OBJECTIVE To explore study a method for rapid detection of bacterial infection in clinic to diagnose septicemia early.METHODS 16S rRNA gene of ten bacterial species was amplified with PCR,by using human genome DNA,HBV-DNA and Candida albicans as comparison.The sensitivity test was done by the method of gradual dilution of Escherichia coli.RESULTS The bacterial species were amplified and the products were 371 bp,but human genome DNA,HBV-DNA and C.albicans showed no amplification products.Sensitivity test showed that it could detect as low as 1.5?104/L of E.coli.CONCLUSIONS The method is rapid and highly specific and sensitive in detecting the existence of bacterial 16S rRNA gene.

OBJECTIVE To set up a quick method to detect pathogens in cerebrospinal fluid.METHODS The method of polymerase chain reaction(PCR),using a pair of universal primers targeted at the 16S rRNA gene,was adopted to amplify the DNA of bacterium.138 clinical specimens obtained from patients were examined by PCR method and bacterial culture method.RESULTS The positive rate was 39.86% of PCR and 17.39% of culture,the outcomes had statistical significance(P

OBJECTIVETo generate a candidate HPV16 vaccine for experimental and therapeutical use for cervical cancer.

METHODSThe mutants of HPV16 early E6 and E7 genes were inserted into a vaccinia virus expression vector. A strain of recombinant vaccinia virus was constructed through homologous recombination.

RESULTSShowed that the mutant E6 and E7 genes were located at TK gene region of vaccinia virus Tiantan strain in a head to head orientation under the control of early/late promoters, H6 and 7.5K respectively. Studies in mice indicated that VmE6E7 could elicit specific antibodies against E6 and E7, and retarded or prevented tumor development in a proportion of C57 BL/6 mice challenged by syngeneic HPV16E6 and E7 transformed tumor cells.

CONCLUSIONSThe success in constructing VmE6E7 provides a basis for the further development of HPV16 therapeutic vaccine.

Animals ; Female ; Genes, Viral ; genetics ; Genetic Vectors ; genetics ; immunology ; Mice ; Mice, Inbred C57BL ; Mutation ; Neoplasms, Experimental ; prevention & control ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Recombination, Genetic ; Repressor Proteins ; Transfection ; Vaccinia virus ; genetics ; immunology

OBJECTIVETo generate an HPV16 prophylactic vaccine candidate for cervical cancer.

METHODSHPV16 major capsid protein L1 gene and minor capsid protein L2 gene were amplified using PCR. These genes were mutated by PCR site-directed mutagenesis for removal of sequence motifs (TTTTTNT) which would cause transcription termination when expressed from a vaccinia virus early promoter, then inserted into a vaccinia virus expression vector. A strain replication-deficient recombinant vaccinia virus containing the mutant sequences was obtained through a homologous recombination and identified.

RESULTSThe nucleotide sequence remained the correct amino acid sequence of the L1 and L2 proteins after mutated. Full-length L1 and L2 proteins were generated in cells infected with the recombinant virus. The virus strain propagated at very low titer or could not reproduce in some kinds of cell derived from different human tissues.

CONCLUSIONSThe authors have generated a strain replication-deficient recombinant vaccinia virus expressing HPV16 L1 plus L2 proteins as an HPV16 prophylactic vaccine candidate for cervical cancer.

Capsid ; Capsid Proteins ; genetics ; Cell Line ; Cloning, Molecular ; Female ; Gene Expression ; Genetic Vectors ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; prevention & control ; Transfection ; Tumor Virus Infections ; prevention & control ; Uterine Cervical Neoplasms ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication

Tetramethylpyrazine (TMP), an effective component of traditional Chinese medicine Chuanxiong, is commonly used to resolve embolism. Its possible therapeutic effect against atherosclerosis has received considerable attention recently. Angiotensin II (Ang II) is highly implicated in the proliferation of vascular smooth muscle cells (VSMCs), resulting in atherosclerosis. The mechanisms of TMP in the proliferation of VSMCs induced by Ang II remain to be defined. The present study was aimed to study the effect of TMP on Ang II-induced VSMC proliferation through detection of nuclear factor-kappaB (NF-kappaB) activity and bone morphogenetic protein-2 (BMP-2) expression. Primary cultured rat aortic smooth muscle cells were divided into the control group, Ang II group, Ang II + TMP group and TMP group. Cells in each group were harvested at different time points (15, 30 and 60 min for detection of NF-kappaB activity; 6, 12 and 24 h for measurement of BMP-2 expression). NF-kappaB activation was identified as nuclear staining by immunohistochemistry. BMP-2 expression was observed through Western blot, immunohistochemistry and in situ hybridization. The results showed that: (1) Ang II stimulated the activation of NF-kappaB. Translocation of NF-kappaB p65 subunit from cytoplasm to nucleus appeared as early as 15 min, peaked at 30 min (P<0.01) and declined after 1 h. (2) TMP inhibited Ang II-induced NF-kappaB activation (P<0.01). (3) Ang II increased BMP-2 expression at 6 h but declined it significantly at 12 and 24 h (P<0.01). (4) BMP-2 expression was also kept at high level at 6 h in Ang II + TMP group but maintained at the normal level at 12 and 24 h. (5) There was no significant difference in NF-kappaB activation and BMP-2 expression between the control group and TMP group. These results indicate that TMP inhibits Ang II-induced VSMC proliferation through repression of NF-kappaB activation and BMP-2 reduction, and BMP-2 expression is independent of the NF-kappaB pathway. In conclusion, TMP has therapeutic potential for the treatment of atherosclerosis.
Angiotensin II ; antagonists & inhibitors ; Animals ; Atherosclerosis ; drug therapy ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; analysis ; antagonists & inhibitors ; Immunohistochemistry ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; NF-kappa B ; analysis ; antagonists & inhibitors ; Pyrazines ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; analysis ; antagonists & inhibitors

BACKGROUNDTo select the mutants of HPV type 16 E6 and E7 genes suitable for construction of vaccine for treatment of cervical cancer.

METHODSE6 and E7 genes were modified by site-directed mutagenesis. Several recombinant vaccina viruses were constructed by inserting the E6 or E7 mutants into the genome of vaccina virus Tiantan strain and employed to study their antigenicity.

RESULTSWestern blot assay showed that the E6 ?mutant? with substitution of Gly for Leu at amino acid site 50 and E7 mutant with substitution of Gly for Cys-24 and Glu-26 had no effect on their stability and antigenicity, but change of the Cys at position 91 of E7 dramatically reduced its stability and antigencity. Conclusion The results confirmed that the Zinc-finger structure at the E7 C-terminal? plays an important role in the integrity and stability of E7 protein.

Animals ; Antibodies, Viral ; biosynthesis ; Female ; Mice ; Mice, Inbred C57BL ; Mutagenesis, Insertional ; Oncogene Proteins, Viral ; genetics ; immunology ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Repressor Proteins ; Vaccinia virus ; immunology ; Zinc Fingers

Objective To construct the small interfering RNA (siRNA) targeting heparanase gene and its expressing vector,and to observe its interference effect on the expression of heparanase gene and inhibitory effect on the invasive potential of human malignant melanoma A375 cells.Methods Three siRNAs were designed.The recombinant plasmid pRNATU6.1/heparanase-siRNA was designed and constructed. A375 cells were cultured,and transfected with pRNATU6.1/heparanase-siRNA.The cells treated with lipo- fectamine or Opti-MEM served as the controls.Real-time fluorescence quantitative PCR and Western blot were performed to evaluate the expression of heparanase RNA and protein in these treated A375 cells.The in vitro invasive potential of treated A375 cells was assessed by Matrigel gel assay.Results The siRNA targeting heparanase gene was successfully cloned to the eukaryotic expressing vector pRNATU6.1.The expression levels of both heparanase RNA and protein decreased significantly in siRNA-transfected A375 cells than those in the control cells.The in vitro invasive potential of siRNA-transfected cells was also signifi- cantly inhibited as compared with that of the control cells (P

