Tuberculosis (TB) remains an enormous health burden worldwide.To date,Mycobacterium bovis Bacillus Calmette Guerin (BCG) is the unique anti-TB vaccine available for humans,which provides an important but limited protection from the Mycobacterium tuberculosis (Mtb) infection.It is therefore an urgent need to develop better vaccines and vaccination strategies to prevent the spread of Mtb infection.Heterologous prime-boost vaccination strategies using both BCG and novel anti-TB vaccines have been demonstrated to induce robust immune responses than BCG alone.We have previously demonstrated that a recombinant adenoviral vector Ad5-CEAB co-expressing CFP10,ESAT6,Ag85A and Ag85B of Mtb was able to induce robust antigen-specific immune responses in mice.In the present study,we examined immunological effects of Ad5-CEAB in the mice primed with BCG and boosted with a single dose of the virus via an intranasal route.Results demonstrated that this vaccination strategy could effectively induce strong antigen-specific mucosal and humoral immune responses.These immune responses were characterized with an increased productions of cytokines (IL-12 and IFN-γ),increased concentration of secretary IgA (sIgA) in bronchoalveolar lavage fluid (BALF) and serum IgG in mice in comparison with mice in BCG group.These data suggested that the regimen of BCG prime-single dose of Ad5-CEAB boosted strategy was novel for inducing antigen-specific immune responses in response to Mtb antigens in vivo,which warrants for further development of adenoviral-based vaccine against Mtb infection.

Objective To gain more pathophysiolgic knowledge about acute obstructive hydrocephalus and to explore its rapid and effective treatment by establishing canine acute obstructive hydrocephalus model.Methods Acute obstructive hydrocephalus model was established by injecting cyan-acrylic gel glue into the fourth ventricle via posterior fosse craniotomy in 9 male adult mongrel dogs.At the same time,lateral ventricle catheterization were performed and were fixed on the scalp to connect reservoir bag so that the changes of intracranial pressure (ICP) could be measured dynamically,and the changes of neurological function were observed.Results Acute obstructive hydrocephalus model was successfully established in 6 of the total 9dogs.ICP was (48.2 ± 6.1 ) cm H2 O at 48 hours after the injection and was (56.4 ± 5.7 ) cm H2 O at 72 hours after the injection,it increased 392％ and 459 ％ respectively.And the ICP after injection was significantly different(P ＜ 0.01 )compared with that before injection (12.3 ± 3.1 )cm H2O.Conclusion The establishment of acute obstructive hydrocephalus model has high success rate,and is easy to reduplicate; ICP could be measured dynamically and also could be reduced by releasing CSF;Thus,ventricular drainage is the most rapid and effective treatment for acute obstructive hydrocephalus.

CFP10, ESAT6, Antigen 85A (Ag85A) and antigen 85B (Ag85B) are the key immunodominant antigens of Mycobacterium tuberculosis. In order to construct a eukaryotic vector able to co-express the four genes in one vector, we amplified the target gene fragments encoding the CFP10, ESAT6, Ag85A and Ag85B antigens and inserted them into the multicloning site of the shuttle plasmid vector pcDNA3.1 (+), of which the CFP10 and ESAT6 encoding genes were in frame fused with a linker encoding (Gly4Ser)3 residue, before the fused gene was inserted downstream of CMV promoter with a bovine growth hormone poly A(BGH pA) sequence at the 3'-end; Ag85A and Ag85B encoding genes were fused with a separation of internal ribosome entry site (IRES) sequence before the fused gene cassette was inserted downstream of RSV promoter with a BGH pA sequence at the 3'-end. The final plasmid containing all four genes was confirmed by sequence analysis and designated as pcDNA-CFP10-ESAT6-Ag85A-Ag85B (pcDNA-CEAB). In order to verify the ability of this construct to express target proteins, we then transfected the recombinant plasmid into Human embryonic kidney (HEK) 293T cells and harvested the cell lysates, and the cell lysates were then separated by SDS-PAGE and subjected to Western blot analysis 48 h after transfection. All four of the target proteins were detected in the cell lysates against the respective specific antibodies, suggesting that we have successfully constructed a eukaryotic vector co-expressing the four immunodominant antigens of Mycobacterium tuberculosis, which lay a foundation for the further study of the immunogenicity and protective activity of the four antigens.
Acyltransferases ; biosynthesis ; Antigens, Bacterial ; biosynthesis ; Bacterial Proteins ; biosynthesis ; Genetic Vectors ; HEK293 Cells ; Humans ; Immunodominant Epitopes ; Mycobacterium tuberculosis ; Plasmids

