Fibroma, Desmoplastic ; pathology ; Humans ; Male ; Middle Aged ; Nose Neoplasms ; pathology

Objective:To prepare monoclonal antibodies specifically against an immunogenic fragment in ectodomain of prostate-specific membrane antigen ( PSMA ).Methods: An polypeptide immunogenic fragment in the ectodomain of PSMA was predicted by biological information technology,and then it was expressed prokaryotically.BALB/c mice were immunized with the prokar-ytically expressed recombinant polypeptide antigen,to prepare the monoclonal antibodies specifically against an immunogenic fragment in ectodomain of PSMA by hybridoma technology,purification of monoclonal antibody by affinity chromatography,characterization of the monoclonal antibodies by Western blot.The radioimmunoimaging in prostate cancer model was performed by using the labeled McAb.Results:Throught the software analysis,we got the antigen fragment in the ectodomain of PSMA containing 310aa sequences higher specificity, artificially synthesized gene sequence of the region, and constructed a prokaryotic expression vector pET-32a-r-ectodomain-PSMA,by prokaryotic expression we obtained the 50 kD target antigen,after hybridization,the three positive hybridoma cell lines (5E6,4A5 and 4D7) were selected by ELISA using target antigen,the isotypes of 5E6 and 4A5 were IgG2a,the isotypes of 4D7 were IgG1,the titer of three monoclonal antibodies was above 1∶256 000.Western blot results showed that the prepared monoclonal anti-bodies could binding specifically to the antigen in the ectodomain of PSMA.Radioimmunoimaging in prostate cancer animal model results further confirm that the prepared monoclonal antibodies could combinate with the antigen in the ectodomain of PSMA in the animal body, and make the tumor imaging.Conclusion: The prepared monoclonal antibodies can specifically recognizes the PSMA antigen,which laid the foundation for the immunodiagnosis and immunotherapy of prostate cancer.

Objective To evaluating the effectiveness of comprehensive measures to control the plague in epidemic areas in Longlin county Guangxi from 2001 to 2010.Methods Original epidemic places was deratised,indicative animals were investigated,and epidemic clues were searched.Cage trapping method was used to capture rat and rat body fleas were collected in the plague epidemic areas.The flea-carrying rates and flea index of rodents were calculated based on the number of fleas collected from caged rodents.The animals were then subjected to etiological and serological tests to determine the plague infection rate.Results A total of 1008 rats were captured and 571 fleas were collected from 2001 to 2010,of which Rattus Flavipestus accounted tor 81.65％(823/1008) and Xenopsylla Cheopis for 64.10％(366/571).The annual average rodents infected with flea and the index of flea were 23.02％ (177/769) and 0.74,respectively.The annual average density of rodents decreased from 3.99％ (859/21 508,before deratised) to 0.96％ (149/15 600,after deratised).The deratization rate was 75.94％.Conclusion The risk of a plague epidemic in Longlin county is reduced after continued comprehensive measures be taken to deal with the disease.

Objective To explore a better method for treatment atrophic rhinitis.Methods 56 patients with atrophic rhinitis(96 lateral)were treated by nasal submucou pediculated bone-suberiosteal muscle flap extracted from anterior wall of sinus maxillaries.Results All patients were followed 2 to 10 years,total effective rate was 100 %, with 49 cases(87.5 %)showing prominent effect.Conclusion The grafted flap cannot be assimilated,felled off and necrosis,because the flap has rich blood supply.This methods has obvious short-term effective and stable long-term effective.No complications were found.

