Acrosin ; immunology ; isolation & purification ; Humans ; Male ; Membrane Proteins ; immunology ; isolation & purification ; Protein Binding ; Spermatozoa ; metabolism ; Zona Pellucida ; metabolism

BACKGROUND:At present, there are many researches about repairing articular cartilage defects. In particular, the microfracture technique has been widely used. OBJECTIVE:To observe recovery of knee joint motor function and morphological changes in tissue repair during articular cartilage defects with different directions (coronal position and sagittal position). METHODS:Articular cartilage fracture models with 2 mm-thick medial femoral condyles of rabbit knee joint were established. According to incision directions, models were assigned to coronal and sagittal groups. At 5, 10 and 20 weeks after model induction, general observation was performed. Specimens were sliced into paraffin sections, and subjected to hematoxylin-eosin staining and col agen staining. Tissue repair at the articular cartilage defects was observed using optical microscope and immunohistochemical method. After model induction, range of motion of rabbit joints was regularly examined in the two groups.RESULTS AND CONCLUSION:A white line was seen across the femoral condyles at defects in the two groups. Articular surface at defect repair was at the level of in situ cartilage, and reached a bone union. Knee joint treated by operation did not affect function. Under light microscope, partial reconstruction of subchondral bone was seen in the two groups, mainly fibrocartilage repair. The level of bony remodeling was lower than tidal line of adjacent in situ cartilage. Immunohistochemical method exhibited that type I col agen staining gradual y reduced at defects of specimens, but type II col agen staining gradual y increased. These results suggested that there was no significant difference in the recovery of motor function of knee joint and the repair of articular cartilage with different directions (coronal and sagittal position).

Objective: To explore the levels of IL-22 in thymus damaged by γ-ray total body irradiation (TBI), and to study the role of IL-22 in T cell reconstitution after thymic injury induced by TBI. Methods: To induce thymic injury, mice were treated by sub-lethal TBI. Levels of intra-thymic and circulatory IL-22 were detected by using ELISA assay. Untreated mice were used as control. After receiving sub-lethal TBI, mice were intraperitoneally injected with PBS or recombinant mouse IL-22, which were marked as TBI+PBS or TBI+IL-22, respectively. Mice were monitored for counts of total thymic cells and circulatory white blood cells. Flow cytometry was applied to analyze percentages of thymic epithelial cells (TEC), thymocyte subsets and circulatory T cells. Real-time PCR assay was applied to analyze the mRNA expression levels of Foxn1, Ccl25, Aire and Dll4 in thymus. Results: ①Sub-lethal TBI treated mice expressed higher levels of intra-thymic and circulatory IL-22, compared with untreated ones (all P<0.05). ②After injection of recombinant IL-22, TBI+IL-22 mice had higher levels of intra-thymic IL-22 than TBI+PBS mice (all P<0.05). ③On day 14 after irradiation, real-time PCR assay showed that TBI+IL-22 mice had higher mRNA levels of Foxn1, Ccl25, Aire and Dll4 in thymus compared with TBI+PBS ones. Meanwhile, the TBI+IL-22 mice had higher counts of total thymic cells[(5.93±3.19)×10(6)/ml vs (1.42±0.46)×10(6)/ml, t=3.128, P=0.033] and circulatory white blood cells[(3.08±0.94)×10(6)/ml vs (1.43±0.30)×10(6)/ml, t=3.730, P=0.015] than those of TBI+PBS mice. Flow cytometry analysis indicated that TBI+IL-22 mice had higher counts of TEC and thymocytes than TBI+PBS mice on day 14 after irradiation (all P<0.05). On days 7 and 14 after irradiation, TBI+IL-22 mice had higher counts of circulatory white blood cells and T cells than TBI+PBS mice (all P<0.05). Conclusion: Sub-lethal TBI induces upregulation of intra-thymic IL-22, and injecting of recombinant IL-22 increases level of IL-22 in thymus. Injecting of recombinant IL-22 improves recovery of TEC and increases numbers of thymocyte subsets and circulatory T cell after thymic injury.
Animals ; Interleukins ; Mice ; Radiation, Ionizing ; T-Lymphocytes ; Thymus Gland ; Whole-Body Irradiation ; Interleukin-22

To generate transgenic mice in which both hygromycin (hyg) and neomycin (neo) resistance genes are expressed in murine fibroblast cells (MEFs), which are required for conditional gene knock-out and screening of drug resistant ES cell clones. To construct HygR-neoR expression vector, pTK-hygR-pA and PGK-neoR-pA were cloned into pBluescript vector. DNA fragments of tandem genes ( 4245bp ) were prepared by Kpn I and Xba I digestion and transgene was microinjected into pronucleus of zygotes to generate transgenic mice. Transgenic mice were identified by PCR and Southern blot; expression of hygR and neoR gene transcripts were detected by RT-PCR. 7 founder mice carrying hyg-neo resistant genes were obtained and 6 transgenic mouse lines were successfully established. The hygR and neoR gene transcripts were detected in the liver and/or ovary of transgenic mice from hn30, hn33, hn66 and hn67 mouse lines. In MEFs isolated from the mice of line hn66 and hn30, expression of hyg and neo resistant genes was also detectable. Transgenic mouse lines expressing two anti-drug genes have been established. The hyg and neo resistant gene transcripts were detected in the MEFs of two transgenic mouse lines.
Animals ; Cinnamates ; pharmacology ; Drug Resistance, Multiple ; genetics ; Fibroblasts ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mice ; Mice, Transgenic ; Neomycin ; pharmacology ; Transgenes ; genetics

