Hirudo nipponica Whitman,a Chinese medicine which has a strong effect on platelet aggregation and antithrombin activity,is involved in the studies.The current situation is adverse to the cultivation,development and utilization of H.nipponica.Prior to summarizing the current research status and the unresolved problems and the prospective of H.nipponica,domestic and foreign literatures over the H.nipponica was discussed and judged,aiming at providing a reference for the comprehensive development and utilization of H.nipponica.

The pharmaceutical microbiological examination in medical institutions is an important method to ensure the safety of preparations.It is of great significance to popularize the techniques of drug microbiological examination in primary medical institutions.This article started with the purpose, plan and training contents of the microbiological examination training program, which combined with theory teaching and practical operation, two-phase practical teaching, group teaching and collective teaching and guided students to design and plan experiments to carry out the teaching work,and then explored the ways and improving measures of the pharmaceutical microbiological examination training for hospital pharmacists, which could provide reference for the training and teaching of hospital pharmaceutical microbiological examination.

In order to improve the production of agro-antibiotic 2-16,the producing strain(Streptomyces ahygroscopicus var.huangshanensis) was treated by protoplast regeneration,ultraviolet radiation,NTG mutagenesis and low energy C~(+) ion implantation.At last,a high-yield strain No.515 was obtained.The production of ~()No.515 was increased by 223.10%.By using Plackett-Burman design and Response Surface Analysis provided by SAS software,the cultivation condition of No.515 was optimized.The amount of agro-antibiotic 2-16 was increased by 38.53% when the strain No.515 was cultivated in the optimum medium instead of the initial one.

To address the role of Pr1 gene in the process of Hirsutella sinensis infecting Hepialus gonggaensis, differential expression of subtilisin-like protease gene was detected. In the present study, Pr1 gene analogues from H. sinensis were obtained by PCR strategy using specific primers designed from conserved regions of Pr1 gene reported in the GenBank. Then we detected the changes in the expression of Pr1 gene before and after infecting H. gonggaensis using real-time quantitative PCR. We obtained the partial sequence of Pr1 gene with the length of 535 bp (GenBank accession: KC009680). Real-time PCR results showed that the expression level of Pr1 gene was significantly different among 8 samples (P < 0.01). Pr1 gene showed the obvious higher expression level (2-3 folds) after infecting the H. gonggaensis, suggesting that the Pr1 gene may play an important role in the process of H. sinensis infecting H. gonggaensis. The present study paves a way for further identification on infectivity assessment of H. sinensis.
Amino Acid Sequence ; Animals ; Hypocreales ; genetics ; metabolism ; pathogenicity ; Larva ; Lepidoptera ; microbiology ; Real-Time Polymerase Chain Reaction ; Subtilisin ; genetics ; metabolism

Objective To explore the anti-inflammatory and antipruritic effects of Chushizhiyang ointment in a mouse model. Methods A total of 40 male 8-week-old BALB/c mice were included in this study, and randomly and equally divided into 4 groups. A mouse model of atopic dermatitis (AD)was established in three groups of mice by repeated application of 2,4-dinitroflurobenzene (DNFB)to shaved abdominal skin for sensitization and to shaved dorsal skin for stimulation. After establishment of the AD model, the three groups were topically treated with sodium chloride physiological solution (model group), hydrocortisone cream (hydrocortisone group)and Chushizhiyang ointment (Chushizhiyang group)respectively for 14 consecutive days. The remaining group receiving no sensitization or treatment served as the normal control group. All the mice were sacrificed 12 hours after the final treatment, and the dorsal skin of mice was resected followed by the determination of skin thickness and weight as well as hematoxylin-eosin(HE)staining and toluidine blue staining for the counting of leukocytes and mast cells respectively. Moreover, enzyme-linked immunosorbent assay(ELISA)was performed to measure the levels of interferon-gamma(IFN-γ), tumor necrosis factor alpha(TNF-α), interleukin 4(IL-4)and IL-5 in dorsal skin lesions. In addition, a local skin itching model was induced by histamine phosphate in Hartley guinea pigs, which was used to explore the effect of Chushizhiyang ointment on itch thresholds. Results Compared with the model group, both the Chushizhiyang group and hydrocortisone group showed reduced thickness and weight of dorsal skin in mice (all P < 0.01), numbers of infiltrating lymphocytes and mast cells (all P < 0.01)and levels of IFN-γ, TNF-α, IL-4 and IL-5 in skin lesions (P < 0.05 or 0.01)on day 15 after the start of treatment. The thickness and weight of dorsal skin in mice were significantly decreased in the hydrocortisone group (P <0.01), but experienced no significant changes in the Chushizhiyang group compared with the normal control group. Additionally, Chushizhiyang ointment could significantly increase itch thresholds in guinea pigs induced by histamine phosphate(P < 0.01). Conclusions Chushizhiyang ointment can significantly inhibit DNFB-induced AD in mice, likely by restoring the balance between Th1 and Th2 type cytokines. Moreover, Chushizhiyang ointment could markedly relieve itching induced by histamine phosphate in guinea pigs.