Background Retinitis pigmentosa (RP)is a common hereditary blinding eye disease in ophthalmology.Current researches documented that RP may have the common pathophysiologic basis to Alzheimer disease and chronic neurodegenerative disease.Understanding this mechanism will offer a new therapeutic target for RP.Objective The purpose of the present study was to investigate the roles of cyclin-dependent kinase 5 (Cdk5)/P25 activation in the apoptosis of retinal neural cells of RCS rats.Methods Eighteen SPF RCS rats and 18 RCS-rdy+ rats were randomized into 17-,25-and 35-day groups respectively and 6 rats for each.The rats were sacrificed at corresponding time points and retinal hemogenete was prepared.Expressions of CdkS,P35,P25 and tau phosphorylation in the retinas were detected by Western blot,and the kinase activity of Cdk5/P25 was analyzed by quantitative colorimetric assay.Results The expressing level of P35 protein(A340) in the retinas of 17-day-old RCS rats was near that of 17-day-old RCS-rdy+ rats(t =0.52,P＞0.05).In 25-and 35-day-old RCS rats,the expressing levels of P35 protein were 2.20±0.48 and 1.23±0.14,which were higher than those of RCS-rdy+ rats(1.43±0.13 and 0.93±0.10),showing significant differences between them(t =3.78,4.28,P＜0.05).The expression of P25 was undetectable at postnatal 17 days in RCS rats and RCS-rdy+ rats,but it showed significantly higher in RCS rats(0.300±0.003 and 0.230±0.004) than that in RCS-rdy+ rats(0.040±0.004 and 0.070±0.004) at postnatal 25 days and 35 days(t=121.81,77.51,P＜0.01).No significant difference was found in the expression of Cdk5 in RCS rats and RCS-rdy+ rats at different ages (t =-0.60,0.19,1.62,P＞ 0.05).The kinase activity of Cdk5/P25 did not show significantly different between RCS and RCS-rdy+ rats at postnatal 17 days(t =0.19,P＞0.05),but significantly higher kinase activity of Cdk5/P25 was seen in RCS rats (0.0058 ±0.0005 and 0.0056±0.0004) than that in RCS-rdy+ rats(0.0038±0.0003 and 0.0032 ±0.0007) at postnatal 25 days and 35 days (t =8.07,5.97,P＜ 0.01).No expression of tau phosphorylation was detected in RCS rats at postnatal 17 days,but significantly higher tau phosphorylation level was seen in RCS rats at postnatal 25 days and 35 days(1.80±0.22 and 1.23±0.17),which were significant different in comparison with RCS-rdy+ rats at postnatal 25 days and 35 days(1.60 ±0.20 and 1.04 ±0.12)(t=4.71,3.17,P＜0.05).Conclusions The Cdk5/P25 kinase activity shows a consistent trend with theexpressions of P25 and tau phosphorylation in the RCS rats,indicating that the upregulation of P25 induces the enhance of enzyme activity of Cdk5,which phosphorylate its substrates to result in more apoptosis of retinal neural cells.

OBJECTIVETo evaluate serum-vascular endothelial growth factor (S-VEGF) in the differentiation of solitary pulmonary nodule (SPN).

METHODSSerum level of VEGF of 68 patients with SPN was measured by ELISA kit, and compared with the control group of 20 normal subjects. The nodules were diagnosed by operation and pathology.

RESULTSThe median level of S-VEGF was 42.5 (range from 10 to 170) pg/ml in the control, 44 (range from 18 to 360) pg/ml in benign nodule group and 75 (range from 18 to 890) pg/ml in lung cancer group, with significant difference observed between the nodule group and control (P < 0.01), and between the lung cancer group and the benign nodule group (P < 0.05), but not between the benign nodule group and the control. In addition, when S-VEGF in different pathologic types of the limited number of lung cancer patients were compared, no significant difference was observed.

CONCLUSIONS-VEGF is valuable in the differential diagnosis of solitary pulmonary nodule. An elevated S-VEGF level >or= 100 pg/ml in patients with SPN may strongly speak for a malignant nodule. Operation is suggested.

