Objective To evaluate the immunogenicity of a recombinant adenovirus containing the porin IB (PIB) gene and Neisseria surface protein A (NspA) gene of N.gonorrhoeae in BALB/c mice.Methods A recombinant adenovirus containing the PIB gene and NspA gene of N.gonorrhoeae (pAdEasy-1-PN) was constructed in previous studies.Thirty BALB/c mice were randomly and equally divided into 5 groups:low-,medium-and high-dose experiment group intramuscularly immunized with 104,106,and 108 50％ tissue culture infective dose (TCID50) of the recombinant adenovirus pAdEasy-1-PN,respectively,pAdEasy-1 control group immunized with 106 TCID50 of pAdEasy-1,blank control group immunized with sodium chloride physiological solution.Immunization was carried out twice at a 4-week interval.Serum samples were collected at 0,3 and 5 weeks after the first immunization,and spleens were removed at 5 weeks followed by the isolation of spleen lymphocytes.Enzyme linked immunosorbent assay (ELISA) was performed to determine the serum levels of PIB-specific and NspA-specific antibodies,methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferaton activity of spleen lymphocytes after stimulation by the recombinant PIB and NspA antigens.A slide agglutination test was used to estimate the anti-bacterial activity of murine serum.The complement-dependent bacteriolytic activity of murine serum was also evaluated.Statistical analysis was carried out by t test.Results Both humoral and cellular immune response specific to PIB and NspA were elicited by the recombinant adenovirus in mice.At 3 and 5 weeks after the immunization,significant differences were observed in the serum levels of PIB-specific (F =285.72,564.83,respectively,both P ＜ 0.01) and NspA-specific (F =521.57,542.61,respectively,both P ＜ 0.01)antibodies.Also,the proliferation index was statistically different among these groups in spleen lymphocytes stimulated with PIB and NspA (F =171.61,233.96,respectively,both P ＜ 0.01).The vaccination efficiency was positively correlated with the inoculation dose of recombinant pAdEasy-1-PN,and 108 TCID50 per dose proved to be the optimal dose for immunization.The sera from mice immunized with pAdEasy-1-PN could agglutinate N.gonorrhoeae and kill it in the presence of complement.Conclusions The recombinant adenovirus pAdEasy-1-PN containing PIB and NspA genes could induce specific humoral and cellular immune response in mice,and may be a potential vaccine against N.gonorrhoeae.

OBJECTIVETo observe the effect of Kuntai Capsule (KC) on the number of retrieved oocytes, the quality of high-quality oocytes and embryos in in vitro fertilization of poor ovarian response (POR) patients.

METHODSTotally 70 POR patients preparing for in vitro fertilization-embryo transfer (IVF-ET) were randomly assigned to the observation group and the control group, 35 cases in each group. KC was administered to patients in the observation group in the preparation cycle (i.e., three menstrual cycles before IVF-ET) and during the superovulation process. Those in the control group took placebo during this period. Before and after medication the improvement of Shen yin deficiency syndrome (SYDS) was observed in the two groups. The basal follicle-stimulating hormone (bFSH), luteinizing hormone (LH), estradiol (E2), anti-Mullerian hormone (AMH), the ratio of FSH to LH, and antral follicle count (AFC) were observed. Besides, the E2 level of a single ovum on the day of HCG injection, the number of retrieved oocytes, the high-quality oocyte rate, and the high-quality embryos were observed.

RESULTSCompared with the control group, the SYDS, decreased bFSH and LH levels, increased ACF numbers, the E2 level of a single ovum on the day of HCG injection, the number of retrieved oocytes, high-quality oocytes, and high-quality embryos were superior in the observation group (P < 0.05). There was no statistical difference in the decreased FSH/LH level (P > 0.05). E2 and AMH increased after medication of KC in the observation group, while they decreased after administration of placebos in the control group. There was statistical difference in the post-pre treatment difference of E2 and AMH between the two groups (P < 0.05).

CONCLUSIONKC could increase the number of retrieved oocytes, and elevate the quality of occytes and embryos in the IVF-ET.