BACKGROUND:Pulsed laser deposition synthesis technology has been used to prepare new nano-hydroxyapatite thin film coating by colagen deposition on artificial mechanical heart valve. OBJECTIVE: To investigate the toxicity of new nano-hydroxyapatite thin film on human umbilical vein endothelial cels. METHODS: Human umbilical vein endothelial cels were cultured with nano-hydroxyapatite film room-temperature leaching solution, nano-hydroxyapatite film high-temperature leaching solution, high-density polyethylene and phenol solution. Within 72 hours, cel growth was observed under the inverted phase contrast microscope. At 7 days after culture, cel proliferation and toxicity grading were detected using Cel Counting Kit-8. RESULTS AND CONCLUSION:At 24 hours after culture, cels grew wel, showed fusiform shape, and had strongrefraction in the nano-hydroxyapatite film room-temperature leaching solution, nano-hydroxyapatite film high-temperature leaching solution, high-density polyethylene groups, and no significant differences in cel morphology and number were detected among above groups. Cels in the phenol solution group were suspended, round, pyknotic and dead. At 48 hours, except phenol solution group, cel number increased significantly, and cel grew densely in other three groups. At 72 hours, cels grew strongly, and the gap became smal obviously. Within 7 days after culture, cel proliferation activity was not significant in the nano-hydroxyapatite film room-temperature leaching solution, nano-hydroxyapatite film high-temperature leaching solution, and high-density polyethylene groups, which was significantly higher than in the phenol solution group (P < 0.05). The toxicity of nano-hydroxyapatite film graded 0 to 1. These results suggested that nano-hydroxyapatite artificial mechanical heart valve has good histocompatibility, but no toxicity.

BACKGROUND:A new type of nano-hydroxyapatite artificial mechanical heart valve has been developed using pulsed laser deposition technology at the Department of Materials, Hefei University and Anhui Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, China. OBJECTIVE:To investigate the compatibility of nano-hydroxyapatite artificial mechanical heart valve with human umbilical vein endothelial cels. METHODS:Human umbilical vein endothelial cels were in vitroisolated, cultured and passaged to the 2-4 generations, and then the cel suspension was inoculated onto the nano-hydroxyapatite artificial mechanical heart valve. After 3, 7, 12 days of culture, the cel growth on the artificial mechanical heart valve was observed under scanning electron microscope. In addition, the human umbilical vein endothelial cels were respectively cultured in room-temperature and high-temperature extract liquids of nano-hydroxyapatite artificial mechanical heart valve, high-density polyethylene and phenol solution extracts for 72 hours, and then, the proliferation of cels was detected by MTT method. RESULTS AND CONCLUSION:Under the scanning electron microscope, the human umbilical vein endothelial cels were fusiform- or polygon-shaped with protuberances adhered to the artificial mechanical heart value at 3 days of culture; the cels were stretched thoroughly and fused at 7 days of culture; and the cels were confluent to pieces that tightly overlaid the heart valve surface and the extracelular matrix was formed localy at 21 days of culture. Results from MTT test displayed that the nano-hydroxyapatite artificial mechanical heart valve had no cytotoxicity to the human umbilical vein endothelial cels, indicating a good cytocompatibility.

