Animals ; Cell Cycle ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; Humans ; Hyperbaric Oxygenation ; K562 Cells ; Mice ; Mice, Nude ; Xenograft Model Antitumor Assays

Objective: To explore and obtain the isolation and culture methods of the highly purified primary rat brain microvascular endothelial cells (BMECs). Methods: The brains of 10 3-week-old Wistar rats were harvested by decapitation, removing the white matter and mincing into approximately 1 mm3. A highly purified brain microvascular fragments were obtained through enzymatic digestion twice, 20% bovine serum albumin (BSA) and 33% continuous Percoll density gradient centrifugation. They were incubated into gelatin-coated 35 mm dish plates and the endothelial medium containing 4 μg/ml puromycins was added. The medium was renewed after 2 days. The endothelial cell growth and its morphology were observed using an inverted microscope. The expression of BMECs markers factor VIII-related antigen (vWF) was detected by immunocytoehemical method and was identified. Results: The endothelial cells grew out around the adherent brain microvascular fragments after culturing for 24 hours, and the cells were spindle-shaped. A fusion was formed after 5 to 6 days. BMECs after the fusion showed a typical "cobblestone-like" appearance. Immunocytochemical analysis showed the vWF expression of BMECs was positive, while the glial fibrillary protein (GFAP) expression was negative. The purity of BMVECs reached more than 96%. Conclusion: This method may successfully obtain the highly purified rat primary BMECs.

Congenital Abnormalities ; Encephalocele ; Humans ; Nose Diseases ; congenital

Animals ; Antineoplastic Agents ; therapeutic use ; Gene Silencing ; Genetic Therapy ; methods ; Humans ; MicroRNAs ; antagonists & inhibitors ; therapeutic use ; Neoplasms ; genetics ; therapy ; RNA Interference ; physiology ; RNA, Small Interfering ; antagonists & inhibitors ; therapeutic use ; RNA-Induced Silencing Complex ; metabolism

To analyze the reasons for the formation of constipation,the causes,clinical manifestations,surgical indications and surgical treatment were analyzed,focusing on slow transit constipation,outlet obstruction constipation and mixed constipation.Grasping surgery indications is stressed in surgical treatment of constipation.Only appropriate procedure can obtain satisfactory results.

ObjectiveTo evaluate the clinical efficiency of 0.1％ tacrolimus ointment combined with paroxetine in patients with neurodermatitis associated with insomnia.MethodsTotally,74 patients suffering from neurodermatitis and insomnia were consecutively enrolled in this study,and randomly assigned into the combination group and control group according to their number(odd or even).Both groups were given topical 0.1％ tacrolimus ointment twice a day,and the combination group was additionally given oral paroxetine 10 mg once a day,for 8 weeks.The outcome parameters included main symptoms of neurodermatitis and Pittsburgh sleep quality index(PSQI) score.ResultsNo significant difference was observed in the cure rate (54.3％(19/35) vs.53.1％ (17/32),P＞ 0.05) or response rate (82.9％ (29/35) vs.81.3％ (26/32),P＞ 0.05),between the combination group and control group.PSQI score was significantly lower in the combination group than in the control group after 8 weeks of treatment(P ＜ 0.05).ConclusionsThe 0.1％ tacrolimus ointment can be used to treat neurodermatitis safely.Antianxiety treatment can reduce the recurrence of neurodermatitis associated with insomnia,and improve the quality of sleep in patients.

Castor(Ricinus communis L.),a high energy-stored and industrial raw-material plant species,is of great value in its application.Progresses in castor biotechnology were summarized in terms of tissue culture and genetic transformation.The results of studies on tissue culture indicated that application of TDZ(thiadiazuron) gave the maximum rate of shoot proliferation in various exogenous hormones when embryo axis was used as explant.On this ground,the in vitro regeneration system was established.The studies on castor genetic transformation showed that the Agrobaterium-mediated method was the most suitable one in different castor transformation methods.The hygromycin is suitable to selection of castor transfomants while castor embryo axis is insensitive to kanamycin.The possibilities of breeding of new castor varieties and improvement of castor production via biotechnology were also discussed.

Objective To investigate the protective effect of topiramate(TPM) on neuronal apoptosis in rats with acute seizures.Methods We treated the PTZ-induced seizure rats with TPM at 80mg/(kg?d)(high-dose group) and 40mg/(kg?d)(middle-dose group) or physiological saline(control group) for 2 weeks.Neuronal apoptosis in CA_1 and CA_3 regions in hippocampus was identified by terminal deoxynucletidyl transferase-mediated dUTP-biotin in situ nick end labeling(TUNEL) assay.Results Two weeks following seizures,TUNEL-positive neurons were detected in CA_1 and CA_3 regions each group.The numbers of TUNEL-positive neurons in CA_1 and CA_3 of control group were(35.83?)4.58 and(36.83?)3.83,(23.50?)2.81 and(25.50?)3.72 of high-dose TPM group,(31.52?)3.43 and(32.35?)4.69 of middle-dose TPM group.There was a very significant difference between high-dose TPM group and control group(all(P)0.05).Conclusion High dose administration of TPM after experimental status epilepticus may attenuate seizure-induced hippocampal neuronal injury.

0.05). Significant difference was observed between Group C+D and Group A+B(P

Traditional Chinese medicine (TCM) emphasizes treatment based on syndrome differentiation, which is the core technology that can embody the characteristics and advantages of TCM diagnosis and treatment. This is based on holistic view to illustrate individual diagnosis and treatment and dynamic treatment. However, RCT, which is frequently used in TCM clinical studies, can not reflect the concept of treatment based on syndrome differentiation. The use of adaptive treatment strategies in clinical researches can adjust therapy in patients according to the changes of diseases, so it can reflect individual diagnosis and treatment and dynamic treatment. From this perspective, the process of treatment based on syndrome differentiation is a process of decision-making of adaptive treatment. So the method for developing adaptive treatment strategies can effectively evaluate the efficacy of different treatment strategies in TCM clinical studies. Adaptive treatment strategies have been used in some clinical studies from some published foreign literatures. However, the use of adaptive treatment strategies in domestic clinical researches is seldom reported. This article discussed the concept of adaptive treatment strategies and their important components in detail. In addition, it presented three methods which can take the delayed effects of the current treatment action into consideration and can compare the overall effect of different adaptive treatment strategies, with a purpose to lay some foundation for future TCM researches in the development of optimal clinical pathway and provide new ideas and methods for researches.