Dao-di Herbs is specificity and locality, and its unique phenotypic features is closely related to the growth and development of medicinal plants. In addition to traditional genetic, epigenetic play an important role in formation of Dao-di herbs. This paper introduces the concept of epigenetic and the role of DNA methylation in the gene expression regulation. We further prospects epigenetic mechanism in study of Dao-di herbs formation from specific phenotype and regional analysis. And study on the relationship of epigenetic and Dao-di herbs will provide a basis for quality assessment and identification of Chinese drugs.
DNA Methylation ; Drugs, Chinese Herbal ; standards ; Epigenesis, Genetic ; Gene Expression Regulation, Plant ; Plants, Medicinal ; genetics

Apoptosis is a spontaneous process of cell-suicide process triggered in response to physiological and pathological stress stimul, which is to regulate the developmental of body, control cell senescence, maintain a stable internal enviroment in multicellular organisms. The initial and progress of apoptosis is precisely controlled, which is a unique and complex signal system. In this review, it introduced the relationship between apoptosis and bcl-2 gene family, C-myc, tumor suppressor gene p53, Caspase protease family and Fas, and Summaries the detection methods.

In the research field of quality control in Chinese medicinal materials, variation in active ingredients of medicinal plant is always the key and hot issues. With the development of high-throughput sequencing technologies and reducing cost, a large numbers of genes from medicinal plant were cloning and provide a solid foundation for further research of gene structure and its biological function, and also provides conditions for explore active ingredient variation and its quality control from the perspective of molecular pharmacognosy. This paper introduces the concept of homologous gene, gene duplication and classification. We prospect the function of duplicated genes in the role of molecular mechanism research about variation in active ingredients, aiming at providing a new way for medicinal materials quality control.
Drugs, Chinese Herbal ; analysis ; Gene Duplication ; Plant Proteins ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Quality Control

Objective To construct recombinant full length genotype B and C hepatitis B virus (HBV)and to examine HBV DNA replication and hepatitis B surface antigen(HBsAg),hepatitis B e antigen(HBeAg)expressions in Huh7 cells. Methods The full length genotype B and C HBV DNA were extracted and amplified from two HBV infected patients. The recombinant plasmids were constructed by inserting the amplified HBV fragments into the eukaryotic expression vector,pHY106,which were then transfected into Huh7 cells. The cells transfected with blank pHY106 vector were used as control. HBV DNA replication at 72 hours of transfection was detected by Southern blot. The HBV DNA levels in Huh7 cells at 24,48,72,96 and 120 hours of transfection were determined by real-time polymerase chain reaction(PCR).Meanwhile, the HBsAg and HBeAg expression levels in the supernatants at 24,48,72,96 and 120 hours were determined by enzyme linked immunosorbent assay(ELISA).Results The recombinant plasmids expressing genotype B or C HBV DNA were successfully constructed.The HBV replicative intermediates in HBV core particles,including rcDNA dsDNA and ssDNA,were detected by Southern blot.HBV DNA level could reach 8 lg copy/mL which was by real-time PCR. HBsAg and HBeAg levels determined by ELISA peaked at 72 hours after transfection and then declined gradually. Conclusions The recombinant plasmids inserted with genotype B or C HBV DNA are constructed successfully, which can express high levels of HBsAg and HBeAg in Huh7 cells. This system provides a platform for studying the pathogenesis of B and C genotype HBV, the interaction between HBV and host, as well as exploiting new drugs against HBV.

