The morphological indexes of the Scuophularia ningpoensis seedlings including the longth, diameter and weight were measured, clustering analysis was used to set up the standard quality grading of seedlings of S. Ningpoensis by SPSS. Field experiment was carried out to measure the indicators of plants growth and development, the yield and the quality. The results showed that the growth and yield of class I seedlings were better than those of class II and III. The content of main active ingredients was affected barely by seedlings classification. To ensure the quality, class II seedlings or above should be used for plantation. The established quality classification standard of S. ningpoensis seedlings was scientific and feasible, and provides the basis for the standardized cultivation of S. ningpoensis.
Drugs, Chinese Herbal ; analysis ; standards ; Quality Control ; Scrophularia ; chemistry ; classification ; growth & development ; Seedlings ; chemistry ; growth & development

BACKGROUND:Astragaloside and tanshinone IIa are the effective components of traditional Chinese medicine treatment of myocardial ischemia, and its role in bone marrow mesenchymal stem cel s differentiation into myocardium-like cel s remains unclear.
OBJECTIVE:To investigate the effect of tanshinone IIa and astragaloside on the differentiation of bone marrow mesenchymal stem cel s into myocardium-like cel s.
METHODS:The maximal non-toxic concentrations of tanshinone IIa and astragaloside were measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, to define the dose of the two in the induced differentiation of bone marrow mesenchymal stem cel s into myocardium-like cel s. The isolated and purified bone marrow mesenchymal stem cel s were divided into five groups:astragaloside group, tanshinone IIa group, astragaloside+tanshinone IIa group, 5-azacitidine group, and blank control group. The expression of gap junction connexin 43 and troponin was determined with enzyme-linked immunosorbent assay.
RESULTS AND CONCLUSION:The expression of gap junction connexin 43 and troponin in astragaloside group, tanshinone IIa group, astragaloside+tanshinone IIa group, 5-azacitidine group was higher than that in blank control group (P<0.01). The astragaloside+tanshinone IIa group showed a higher expression of gap junction connexin 43 and troponin than astragaloside group and tanshinone IIa group (P<0.01). There was no significant difference in the expression of gap junction connexin 43 and troponin between astragaloside+tanshinone IIa group and 5-azacitidine group (P>0.05). A combined use of astragaloside and tanshinone IIa can induce bone marrow mesenchymal stem cel s to differentiate into myocardium-like cel s, and their joint role is better than the role of a single ingredient.

To establish a cell model in vitro that stable expressing HBV by integrating HBA1.3 DNA into cell chromosome. The HBV1.3 full-length DNA was obtained by digested pGEM-HBV1.3 plasmid with HindIII and then was linked with PU-21 vector digested by HindIII. This was resulted in generation of a recombined plasmid named PU21-HBV plasmid. The recombined plasmid was introduced into HepG2 cells by electroporation. The transfected cells were screened with G418. The insertion and expression of HBV were identified by X-gal staining, RT-PCR and Southern blot. The result of PU21-HBV plasmid sequence demonstrated that HBV1.3 DNA was linked correctly with PU-21 vector. A series of positive cell colonies were obtained with G418 screening followed transfecting PU21-HBV plasmid into HepG2 cells. The results of Southern blot and RT-PCR exhibit that HBV1.3 DNA had successfully integrated into the chromosomes of HepG2 cells and had functional HBV gene transcription. HBV1.3 DNA was inserted into HepG2 genome and could stable transcript HBV RNA. The stable HBV expression cell line was constructed successfully. There are LoxP sites in the trapping vector PU21. With the Cre enzyme, interesting genes could be excganged into the LoxP sites. Therefore, double stable expression of interesting gene and HBV cell lines could be generated. The cell lines will be useful for further research some target gene function on replication of HBV.

OBJECTIVETo investigate the relationship between efficacy of Chaihu Taoren Decoction (CTD) and the polymorphism of valine-alanine missense mutation of 9th codan (Val9Ala, T1183C) of superoxide dismutase (SOD) in patients with tardive dyskinesia (TD).