BACKGROUND:In early experiments, we prepared hydroxyapatite-coated mechanical heart valve embedded with col agen using impulse laser sediment method.
OBJECTIVE:To further analyze the histocompatibility and toxicity of hydroxyapatite-coated mechanical heart valve embedded with col agen.
METHODS:After passage, canine vascular endothelial cellsuspension was inoculated onto the hydroxyapatite-coated mechanical heart valve embedded with col agen. One group was inoculated in 5%CO2, 37 ℃ incubator for 3 weeks static culture, and the other group was inoculated in 5%CO 2 , 37 ℃ incubator for 3 weeks spinner culture. Scanning electron microscope was used to observe cellattachment on the material. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to the proliferative capacity of vascular endothelial cells cultured with the hydroxyapatite-coated mechanical heart valve embedded with col agen.
RESULTS AND CONCLUSION:During the spinner culture, adherent cells were found on the surface of mechanical heart valve, and the cells distributed evenly and confluent at 21 days to cover the surface of the material. The number of adherent cells in the spinner culture was higher than that in the static culture. The cells during the static culture were aggregated and distributed irregularly. The mechanical heart valve exhibited no effects on the proliferation of canine vascular endothelial cells which grew wel . These findings indicate that the hydroxyapatite-coated mechanical heart valve embedded with col agen exert no effect on proliferation of vascular endothelial cells, has no toxicity and has good biocompatibility.

Objective To approach the normal manifestations of the hypopharynx of the adults at the level of cricoid cartilage as seen on CT images, and to evaluate their clinical value. Methods Eighty-four CT images of the normal hypopharynx were reviewed. The hypopharynx was divided into three regions:(1) the piriform sinus, (2) the retropharynx area, and (3) the postcricoid region. The postcricoid region of hypopharynx was subdivided into three levels as follows : (1) the upper margin slice of the cricoid cartilage at the cricoarytenoid joint level, (2) the middle portion slice of the cricoid cartilage, and (3)the inferior margin slice of the cricoid cartilage. The anteroposterior and transverse diameters, and anterior and posterior wall thicknesses in the postcricoid region were measured. Depiction of the layers of the musculature and adjacent fat planes was evaluated. Statistical comparisons of measured results were made by using ttest and x2 test. Results The posterior wall tended to be (0. 9±0. 4) mm thicker than the anterior wall. The average transverse extension of the postcricoid musculature was (4. 5±0. 3)mm shorter in female than that in male at CT images. There were statistically significant differences related to sex (at the upper margin level of the cricoid cartilage : the transverse extension was (38. 6±3. 3)mm in male, (34. d±2. 5) mm in female, t = 6. 26,P < 0. 05 ; at the middle portion level of the crieoid cartilage: (33.6±3. 6) mm in male,(28.9±2.8) mm in female t =6.36, P <0.01;at the inferior margin level of the cricoid cartilage:(28.6 ±3. 1) nun in male, (24. 0 ±2. 1) mm in female, t = 7.52, P <0. 01). The transverse diameter tended to taper (10. 1±2. 4) mm from the upper cricoid slice level to the lower cricoid slice levelDemonstration of the intramural fat planes of the postcricoid region decreased from the upper [ 81.0%(68/84)] to the lower region [23.8% (20/84)] of the cricoid cartilage. In fat planes around the postcricoid region at all levels, the posterior fat plane was seen least frequently, and the left-sided fat plane was seen most frequently. There was a statistically significant difference at every level(at the upper margin level of the cricoid cartilage, the visibilities of the left-sided, the right-sided, and the posterior fat plane around the postcricoid region were 77.4% (65/84), 72. 6% (61/84), and 28. 6% (24/84) ,x<'2> =24. 64,P <0.01 ; at the middle portion level of the cricoid cartilage, the visibilities were 89.3% (75/84), 75.0%(63/84), and 34.5% (29/84) ,x<'2> =24. 76, P <0. 01 ; at the inferior margin level of the cricoid cartilage:the visibilities were 95.2% (80/84), 88. 1% (74/84), and 52. 4% (44/84), x<'2> = 13.59, P < 0. 01.Conclusion Knowledge of the normal appearances and variations of the hypopharynx at the level of cricoid cartilage is essential in detecting abnormalities in this area.

