Edema, as one of common clinical diseases, could be treated by taking medicines and adopting external therapies with traditional Chinese medicines (TCM). In recent years, there have been many clinical and basic studies concerning external therapies with TCM on edema Data showed that the external therapies are mostly composed of such purgating drugs as Rhei Radix et Rhizoma, Natrii Sulfas and Pharbitidis Semen, heat-clearing drug such as Phellodendri Chinensis Cortex and resuscitation-inducing drug such as Borneolum Syntheticum. The study showed that ingredients of external therapies did not pass through hilum and hepatic system, and thus avoided the first pass effect of livers. They enabled effective components of drugs to be rapidly absorbed through pores and skins, strengthened the effect against edema, shortened the treatment course, decreased side effects, and were convenient and inexpensive. External therapies with TCM could play unique advantages in inhibiting edema in the future clinical studies.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Edema ; drug therapy ; Humans

Objective To clone a new human gene 5 trans-activated by pre-S1 protein of hepatitis B virus (HBV), PS1TP5, and explore its structure and function by bioinformatics analysis. Methods PS1TP5 was amplified by reverse transcription-polymerase chain reaction (RT-PCR) technique by using HepG2 cDNA as template and inserted into pGEM-T vector by TA cloning. Recombinant eukaryotic expression vector pcDNA TM 3.1/myc-His A-PS1TP5 had been constructed by subcloning, followed by restriction enzyme digestion analysis and sequencing. Bioinformatic methods were used to analyze its possible physical and chemical characters, structure, and function. Results PS1TP5 was successfully amplified and cloned into pGEM-T and pcDNA TM 3.1/myc-His A vector by RT-PCR from HepG2 cDNA. The new gene had been confirmed by sequencing after PCR identification and restriction enzyme digestion and named as PS1TP5 because of its trans-active function. The sequence for the PS1TP5 gene had been deposited into GenBank, the accession number was AY427953. Bioinformatics analysis showed that its ORF was 438bp and translated a protein of 145 aa. Conclusion A new gene-PS1TP5 has been recognized, and its recombinant eukaryotic expression vector (pcDNA TM 3.1/myc-His A-PS1TP5) has been constructed. These results will certainly bring some new clues for the study of the biological function of new gene and pathogenesis of chronic hepatitis B.

Objective To clone and identify human genes transactivated by homo sapiens HCV FTP2 by constructing a cDNA subtractive library with suppression subtractive hybridization tech- nique.Methods Suppression subtractive hybridization(SSH)and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV FTP2.The mRNA was iso- lated from HepG2 cells transfected pcDNA3.1(-)-HCV FTP2 and pcDNA3.1(-)empty vector re- spectively,and SSH method was employed to analyze the differentially expressed DNA sequence be- tween the two groups.After digestion with restriction enzyme Rsa I,small-size cDNAs were ob- tained.Then tester cDNA was divided into two groups and ligated to the specific adaptor I and adap- tor 2 respectively.After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR and then was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain DH5?.Futhermore,the cDNA was se- quenced and analyzed in GenBank with Blast search after PCR.Results The subtractive library of genes transactivated by HCV FTP2 was constructed successfully.The amplified library contains 71 positive clones.Colony PCR shows that 56 clones contain 200～1000 hp inserts.Sequence analysis was performed in 24 clones randomly,and the full length sequences were obtained with bioinformatics method.Altogether 20 coding sequences in total were obtained,consisting of 19 known and 1 un- known.Conclusion The obtained sequences may be target genes transactivated by HCV FTP2,and some genes coding proteins involved in cell cycle regulation,metabolism and cell apoptosis.

Objective To study the exact function of HBeBP4A so as to investigate the gene expression of HBeBP4A in yeast cell.Methods Reverse transcription polymerase chain reaction(RT-PCR)was employed to amplify the gene of HBeBP4A from recombinant plasmids pcDNA 3.1/myc-HisA-HBeBP4A,and the gene was cloned into pGEM-T vector.The gene of HBeBP4A was cut from pGEM-T-HBeBP4A vector and cloned into yeast expressive plasmid pGBKT7,and pGBKT7-HBeBP4A was then transformed into yeast AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and Western blotting hybridization.Results HBeBP4A gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The SDS-PAGE and Western blotting assay showed that the relative molecular weight of the expressed product was about 61.37kD,and HBeBP4A protein existed in yeast cells.Conclusion The findings suggested that HBeBP4A was successfully expressed into yeast system.

