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Objective:To construct and purify four respiratory syncytial virus (RSV) PreF proteins through gene sequence design and optimization and evaluate their immunogenicity.Methods:Coronin-1A and T4 trimer protein gene sequences were optimized with Human and CHO codons, and then added to RSV F protein sequence. The above plasmids were transfected into Expi293F cells for protein expression. After purification by nickel column, four trimer proteins were prepared. SDS-PAGE and Western blot were performed for protein identification. BALB/c mice were immunized at week 0 and week 3, and blood samples were collected to measure the activities of binding and neutralizing antibodies in serum.Results:SDS-PAGE and Western blot showed that the four proteins had stable trimer structure. Antigen-antibody affinity test showed that the four trimer proteins had strong affinity with RSV-specific monoclonal antibodies 8897, D25, Motavizumab, AM14 and Palivizumab. The titers of antibodies induced by the two T4 trimers were higher after the initial immunization, while there was a substantial increase in the titers of antibodies induced by Human codon-optimized trimer protein after the second immunization.Conclusions:PreF trimer protein can be prepared by adding any of the two different heterotrimer motifs, and induce effective binding and neutralizing antibodies in mice.
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BACKGROUND:Patella-patellar tendon junction (PPTJ) injuries are a common disease in athletes and vulnerable to re-injury. Post-injury training is a double-edged sword. OBJECTIVE:To examine the effect of post-injury resting days on the quality of healing in PPTJ in view of histomorphology, expression levels of growth factor and collagen, and muscle tension. METHODS:Forty New Zealand white rabbits were randomly divided into control, and five post-injury training groups representing different time to restart training after acute injury:the rabbits restarted training at 24 (PIT24), 48 (PIT48), 72 (PIT72), 96 (PIT96) and 120 (PIT120) hours, respectively. All post-injury training groups received acupuncture at the PPTJ and then underwent 4-week low-intensity training. Afterwards, all rabbits were killed to take specimens for hematoxylin-eosin, safranin O and immuohistochemical staining. The muscle tension and hardness were detected using Myoton-3. The thickness of fibrous cartilage zones, cell density, as well as the expression levels of vascular endothelial growth factor and collagen were measured on Metamorph software. RESULTS AND CONCLUSION:Compared with the control group, in the PIT groups, the collagens arranged in disorder, and decreased cell density, increased mucopolysaccharide andrising tidephenomenon appeared. The fibrocartilage in the PIT24 and PIT120 groups was significantly thicker than that in the control group (P<0.05). The ratio of collagen type Ⅰ/Ⅲ was significantly decreased in the PIT24 and PIT48 groups compared with the control group (P<0.05), and the highest in the PIT72 group. The expression level of vascular endothelial growth factor in the PIT48, PIT72 and PIT96 groups was significantly higher than that in the control group (P<0.05). The muscle tension was negatively correlated with the collagen ratio (r=-0.538;P<0.01) and chondrocyte density (r=-0.570;P<0.01), while muscle hardness was positively correlated with the expression level of vascular endothelial growth factor (r=0.613;P<0.01). Our results suggest that different resting time makes significant differences in the PPTJ healing and the 72-hour resting is the best choice. Moreover, muscular properties are related to the cel density, collagen synthesis and growth factor expression, which can be used as a non-invasive indicator to guide post-injury training and prevent recurrence.