Objective To summarize and analyze clinical diagnosis and surgical treatment methods of 11 cases with congenital coronary arterial fistula (CCAF). Methods The clinical data of 11 patients who were definited by ultrasonic cardiogram, CT angiography (CTA) and coronary angiography were analyzed retrospectively. Four cases were simple CCAF, 7 cases coexisted with other heart abnormalities. Six cases were given surgical closure of fistula without cardiopulmonary bypass. One of the cases adopted coronary artery under the tangent cotton-padded mattress suture, and 4 cases fistula arterial were ligatured directly. Six cases were given surgical closure of fistula under cardiopulmonary bypass. The right coronary arterial was opened in three of the cases with right coronary artery aneurysm to close fistula. The chambers of heart in the others were opened to close fistula. Results All patients received surgical treatment successfully, and no death happened during the operation. The ultrasonic cardiography showed that all patients recovered well. Follow-up was conducted on 10 patients with the time period ranging from 3 months to 5 years. There was no death and no complication. Conclusions Combined application of ultrasonic cardiogram, coronary angiography and CTA increases the accuracy rate of diagnosis greatly and offers visual bases to formulate operation plan. Surgical operation is quite effective for congenital coronary arterial fistula after definite diagnosis. Operator should try to reserve the expanded coronary arterial, strengthen the anticoagulant after opeation to prevent thrombosis.

Objective To summarize the susceptible factors and the experience of the surgical treatment of ruptured aneurysm of the sinus of Valsalva combined with infective endecarditis.Methods From January 2000 to March 2008,30 cases with raptured aneurysm of the sinus of Valsalva were admitted, clinical data of 10 cases combined with infective endocarditis were retrospectively reviewed.Nine cases underwent the surgical treatment,including 6 cases accepted aortic valve replacement (AVR).Results Nine eases underwent the surgical treatment followed up for 2 months to 8 years,cardiac function of the survivors recovered to Class Ⅰ-Ⅱ in NYHA,7 cases were Ⅰ grade,2 cases were Ⅱ grade.One case was dead for MSOF not underwent the surgical treatment.Conclusions Associated with ventricular septal defect is one of the susceptible factors.Following diagnosis of ruptured aneurysm of sinus of Valsalva combined with infective endocarditis,sufficiently and validly antibiotic and surgical treatment should be pedormed as quickly as possible,and AVR to the moderate and severe aortic insufficiency.

OBJECTIVETo study the effect of retinoid kappa receptor alpha (RXRalpha) transfection plus treatment with the RXRalpha ligand, 9-cis-RA, on the proliferation and phenotype of platelet-derived growth factor (PDGF) activated hepatic stellate cells (HSCs).

METHODSPDGF activated rat hepatic stellate cells were transfected with eukaryotic expression vector pcDNA3.1- human RXRalpha, and confirmed by Western blot. Proliferation of transfected HSC was assayed by bromodeoxyuridine (BrdU) incorporation as well as MTT, and the phenotype (alpha-smooth muscle actin, desmin) was observed by immunocytochemistry with image analysis.

RESULTSTransfection of the RXRalpha gene and treatment with ligand 9-cis-RA of PDGF-activated HSCs extended the increased expression of RXRalpha protein for at least 168 hours. Cell proliferation and expressions of alpha- smooth muscle actin (alpha-SMA) and desmin were blocked, compared with groups of sham-transfected, PDGF-activated, no transfection, no ligand treatment, and irrelevant ligand treated HSCs.

CONCLUSIONTransfection with the RXRalpha gene followed by 9-cis-RA ligand treatment will inhibit the proliferation and reverse the phenotype of activated HSC.

Animals ; Cell Division ; Cells, Cultured ; Liver ; cytology ; Liver Cirrhosis ; etiology ; Male ; Phenotype ; Platelet-Derived Growth Factor ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Retinoic Acid ; genetics ; physiology ; Retinoid X Receptors ; Transcription Factors ; genetics ; physiology ; Transfection