OBJECTIVE To evaluate the decontamination capability of hydrogel polymer coated ZnO nanoparticles (ZnO NP-gel) against soman. METHODS ZnO NP was synthetized using chemical precipitation method and modified with 4-pentenoic acid,and then polymerized with comonomers to obtain ZnO NP-gel. The transmission electron microscope (TEM), scanning electron microscope (SEM) and particle size instrument were used to observe the internal structure,micromorphology,particle size and zeta potential of these materials. An infrared spectroscope (IR) was used to analyze their chemical bond structure,while X-ray diffraction (XRD) was used to analyze the diffraction pattern.The content of soman was determined by benzidine chromogenic reaction. ZnO NP(1 g·L-1), ZnO NP-gel (1 g·L-1) and distilled water were mixed with soman(52.2 mg·L-1),stood for 30 min,and then filtered before filtrate was subcutaneously injected into mice (40 μL·g-1) to observe the symptoms of poisoning and death. RESULTS SEM and TEM showed that ZnO NP-gel had a block structure, the zeta potential of which was (-7.89 ± 0.04) mV. The results of IR indlicated that ZnO NP-gel had stronger absorption peaks at 754 and 618 cm-1, and XRD revealed that these materials had a sharp peak at 2θ=8.06738°. The decontamination efficiency of ZnO NP-gel was higher than that of ZnO NP group at the same concen?tration (n=3, P<0.05), and the time for decontamination of 50% soman was shortened by four times. The mice were injected subcutaneously with the soman solution treated with ZnO NP-gel, which caused no convulsion or death. CONCLUSION ZnO NP-gel can perform the double function of fast adsorption and catalysis of soman,and the decontamination ability of which could be improved through polymer modification.

OBJECTIVETo investigate the effect of bufalin on nucleus-mitochondria localization of human telomerase reverse transcriptase(hTERT) by exploring its effect on proliferation and apoptosis in human esophageal squamous carcinoma EC9706 cells.

METHODSEC9706 cells were treated with bufalin at various concentrations, and then the cell growth inhibition of EC9706 cells was examined by CCK-8 assay and the 50% inhibitory concentration (IC(50)) was calculated.Cell cycle analysis was performed by flow cytometry with PI staining, and nucleus morphology of apoptosis were observed by fluorescence microscopy with Hoechst 33342 staining. The apoptotic index was measured by flow cytometry with Annexin V-FITC/PI double staining. hTERT subcellular localization and protein expression were determined by Western blotting and multiple immunofluorescence labling combined with laser confocal scanning microscopy.

RESULTSThe proliferation of EC 9706 cells was significantly inhibited by bufalin along with the increase of processing time and concentrations (p<0.01). After the EC9706 cells were exposed to 100 nmol/L bufalin,the number of cells gradually decreased in G(1) phase and increased in S and G(2)/M phases(p<0.05). The typical nucleus morphological changes of apoptosis were observed and the apoptotic index was increased(p<0.01). The expression of hTERT decreased in nucleus but increased in mitochondria(p<0.05).

CONCLUSIONSBufalin can inhibit the proliferation of human esophageal squamous carcinoma EC9706 cells in a time- and dose-dependent manner. It can arrest cell cycle in S and G(2)/M phases and induce the apoptosis of EC 9706 cells. hTERT is localized in both nucleus and mitochondria,and can be partially translocated from nucleus to mitochondria during the bufalin-induced apoptosis.

Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Telomerase ; metabolism

OBJECTIVETo investigate the clinicopathological value of the expression and amplification of P21-activated kinase 1 gene (PAK1) in colorectal carcinoma(CRC).

METHODSImmunohistochemistry (IHC), fluorescence in situ hybridization (FISH) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling(TUNEL) methods were used to examine the protein expression, amplification of PAK1 and cell apoptosis in 80 cases of CRC and 30 cases of colorectal adenoma by tissue microarray.

RESULTSIHC showed an overexpression of PAK1 protein in 26% of colorectal adenomas and 62% of CRCs. Significant association was found between expression of PAK1 and tumor histological grade as well as tumor clinical stage(P<0.05). In poor-differentiated(G(3)) CRCs, PAK1 expression in 90% carcinoma was up-regulated, which was significantly higher than that in tumors of G(1/2)(51%). Overexpression of PAK1 was detected in 78% of CRCs in later clinical stages (Dukes C, D), which was significantly higher than that in early clinical stages (Dukes A,B, 53%). In addition, negative correlation between PAK1 overexpression and cell apoptosis was observed in these CRC cohorts(P<0.05). FISH revealed that amplification of PAK1 gene was examined in only 3% CRCs.

