OBJECTIVE To study the expression situation of esp gene and the relativity between esp gene and resistance of Enterococcus faecalis derived-infection.METHODS The E.faecalis esp gene was amplified by PCR,then it was cloned and sequenced;the minimum inhibitory concentration(MIC) of five antimicrobicals against E.faecalis was determined by agar dilution method.RESULTS The positive rate of esp gene in 61 strains of E.faecalis was 42.6%;the positive rate of esp gene in the resistant strains against ampicillin,ciprofloxacin,high-level gentamicin,and erythromycin was 33.3%,54.8%,70.6%,and 51.0%,respectively,and the positive rate of esp gene in the susceptible strains against above 4 antibacterials was 43.6%,30.0%,7.4%,and 8.3%,respectively.CONCLUSIONS Incidence of esp gene does not correlate with the resistance of E.faecalis against ampicillin,but clearly correlate with the resistance of E.faecalis against ciprofloxacin,high-level gentamicin,and erythromycin;esp could become a marker of high-level gentamicin against E.faecalis derived-infection.

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

Objective To find out the iodine nutritional status of Longyan downtown residents,evaluate the effectiveness of control measures and provide a scientific basis for developing control strategies.Methods Infants aged 0 to 2 year-old,children aged 8 to 10,adults aged 18 to 45 and pregnant and lactating women were selected as survey subjects.Children goiter was detected with B ultrasound.Residents per capita daily salt intake was investigated by weighing method.Three urinary samples and a milk sample of lactating women were randomly collected.Urinary iodine and milk iodine content were determined by arsenic cerium catalytic spectrophotometric assay.Blood samples were collected and thyroid function (including serum TT3,FT3,TT4 and FT4) and thyroid stimulating hormone(TSH) were measured with direct chemiluminescence immunoassay,and thyroglobulin antibody(TgAb),thyromicrosome antibody(TMAb),and thyroglobulin (Tg) were measured with radioimmunoassay (RIA) in serum.ResultsThe goiter rate of children aged 8 to 10 was 1.8％ (2/110),and median thyroid volume was 2.75 ml.Household iodized salt coverage rate was 100.00％(318/318),and qualified iodized salt was 94.03％ (299/318).The daily per capita salt intake was (6.13 ± 3.56)g.The average medians of urinary iodine of the infants,children,adults,pregnant and lactating women were 181.8,315.2,196.6,158.7,136.4 μg/L,respectively.The median of milk iodine of lactating women was 155.6 μg/L.The proportions of serum TT3,FT3,TT4,FT4 and TSH which higher than normal were 3.6％ (11/308),0.6％ (2/309),23％ (7/309),1.0％ (3/313) and 1.3％ (4/312),respectively.While the proportions of serum TT3,FT3,TT4,FT4 and TSH that lower than normal were 2.3％ (7/308),11.7％(36/309),2.3％(7/309),12.8％(40/313),and 1.6％(5/312),respectively,of which 16 cases of both TgAb and TMAb were higher than normal.ConclusionsExisting salt iodine level is appropriate for 0 to 2 year-old infants and young children,18 to 45 year-old adults,pregnant and lactating women in downtown Longyan city.The iodine intake of children aged 8 to 10 is excessive.Thyroid function monitoring is recommended to be included in the routine monitoring.

Objective To summarize the clinical experience and efficacy of surgical treatment for Stanford type A aortic dissection leading to acute lower limb ischemia.Methods From January 2014 to January 2018,12 patients with severe lower limb ischemia caused by acute type A aortic dissection were treated with Suns surgery.Among them,11 patients were treated with restoration of lower limb blood supply preferentially,including 10 cases of femoral artery bypass and 1 case of abdominal aorta-iliac artery stent graft implantation.Another case was treated with ascending aorta-femoral artery bypass after Sung surgery.Results 3 cases died of ischemia and necrosis of the lower extremities.Two of them died of multiple organ failure due to amputation and one died of low cardiac output due to refractory acidosis.Acute renal failure performed bedside CRRT in 5 patients and ECMO in 1 patient.The remaining 9 patients were discharged from the hospital and the symptoms of lower limb ischemia disappeared.After an average follow-up of 23 months,the re-examination of the aorta CTA showed that the bypass artery was unobstructed and the distal femoral artery was well developed.One patient infecting vascular prosthesis was cured by taking out the unit.Conclusion For acute lower limb ischemia caused by type A aortic dissection,blood flow of lower extremities should be restored as soon as possible to reduce mortality and complications.Femoral artery bypass and abdominal aorta-iliac arterial repair are simple and effective in reconstructing lower limb blood supply.

