Objective:To simultaneously determine the contents of ginsenoside Rb1 , ginsenoside Rb2 , ginsenoside Rb3 , ginsen-oside Re, ginsenoside Rg1 , ginsenoside Rf and ginsenoside Ro in Qipi pills by HPLC-QTOF-MS. Methods: The determination was performed on an Agilent Poroshell 120 EC-C18 column (2. 1 mm × 50 mm,2. 7 mm) with mobile phase consisting of acetonitrile( A)-water(B, containing 0. 1% formic acid) with gradient elution. The flow rate was 0. 21 ml·min-1. The column temperature was 30℃. The MS instrument was equipped with an ESI+ ion source. The exacted ion chromatograms were used to determine the quantities of different compounds in the samples while the mass spectra of product ions were used for confirming the compounds. Results:All the 7 kinds of ginsenoside showed good linearity (r>0. 9993). The RSDs of precision, repeatability and stability tests were all less than 5%. The average recoveries were within the range of 97. 11%-101. 98%. Conclusion:The method is simple, rapid and reliable with high specificity, which can be used for the quality control of Qipi pills.

With the development of imaging techniques, aphasia caused by subcortical impairment is increasingly found in recent years. This paper reviewed the literature about the subcortical aphasia and expolred the anatomical and clinical features and pathogenesis of subcortical aphasia.

Objective To construct Cchl1a3 gene R528H knock-in mouse model related to hypokalemic periodic paralysis.Methods ES cells were transfected with Cchl1a3-Konckin targeting vector linearized by Not I digestion , selected in the medium containing both G 418 and ganciclovoir .Resistant clones were screened by PCR and further confirmed by DNA sequencing for correct homologous recombinants .Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts .Heterozygous mice were obtained by mating .Through heterozygous mice with FLP mice mating , removal of neo gene heterozygous mice were established and identified with the PCR and DNA sequencing . After mating, homozygous offspring were constructed and observed .Results ES cells were successfully transfected withtargeting vector .It was confirmed that 9 resistant clones happened right homologous recombination by PCR and DNA sequencing .7 chimera mice were obtained by microinjection .After breeding the chimeric mice , heterozygous mice were mated FLP mice to obtain 9 heterozygous mice removal of neo gene, the finally obtained 15 homozygous mice with Flp-deleted neo gene.In the developmental stage of sexual maturity , the spirit of the mice, restaurants and activities in good condition, but the gradual emergence of hair removal at 4 months of age, skin ulceration and even death .Conclusions We successfully constructed Cchl1a3 gene R528H mutation homozygous mice.And it laid a foundation for the study of human CACNA1S gene function and to clarify the molecular mechanism of hypokalemic periodic paralysis .

In order to culture the qualified clinicians, we should think about how to improve the quality of clinical practice of obstetrics and gynecology. It is of great importance to emphasize the teachers and students to value the teaching work together. Importance should be attached to the advantage of subject of academy, to make the clinical practice closer to practical, which has a perfect effect and will benefit our work.

Purpose　The aim is to prepare the extract of immunocompetent cell proliferation from human placenta and to try to find out a suitable method of preserving the extract.Methods　The extract was mainly prepared by heat-treating placenta homogenated fluid. Then the activity to stimulate murine splenic lymphocyte proliferation in vitro was done with MTT , exposing　the　extract to　radioisotope 60　Co.Results　The content of protein was 2～3mg per gram placenta measured by Lowry′s Method.The rate to promote cell proliferation was more than 80 percent.The activity lasted a few months after being exposed to radioisotope.Conclusion　The extract prepared by heat-treating not only had high activity, but also had the unique method and good repeatability.This prepared extract as a kind of stable,reliable and remarkably promoting lymphocyte proliferation reaction had the value of development production on broad scale as well as in clinical practice.

Objective To prepare the human bone sialoprotein (BSP) monoclonal antibodies (mAb)with high titer and specificity and identify its characterization,which is based on further studying BSP as clinical biomarker for breast cancer metastasizing to bone. Methods BALB/c mice were immunized with purified recombinant BSP protein.Cell fusion was performed between mouse splenic cells and myeloma cells (Sp2/0), and then the hybridoma cell lines secreting mAb against BSP antigen were screened and cloned. The ascites were prepared and purified with Protein G affinity chromatography.The titer and subtypes of mAb against BSP were identified and measured by ELISA and Western blotting analysis. ResultsNine hybridoma cell lines that stably secreted mAb against BSP were successfully obtained.Two of them,D001 and D002,were further identified, which belonged to the subtypes of IgG1 and κ light chain. The two antibodies titers in culture supernatant were 1∶5120 and 1∶10 240, respectively, and those in the ascites fluid were 1∶25 600 and 1∶51 200,respectively.Results of Western blotting analysis and immunohistochemistry showed that the two antibodies could specifically bind with BSP derived from human breast cancer cells.ConclusionNine mAb against BSP have been successfully prepared which can be used for further studying the biological properties of BSP and reveal its relationship with data from clinic patients.

