Objective: To investigate the possible effects of Pancreatic duodenal homeobox-1((Pdx-1))expression in transdifferentiation of bone marrow mesenchymal stem cells(MSC) in vitro. Methods: The eukaryotic expression vector containing Pdx-1 was constructed.After such vector was transfected into MSC using Superfect,G418 was used to select the positive cells.Then both(Pdx-1~(+)MSC)and((Pdx-1~-MSC)) were induced to transdifferentiate in vitro.The expressions of insulin were analyzed by immunohistochemistry staining. Results: Restricted enzyme analysis and sequencing showed that interest gene segment was consistent with that in Gen Bank,Recombination vector was effectively transformed into MSC demonstrated by fluorescence microscope;insulin-producing cells from Pdx-1~(+)MSC were higher than that from Pdx-1~(-)MSC[(28.23?2.56)% and(7.08?2.69)%,respectively]. Conclusion:Pdx-1 can promote adult rat MSC to transdifferentiate into insulin-producing cells in vitro,and this approach might lead to a widespread cell replacement therapy for type I diabetes.

Many previous studies have shown that auditory cortical neurons are sensitive to sound spatial information, however, the mechanism of sound spatial coding is still not fully understood. Until now, detail studies on sound spatial coding have not been reported in the rat primary auditory cortex. Using electrophysiological technique, spatial response areas of 151 neurons in the rat primary auditory cortex were investigated. The relationships between spike counts and average first-spike latencies in the spatial response areas were analyzed. The results showed that, the majority (52.32%) of cortical neurons exhibited contralateral preference in the frontal auditory space whereas other neurons exhibited ipsilateral preference (18.54%) and midline preference (18.54%), and only a few neurons were included in the category of omnidirection (3.31%) and complex (7.28%). For the majority of cortical neurons, the arithmetic center of the preferred spatial area were distributed in the up and middle portion of the contralateral space relative to the recording side. Most neurons responded strongly to stimuli from their preferred space with shorter average first-spike latencies, and responded weakly to stimuli from non-preferred space with longer average latencies. In the spatial response area, the spike counts were negatively correlated with average first-spike latencies. The auditory cortex might use the information of both spike counts and average response time to code sound spatial information.

Purpose:To investigate expression of keratinocyte growth factor(KGF) messenger ribonuclunc acid in human NSCLC and to study the role of KGF in the development of NSCLC. Methods:The expression of KGF mRNA in 50 cases with NSCLC was detected by in situ hybridization (ISH). Results:On ISH slides, positive KGF mRNA was mainly shown as fusiform stain in plasma of fibroblast and blood vessel smooth muscle cell in mesenchymal of NSCLC, also, some parenchyma cell plasma was stained. The positive rate of KGF mRNA in tumor(86%) is statistically higher than that in normal tissue(24%)(P

Objective To explore the clinical feature and treatment method of the dopa responsive dystonia (DRD).Methods To observe the clinical manifestation,auxiliary test and the response on the treatment of levodopa in 6 patients from 4 families.Results 1 patient for childhood onset and 2 patients for adolescent onset with DRD, first symptom showed leg dystonia and toe walking with difficulty, whereas 3 patients of adult onset showed tremor and rigidity. Babinski sign was presented in 3 cases. Diurnal fluctuation in symptom severity occurred in all cases. All patients had obvious response to small dose levodopa in 1～6 days. In 2 cases, the effect was maintained for 7 years, without increasing the intake of levodopa.Conclusion The clinical feature of DRD was more remarkable,the therapeutic effectiveness of levodopa was rapid,sustained and obvious.

Objective: To establish the quality standards for Lingyangqingfei capsules. Methods: Microscopic identification of products, TLC identifications of Fructus Gardeniae and Cortex Moutan were carried out.Baicalin was determined by HPLC. Results: Baicalin in Lingyangqingfei capsules was about 1.5mg/capsule. The average recovery was 99.82% and RSD was 0.96%. Conclusion: The result is accurate and the reproducibility is good. This method can be used as the quality control of this preparation.