BACKGROUND: Magnesium alloy studies in orthopedic field have been carried out,and good biocompatibility has been reported.However magnesium alloys have not yet been researched in the intestine.OBJECTIVE: The biodegradable magnesium-zinc alloy samples are implanted around the rat cecum to investigate the biocompatibility in rat.METHODS: Sprague Dawley rats were randomly divided into magnesium alloy group,medical titanium group and the sham-operated group.Then magnesium-zinc alloy samples with the dimension of 5 mm × 1 mm× 1 mm were embedded in the cecum incision in the magnesium alloy group.The medical titanium was embedded in the medical titanium group,and just suture in the sham-operated group.Prior to surgery and at 7,14,21 and 28 days after operation,the serum glutamic-oxaloacetic transaminase,creatinine and magnesium ion concentration were examined in each group.X-ray film on implanted region.The pathological changes in liver,kidney and cecum were examined.RESULTS AND CONCLUSION: There were no significant differences in serum glutamic-oxaloacetic transaminase,creatinine and magnesium ion concentrations among each group(P > 0.05).Magnesium-zinc alloy degraded gradually during 28 days.The pathology of liver,kidney and cecum was normal.Results suggested that magnesium-zinc alloy had no obvious effect on the cecum.The degradation time to play a fixed function of time was longer than the intestinal healing time,with good biocompatibility.Magnesium-zinc alloy can be used as anastomotic nail for stomach intestine anastomat.

High resolution melting (HRM), an important technology for genotyping and mutation scanning, has broad prospects in the authentification of traditional Chinese medicine. This paper selected universal trnH-psbA primers and used HRM to establish a new methods for identification of Akebia herbs. PCR was conduct at the annealing temperature of 58 degrees C and 35 cycles. The range of the DNA template concentration, the primer concentration and the Mg2+ ion concentration were further analyzed. The results showed the Tm values of Caulis Akebiae was (81.84 ± 0.16), (85.28 ± 0.16) degrees C and Caulis Clematidis Armandii was (83.22 ± 0.19) degrees C and Caulis Aristolochiae manshuriensis was (81.67 ± 0.14) degrees C, (84.24 ± 0.10) degrees C with 5-125 mg - L-' DNA template, 0.4 μmol x L(-1) primer, 2.0 mmol x L(-1) Mg2+. This method can achieve the authentification of Akebia herbs and is simple, fast, high-throughput, visual.
Chemistry Techniques, Analytical ; methods ; DNA, Plant ; chemistry ; genetics ; Genotype ; Magnoliopsida ; chemistry ; classification ; genetics ; Phylogeny ; Transition Temperature

Molecular identification of Chinese traditional medicine has come from laboratory research into application, but there are some misunderstandings and problems emerging after rapid development. In this paper, we discuss the usage principle, hot field and technology innovation in molecular identification of Chinese traditional medicine. And molecular identification of traditional Chinese medicine has scientific and objective basis, follows the certain systematic research background, and adopts practical principles to establish case by case multi-class identification system. In order to achieve rapid, on-site, high throughput, low cost of traditional Chinese medicine identification purpose, molecular identification technology is further developing for meet the actual needs and the laboratory results further transformation in the service of traditional Chinese medicine industry.
China ; Drugs, Chinese Herbal ; chemistry ; standards ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; classification ; genetics ; Quality Control

Objectives To screen the differentially expressed proteins in serum of population chronically exposed to arsenic in drinking water,thus to provide candidate protein biomarkers for arsenic exposure and arsenicosis.Methods Subjects were selected from the drinking water type of endemic arsenicosis areas in Shanxi province,China.Demographic characteristics,history of arsenic exposure,cigarette smoking,alcohol drinking,health and other information were collected using questionnaire.The subjects were divided into low-arsenic group (with arsenic in drinking water ＜ 10 μg/L),medium-arsenic group( 10 - 50 μg/L),high-arsenic group( ＞ 50 μg/L),and arsenicosis group(the drinking water with arsenic ＞ 50 μg/L was replaced by low arsenic water ＜ 10 μg/L).The number of cases in each group was 30.The arsenicosis patients were diagnosed according to “Standard of Diagnosis for Endemic Arsenism” (WS/T 211-2001 ).With the principle of informed consent,blood samples were collected.Differentially expressed serum proteins of different arsenic exposure groups and arsenicosis group were screened by two-dimensional differential gel electrophoresis(2-D DIGE),and further identified by mass spectrometry (MS).Results An average of (1299 ± 167) protein spots were identified in 6 gel images and 688 protein spots were discovered repeatedly in at least 5 gels.There were 33 protein spots differentially expressed among low-,medium- and high-arsenic groups P ＜ 0.01).Fifty four protein spots were significantly different among low-,medium-,high-arsenic exposure groups and arsenicosis group(P ＜ 0.01 ).Twenty five protein spots were selected for MS analysis,and13 protein spots were identified.Compared with low-arsenic group,the expressions of apolipoprotein A-Ⅳ,retinol binding protein,and estrogen receptor hypothalamic isoform in medium- and higharsenic exposure groups were down regulated,and the expressions of component 4A and 4B were up regulated.Compared with low-,medium- and high-arsenic groups,the expressions of beta-2-glycoprotein Ⅰ,Keratin 1,hemopexin,complement C1r subcomponent,and ficolin-3 in arsenicosis group were down regulated,and the expressions of pigment epithelial-differentiating factor,alpha-1-microglobulin and carboxypeptidase N catalytic chain were up regulated.Conclusions Chronic arsenic exposure can significantly change population's serum protein expression.Differentially expressed proteins in arsenicosis patients will not decline with the decline of arsenic in a short term.Whether or not the differentially expressed proteins identified in this study can be used as biomarkers for arsenic exposure and arsenicosis needs to be further verified.