CEUS is a pure blood pool imaging depending on the non-linear effect and strong backscatter of gas-filled microbubbles in blood to obtain contrast-enhanced images.With the advantages of displaying tumor microvascular dynamically and intuitively,CEUS has been widely used for diagnosis and evaluating prognosis of breast cancers.CEUShas high sensitivity and specificity in detecting early breast cancers and identifying sentinel lymph nodes,with wide application prospects in assessing the efficacy of neoadjuvant therapy,predicting molecular typing and targeted therapy of breast cancers.The features of CEUS in breast cancers and the relationships with biomarkers,progresses in assessing neoadjuvant therapy and assisting sentinel lymph node biopsy were reviewed in the article.

Objective:To analyze the application value of narrative education model in clinical teaching of nurses in operating room.Methods:A total of 90 nursing students in the Department of Operating Room, Shanghai Tenth People's Hospital from January 2018 to January 2020 were selected as the research objects and randomly divided into observation group and control group, 45 in each group. The control group was taught by traditional mentoring method, and the observation group was taught by narrative education model. The two groups of nursing students practiced for 4 weeks. After the teaching, the scores of theoretical knowledge and operation were compared between the two groups. The scores of career identity scale and caring ability evaluation scale of the two groups were compared, the scores of each dimension of Chinese registered nurse core competence scale (CIRN) were evaluated, and the feedback of the two groups of nursing students on the teaching methods was analyzed. SPSS 22.0 was used to conduct t-test and chi-square test. Results:The scores of theoretical knowledge and operation in the observation group were (94.52±5.34) points and (88.67±3.98) points, respectively, which were significantly higher than those [(85.67±5.26), (82.24±4.16)] in the control group ( t=7.92, 7.49, all P<0.001). The scores of professional identity scale and caring ability evaluation scale in the observation group were (39.12±5.12) points and (193.14±11.24) points, respectively, which were significantly higher than those [(34.21±4.23), (180.23±11.35)] in the control group ( t=4.96, 5.42, all P<0.001). The scores of various dimensions of CIRN in the observation group were significantly higher than those in the control group ( t=4.72, 7.57, 4.13, 4.66, 6.19, 4.27, 5.48, all P<0.001). The feedback of nursing students in the observation group on teaching methods was significantly better than that in the control group ( χ2=4.29, 6.48, 12.26, 4.44, 7.60, 5.03, P<0.05). Conclusion:Narrative education model can significantly improve the internship effect of operating room interns, with clinical value.

Objective:To retrospectively analyze the experience of our center in the use of marginal donor heart, and to explore the principle of use and risk control of marginal donor heart.Methods:A total of 31 patients with end-stage heart disease underwent orthotopic heart transplantation in our center from January 2018 to December 2018, including 28 cases of pure heart transplantation, 2 cases of combined heart-lung transplantation, and 1 case of combined heart-kidney transplantation. 26 of the 31 cases were marginal donor hearts. These patients were all anastomosed by a double lumen method.Results:The rates of postoperative use of ECMO, IABP and acute rejection were zero in this study. The time of cardiopulmonary bypass in the marginal donor group was significantly longer compared with the conventional donor group( P<0.05), but there was no significant difference between the two groups in terms of hospitalization time, mechanical ventilation time, ICU stay time, abnormal rate of ECG, LVEF and blood biochemical indexes(all P>0.05). The postoperative follow-up rate was 100% in the two groups. One case of combined heart-lung transplantation in the marginal donor group died of multiple organ failure in the first month after surgery. During the postoperative follow-up period, the incidence of moderate to severe tricuspid regurgitation and the incidence of recurrent heart failure were zero in the two groups. There was no significant difference in the incidence of arrhythmia, LVEF, infection and blood biochemical parameters. Conclusion:The application of marginal donor heart has no significant effect on the short-term survival rate and recovery of patients after heart transplantation, but the long-term effect needs further follow-up.