AIM:To construct a eukaryotic expression vector containing pancreatic duodenal homebox-1(PDX-1)and to elevate the expression efficiency of exogenous gene in rat bone marrow mesenchymal stem cells(MSCs).METHODS:Recombinant vector containing PDX-1 was constructed.Flow cytometry was used to identify the cell cycle of bone marrow mesenchymal stem cells(MSCs)cultured in vitro.Recombinant vector containing PDX-1 was transfected into bone marrow MSCs using superfect in medium.After being selected by G418,RT-PCR and Western blotting were used to investigate the expression of PDX-1 in MSCs.RESULTS:Restricted enzyme analysis and sequencing showed that PDX-1 gene segment was consistent with that in GenBank.Flow cytometry showed that there were about 85.9% cells at the cell cycle of G_0/G_1.The whole cells transfected emitted green fluorescence under flow cytometry.The efficiency of transfection was above 40%.RT-PCR and Western blotting demonstrated that there was expression of PDX-1 in transfected bone marrow MSCs.CONCLUSION:Recombinant vector containing PDX-1 was constructed successfully.Superfect mediated expression of exogenous gene in bone marrow MSCs in a high efficiency,and bone marrow MSCs containing exogenous gene are an ideal cells for gene therapy.

The widespread application of traditional medicine has attracted increasing attention. To promote the security of traditional medicine, WHO has established a series of global policies and strategies with consistency since 1960s. It also formulated guidelines for harvesting, production and operation, and quality control of botanical medicine according to the development of traditional medicine in different times. Thanks to the technical support of WHO, member states have established technical regulations for traditional medicine according to their own situations, and made significant achievements in the promotion of traditional Chinese security.

Objective To investigate the effect of Notch signaling during bone marrow mesenchymal stem cells (BMSC) differentiating into islet in vitro.Methods The specific inhibitor of γ-secretase DAPT was used to inhibit the Notch signaling pathway.Mter induction,DTZ staining,indirect immunofluorescence staining,RT-PCR and Westenn blotting were used to detect the expression of insulin,glucagon,Pdx-1 and Ngn3.Results (1) Identification of BMSCs:Indirect immunofluorescence staining showed that BMSCs could express CD59 and CD90,which both were makrers of mesenchymal stem cells.Besides,BMSCs could express nerve culluar markers such as NSE,GFAP,suggesting multi-directional differentiation.(2) The result of MTT showed DAPT could inhibit the cell proliferation in a time-dependent manner and a dose-dependent mannar.Besides,DAPT could inhibit the expression of target gene of Notch signal pathway in a time-dependent manner and a dosedependent mannar.After treated by 1,5,20 μmol DAPT,the expression of Hes1 had reached to 92.06％,71.40％ and 46.89％ of controls respectively,suggesting efficiency of inhibition on Notch reached 7.94％,28.6％ and 53.11％ respectively (all P＜ 0.05).(3) Indirect immunofluorescence staining showed the expression of pancreas-specific markers such as insulin and glucagon were much higher in DAPT treated BMSCs than that in controls,which was confirmed by RT-PCR and Western blotting analyses.The proportion of insulin-producing cells differentiated from DAPT treated BMSCs was (74.03 ± 3.96)％,which was higher than that from controls[(36.49 ± 3.24)％,P＜ 0.05].(4)Furthermore,RT-PCR and Western blotting analysis showed that the expressions of Pdx-1 and Ngn3 were earlier than that of insulin and glucagon,and the expressions of Pdx-1 and Ngn3 were higher in DAPT treated BMSCs than that in controls.Conclusions Notch signaling pathway plays a role in the differentiation of BMSCs into islet in vitro.Pharmacological interference with Notch signaling pathway may provide a novel method to obtain islet for therapeutic use.

Chinese medicine has developed rapidly in the world, but the international standard system has not been established, further affecting the healthy development of Chinese medicine. Technical status of the member states are closely related to the current legislation and regulations in the development of Chinese medicine, which is important for developing international standards for TCM strategies. By the findings of ISO Technical Committee of Traditional Chinese Medicine (ISO/TC249) 7 European P member (Participating countries) International Traditional/Alternative Medicine (TM/CAM) legislative status, herbal medicine in each member country has a large market, a good environment for the development of some countries. After“The EU Traditional Herbal Medicinal Products Directive 2004/24/EC”issued by the member states for the registration and management in European, regulation of herbal products become more stringent. There are relatively small quantity of TM/CAM practitioners, lack of education for practitioners and standardized management.