CONCLUSIONSOverexpression of PAK1 protein may play an important role in development and progression of colorectal neoplasms and it is closely associated with the malignant histological and invasive phenotype of CRCs. The expression of PAK1 in CRC may be used as one of the new molecular markers in predicting tumors malignant potential and progression.

Apoptosis ; Colorectal Neoplasms ; genetics ; pathology ; Female ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Neoplasm Staging ; p21-Activated Kinases ; genetics

OBJECTIVECytomegalovirus (CMV) is the leading infectious cause of congenital anomalies of the central nervous system caused by intrauterine infection. However, the exact pathogenesis of these brain abnormalities has not been fully elucidated. It has been reported that periependymitis, periventricular necrosis and calcification are the most frequent findings in the brains of congenital CMV infection. Because a number of multipotential neural stem cells (NSCs) have been identified from ventricular zone, it is possible that NSCs in this area are primary targets for viral infection, which seems to be primarily responsible for the generation of the brain abnormalities. Therefore, the objective of the present study was to investigate the effect and mechanism of murine cytomegalovirus (MCMV) infection on neural stem cells' differentiation in vitro and its role in the mechanisms of brain abnormalities caused by congenital cytomegalovirus infection.

METHODSNSCs were prepared from fetal BALB/c mouse and were infected with recombinant MCMV RM461 inserted with a report gene LacZ at 1 multiplicity of infection (MOI = 1). The effect of MCMV infection on neural stem cells' differentiation was observed by detecting the ratio of nestin, GFAP and NSE positive cells with immunohistochemistry and flow cytometry on day 2 postinfection. The effects of MCMV infection on gene expression of Wnt-1 and neurogenin 1 (Ngn1) related to neural differentiation were detected by RT-PCR.

RESULTSNSCs isolated from embryonic mouse brains strongly expressed nestin, a specific marker of NSCs and had the capacity to differentiate into NF-200 and NSE positive neurons or GFAP positive astrocytes. At MOI = 1, the results of flow cytometry assay showed that nestin positive cells' proportion in the infection group [(62.2 +/- 1.8)%] was higher than that in the normal group [(37.2 +/- 2.4)%] (t = 4.62, P < 0.01). At the same time, the rates of GFAP and NSE positive cells' in the infection group were significantly lower than those in the normal group (P < 0.01). The scanning densities of Wnt-1 was 0.14 +/- 0.03 in the infection group while 0.32 +/- 0.04 in the control group (t = 7.21, P < 0.01). The scanning densities of Ngn1 were 0.09 +/- 0.01 and 0.21 +/- 0.02 in the two groups (t = 10.7, P < 0.01).

CONCLUSIONSThese results suggest that MCMV infection could inhibit neuronal differentiation, which may be primary causes of disorders of brain development in congenital CMV infection. The decreased expression of Wnt-1 and Ngn1 may be involved in the inhibitory effect of murine cytomegalovirus infection on neural stem cells' differentiation, which may lead to a new strategy for preventing and treating brain abnormalities caused by CMV infection through regulating these two signal pathways.

Animals ; Astrocytes ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Brain ; cytology ; Carrier Proteins ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Cytomegalovirus Infections ; congenital ; Embryo, Mammalian ; cytology ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Immunohistochemistry ; Intermediate Filament Proteins ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Multipotent Stem Cells ; metabolism ; virology ; Muromegalovirus ; Nerve Tissue Proteins ; genetics ; metabolism ; Nestin ; Neurons ; Pregnancy ; Reverse Transcriptase Polymerase Chain Reaction ; Wnt1 Protein ; genetics ; metabolism

