Animals ; Dependovirus ; genetics ; immunology ; Gene Transfer Techniques ; instrumentation ; Genetic Therapy ; instrumentation ; Genetic Vectors ; genetics ; immunology ; Humans

The prescriptions containing pericarpium citri reticulatae that built by Professor. Yan were collected to build a database based on traditional Chinese medicine (TCM) inheritance assist system. After analyzed by data mining, such as apriori algorithm, the frequency of single medicine, the frequency of drug combination, the association rules between drugs and core drug combinations can be get from the database. Through the analysis of 1 027 prescriptions with pericarpium citri reticulatae, these prescriptions were commonly used to treat stomach aches, cough and other syndromes. The most frequency drug combinations were "Citri Reticulatae Pericarpium-Poria", "Paeoniae Radix Rubra-Citri Reticulatae Pericarpium" and so on. The drug association rules that the confidence was 1 were "Glycyrrhizae Radix ex Rhizoma --> Citri Reticulatae Pericarpium", "Paeoniae Alba Radix-Cyperi Rhizoma --> Citri Reticulatae Pericarpium", "Poria --> Citri Reticulatae Pericarpium", and so on. The drugs in the prescriptions containing pericarpium citri reticulatae that built by Professor Yan mostly had the effects of regulating the flow of Qi and invigorate blood circulation, which reflected the clearly thought when making prescriptions.
Citrus ; chemistry ; Data Mining ; Drug Interactions ; Drug Prescriptions ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Practice Patterns, Physicians'

Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.

To clone, express and primarily use human autoantigen Sm D1 in methylotrophic yeast Pichia Pastoris. The gene Sm D1 was cloned by PCR.The PCR product was inserted into the vector pPIC9k. The recombinant plasmid pPIC9k- Sm D1 was transformed into yeast SMD1168 by electroporation. The positive clones were screened in MD plates. The high copy number transformants were rapidly selected by using G418 and were induced by methanol. Supernatants after induction were analyzed by SDS-PAGE and im-munodot. The PCR product was showed about 360 bp in size which was in accordance with predicted. The pPIC9k-Sm D1 showed the same seqencing result with GenBank’s report and restriction enzyme analysis confirmed our prediction. The pPIC9k-Sm D1 positive clone produced an about 16 kD protein which had natural immunogenicity of human autoantigen Sm D1 by SDS-PAGE and immunodot. The sensitivity and specificity of immunodot were 96% and 100%, respectively. The agreement between immunodot and im-munoblot was 98%. Successfully cloning and high-level expression of human autoantigen Sm D1 in methy-lotrophic yeast Pichia pastoris laid a foundation for further research work.

The repair of spinal cord injury is still a complex medicine problem because of its second injuries and involuted pathogenesis. In recent years, spinal cord injury repair by cell-transplantation has showed a big prospect. Although every kind of cell-transplantation was effective, it remained some problem, co-transplantation of cells hit the focus. This paper has reviewed the pathologic change after spinal cord injury, the influential factor and replied strategy for spinal cord regeneration and the advance in co-transplantation of cells for spinal cord injury.

In this study, prescriptions were collected to establish a database based on the traditional Chinese medicine inheritance support system (TCMISS). Such data-mining methods as software's statistical statement module, data analysis module and apriori algorithm were used to analyze the frequency of single drug, the frequency of drug combination, the association rules and the core drug combinations of ostreae concha-containing prescriptions. The results showed that Ostreae Concha-containing prescriptions were frequently used to treat insomnia, vertigo, stomach-ache and other syndromes. The frequently used drugs included Ossis Mastodi Fossilia, Parched Ziziphi Spinosae Semen, Polygoni Multiflori Caulis, Salviae Miltiorrhizae Radix and Paeoniae Rubra Radix. The frequently used drug combinations included "Ostreae Concha and Ossis Mastodi Fossilia", "Ostreae Concha and Parched Ziziphi Spinosae Semen", and "Ostreae Concha and Polygoni Multiflori Caulis". The drug association rules with the confidence coefficient of more than 0. 95 included "Ossis Mastodi Fossilia-->Ostreae Concha", "Cocos Poria-Ossis Mastodi Fossilia-Ostreae Concha", "Ossis Mastodi Fossilia-Parched Ziziphi Spinosae Semen-->Ostreae Concha", and "Salviae Miltiorrhizae Radix-Ossis Mastodi Fossilia--Ostreae Concha".
Animals ; Chemistry, Pharmaceutical ; Data Mining ; Databases, Pharmaceutical ; Drug Prescriptions ; Humans ; Medicine, Chinese Traditional ; methods ; Ostrea ; chemistry ; Physicians ; Software

Based on the data mining methods of association rules and clustering algorithm, the 188 prescriptions for cough that built by Yan Zhenghua were collected and analyzed to get the frequency of drug usage and the relationship between drugs. From which we could conclude the experiences of Yan Zhenghua for the treatment of cough. The results of the analysis were that 20 core combinations were dig out, such as Bambusae Caulis in Taenias-Almond-Sactmarsh Aster. And there were 10 new prescriptions were found out, such as Sactmarsh Aster-Scutellariae Radix-Album Viscum-Bambusae Caulis in Taenian-Eriobotryae Folium. The results of the analysis were proved that Yan Zhenghua was good at curing cough by using the traditional Chinese medicine that can dispel wind and heat from the body, and remove heat from the lung to relieve cough.
Algorithms ; Cough ; drug therapy ; Data Mining ; Drug Prescriptions ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Practice Patterns, Physicians'

