Objective To explore the effects of hypoxic post-conditioning on cognitive function and the expression of silent information regulator 1 (SIRT1) in hippocampal CA1 of rats with cerebral ischemia. Methods Sixty Sprague-Dawley rats were randomly divided into sham operation group, model group and treatment group with 20 cases in each group. Each group was divided into one day, two days, three days, seven days subgroups according to the time of ischemia reperfusion. Global cerebral ischemia reperfusion was induced with modified Pulsinelli′4-vessel occlusion. The treatment group received 8%oxygen for two hours after ischemia. The cognitive function was assessed with Morris water maze test. Morphological changes of the hippocampal CA1 region were observed by HE staining. The expression of SIRT1 in the hippocampal CA1 region was detected with immunohistochemical assay and Western blotting. Results Compared with the model group, the escape latency significantly shortened (P<0.05), the number of times crossing the platform increased (P<0.05), the speed and the percentage of time spent in the platform quadrant increased (P<0.05), and the total distance decreased (P<0.05);the expression of SIRT1 in hippocampal CA1 increased (P<0.05) and the number of normal neurons increased (P<0.05) in the treatment group. Conclusion Hypoxic post-conditioning can improve the cognitive function of rats with global cerebral ischemia, which may relate with up-regulating SIRT1 in hippocampus.

Objective To predict and identify T cell epitopes of major group 3 allergen derived from Dermatophagoides fari-na(Der f 3). Methods The T cell epitopes of Der f 3 were analyzed through the sequence analysis by using the bioinformatics online tools. The five predicted peptides of T-cell epitopes were artificially synthesized. The spleen lymphocytes were co-cultured with the five T cell epitopes by using the modified MTT method and the levels of IL-2,IFN-γ,IL-4 and IL-5 in the supernatant of the cultures were detected by ELISA. Results Five T cell epitopes of Der f 3 were predicted and three of which could pro-mote the proliferation of the mouse spleen lymphocytes. The secretions of IL-2 and IFN-γwere significantly induced and the se-cretions of IL-4 and IL-5 were significantly decreased by three of five prediction epitopes of Der f 3:37GDCPYQISLQSSSHFC-GG54,98IYQHENYDSMTIDNDVALIKLKTPMT123 and 164SELQRVDIDVVSREQCDQLYS184. Conclusion Three T cell epitopes of Der f 3 have been initially identified,which lays the foundation of the diagnosis and treatment of asthma.

Objective To develop a portable pressure detector to facilitate the battlefield self and buddy aids training for dressing,hemostasis and fixation.Methods The changes of pressure were converted into the ones of electric current with the pneumatic cuff,catheter and membrane pressure sensor,and then transmitted to the panel display by Bluetooth.The efficacy for the training was determined based on the acquired data.Results The detector implemented quantifying of the pressures during dressing,hemostasis and fixation,and non-medical staff obtained the results of battlefield treatment training easily to execute rapid assessment of battlefield self and buddy aids training.Conclusion The device gains advantages in visualized data,portability,easy operation and accurate measurement,and contributes to battlefield self and buddy aids training.

Objective To establish a treatment proposal of thyroid adenoma by using percutaneous radiofrequency ablation(RFA) and investigate its techniques and skills, means and steps, and safety and efficacy. Methods Contrast-enhanced ultrasound-guided percutaneous RFA of thyroid adenomas were conducted on 202 patients by using an auto-controlled bi-polar electrode system. The indications of thyroid RFA,the optimal puncture route,the ways of anesthesia administration, protection of vital neck vessels and recurrent laryngeal nerve(RLN) and reduction of bleeding from core biopsy, indicators of ending ablation procedure following a complete ablation were investigated and analyzed. Resalts An adenoma smaller than 20 mm in maximal diameter was the optimal candidate for RFA. Either of two puncture routes could be selected upon the target lesion's location. Areas surrounding to the thyroid capsule needed adequate local anesthesia to kill pain. Liquid-isolating maneuver could effectively protect carotid artery and RLN from core needle cutting and electrode heating injury. Advanced block of supplying arteries with heating markedly reduced bleeding involved in the biopsy. Multipoint and multicenter ablation was essential to a complete coagulation. Filling-defect in the ablated adenoma on CEUS was the key sign to terminate ablation procedure. Conclusions Percutaneous bi-polar RFA was proved feasible, effective, safe and supermicroinvasive for treating thyroid adenoma under the way stated here of puncture and technical points and use of CEUS for monitoring.