Adult ; Aged ; Diagnosis, Differential ; Female ; Humans ; Lung Neoplasms ; blood ; diagnosis ; Male ; Middle Aged ; Solitary Pulmonary Nodule ; blood ; blood supply ; diagnosis ; Vascular Endothelial Growth Factor A ; blood

Background Retinitis pigmentosa (RP) is a monogenic inheritance and blinding disease of fundus oculi.There is not an effective therapeutic method now.Objective This work was to identify the mutations of RP1 gene in Chinese RP patients in Ningxia area and to explore the potential interactions in the pathogenesis of RP.Methods The periphery blood of 3-5 ml was collected from 110 individuals with RP(35 ADRP and 75SRP)and 100 normal controls in Ningxia area.Polymerase chain reaction (PCR) and direct DNA sequencing were used to screening the sequence alterations in the entire coding region and splice sites of RP1 gene.Multivariate analysis and two web-based programs( PolyPhen and SIFT) were used to analyze the results.Results Eleven mutation locus were detected in the exon 4 of RP1 gene including two novel sequence variants:p.Lys1152Lys without a higher mutation rate in comparison with normal control group(x2 =9.12 P＜0.01 ),but c.* 247A＞C with a higher mutation rate in comparison with normal control group(x2 =12.77,P＜0.01 ) and c.* 247A＞C mutation was thought to be correlated with RP( r=1.11,P＜0.05 ).The other ten mutation locus were reported as single nucleotide polymorphisms (SNP).The mutation rate of p.Gln1725Gln was found to be higher in the RP patients than the normal controls (x2 =42.09,P＜0.01 ),but no the significant correlation was seen between the pathogenesis of RP and mutation of p.Gln1725Gln(r=1.74,P＞0.05).p.Lys1152Lys mutation was found in only 1 patient.Three SNPs( p.Arg872His,Ala1670Thr,Ser1691Pro) were always occurred in the same 83 RP patient and the relevance ratio was higher than controls ( P＜0.01 ).The age of night blindness on patients with concurrent three mutations was (30.54± 13.68 ) years,and the best corrected visual acuity (BCVA) was 0.50 ± 0.38.The age of night blindness on patients without concurrent three mutations was(21.06± 16.24) years,and the BCVA was 0.40 ±0.33 and were higher than controls ( t =2.11,P ＜ 0.05 ).Conclusions In this study,the prevalence of RP1 mutations among the RP patients in Ningxia population was lower than other populations (＜ 1％ ).The alliance of SNPs (p.Arg872His、p.Ala1670Thr、p.Ser1691Pro) may play a protective role on RP patients and reduce the frequency of mutatiaon in RP1 gene.

Objective To analyze the outcome of surveillance results on plague and to provide the evidences for the policy making in Longlin county Guangxi. Methods The epidemic data and the surveillance results of plague were analyzed and assessed with epidemiology methods in Longlin county Guangxi from 2000 to 2009, and the density of rodents, the rodents infected with flea, flea index and other indicators were calculated. Regional composition of the rats and fleas were analyzed. Results A totally of 4829 rats were captured and 4737 fleas were collected in the past 10 years, Rattus Flavipestus(81.92%,3956/4829) and Xenopsylla Cheopis (79.04%,3744/4737) were dominant species. The annual average density of rodents, the rodents infected with flea, index of flea were 3.30%(4829/146 206), 27.99%(1351/4827) and 0.98(4737/4827), respectively. A totally of 4792 rats were examined and 10 strains Yersinia Pestis were isolated. Indirect hemorrhagic assessed(IHA) was used to test the F1 antibody against plague in the blood serum of the rats and indicator animals, and 3 positive rats and 24 positive animals were found, respectively. Twenty seven natural villages in 3 towns had been involved in the plague. Conclusions The plague foci exists in Longlin county of Guangxi province. The plague foci in the areas have the same feature with the plague foci of Rattus Flavipectus. There is a potential risk for plague in this region, we should improve the quality of surveillance, increase indicator animals of the plague, and try to apply new surveillance method.