METHODSSeverity of TD was assessed by abnormal involuntary movement scale (AIMS), and the psychologic symptoms were rated by the positive and negative symptoms scale (PANSS). The sample size consisted of 119 patients with TD assigned to the TD group, 129 patients of chronic schizophrenia with the general condition matched strictly with that of the enrolled TD patients assigned to the non-TD group, and 148 healthy persons assigned to the normal group. The gene distribution rate of Val9Ala gene was analyzed using polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) analysis, and the therapeutic effect of CTD on 36 patients with TD was observed after 12 weeks treatment.

RESULTSThere was no significant difference in genotype and allelic gene frequency of SOD Val9Ala among the TD, non-TD and normal groups (P > 0.05). Comparison of the AIMS score in TD patients with various Val9Ala genotypes showed that the difference of AIMS scores in patients with TT and CT genotype was not significant (P > 0.05), but CTD did show a better efficacy in TD patients with CT heterozygote than in those with TT homozygote (P < 0.05).

CONCLUSIONThe CTD could effectively relieve the symptoms of TD, its efficacy might be related with the genotype of SOD, and 9Ala is considered to be a protective factor for the susceptibility to TD.

Adolescent ; Adult ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Dyskinesia, Drug-Induced ; drug therapy ; etiology ; genetics ; Female ; Humans ; Male ; Middle Aged ; Phytotherapy ; Polymorphism, Genetic ; genetics ; Superoxide Dismutase ; genetics ; Valine ; genetics ; Young Adult

OBJECTIVETo evaluated the cytotoxicity of a new nano-hydroxyapatite (n-HA) root canal sealer.

METHODSIn this study, the cytotoxicity was evaluated by the method of MTT assay in vitro, and culture medium F12 as control, three concentrations of the soaking material cultured with mouse osteoblast separately, to test the cell relative growth rate (RGR) of every group.

RESULTSThe toxicity graduation of the n-HA root canal sealer tends to 0 with the culture time increasing. The cell survival rate of n-HA root canal sealer showed high relatively. The OD value of cell was similar for the negative control and the extracts (P > 0.05).

CONCLUSIONThe result indicated that n-HA root canal sealer was compatible with the testcells.

Animals ; Cell Survival ; Dental Pulp Cavity ; Durapatite ; Mice ; Root Canal Filling Materials ; Zinc Oxide-Eugenol Cement

Objective:To investigate the association between dopamine receptor D2(DRD2) polymorphisms and smoking in male patients with schizophrenia.Methods:Totally 773 patients with schizophrenia (567 smokers and 206 non-smokers) and 302 normal controls (168 smokers and 134 non-smokers) were recruited.The two single nucleotide polymorphisms (SNPs) (rs1800497 and rs1079597) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RELP).SHEsis genetic analysis platform was used to calculate linkage disequilibrium index and infer allele distribution and haplotype frequency.Results:There was no significant difference in two SNPs genotype and allele distributions between the patients and normal controls or between smokers and non-smokers in either patients or normal controls alone (Ps ＞ 0.05);the frequency estimations of haplotype C-A and T-G in patients with schizophrenia were higher than in normal controls (8.0％ vs.5.2％,10.2％ vs.4.1％,Ps ＜0.05),T-A (34.6％ vs.40.2％,P ＜0.05),whereas the frequency estimation of haplotype T-A in patients with schizophrenia was lower than in normal controls,and all the differences were statistically significant (34.6％ vs.40.2％,P ＜ 0.05).It was also observed that the frequency estimation of haplotype T-A in normal smokers was significantly lower than in normal non-smokers (2.5％ vs.6.1％,P ＜0.05).Conclusion:There may be a correlation between DRD2 polymorphisms and the susceptibility to schizophrenia,but not between DRD2 polymorphisms and smoking neither in patients with schizophrenia nor in normal controls.

