Objective To investigate localization and co-existence of the tyrosine hydroxylase (TH)and calcitonin gene-related peptide(CGRP)on the cochlear spiral modiolar artery(SMA) and its branches.Methods The SMA was dissected from the modiolus so that the entire length of the vessel and many of its branches could be observed. Immunohistochemical ABC method and confocal immunoflurescence technique were employed to localize each compound and to determine whether the TH and CGRP coexist in neurons of the SMA.Results Microscopic examination of whole vessel preparation revealed numerous TH and CGRP-positive neural networks innervating the SMA and its branches.The labeled nerve fibers showed distinct arborization ,varicosities and overlap,and were of different diameters. The overall distribution of neuronal labeling suggested a gradual decrease of innervation density from proximal (cochlear base)to more distal vessels, i.e., apical cochlear vessels and lateral radiating arterioles. Confocal immunofluorescence microscopy of double-labeled TH and CGRP neurons showed that a number of the TH-and CGRP-positive neurons were co-labeled. Thus, TH and CGRP partially co-exist within the neuronal innervation of SMA.Conclusion These findings support a hypothesis that specific neuropeptide and adrenergic neurons regulate cochlear blood flow.

Objective To explore an alternative strategy of ganciclovir-mediated cytotoxicity and study the mechanism of enhancing bystander effect of HSV-TK/GCV system by combination gene transfer of Rb gene and HSV-TK gene.Methods Recombinant eukaryotic expression plasmid pcDNA3.1-Rb harbour ing1.65kb of Rb gene was constructed.The pcDNA3.1-Rb and the plasmids tgCMV/HyTK car-ry ing HSV-TK gene were transfected respectively or co-transfected into the osteosarcoma cell line OS732by lipofection using DOTAP reagent.The mRNA expression of Rb gene and HSV-TK gene were detected by RT-PCR and Northern blot.Cell counting,cell cycle analysis and soft agar colony formation test were adopted to observe cell growth features.The expression of gap junction Connexin43gene was performed by RT -PCR and Western blot.The direct confirmation of gap junction intercellular commu ni cation was demonstrated by Lucifer yellow dye transfer.Cell sensitivity to GCV and″bystander effect″of HSV-TK/GCV system were measured by MTT assay.Results The eukaryotic expression plasmid pcD NA3.1-Rb was constructed successfully.The transfected cell lines OS732TK,OS732Rb and OS732RbTK were har-vested.mRNA expression of HSV-TK gene was demonstrated in OS732TK and OS732RbTK cells.Both exogenous and endogenous Rb gene expression could be detected in OS732Rb and OS732RbTK cells si-multa neously.Compared with parental cell OS732,OS732Rb and OS732RbTK cells changed their mor -phology and decreased the growth rate;the ability of soft agar colony formation reduced and the cell cycles were arrested at G 0 G 1 checkpoint.The Connexin43expression and gap junction in tercellular communica-tion en hanced in OS732Rb cell.GCV was of toxicity to only TK -positive cells,OS732TK and OS732RbTK,in a concentration dependent manner,the mixed coculture experiments of OS732and OS732Rb with the TK-negative cell,both showed sensitive to GCV,but the survival rate of OS732Rb cell was significantly lower than OS732cell under the same condition.Conclusion Coordinate ex pression of Rb andHSV-TK gene conferred a more po tent bystander effect by enhancing gap junction intercellular communica-tion and in hibiting prolifera tion.

Objective To study the initial and the follow-up manifestations of infants with Langerhans’ cell histiocytosis (LCH) and evaluate the relationship between X-ray manifestations and clinical prognosis. Methods The radiological manifestations were analysed in 22 cases of LCH proved by clinical features, laboratory data, and pathology. According to Lavin’s Grade, the 22 cases were classified as Grade I (9 cases), Grade II (9 cases), and Grade III (4 cases). Plain radiography was performed in all cases during the initial and the follow-up,including cranial bones, chest, long bones, and spine. CT was performed in 12 cases. Results Of the 22 cases, there were 7 cases which lesions were limited to bones. 5 of them were demonstrated with ossification and remodeling of the involved bone after chemotherapy.The remaining 15 cases were associated with multi-systemic involvements, only 4 of which improved after chemotherapy. Among these 15 cases, 7 had lung involvement, only 2 of which had gotten better. Hypothalamus/pituitary axis was affected in 1 case. Conclusion The extension of the bone involvement can reflect LCH clinical prognosis. However,the lung involvement of LCH, which mainly presented with interstitial lesion, is not sensitive for the evaluation of clinical situation.LCH with limited bone destructions have a better prognosis than those with the multi-systemic involvements.

