Animals ; Cells, Cultured ; Chondrocytes ; Interleukin-1beta ; NF-kappa B ; Osteoarthritis/drug therapy* ; Rats ; Tumor Necrosis Factor-alpha

Objective To investigate the relationship of impact stress distribution and stress wave dissemination with temporal bone fracture. Methods Twelve donated fresh corpse were selected and impacted with driving force of 800 kPa, 1 200 kPa and 1 400 kPa to establish the model of impact temporal bone fractures. With aid of three-dimensional stereochemical structure image of 64-slice CT, the vitodynamic distribution and dissemination of impacted temporal bone was calculated by using computer simulation. Results (1) The instant velocity, acceleration and energy of impact as well as displace-ment scope of the hammer increased with higher driving force in the tempus. (2) Instantaneous impact of the temporal bone resulted in Von Mises stress concentration area around the impact point. The stress was weakened suddenly in the skull-brain tissue coupling place but later accumulated in the skull base. (3) Only ipsilateral linear fracture occurred when the driving force of impact was 800 kPa. While impact dif-ferent degrees of depressed fractures could be seen and one case showed secondary fracture of the opposite side when the driving force of impact was 1 200 kPa. The impact with driving force for 1 400 kPa begot complex ipsilateral fractures, secondary fracture of the opposite side and skull basal fracture. (4) Frac-ture line of the temporal bone accorded with impact stress concentration area. Conclusions All pa-rameters, stress distribution and dissemination of impact are in accordance with temporal bone fracture during impaet process of the temporal bone, which is of vital significance for diagnosis and protection of impact temporal bone fractures.

ObjectiveTo observe the injured changes of brain myelin sheath structure and myelin basic protein (MBP) content induced by amyloid β peptide (Aβ) and effect of GETO on these changes.MethodsThe experimental rat model of Alzheimer's disease was established with Aβ1-42 injection into hippocampus. 4 weeks later, the myelin sheath structure of the CA1 area of the rat hippocampus was taken and observed by electromicroscope, and distribution and content of MBP were examined with immunohistochemical method.ResultsThe electromicroscope showed that the structure of myelin sheath became relaxing, disorder, homogenization and default of hippocampus CA1 in the model rats. In GETO treated group, the structure of myelin sheath was integrity and continuum. Immunohistochemical test showed that the staining and numbers of myelin sheath of model rats was thinner than that of normal rats and GETO treated rats. The numbers, mean area and mean density of positive staining axon in hippocampus CA1 of MBP in the model rats were significantly different from those in the normal group and GETO group (P<0.01).ConclusionAβ1-42 injection into hippocampus in rats can impair myelin sheath to make MBP release and GETO can ameliorate these changes.

Objective To clone the novel activation-related gene of B lymphocyte. Methods The differential display reversal transcription PCR (DDRT-PCR) technique was applied to analyse the expression difference of mRNA between resting and activated B lymphocyte from human tonsil. The positive differential display cDNA fragment identified by Northern-blotting was chosen as probe to filtrate human activated B lymphocyte cDNA library. Results Sixty two differential display cDNA fragments(expressed sequence tag, EST)were obtained. Thirty-two of them were mainly expressed in resting B lymphocyts and thirty were expressed in activated cells. Twenty-five were positive ones after identification by Northern blot analysis. A novel cDNA clone was obtained after using EST30 as a probe to filtrate the human activated B cell cDNA library.The whole cDNA clone was 2 048 bp in length and contains a 630 bp open reading frame. The N end of the deduced amino acid sequence was homologous with KAR3 protein which is a member of kinesins superfamily in yeast. Conclusions A novel possible activation-related gene in human B lymphocyte was obtained.

OBJECTIVETo observe microvessel density(MVD), epithelial stromal vascular cuffing(VC) and expression of vascular endothelial growth factor(VEGF) in human cervical carcinomas and to clarify their significance in the invasion and metastasis of cervical carcinoma.

METHODSVEGF and CD34 were stained immunohistochemically (SP) in 57 cases of cervical carcinoma (30 cases of squamous cell carcinoma, 20 of adenocarcinoma 7 of glandular and squamous cell carcinoma), 29 cases of cervical intraepithelial neoplasia (CIN) and 16 cases of normal cervices, meanwhile, MVD and VC were also assayed.