Adult ; Drugs, Chinese Herbal ; pharmacology ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Oocyte Retrieval ; Oocytes ; drug effects ; Pregnancy

ObjectiveTo evaluate the attachment type,the level of alexitymia and the defense style of depression patients and their relativity.Methods 32 depression patients were assessed with the adult attachment scale (AAS-1996 Revised Edition),toronto alexithymia scale (TAS) and defense style questionnaire( DSQ),and 80 healthy people were involved as controls.Results The depression patients showed significantly different attachment type from the control group( x2 =47.6,P ＜ 0.01 ).Compare with the control group,the depression patients showed higher scores in immature defense style( (5.47 ± 1.13 ) vs (3.79 ± 1.09 ) ) and neurotic defense style( (4.93 ±0.56) vs (4.20 ± 1.00),(P＜0.01) ).Compared with controls,the depression patients showed higher scores in TAS total score( (76.88 ± 8.94) vs (65.70 ± 7.98 ) ) factor Ⅰ ( ( 3.34 ± 0.77 ) vs ( 2.41 ±0.78)) and factor Ⅱ ((3.64 ±0.51) vs (2.65 ±0.55),P＜0.01),and lower scores in factor Ⅲ((2.51 ±0.78) vs (3.14 ± 1.03 ),P ＜ 0.01 ).The depression patients' adult attachment was significantly correlative with their scores of alexithymia.The depression patients' adult attachment was also significantly correlative with their scores of defense style.ConclusionThe attachment of depression patients may lead to depression via their defense style.Depression leads to alexitymia.Depression patients with different features of attachment show different level of alexitymia.

Objective To investigate the clinical significance between congestive heart failure (CHF) and serum thyroxine. Methods 50 patients with CHF (7 cases of grade Ⅱ heart function,grade Ⅲ 19 cases,grade Ⅳ 24)and 30 normal persons wre selected. The serum triiodothyronine( T3 ), four-triiodothyronine( T4 ), free triiodothyronine (FT3), free triiodothyronine (FT4), thyroid stimulating hormone TSH were determined by radioimmunoassay. Results The T3, FT3 contens ( 1.95 ± 0.55) nmol/L, (2.20 ± 0.39 ) pmol/L in CHF group were lower than the control group (2.4 ± 0.50) nmol/L, ( 5.89 ± 0. 45 ) pmol/L ( t = 2. 415,2. 325, P < 0.05 ); The T3, FT3 contens ( 2.19 ± 0. 53 )nmol/L,(3.69 ± 0.57)pmol/L after treatment were significantly higher than that before treatment (t =2. 375,2. 405, P < 0.05 ); The T3, FT3 contens in cardiac function with stage Ⅳ ( 1.76 ± 0.7) nmol/L, ( 1.99 ± 0. 45 ) pmol/L were significantly lower than the control group( t = 2.245,2. 375, P < 0.05 ); Serum TT3, TT4 and cardiac function was negatively correlated ( r = -0.933, -0.383, all P < 0.05 ) , serum FT3, FT4,TSH and cardiac function were not correlated. Conclusion The changes of serum thyroid level could be used as an index of the severity of heart failure.

This paper reported 13 cases of delayed traumatic chest wall abscess from January 2012 to January 2015.All the patients were associated with type 2 diabetes.After local puncture for confirmative diagnosis, a chest wall abscess dissection was carried out as soon as possible.At each site of upper and lower pole, an indwelling drainage tube was placed for irrigation and negative pressure suction.Sensitive antibiotics were selected based on susceptibility test results.The drainage tubes were removed 7-14 days after surgery.There were 8 cases of primary healing of incision and 5 cases of secondary healing of incision.All the patients were cured.Follow-ups for 6-36 months (average, 17 months) showed no recurrence.

Animals ; Hindlimb ; blood supply ; Ischemic Preconditioning ; Lung ; metabolism ; Male ; P-Selectin ; metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism

AIM: To express the DT389-hbFGF (389 amino acid residues of the N-terminus of diphtheria toxin (human basic fibroblast growth factor) fusion protein for potential targeting therapy towards posterior capsule opacification (PCO) after cataract surgery.METHODS: The DNA of inactivated diphtheria bacillus and RNA of 12-week fetal brain cortex were extracted, respectively. The fragments of truncated diphtheria toxin (containing 389 amino acids of N-terminus, DT389) )and full-length human basic fibroblast growth factor(hbFGF) sequence (encoding 18kDa protein) were amplified by PCR. The two fragments were inserted into pGEX-4T-1 prokaryotic expression vector to obtain pGEX-DT389-hbFGF prokaryotic expression plasmid. After sequence analysis, the expressing plasmid was transformed into Escherichia Coli BL21 strain and expression was induced under IPTG. The expressed fusion protein was purified and identified.RESULTS: The gene fragments encoding DT389 and hbFGF were amplified and their gene sequences were confirmed. Hybrid gene expression plasmid pGEX-DT389 (hbFGF was constructed. The fusion protein DT389-hbFGF was expressed and purified.CONCLUSIONS: The successful cloning and expression of DT389-hbFGF immunotoxin provides a foundation for targeting therapy towards posterior capsule opacification.

