them, including blood lipid, oxidation and anti-oxidation related substance, sex hormone, liver and renal function, immune function, blood routine, blood pressure and lung vital capacity, etc.

RESULTSSpleen Deficiency Syndrome is closely related with lipid metabolism disorder; Kidney-Qi Deficiency Syndrome is closely related with hypoimmune function; Kidney-Yin Deficiency Syndrome is closely related with lipid metabolism disorder and hypertension, and Kidney Yang Deficiency is closely related with weakness of anti-oxidation capacity, hypoimmunity, the internal environment disorder of sex hormone and aging, manifested as multiple functions abating.

CONCLUSIONSpleen Deficiency, Kidney-Qi Deficiency and Kidney Yang Deficiency are different layers of a gradually developed and aggravated pathological process, but Kidney-Yin Deficiency could not be listed into this layer.

Aged ; Aging ; CD4-CD8 Ratio ; Cholesterol ; blood ; Diagnosis, Differential ; Female ; Humans ; Kidney Diseases ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Splenic Diseases ; blood ; Superoxide Dismutase ; blood ; Yang Deficiency ; blood ; Yin Deficiency ; blood

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

Objective To evaluate the clinical significance of fetal nasal bone absence and thickened nuchal translucency ( NT) at 11-13 +6 weeks ultrasound screening .Methods A total of 4200 pregnant women with single fetus registered at Mother and Children ’ s Health Care Center in our hospital were examined at 11-13 +6 gestational weeks .Both fetal nasal bone and NT ultrasound evaluation were offered to assess whether nasal bone is absent and NT is thickened (>3.0 mm) in these cases.Particular attention was paid to the relationship between abnormal findings ,karyotype and pregnancy outcome .Results In all, 3492/4200 cases were included in the study with both NT measurement and nasal bone evaluation .Seven hundred and night cases were excluded because of unavailable clinical outcome .Among 3492 fetuses:(1) There were 3 cases absent of nasal bone .Among the 3 cases without nasal bone , 2 cases ( 1 case combined with thickened NT ) were trisomy 21(66.7%,2/3).(2) There were 351 cases with NT>3.0 mm (10.1%,351/3492).Among the 351 cases with thickened NT,there were 4 with trisomy 21 syndromes (1.14%,4/351,1 case combined with nasal bone absence ),1 with trisomy 18 syndrome,1 with Turner syndrome,6 with structural anomalies but normal karyotype (1.71%,6/351).(3)Among the 3139 cases with normal nasal bone and NT ,there were 8 cases with chromosomal or structural anomalies .Conclusions Absent nasal bone and thickened NT are important markers of trisomy 21 in the first trimester ultrasound screening .Thickened NT has significant correlation with other fetal chromosomal and structural anomalies .

Objective To evaluate the value of stereotactic core needle biopsy(SCNB)in diagnosis of breast lesions.Methods Forty-seven cases were punctured with computer-assisted stereotactic system, spring-loaded biopsy guns and 16 G core needles.The record of each item was collected,including clinical manifestations,descriptions of the mammographic characteristics(such as calcification,mass and architectural distortion),the pathology of the SCNB and the surgical pathology or mammographic follow-up data.Then the results of SCNB were analyzed based on the comparison of SCNB pathology and the surgical pathology.The reason that SCNB failed and misdioagnosed was inferred from the relationship of SCNB accuracy and the X-ray characteristics.Results Forty-four cases were punctured successfully,3 cases failed.Thirty-one patients were operated soon after biopsy.The results of 27 SCNB cases agreed well with the final pathology but the other 4 did not.Conclusions SCNB as an accurate,time-saving and cost- effective method,is also minimally invasive and hardly changes the architecture of the breast.SCNB can diagnose breast lesions in advance,reduces the number of surgical biopsy,and is promising in clinical application.

Objective To improve the anderstanding of clinical profile of primary plasma cell leukemia.Methods Case report and literature review.Results A rare case of nervous system relapse in primary plasma cell leukemia was reported.Six patients were identified from the literature.The type of immunoglobulin included IgG(3 patients),IgD(2 patients).Clinical manifestations of nervous system were variable.The average interval from initial diagnosis to the development of nervous system relapse was 16.5 months.Plasma cells were found in cerebrospinal fluid in 4 patients.The mean surviaval time was 6.7 months after nervous system relapse.Conlusion Nervous system relapse in primary plasma cell leukemia is rare with poor prognosis.

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.

This article discussed on the continuing medical education for general practitioners within a nonprofit multihospital system.Literature review,statistical analysis and survey results were employed.Training situation for general practitioner at home and aboard was reviewed.Statistics on population,disease spectrum,present situation of community doctors in Luwan branch of Ruijin hospital was also analyzed.Results showed that current education system for general practitioners cannot catch up the increasing demand yet.Meanwhile patients expected well qualified and skillful physicians in the community healthcare centers.Reinforcement of continuing medical education for general practitioners was the stepping stone of a nonprofit multihospital system.Improving skills and qualifications of physician played an essential role in the urban network of medical services.