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Objective To evaluate the effect rotation constancy static magnetic field(RCSMF) on the expression of bone morphogenetic protein-2 (BMP-2) and platelet-derived growth factors (PDGFs) in fracture healing, and elucidate the possible mechanism of RCSMF promoting fracture healing.Methods A total of 80 rats with femur closed fracture were randomly divided into a magnetic treatment group (M) and a control group (C).The M group was given RCSMF treatment 30 minutes per day, 6 times per week, using 0.4 T 6 Hz magnetic field, while the C group was not given any intervention.Femurs and calluses were subjected to BMP-2, PDGF-A and PDGF-B immunohistochemistry assay at 2, 3, 4 and 8 weeks after the treatment.The integrated optical density (IOD) of positive staining was calculated.Two-factor variance analysis was used to compare the main effect of time, treatment and that of their interaction.Results For both groups, the IOD of immunoreactivity of BMP-2 at 2, 3 and 4 weeks after the treatment was significantly higher than that 8 weeks after the treatment, and the value at four weeks after the intervention was also significantly higher than that at 2 weeks after it.There was a significant interaction effect of time × treatment of BMP-2 (F =3.17, P < 0.05).Significant differences were observed in the IOD of positive staining PDGF-A at different time points for both groups : the value at 2 weeks after treatment was significantly higher than that at 3, 4 and 8 weeks after the treatment, and that at 3 and 4 weeks after treatment was also significantly higher than that at 8 weeks after the treatment.The IOD of positive staining PDGF-B of the M group (57.6 ± 2.1) was significantly higher than that of the C group (50.11 ± 2.22, P < 0.05).There was significant difference in the IOD of positive staining PDGF-B (F =50.06 ,P < 0.01) at different time points : the IOD of positive staining PDGF-B at 3 weeks after treatment was significantly higher than that at 2 and 8 weeks group after the treatment, and the value at 2 and 4 weeks after treatment was also significantly higher that at 8 weeks after the treatment.Conclusion RCSMF may elevate the expression BMP-2 and PDGF-B to promote fracture healing.
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Objective To investigate the epidemic patterns and the characteristics of influenza in Chi-na through a Meta-analysis based on the studies published in domestic literatures.Methods Related articles published during 2005 to 2012 were screened out from domestic databases and analyzed through a Meta-analysis with Review Manager 5.0 software.Results Twenty-two articles covering 957 901 patients with influenza-like-illness (ILI) and 148 233 pathogen samples were screened out according to the inclusion criteria.No significant difference with the ILI diagnosis rate was found between subjects at age 0-4 years and those at age 15-59 years. Higher ILI diagnosis rates were observed in those two groups as compared with subjects elder than 60 years old. Most of the pathogen samples were carried by subjects aged 25-59 years.More influenza virus strains were isola-ted in 2009 as compared with those of the seven other years (OR=2.25, 95%CI=1.27-3.70).There was sta-tistical difference between the numbers of influenza A H1N1 and seasonal influenza A strains (OR=2.25, 95%CI=1.30-3.91) .Significant difference was also observed between the numbers of influenza A and influenza B strains (OR=4.05, 95%CI=2.53-6.47).Conclusion There was significant difference with the diagnosis rate between subjects aged 0-4 years and those aged≥60 years.More attention should be paid to people at high risk of infection (0-4 years old and≥60 years old) and those at 25-29 years with high mobility and social inter-course for the timely prevention and control of pandemic influenza.The detection rate of influenza virus strains was increased during the outbreak of novel influenza A H1N1 infection in 2009.After that outbreak, the detec-tion rate of novel influenza A H1N1 strains was 2.25 times the rate of seasonal influenza strains.The detection rate of influenza A was 4.05 times the rate of influenza B virus strains.Therefore, it is necessary to strengthen the surveillance for influenza A virus and other epidemic influenza virus strains.