Multiplicity of signals and diversity of signaling pathways exist during the establishment of mycorrhizal associations together with the regulation of symbiosis-specific genes expression.This mechanism of signal recognition and transduction related with development process of the symbiont was reviewed at the molecular level.

Objective:To evaluate the effect of post-conditioning in brain injury induced by myocardial I/R on inflammatory factor and GFAP.Methods:Male Sprague-Dawley rats were randomly allocated into 3 groups (n=8):group Sham,group IR,group IPost.Myocardial IR was induced by occlusion of the anterior descending branch of the left coronary artery for 30 min.group IPost received 3 cycles of 10 s reperfusion followed by 10 s ischemia at the end of myocardial ischemia.The rats were sacrificed at 120 rain of reperfusion and the brains were removed for microscopic examination,inflammatory factors and GFAP.Results:Compared with group Sham,IL-6,IL-8 were significantly increased,IL-10 was down-regulated in group IR(P＜0.01).Post-conditioning can decrease IL-6,IL-8 and up-regulated IL-10(P＜0.01).When compared with group Sham,the expression of GFAP was higher in group IR(P＜0.05),however,the GFAP in group IPost is the most among these three groups(P＜0.01).Conclusion:Post-conditioning could protect brain by decreasing inflammatory factors,increasing GFAP,which both from brain injury induced by myocardial ischemia reperfusion.

BACKGROUND:Neurotrophin-3 is found in the repair of spinal cord injury in the role of the strongest neurotrophic factor.It can effectively promote axonal regeneration through the glial scar tissue in repairing spinal cord injury.OBJECTIVE:To construct recombinant lentiviral vectors for gene delivery of homo sapiens neurotrophin-3(hNT3),and to investigate the expression of hNT3 gene in Schwann cells after transfection.DESIGN,TIME AND SETTING:Observational experiment was performed from June 2007 to March 2008 at the Central Laboratory of Changhai Hospital.MATERIALS:Bilateral sciatic nerves were harvested from 3-day-old Sprague Dawley rats for culture and identification of Schwann cells.Three-plasmid lentivirus systems:pGC-E1-EGFP,pHelper 1.0 and pHelper 2.0 were gained from Shanghai Chemical Technology Co.,Ltd.METHODS:pGC-E1-hNT3-EGFP plasmid was constructed by double restriction enzyme digestion and ligation,and then the plasmid was transformed into E.coli DH5?.Purified pGC-E1-hNT3-EGFP plasmids from the positive clones was confirmed by PCR and sequencing.293T cells were cotransfected with lentiviral vector pGC-E1-hNT3-EGFP,pHelper 1.0 and pHelper 2.0 by Lipofectamine 2000 to produce lentivirus.2 mL recombinant virus complete culture solution was added according to multiplicity of infection=1,4,8,10,12.The titer of virus was tested according to the expression level of enhanced green fluorescent protein.The control groups were Schwann cells and Schwann cells transfected by no-loaded lentivirus.MAIN OUTCOME MEASURES:The lentiviruses were transduced to Schwann cells,and the transfection efficiency was examined by flow cytometry,the overexpression of hNT3 was determined by Real-time PCR and Western blotting.RESULTS:The exogenous gene sequence of the recombinant hNT3 was completely in accordance with that of its open reading frame in GeneBank.The titer of concentrated virus was 5?107 TU/L.After recombinant LV-hNT3 infection,Schwann cells gave off strikingly bright green fluorescence,and the transfection efficiency amounted to 85%(multiplicity of infection=10).Real-time RCR test showed that the hNT3 mRNA were highly expressed in hNT3-Schwann cells,while did not express in the control group.Western blotting showed the hNT3 expression in Schwann cells.CONCLUSION:Construction of hNT3 gene lentiviral vector can transfect Schwann cells leading to an efficient overexpression of hNT3.