Objective To explore the effect of let-7c-1 on the learning and memory of PTZ-induced epileptic rats and its relevant mechanism.Methods A model of temporal lobe epilepsy (TLE) was induced via PTZ kindling in SD male rats.The epileptic rats were divided into epilepsy group,agomir-control group,let-7c-1 agomir group (12 rats for each).Twelve rats were served as a negative control group.The behavior and the expression levesl of let-7c-1,Bcl-2 protein and Caspase3 were evaluated at 28 days following PTZ.Results Compared to the negative group,the escape latency of epilepsy group was prolonged and the crossing times as well as the quadrant total distance in the target were reduced (P＜0.05).However,those parameters were not significantly different between the epilepsy group and the agmoir-control group (P＞0.05).Compared to the agmoir-control group,the escape latency of let-7c-1 agomir group was prolonged and the crossing times as well as the quadrant total distance in the target were reduced (P＜ 0.05).The expression levels of let-7c-1 and let-7c-1 were 1.35±0.32 in agmoir-control group and 62.53±21.01 in agomir group (F=50.97,P＜0.05).The expression levels of let-7c-1 were higher in let-7c-1 agomir group than in other groups (P＜0.05).Compared to the negative group,the expressions of Bcl-2 protein in other groups were decreased (P＜0.05) and the Caspase3 protein were increased (P＜0.05).Compared to the agomir-control group,the expression of Bcl-2 protein was significantly decreased and the expression of Caspase3 protein was significantly increased in let-7c-1 agomir group (P＜0.05).Conclusions The present study shows that let-7c-1 may impair the learning and memory of PTZ-induced epileptic rats through decreasing the Bcl-2 protein and increasing Caspase3 protein in the hippocampus.

Objective To investigate the pollution condition of Salmonella in food with phage. Methods Salmonella in 413 samples of food were detected by the diagnosis of Salmonella phage with biochemical and serological identification. Results 119 Salmonella were detected in 99 positive samples and the isolating rate was 24%. Conclusion The disservice of Salmonella is mainly from the meat food and eggs. The detection method of phage is fast,convenient and reliable.

Radiation-induced fibrosis, especially radiation-induced muscle fibrosis (RIMF), is a common complication associated with radiotherapy. After radiotherapy, there is a gradual worsening of muscle stiffness and joint mobility impairments over time, which severely affects the quality of life of cancer patients. This article aims to review the latest research progress on the mechanisms and diagnosis and assessment methods of RIMF, as well as its rehabilitation strategies. This review provides systematic and effective health management and rehabilitation strategies to improve the prognosis of patients with RIMF.

OBJECTIVETo investigate the roles of Domain I and Domain II of hepatitis C virus (HCV) 5' untranslated region (UTR) in the translation initiation activity of HCV 5'UTR in different host cell lines.

METHODSThe eukaryotic expression plasmid pCMVNCRLuc (pCN1), in which full-length HCV 5'UTR regulates firefly luciferase expression, was modified by deleting Domain I and the downstream single-stranded sequence (43 bp in total) from the UTR (pCNl-d2), Domain I with the downstream single-stranded sequence and Domain II (118 bp in total) from the UTR (pCNl-d3), or the total UTR (pCNl-d5). The modified plasmids were transfected via liposome into different cell lines with pRL-TK plasmid co-transfected as the normalization control. At 36 h after the transfection, the total cellular RNA was harvested for semi-quantitative RT-PCR, and the relative expression activities of luciferase were assayed with a dual luciferase reporter gene assay system. The translation initiation activities of the truncated HCV 5'UTRs in different translation systems were analyzed.

RESULTSDeletion of Domain I and the downstream single-stranded sequence caused no significant changes of the translational activity of HCV 5'UTR in Hela or C6 cells, but decreased the translational activity by 46% in L-02 cells and increased the translational activity by 46% in 293T cells. Deletion of both Domain I and Domain II resulted in decreased translational activity of HCV 5'UTR by 51% in HeLa cells, but increased the translational activity by 40% in L-02 cells, 60% in C6 cells and 135% in 293T cells.

CONCLUSIONSDomain I and Domain II of HCV 5'UTR perform cell type-specific roles in HCV IRES-driven translation initiation.