Animals ; Autoantibodies ; blood ; Cytokines ; biosynthesis ; Dose-Response Relationship, Drug ; Female ; Immunohistochemistry ; Rats ; Rats, Inbred Lew ; Selenium ; administration & dosage ; therapeutic use ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology ; Thyroid Gland ; drug effects ; immunology ; pathology ; Thyroiditis, Autoimmune ; drug therapy ; immunology ; Trace Elements ; administration & dosage ; therapeutic use

A novel dual luciferase expression vector was designed and its expression characteristics were studied in vitro and in vivo. Firstly, the Gluc and Fluc genes were connected via the TaV 2A sequence by overlaping PCR, and inserted into the expression vector pAAV2neoCAG, obtaining the recombinant plasmid pAAV2neoCAG-Gluc-2A-Fluc. Then pAAV2neoCAG-Gluc-2A-Fluc was transfected into BHK21 cells and the activity of Gluc and Fluc in the supernatant and cell lysates were assayed respectively. Results showed that both Gluc and Fluc were expressed successfully. The Gluc was mainly detected in the culture media while the Fluc was mostly within cells. The activity of Gluc in the supernatant increased gradually with time while the Fluc activity in cells almost kept stable. To investigate the expression of pAAV2neoCAG-Gluc-2A-Fluc in vivo, the plasmid was hydrodynamically injected into BALB/c mice through tail vein. The Gluc activity was assayed in a small volume of blood taken by tail vein at different time points. Results showed that Gluc was expressed stably at least 7 days. Live bioluminescence imaging technology was used to compare the expression characteristics of Gluc and Fluc. Whole body imaging was seen when coelenterazine, a specific substrate for Gluc, was injected, and the imaging signals decreased rapidly within 10 minutes. Liver imaging was showed when Flue specific substrate named D-Luciferin was injected, and the imaging remained stable at least for half an hour. The dual luciferase expression vector pAAV2neoCAG-Gluc-2A-Fluc combines the advantages of the secreted report gene Gluc and the non-secreted report gene Fluc, and will provide a new tool for cell labeling and tracing.
Animals ; Base Sequence ; Cricetinae ; Crustacea ; enzymology ; Fireflies ; enzymology ; Gene Expression ; Genes, Reporter ; Genetic Vectors ; genetics ; metabolism ; Luciferases ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data

OBJECTIVETo study the occurrence of Cercospora leaf spot of Lonicera macrathoides and fungicides control method, so as to provide scientific basis for its integrated pests management (IPM).

METHODThe field investigation and the field controlling trial were carried out for the research.

RESULTCercospora leaf spot was caused by C. rhamni. There was obvious relationship between the damage rate and the altitude, soil organic matter, available K, available P as well as variety. The controlling effect of 10% difenoconazole WG was the highest. 50% Thiram WP, 70% Mancozeb WP also had well controlling effect.

CONCLUSIONCercospora leaf spot of L. macrathoides could be controlled by suitability agricultural and chemical controls.

Ascomycota ; drug effects ; isolation & purification ; pathogenicity ; physiology ; Fungicides, Industrial ; pharmacology ; Lonicera ; microbiology ; Plant Diseases ; microbiology ; prevention & control ; Plant Leaves ; microbiology ; Virulence

Aim To investigate the neuroprotective effects of puerarin on H2O2-induced SH-SY5Y cell ap-optosis and the molecular mechanisms underlying the neuroprotective effects. Methods Neuron injury mod-el was established in vitro through H2O2-induced SH-SY5Y injury. MTT assay was performed to detect the effect of puerarin on H2O2-induced SH-SY5Y survival rates. Hoechst 33342 staining was used to observe the cell apoptosis. JC-1 staining was employed to detect the level of mitochondria membrane poential. Caspase-3 was determined by caspase-3 catalyze the substrate specificity Ac-DEVD-pNA. Caspase-9 was determined by caspase-9 catalyze the substrate specificity Ac-LE-HD-pNA. The effects of puerarin on the protein level of Bcl-2,Bax,p-Akt and Akt were determined by West-ern blot. Results The cell survival rate significantly increased after puerarin pretreatment compared with H2O2model group. Furthermore, puerarin pretreat-ment not only inhibited the decreasing of mitochondrial membrane potential,increasing of caspase-3, caspase-9 enzymatic activity and the expression of Bax,but also promoted the expression of p-Akt and Bcl-2, which was prevented by LY294002, an inhibitor of PI3K/Akt. Conclusion Puerarin can play a neuroprotective role for SH-SY5Y cell apoptosis induced by H2O2, maybe via activating PI3K/Akt signaling pathway.