Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.
Biomass ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Culture Media ; chemistry ; metabolism ; Eriobotrya ; chemistry ; growth & development ; metabolism ; Triterpenes ; analysis ; metabolism

Objective To explore the relationship between X - linked spondyloepiphyseal dysplasia tarda (SEDL) gene escaping X chromosome inactivation( XCI) and SEDL phenotype. Methods RT - PCR was performed on total RNA which was isolated from blood samples of patients, female carriers and controls. Patients and female carriers were selected from the pedigree with SEDL caused by the mutation (IVS2 - 2A→C) of the gene. cDNA was analyzed by polyacrylamide gelelectrophoresis(PAGE). Results PAGE data indicateed that female carriers expressed both normal and mutant SEDL mRNA,meaning the SEDL gene escaping XCI. Family investigation showed carrier females in the SEDL pedigree presented no symptoms. Conclusions The SEDL gene escaping X chromosome in-activation is firstly identified from human body. This may explain that carrier females present no symptoms.

Breast Neoplasms ; diagnosis ; pathology ; Female ; Humans ; Leiomyosarcoma ; diagnosis ; pathology ; Middle Aged ; Skin Neoplasms ; pathology ; secondary

Background Human retinal pigment epithelial (RPE) cell transplantation treating retinal degenerative diseases is a researching topic,and the source of human RPE cells is a key problem.Many biological carriers can be used for the preparation of RPE cell layer.However,some advantages,such as cytotoxicity,lack of stability and immunologic reaction etc.are still existed.To study an ideal biological carrier is very important.Objective This experimental was to determine the effects of amniotic membrane on the proliferation and differentiation of human RPE cells and the possibility as a scaffold for RPE cell transplantation.Methods ARPE19 cell line cells were cultured and passaged in DMEM/F12 medium with 10％ fetal bovine serum,and 8-12generation of cells were used.The cells were divided into two groups.One group of cells were incubated on the denuded amniotic membrane,and the other group of cells were cultured in the medium (control group).MTT was performed to detect the A492 value of RPE cells for the evaluation of cell proliferation ability 24,48,72,96 hours after culture.Cell morphology was compared by histopathological examination 3 weeks after culture.The mRNA expression of pigment epithelium-derived factor (PEDF),N-cadherin,β-catenin and cell connection related proteins in the cells of both groups were assayed using reverse transcription polymerase chain reaction (RT-PCR).Ultrastructure of the cells was observed under the transmission and scan electronic microscope 3 weeks after culture.Results The number of ARPE-19 cells cultured on denuded amniotic membrane was decreased significantly in comparison with the normal culture plate(F=41.760,P =0.000).Histopatholy also showed that the cell density on amniotic membrane was lower than of normal cells on plate surface.Moreover,the expression level of claudin 1 mRNA,N-cadherin mRNA and PEDF mRNA were significantly up-regulated in denuded amniotic membrane group in comparison with control group (t=15.828,P=0.000 ;t=6.839,P=0.002 ;t=14.667,P=0.000),but the expression of Connexin 43 mRNA was down-regulated in denuded amniotic membrane group compared with control group(t=3.358,P=0.024).Ultrastructural examination revealed that ARPE-19 cells cultured on amniotic membrane exhibited a polygonal epithelial phenotype with cilium on the apical side,however,the cells cultured on normal culture plate displayed fusiform shape and uneven thickness.Conclusions Amniotic membrane plays a promoting effect on the differentiation of ARPE-19 cells and a inhibitory effect on the proliferation of ARPE-19 cells,suggesting that amniotic membrane might be an useful scaffold for the preparation of functionally mature RPE cells for clinical transplantation.

Antigens, CD20 ; metabolism ; Antigens, CD34 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; Male ; Melanoma ; metabolism ; pathology ; Middle Aged ; Prednisone ; therapeutic use ; Vascular Neoplasms ; drug therapy ; metabolism ; pathology ; Vincristine ; therapeutic use

BACKGROUND:Intertrochanteric fracture can obtain good curative effects after active effective internal fixation in early stage.
OBJECTIVE:To compare dynamic hip screw+trochanter stabilizing plate and Gamma nail fixation for treatment of Evans type IIIB and type IV intertrochanteric fracture.
METHODS:A total of 67 patients with Evans type IIIB and type IV intertrochanteric fracture, including 28 cases undergoing dynamic hip screw+trochanter stabilizing plate and 39 cases undergoing Gamma nail, were selected from Chongming Branch, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from February 2009 to November 2012. Al patients received antibiotics and functional exercises after treatment. Time of therapy, intraoperative bleeding amount, fracture healing time and Harris Hip Score were retrospectively analyzed in patients of both groups.
RESULTS AND CONCLUSION:Al patients were fol owed up for 6-45 months, averagely 21 months. (1) One patient affected infection in the dynamic hip screw+trochanter stabilizing plate group. He was healed after fixation removal half a year after drug exchange. One obese patient experienced injury again during out-of-bed activity at 3 weeks after treatment in the Gamma nail group. Fracture appeared at distal end of lock pin, and healed after replaced by Gamma nail. Trochanteric stabilizing plate was unstable in four patients of the Gamma nail group. They affected severe pneumonia after over 6 weeks of lying in bed, and healed after treatment in the medical department (2) Coxa vara appeared in two cases of the dynamic hip screw+trochanter stabilizing plate group, and three cases in the Gamma nail group. Fracture healing was achieved in al patients. (3) The time of therapy was longer, and intraoperative bleeding amount was significantly more in the dynamic hip screw+trochanter stabilizing plate group compared with the Gamma nail group (P<0.05). No significant difference in healing time and Harris Hip Score was detected between the two groups (P>0.05). Dynamic hip screw+trochanter stabilizing plate and Gamma nail are effective methods for treatment of Evans type IIIB and type IV intertrochanteric fracture. To choose a suitable fixator, we should aggregately analyze fracture stability, healing state and the degree of osteoporosis.

Anion Exchange Protein 1, Erythrocyte ; metabolism ; Breast ; metabolism ; pathology ; surgery ; Cadherins ; metabolism ; Female ; Fibrocystic Breast Disease ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Middle Aged ; S100 Proteins ; metabolism

Adult ; Eccrine Glands ; pathology ; Female ; Hamartoma ; pathology ; Humans ; Sweat Gland Diseases ; pathology

OBJECTIVETo investigate the early diagnosis and treatment for trauma around the knee with popliteal vascular injury.

METHODSA retrospective analysis was employed to analyze the clinical data from 15 patients (9 males and 6 females were with a mean age of 39.2 years old,ranging from 26 to 62 years old) with fracture or dislocation around the knee with popliteal vascular injury from January 2007 to January 2013. Combined with clinical symptoms and signs, oxygen saturation monitors, color ultrasound, DSA angiography and interventional surgery were used to determine the vascular injury. The knee fracture and dislocation were fixed with hybrid external fixation and plate-screw fixation, respectively. Then, the blood circulation was reconstructed by thrombectomy, repair and autologous vein graft for individual injured vascular. The average total operation time, average hospitalization days, predictive salvage index (PSI), average blood transfusion amount, average medical expenses and infection cases were recorded to determine the effect of early diagnosis and treatment.

RESULTSThere was one patient with death, 8 patients with amputation, and 6 patients with successful repair surgery for popliteal artery, anterior tibial and posterior tibial arteries. These six patients with surviving limbs were followed up for an average of 28.3 months (ranged, 12 to 60 months). Among the 6 successful patients, the joint function of 4 patients was good and excellent.

CONCLUSIONThe trauma around the knee with popliteal vascular injury is characterized by complex and serious injury, easy misdiagnosis and loss diagnosis, poor prognosis and high risk of amputation. The early diagnosis of trauma around the knee with popliteal vascular injury should depend on the mechanism of trauma, local anatomical characteristics of injury site, clinical presentations and appropriate auxiliary examinations. The appropriate indications for limb salvage and amputation should be used to achieve more effective clinical results.

Adult ; Early Diagnosis ; Female ; Humans ; Knee Injuries ; diagnosis ; surgery ; Male ; Middle Aged ; Popliteal Artery ; injuries ; surgery ; Retrospective Studies

Cloning, Molecular ; DNA ; genetics ; DNA-Binding Proteins ; genetics ; Humans ; Proteins ; genetics ; Sequence Analysis, DNA ; Two-Hybrid System Techniques ; Yeasts ; genetics