Objective To investigate the pathogens of acute respiratory infection of children.Methods A total of 159 children with acute respiratory infection who were hospitalized in our department from August 2005 to January 2006 were involved in this study.The serum IgM antibody of 18 pathogens were detected by indirect immunofluorescence test.The 18 pathogens included respiratory syncytial virus(RSV),adenovirus(ADV),influenza A(H1N1,H3N2)and B viruses,parainfluenza viruses(PIV) type 1,2,3 and 4,coxsackie virus B1(CBV1),coxsackie virus A7(CAV7),echovirus(ECHO7),haemophilus influenzae(HI),klebsiella pneumoniae(KP),bordetella pertussis(BP),bordetella parapertussis(BPP) and legionella pneumophila serotype 1 and 12.Results The evidence of specific IgM was obtained in 103 of 159 patients(64.78%).Influenza A was found in 66 cases(64.08%),influenza B in 49 cases(47.57%),enterovirus in 26 cases(25.24%),RSV in 18 cases(17.48%),PIV in 11 cases(10.68%),and co-infection in 66 cases(64.08%),1/ 3 of them were co-infected with influenza A and B.Conclusions Viruses are the most common agents of acute respiratory infection.Influenza virus is predominant among them.

Pediatrics ; organization & administration ; Virology ; organization & administration

OBJECTIVEHuman rhinovirus (HRV) is the most common respiratory pathogen, which causes not only acute respiratory infection and community acquired pneumonitis in children, but also asthma episode and deterioration. However, the detection of respiratory pathogen, which mainly focuses on respiratory syncytial virus, influenzaviruses A and B, parainfluenza viruses 1-3 and adenoviruses, does not include HRV yet by now in China. The absence of detection method limits the clinical understanding of HRV pathogenicity, and causes unreasonable use of antibiotics. This study aimed to establish a one-step reverse transcription (RT) PCR system for detecting specific fragment of HRV RNA, and to analyze the sequences of amplicons.

METHODSA pair of degenerate primers based on the HRV highly conserved 5'' noncoding region (NCR) were used to develop a one-step RT-PCR system for detecting HRV RNA in nasopharyngeal aspirates from 78 children with acute respiratory tract infections in the spring of 2004. All the positive PCR products were sequenced, and the sequences of the nucleotides were analyzed by using biological software and compared with those in the GeneBank.

RESULTSEleven (14.1%) of 78 samples were positive on RT-PCR, these patients were clinically diagnozed as upper respiratory tract infection (n = 7), bronchitis (n = 3) and bronchopneumonia (n = 1), respectively. Compared with the sequences of clinical and standard HRV viruses in the GeneBank, the nucleotide sequences of these 11 amplicons shared high homology of 89%-95.5%. Within the 11 amplicons, nucleotide identity varied from 75.2% to 91.8%, and the ratio of genetic variation was from 8.8% to 31.0%, which occurred in highly conserved regions and usually showed single nucleotide mutation in some special locations. These 11 amplicons attribute to the two branches of HRV cladogram, respectively. Most of mutations in highly conservative domain occurred on single ribonucleotide, mainly as transversion (C/G, A/G) and transition (T/C, A/G), some were mutations among 3 bases (A/C/G, A/T/G, A/C/T). And a few mutations involved two nearby ribonucleotide which were also found in highly conservative domain. However, ribonucleotide deletion and insertion were usually found in highly variable domain.

CONCLUSIONThe findings showed that this one-step RT-PCR system was highly specific, rapid and convenient for the detection of HRV RNA in nasopharyngeal secretions of patients with acute respiratory tract infections and that the genome of HRV viruses was highly variable.

Base Sequence ; Child ; Child, Preschool ; Female ; Genes, Viral ; Humans ; Male ; Molecular Sequence Data ; Picornaviridae Infections ; diagnosis ; virology ; RNA, Viral ; analysis ; Respiratory Tract Infections ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Rhinovirus ; genetics ; isolation & purification ; Sensitivity and Specificity ; Sequence Analysis, DNA

BACKGROUNDThe production of neural stem cells (NSCs) derived from embryonic stem (ES) cells was usually very low according to previous studies, which was a major obstacle for meeting the needs of clinical application. This study aimed at investigating whether astrocytes could promote production of NSCs derived from ES cells in vitro.

METHODSMouse ES cells line-D3 was used to differentiate into NSCs with astrocytes as inducing stromal cells by means of three-stage differentiation procedure. Another group without astrocytes served as control. The totipotency of ES cells was identified by observation of cells' morphology and formation of teratoma in severe combined immunodeficiency disease (SCID) mice. The quantity and purity of NSCs derived from ES cells were analyzed using clonogenic assay, immunohistochemical staining and flow cytometry assay. The plasticity of NSCs was detected by differentiating test. Octamer-binding transcription factor 4 (Oct-4) and nestin, the specific marker genes of ES cells and NSCs respectively, were detected continuously using reverse transcription-polymerase chain reaction (RT-PCR) method to monitor the process of cell differentiation.