RESULTSThere were significant differences among the above 5 groups for MVD P<0.01 . The VC pattern showed a significant difference between cervical carcinoma and CIN or control group P<0.01). The positive rates of VEGF in normal cervical epithelium, CIN, squamous cell carcinoma, adenocarcinoma, glandular and squamous cell carcinoma were 18.8% 3/16, 82.8% 24/29), 93.3% 28/30), 100% 20/20 and 7/7(100%), respectively. There were significant differences between these cervical lesion groups and the control group(P<0.001). The MVD showed significant differences between the positive pelvic node metastasis and negative pelvic node metastasis P<0.05). There was no significant correlation between the expression of VEGF and the tumor diameter, clinical stage, pathologic grade and pelvic node metastasis.

CONCLUSIONThe expression of VEGF may play an important role in the angiogenesis of cervical carcinoma. Degree of malignancy of cervical carcinoma has a close association with microvessel density.

Adult ; Aged ; Aged, 80 and over ; Endothelial Growth Factors ; analysis ; Female ; Humans ; Intercellular Signaling Peptides and Proteins ; analysis ; Lymphatic Metastasis ; Lymphokines ; analysis ; Microcirculation ; Middle Aged ; Uterine Cervical Neoplasms ; blood supply ; chemistry ; pathology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Objective To investigate the polymorphism of human cytomegalovirus（HCMV）UL146 gene in clinical strains,and to evaluate its clinical diagnostic and therapeutic value of gene.Methods The UL146 gene of clinical strains was examined by quantitative polymerase chain reaction（Q-PCR）or general polymerase chain reaction（PCR）.Positive samples of PCR amplification were sequenced and analyzed.Results High variability of UL146 gene was found among 28 HCMV strains.According to phylogenetic analysis,all sequences of UL146 in clinical strains could be divided into three types and four subtypes.Chemokine ELRCXC region was highly conserved in all sequences.Conclusion HCMV-UL146 genes showed a high degree of polymorphism,and its encoded chemokine ELRCXC region was highly con-served.The relationship between HCMV-UL146 gene′s polymorphism and different clinical symptoms of HCMV infection was unclear.

Objective To investigate the safety and efficacy of decitabine combined with CAG regimen in treatment of acute myeloid leukemia (AML) ineligible for conventional chemotherapy. Methods The data of 20 cases with AML ineligible for conventional chemotherapy from January 2013 to May 2015 were retrospectively analyzed. Decitabine combined with CAG regimen was used during induction therapy. The primary induction regimen was used 26 times after remission, the standard 3+7 regimen were used 7 times, and intermediate-dose cytarabine were used 3 times. The total course of treatment included 2-8 cycles. Results All of the 20 patients completed the first cycle of induction therapy, including 11 cases of complete remission (CR), 5 cases of partial remission and no response in 4 cases, and the overall response rate (ORR) was 80 % (16/20). ORR was 69.2 % (9/13) and 100.0 % (7/7) in high-risk group and middle-low risk group respectively. ORR was 60.0%(6/10) in AML evolving from MDS. 8 patients were infected during the induction therapy and the infection rate was 40.0% (8/20). 2 patients were died of pulmonary infection. The median number of suspended red blood cell and platelet infused were (9.1±5.7) U and (57.5±51.9) U respectively. Neutrophil recovery time was (8.7±5.6) days during induction therapy. All patients were followed up for at least 1 year, and 12 cases were dead. Overall survival rate was 85.0%at 3 months, 80.0%at 6 months, and 40.0%at 1 year. While in 12 CR patients relapse-free survival rate was 75.0%at 3 months, 75.0%at 6 months,and 65.6%at 1 year respectively. Conclusion Decitabine combined with CAG regimen with high remission rate and well tolerance, can be used as a first therapy for AML ineligible for conventional chemotherapy.

OBJECTIVETo analyze the nature of the protein encoded by 8B7cDNA (1 835 bp) and to examine the localization of the protein in cells.

METHODSThe nature of the protein was analyzed using Blastn, Blastp, and TMpred. Expressions of 8B7 mRNA in tissues and cells were examined by Northern blotting. Recombinant expression vectors for localization test were constructed and transfected into COS-7 cells. Fluorescence emission in cells was examined upon a laser scan confocal microscope.

RESULTSThe protein encoded by 8B7cDNA was 363 amino acids long and contained spectrin repeats. It was completely homologous to the C-terminal 363 amino acids of Enaptin, Nasprin-1, Mynel, and Syne-1 proteins. It belonged to Spectrin super-family and was called 8B7 spectrin. Northern blotting revealed a 1.8 kb mRNA expression in human spleen and small intestine tissues. EGFP-8B7 fusion protein was observed to express on the nuclear membrane and in the cytoplasm in a reticular form in a localization assay on COS-7 cells. The position of fluorescence in COS-7 cells transfected with pEGFP-delta SR8B7 was similar to that in the cells transfected with pEGFP-8B7cDNA.