OBJECTIVETo investigate the morphology of the soft palate in normal individuals with digital radiography, when they pronounced the high vowel of "i", and to provide the references for therapy of the cleft palate.

METHODSIn this study, the sample comprised 27 normal subjects. With the digital cephalometry, the morphology of the soft palate when pronouncing the high vowel of "i" was observed. And the dimensional difference of the soft palate when pronouncing between different gender was studied.

RESULTSWhen pronouncing the high vowel of "i", the morphology of the soft palate was like the shape of the knee. And it could be divided into two parts: horizontal and vertical. The length of the vertical part in male group was (24.92 +/- 2.03) mm, the length of the vertical part in the female group was (20.66 +/- 2.77) mm. The length of the vertical part was different between male and female group (P < 0.001).

CONCLUSIONThe morphology of the palate when pronouncing the high vowel of "i" is similar. And the velar length of the vertical part of the male is longer than the female.

Cephalometry ; Cleft Palate ; Female ; Humans ; Male ; Palate, Soft

BACKGROUNDInfusion phlebitis is the most common side effect of clinical intravenous drug therapy and several clinical studies have demonstrated that anisodamine can effectively prevent the occurrence of infusion phlebitis. This study was designed to investigate effects of anisodamine on the expressions of vascular endothelial growth factor (VEGF) and intercellular adhesion molecule 1 (ICAM-1) in a rabbit model of infusion phlebitis and to analyze the mechanisms of anisodamine effect on the prevention and treatment of experimental infusion phlebitis.

METHODSTwenty-four specific pathogen-free male Japanese white rabbits were randomly assigned to the control group, the model group, the magnesium sulfate group and the anisodamine group. The rabbit model of infusion phlebitis, induced by intravenous administration, was established and expressions of VEGF and ICAM-1 were determined and contrasted with the control group treated with normal saline. We evaluated expression by histopathology, immunohistochemistry, reverse transcription-polymerase chain reaction, and Western blotting assay.

RESULTSPathohistological changes of the model group were observed, such as loss of venous endothelial cells, inflammatory cell infiltration, edema and thrombus. The magnesium sulfate group and the anisodamine group showed significant protective effects on vascular congestion, inflammatory cell infiltration, proliferation, swelling of endothelium and perivascular hemorrhage. The model group showed the highest expressions of VEGF and ICAM-1 of the four groups (P < 0.01). On the contrary, anisodamine alleviated the inflammatory damage by significantly reducing the expressions of VEGF and ICAM-1 compared with the model group (P < 0.01). There was no significant difference in the expressions of VEGF and ICAM-1 between the magnesium sulfate group and the anisodamine group (P > 0.05).

CONCLUSIONAnisodamine alleviates inflammatory damage by significantly reducing the expressions of VEGF and ICAM-1, and shows significant protective effects in an animal model of infusion phlebitis.

Animals ; Blotting, Western ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Phlebitis ; drug therapy ; Rabbits ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Solanaceous Alkaloids ; therapeutic use ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo establish a nasopharyngeal carcinoma (NPC) cell line with stable EIF4G1 gene silencing induced by small interfering RNA (siRNA).

METHODSThe EIF4G1 mRNA levels in 8 NPC cell lines including 5-8F, 6-10B, C666-1, CNE1, CNE2, HNE1, HONE1, and SUNE1 were detected by fluorescence quantitative RT-PCR (QRT-PCR). The recombinant lentivirus shRNA expression plasmid targeting EIF4G1 gene was packaged into mature lentivirus by 293FT cells and used to infect 5-8F cells. After blasticidin selection of NPC cells with constant expression of the EIF4G1-siRNA, the efficiency of EIF4G1 mRNA expression interference was determined using QRT-PCR.

RESULTSThe 8 NPC cell lines showed differential expression of EIF4G1 mRNA, among which 5-8F cells had the highest EIF4G1 expression. The recombinant lentivirus plasmid pLenti6/BLOCK-iT-DEST/EIF4G1-shRNA was successfully constructed and verified by PCR and sequencing. The EIF4G1 mRNA level of 5-8F cells infected with shRNA-EIF4G1 lentivirus was significantly reduced as compared with the negative control and the blank control cells.

