1.The research of teaching supervision mechanism in modern military academy
Lin OUYANG ; Yungui WANG ; Jianjun HUANG ; Jidong HUANG ; Hongxiao FAN
Chinese Journal of Medical Education Research 2017;16(3):235-238
In this study,with the mechanism for teaching supervision in military colleges and universities construction as the starting point,the author makes an in-depth analysis of the problems existing in the current teaching supervision system of military academies,such as the imperfect mechanism,the limitation of the function,the lack of full-time staff,the lag of information technology and so on,and investigates and surveys the status quo of teaching supervision at home and abroad.It also puts forward some countermeasures,such as constructing the comprehensive supervision system,realizing the function transformation,promoting the innovation of the mode,expanding the connotation of the supervision,optimizing the structure of the team,developing the professional training,and strengthening the information construction and so on.
2.Cloning,expression and immuno-protection analysis of a gene encoding tro-ponin T of Schistosoma japonicum(SjTnT)
Xinzhuo WANG ; Yang HONG ; Hongxiao HAN ; Changjian LI ; Xiaodan CAO ; Ke LU ; Hao LI ; Chuangang ZHU ; Zhiqiang FU ; Jiaojiao LIN
Chinese Journal of Schistosomiasis Control 2014;(4):394-398
Objective To clone cDNA encoding troponin T of Schistosoma japonicum(SjTnT),and evaluate the protective efficacy induced by recombinant SjTnT in BALB/c mice against S. japonicum challenge infection. Methods The SjTnT gene was amplified from 28-day-schistosome cDNAs by PCR and then subcloned into pET28a(+). The recombinant SjTnT protein (rSjTnT)was expressed in Escherichia coli BL21(DE3)cells. The serum specific to rSjTnT was prepared by immunized BALB/c mice with the recombinant antigen,and the immunogenicity of rSjTnT was detected by Western blotting and ELISA. The immuno-protective efficacy induced by rSjTnT in BALB/c mice was evaluated according to the reduction in worm and egg counts. Results The cDNA encoding SjTnT was successfully cloned and expressed in E. coli. Western blotting showed that rSjTnT had a good immunogenicity. The high level of specific IgG antibodies was detected,and 33.89% worm reduction and 43.94% liver egg reduction were obtained in mice vaccinated with rSjTnT combined with Seppic 206 adjuvant compared with those in the adjuvant control group. Conclusions rSjTnT could induce partial immuno-protection against S. japonicum infec-tion in BALB/c mice. This study provided a basic for understanding the biological function of SjTnT.
3.Relationship between changes of serum IL-34, RF and RANKL levels in patients with rheumatoid arthritis and osteoporosis
International Journal of Laboratory Medicine 2017;38(18):2540-2542
Objective To investigate and analyze the changes of serum interleukin-34(IL-34),rheumatoid factor(RF) and nuclear factor κB receptor activator ligand(RANKL) in patients with rheumatoid arthritis Its relationship with osteoporosis.Methods A total of 40 patients suffering from rheumatoid arthritis in the hospital between January 2016 and January 2017 were included in the study group,and 40 healthy subjects who underwent the same period of physical examination at the same time were enrolled as controls.By enzyme-linked immunosorbent assay(ELISA) were detected in 2 groups of serum IL-34 and RANKL levels were detected by latex agglutination assay of serum RF levels in 2 groups,and routine testing in patients with bone mineral density,erythrocyte sedimentation rate(ESR),C reactive protein(CRP) and anti CCP antibody.And the modified DAS28 score of disease activity of rheumatoid arthritis evaluation.Results The levels of serum IL-34,RF and RANKL in the study group were significantly higher than those in the control group(P<0.05).The incidence of osteoporosis in the study group was significantly higher than that in the control group(P<0.05).The levels of IL-34,RF and RANKL in the osteoporosis patients were significantly higher than those in the normal group(P<0.05).The Spearman correlation analysis showed that the serum IL-34 levels were positively correlated with ESR,CRP,anti-CCP and DAS28 (P< 0.05).There was a negative correlation between serum IL-34,RF and RANKL levels and bone mineral density(P<0.05).Conclusion The levels of serum IL-34,RF and RANKL in patients with rheumatoid arthritis were significantly increased,and the above indexes were negatively correlated with bone mineral density.It suggested that IL-34 may be involved in osteoporosis.
4.Cloning, expression and protective efficacy evaluation of radiation sensitive protein 23 (RAD23) from Schistosoma japonicum.