This study aimed at exploring the features of the body constitution and syndrome types of TCM in patients with pSS.General information of 60 patients with pSS was collected.Their TCM syndrome types and TCM body constitution types were examed,while the distribution of pSS syndrome types in terms of their body constitution was analyzed.Among 60 pSS patients,58 of them (96.67％) were female with the average age of 52 years and an average course of 63 months.Their usual residence was Xinjiang.Their most common first symptoms included thirst,dry eyes and joint pain,and the damage of skin and mucous membranes and blood system was rare.It was found that the syndrome of both qi and yin deficiency was the most common (48.33％),followed by the impregnation syndrome of heat and dampness (26.67％) and the syndrome of the weakness of spleen qi (25％) in pSS patients.Yin deficiency (58.33％) and yang deficiency (58.33％) in the body constitution of TCM accounted for decent proportions,followed by the constitutions of qi stagnation (48.33％),qi deficiency (45.00％),blood stasis (38.33％),phlegm-dampness (36.67％),damp-heat (18.33％),allergic (13.33％) and yin-yang harmony (1.67％).Those without the health education of pSS accounted for 51.11％ in cases of palindromia.In conclusion,yin deficiency and yang deficiency constitutions were most common,at most,the body constitution of both qi and yin deficiency,in pSS patients in accordance with TCM body constitution.Most pSS patients of palindromia didn't received professional health education of pSS.All the causes above suggested that we should make individualized treatment programs according to clinical syndrome types and the characteristics of TCM body constitution in pSS patients for strengthening the health management of pSS and improving their quality of life.

Objective: To explore the role of calcium/calmodulin-dependent protein kinase-II δ (CaMK-II δ) in doxorubicin (DOX) induced cardio-toxicity in experimental rats.
Methods:①The rat’s cardiomyocytes were treated by DOX and the cell proliferation, protein expression and activity of CaMK-II δ were examined.②CaMK-II δ gene was knocked out by CRISPR method, the changes of DOX induced cell apoptosis and NF-κb activity and miR-146a expression were detected.
Results: DOX could inhibit cardiomyocyte proliferation, the protein expression level of CaMK-II δ was similar and the activity was increased. CRISPR method may effectively knock out CaMK-II δ gene. Compared with normal cells, the cells from CaMK-II δ knocked out rats had decreased sensitivity to DOX induction, suppressed NF-κb activation and miR-146a up-regulation.
Conclusion: CaMK-II δ participated in DOX induced cardio-toxicity in experimental rats and NF-κb and miR-146a were involved in this process.

Objective To observe the ability of inactivating human immunodeficiency virus type 1 (HIV-1) in whole blood by visible light combination with methylene blue (MB) at different concentrations. Methods HIV-1 was used as the test virus. The contaminated blood was treated by MB, visible light (640 nm), combined with MB and visible light (640 nm). The test of MT4 cell infection was used to evaluate the virus inactivation efficacy. Results After being treated by MB at the concentrations of 5, 10, and 15 ?mol/L, and then irradiated respectively by visible light (40 000 Lux, 640 nm) for 30, 20, and 10 min, all the added indicated virus at the titer of 10 5.78 TCID 50 of HIV-1 could be inactivated absolutely. Conclusion Methylene blue photochemical method can inactivate HIV-1 in blood effectively.

Objective:To develop a method of quantitative analysis of multi-components by single marker( QAMS) for nine kinds of alkaloids in Xiaohuoluo pills. Methods: An HPLC-QTOF-MS method with an Agilent ZORBAX Extend-C18 RRHT(2. 1 mm × 50 mm,1. 8 μm) column was applied. The flow rate was 0. 21 ml·min-1 . The column temperature was 30 ℃. The mobile phase was methanol (A)-water (B;containing 0. 1％ formic acid and 2. 5 mmol·L-1 ammonium acetate) with gradient elution. The aconitine was used as the internal standard, and the relative correction factor ( RCFs) of hypaconitine, mesaconitine, benzoylaconine, benzoyl-hypaconine, benzoylmesaconine, aconine, hypaconine and mesaconine was respectively established, and the reproducibility inspection on the RCF was performed. The contents of the other 8 kinds of aconitum alkaloids were calculated according to the RCF. At the same time, an external standard method ( ESM) was performed for the content determination of the nine alkaloids. The results of the two methods were compared. The feasibility and accuracy of the QAMS method were verified. Results:Within a certain range,the RCF of hypacontine,mesacontine, benzoylaconine, benzoylhypaconine, benzoyl mesaconine, aconine, hypaconine and mesaconine to aconitine was 1. 736,1. 979,1. 0471,0. 9242,1. 2901,1. 3078,1. 2859,and 1. 0948,respectively. The QAMS method was established for determi-ning alkaloids. There were no significant differences between the results of the QAMS method and those of the external standard method ( ESM) . Conclusion:With the validation of methodology, the method established in our study can be used for the content determina-tion of aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, aconine, hypaconine and mesaconine in xiaohuoluo pills.