CONCLUSIONThe recombinant lentivirus vector pLenti6/BLOCK- iT-DEST/EIF4G1-shRNA we constructed results in marked downregulation of EIF4G1 mRNA expression and constant expression of EIF4G1-siRNA after infection of 5-8F cells.

Cell Line, Tumor ; Eukaryotic Initiation Factor-4G ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo investigate the usefulness of electromyogram (EMG) in the diagnosis of the patients with hemimasticatory spasm (HMS).

METHODSFour cases with HMS were reported. All the 4 patients were undertaken needle and surface electrode EMG examination.

RESULTSNeedle electrode EMG of the 4 patients with HMS showed grouped potentials synchronously with the onset of the spasm, which indicated abnormal excitatory electrical activities of the trigeminal nerve resulting in involuntary masticatory muscle movements.

CONCLUSIONIt is very important to use EMG for the diagnosis of HMS.

Adult ; Diagnosis, Differential ; Electromyography ; Female ; Humans ; Male ; Masticatory Muscles ; physiopathology ; Middle Aged ; Spasm ; diagnosis ; etiology ; physiopathology

OBJECTIVETo compare the synthetic ability of osteoblasts on the surface of different nano-granule titanium films and investigate the correlation between nanophase titanium films and cellular biocompatibility.

METHODSFour different nano-granule titanium films were produced by direct current magnetron sputtering, at ambient, 100 degrees C, 250 degrees C, 380 degrees C substrate temperature, respectively. Rat osteoblasts were seeded on the surface of four treated groups of titanium film samples and non-treated Ti sample(control group). The production of osteocalcin (OC) in all five groups were detected by using double antibody sandwich enzyme-linked immunosorbent assay.

RESULTSThe production of OC increased gradually from day 7 to day 14 in all groups. In the control group, it showed significant differences with other five groups on day 7. On day 14, the production of OC in 100 degrees C group was the highest, and it showed significant differences with 380 degrees C, control group and blank group. In 250 degrees C group, the production of OC also showed significant differences with 380 degrees C, control group and blank group (P < 0.05).

CONCLUSIONTitanium with nano-modified surface had good biocompatibility and different nano-granule titanium films could affect the synthesis of osteoblasts.

Animals ; Osteoblasts ; Osteocalcin ; Rats ; Surface Properties ; Titanium

OBJECTIVETo examine the distribution of fluorouracil in gastric cancer (CA), lymph node (LN), normal gastric mucosa (NG), peritoneum (PE), greater omentum (GO) and lesser omentum (LO) by preoperative intraperitoneal chemotherapy with Co-fluorouracil liposome (Co 5-Fu), and offer an experimental basis for clinic practice.

METHODSNinety-six gastric cancer patients were divided into four groups: Co 5-Fu i.v. injection group (Co 5-Fu i.v.), Co 5-Fu intraperitoneal perfusion group (Co 5-Fu i.p.), 5-Fu i.v. injection group (5-Fu i.v.) and intraperitoneal perfusion group (5-Fu i.p.) given on day-2, day-1 and 60 minutes before operation. Fluorouracil concentration in all tissues collected during operation were examined by high performance liquid chromatography (HPLC).

RESULTSThe fluorouracil concentration in the tissues in Co 5-Fu i.p. group was significantly higher than that in Co 5-Fu i.v. or 5-Fu i.p. group (P < 0.05 or P < 0.01), and that in 5-Fu i.p. group was greatly higher than that at 5-Fu i.v. group (P < 0.01). In Co 5-Fu i.p. group, the concentration of drug in LN, CA, PE, NG, GO and LO decreased gradually with the former 3 tissues significantly higher than the latter 3 tissues (P < 0.01), and adjacent lymph node was the highest. In Co 5-Fu i.v. group, the ranking was LN, CA, NG, PE, GO and LO with the former 3 tissues significantly higher than the latter 3 tissues (P < 0.01) and showing tumor tissues higher than the other tissues (P < 0.01). In 5-Fu i.p. group, the ranking was PE, LN, CA, NG, GO and LO with the former 2 tissues significantly higher than the latter tissues (P < 0.01).

CONCLUSIONCo 5-Fu possesses drug targeting, slow release and long effect in gastric cancer tissues and adjacent lymph nodes. Preoperative chemotherapy with Co 5-Fu i.p. is more advantageous than 5-Fu given i.v. or 5-Fu i.p.

Aged ; Antimetabolites, Antineoplastic ; administration & dosage ; pharmacokinetics ; Female ; Fluorouracil ; administration & dosage ; pharmacokinetics ; Gastric Mucosa ; metabolism ; Humans ; Infusions, Parenteral ; Injections, Intravenous ; Liposomes ; Lymph Nodes ; metabolism ; Male ; Middle Aged ; Omentum ; metabolism ; Panax ; chemistry ; Peritoneum ; metabolism ; Polysaccharides ; administration & dosage ; isolation & purification ; pharmacokinetics ; Preoperative Care ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology