The human brain-derived neurotrophic factor (hBDNF) gene was cloned by polymerase chain reaction and the recombinant adeno-associated viral vector inserted with hBDNF gene (AAV-hBDNF) was constructed. Cultured rat hippocampal neurons were treated with Abeta(25-35) and serued as the experimental Abeta-induced neuronal damage model (AD model), and the AD model was infected with AAV-hBDNF to explore neuroprotective effects of expression of BDNF. Cell viability was assayed by MTT. The expression of bcl-2 anti-apoptosis protein was detected by immunocytochemical staining. The change of intracellular free Ca ion ([Ca2+]i) was measured by laser scanning confocal microscopy. The results showed that BDNF had protective effects against A-induced neuronal damage. The expression of the bcl-2 anti-apoptosis protein was raised significantly and the balance of [Ca2+]i was maintained in the AAv-hBDNF treatment group as compared with AD model group. These data suggested that recombinant AAV mediated a stable expression of hBDNF in cultured hippocampal neurons and resulted in significant neuron protective effects in AD model. The BDNF may reduce neuron apoptosis through increasing the expression of the bcl-2 anti-apoptosis protein and inhibiting intracellular calcium overload. The viral vector-mediated gene expression of BDNF may pave the way of a novel therapeutic strategy for the treatment of neurodegenerative diseases such as Alzheimer's disease.

Objective To compare the growth of three different cancer cell lines on chick chorioallantoic membrane (CAM), to select the best transplanted cancer cell line for establishing a transplanted tumor model and to observe the biological characteristics.Methods The human lung cancer cell line A549, human tongue cancer cell line TCA8113 and human liver cancer cell line QGY7703 were respectively inoculated into CAM at the 7th day of age.The chick embryo survival rate, tumor survival rate, tumor formation rate and induced angiogenesis were detected and the growth characteristics of the transplanted tumor model were observed.Results Compared with the groups inoculated with A549 cells and QGY7703 cells, the tumor formation rate of TCA8113 cells was the highest (P < 0.05), to be the best cancer cell line for transplanted tumor.The optimal inoculated number of cells was 8.0×106/chick embryo, the optimal growth period of the tumor was 4~8 d, and the best experiment time was 7 d after inoculation.Conclusion The TCA-CAM transplanted tumor model of tongue squamous cell cancer is successfully established for further study of the biological characteristics and mechanisms of tumor growth, angiogenesis, invasion and metastasis, and provide a good experimental animal model for anti-tumor drug screening.

Objective To study CYP11B2 mRNA and aldosterone secretion alteration in human adrenocortical carcinoma H295R cell after angiotensin Ⅱ (AT-Ⅱ) and potassium chloride stimulation,and to investigate the effect of adrenocorticotropic hormone receptor (ACTHR) on them.Methods Lentiviral vector was used to increase ACTHR expression.It was transfected into the H295R cells.Similarly,another H295R cells,without ACTHR vector,was used as the control group.ACTHR alteration was measured by Western blot and real-time polymerase chain reaction (RT-PCR).CYP11B2 mRNA was detected at 24 hours after 100 nmol/L AT-Ⅱ/16 mmol/L KCL stimulation,and the amplification of the two groups was compared.Aldosterone was measured by ELISA kit.Results Compared with those in control ceils,the protein and mRNA level of ACTHR in experimental cells were increased 2.4 times and 18 times respectively (P＜0.05).CYP11B2 mRNA of experimental cells was 1.7 times higher than control cells after 24 h stimulation of AT-Ⅱ.Aldosterone production was 121.98+8.31 and 104.05+6.88 ng/L respectively.The former amplification was 2.06 times higher than that of the latter (P＜0.05).Similarly,CYP11B2 mRNA of experimental cells was 19.2 times higher than control cells after 24 h stimulation of KCL.Aldosterone production was 137.67±10.35 and 104.05 ± 6.88 ng/L respectively.The former amplification was 3.13 times higher than that of the latter (P＜0.05).Conclusion Overexpression of ACTHR increases the sensitivity and response of CYP11B2 mRNA and aldosterone to AT-Ⅱ and KCL stimulation,and ACTHR is expected to become a key protein in understanding primary aldosteronism.