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<p><b>OBJECTIVE</b>To investigate the effect of temperature on the stability of intermediate and final products of inactivated enterovirus 71 vaccine, which was prepared in human diploid cells.</p><p><b>METHODS</b>The different batches of harvest viral cultures, the vaccine stock solutions and the final productions of inactivated enterovirus 71 vaccine were stored at different temperatures. The samples of viral culture stored at -20°C or 4°C were harvested at 0, 6, 12 and 24 months later. The samples of vaccine stock solutions stored at -20°C were harvested at 0, 6, 12 and 24 months later, and that stored at 4°C were harvested at 0, 1, 3, 6 and 12 months later. The samples of finial products were harvested at different time points (0, 6, 12 and 24 months for storing at 4°C; 0, 7, 14, 28, 42 and 60 d for storing at 25°C; 0, 3, 7, 14 and 21 d for storing at 37°C). The viral titer, antigen content, antigen purity, endotoxin content, effectiveness, pH and appearance of samples were determined, respectively. A total of 1 800 BLAB/c mice were immunized by vaccine and 150 control mice were injected by diluents without antigen via intraperitoneal. The tail vein blood (500 µl per mouse) from 1 950 mice were harvested after 4 weeks post injected. The neutralization antibody titers of the serum were tested to calculate the half effective dose (ED50) of final products. All results were analyzed using analysis of variance to compare the differences of the above indexes.</p><p><b>RESULTS</b>The viral titers of harvest viral culture of inactivated EV71 vaccine were (6.67 ± 0.13), (6.56 ± 0.09), (6.52 ± 0.04), (6.39 ± 0.16) lgCCID50/ml (CCID50, the half cell culture infective dose) after 0, 6, 12 and 24 months storage at -20°C; and (6.67 ± 0.13), (6.41 ± 0.13), (6.19 ± 0.18), (5.97 ± 0.09) lgCCID50/ml at 4°C. The viral titers reduced with time (F = 9.81 or 44.16, P < 0.05). The antigen contents of the vaccine stock solution were maintained at (3 626.67 ± 1 382.56) EU/ml within 3 months at 4°C, but were (2 080.00 ± 876.36), (951.17 ± 346.35) EU/ml at 6 and 12 months, respectively. The ED50 of the final production were (31.00 ± 2.71), (32.93 ± 3.22), (39.37 ± 3.44) and (46.04 ± 3.25) EU/ml after 0, 6, 12 and 24 months storage at 4 °C, but were (31.00 ± 2.71), (32.23 ± 2.66), (34.70 ± 1.77), (40.04 ± 2.10), (47.78 ± 1.93) and (56.97 ± 0.50) EU/ml at 0, 7, 14, 28, 42 and 60 days at 25°C, and were (31.00 ± 0.00), (36.20 ± 0.00), (41.87 ± 0.50), (53.25 ± 0.50) and (64.84 ± 0.58) EU/ml at 0, 3, 7, 14 and 21 days at 37°C, respectively. The ED50 had increased with the time by and had significantly differences compared with the beginning level (F = 28.49, 215.15 or 156.12, P < 0.05).</p><p><b>CONCLUSION</b>There is a good stability of the intermediate and final productions of inactivated enterovirus 71 (EV71) vaccines, within 24 months at -20°C or 6 months at 4°C storage for viral culture, 24 months at -20°C or 3 months at 4°C storage for stock solution and 24 months at 4°C or 28 d at 25°C or 7 d at 37°C storage for finial vaccine.</p>
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Animals , Humans , Mice , Drug Storage , Methods , Enterovirus A, Human , Immunization , Vaccination , Vaccine Potency , Vaccines, InactivatedABSTRACT
Objective To compare the clinical efficacy of targeting arterial embolization before sacral chordoma excision and temporary balloon oc-clusion of abdominal aorta during sacral tumors surgery. Methods A total of 34 patients with sacral chordoma were recruited for this study and fol-lowed up. Totally 18 patients received bi-iliac artery,sacrococcygeal artery and other target arteryl embolization before excision(target arterial emboli-zation group),and the other 16 received temporary balloon occlusion of abdominal aorta during surgery(balloon occlusion group). The blood loss during operation,drained blood after operation,operation time and wound healing were recorded. Results The blood loss during operation,drained blood after operation and operation time in targeting arterial embolization group were less than balloon occlusion group. There was no statistical differ-ence in wound healing between the two groups. Conclusion Targeting arterial embolization can be more effective than temporary balloon occlusion of abdominal aorta in the control of blood loss.