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.

Background: Feline mammary carcinoma is the third most common cancer that affects female cats. Objectives: The purpose of this study was to screen differential serum proteins in feline and clarify the relationship between them and the occurrence of feline mammary carcinoma. Methods: Chinese pastoral cats were used as experimental animals. Six serum samples from cats with mammary carcinoma (group T) and six serum samples from healthy cats (group C) were selected. Differential protein analysis was performed using a Label-free technique, while parallel reaction monitoring (PRM) was performed to verify the screened differential proteins. Results: A total of 82 differential proteins were detected between group T and group C, of which 55 proteins were down regulated and 27 proteins were up regulated. Apolipoprotein A-I, Apolipoprotein A-II (ApoA-II), Apolipoprotein B (ApoB), Apolipoprotein C-III (ApoC-III), coagulation factor V, coagulation factor X, C1q, albumen (ALB) were all associated with the occurrence of feline mammary carcinoma. Differential proteins were involved in a total of 40 signaling pathways, among which the metabolic pathways associated with feline mammary carcinoma were the complement and coagulation cascade and cholesterol metabolism. According to the Label-free results, ApoB, ApoC-III, ApoA-II, FN1, an uncharacterized protein, and ALB were selected for PRM target verification. The results were consistent with the trend of the label-free. Conclusions This experimen is the first to confirm ApoA-II and ApoB maybe new feline mammary carcinoma biomarkers and to analyze their mechanisms in the development of such carcinoma in feline.

OBJECTIVETo evaluate the effects of antiarrhythmic peptide (AAP10) on ventricular arrhythmias in rabbits with healed myocardial infarction (OMI).

METHODSThirty rabbits were randomly divided into three groups (n = 10 each): Sham group, left thoracotomy was performed without coronary ligation; OMI group and OMI + AAP10 group, the circumflex coronaries were ligated. Three months post operation, the electrophysiological and antiarrhythmic effects of AAP10 were assessed in the arterially perfused rabbit left ventricular wedge preparation. Sham and OMI group were perfused with Tyrode's solution and OMI + AAP10 group was perfused with Tyrode's solution + AAP10 (80 nmol/L). Transmembrane action potentials were recorded simultaneously from endocardium and epicardium together with a transmural ECG by use of 2 separate intracellular floating microelectrodes. The stimulus-response-interval (SRI) of the epicardium and the incidence of ventricular tachycardia (VT) were observed. Whole heart and left ventricular weights, the left ventricular thickness at infarct border zone were measured.

RESULTSWhole heart and left ventricular weights as well as the left ventricular thickness at the infarct border zone significantly increased post infarction. VT was induced in 8 out of 10 rabbits in OMI group and in 2 out of 10 rabbits in OMI + AAP10 group (P < 0.05). SRI was also significantly shortened in OMI + AAP10 group compared to OMI group [SRI-1: (20.59 +/- 0.79) ms vs. (28.71 +/- 0.55) ms; SRI-2: (30.42 +/- 0.74) ms vs. (38.67 +/- 0.49) ms, all P < 0.01]. However, the action potential morphology and duration were similar between OMI and OMI + AAP10 groups.

CONCLUSIONThe antiarrhythmic peptide (AAP10) can increase gap junctional intercellular conductance without affecting the action potential morphology and duration and decrease the incidence of inducible ventricular tachycardia.

Animals ; Arrhythmias, Cardiac ; etiology ; prevention & control ; Male ; Myocardial Infarction ; complications ; physiopathology ; Oligopeptides ; pharmacology ; Rabbits ; Random Allocation

OBJECTIVETo investigate the anti-tumor immunity of the non-replicating recombinant vaccinia virus expressing HPV16 E6 and E7 proteins.

