Objective To explore whether the neural stem cells(NSCs) can act directly as a gene target cell which can be infected by the recombinant retrovirus and express the products of exogenous genes after infection. Methods The NSCs were cultured with supernatant containing the recombinant retroviruses with the genes of NGF or GDNF for two days.After screened with G418,the infected NSC were expanded at the present of bFGF in culture.The PC12 cells and the neurons of ventral midbrain of rat were cultured by the medium from the infected NSC,which were called as GDNF\|containing conditioned medium NGF or GDNF\|containing conditioned medium the morphological changes of the dopamine neurons of the ventral midbrain and expression of exogenous genes of the infected NSCs were detected by immunohistochemistry staining. Results It was estimated that about fifty percent of NSCs via retrovirus\|mediated NGF or GDNF gene transduction were G418\|resistant.These infected NSCs began to differentiate.Long and radical processes reached out from the sphere of proliferation and the cells migrated towards outside along the processes.The NSC infected with gene of NGF showed an astroid\|shape with larger body and processes.The NSC infected with gene of GDNF showed a shuttle\|shape with a smaller body and long processes.The PC12 cells increased in the NGF\|containing conditioned medium and stretched out long neurites.The dopamine neuron of the ventral midbrain which were immunoreactive for TH also showed a larger body and longer processes in the GDNF\|containing conditioned medium.Most of G418\|resistant NSCs were immunoreactive for NGF or GDNF. Conclusion NSC can act directly as a gene target cell which not only be infected by the recombinant retrovirus,but also express and secrete the products of exogenous genes.

Through genetic recombination technique, the rat glial cell line-derived neurotrophic factor (rGDNF) cDNA was in-serted into polylinker site of retroviral vector pLXSN, to generate a recombinant plasmid pLXSN-GDNF as transfer vector. Therecombinant plasmid was verified with restriction analysis, PCR, dot blot hybridization and Southern blot hybridization. The re-sults showed that GDNF cDNA was cloned correctly into retroviral vector pLXSN, recombinant retroviral vector was construct-ed. It is concluded that the eukaryotic cell expression vector was constructed successfully for gene therapy of Parkinson's,Alzheimer's and other central nervous system diseases.

In order to investigate effect of basic fibroblast growth factor(bFGF) on the prolifiration and differentiation of theneural progenitor of embryonic hippocampus of rats in vitro, 25 ng/ml bFGF was employed into the serum-free medium of cultureof hippocampal neural cells of embryonic day 18 rats in the present study. The effect of bFGF on the viability of cells culturedwas detected by MTT colorimetric method and the effects of bFGF on proliferation and differentiation of hippocampal neural pro-genitors were analyzed qualitatively and quantitatively by means of immunochemistry, for the nestin, neurofilament, galactocere-broside and glial acidic fibrillary protein. The results showed that the OD value of experimental group was higher than that ofcontrol group by 1.5 and 1.8 times at 4 d and 8 d respectively. The quantitative analysis of each kinds of cells indicated that thenumber of neural progenitors, neurons and oligodendrocytes in experimental group were increased about 2 times as many as thatin control group, but no differences of astrocytes between the two groups at 4 d. However, the number of four kinds of cells aug-mented about 1.7 times at 8 d. The results of this study suggest that bFGF can not only promote the survival, proliferation, butalso facilitate the differentiation of neural progenitor of hippocampus to neurons and glial cells. To obtain more many purifiedneural progenitors in vitro, the embryonic day 18 is not an appropriate age. It is better to get younger embryo brain to culture and to add enough bFGF.

OBJECTIVE An understanding of the complex anatomy of the anterior skull base is crucial for the surgeon doing endoscopic surgery. The anatomic data of the olfactory sulcus and adjacent structures in Chinese patients were defined using 3-dimensional reconstruction images. The surgeon is encouraged to develop a detailed pre-operative surgical plan by utilizing these dynamic anatomical observations to avoid intracranial injury. METHODS The paranasal sinus CT scanning images of 100 adults were reconstructed for observation using EBW2.0 software and multiplanar reformation. All data obtained were in the coronal plane from the anterior point of the olfactory sulcus. The cribriform plate depth as compared to the ethmoid roof and adjacent structures, was measured bilaterally. Data obtained on adjacent structures include the vertical height of the lateral lamella of olfactory sulcus, the horizontal distance between the cribriform plate and the orbital lamella, the length of the middle turbinate, the height of the orbit, and the vertical distance between the cribriform plate and the nasal floor. RESULTS The olfactory sulcus was classified into three types: platform type (60 %), sloping type (17 %) and mixed type(23 %), as distinguished from Keros classification. In this study the vertical height of the lateral lamella of olfactory sulcus was (5.03 ± 0.17) mm (R) and (5.39 ± 0.19) mm (L) in platform type, and (2.79 ± 0.49) mm (R) and (4.72 ± 0.49) mm (L) in the mixed type. There were statistically significant differences between the right side and the left side in these two types (P＜0.01). The horizontal distance between the cribriform plate and the orbital lamella on the same side was significantly different between the platform type and the mixed type of olfactory sulcus. A similar result was observed for the vertical distance between the cribriform plate and the nasal floor. Gender differences exist in the horizontal distance between the cribriform plate and orbital lamella on the same side and the vertical distance between the cribriform plate and the nasal floor. CONCLUSION Different types of olfactory sulcus have distinct characteristics, hence care which must be taken into account when doing endoscopic surgery.