PurposeMultiple sclerosis (MS) is characterized by time and spatial multiple, and it is the main reason for disabled young people. This paper aims to investigate the application of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) with dual-compartment Tofts model in relapsing-remitting multiple sclerosis (RRMS) and its correlation with clinical scoring.Materials and MethodsThe clinical data of 25 patients with RRMS were retrospectively studied. The patients underwent the conventional MRI and the DCE-MRI examination. The result was processed by dual-compartment Tofts model and quantitative measurement was carried out in terms of volume transfer constant (Ktrans), rate constant between EES and blood plasma (Kep) and the volume of EES per unit volume of tissue (Ve), cerebral blood flow (CBF) and cerebral blood volume (CBV) of the lesions and normal-appearing white matter (NAWM) regions. The correlation between imaging biomarkers, expanded disability states scale (EDSS) and disease duration were also analyzed.Results ① The differences of MR imaging biomarkers Ktrans and Kep were significant between the regions of nonenhancing (NE) lesions, the NAWM regions near NE lesions and the NAWM regions far from NE lesions (χ2=6.777 and 22.343,P<0.05); however, Ve in the NE lesions had no significant differences compared with that in the NAWM regions near and far from NE lesions (P>0.05).②The CBF and CBV among these three groups had no signiifcant differences (P>0.05).③The CBF of NE lesions was signiifcantly correlated with disease duration (r=0.518,P<0.05);however, the other markers like Ktrans, Kep, Ve, CBF and CBV were neither signiifcantly correlated with EDSS nor with disease duration (r=-0.371-0.052,P>0.05).Conclusion DCE-MRI with Tofts model can quantitatively measure microvascular permeability and perfusion characteristics of lesions and NAWM regions, which thus reflects hemodynamic changes in patients with multiple sclerosis.

Adolescent ; Adult ; Asian Continental Ancestry Group ; Carcinoma, Hepatocellular ; genetics ; virology ; Ethnic Groups ; Female ; Genotype ; Hepatitis B ; genetics ; virology ; Hepatitis B virus ; genetics ; Humans ; Liver Cirrhosis ; genetics ; virology ; Liver Neoplasms ; genetics ; virology ; Male ; Middle Aged ; Mutation ; Young Adult

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua ; genetics ; metabolism ; Artemisinins ; metabolism ; Biosynthetic Pathways ; Drugs, Chinese Herbal ; Mixed Function Oxygenases ; genetics ; Oxidoreductases ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism

The present study investigated the relationship between DNA-dependent protein kinase (DNA-PK) and radiosensitivity of nasopharyngeal carcinoma (NPC) cell lines. The dose-survival relationship for NPC cell lines, CNE1 and CNE2, was analyzed using clonogenic formation assay, the activity of DNA-PK of the two cell lines was measured using the Signa TECT DNA-PK assay kit, and the localization and expression of Kus (a heterodimer) and DNA-PKcs protein in CNE1 and CNE2 before irradiation and 15 min, 1 h, 6 h, 12 h, 24 h after 4 Gy irradiation were analyzed by immunofluorescence, laser scanning confocal microscope (LSCM) and Western blot. The results showed that the surviving fraction of CNE1 was higher than that of CNE2 at each dose. The DNA-PK activity of CNE1 was also significantly higher than that of CNE2 before and after irradiation (P<0.05), while the expression of total Ku70/Ku80 in CNE1 and CNE2 had no significant difference. Increasing translocation of Ku70 and Ku80 from the cytoplasm to the nuclei in the two cell lines was observed with increase of irradiation time as detected by Western blot, and the immunofluorescence of the DNA-PK complex subunits showed greater nuclear translocation in CNE1 than CNE2 after irradiation. The results suggest that the relatively higher radio-resistance of CNE1 correlates with the higher activity of DNA-PK as compared to that of more radiosensitive CNE2 (or lower radio-resistance) before and after irradiation. Thus, DNA-PK activity may be a useful predictor of radiosensitivity of NPC.
Carcinoma ; Cell Line, Tumor ; enzymology ; radiation effects ; DNA-Activated Protein Kinase ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; enzymology ; Radiation Tolerance