5' Untranslated Regions ; Genes, Reporter ; HeLa Cells ; Hepacivirus ; genetics ; Humans ; Luciferases ; Plasmids ; Protein Biosynthesis ; genetics ; RNA, Viral ; genetics ; Transfection

Objective To investigate the roles of Domain I and Domain II of hepatitis C virus (HCV) 5' untranslated region (UTR) in the translation initiation activity of HCV 5'UTR in different host cell lines. Methods The eukaryotic expression plasmid pCMVNCRLuc (pCN1), in which full-length HCV 5'UTR regulates firefly luciferase expression, was modified by deleting Domain I and the downstream single-stranded sequence (43 bp in total) from the UTR (pCNl-d2), Domain I with the downstream single-stranded sequence and Domain II (118 bp in total) from the UTR (pCNl-d3), or the total UTR (pCNl-d5). The modified plasmids were transfected via liposome into different cell lines with pRL-TK plasmid co-transfected as the normalization control. At 36 h after the transfection, the total cellular RNA was harvested for semi-quantitative RT-PCR, and the relative expression activities of luciferase were assayed with a dual luciferase reporter gene assay system. The translation initiation activities of the truncated HCV 5'UTRs in different translation systems were analyzed. Results Deletion of Domain I and the downstream single-stranded sequence caused no significant changes of the translational activity of HCV 5'UTR in Hela or C6 cells, but decreased the translational activity by 46%in L-02 cells and increased the translational activity by 46%in 293T cells. Deletion of both Domain I and Domain II resulted in decreased translational activity of HCV 5'UTR by 51%in HeLa cells, but increased the translational activity by 40%in L-02 cells, 60%in C6 cells and 135%in 293T cells. Conclusions Domain I and Domain II of HCV 5'UTR perform cell type-specific roles in HCV IRES-driven translation initiation.

Objective To investigate the roles of Domain I and Domain II of hepatitis C virus (HCV) 5' untranslated region (UTR) in the translation initiation activity of HCV 5'UTR in different host cell lines. Methods The eukaryotic expression plasmid pCMVNCRLuc (pCN1), in which full-length HCV 5'UTR regulates firefly luciferase expression, was modified by deleting Domain I and the downstream single-stranded sequence (43 bp in total) from the UTR (pCNl-d2), Domain I with the downstream single-stranded sequence and Domain II (118 bp in total) from the UTR (pCNl-d3), or the total UTR (pCNl-d5). The modified plasmids were transfected via liposome into different cell lines with pRL-TK plasmid co-transfected as the normalization control. At 36 h after the transfection, the total cellular RNA was harvested for semi-quantitative RT-PCR, and the relative expression activities of luciferase were assayed with a dual luciferase reporter gene assay system. The translation initiation activities of the truncated HCV 5'UTRs in different translation systems were analyzed. Results Deletion of Domain I and the downstream single-stranded sequence caused no significant changes of the translational activity of HCV 5'UTR in Hela or C6 cells, but decreased the translational activity by 46%in L-02 cells and increased the translational activity by 46%in 293T cells. Deletion of both Domain I and Domain II resulted in decreased translational activity of HCV 5'UTR by 51%in HeLa cells, but increased the translational activity by 40%in L-02 cells, 60%in C6 cells and 135%in 293T cells. Conclusions Domain I and Domain II of HCV 5'UTR perform cell type-specific roles in HCV IRES-driven translation initiation.

Objective:To investigate the effects of applying the "4C" teaching method combined with the Gibbs' reflective cycle to clinical nursing teaching in the department of urology.Methods:A total of 68 nursing students who practiced in the urology department of our hospital were divided using a lottery method into experimental group ( n=35, entering the second and third wards for internship) and control group ( n=33, entering the first and fourth wards). The experimental group was taught using the "4C" teaching method combined with the Gibbs' reflective cycle, including four teaching steps (connection, concept, concrete practice, and conclusion) and regular reflective discussions. The control group received traditional teaching, which covered entrance education, centralized theoretical training, skills demonstration, and exit assessment. At the end of internship, the two groups were compared in terms of self-directed learning ability, nursing competency, and theoretical and practical scores. SPSS 23.0 was used to perform the t test. Results:The self-directed learning scores of the experimental group and the control group were (227.37±12.91) and (207.09±16.27), respectively. The nursing competency scores were (156.66±12.49) and (138.06±17.23), respectively. The experimental group was significantly superior to the control group in self-directed learning ability ( t=5.71, P<0.001), nursing competency ( t=5.12, P<0.001), theoretical score ( t=3.03, P=0.004), and practical score ( t=4.88, P<0.001). Conclusions:The "4C" teaching method combined with the Gibbs' reflective cycle can effectively improve nursing students' self-directed learning ability and nursing competency, and help them better master knowledge and skills.