OBJECTIVETumor necrosis factor alpha-stimulated gene-6 (TSG-6 gene) differentially expressed in adipose tissue of obese and normal human subjects or rats. To explore the relationship between the differential expression of TSG-6 and adipocyte differentiation, adipogenesis and obesity, the present study aimed to investigate the changes of TSG-6 gene expression during 3T3-L1 preadipocyte differentiation and to analyze the regulative role of TNF-alpha on TSG-6 gene expression in matured 3T3-L1 adipocytes.

METHODS3T3-L1 preadipocytes were cultured in vitro and differentiated into the matured adipocytes. TNF-alpha in different concentrations (0.1 ng/ml, 1.0 ng/ml, 10.0 ng/ml) was added into the culture medium of fully differentiated adipocytes (day 10) for various times (0.5 h, 2 h, 6 h, 12 h, 24 h). Total RNA from these adipocytes was extracted and the levels of TSG-6 gene mRNA expression were evaluated by RT-PCR.

RESULTS(1) In preadipocytes, the level of TSG-6 gene mRNA expression remained low. In the presence of dexamethasone (Dex), MIX and insulin, with the 3T3-L1 preadipocytes being differentiated into the matured adipocytes, the level of TSG-6 gene mRNA expression was upregulated and reached the higher level in fully differentiated adipocytes. There is a significant difference between any two detected phases in the levels of TSG-6 gene mRNA expression (P < 0.05), except that the levels of TSG-6 gene mRNA expression did not increase obviously on day 0 to day 2, day 3 to day 5, day 4 to day 6 and day 7 to day 10 (P > 0.05). (2) Treatment of day 10 3T3-L1 adipocytes with TNF-alpha of different concentrations (0.1 ng/ml, 1.0 ng/ml, 10.0 ng/ml) resulted in a significant decrease in the level of TSG-6 gene mRNA expression. The inhibition effect of TNF-alpha on TSG-6 gene mRNA expression generally tended to be reinforced with the increasing concentrations of TNF-alpha and the elongation of time course, except for the period of 6 - 24 h after the stimulation of 10.0 ng/ml TNF-alpha. When 0.1 ng/ml TNF-alpha was applied, the level of TSG-6 gene expression decreased by 33.73% at 6 h, 97.39% at 12 h. While 1.0 ng/ml TNF-alpha was used, the level of TSG-6 gene expression decreased by 78.68% at 6 h, which remained until 24 h. At a concentration of TNF-alpha up to 10.0 ng/ml, the level of TSG-6 gene expression decreased by 96.27% at 2 h. TSG-6 gene expression was almost fully inhibited.

CONCLUSION(1) TSG-6 gene may be involved in adipocyte differentiation and adipogenesis. (2) TNF-alpha can downregulate the mRNA expression of TSG-6 gene in matured adipocytes. The inhibitory effect of TNF-alpha on TSG-6 gene expression is generally dose-correlated.

3T3-L1 Cells ; Adipocytes ; cytology ; drug effects ; metabolism ; Animals ; Cell Adhesion Molecules ; genetics ; Cell Differentiation ; drug effects ; genetics ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Mice ; RNA, Messenger ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors ; Tumor Necrosis Factor-alpha ; pharmacology

A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood. The technology utilizes special primers and Taqman MGB fluorescence probe to measure amplification products from the gag-pro-pol polyprotein gene of HTLV-I. HTLV-I copy number was normalized to the amount of cellular DNA by quantitation of the beta-actin gene, The amplification system was sensitive to detect 5 copy/microL. The standard curve had a good linearity when the quantity for the gene was between 10(3) and 10(7) copy/microL (R2 = 0.999). Good reproducibility was observed in each intra- and inter-assay. We also measured proviral load in peripheral blood in 12 HTLV-I seropositive former blood donors. Proviral load for HTLV-I infected donors ranged from 0.015 to 12.819 copy/cell in WBC with the mean of 3.116 copy/cell.
Gene Products, gag ; genetics ; Gene Products, pol ; genetics ; Human T-lymphotropic virus 1 ; genetics ; isolation & purification ; Humans ; Molecular Probes ; Polymerase Chain Reaction ; methods ; Viral Proteins ; genetics