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

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

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

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 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 study the effect of transfecting survivin antisense mRNA on growth and chemotherapy sensitivity of lymphoma cells.

METHODSEukaryotic expression plasmid pcDNA3. 1-antisense (As) survivin was constructed and transfected into Jurkat T lymphoblastic lymphoma cell lines with high expression survivin mRNA by use of lipofectmine gene transfer technique. Expression of survivin mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemical and Western blot. The effect of transfecting survivin antisense mRNA on the growth of Jurkat cell lines was monitored by population doubling time (PDT) and Apoptotic indexes (AI). The morphologic features were observed in transfected cells by light and electric microscopes. MTT assay was used to analyze the response of transfected cells to CTX and MTX.

RESULTSCompared with the control cells, the expression of survivin mRNA and protein were reduced after transfected pcDNA3. 1-Assurvivin 48 h, 5 w and 6 w, PDT (52 h) was prolonged. Apoptotic indexes were higher in transfected antisense survivin mRNA cells [20.2% (48 h)], 6.2% (5 w) and 6.8% (6 w) than control ones [2.1%, 1.3% (48 h)] and [1.3% (5 w) and 1.0% (6 w)]. The cells grow slowly and the dead cells increase and some swelling and apoptotic cells were observed in transfected pcDNA3. 1-Assurvivin groups by invert, light and electric microscopes. The Jurkat cell line of transfected pcDNA3. 1-Assurvivin had higher sensitivity to CTX and MTX. The rate of inhibition was higher in transfected group. There is a significant difference between the transfected group and untransfected one, P < 0.05.

CONCLUSIONSThe result indicated that survivin gene was very important for growth of Jurkat cells. To inhibit the expression of survivin will be significant in therapy of T lymphoblastic lymphoma. Survivin gene might be a target of therapy.

Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents, Alkylating ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cyclophosphamide ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; Jurkat Cells ; cytology ; metabolism ; K562 Cells ; cytology ; metabolism ; Lymphoma ; pathology ; Methotrexate ; pharmacology ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Neoplasm Proteins ; biosynthesis ; genetics ; Plasmids ; RNA, Antisense ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

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 analyze the possible causes of BPH surgery-related urethral stricture and summarize the experience in its clinical management.

METHODSThe clinical data of 37 cases of BPH surgery-related urethral stricture were analyzed retrospectively. The patients averaged 68.5 years in age, of whom 12 had the history of open surgery, and 25 transurethral surgery. Anterior urethral stricture was found in 6 cases, and posterior urethra in stricture 31. Thirty-five cases were treated by holmium laser urethrotomy (HLU) or a combination of HLU with transurethral resection of the scar in the stenotic segment, and the results were evaluated based on the maximum flow rate (Qmax).

RESULTSTwo cases of full penile urethral stricture were treated by urinary diversion, and all the rest by urethral stricture surgery. Catheters were indwelt in 4 cases of urethral stenosis for 5-7 days post-operatively, and smooth urination was achieved after their removal, with a Qmax of > 15 ml/s. Another 31 cases of membranous urethral stricture received catheter indwelling of 2 -4 weeks and were followed up for 1 - 21 (mean 11. 5) months, of whom 23 experienced satisfactory voiding, with an average Qmax of 14.3 ml/s, and the other 8 poor voiding, with a Qmax of < 8 ml/s.

CONCLUSIONHLU or a combination of HLU with transurethral resection of the scar in the stenotic segment is an effective method for the treatment of BPH surgery-related urethral stricture. And conscientiously performed BPH surgery can reduce the incidence of urethral stricture.

Aged ; Humans ; Laser Therapy ; Lasers, Solid-State ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Prostatic Hyperplasia ; surgery ; Retrospective Studies ; Urethra ; Urethral Stricture ; etiology ; prevention & control