Changjian LI ; Min ZHANG ; Yang HONG ; Yanhui HAN ; Xiaodan CAO ; Hongxiao HAN ; Zhiqiang FU ; Chuangang ZHU ; Ke LU ; Hao LI ; Jiaojiao LIN
Chinese Journal of Biotechnology 2014;30(11):1669-1678
Radiation sensitive protein 23 (RAD23) is a nucleotide excision repair (NER) protein that plays an important role in Ubiquitin-proteasome pathway (UPP). Schistosoma japonicum radiation sensitive protein23 (SjRAD23) cDNA sequences were amplified by PCR and cloned into pET28a (+) vector to construct recombinant expression plasmid pET28a(+)-SjRAD23. The recombinant protein was expressed as both inclusion bodies and the supernatant in Escherichia coli BL21 (DE3) cell. Immunofluorescence observation shows that SjRAD23 was mainly distributed on the tegument surface of the worms. ELISA assay reveals that specific IgG, IgG1 and IgG2a antibodies could be detected in the sera of rSjRAD23 immunized mice. Western blotting analysis shows that the recombinant SjRAD23 could be recognized by serum specific to soluble adult worm antigen of S. japonicum. BALB/c mice vaccinated with rSjRAD23 combined with 206 adjuvant revealed 35.94% worm reduction and 40.59% liver egg reduction when compared with that of the adjuvant control
Animals
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Antibodies, Helminth
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blood
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Blotting, Western
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Cloning, Molecular
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DNA Repair Enzymes
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genetics
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metabolism
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DNA, Complementary
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli
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Genetic Vectors
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Helminth Proteins
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genetics
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immunology
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Immunoglobulin G
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blood
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Mice
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Mice, Inbred BALB C
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Recombinant Proteins
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genetics
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immunology
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Schistosoma japonicum
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genetics
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metabolism
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Schistosomiasis japonica
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prevention & control
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Vaccines
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immunology
5.Prognostic value of FDG PET-CT metabolic parameters before radiotherapy in patients with esophageal cancer receiving radiotherapy
Hongxiao LIN ; Liting QIAN ; Bo PAN ; Jianhua YAN
Chinese Journal of Radiation Oncology 2017;26(12):1394-1398
Objective To investigate the prognostic value of metabolic parameters of 18F-fluorodeoxyglucose positron emission tomography/computed tomography(FDG PET-CT)before radiotherapy in patients with esophageal cancer receiving radiotherapy. Methods We retrospectively analyzed the clinical data of 37 patients with stage Ⅱ-Ⅲ esophageal squamous cell carcinoma admitted to Anhui Provincial Hospital from 2010 to 2015,who underwent PET-CT scans within 4 weeks before radiotherapy. The following information was collected:age,sex,the location of primary lesion,clinical stage,and treatment regimen.The maximum standard uptake value(SUVmax), mean standard uptake value(SUVmean), metabolic tumor volume(MTV), PET tumor length(PTL),and total lesion glycolysis(TLG)were calculated. The optimal cut-off values of SUVmax, SUVmean,MTV,PTL,and TLG were determined by the receiver operating characteristic(ROC)curve. The Kaplan-Meier method was used to calculate survival rates,and the log-rank test was used for survival comparison and univariate prognostic analysis;the Cox model was used for multivariate prognostic analysis of significant variables. Results The area under the ROC curve suggested that MTV>TLG>PTL=0.713>0.679>0.670. The Kaplan-Meier survival analysis showed that patients had better survival time when MTV was less than 27.44 cm3,TLG was less than 166.2 g,and PTL was less than 36.74 mm(P<0.05).The Cox multivariate regression analysis showed that only MTV was an independent prognostic factor for radiotherapy in patients with esophageal cancer(P=0.000). Conclusions MTV, TLG and PTL have certain values in predicting the prognosis of patients with esophageal cancer before radiotherapy, and are helpful for selecting and adjust the clinical treatment regimen and improving the survival rate and prognosis.
6.Cloning, expression and characterization of a gene encoding alpha2 subunit of the proteasome in Schistosoma japonicum.
Yang HONG ; Hongxiao HAN ; Jinbiao PENG ; Ye LI ; Yaojun SHI ; Zhiqiang FU ; Jinming LIU ; Xiangrui LI ; Jiaojiao LIN
Chinese Journal of Biotechnology 2010;26(4):509-516
The 26S proteasome is a proteolytic complex responsible for the degradation of the vast majority of eukaryotic proteins. Regulated proteolysis by the proteasome is thought to influence cell cycle progression, transcriptional control, and other critical cellular processes. A novel Schistosoma japonicum gene (GenBank Accession No. AY813725) proteasome alpha2 subunit (SjPSMA2) was cloned. Sequence analysis revealed that the ORF of SjPSMA2 gene contains 708 nucleotides encoding 235 amino acids, and the molecular weight was estimated to be 25.84 kDa. Real-time PCR analysis showed that this gene expressed in 7 d, 13 d, 18 d, 23 d, 32 d and 42 d schistosoma. The mRNA level of SjPSMA2 was lower in 7 d and 23 d schistosomulum than that in other stages. The SjPSMA2 cDNA fragment was subcloned into an expression vector pET28a(+) and transformed into E. coli BL21 (DE3) cells. After induction with IPTCQ the 30 kDa fusion protein was produced as included bodies. Western-blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation, and the protein in native could be detected. After immunization of BALB/c mice with the fusion protein, the reduction rates of worm counts and liver egg counts were 12.33% and 35.23%. ELISA results revealed that the vaccinated group showed a significant increase in the level of IgG antibody. This study provided an important basis for investigating the regulation mechanism of the proteasome during the development of Schistosoma japonicum.
