Objective To express nucleoprotein(NP) of influenza A virus in Bac-to-Bac TOPO system and evaluate its immunogenicity,so as to lay a foundation for the research of influenza virus NP protein vaccine.Methods The NP gene of influenza A H1N1(A/Guangdong-Maonan/SWL1536/2019_CNIC-1909) was amplified by PCR and connected to the vector pFastBac~(TM)/CT-TOPO~(TM),which was transformed into competent E.coli One Shot~(TM) Mach1~(TM) T1~R to prepare donor plasmid,and identified by PCR and sequencing.The donor plasmid was transformed into competent E.coli MAX Efficiency~(TM) DH10Bac~(TM),the positive monoclone was screened by blue-white spot screening,and the recombinant bacmid was extracted and identified by PCR.The recombinant bacmid was transfected into Sf9 insect cells to prepare the recombinant baculovirus,and the virus titer was detected by flow cytometry.After multiple rounds of amplification,the recombinant baculovirus infected Sf9 insect cells for the expression of the target protein NP,and the recombinant protein was purified by nickel ion affinity chromatography.Female BALB/c mice were subcutaneously inoculated with recombinant NP protein and PBS respectively,with six mice in each group.The serum specific antibody levels of mice were detected by ELISA,the IFN-γ secreted by spleen cells were detected by flow cytometry,and the CD4~+ and CD8~+ lymphocytes produced by spleen cells were detected by ELISpot.Two weeks after the last immunization,the mice were challenged with A/Guangdong Maonan/SWL 1536/2019_CNIC-1909 influenza virus through nasal cavity,and the body mass changes and survival rate of mice within 14 days after challenge were monitored.Results The results of PCR and sequencing showed that the donor plasmid was constructed correctly.PCR identification results confirmed that the recombinant bacmid was successfully constructed.The titer of recombinant baculovirus was 2.875 × 10~8ivp/mL.The recombinant NP protein showed specific binding to mouse anti-influenza A virus NP monoclonal antibody,with the purity of 91.3% after purification.Two weeks after the last immunization,compared with the PBS group,the serum specific antibody titer(t=0.288,P <0.000 1),the average number of IFNγ spots produced by spleen cells(t=9.235,P <0.000 1),and the number of CD4~+ and CD8~+T lymphocytes produced(t=10.870 and 6.200,P=0.008 4and 0.025 0,respectively) in mice of NA group increased significantly.The mice in PBS group all died within 5 days after infection,while in NP group,one mouse died 5 days after infection,and the other five mice survived.Conclusion The NP protein of influenza A virus expressed by Bac-to-Bac TOPO system has high immunogenicity and can produce strong humoral and cellular immune responses,which can provide certain protection for mice against influenza virus infection.

Objective: To express recombinant human soluble fibroblast growth factor receptor 1 ( sFGFR1) and study its antagonistic activity on FGF. Methods: Human sFGFR1 cDNA, isolated from human lung fibroblast cells with RT-PCR was confirmed by DNA sequencing and cloned into pYEX4T-1 yeast expression vector. The recombinant sFGFR1 was expressed in DY150 yeast cells and the product was identified by SDS-PAGE and Western blot. The activity of recombinant sFGFR1 was detected in N1H3T3 proliferation inhibition assay. Results: GSF-sFGFR1 fusion protein was expressed in yeast cells and was observed as a band of 60 Id) on a SDS- PAGE gel and by Western blot. The recombinant fusion protein was also found to be able to suppress FGF-induced proliferation of NIH3Th cells. Conclusion: Recombinant human GST-sFGFR1 fusion protein was expressed in yeast efficiently and showed natural biological activities.

This study was aimed to construct a clinical trial technological platform of traditional Chinese medicine (TCM) live disease in accordance with the international good clinical practice (GCP) standard. Through the con-struction of technological platforms such as clinical new medicine ethics committee of liver disease of TCM, ex-perimental methods, curative effect evaluation, data management, and etc., the clinical evaluation method and in-dex system of new drugs with the curative effect characteristics of liver disease of TCM were established. It was concluded that under the construction of relevant technological platform, an effect evaluation method and index system with TCM diagnosis and treatment characteristics were built . And a key clinical trial technological platform of new drugs of TCM liver disease was established to meet requirements of the international GCP standard.