Animals ; Diazepam ; pharmacology ; Drug Antagonism ; Electroencephalography ; Female ; Male ; Pyrazoles ; poisoning ; toxicity ; Rats ; Rats, Sprague-Dawley

Aconitum ; poisoning ; Animals ; Disease Models, Animal ; Female ; Hemoperfusion ; methods ; Male ; Plant Poisoning ; therapy ; Rabbits

Objective To isolate specific humanized anti-D-dimer scFv(single chain Fv) antibody from scFv phage libraries. Methods Isolate anti-D-dimer positive clones from Tomlinson I + J phage libraries by three rounds of panuing, then sequence monoclonal genes by bideoxy-mediated chain termination and express soluble scFv antibody; Pick out anti-D-dimer antibodies with high specificity and affinity by ELISA.Results After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer scFv fragments were selected. By polyclonal and monoclonal phage ELISA and gene sequencing, 20 different full-length monoclonal scFv phages were identified, the result of soluble scFv ELISA showed that 20 full-length monoclonal scFv were expressed smoothly. According to the result of soluble scFv ELISA, in 5 scFv antibodies with high value of A450 selected, 3 scFv antibody fragments showed high specific and affinity. Conclusion Antibody phage display was an effective, rapid method to isolate anti-D-dimer antibodies with high specificity and affinity.

AIM: To investigate killing effect of herpes simplex virus thymidine kinase/ganciclovior system(HSV-TK/GCV) on osteosarcoma cell and its mechanisms.METHODS: Recombinant retroviral vector (DORHyTK) containing hygromycin phosphotransferase-thymidine kinase fusion gene(HyTK) was constructed and introduced into human osteosarcoma cell line OS732 with DOTAP. DNA and total RNA extracted from HyTK expressing cells (OS732TK) were tested by PCR and RNA dot blot analysis. A chemosensitivity of OS732TK cells to GCV and “bystander effect” were measured by means of MTT colorimetric assay. Hoeschst 332258 staining and flow cytometric analysis were performed for mechanism of HSV-TK/GCV system gene therapy. RESULTS: DORHyTK was constructed successfully; the HyTK gene existed and expressed in OS732TK cells; All OS732TK cells were killed after 5 days of exposure to 5 mg/L GCV. The “bystander effect” was observed in both a high population and a low population, but the former was stronger than the latter. Hoeschst 332258 staining revealed the characteristic hallmarks of apoptosis, however necrosis also existed. Flow cytometric analysis of DNA content showed a G0-G1 phase blockade. CONCLUSION: HSV-TK/GCV gene therapy system showed its strong killing effects on osteosarcoma cells OS732; The phenomenon of “bystander effect” was also very apparent. GCV exposure induced both necrotic and apoptotic death in HSV-TK expressing cells, and perturbed the cell cycle. HSV-TK/GCV gene therapy system may provide a new therapeutic approach for treatment of osteosarcoma.

Purpose　To observe the killing effect of HSV-TK/GCV system on osteosarcoma cells. Methods　The eukaryotic exprssoin vector (tgCMV-HyTK) containing HSV-TK gene was transduced into human osteosarcoma cell lines SAOS-2 by DOTAP. Total RNA extracted from HSV-TK expressing cells (SAOS-2TK) were tested by RNA dot blot analysis. A chemosensitivity of SAOS-2TK cells to GCV and “bystander effect” were measured by means of MTT colrimetric assay. Results　HSV-TK gene was expressed in SAOS-2TK cells. SAOS-2TK cells were susceptible to the cytotoxicity of GCV. A meaningful “bystander effect” was demonstrated. Conclusion　HSV-TK/GCV system may kill selectively the human osteosarcoma cell line SAOS-2.

Objective Frontal resting EEG asymmetry was used to evaluate the gender effect of emotion regulation.Methods The resting EEG was recorded in 30 healthy volunteers (19-26 years old) by the guide 40 amplifier,and off-line analysis on EEG data were conducted by scan4.3 software.The statistical analysis were performed using SPSS 17.0.Results In normal human,frontal resting EEG alpha power was statistically significant(F=4.918,P =0.035).Asymmetry and gender interaction group (F =0.668,P =0.421),electrode point and gender interaction group (F =0.283,P =0.756),electrode point and asymmetry interaction group (F =1.418,P =0.260),electrode point,asymmetry and gender interaction group (F =1.164,P =0.327) had no statistical significance (P ＞ 0.05).The comparison of male and female asymmetry degree of each electrode point (F4/F3 group t =0.465,F8/F7 groupt=0.809,FP2/FP1 group t=1.542) had no statistical significance (P＞0.05).Conclusion Normal human brain hemispheres exist asymmetry.The asymmetry does not exists gender effect.In non-task state,frontal resting EEG alpha power asymmetry can not be recognized as the indicator of emotion regulation ability.

Objective:To discuss the role of incision for short-term outcomes of simultaneous resection in synchronous colorec-tal liver metastases (sCRLM). Methods:We reviewed the data of 37 patients who underwent simultaneous resection between January 2009 and December 2014 in our department and compared the short-term outcomes between Mercedes and midline incisions. Results:Mercedes and midline incisions were used in 19 and 18 patients, respectively. The two groups showed similarities in patient characteris-tics, major hepatectomy, surgery time, blood loss, and hilar block time. The midline group comprised more rectal cancer patients (P<0.001). The two groups did not differ significantly in complication incidence (47.4%vs. 16.7%, P=0.08) and postoperative stay time (22.1 ± 9.5 d vs. 17.2 ± 6.7 d, P=0.08). At body mass index (BMI)<25, the complication incidence (P=0.046) and postoperative stay time (P=0.051) were lower in the midline group than in the Mercedes group. Conclusion:Midline incision provided similar exposure in simultaneous resection for sCRLM and was better than Mercedes incision in rectal cancer patients. Patients with midline incision may attain better short-term outcomes if BMI is<25.