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.

Astragalus adsurgens seed is one of the most common adulterants of Astragali Complanati Semen in the market, whose morphological characteristics are very similar with A. complanatus seeds (Astragali Complanati Semen). Many identification methods have been reported, such as morphological identification, fluorescence method, ultraviolet spectrophotometry, TLC, HPLC, protein electrophoresis and so on, but there's no much about microscopic identification. In the present study, the morphological characteristics and microscopic characteristics of these two seeds were investigated, which could provide scientific evidence for the identification and classification of Astragali Complanati Semen. Our results showed that these two seeds were slightly different in the color and the appearance, but significantly different in the microstructure of the seed coat and the hilum, and the distribution of fat droplets in the cotyledon cells. So these microscopic characteristics can be applied for the identification of Astragali Complanati Semen.
Astragalus Plant ; anatomy & histology ; chemistry ; Color ; Discriminant Analysis ; Microscopy ; methods ; Quality Control ; Seeds ; anatomy & histology ; chemistry

Akebia trifoliate has been reported to have many pharmacological activities and the roots, petioles and seeds are used to different symptoms. However, the structure and anatomy of its seeds was almost not reported until now. In the present study, we investigated the morphological characters of the fruit and seed, and the anatomical characters of the testa, micropyle, embryo and endosperm, which could provide evidences for the study on classification, identification and application of A. trifoliate. Our results showed that the testa of A. trifoliate consisted of an epidermic cell layer, the sclerenchyma cells layer, the parenchyma cells layer and an innermost pigment layer. At the micropylar region, the outermost epidermal cells were specialized the white caruncle-like structure and the testa included a lot of lignified tissues. Endosperm comprises two layer cells. Outermost yellowish-brown layer cells contains lots of fat droplets, and innermost white layer cells contains lots of aleurone grains and crystalloids.
Germination ; Magnoliopsida ; anatomy & histology ; growth & development ; Seeds ; anatomy & histology ; growth & development

Aged ; CDX2 Transcription Factor ; Cysts ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Homeodomain Proteins ; metabolism ; Humans ; Keratin-20 ; metabolism ; Mucous Membrane ; pathology ; Peritoneal Neoplasms ; metabolism ; pathology ; surgery ; Pseudomyxoma Peritonei ; metabolism ; pathology ; surgery ; Rupture ; Splenic Neoplasms ; metabolism ; pathology ; surgery

OBJECTIVETo discuss arthroscopic technique of single Kirschner wire and suture avoiding epiphyseal line fixation for tibial intercondylar eminence fracture and its clinical results.

METHODSFrom May 2008 to December 2012, 21 patients (13 males, 8 females, ranging in age from 6 to 14 years old) with tibial intercondylar eminence fracture were treated arthroscopically with single Kirschner wire and suture avoiding epiphyseal line fixation technique. According to Meyers and McKeever classification, 7 patients were type II, 10 patients were type III, and 4 patients were type IV. Active rehabilitation began at one week after operation. The patients were followed up for 10 to 30 months. X-ray films were taken to evaluate fracture healing at 1, 3, 6 months after operation; range of motion, the anterior drawer test, the lachman test and the Lysholm knee score were used to evaluate clinical effects.

RESULTSAll fractures were healed without displacement at 6 weeks after operation. Anterior drawer test and the lachman test were both negative in all patients at 3 months after operation. Lysholm knee score was 95.5 ±2.5 at 6 months after operation, and postoperative X-ray film did not find epiphyseal line broadening or narrowing.

CONCLUSIONArthroscopic treatment for tibial eminence intercondylar fracture with single Kirschner wire and 8-shaped suture avoiding epiphyseal line fixation technique has many advantages, such as firm fixation,early mobilization, less invasive, less injury of physis and satisfactory effect.

Adolescent ; Arthroscopy ; Bone Wires ; Child ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Sutures ; Tibial Fractures ; surgery

Animals ; Electromagnetic Fields ; Embryo, Mammalian ; metabolism ; Female ; Mice ; Pregnancy ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

In this study, ITS2 barcode was used to identify Bupleurum chinense and B. longiradiatum. The ITS2 regions of 48 samples were amplified and sequenced. The sequences obtained above were aligned and the K2P distances were calculated. We used three methods, BLAST1, nearest distance and phylogenetic tree (NJ-tree), to test the identification ability. The results showed that the maximum intraspecific genetic distance of B. chinense was 0.013, and the minimum interspecific genetic distance between B. chinense and B. longiradiatum was 0.049. The NJ-tree can easily identify B. chinense and B. longiradiatum. Therefore, the ITS2 barcode is suitable to identify B. chinense and B. longiradiatum.
Bupleurum ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control

Objective To monitor the dynamic changes of liver and kidney functions in the first week after liver transplantation and to assess the value of liver and kidney functions in predicting patient's survival. Methods clinical data of 161 recipients with benign liver diseases were retrospectively reviewed.Total bilirubin ( TB ), aminoleucine transferase ( ALT), aspartate aminotransferase ( AST), prothrombin time (PT) and serum creatinine (SCr) were recorded and analyzed during the first post-transplant week.The data collected at dl were analyzed in the multivariate COX regression. Results From d1 to d7 post-transplant, the median value changed from 116. 2 mmol/L to 66. 7 mmol/L ( Z = 5.901, P < 0. 01 ) for TB,19.4 s to 15.0 s (Z = 11. 657, P <0.01 ) for PT, 285 U/L to 100 U/L (Z = 12.619, P<0. 01 ) for ALT,264 U/L to 50 U/L (Z =9. 776, P <0. 01 ) for AST, 103.4 mmol/L to 86. 6 mmol/L (Z = 1. 353, P 0. 05 ) for SCr. Post-transplant SCr ( RR = 3. 477, P < 0. 001 ) and TB ( RR = 2. 088, P < 0. 05) levels wereindependent factors influencing patient survival. A prognostic score formula was then established as 1. 276 ×InSCr (mg/dL) + 0. 730 × InTB (mg/dL). Conclusion In a successful liver transplantation,transplanted liver function recovers promtly within one week. SCr and TB levels on post-transplant d1 have good prognostic value.

Objective　To investigate the relationship betwee n tricuspid insufficiency (TI) and the function of the right ventricle, degree o f pulmonary vascular pathological change in patients with rheumatic heart diseas e so as to provide the indication of operation for treating TI. Methods 　41 cases of rheumatic heart disease with TI accompanying with bicuspid pathological changes were reviewed. Correlation analysis was done between the s everity of TI and the right ventricular size(RVS), pulmonary artery pressure(PAP ), pulmonary artery resistance(PAR) etc. Results　It was found t hat the size of the right ventricle, PAP, PAR were positively correlated to th e degree of TI. Tricuspid annuloplasty should be carried out in patients with RV D>40 mm, PAR >48 kPa*s/L, PAP>8 kPa. Conclusion　TI resulted ma inly from insufficient function of the right ventricle and marked patholog ical changes of the pulmonary blood vessels. Doppler echocardiography evaluation for the function of right ventricle and pathological condition of pulmonary blo od vessels might be of significant in deciding whether tricuspid annuloplasty sh ould be performed simutaneously in patients of bicuspid valve replacement.

Animals ; Cell Hypoxia ; Cells, Cultured ; Chemokine CXCL12 ; genetics ; metabolism ; Endothelial Cells ; cytology ; metabolism ; Female ; Gene Expression Regulation ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Immunohistochemistry ; Male ; Pulmonary Artery ; cytology ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine