Actins ; metabolism ; Carcinoembryonic Antigen ; metabolism ; Desmin ; metabolism ; Diagnosis, Differential ; Epithelial Cells ; metabolism ; pathology ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neoplasms, Complex and Mixed ; metabolism ; pathology ; surgery ; Neoplasms, Glandular and Epithelial ; metabolism ; pathology ; surgery ; Stromal Cells ; metabolism ; pathology ; Vimentin ; metabolism

Diagnosis, Differential ; Endothelium, Vascular ; metabolism ; pathology ; Female ; Humans ; Hyperplasia ; Jejunal Diseases ; metabolism ; pathology ; surgery ; Jejunum ; blood supply ; pathology ; surgery ; Middle Aged ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism

Humans ; Liver Neoplasms ; pathology ; Perivascular Epithelioid Cell Neoplasms ; pathology

Actins ; metabolism ; Adult ; Desmin ; metabolism ; Electromyography ; Humans ; Male ; Microscopy, Electron, Transmission ; Muscle, Skeletal ; pathology ; ultrastructure ; Myopathies, Nemaline ; metabolism ; pathology

To investigate the antitumor immune responses induced by MAGE-3 DNA vaccine, the recombinant mammalian expression plasmid pcDNA3.1/MAGE-3 was constructed by ligating MAGE-3 gene, which was amplified by RT-PCR, and the pcD-NA3.1 + vector. The recombinant plasmids were transfected into B16 cells by liposome, the expression of MAGE-3 was checked by RT-PCR, immunocytochemistry and Western blot. Then, 100 ug recombinant plasmids were injected intramuscularly per C57BL/6 mouse on 0, 10 and 20 days, with pcDNA3.1 + plasmid and PBS as controls. Splenocytes CTLs, the level of antibodies against MAGE-3 the changes of the T lymphocyte subsets and the levels of cytokines were checked after 3 times immunization. As a result, the mice immunized with pcDNA3.1/MAGE-3 plasmid can produce MAGE-3 specific immune response. The CTLs kill activities against B16/MAGE-3 cells was 51.08 +/- 7.41%, and had significant difference (P < 0.01) compared with that of pcDNA3.1 + group (8.44 +/- 1.89%) and PBS group (5.76 +/- 1.75%). The titre of antibody against MAGE-3 was 1:15, while controls were negtive. The number of CD4 + CD8 + and the levels of IFN-gamma IL-2 increased significantly after immunization with pcDNA3.1/MAGE-3 plasmid as compared with those of control groups (P < 0.01). It is concluded that the pcDNA3.1-MAGE-3 DNA vaccine are able to induce both cellular and humoral immune responses in vivo.
Animals ; Antibodies, Neoplasm ; blood ; Antigens, Neoplasm ; biosynthesis ; genetics ; immunology ; Cancer Vaccines ; biosynthesis ; immunology ; Female ; Melanoma, Experimental ; prevention & control ; Mice ; Mice, Inbred C57BL ; Neoplasm Proteins ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; T-Lymphocyte Subsets ; immunology ; Vaccination ; Vaccines, DNA ; biosynthesis ; genetics ; immunology

To express the GST-MAGE-3 protein in E. coli, and investigate the antitumor immune responses induced by Dendritic cells (DCs) pulsed with GST-MAGE-3 protein, the recombinant expression plasmid pGEX-MAGE-3 was constructed by ligating MAGE-3 gene, which was amplified by RT-PCR and confirmed by sequencing, and the pGEX-4T-2 vector. The recombinant plasmid was transformed into BL-21 E. coli. The expression of GST-MAGE-3 was induced with IPTG. The GST-MAGE-3 protein expressed as high as 32% of the total cellular protein. After purification with Glutathione Sepharose 4B, the purity of the protein was more than 90%, and 3mg GST-MAGE-3 was obtained from 100 mL BL-21 lysate. Dendritic cells from gastric carcinoma patients were pulsed with GST-MAGE-3 protein, and these DCs were used to stimulate the autologous T. lymphocytes. After 7 days, the T. lymphocytes cocultured with DCs pulsed with GST-MAGE-3 antigen exhibited specific cytotoxicity against MAGE-3 positive SGC-7901 cells. It is concluded that the GST-MAGE-3 protein are able to present antigen to T. lymphocytes, activate antigen-specific CTLs and induce special antitumor immune responses in vitro. Our results lay the groundwork for further research of the MAGE-3 vaccine.
Antigens, Neoplasm ; genetics ; isolation & purification ; metabolism ; pharmacology ; Blotting, Western ; Cell Line, Tumor ; Cells, Cultured ; Dendritic Cells ; drug effects ; immunology ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Neoplasm Proteins ; genetics ; isolation & purification ; metabolism ; pharmacology ; Plasmids ; genetics ; Stomach Neoplasms ; immunology ; T-Lymphocytes ; immunology

OBJECTIVETo detect the self-assembly morphology of sodium hyaluronate injection on mica using atomic force microscopy(AFM).

METHODSAtomic force microscopy with nanometer resolution was used to observe the self-assembly morphology of different concentrations of sodium hyaluronate injection on mica at room temperature.

RESULTSThe self-assembly morphology of 0.001, 0.01, and 0.1 mg/ml sodium hyaluronate injection on mica featured piebald, reticular and dendritic structures, respectively. At 1 and 5 mg/ml, sodium hyaluronate injection displayed bacilliform and spherical structures on mica, respectively; the diameter and height of the particles of 5 mg/ml sodium hyaluronate was 197.97±78.48 nm and 30.79±18.67 nm, significantly greater than those of 0.1 mg/ml sodium hyaluronate injection (49.52±11.93 nm and 5.37±1.59 nm, respectively, P<0.05).

CONCLUSIONThe self-assembly morphology of sodium hyaluronate injection on mica varies with its concentration. The piebald and reticular structure may facilitate the function of sodium hyaluronate, and the dendritic feature resembles the representative model of diffusion-limited aggregation (DLA).

OBJECTIVETo investigate the injury in the corneal nerve and cornea of rats exposed to n-hexane.

METHODSThirty-two SD male rats were randomly divided into one control group and four n-hexane groups. The four n-hexane groups inhaled 35.2 g/m(3) n-hexane statically for 1, 3, 7 and 14 d respectively, while the rats in the control group inhaled air. The corneal nerve damage was investigated with golden staining and transmission electron microscope. Histopathological and ultrastructure changes of cornea were analyzed also.

RESULTSThe concentration of n-hexane in blood of rats in different experimental groups was (242.91 +/- 59.68), (668.77 +/- 221.74), (1021.21 +/- 545.71) and (1140.42 +/- 468.44) microg/L, increased gradiently with time exposed to n-hexane. In the rats exposed to n-hexane for 7 and 14 d, there appeared fewer corneal nerve bundles and lower density of nerve fiber at the center of cornea, under electron microscope, the lamellar sheath of nerve fiber in the corneal epitheliums appeared intermittent, the neuroplasm of endings was partly lysed and became vacuolar, the microfilament and racuole of neuraxon decreased. In the group exposed to n-hexane for 14 d, the microvillus of cornea epithelium were decreased. In some basal cells there appeared pyknotic nucleus and vacuole, mitochondria were swollen or disappeared.

CONCLUSIONThe structure of corneal nerve and cornea is damaged in the rats exposed to n-hexane, thus leading to dysfunction of cornea.

Objective To explore the biological functions of retinoic acid-inducible gene-I(RIG-I) in vivo through phenotype analysis of RIG-I knockout mice. Methods The gene expression of RIG-Ⅰ in various tissues of mice was examined with Northern blotting and semi-quantitative RT-PCR.The phenotypes observed included body weight measurement,differential count of peripheral blood cells,metabolic parameters measurement and histopathologic examination. ResultsRIG-Ⅰ expressed in various tissues of mice with different levels.No gross developmental abnormalities and expected maturation arrest in granulocytic differentiation were observed in RIG-Ⅰ knockout mice.However,RIG-Ⅰ knockout mice exhibited an unexpected increase in the ratios of neutrophiles to lymphocytes in peripheral blood and increased susceptibility to bacteria infection. Conclusion RIG-Ⅰ may play an important role in immune regulation in mice.

Objective:To investigate the predictive value of cerebroplacental ratio (CPR) for adverse perinatal outcomes of induction of labor in prolonged pregnancy.Methods:This retrospective study recruited 315 singleton pregnant women who had induced labor due to prolonged pregnancy (≥41 gestational weeks) in the First Affiliated Hospital of Chongqing Medical University from January 1, 2019 to April 30, 2020. Based on the occurrence of adverse perinatal outcomes (emergency delivery due to persistent abnormal fetal heart rate monitoring, umbilical artery blood pH at birth <7.2, 5 min Apgar scores<7, transferring to neonatal intensive care unit after birth, chorioamnionitis and vaginal delivery converted to cesarean section), they were divided into two groups: case group ( n=76) and normal group ( n=239). Clinical features and umbilical artery blood flow, middle cerebral artery (MCA) flow and CPR measured in the last ultrasound scan before induction were compared between the two groups using student's t-test, Mann-Whitney U test and Chi-square test. Receiver operating characteristic (ROC) curve was used to analyze the predictive values of umbilical artery blood flow, MCA flow and CPR for the adverse perinatal outcomes. Multivariate logistic regression analysis was used to screen the meaningful predictors. Results:Compared with the normal group, the umbilical artery pulsatility index (PI) (0.9±0.1 vs 0.8±0.1, t=-5.458, P<0.001) and the percentage of abnormal CPR (<1.0) increased significantly [21.1%(16/76) vs 6.3%(15/239), χ2=14.190, P<0.001] in the case group, while the MCA-PI and CPR decreased significantly (1.1±0.2 vs 1.3±0.3, t=5.658, P<0.001; 1.2±0.3 vs 1.6±0.5, t=8.940, P<0.001). The areas under the ROC curves of umbilical artery PI, MCA-PI and CPR for predicting adverse perinatal outcomes were 0.71, 0.71 and 0.77, respectively. CPR had the highest sensitivity (0.74) compared with umbilical artery PI (0.68) and MCA-PI (0.71), but the specificity of them were similar (0.67, 0.66 and 0.66). Multivariate logistic regression analysis showed that only CPR was the independent risk factor for adverse perinatal outcomes ( OR=0.028, 95% CI: 0.010-0.080, P<0.001). Conclusions:As an indicator for early prediction of adverse perinatal outcomes of induction of labor in prolonged pregnancy, CPR was more sensitive but less specific.