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Objective To compare the therapeutic efficacy of 125Iodine seed implantation and gamma knife in treating portal vein tumor thrombosis (PVTT) in hepatocellular carcinoma (HCC), and to discuss their clinical applications. Methods Between January 2009 and May 2013, a total of 60 HCC patients associated with PVTT were encountered at authors’ hospital. The patients were divided into gamma knife group (n=30) and 125Iodine seed group (n = 30). Gamma knife treatment was carried out for the patients of gamma knife group. A total of 3 - 10 target points were designed. Single dose of 3 - 6 Gy per fraction was used and 2 - 5 times radiotherapy every week were conducted. The total treatment dose was 30 - 50 Gy. In designing the therapeutic scheme, 50%-70%of iso-dose curve contained PTV. 125Iodine seed implantation was performed for the patients of 125Iodine seed group. After the treatment, the change of PVTT size was assessed. The clinical efficacy was evaluated. Results Three months after the treatment, in gamma knife group complete remission (CR) was seen in 0 patient (0%), partial remission (PR) in 8 patient (26.6%), stable disease (SD) in 17 patients (56.7%) and progressive disease (PD) in 5 patients (16.7%). In 125Iodine seed group, CR was obtained in 3 patients (10.0%), PR in 19 patients (63.4%), SD in 7 patients (23.3%) and PD in one patient (3.3%). Statistically significant differences in responding the therapy existed between the two groups (P < 0.05), although no significant difference in the improvement of ascites and hepatic function existed between the two groups (P<0.05). Conclusion For the treatment of portal vein tumor thrombosis of hepatocellular carcinoma, the short-term efficacy of 125Iodine seed implantation is much better than that of gamma knife, although its long-term effect needs to be further studied.
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Objective To discuss the comparability of blood urea nitrogen (BUN),creatinine (Cr),uric acid (UA) results and provide basis for result concordance in different biochemical systems.Methods The within-run precision and the between-day precision for the median and high values of quality control materials were tested in Beckman LX-20 and Hitachi 7600 chemistry analyzer respectively,and the evaluation criteria were according to the precision requirement of manufacturer and our laboratory.According to American Clinical and Laboratory Standards Institute (CLSI) document EP9-A2,serum BUN,Cr,UA from 40 patients were detected by Hitachi 7600 chemistry analyzer as experimental method and by Beckman LX-20 chemistry analyzer as comparative method.The relative bias ( SE%) between experimental method and comparative method was calculated and the concordance of biochemical analysis system was determined according to EQA requirements and acceptable performance criteria Results The between-run and between-day precision coefficients of variation for BUN,Cr and UA in two biochemical analysis systems were lower than the precision requirement of manufacturer and our laboratory.In Hitachi 7600 chemistry analyzer,the SE% for BUN at three levels of medical decision were 9.4%,2.0% and 0.8%,the SE% for Cr were 7.3%,3.8% and 2.8%,the SE% for UA were 7.9%,1.7% and 1.2%.Two biochemical analysis systems were not comparable in BUN analysis because SE% of serum BUN levels detected by Hitachi 7600 at low medical decision level exceeded the specified range.The remaining items between two biochemical analysis systems were concordant.Conclusion While the same test is detected by more than two systems in one laboratory,it is necessary to do method comparison and bias evaluation for concordance in order to ensure the comparability among test results.
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In order to understand the roles of the other subunits, we investigated expression of the NMDA receptor subunits (NR2A and NR2B) in visual cortex of normal and anisometropic amblyopia kittens with different ages in the present study. We examined the expressions of NR2A and NR2B in the visual cortex of the kittens by immunohistochemistry with polyclonal anti-NR2A antibody and anti-NR2B antibody, respectively. Using immunohisto-chemical Streptavidin Perosidase (SP) method, we observed the dynamic changes of NR2A and NR2B with microscope and computer-assisted image analyses. We found that NR2A and NR2B remained low expression after the peak of the critical period of kitten visual development; compared with normal group of the same age, NR2A expresses low. However, the difference is not significant for NR2B before maturation period of visual development. NR2B rises after the maturation period of visual development. According to this, the component of NR2A and NR2B can be affected by anisometropia. This research suggests that the difference of NR2A and NR2B expressions may affect the formation of amblyopia.
