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.

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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.

Objective To express and purify recombinant and biologically active Clostridium difficile toxin B (rTcdB). Methods The genes of TcdB were amplified by polymerase chain reaction (PCR) using chromosomal DNA from a toxigenic strain, and cloned into a shuttle vector pHis1522.The sequences of TcdB genes in the vector were verified by DNA sequencing. The construction was transformed into Bacillus megaterium protoplasts and the protein expression was driven by a xylose promoter. The purified protein was tested for biological activity. Results rTcdB was successfully purified from bacterial crude extracts. Approximately 5-10 mg of highly purified recombinant toxin was obtained from one liter of bacterial culture. The expressed rTcdB had molecular mass similar to the native toxin, and its biological activity was proved to be similar to its native counterpart after an extensive examination. Conclusion rTcdB with biological activities is successfully expressed in Bacillus megaterium.

Background Mutant C57BL/6 mouse with corneal opacity (B6-Co) appears eye open at birth (EOB) phenotype,which is a good animal model in the study of developmental mechanism of eyelid.Investigating the relationship between serum response factor (SRF) and EOB phenotype can provide theoretical support for the research on the mechanism of innate defects in eyelid development in humans.Objective This study was to assess the dynamic expressions of SRF in eyelid of embryonic B6-Co mouse.Methods Total RNA was extracted from B6 and B6-Co mice eyelid tissue at embryonic day 16.5 (E16.5 d),E17.5 d and E18.5 d.The relative expression levels of SRF mRNA and protein in the eyelid tissue of B6 and B6-Co embryonic mice were assayed by real-time quantitative PCR and Western blot,respectively.In situ expressions of SRF protein in eyelid of B6-Co mice and B6 mice were detected using immunofluorescence technique.The use and care of the animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals of Nantong University.Results The relative expression levels ofSRF mRNA in the eyelids were 0.41±0.06 and 0.24±0.17 in E16.5 d and E17.5 d of B6-Co mice,showing a significant decline in comparison with 1.03 ±0.17 and 1.01 ±0.09 in the B6 mice (P =0.025,0.017).The expression levels of SRF protein in the eyelids of E16.5 d and E17.5 d B6-Co mice were 0.08±0.01 and 0.08± 0.01,which were significantly lower than 0.12 ±0.03 and 0.13 ± 0.02 of B6 mice (P =0.036,0.024).However,there were no significant differences in the expression levels of SRF mRNA and protein in E18.5 d between the B6-Co mice and B6 mice (P =0.387,0.774).Immunofluorescence assay displayed that SRF was expressed in the keratinocytes of eyelids in both mice,but the fluorescence intensity was weaker in the B6-Co mice.Conclusions SRF probably interrupts the developing process of eyelid in early embryo of B6-Co mice.

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.

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 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 analyze the differences of adverse drug reactions between children and adults caused by Tanreqing injection,and provide reasonable application of Tanreqing injection.Methods Retrospective research method were adopted to screen out the adverse drug reaction reports caused by Tanreqing injection from drug ADR reports of Drug ADR Monitoring Center affiliated to the Second People's Hospital of Gansu Province from January 2010 to October 2012.Figures of children and adults who used Tanreqing injection were analyzed separately.Results A total of 603 children used Tanreqing injection,among which 5 children appeared ADR (0.83％).A total of 4395 adult used of Tanreqing injection,among which 4 appeared in ADR (0.09％).There was statistical difference between the two groups (x2 =16.07,P＜ 0.05) The incidence rate of ADR in males was more than females either in the adults or in children.Hypersensitivity was the most common ADR.Conclusion ADR of Tanreqing injection in Children was significantly higher than adults.Tanreqing injection should be used carefully and rationally.

Objective To explore current status of health-related productivity loss and its risk factors among nurses.Methods Stanford presenteeism scale (SPS-6) and self-designed questionnaire were used to investigate current status of health-related productivity loss and its risk factors among 1122 nurses working in a tertiary hospital in Changchun city.Results Compared with hired nurses,age and work seniority of permanent nurses were significantly higher(Z =-19.49,-19.28 ;P ＜0.05).The average score of SPS-6 of all the participants was 20.05 ± 4.37.The score of SPS-6 of married nurses was significantly lower than other nurses (Z =-3.52,P ＜ 0.05) ; and the score of SPS-6 of nurses less than 30 years old was significantly higher than those above 30 years old (Z =-2.49,P ＜ 0.05).There were no significant differences between the SPS-6 score of education degree and department.(Z =-1.37,x2 =0.58 ; P ＞0.05).The result of GLM showed that employment status and work seniority were independent risk factors of health-related productivity loss among nurses.The scores of SPS-6 of permanent nurses was significantly lower than hired nursed (x2 =4.48,P ＜ 0.05),and those who had worked for less than 3 years showed significantly higher score of SPS-6 than those who had worked longer (x2 =12.89,P ＜ 0.05).Conclusions Health-related productivity loss do exist among nurses.Improving health management may help to reduce this loss of productivity.