Aim To investigate the therapeutic effects of Iptakalim hydrochloride on brain damage and blood rheological changes induced by focal cerebral ischemia. Methods The rat model of focal cerebral ischemia was induced by middle cerebral artery occlusion using a suture method. Infarct size was evaluated by TTC staining. Neurological deficit score was evaluated by Longas method. Cerebral water content of the ischemic hemisphere was measured using wet-and-dry-weight method. The erythrocyte membrane fluidity was studied by the fluorescence polarization technique with DPH fluorescence probe. Results Infarct size of the brain was (24.75?6.66)% of contralateral hemisphere 6 h after focal cerebral ischemia.Pretreatment of Ipt 1.0～4.0 mg?kg -1,given inperitoneal,decreased the infarct size by 0.95%,6.6%(P

Objective To detect the atherosclerotic progression in low density lipoprotein receptor (LDL-R) gene knock-out mouse by ultrasound biomicroscopy(UBM) technique,and to monitor the intima media thickness (IMT) and changes in plague of aortic wall.Methods 10 male LDL-R gene knock-out mice of 16 weeks age and 10 LDL-R gene knock-out mice of 24 weeks age were in the experimental group,and 10 male C57BL/6 mice of 16 and 24 weeks age were in the control group.The shapes of their aortic roots,ascending aorta,aortic arch and CCA were detected by UBM,and the IMT at aortic root view and carotid artery bifurcation were measured,then the data were compared with histopathology of the corresponding vascular segments.Results The difference between the IMT of aortic root and carotid artery bifurcation of the 16 week old LDL-R mice and the control group of the same age had no statistical significance.The difference between the IMT of carotid artery bifurcation of the 24-week-old LDL R mice and the control group of the same age had no statistical significance.The IMT of aortic root thickened compared with control group of the same age(P ＜0.01).Conclusions The UBM technique can be used to detect the atherosclerotic progression in LDL-R gene knock-out mouse.

Objective To discuss the effect of 5-Aza-CdR on pancreatic carcinoma Capan-2 cell proliferation and PCDH8 gene expression.Methods Pancreatic carcinoma Capan-2 cells were treated with different doses of 5-Aza-CdR with or without gemcitabine,negative control group without drug,0.08μmol/L group with 0.08μmol/L 5-Aza-CdR,0.40 μmol/L group with 0.40 μmol/L 5-Aza-CdR,2.00 μmol/L group with 2.00 μmol/L 5-Aza-CdR,10 μmol/L group with 10 μmol/L 5-Aza-CdR,50 μmol/L group with 50 μmol/L 5-Aza-CdR. The inhibition ratio of Capan-2 cell proliferation were observed by MTT assay and PCDH8 gene and protein expression were detected by RT-PCR and Western blot. Results The inhibition ratio was increased with 5-Aza-CdR dose increasing(P<0.01),but decreased apparently with times extending(P<0.01). Inhibition ratio in 5-Aza-CdR and gemcitabine group was higher than those with only 5-Aza-CdR or gemcitabine groups(P<0.05 or P<0.01).The levels of PCDH8 gene and protein expression were increased significantly in 5-Aza-CdR treatment groups,with dose dependent (P <0.01 ).Conclusion 5-Aza-CdR can inhibit the proliferation of pancreatic carcinoma Capan-2 cell proliferation,and increase PCDH8 gene expression. The inhibition effect is strong when combined with gemcitabine.

Objective To investigate therapy effect and its clinical complications of the clavicular hook plate internal fixation for the treatment of Type Neer Ⅱ distal clavicle fracture and tossy Ⅲ acromioclavicular joint dislocation.Methods Twenty-two patients of Neer Ⅱ distal clavicle fracture and 16 patients of Tossy Ⅲ acromioclavicular joint dislocation were selected as our subjects.All patients were hospitalized and treated by using clavicular hook plate fixation form August 2007 to Februay 2012.The clinical effect and complications were analyzed retrospectively.Results All patients were showed the primary healing in terms of incision.Of all patients,30 cases were being following up,and follow-up periods was 6.0 to 18.0 months (average of (10.45 ±3.78) months).All patients underwent implant removal surgery at postoperative 6-14 months.No refracture or acromioclavicular joint separation again occurred.Four patients were with complications,of which,2 patients were clavicular hook plate decoupling at 7 days or 3 weeks after operation; One patient occurred clavicle stress fracture at more than 3 months after operation.All mentioned 3 cases were treated with re-operation.1 patient was with shoulder abnormal sound after operation and abnormal sound disappear after internal fixation removed.The shoulder functions were evaluated according to University of California-Los Angeles (UCLA) score system at before and after clavicular hook plate was taken out.The scores of pain and function of the shoulder,forward lateral flexion range were significant difference between before and after clavicular hook plate removed ((7.90 ±1.20) vs.(9.20±1.03),t =-2.327,P=0.045;(8.00±0.94) vs.(9.40 ±0.97),t=-3.280,P=0.001 ; (4.00 ±0.47) vs.(4.70 ± 0.48),t =-4.583,P =0.001).After the hook plate was removed,all shoulder functions showed improvement.The overall efficacy was excellent in 15 cases,good in 13 cases,and poor in 2 cases,and the recovery of excellent and good rate were 93.33％.Conclusion The clinical efficacy is well in terms of using clavicular hook plate internal fixation to treat Type Neer Ⅱ distal clavicle fracture and Tossy Ⅲ acromioclavicular joint dislocation.Prevention and treatment of complications should paid more attention.