Objective To analyze the characteristics of the Chinese Medical Equipment Journal's articles and authors as well as their spatial & temporal distribution,and then put forward some advices for the development of the journal.Methods 4 434 articles of the journal from 2002 to 2008 were sampled for bibliometrics analysis from the aspects of column,author,region,organization,publishing delay and foundation support.Results With an abundant manuscript source mostly from the developed areas such as Beijing,Guangdong Province,Shandong Province and Tianjin,the journal set its columns reasonably and flexibly,and thus gifted with a short publishing delay.The coauthor degree and cooperation rate were less than the journals in other fields,and the number of foundation-supported articles,also less than others,but was increasing yearly.Conclusion Chinese Medical Equipment Journal,with a high-quality manuscript source and great influence in the field,can be considered as a platform for communication and academic achievements publishing.

Objective To study the effect of angiotensin converting enzyme inhibitory peptides (ACEIP) on blood pressure of spontaneously hypertensive rats.Methods One hundred and four spontaneously hypertensive rats(SHR) were randomized to thirteen groups (n= 8,each group) and received one of following approaches:saline,captopril (10 mg/kg),low-dose (200 ?g/kg),medium-dose(400 ?g/kg) and high-dose (800 ?g/kg) of the three ACEIPs:VLPVP,KVLPVP,VLPVPR respectively.Thirty-two WKY rats were randomized to 4 groups(n=8 in each group),VLPVP,KVLPVP,VLPVPR (800 ?g/kg by gavage) and saline were administrated.The systolic blood pressure was measured before and after the administration of ACEIP.Results Compared with the control group,medium-dose and high-dose of VLPVP,KVLPVP and VLPVPR reduce blood pressure in hypertensive rats and the effect can last for above 6 hours [decline of blood pressure:high-dose:(40.9?15.7),(38.0?21.3) and (43.2?12.6) mmHg;medium-dose:(28.3?23.0),(24.7?21.5) and (26.2?9.9);control:(8.5?5.1),(6.2?4.1) and (7.1?5.5)mmHg;all P0.05].Conclusion Recombinant ACEIP acutely reduce high blood pressure in SHR,the antihypertensive effects persist for at least 6 hours,with no effect on normotensive WKY rats.

Anaplastic lymphoma kinase (ALK) rearrangement is one of the most potent carcinogenic genes in non-small cell lung cancer (NSCLC).The first-generation ALK inhibitor such as crizotinib is superior to chemotherapy for NSCLC patients with ALK rearrangement.At the same time,more and more studies have reported ALK inhibitors in brain metastases of NSCLC patients with intracranial efficiency.However,despite the initial clinical data of first-generation ALK inhibitors in the treatment of ALK-positive NSCLC with brain metastases,different degrees of recurrence of tumors after acquired resistance have posed new challenges for follow-up treatment of cancer patients.A new generation of ALK inhibitors,such as alectinib;ceritinib,AP26113 and PF-06463922 have emerged to solve this problem.

OBJECTIVE To investigate the expression of SleX and CD24 in nasal inverted papilloma and its pathologic features.METHODS HE staining were conducted to study the pathologic features in specimens of 11 cases with nasal inverted papilloma. Further,immunohistochemistry stain for SleX and CD24 were performed in total specimens.RESULTS One case(9.1%) was diagnosed as severe atypical hyperplasia but tumor cells did not invaded basal membranes.SleX staining located at cell membranes. Positive SleX staining was found in 9 specimens (81.8%) and 1 normal nasal epithelium (16.7%).CD24 staining located in cytoplasm.Positive CD24 staining was found in 8 specimens of nasal inverted papilloma (72.7%). CD24 was negative in nasal epithelial cells and only a few lymphocytes were positive.CONCLUSION Some cases of nasal inverted papilloma are diagnosed with severe atypical hyperplasia.Most of cases express CD24,so nasal inverted papilloma may be a borderline tumor.Expression of SleX and infiltration of inflammatory cells suggest that nasal inverted papilloma may be related to inflammatory reactions.

phological changes in rat liver tissues. TLR4 and NF-κB expression in the liver tissues were measured by im-munohistochemistry . The results showed that compared with the normal group , the serum levels of transaminase ( ALT , AST ) and endotoxin of the model group were higher ( P < 0 . 01 ); and the degree of liver pathology injury was significantly increased; the TLR4 and NF-кB expression were increased (P < 0.01). Compared with the model group, the serum levels of transaminase (ALT, AST) and endotoxin of the experimental group were lower (P < 0.01), the degree of liver pathology injury was significantly lighter; the TLR4 and NF-кB expression were significantly lower (P < 0.01). It was concluded that the WYJDHY granules has a good role in the prevention and treatment of liver injury of rat model of hepatic failure IETM through the downregulation of liver expression of TLR4 and NF-кB in rat liver tissues , reducing serum levels of endotoxin , which may be one of the mecha-nisms on hepatic failure treatment .

Objective To explore the expression of N-myc downstream regulated gene 1 (NDRG1) in gastric carcinoma and the relationship with clinicopathological parameters and prognosis. Methods The expression of NDRGI was detected by immunohisto chemistry in formalin-fixed and paraffin-embedded sections with a total of 220 specimens including 110 gastric carcinoma and 110 corresponding paraneoplastic tissue. The correlation between clinicopathological parameters and the expression of NDRG1 in gastric carcinoma were also analyzed. Results Low expression of NDRG1 was detected in most gastric carcinoma sections. Among the gastric cancer tissues, NDRG1 protein expression was significantly lower in tumors with more advanced pathological stage, local tumor invasion and lymphatic metastases. There was no significant difference in sex, age, tumor differentiation and gross types of the tumor. The 1-, 3- and 5 year survival and disease free survival in patients with low NDRG1 protein expression was 84.2%, 53.9%, 21.1%, and 60.5%, 31.6%, 19.7%, respectively, which was signifivantly poorer when compared with patients with high NDRG1 protein expression. Conclusion The expression of NDRG1 is low in the majority of patients with gastric carcinoma, which was in a close relationship with advanced stage, local invasion and lymphatic metastases of gastric carcinoma. NDRG1 may be a candidate metastasis suppressor gene.

Background and purpose:A pressing obstacle in clinical chemotherapy is drug resistance in breast cancer.A nano-delivery system,which has many advantages as a drug carrier,such as carrying anticancer drugs,can be used effectively to overcome drug resistance in tumors.This paper examined a new nano-delivery system,called calcium phosphate and glycerophosphocholine-mPEG(CAP/GPC-MPEG)composite nanoparticle and its influence on the cellular drug uptake of BCRP-over expressing mitoxantrone(MIT)-resistant breast cancer cell MCF-7/MIT.This paper will also examine its effect on overcoming drug resistance in the MCF-7/MIT cells.Methods:After the calcium phosphate and GPC-mPEG composite nanoparticles were designed and prepared,the entrapment efficiency and in vitro drug release of mitoxantrone-loaded nanoparticles were investigated.Quantitative comparisons were made between cellular uptake of drug-loaded nanoparticles and free drugs.Finally,a confocal laser scanning microscopy Was used to compare the subcellular distribution of drug-loaded nanoparticles and the free drugs.Results:Calcium phosphate and GPC-mPEG composite nanoparticles were nanoporous spherical particles with diameters between 50-100 mn.The MIT-loaded nanoparticles have an entrapment efficiency of(89.45±0.05)%.Although the drug-loaded nanoparticles showed an initial burst of drug release,it was followed by a more sustained release.The concentration of mitoxantrone was 1.89 times treated with MIT-loaded nanoparticles for 1 h compared to that treated with free mitoxantrone for 1 h in MCF-7/MIT cells.and which was 2.33 times in MCF-7 cells.Fluorescent red mitoxantrone appeared in the cytoplasm and nucleus of the MCF-7 and MCF-7,MIT cells treated with MlT-loaded nanoparticles whereas it is almost undetected in both cells treated with free mitoxantrone.Conclusion:Calcium phosphate and GPC-mPEG composite nanoparticles Can promote the cellular uptake and entering of mitoxantrone to the nucleus in MCF-7 and its corresponding BCRP-over expressing MIT-resistant MCF-7/MIT breast cancer cell lines.This nanoparticle is a potential nano-carrier for overcoming drug resistance in tumors.