Objective To predict and identify the linear B-cell epitopes in the major group 3 allergen derived from Derma-tophagoides farina(Der f 3). Methods The linear B-cell epitopes of Der f 3 allergen were analyzed based on the physicochemi-cal properties of amino acids including antigenicity,surface accessibility,flexibility,hydrophilicity,beta-turn by online bioinfor-matics softwares. The eight predicted peptides of linear B-cell epitopes were artificially synthesized and incubated with three aller-gic serum pools(4 serum samples in each),which were consisted of total 12 serum samples from the allergic individuals,and the strong positive epitopes were selected. Results Eight B-cell epitopes from Der f 3 were predicted successfully. Five of eight B-cell epitopes were identified with strong IgE-binding abilities followed by specific IgE assay. The amino acid sequences of them were following:33KAKAGDCP40, 86HASGGEKIQVAEIYQHENYDSMTID110, 118LKTPMTLDQTNAKPVPLPPQGSDVKVG144, 156QEGSYSLP163 and 199DVANGGVDSCQGDSGGPVVD218. Conclusions Five linear B-cell epitopes of Der f 3 allergen have been identified successfully. This result might provide a basis of the diagnosis and treatment for asthma.

Objective To study the induction of integrin molecules mediated by CYR61 in human peripheral blood mononuclear cells(PBMC). Methods A recombinant expression plasmid containing full length of human CYR61 labeled with human IgG Fc fragment was constructed and identified by DNA sequencing.COS7 was used as host cell for identification of secretary CYR61 expression,confirmed by Western blot method.The commercial lipofectin was adopted for recombinant plasmid transfection into PBMC.Real-time PCR was ultilized to analyze expression patterns of CYR61 and integrin molecules in transfected PBMC. Results The insert sequence was correct in the recombinant plasmid.Western blot test showed that CYR61 protein secreted into culture supernatant or was in COS7 cytoplasm.The recombinant plasmid was transfected into PBMC stimulated with phytohemagglutinine(PHA) to induce CYR61 expression and secretion.The results demonstrated that exogenous CYR61 transcribed rapidly after being incubated with PHA and reached the peak after 24 h.But the expression dropped down quickly to a very low level after 48 h.Simultaneously,integrin molecules expressed just after CYR61 transcription.In the set of integrin molecules tested in the study,?v,?M,?3 and ?5 expression were higher than the other integrin molecules(P

Objective To investigate the microstructural abnormalities of cerebral white matter in patients with medication-overuse headache with diffusion tensor imaging.Methods Diffusion tensor magnetic resonance imaging (DT-MRI) was carried out in 80 migraine patients with medication-overuse headache (case group) and 80 age-and sex-matched healthy controls (control group).Fractional anisotropy (FA) and apparent diffusion coefficient (ADC) were measured and the correlation between clinical characteristics and the changes was examined.Results ① The FA values at the bilateral orbitofrontal cortex,cingulate cortex,splenium of the corpus callosum and the right anterior limb of the capsula interna were significantly lower than those in the controls(P＜0.05).② The ADC values at the right orbitofrontal cortex,left inferior frontal gyrus and anterior cingulate cortex were significantly higher than those in the controls (P＜0.05).③ There were negative correlations between FA values at the bilateral orbitofrontal cortex and right hind legs of the capsula interna and headache course as well as frequency.There was negative correlation between FA values at the left hind legs of capsula interna and headache frequency.④ There were positive correlations between ADC values at the left orbitofrontal cortex and headache course as well as frequency.There were positive correlations between ADC values at the right orbitofrontal cortex and prior cingulate cortex and headache frequency.There was positive correlation between ADC values at the posterior cingulate cortex and headache course.Conclutions There might be an integrity change of neurofibrotic microstructures in the bilateral orbitofrontal cortex and cingulate cortex in patients with medication-overuse headache,FA values are negatively associated with headache course and frequency whereas ADC values are positively associated with headache course and frequency.