Objective To investigate the effect of HBV on the expression of fibrosis-related factors in hepatic stellate cells(HSC)and its relation with liver fibrosis.Methods HSCs were co-cultured with HepG2 or HepG2.2.15 in vitro and HSCs cultured alone served as the control.The mRNA expression of matrix metalloproteinase(MMP)-2 and tissue inhibitor of metalloproteinase(TIMP)-1 was detected by realtime PCR.The protein expression of MMP-2 and TIMp-1 was detected by Western-blot.Results Compared with the control and the HSCs co-cultured with HepG2,the expression of MMP-2 and TIMP-1 mRNA in HSCs co-cultured with HepG2.2.15 was increased remarkably and the most significant difference was found at 72 h(F=11.91,23.13;P=0.008,0.001);the expression of MMP-2 and TIMP-1 protein in HSCs co-cuhured with HepG2.2.15 was also increased remarkably and the most significant difference was found at 72 h(F=20.70,6.54;P=0.002,0.003)too.Conclusion The expression of fibrosis-related factors in HSCs increased significantly after co-cultured with HepG2.2.15,which suggests that HBV could promote liver fibrosis.

BACKGROUND:Activation of Notch signaling plays a critical role in stem cel differentiation, and this effect seems to be cel-type dependent. Little is reported on the role of activation of Notch1 signaling in the differentiation of c-Kit+ bone marrow mesenchymal stem cels. OBJECTIVE:To analyze the influence of activation of Notch1 signaling on the differentiation of c-Kit+ bone marrow mesenchymal stem cels. METHODS:The Notch1 intracelular domain (N1-ICD) was obtained from the cDNA library by PCR and cloned intoBamHI/AgeI digested adenoviral GV314 plasmid to construct N1-ICD overexpressing shuttle plasmid, and the positive clones were verified by sequencing. N1-ICD shuttle plasmid and helper plasmids pBHGloxΔE1,3 Cre were used to co-transfect HEK293T cels to obtain N1-ICD overexpressing adenoviral particles (N1-ICD-Ad). The c-Kit+ subpopulation were isolated from bone marrow mesenchymal stem cels of the Sprague-Dawley rat femurviamagnetic activated cel sorting. After transfection of the c-Kit+ BMSCs with N1-ICD-Ad adenovirus, we assessed the activation of Notch1 signaling and differentiation in c-Kit+ bone marrow mesenchymal stem cels by quantitative RT-PCR and immunofluorescent staining. RESULTS AND CONCLUSION:N1-ICD coding sequence was successfuly generated from the cDNA library, and then was cloned into the linearized adenoviral vectors GV314. The resistant clones were verified by sequencing. With the assistance of packaging plasmids, recombinant N1-ICD-Ad adenovirus plasmids were successful packaged in HEK293T cels, and its title was 2×1012 PFU/L. c-Kit+ bone marrow mesenchymal stem cels with the purity of 91.6% were successfuly isolated from the bone marrow mesenchymal stem cels of the Sprague-Dawley rat femur. Compared with the blank and negative controls, N1-ICD-Ad infection in the c-Kit+ bone marrow mesenchymal stem cels led to substantial accumulation of N1-ICD in the cytoplasm and nuclei, significantly unregulated expressions of Hes1 (a downstream gene of Notch) and cardiomyocyte differentiation genes Nkx2.5 and cTnT, significantly increased the expression of von Wilebrand factor, an endothelial cel differentiation gene, and mildly increased the expression of smooth muscle22α, a smooth muscle cel differentiation gene. These experimental results indicate that the activation of Notch1 signaling contributes to multi-lineages differentiation of c-Kit+ bone marrow mesenchymal stem cels, and the construction of N1-ICD overexpressing adenoviral vector makes the foundation for further research on the role of Notch1 signaling in stem cel biology.