Animals
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Antibodies, Helminth
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blood
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Base Sequence
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Genes, Helminth
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Helminth Proteins
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genetics
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metabolism
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Immunization
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Liver
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parasitology
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Male
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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Parasite Egg Count
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Proteasome Endopeptidase Complex
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biosynthesis
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genetics
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immunology
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Rabbits
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Recombinant Proteins
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biosynthesis
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genetics
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immunology
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Schistosoma japonicum
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genetics
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metabolism
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Vaccines, Synthetic
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immunology
7.Cloning and expressing of cyclophilin B gene from Schistosoma japonnicum and the analysis of immunoprotective effect.
Jinbiao PENG ; Hongxiao HAN ; Yang HONG ; Yan WANG ; Fanji GUO ; Yaojun SHI ; Zhiqiang FU ; Jinming LIU ; Guofeng CHENG ; Jiaojiao LIN
Chinese Journal of Biotechnology 2010;26(3):317-323
The present study was intend to clone and express the cDNA encoding Cyclophilin B (CyPB) of Schistosoma japonicum, its preliminary biological function and further immunoprotective effect against schistosome infection in mice. RT-PCR technique was applied to amplify a full-length cDNA encoding protein Cyclophilin B (Sj CyPB) from schistosomula cDNA. The expression profiles of Sj CyPB were determined by Real-time PCR using the template cDNAs isolated from 7, 13, 18, 23, 32 and 42 days parasites. The cDNA containing the Open Reading Frame of CyPB was then subcloned into a pGEX-6P-1 vector and transformed into competent Escherichia coli BL21 for expressing. The recombinant protein was renaturated, purified and its antigenicity were detected by Western blotting, and the immunoprotective effect induced by recombinant Sj CyPB was evaluated in Balb/C mice. The cDNA containing the ORF of Sj CyPB was cloned with the length of 672 base pairs, encoding 223 amino acids. Real-time PCR analysis revealed that the gene had the highest expression in 18-day schistosomula, suggesting that Sj CyPB was schistosomula differentially expressed gene. The recombinant protein showed a good antigenicity detected by Western blotting. Animal experiment indicated that the vaccination of recombinant CyPB protein in mice led to 31.5% worm and 41.01% liver egg burden reduction, respectively, compared with those of the control. A full-length cDNA differentially expressed in schistosomula was obtained. The recombinant Sj CyPB protein could induce partial protection against schistosome infection.
Animals
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Antigens, Helminth
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immunology
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Cloning, Molecular
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Cyclophilins
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biosynthesis
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genetics
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immunology
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DNA, Complementary
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genetics
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Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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genetics
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Immunization
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Mice
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Mice, Inbred BALB C
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Recombinant Proteins
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biosynthesis
;
genetics
;
immunology
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Schistosoma japonicum
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genetics
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immunology
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Schistosomiasis japonica
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prevention & control
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Vaccines, Synthetic
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biosynthesis
;
immunology
8.Cloning, eukaryotic expressing and function analysis of Schistosoma japonicum apoptosis gene Sjcaspase3.
Tao WANG ; Yang HONG ; Hongxiao HAN ; Chao LV ; Bingguang JIA ; Xiaodan CAO ; Qian HAN ; Ke LU ; Hao LI ; Zhiqiang FU ; Jiaojiao LIN
Chinese Journal of Biotechnology 2016;32(7):889-900
For further research of the apoptosis mechanism of Schistosoma japonicum (S. japonicum). The cDNA encoding Sjcaspase3 of Schistosoma japonicum was amplified by polymerase chain reaction (PCR) technique, which contained 900 nucleotides and encoded 299 amino acids. The theory molecular weight and isoelectric point (PI) of the deduced protein is 33.5 kDa and 6.39, respectively. Real-time PCR was used to analyze the transcription profiles of Sjcaspase3 at different development stages of S. japonicum. The results showed that this gene was expressed in all stages of S. japonicum with the highest expression in 21d worms, and the level of gene transcription in 42 d female worms was higher than that of male worms. The recombinant plasmid pXJ40-FLAG-Sjcaspase3 was constructed and transfection into Hela cells successfully. Real-time PCR and Western blotting analysis showed Sjcaspase3 was successfully expressed in Hela cells. Enzyme activity analysis revealed that recombinant Sjcaspase3 possessed the activity to cut substrate DEVD. Flow cytometry proved that Sjcaspase3 could induce early apoptosis of Hela cells. The results provide the basis for proceeding further study on the biological function of Sjcaspase3 and better understand the apoptosis mechanism of S. japonicum.
Animals
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Apoptosis
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Blotting, Western
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Caspase 3
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genetics
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metabolism
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Cloning, Molecular
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DNA, Complementary
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Female
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HeLa Cells
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Helminth Proteins
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genetics
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metabolism
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Humans
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Male
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Real-Time Polymerase Chain Reaction
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Recombinant Proteins
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Schistosoma japonicum