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Animals , Cats , Female , Male , Amblyopia , Metabolism , Receptors, N-Methyl-D-Aspartate , Genetics , Metabolism , Vision, Ocular , Physiology , Visual Cortex , MetabolismABSTRACT
Objective To make a preliminary evaluation of the degradation of AZ31 bioabsorbable magnesium alloy stent implanted in the abdominal aorta of experimental rabbits.Methods Twelve AZ31 biodegradable magnesium alloy stents were separately deployed in the infrarenal abdominal aortas of twelve New Zealand white rabbits.Every three experimental rabbits were sacrificed each time at one,two,three and four months after the procedure of stenting.The stenting segment of the aortas were harvested,radiographod and sent for pathologic examination to observe the degradable performance of the stent.Results All animals survived form the operation in the scheduled follow-up period.Radiographically and pathologically,the stents were fully expanded with perfect shape one month after the procedure,and part of the stent struts began to be degraded and fractured in two months,resulting in the loss of its supporting function.Three months after the implantation most stents were corroded.and in four months all the stents become completely destroyed.The estimating time for producing complete degradation of AZ31 magnesium alloy stents in rabbit's aorta was 104.5 days.Conclusion AZ31 bioabsorbable magnesium alloy stents implanted in rabbit abdominal aorta will lose their radial force in two months.How to prolong the functioning time of the implanted stents is the next research target.
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Objective To distinguish residual tumor from inflammation after radiofrequency ablation (RA) for hepatic VX2 carcinoma in rabbits according to the comparative study between CT and pathological findings.Methods CT and pathologic examination were performed in different stages of RFA for rabbits hepatic VX2 models,and their different performances were observed.Results Marginal enhancement band was showed with enhanced CT of both residual tumor and inflammation.Moreover,liver tissues peripheral to enhancement band were in gradual weaken pattern.The enhancement band of inflammation was most obvious on the 2~(nd) day after RFA,but weakened gradually and disappeared two weeks later.Conclusion The residual tumor and inflammation could not be distinguished through enhanced CT scanning within 1 week after RFA.Low intensity lesions with peripheral enhancement 2 weeks after RFA should be recognized as residual tumor.
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Objective To evaluate the factors which might affect the successful reopening of occluded subclavian artery. Methods During the period of Jan. 1998-March 2007, endovascular stent placement was performed in 20 patients with occluded subclavian artery, including 12 males and 8 females, aged 17-74 years (mean 53 years). The procedures were carried out by using super-slippery guide wire via anterograde or bilateral access. Results Successful reopening of occluded subclavian artery was obtained in 14 cases and the treatment ended in failure in 6 cases. In 14 patients with successful results, the mean length of diseased artery was 3.00 cm and the mean course of disease was 9.83 months, which were 2.13 cm and 27.6 months respectively in 6 cases of failure. The successful reopening rate was 80% in arthrosclerosis cases, while it was 50% in aorto-arteritis obliterans. The successful reopening rate was 100% in those cases whose occluded artery showing a sharp stump, while the successful reopening rate was 33% in those cases whose occluded artery showing a round stump, with a statistically significant difference between the two (P< 0.05). The successful rate in cases performed via anterograde access was 65%, and it was only 43% in cases performed via bilateral access. Conclusion The etiology, the duration of disease and the shape of stump bear a close relationship to the successful reopening rate, while no obvious correlation exists between the length of diseased artery and the reopening rate. Higher reopening rate can be achieved when the procedure is performed via anterograde access.