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, with the method of reverse transcription polymerase chain reaction (RT-PCR), this study cloned full-length cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase gene from Salvia miltiorrhiza bge.f.alba, this sequence is named as SmHDS and its GenBank registration number is KJ746807. SmHDS, 2 529 bp long, contains an ORF of 2 229 bp, encodes 742 amino acids, including 5' UTR 170 bp and 3' UTR 130 bp. Using bioinformatics software, having made a homology analysis of the obtained sequence, we can have a conclusion that SmHDS have a close genetic relationship with HDS of Salvia miltiorrhiza. Analysis result of prokaryotic expression revealed that in Escherichia coli, SmHDS expressed target proteins which in size are comparable with the protein predicted. Meanwhile, the 4 factors which can influence the protein expression were optimized, the 4 factors are inducing temperature, inducing time, IPTG concentrations and density of inducing host bacterium (A600). The optimal expression conditions of SmHDS were 30 degrees C until the A600 is 0.6, and add IPTG to a final concentration of 0.2 mmol x L(-1), and the induction time of 20 h. It provides theoretical basis for the further study of the function of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase in the biosynthesis of tanshinone compounds.
Cloning, Molecular ; DNA, Complementary ; genetics ; Diterpenes, Abietane ; biosynthesis ; Enzymes ; biosynthesis ; genetics ; Escherichia coli ; metabolism ; Plant Proteins ; biosynthesis ; genetics ; Salvia miltiorrhiza ; enzymology ; genetics

ObjectiveTo investigate the effect of total disc replacement (TDR) in the treatment of cervical spondylotic myelopathy(CSM) with sympathetic symptoms.MethodsTwenty-seven patients (15 males and 12 females) with CSM at single level accompanied by sympathetic symptoms,who underwent TDR surgery(Prestige or Discover prosthesis) with the posterior longitudinal ligament (PLL) resected at the area of decompression between October 2008 and May 2011,were retrospectively analyzed.All patients were followed up for at least 6 months (average,15.1 months).Clinical and radiologic evaluations were obtained preoperatively,1 week postoperatively,and at the final follow-up.The sympathetic symptoms were scored by our original 20-point system.The clinical outcomes were assessed by Japanese Orthopedic Association (JOA) scoring system and the Short Form-36 Health Survey (SF-36).ResultsThe sympathetic symptoms were improved in all patients and the score was significantly improved after surgery.The sympathetic symptoms scores were (8.5±2.5) points preoperatively,(1.6±1.4) points 1 week postoperatively,and (2.3±1.1) points at the final follow-up.The patient's subjective satisfaction was excellent in 15 patients,good in 8,fair in 4,with an excellent and good rate of 85.2％.The corresponding JOA scores were(10.8±2.4) poiuts, (11.1±2.5) points,and (14.9±1.4) points,respectively.The SF-36 scores showed statistical improvements from preoperative (102.7±8.7) points to postoperative (129.8±5.5) points.Based on X-ray examination,the range of motion of the treated segment were reserved.During the follow-up period,there was no prosthesis subsidence or excursion.ConclusionThe CSM patients with sympathetic symptoms could be managed successfully with TDR.And thoroughly resection of the PLL may be the key factor for good prognosis.