Objective To investigate the activity of HBV pre-S2 protein on thioredoxin reductase 1 (TXNRD1) gene promoter. Methods TXNRD1 gene promoter DNA sequence was identified in GenBank by bioinformatics and amplified from HepG2 genome by polymerase chain reaction (PCR). The amplified product was cloned into pCAT3-Basic reporter vector,named pCAT3-TXNRD1p. The HepG2 cells were transfected by pCAT3-TXNRD1p,and then co-tranfected by pCAT3-TXNRD1p and pcDNA3.1(-)-preS2 plasmids. The choloraphenical acetyltransferase(CAT)activity was assessed by enzyme linked immunosorbent assay(ELISA). Results The results indicate that HepG2 cells transfected by pCAT3-TXNRD1p had higher activity of CAT than that transfected by pCAT3-Basic. The expression of CAT in HepG2 cells co-transfected by pCAT3-TXNRD1p and pcDNA3.1(-)-preS2 was 2.2 times higher than that with pCAT3-TXNRD1p. Conclusions The TXNRD1 gene promoter identified in this study has transcription activity and HBV pre-S2 protein can transactivate the expression of TXNRD1 gene.

Objective To screen promoter binding protein of cyclin B2 by using human liver cDNA library, and investigate the expression and regulation mechanism of cyclin B2 gene. Methods By using cyclin B2 biotinylated promoter DNA as the selective molecule, the T7select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment, and the DNA fragment was cloned into pGEM-Teasy vector. Results Sequence analysis was performed in 20 positive plaques, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 6 coding sequences were obtained, all of which were known ones. Conclusion Cyclin B2 promoter binding proteins were screened. The results will be useful for further study the expression and regulation mechanism of cyclin B2.

The secondary development of traditional Chinese medicines (TCMs) is an important content of TCM modernization process, as well as an important path for developing new TCM drugs. Under the guidance of the system theory, in response to the lack of the overall guideline and practical methods for the secondary development of TCMs at present, we introduced the overall thought of the secondary development of major TCM varieties, as well as the roles and contents of clinical research, pharmacology and pharmaceutics in the process of the secondary development of major TCM varieties, so as to provide systematic strategies and methods for the development of major TCM varieties.
Biomedical Research ; Drug Discovery ; methods ; Humans ; Medicine, Chinese Traditional ; methods ; Pharmacology

The secondary development of major traditional Chinese medicine varieties is one of important links during the modernization, scientification and standardization of traditional Chinese medicines. How to accurately and effectively identify the pharmacodynamic material basis of original formulae becomes the primary problem in the secondary development, as well as the bottleneck in the modernization development of traditional Chinese medicines. On the basis of the existing experimental methods, and according to the study thought that the multi-component and complex effects of traditional Chinese medicine components need to combine multi-disciplinary methods and technologies, we propose the study thought of the material basis of secondary development of major traditional Chinese medicine varieties based on the combination of in vivo and in vitro experiments. It is believed that studies on material basis needs three links, namely identification, screening and verification, and in vivo and in vitro study method corresponding to each link is mutually complemented and verified. Finally, the accurate and reliable material basis is selected. This thought provides reference for the secondary development of major traditional Chinese medicine varieties and studies on compound material basis.
Absorption ; Animals ; Drug Discovery ; methods ; Humans ; Medicine, Chinese Traditional ; methods

Acinetobacter Infections ; complications ; diagnosis ; therapy ; Acinetobacter baumannii ; Animals ; Birds ; Humans ; Influenza A Virus, H7N9 Subtype ; Influenza, Human ; complications ; diagnosis ; microbiology ; therapy

Objective To evaluate the clinical efficiency of intermittent photic stimulation (IPS) with pattern and red color plastic plates in video-electroencephalography(EEG).Methods Three hundred and fifty-eight patients hospitalized in Wuhan Children's Hospital from March 2013 to March 2014 were selected,and they were examined by using stroboscope with a red plastic plate,a dots printed plastic plate,and ordinary white flicker for photic driving response (PDR),photoparoxysmal responses (PPR),photoconvulsive response (PCR).The results of patients with PDR,PPR,PCR were analyzed.Results The PDR synchronously evoked by the three flickering patterns were in 29 cases,and the amplitude of photic drivings evoked by ordinary white flicker and red flicker were higher than that of flickering dot pattern [(30.294 ± 7.767) μV,(31.103 ± 8.920) μV vs (24.436 ± 8.075) μV],and there were significant differences(t =2.983,2.815,P =0.003,0.008).However,there was no significant difference between ordinary white flicker and red flicker (t =0.368,P =0.710).The PPR evoked were in 17 cases,and 16 cases (94.12％) of them were evoked by red color,while 10 cases (58.82％) wcre evoked by white light,and 1 case only evoked by flickering dot pattern.However,stronger intensity and longer duration time of PPR were evoked by red color than by white light.In 3 patients with positive PCR,the epileptic seizure was more likely to be evoked by red color or pattern plastic plate.Conclusions IPS with red and pattem plastic plate is more potent in eliciting photosensitive epilepsy in video-EEG than the ordinary white light.Moreover,it could reduce the discomfort of eyes without influencing PDR in comparison to the ordinary white light.