RESULTSThe ES cells of D3 line could maintain the ability of differentiating into cellular derivations of all three primary germ layers after continuous passage culture. At the end of two-stage of inducing process, 23.2 +/- 3.5 neurospheres per plate formed in astrocyte-induced group and only 0.8 +/- 0.3 per plate in the control group (clonogenic assay, P < 0.01), and the ratio of nestin positive cells was (50.2 +/- 2.8)% in astrocyte-induced group and only (1.4 +/- 0.5)% in the control group (flow cytometry, P < 0.01). With the induction undergoing, the expression of Oct-4 gradually decreased and then disappeared, while the expression of nestin was increased step by step, and the ratio of nestin positive cells was up to 91.4% by the three-stage differentiation. The nestin positive cells could be further induced into neurons, astrocytes, and oligodendrocytes in differentiating medium supplemented with fetal calf serum. The results of differentiating test showed that the ratio of NF-200 and NSE positive cells was (42.7 +/- 2.6)% in astrocyte-induced group and only (11.2 +/- 1.8)% in the control group (P < 0.01).

CONCLUSIONSAstrocytes can not only increase the production of NSCs derived from ES cells but also promote the differentiation of NSCs toward neuronal lineage.

Animals ; Astrocytes ; physiology ; Cell Differentiation ; Cell Lineage ; Cells, Cultured ; Embryo, Mammalian ; cytology ; Mice ; Neurons ; cytology ; Stem Cells ; cytology

Background The production of neural stem cells (NSCs) derived from embryonic stem (ES) cells was usually very low according to previous studies, which was a major obstacle for meeting the needs of clinical application. This study aimed at investigating whether astrocytes could promote production of NSCs derived from ES cells in vitro.Methods Mouse ES cells line-D3 was used to differentiate into NSCs with astrocytes as inducing stromal cells by means of three-stage differentiation procedure. Another group without astrocytes served as control. The totipotency of ES cells was identified by observation of cells' morphology and formation of teratoma in severe combined immunodeficiency disease (SCID) mice. The quantity and purity of NSCs derived from ES cells were analyzed using clonogenic assay, immunohistochemical staining and flow cytometry assay. The plasticity of NSCs was detected by differentiating test. Octamer-binding transcription factor 4 (Oct-4) and nestin, the specific marker genes of ES cells and NSCs respectively, were detected continuously using reverse transcription-polymerase chain reaction (RT-PCR) method to monitor the process of cell differentiation. Results The ES cells of D3 line could maintain the ability of differentiating into cellular derivations of all three primary germ layers after continuous passage culture. At the end of two-stage of inducing process, 23.2±3.5 neurospheres per plate formed in astrocyte-induced group and only 0.8±0.3 per plate in the control group (clonogenic assay, P<0.01), and the ratio of nestin positive cells was (50.2±2.8)% in astrocyte-induced group and only (1.4±0.5)% in the control group (flow cytometry, P<0.01). With the induction undergoing, the expression of Oct-4 gradually decreased and then disappeared, while the expression of nestin was increased step by step, and the ratio of nestin positive cells was up to 91.4% by the three-stage differentiation. The nestin positive cells could be further induced into neurons, astrocytes, and oligodendrocytes in differentiating medium supplemented with fetal calf serum. The results of differentiating test showed that the ratio of NF-200 and NSE positive cells was (42.7±2.6)% in astrocyte-induced group and only (11.2±1.8)% in the control group (P<0.01).Conclusions Astrocytes can not only increase the production of NSCs derived from ES cells but also promote the differentiation of NSCs toward neuronal lineage.

Adrenal Cortex Hormones ; therapeutic use ; Autoantigens ; immunology ; Child ; Cytokines ; metabolism ; HLA Antigens ; genetics ; Hepatitis, Autoimmune ; diagnosis ; immunology ; therapy ; Humans ; Immunosuppressive Agents ; therapeutic use ; Liver ; drug effects ; metabolism ; pathology ; Liver Transplantation