OBJECTIVE: To observe the clinical efficacy of highly agglutinative staphylococcin (HAS) combined with cisplatin in the treatment of malignant hydrothorax after straining and inserting central venous catheter in thoracic cavity. METHODS: 90 patients with malignant hydrothorax combined with lung cancer were randomly divided into A、B、C 3 groups. Central venous catheter was inserted in thoracic cavity for strainage. Group A were intrapleural injected with 5 000 U HAS, group B 40 mg?m-2 cisplatin and group C 5 000 U HAS combined with 40 mg?m-2 cisplatin once a week. Clinical efficacies of 3 groups were evaluated after 3 weeks of treatment. RESULTS: The total effective rate of group A, B and C were 43.0%, 46.7% and 80.0% respectively. There was obvious difference between group C and A as well as group B(P

Objective To investigate the protective and cure effects of dexamethasone on bleomycin-induced pulmonary fibrosis.Methods32 rats were randomly divided into the control group (C-group, n=8), injury group (I-group, n=12) and dexamethasone group (D-group, n=12). The acute pulmonary model was established by intratracheal injection of bleomycin with rats of the I-group and D-group; while rats of the C-group injected with distilled water. After that, rats of the D-group were injected with dexamethasone sodium phosphate in intraperitoneal every day, those of the C-group and I-group were injected with saline. The animals were killed on the 3rd, 7th, 14th, and 27th days after treatment, and tests of bronchoalveolar lavage fluid (BALF), total lung collagen content and lung tissue processing were performed.ResultsPathological evidence of the I-group rats demonstrated that the alveolar compartment companied with massive inflammatory cell invasion and a number of myofibroblast proliferation became more thick. However, lung injury in the D-group rats got better than that in the I-group. Neutrophil percentage achieved peak in both I-group and D-group on the 7th day. But the neutrophil ratio in the D-group was significantly lower than that of the I-group on the 7th day ( P<0.05) and the 14th day ( P<0.01). Total lung collagen content achieved peak on the 14th day both in I-group and D-group, but that of the I-group was significantly higher than that of the C-group ( P<0.01) and the D-group was significantly lower than the I-group ( P<0.01).ConclusionDexamethasone plays a protective and cure role in lung fibrosis by efficiently inhibiting the gather and invasion of neutropils and restraining the increase of collagen secreted by proliferous fibroblasts.

Humans ; Genetic Markers ; Family ; DNA Fingerprinting ; Microsatellite Repeats/genetics*

AIM: To explore the expression and significance of receptor tyrosine kinase anexelekto (Axl) in nasopharyngeal carcinoma (NPC).METHODS: Immunohistochemistry was used to detect the Axl protein expression of 78 patients with NPC and 32 patients with nasopharyngeal chronic inflammation (NPI).The correlations between the Axl protein levels and the clinical parameters of NPC patients were analyzed.NPC cells were cultured in vitro, and the expression of Axl in well differentiated CNE1 cells, poorly-differentiated CNE2Z cells and undifferentiated C666-1 cells was detected by immunofluorescence staining.After treatment of the CNE1and C666-1 cells with Axl specific inhibitor TP-0903, CCK-8 assay was used to detect cell viability, flow cytometry was adopted to analyze the cell cycle distribution, qPCR was used to examine the mRNA levels of Axl and proliferating cell nuclear antigen (PCNA), and Western blot was used to examine the protein expression of Axl and p-Axl.RESULTS: Axl protein was localized in the cell membrane and cytoplasm.The rate of high expression of Axl in NPC was significantly higher than that in NPI (P<0.01).High Axl expression showed no correlations with NPC patients'' age, gender and M stage, while positively correlated with the clinical stage, T stage and N stage (P<0.05).Axl protein showed a low level in the CNE1 cells, but showed a high level in CNE2Z and C666-1 cells.TP-0903 inhibited cell viability in concentration and time dependent manners.TP-0903 at 2 nmol/L showed significant inhibitory effects, as evidenced by arresting the cell cycle at G0 phase and reducing Axl activity and PCNA expression.CONCLUSION: High expression of Axl promotes the clinical progress of NPC.TP-0903 significantly inhibits the viability of NPC cells, suggesting that Axl may be a valuable target in the NPC treatment.