Objective: To evaluate an assay permitting amplification of target 5′-untranslated region (5′-UTR) sequences directly from clinical specimens and distinction among serotypes of enterovirus (EV). Methods: A total of 518 rectal swabs and 148 nasal swabs tested positive by pan-enterovirus real-time PCR were collected. 5′-UTR and the viral protein 1 (VP1) gene fragments were amplified and sequenced separately for serotyping. The inconsistent samples by 5′-UTR and VP1 serotyping were further determined by using the serotype-specific RT-PCR. Results: A total of 553 (83.0%) samples were detected by 5′-UTR serotyping and 318 (47.7%) were detected by VP1 serotyping in all 666 positive specimens, and there was significant difference in the detection rates between two methods in rectal and nasal swabs (P<0.001). For the rectal swabs, the mainly detected serotypes were CoxA6 (217), CoxA16 (88), EVA71 (40), CoxA10 (28) and CoxA4 (27) by 5′-UTR serotyping. Compared with the VP1 serotyping, the sensitivity and specificity of 5′-UTR serotyping were 57.1%-100% and 67.4%-98.1% respectively, with varied consistence with serotypes (kappa value 0.214-0.283). For the nasal swabs, the most frequently detected serotype was EVD68, with the sensitivity of 100%, the specificity of 91.1%, and the poor consistence (kappa value 0.217). CoxA6, CoxA16, EVA71, CoxA10 and EVD68 were further confirmed by serotype-specific RT-PCR. Using VP1 serotyping combined with serotype-specific RT-PCR as a reference method , the effect of performance of 5′-UTR serotyping on diagnosis was increased. Conclusions The performances of 5′-UTR serotyping in enterovirus vary with serotypes. The application of 5′-UTR serotyping should be considered comprehensively according to the purpose of the study.

To construct the recombinant vector pBV220-scFv and express anti-clenbuterol (CBL) scFv antibody in Escherichia coli, we amplified the scFv gene using plasmid pCANTABSE-CBL as a template, recombined it with pPICZalphaA, then amplified the scFv-His-tag gene from plasmid pPICZalphaA-scFv and linked it with expression plasmid pBV220. We identified the recombinant plasmid by restrictive enzyme digestion, PCR amplification and sequence analysis. Finally, we transformed the recombinant vector into E. coli DH5alpha that was temperature-induced and expressed recombinant protein. We identified the recombinant protein by SDS-PAGE, Western blotting and indirect competitive ELISA. The results show that recombinant plasmid pBV220-scFv contained the inserted fragment with highest homology about 99.8%. The expression of scFv induced by temperature show 37 kD Mw and anti-His-tag mAb recognized-activity by SDS-PAGE and Western blotting respectively, and could competitively combine with CBL, the IC50 is 4.55 ng/mL. The recombinant plasmid pBV220-scFv is constructed and expresses the scFv gene of CBL in E. coli successfully. This study suggests the corresponding immunoassay methods could be set up by the recombinant scFv.
Antibodies ; immunology ; Clenbuterol ; immunology ; Cloning, Molecular ; Genetic Vectors ; genetics ; Immunoglobulin Fragments ; biosynthesis ; genetics ; immunology ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

【Objective】 To observe the clinical efficacy of ultrasound-guided autologous platelet- rich plasma (PRP) injection for the treatment of common peroneal nerve compression syndrome, and to provide reference for the treatment of peripheral nerve compression diseases. 【Methods】 A patient with common peroneal nerve compression syndrome was treated with ultrasound-guided injection of autologous PRP for nerve water separation. The efficacy of PRP injection treatment on common peroneal nerve compression syndrome was evaluated by observing the patient′s symptoms, electrophysiological and imaging manifestations. 【Results】 After systematic injection therapy, the patient′s symptoms, signs and electromyography showed significant improvement, and there were no significant adverse reactions after 3 months of follow-up. 【Conclusion】 Ultrasound guided PRP injection therapy improves neurological dysfunction in common peroneal nerve compression syndrome.