Objective:To investigate the distribution of 99Tc m-methylene diphosphonate (MDP) at different stages of bone injury repair. Methods:A total of 30 rabbit models of femur injury were established by the method of electric drill and perforation of femur. According to the different stages of bone injury repair (at 1, 2 and 3 week), rabbits were divided into group A, B and C ( n=10 each group). Femoral SPECT/CT imaging was performed on the last day of different stages of bone injury repair to obtain radioactivity counts in the region of interest (ROI) on the test side and control side and to calculate target/background ratio (T/B). The light intensity of 3 groups was analyzed by phosphor screen imaging and the distribution of 99Tc m-MDP in bone cells was observed by autoradiography. One-way analysis of variance and paired t test were used to analyze the data. Results:The T/B values of group A, B and C were 1.16±0.14, 1.39±0.23 and 1.18±0.10, respectively ( F=5.83, P<0.01). There were significant differences of the maximum radiation count between the test side (50.00±12.45, 59.50±12.83 and 55.10±9.26) and the control side (43.20±9.57, 50.00±12.30 and 44.30± 6.50) in group A, B and C ( t values: 3.24, 2.28 and 5.77, all P<0.05). There were significant differences in the light intensity of bone specimens in group A, B and C by phosphor screen imaging (37 324.67±6 481.50, 60 950.33±9 781.72 and 43 905.00±4 957.92; F=8.25, P=0.02). 99Tc m-MDP were deposited in both intracellular and extracellular during different stages of bone repair in osteocytes and osteoblasts under autoradiography. Conclusion:At different stages of bone injury repair, the concentration of 99Tc m-MDP is significantly distributed, suggesting that there are other ways of concentration mechanism of 99Tc m-MDP in bone tissue besides the chemical adsorption with hydroxyapatite.

OBJECTIVETo construct a vector encoding T-cell epitopes of major allergen group 1 of Dermatophagoides pteronyssinus as a vaccine delivered by MHC class II pathway.

METHODSThe nucleotide sequences of the 3 target genes were synthesized, including TAT, IhC and the recombinant fragment of Der p 1 encoding 3 T-cell epitopes. After amplification of the 3 target fragments by PCR and digestion with corresponding restriction endonucleases, the recombinant gene TAT-IhC-Der p 1-3T was ligated using T4 DNA ligase and inserted into the prokaryotic expression vector pET28a(+) to construct the recombinant plasmid pET-28a(+)-TAT-IhC-Der p 1-3T, which was confirmed by digestion with restriction endonucleases and sequencing. The recombinant vector was transformed into E. coli strain BL21 (DE3) and induced with IPTG, and the induced protein TAT-IhC-Der p 1-3T was detected by SDS-PAGE. After purification, the recombinant protein was confirmed by Western blotting and its allergenicity tested using IgE-binding assay.

RESULTSThe recombinant plasmid pET-28a-TAT-IhC-Der p 1-3T was successfully constructed as confirmed by restriction endonuclease digestion and sequencing and the expression of the recombinant protein TAT-IhC-Der p 1-3T was induced in E. coli. Western blotting verified successfull purification of the target protein, which showed a stronger IgE-binding ability than Der p 1.

CONCLUSIONWe successfully constructed a recombinant expression vector pET-28a-TAT-IhC-Der p 1-3T expressing a T-cell epitope vaccine delivered by MHC II pathway with strong IgE-binding ability, which provides a basis for further study on specific immunotherapy via MHC class II pathway.

Allergens ; immunology ; Animals ; Antigens, Dermatophagoides ; immunology ; Arthropod Proteins ; immunology ; Base Sequence ; Cloning, Molecular ; Cysteine Endopeptidases ; immunology ; Dermatophagoides pteronyssinus ; Epitopes, T-Lymphocyte ; Escherichia coli ; Gene Expression ; Genes, MHC Class II ; Genetic Vectors ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Vaccines ; immunology

High mobility group A2 protein (HMGA2), an architectural factor, is highly expressed in various cancer types including lung cancers. It is a candidate target for cancer therapy. RNAi is an effective gene silencing method with low cost and less time-consuming. It is possible to exploit this technology in therapy. Here, 5 siRNAs targeting Hmga2 gene (HMGA2 siRNA1-5) were designed and synthesized. MTT assay, colony formation assay, transwell assay and flow cytometry were used to evaluate the effects of these siRNAs on lung cancer cell lines (NCI-H446 and A549). Results from cell proliferation, clone formation, migration and apoptosis showed that HMGA2 siRNA1, 3, 5 could affect these aspects for both lung cancer cell lines. Among the five siRNAs, HMGA2 siRNA5 showed the greatest inhibition effects. The inhibition effects of HMGA2 siRNA5 are sequence specific and are not due to the induction of interferon response. Taken together, siRNAs targeting Hmga2 gene are potential candidates for lung cancer gene therapy.
Apoptosis ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colony-Forming Units Assay ; Gene Silencing ; Genetic Therapy ; HMGA2 Protein ; genetics ; metabolism ; Humans ; Interferons ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Point Mutation ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection