Research progress of avermectins in anti-tumor / 国际肿瘤学杂志

Avermectins can affect biological processes of multiple tumor, including tumor cell proliferation and metastasis, cell cycle arrest, induction of apoptosis and autophagy, regulation of tumor microenvironment and tumor stem cells. Avermectins can be administered alone or combined with chemotherapeutic drugs to reverse multidrug resistance. To further explore the anti-tumor mechanism of avermectins will provide reliable experimental and theoretical guidance for future clinical application.

The best evidence summary for prevention of hypothermia at birth in newborn / 中国实用护理杂志

Objective:To retrieve and summarize evidence for prevention of hypothermia at birth in newborn.Methods:Databases such as Up To Date, BMJ Best Practice, National Institute for Health and Care Excellence(NICE), Joanna Briggs Institute (JBI), Cochrane Library, Registered Nurses′Association of Ontario (RNAO), American Heart Association(AHA), Web of Science, PubMed, Chinese Biology Medical Literature database, Wanfang Med Online were searched to collect relevant evidence for prevention of hypothermia at birth in newborn, including guidelines, systematic reviews, evidence summaries and expert consensus. Two researchers independently evaluated the quality of the literature and extracted the data of the literature which met the criteria.Results:Six articles were selected, including 1 clinical decision support system, 2 guidelines, 1 systematic review and 2 expert consensuses. Nineteen pieces of best evidence were summarized.Conclusions:This study summarized the best evidence for the prevention of hypothermia at birth in newborn, and provided evidence-based support for clinical practice.

Liver histological changes after adefovir treating in chronic hepatitis B patients with HBeAg negative / 中国基层医药

Objective To investigate hepatic pathohistological changes(including pathology,HBV markers in liver tissue)in patients with KBeAg negative chronic hepatitis B after adefovir therapy for 1 year.Methods 35 patients with HBeAg negative chronic hepatitis B were administered with 10mg adefovir once a day for one year.The needle biopsy of liver were performed before and after treatment.The routine HE stained liver tissue sections were evaluated and Knodell pathological score were done.HBsAg and HBeAg in liver tissue were examined by immunohisochemisty method.Results After 1 year,18,17 and 11 cases had a significant reduction of their total hepatic HAI score,necroinflammation and fibrosis score.The immunohistochemistry examination showed HBeAg in liver decreased significantly,but HBsAg had no obvious alteration.Conclusion Significant improvement in both necroin flammation and fibrosis can be obtained in the majority of patients treated with adefovir for 1 year.More significant improvement in live histology can be obtained after extended treatment.

Construction and Expression of Bivalent Membrane-anchored DNA Vaccine Encoding Sj14FABP and Sj26GST Genes / 华中科技大学学报（医学）（英德文版）

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sjl4 or pVAC-Sj26 only to get two gene fragments including Sjl4 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES,resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sj14 and Sj26genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj 14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

Construction and expression of bivalent membrane-anchored DNA vaccine encoding Sjl4FABP and Sj26GST genes / 华中科技大学学报（医学）（英德文版）

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

Expression and purification of SARS coronavirus membrane protein / 华中科技大学学报（医学）（英德文版）

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-beta-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.

Expression and purification of SARS coronavirus membrane protein / 华中科技大学学报（医学）（英德文版）

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-beta-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.

Construction, expression and identification of a recombinant BCG vaccine encoding human Mycobacterium tuberculosis heat shock protein 65 / 华中科技大学学报（医学）（英德文版）

Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69% in total bacterial protein and 74.09% in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.