The prescriptions including Polygoni Multiflori Caulis that built by Pro. Yan were collected to build a database based on traditional Chinese medicine (TCM) inheritance assist system. The method of association rules with apriori algorithm was used to achieve frequency of single medicine, frequency of drug combinations, association rules between drugs and core drug combinations. The datamining results indicated that in the prescriptions that including Polygoni Multiflori Caulis, the highest frequency used drugs were parched Ziziphi Spinosae Semen, Ostreae Concha, Ossis Mastodi Fossilia, Salviae Miltiorrhizae Radix Et Rhizoma, Paeoniae Rubra Radix, and so on. The most frequent drug combinations were "Polygoni Multiflori Caulis-parched Ziziphi Spinosae Semen", "Ostreae Concha-Polygoni Multiflori Caulis", and "Polygoni Multiflori Caulis-Ossis Mastodi Fossilia". The drug association rules of confidence coefficient 1 were "Ostreae Concha-->Polygoni Multiflori Caulis", "Poria-->Polygoni Multiflori Caulis", "parched Ziziphi Spinosae Semen-->Polygoni Multiflori Caulis", and "Paeoniae Alba Radix-->Polygoni Multiflori Caulis". The core drug combinations in the treatment of insomnia were Ossis Mastodi Fossilia, Polygoni Multiflori Caulis, Salviae Miltiorrhizae Radix et Rhizoma, Ostreae Concha, Polygalae Radix, Margaritifera Concha, Poria, and parched Ziziphi Spinosae Semen. And the core drug combinations in the treatment of obstruction of Qi in chest were Salviae Miltiorrhizae Radix Et Rhizoma, Polygoni Multiflori Caulis, parched Ziziphi Spinosae Semen, Trichosanthis Fructus, Allii Macrostemonis Bulbus, and Paeoniae Rubra Radix.
Data Mining ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional ; methods ; Plant Stems ; chemistry ; Polygonaceae ; chemistry ; Prescriptions

Objective To analyse the effects of serum estradiol levels during controlled ovarian hyperstimulation (COH) on the outcomes of in vitro fertilization and embryo transfer (IVF-ET). Methods The clinical data of 472 patients undergoing IVF-ET with GnRH analogues recombinant FSH long protocol were retrospectively analysed. The area under the curve (AUC) of estradiol (E2) level was calculated during COH, and patients were categorized into groups according to the percentile of AUC of E2(AUCE2) during COH. The general characteristics and parameters related to the outcomes of IVF-ET were compared among groups. Results The 10th percentile and 90th percentile of AUCE2 were 3 347.0 pmoL/L and 14 414.3 pmol/L, respectively. Four hundred and seventy-two patients were divided into lower reaction group (AUCE2 3 347.0 pmol/L, n=48), normal reaction group (14 414.3 pmol/L>AUCE2 > 3 347.0 pmol/L, n=376) and higher reaction group (AUCE2≥14 413.3 pmol/L, n=48). There was no significant difference in age, body mass index, baseline follicle stimulating hormone level, time of treatment with gonadotropin, endometrium thickness on day of transfer and embryos transferred(P>0.05). Compared with lower reaction group and normal reaction group, the number of oocytes per retrieval and number of embryos frozen were significantly larger(P<0.01) and the mild/severe ovarian hyperstimulation syndrome rate was significantly higher in higher reaction group(P<0.05). There was no significant difference in fertilization rate, cumulative embryo score, high-grade embryo rate, clinical pregnancy rate and implantation rate among groups (P> 0.05). Conclusion Sustained snpraphysiological serum E2 levels during the COH process do not adversely affect the quality of oocytes and embryos, clinical pregnancy rate and implantation rate to some extent in IVF-ET.

AIM:To investigate the effect of RWDD3 gene silencing on the biological characteristics of human glioma U251 cells.METHODS: A lentiviral vector expressing RWDD3 shRNA was constructed and transfeeted into the U251 cells.The expression of RWDD3 at mRNA and protein levels was detected by real-time PCR and Western blot , re-spectively .The cell activity was determined by MTT assay .The colony formation ability was detected by the colony forma-tion assay .The cell proliferation ability was detected by BrdU incorporation assay .The cell invasion and migration were evaluated by Transwell assay .Flow cytometry was used to monitor the changes of cell cycle distribution and apoptosis .RE-SULTS:Recombinant lentivirus was successfully transfected into U 251 cells.Compared with the cells transfected with the scrambled shRNA and control cells, the cell activity, colony formation ability, and the invasive and migratory activities were inhibited, the cell cycle was arrested in G 0/G1 phase, and the apoptosis was increased in the U 251 cells transfected with RWDD3 shRNA ( P <0.05 ) .CONCLUSION: RWDD3 plays a vital role in proliferation and invasion of glioma cells.It may serve as a potential target of gene therapy for glioma .