METHODSC57BL/6 mice were immunized by non-replicating recombinant vaccinia virus (NTVJmE6E7), and then specific CTLs were determined. Immune protection effects were evaluated by challenges of different doses of TC-1 tumor cells. Immunotherapeutic effects in form of recurrence were evaluated on the tumor-removed mice.

RESULTSMice immunized by NTVJmE6E7 could generate TC-1 cell specific cytotoxic T lymphocyte (CTL). Mice boosted with NTVJmE6E7 could tolerate the challenge of 1 x 10(4) TC-1 cells. NTVJmE6E7 could effectively prevent the tumor recurrence in the tumor-removed mice.

CONCLUSIONNTVJmE6E7 can be taken as a candidate of therapeutic vaccine for HPV-associated tumors and their precursor lesions.

Animals ; Cancer Vaccines ; Cells, Cultured ; Female ; Genetic Vectors ; Immunotherapy ; Lymphocyte Activation ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Neoplasms, Experimental ; immunology ; therapy ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Recombination, Genetic ; Repressor Proteins ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccinia virus ; genetics ; Viral Vaccines

OBJECTIVETo generate a human papillomavirus (HPV16) prophylactic and therapeutic vaccine candidate for cervical cancer.

METHODSHPV16 major capsid protein L1 gene/minor capsid protein L2 gene and HPV16 early E6/E7 genes were inserted into a vaccinia virus expression vector. A strain of non-recombinant vaccinia virus containing the sequences was obtained through a homologous recombination and identified.

RESULTSDNA hybridization confirmed that the HPV16L1/L2/E6/E7 genes were integrated into vaccinia virus DNA. Western Blot result showed that full-length L1/L2/E6/E7 proteins were co-expressed in CEF cells infected with the recombinant virus.

CONCLUSIONNTVJE6E7CKL1L2 could be taken as a candidate of prophylactic and therapeutic vaccine for HPV-associated tumors and their precancerous transformations.

Animals ; Blotting, Western ; Capsid Proteins ; genetics ; metabolism ; Cells, Cultured ; Chick Embryo ; Cloning, Molecular ; Female ; Gene Expression ; Genetic Vectors ; genetics ; Humans ; Oncogene Proteins, Viral ; genetics ; metabolism ; Papillomaviridae ; genetics ; immunology ; Papillomavirus E7 Proteins ; Papillomavirus Infections ; immunology ; prevention & control ; virology ; Papillomavirus Vaccines ; genetics ; immunology ; therapeutic use ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Transfection ; Tumor Virus Infections ; immunology ; prevention & control ; virology ; Uterine Cervical Neoplasms ; immunology ; prevention & control ; virology ; Vaccinia virus ; genetics ; Virus Replication

Objective To study and research the transcription pattern of UL131A-128 mRNA in human cytomegalovirus (HCMV) clinical low passage strains.Methods The UL131 A-128 mRNAs of from different clinical strains and kinetic periods were amplified using 3 ' RACE and analyzed by sequencing.Meanwhile,clones containing UL131A-128 transcripts in a HCMV cDNA library of clinical strain were selected and sequenced.Results It is successful to obtain the transcription pattern of UL131 A,UL130 and UL128 gene in HCMV clinical low passage strains,the UL131A gene consisted of two exons and the coding region of UL130 gene was not interrupted by any intron in the region as reported before.However,the transcript of UL128 gene showed two patterns,one pattern consisted of the three exons,the length is 519bp,and the other one contained the three exons and the sequence of the first intron further,the length is 642bp.The quantities of UL128 transcript containing the sequence of the first intron were higher than that of transcript only containing the three exons in the studied clinical strains at all kinetic classes.It was demonstrated that the UL131A-128 mRNA were expressed with immediately early,early and late kinetics.The result of 3'RACE and HCMV cDNA library of clinical strain is conformity.Conclusions Our results demonstrated that the UL1 31 A,UL130 and UL128 genes were transcribed with 3'-coterminal,although the initiation points of their mRNA may be different.The variation of the transcripts found in our study indicated complex nature of transcription of UL131A-128 genes in HCMV clinical strains.