Objective To observe the effects of rehabilitation in the 55 prelingually deaf pediatric cases for two years after cochlear implantation ,factors including cochlear implantation and recovery time ;to compare the re-covery effects in the group of 1 to 3 years old children with the group of 3~5 years old (including the age of 3 years old) who lost their hearing before learning to speak ,and to provide clinical evidence for providing cochlear implant therapy to the prelingually deaf children as early as possible .Methods A total of 55 pediatric relingually deaf cases were included in this study .According to their implantation time and application duration ,they were divided into 2 groups :1 to 3 years old group (32 cases) ,and >3 to 5 years old group (23 cases) respectively .The hearing ,lan-guage and learning abilities on 1 ,3 ,6 ,12 ,18 ,24 months after cochlear implantation were evaluated ,using statisti-cal method to record CAP and SIR scores .Results The rehabilitation effects ,the average ages ,CAP ,speech rec-ognition rates and SIR were increased two years afterwards .The effects of younger age group were more noticeable than that in the older group .The differences between the two groups were statistically significant (P<0 .05) .Av-erage speech recognition rates ,average speech rehabilitation effects in each postoperative period of younger age group were better than those of in older age group ,showing significant differences (P<0 .05);CAPs in the younger age group on 1 ,3 and 12 months after CI surgery were significantly higher than those of in the older group (P value were 0 .001 ,0 .001 and 0 .002 ,respectively) .SIR in the younger age group at the time of 1 ,3 ,12 ,24 months were significantly higher than those of in the older group(P values were 0 .00 ,0 .00 ,0 .00 and 0 .024 ,respectively) . Conclusion Implanted age and recovery time are the key factors that influence the effects of postoperative rehabilita-tion .The younger when the children get cochlear implantation and the longer the recovery time takes during two years after cochlear implantation ,the better the results are .The standardization of domestic assessment for the re-covery effects and the international evaluation method have a certain degree of equivalence .

Objective To study the expression profile of microRNAs in the lung adenocarcinoma tissue and its expression in CD133+ lung cancer cells .Methods The specimens of adenocarcinoma tissue and pericarcinomatous tissue were collected for con-ducting the microRNA expression chip analysis .The flow cytometry(FCM ) was adopted to sort and culture CD133+ A549 cells in serum-free medium for 2 weeks .The positive rate of CD133+CD44+ in those cells was assayed .The expression of miRNA-892b and miRNA-4686 in CD133+ A549 cells were detected by real-time qPCR .Results Compared with the pericarcinomatous tissue ,27 miRNAs expressions in the lung adenocarcinoma tissue were up-regulated(P<0 .05) and 6 miRNAs expressions were down-regula-ted(P<0 .05) .The positive rate of cancer stem like cells CD 133+CD44+ was 75 .0% .The expression of miRNA-892b and miRNA-4686 in CD133+ A549 cells was higher than A549 cells(P<0 .01) .Conclusion Abnormal expression profile of miRNAs in lung ad-enocarcinoma tissue ,and miRNA-892b and miRNA-4686 may play a certain role to maintain the biological characteristics of CD133+ lung cancer cells .

Adenocarcinoma ; metabolism ; pathology ; Adenocarcinoma, Follicular ; metabolism ; pathology ; surgery ; Bronchial Neoplasms ; metabolism ; secondary ; surgery ; Carcinoid Tumor ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Middle Aged ; Thyroglobulin ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology ; surgery ; Transcription Factors

Objective To investigate the chemical properties of superficial cardiac neurons. Methods By means of immunohistochemical ABC technique,the study was performed concerning the distribution of calcitonin gene-related peptide(CGRP) and substance P(SP) in the canine main cardiac superficial plexus. Results CGRP-immunoreactive(IR) neurons were found in every plexus,but SP-IR neurons could be observed only in dorsal atria plexus(DAP),inter atria plexus(IAP) and aorta-pulmonary plexus(A-PP).The shape and size of CGRP-IR and SP-IR neurons were similar.The comparative study on atria and ventricles indicated that CGRP-IR and SP-IR neurons in atria were more than those in ventricles.Numerous CGRP-IR,SP-IR nerve fibers could be observed in each fat pats and intermyocardiocytes.These nerve fibers were usually situated near blood vessels,or were attached to vessel wall.Somewhat CGRP-IR and SP-IR nerve fibers were connected with myocardiocytes in some regions.Conclusion The results indicated that actually existed two peptides in canine cardiac superficial plexus.These implied that the regulations of the two kinds of peptidergic neurons to atria and ventricle were different.The two kinds of neurons mentioned above were likely to perform different or similar functions in canine heart.CGRP and SP could possibly modulate the activities of myocardiocytes and vessels of heart directly.

The objective of this study was to characterize the genome structure of duck hepatitis B virus (DHBV) isolated from Hubei brown ducks. The natural carrier rate of DHBV in adult ducks from Hubei area was investigated and the DHBV DNA-positive serum screened out. The complete genome of a DHBV strain was amplified by polymerase chain reaction (PCR) and cloned into T vector and sequenced. The results showed that the carrier rate of DHBV in Hubei brown ducks was 10 %. This strain (GenBank accession number DQ276978) had a genome of 3024 nucleotides with three overlapping open reading frames encoding the surface, core and polymerase proteins respectively. Comparison of the strain with 17 DHBV strains registered in GenBank revealed a homology from 89.3 % to 93.5 % at the nucleotide level. The sequences of the structural and functional domains of these proteins were highly conserved. The strain was found to share more signature amino acids in the polymerase genes with the "Chinese" DHBV strains than those of the "Western" country strains. This finding was also corroborated by a phylogenetic tree analysis. Therefore, the DQ276978 might belong to a subtype of the Chinese DHBV strains.

Objective To explore the role of CD4+ CDhigh25 regulatory T cells in the pathogenesis of chronic abacterial prostatitis/chronic pelvic pain syndrome (CAP/CPPS).Methods The percentage of CD4+ CD+25 and CD4+ CDhigh25 regulatory T cells was detected by flow cytometry from 45 CAP/CPPS pa-tients and 18 normal controls.The levels of interleukin-6(IL-6),IL-10,tumor necrosis factor-α(TNF-α) and transforming growth factor-β1 (TGF-β1) in serum and seminal plasma were measured by ELISA in the same cohort.Results There was no significant difference in the percentage of peripheral blood CD4+CD+25 and CD4+Cdhigh+ cells between CAP/CPPS patients and normal control (P>0.05).The ser-CD4+CD+25 and CD4+Cdhigh+ cells between CAP/CPPS patients and normal control (P>0.05>.The ser-um levels of TGF-β1 in patients with CAP/CPPS were markedly lower than those in controls (P<0.05),serum TNF-α and seminal plasma IL-6,TGF-β1 and TNF-α in CAP/CPPS patients were markedly higher than those in controls (P<0.05).There was a positive correlation between the IL-6 and the NIH-CPSI.There was also a positive correlation between the IL-10 and the pain index.In ad-dition,the percentage of peripheral blood CD+4CDhigh25 cells was positively correlated with serum TGF-β1.But the percentage of CD+4CDhigh25 cells had no correlation with ages,duration of CAP/CPPS pa-tients,NIH-CPSI and the other cytokines.Conclusions The defective function of peripheral blood CD+4CD+25 regulatory T cells may be related with the pathogenesis of CAP/CPPS.The cytokines may also play an important role in the process of pathogenesis of CAP/CPPS.

Objective: To screen and identify serum amyloid A (SAA) in patients with prostate cancer with mass spectrum technique. Methods: SELDI technology was used to detect the changes in protein expression. SAA was screened and separated and then identified by peptide mass fingerprint (PMF) based on matrix-as-sisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. Results: The analysis showed that a mass cluster in the ranges of 11.1-11.9KD in M/Z value in the serum of prostate cancer patients was much higher than that in the control group. Meanwhile, this protein peak was closely correlated with clinical stages of prostate cancer. The level of the protein peak was increased as the illness got serious. Through MALDI technology combined with HPLC, the mass cluster in the range of 11.1-11.9KD in M/Z value on the chip was identified as SAA. And it was also verified through ELISA method. Conclusion: Mass spectrum technology is an effective method to detect the biological markers in prostate cancer patients. This method is convenient, highly sensitive and with good reproducibility. The SAA can be used as a marker in the diagnosis of prostate cancer. These indices are also meaningful in screening and identifying signal proteins from the serum of prostate cancer patients.