CONCLUSIONS8 B7 spectrin is a novel member of spectrin super-family. It localizes to the nuclear membrane and the cytoplasm in a reticular form in COS-7 cells. The localization is determined by the C-terminal KASH domain in 8B7 spectrin molecule.

Amino Acid Sequence ; Animals ; Blotting, Northern ; COS Cells ; Cercopithecus aethiops ; DNA, Complementary ; genetics ; Humans ; Plasmids ; genetics ; RNA, Messenger ; genetics ; Sequence Homology, Amino Acid ; Spectrin ; biosynthesis ; chemistry ; genetics ; Transfection

Objective: To explore the effect of herb-partitioned moxibustion (HPM) on tight junctions (TJs) of intestinal epithelial cells in Crohn disease (CD) mediated by tumor necrosis factor-α (TNF-α)-nuclear factor kappa B (NF-κB)-myosin-light- chain kinase (MLCK) pathway. Methods: Forty-eight male Sprague-Dawley rats were randomly divided into a normal control (NC) group, a model control (MC) group, an HPM group and a mesalazine (MESA) group, with 12 rats in each group. Trinitrobenzene sulfonic acid (TNBS) was administered to establish CD models. When the model was confirmed a success, the HPM group rats were treated with HPM at Tianshu (ST 25) and Qihai (CV 6), while the MESA group rats were given MESA solution by lavage. When the intervention finished, the colonic epithelial tissues were separated, purified and cultured in each group to establish the intestinal epithelial barrier model in vitro, and TNF-α was added (100 ng/mL) in the culture medium and maintained for 24 h to establish an increased epithelial permeability model. Transepithelial electrical resistance (TEER) was used to examine the permeability of the barrier; Western blot was used to observe the expressions of the proteins related to TJs of intestinal epithelial cells mediated by TNF-α-NF-κB-MLCK pathway; immunofluorescence staining was used to observe the expressions and distributions of tight junction proteins in the intestinal epithelium. Results: After TNF-α induction, compared with the MC+TNF-α group, the TEER value increased significantly in the HPM+TNF-α and MESA+TNF-α groups (both P<0.001); the expressions of nuclear factor kappa B (NF-κB) p65, MLCK, myosin light chain (MLC), tumor necrosis factor receptor-associated factor 6 (TRAF6) and receptor interaction protein-1 (RIP1) decreased significantly (P<0.01 or P<0.05), and the expression of zinc finger protein A20 (A20) increased significantly (P<0.01); the expressions of occludin, claudin-1, zonula occludens protein 1 (ZO-1) and F-actin also increased significantly (all P<0.01). Compared with the MESA+TNF-α group, the expressions of MLC, occludin, claudin-1, ZO-1 and F-actin increased significantly in the HPM+TNF-α group (P<0.01 or P<0.05). Conclusion: HPM can protect or repair the damage of intestinal epithelial barrier in CD rats, which may be achieved through modulating the abnormal TJs in intestinal epithelium mediated by TNF-α-NF-κB-MLCK pathway.

Objective - To establish a new non exposed intratracheal instillation method for establishing a rat silicosis model. Methods ， The specific pathogen free SD rats were randomly divided into control group and experimental group with ten rats in ， each group. Rats in the control group were given 1.0 mL of 0.9% sodium chloride solution and rats in the experimental group - were given 1.0 mL of silica suspension with a mass concentration of 50 g/L adopting to the one time intratracheal instillation ， - ， method and then followed by ventilator assisted ventilation immediately. When the tidal volume stabilized at 2.0 mL the ventilator was removed and the tracheal intubation was pulled out. Five rats in each group were sacrificed after two and four ， - Results weeks after modeling and hematoxylin eosin staining and Masson staining of lung tissue were performed. There was ， ， no death in the two groups of rats during the experiment. After two and four weeks the control group had normal lung structure ， ， ， normal alveolar cavity size no inflammatory cell infiltration thin alveolar wall only a small amount of collagen distribution ， around the lung interstitium and bronchus. At the second week of modeling the alveolar wall of the rats in the experimental ， ， ， group was slightly thickened interstitial lymphocytes and macrophages were infiltrated slight hyperplasia was found and a ， small amount of fibroblasts were visible. At the 4th week of modeling the alveolar wall of the rats in the experimental group was ， ， ， ， significantly thickened fibrous nodules were formed and fibroblasts fibrocytes collagen fibers were significantly increased. Conclusion - The combination of ventilator and non exposed intratracheal instillation method can be used to successfully ， ， . establish a rat silicosis model which is simple safe and effective