Objective To evaluate the serum level of antimicrobial peptide human cationic antimicrobial protein 18 ( hCAP18 ) in non-small cell lung cancer ( NSCLC ) patients and its auxiliary diagnosis and prognosis value.Methods Case-control study was used.The serum level of hCAP18 was measured by enzyme linked immunosorbent assay ( ELISA) in 50 cases with NSCLC patients of department of thoracic surgery and 50 cases healthy people of department of physical examination from January 2011 to January 2012 in Tongji Hospital of Tongji University.The concentrations of hCAP18 in serum of NSCLC patients before and after surgery were analyzed.The sensitivity and specificity of serum hCAP18 for the diagnosis of NSCLC were evaluated using the receiver operating characteristic ( ROC ) curves.Data was analyzed by using the t-test and Log-rank test.Results Serum hCAP18 concentration in NSCLC patients (6 733 ±771.8) μg/L was significantly higher than in healthy controls (253 ±6.9) μg/L (t=8.396, P<0.05) .However, the concentration of hCAP18 showed no significant difference between squamous cell carcinoma and adenocarcinoma[(6 300.0 ±1 221.0) μg/L and (7 074.0 ±1 005.0) μg/L, respectively;t=0.494 2, P <0.05 ] .hCAP18 levels had significantly decreased in serum of NSCLC patients after 30 d surgery compared to preoperative results[from (6 733.0 ±771.8) μg/L to (433.6 ±38.2)μg/L;t=8.512, P<0.05].ROC analysis of serum hCAP18 yielded an AUC (Area under the ROC curve) of 0.931 ( 95% CI =0.884 -0.978 ) with 95% sensitivity and 96.3% specificity, which was higher than the CYFRA21-1[0.873 (95%CI=0.758-0.917)].The relapse rate of NSCLC patients with serum hCAP18≤390.0 μg/L was 12.5%(4/32), while 44.4%(8/18) in NSCLC patients with serum hCAP18>390.0μg/L (χ2 =22.64,P<0.05).Conclusions Detection of serum hCAP18 shows a good sensitivity and specificity for the auxiliary diagnosis of NSCLC. It is possible to be a potential detection index for noninvasive diagnosis and monitoring progression of lung cancer.

Objective To establish a TaqMan-MGB fluorescent probe characterized real-time polymerase chain reaction ( qPCR) method for detecting retinoic acid induced genes G ( RIG-G) in human acute promyelocytic leukemia ( M3 ) .Analyze RIG-G expression levels in peripheral blood of both normal persons and M3 patients and explore its diagnosis value for M 3.Methods Methodology establishment study.A detection method and standard curve of TaqMan-MGB real-time PCR were established after designing specific primers and TaqMan-MGB fluorescence probe of human RIG-G gene and using reverse transcription complementary DNA ( cDNA) as a template.The performance of this method was evaluated in specificity, accuracy, precision, analytical sensitivity and interference substances . Twenty clinical specimens with M3 were quantified RIG-G expression so as to evaluate the correlation between peripheral blood and bone marrow samples .Meanwhile , the results of RIG-G expression in peripheral blood of 40 normal specimens and 20 patients with M3 were analyzed by t-test.And receiver-operating characteristic curve ( ROC ) was used to analyze the detection efficiency of M 3.Results There was a good linear relationship between log value of RIG-G standard substance and threshold cycle number ( Ct ) ( standard curve equation:Y=-3.539X+42.952,R2 =0.999).New method was used to detect standard substance . The deviation between observed and expected values was <5% (r=0.999).Three concentration samples (107 ,104 ,101 copies/μl) were selected for precision test.Intra-assay coefficients of variation were 1.38%, 2.31% and 1.38%, respectively , and intre-assay coefficients of variation were 0.71%, 1.17% and 5.07%, separately.All were less than 10%.The sensitivity of this method was 101 copies/μl.There was a good correlation of RIG-G results between peripheral blood and bone marrow in M 3 patients(r=0.996, b=0.973).But there was no significant difference between this two group results (t=0.099, P>0.05). However , there was obvious difference of RIG-G value in peripheral blood between control group and M 3 patient group (U=18,P<0.001), 3.62 ×104(1.61 ×104 -4.90 ×104)copies/μl for controls and 7.10 ×102 (5.43 ×102 -2.21 ×103 ) copies/μl for M3 patients, respectively.Conclusions Successfully establishe a TaqMan-MGB real-time PCR method for detecting RIG-G gene in peripheral blood.The accuracy, precision, sensitivity and specificity are good .It could provide necessary help in early diagnosis and monitor treatment of clinical M3 patients.

Objective To investigate the survival,differentiation and gene expression of the neural stem cells(NSCs) modified by the gene of NGF or GDNF in the brain of AD rat model after transplantation. Methods The NGF or GDNF genetically modified NSCs labled with BrdU were implanted into the lateral cerebral ventricle of the AD rat model.The rats were killed three weeks after transplantation.The brains were cut and the sections were processed for single or double immunochemistry staining with antibodies against BrdU,Nestin,GFAP,NF,NGF and GDNF. Results BrdU\|positive cells were found in the lateral cerebral ventricle.Some of them migrated into the parenchyma and located in the wound,fibria\|fonix,hippocampus,corpus callosum,septum,subventricle zone and beside the blood vessels.Cells of doubled labeling with anti\|BrdU and GFAP were more often seen in the cortex,whereas more cells with anti\|BrdU and NF in the hippocampus,and both of them in the ventricle.Doubled labeling cells against BrdU and NGF,and against BrdU and GDNF were found in the ventricle and parenchyma.Conclusion\ The NGF or GDNF genetically modified NSCs can not only survive well,but also differentiate into neuron and astrocyte,and express the exogenous genes of NGF and GDNF in the host brain

Objective To investigate the effect of a single or combined transplantation of the neural stem cells(NSCs) modified with gene of NGF or GDNF on the cholinergic neurons of basal forebrain of AD model rat. Methods The NSCs modified with gene of NGF or GDNF were implanted in single or combined into the lateral cerebral ventricle of the rats after fibria\|fornix transection.The rats were killed three weeks after transplantation and the brain sections concluding basal forebrain were cut coronally on a freezing microtome and were processed by immunohistochemistry staining with antibodies against ChAT.The numbers of ChAT positive neurons of medial septum(MS) and vertical diagonal band(VDB) were analyzied statistically with one way of Student\|Newman Kaels. Results In MS,the percentages of ChAT positive neurons at the lesion side to the intact side in NGF group was 81% which was significantly higher than that in the lesion group(34%),NSC group(36%) and GDNF group(50%), P 0\^05). Conclusion\ The injury cholinergic neurons can be protected in different extent after a single or combined transplantation of the neural stem cells modified with gene of NGF or GDNF.Among these three groups,greater protection was found in NGF group and NGF+GDNF group,and lesser protection in GDNF group.\;[

Objective To investigate the memory amelioration of the AD model rat after a single or combined transplantation of the neural stem cells(NSCs) modified with gene of NGF or GDNF. Methods The AD model rat was made by cutting unilaterally the fibria\|fornix of male SD rat.Eight to ten days after surgery,the genetically modified and unmodified NSCs were implanted into the lateral cerebral ventricle of the rats.Two weeks after transplantation,the amelioration of memory impairment of the rats were detected by Morris water maze. Results The average escape latencies of the last three blocks in NGF and NGF+GDNF groups were lesser,and the percentages of swim distance in the platform quadrant were greater than that in NSC group( P

Background and purpose:Retinoic acid-induced gene G (RIG-G) is a tumor suppressor gene which is cloned by NB4 cell line from a acute promyelocytic leukemia cell. This study aimed to investigate the effect ofRIG-G in lung cancer cells A549 by constructing a lentiviral vector expressing RIG-G under doxycycline (DOX) regulation.Methods:RIG-G gene ampliifcation was performed by quantitative real-time PCR (qRT-PCR). pLenti6/TO/V5-GIM-RIG-G lentiviral vector withGFP was built by LR recombination system. The concentration of pLenti6/TO/V5-GIM-RIG-G lentiviral vector andTet-on lentiviral vector were measured by virus titer method. After infecting A549 cells, stably transfected lines were selected via limiting dilution analysis.RIG-G gene expression was examined by immunolfuorescence staining and Western blot assay. Cellular proliferation was determined by CCK-8 assay.Results:The concentrations of pLenti6/TO/V5-GIM-RIG-G lentiviral vector andTet-on lentiviral vector were 1.0×108TU/mL and 4×109 VP/mL, respectively. RIG-G was expressed in lentivirus infected A549 cells after adding DOX, and the amount of cells withGFP could be observed by lfuorescence microscopy.After the expression of RIG-G protein, the prolif-eration activity of A594 cell was signiifcantly inhibited compared to the control group (1.168±0.107vs 2.099±0.162, P<0.05).Conclusion:The regulated expression ofRIG-G gene was established in A549 lung cancer cell line. The RIG-G protein has potential abilities to inhibit the proliferation of lung cancer cell A549.