OBJECTIVE:To establish a method for the contents determination of gallic acid,naringin,hesperidin,neohesperi-din and baicalin in Qinghou liyan granule. METHODS:UPLC was performed on the column of ACQUITY UPLC BEH C18 with mobile phase of acetonitrile- 0.1% phosphoric acid(gradient elution)at a flow rate of 0.40 ml/min,detection wavelength was 280 nm,column temperature was 30 ℃,and injection volume was 1 μl. RESULTS:The linear range was 0.048-0.480 μg for gallic ac-id(r=0.999 3),0.012-0.120 μg for naringin(r=0.999 9),0.016-0.160 μg for hesperidin(r=0.999 8),0.022-0.220 μg for neo-hesperidin(r=0.999 9)and 0.072-0.720 μg for baicalin(r=0.999 2);limit of quantitation was 2.4,1.6,1.8,1.8,1.6 ng,limit of detection was 7.5,5.0,5.6,5.6,5.0 ng;RSDs of precision,stability and reproducibility tests were no lower than 3.0%;recover-ies were 96.64%-102.02%(RSD=2.00%,n=6),95.86%-102.56%(RSD=2.86%,n=6),97.24%-102.54%(RSD=2.10%,n=6),97.44%-102.60%(RSD=2.40%,n=6)and 96.91%-103.13%(RSD=2.62%,n=6). CONCLUSIONS:The method is rapid and accurate,and can be used for the simultaneous determination of gallic acid,naringin,hesperidin,neohesperidin and baicalin in Qinghou liyan granule.

Objective To detect levels of C?X?C chemokine receptor type 5 (CXCR5) and inducible costimulator(ICOS)in blister fluid of patients with bullous pemphigoid(BP), and to explore their significance in the pathogenesis of BP. Methods Blister fluid samples were collected from 15 patients with BP(experimental group)and 15 patients with second?degree burns(control group). Enzyme?linked immunosorbent assay(ELISA)was performed to detect the levels of CXCR5 and ICOS in the 2 groups. Results The level of CXCR5 was significantly higher in the experimental group than in the control group(219 ± 145.31 vs. 147 ± 23.83 ng/L, t=4.577, P<0.05), while no significant difference in the ICOS level was observed between the 2 groups (30.18 ± 14.86 vs. 21.43 ± 5.32 ng/L, t = 1.628, P > 0.05). Conclusion The expression of CXCR5 may be associated with the occurrence of BP, but further researches are needed to determine the relationship between ICOS and the occurrence of BP.

BACKGROUNDTo determine the antigenicity of HGV NS5 recombinant proteins expressed in E.coli.

METHODSHGV NS5a,NS5b and core/NS5b fusion genes were cloned into pThioC vector. Three expression plasmids were transformed into JM109(DE3) competent cells then expressed with induction by IPTG. Western blot and ELISA were used to determine the antigenicity after the three recombinant proteins were purified.

RESULTSAfter identification by restriction enzyme and sequencing, it was confirmed that the expressed was target proteins espected. Purified expression proteins were found strongly immunoreactive among anti HGV positive sera by Western blot and ELISA. Compared with mixed recombinant antigen (including core, NS5a synthetic peptide and NS3 recombinant proteins), in the 22 positive sera detected with mixed antigen, 68%(15/22), 90%(20/22) and 73%(16/22) were positive by P5a,P5b and Pc?5b antigens; In the 70 negative samples with mixed antigen, 7%(5/70), 1%(1/70) and 6%(4/70) were positive by P5a, P5b and Pc?5b antigens. The positive alone was found among RTPCR positive specimen using these recombinant antigens.

CONCLUSIONSNS5 gene expressed in E.coli?which couldn't be covered with other regions of antigens was one of the essential epitopes to HGV immunologic diagnosis.

Antibodies, Viral ; blood ; Antigens, Viral ; blood ; Epitopes ; immunology ; GB virus C ; genetics ; immunology ; Humans ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; immunology ; Viral Nonstructural Proteins ; genetics ; immunology

Objective: To study the effect of 60Co-γ ray irradiation on the contents of danshensu,protocatechuic aldehyde,puerarin and salviamolic acid B in Tongmai granules by HPLC. Methods: An Agilent ZORBAX Eclipse XDB C18(2) column (250 mm × 4. 6 mm,5 μm) was adopted and the wavelength of UV detection was 280 nm at the flow rate of 1. 0 ml·min-1. The mobile phase consisted of 0. 1% phosphoric acid (B) and acetonitrile (A) with gradient elution, and the column temperature was 35℃. Tongmai granules were irradiated by 60Co-γ ray respectively at 0, 2, 5,10 and kGy,the contents of the active ingredients were compared before and after the irradiation. Results: The linear range of danshensu,protocatechuic aldehyde,puerarin and salviamolic acid B was 0. 098-4. 925 μg, 0. 028-1. 411 μg, 0. 378-18. 882 μg and 0. 218-10. 888 μg, respectively. The average recovery was 99. 8% , 97. 7% , 99. 9% and 99. 9% , respectively. When the radiation dose was not more than 2 kGy, the contents of the five components did not change significantly (P>0. 05). After 5 kGy radiation, the contents of protocatechuic aldehyde and salviamolic acid B were signifi-cantly different (P <0. 05). Conclusion: The dose of 60Co ray should be controlled not more than 2 kGy, and the sterilization method is safe and effective for Tongmai granules.

Organoid is a newly developed cellular there-dimensional culture system in recent years. Organoids have a three-dimensional structure, which is similar to that of the real organs. Together with the characteristics of self-renewal and reproduction of tissue origin, organoids can better simulate the function of real organs. Organoids provide a new platform for the study of organogenesis, regeneration, disease pathogenesis, and drug screening. The digestive system is an essential part of the human body and performs important functions. To date, organoid models of various digestive organs have been successfully established. This review summarizes the latest research progress of organoids of taste buds, esophagi, stomachs, livers and intestines, and prospects future application of organoids.
Humans ; Organoids ; Intestines ; Liver

Extracellular matrix is the main element to provide mechanical clues for cells. The response of stem cells to mechanical signals is mainly achieved through the cytoskeleton. After mechanical signal is transmitted, cytoskeleton can form contractile microfilaments that actively generate tension through reorganization induced by microenvironment changes. The mechanical signals can regulate gene expression through either coupling with the nuclear skeleton directly or being transformed by the second message. Recent studies have proven that cytoskeleton tension has a series of impact on lineage specification, proliferation, differentiation and apoptosis of bone mesenchymal stem cells (BMSCs). BMSCs are of great significance in bone reconstruction and clinical treatment. The possible mechanisms about mechanotransduction and its effects of cytoskeleton tension on osteogenesis of BMSCs after micro-environmental changes were summarized.