OBJECTIVE To construct a lentiviral vector of RNA interference (RNAi) of MMP-9 gene and observe its inhibitive role on the invasion of laryngeal cancer cells.METHODS The effective sequence of siRNA targeting MMP-9 gene was confirmed.Both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pLVTHM vector,which contained H1 promoter and green fluorescent protein (GFP).The resulting lentiviral vector containing MMP-9 shRNA was called LV2 shMMP-9,and it was confirmed by PCR and sequencing.After that,MMP-9 shRNA was transfected into Hep-2 cells and western blot was used to test the expression of MMP-9.At last,Boyden Chamber was used to observe the invasion of the Hep-2 cells. RESULTS PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of MMP-9 (LV2shMMP-9) producing MMP-9 shRNA was constructed successfully. The titer of concentrated virus was 8?1010TU /L. Western blot showed that the expression of MMP-9 was negative in the MMP-9 siRNA transfected Hep-2 cells. And Boyden Chamber showed the invasive capability of Hep-2 cells transfected MMP-9 siRNA were obviously decreased.CONCLUSION The lentivirus RNAi vector of MMP-9 was constructed successfully,and MMP-9 silence can inhibit invasion of laryngeal cancer in vitro.

Objective To investigate the pathogenesis of avascular necrosis of the femoral head(ANFH) and to observe the effects of uhrashortwave diathermy treatment of ANFH through animal experiments.Methods A total of 30 New Zealand white rabbits were randomly divided into three groups:a control group,a model group,and a diathermy group.All the groups were injected with horse serum and methylprednisolone to induce ANFH.The path- ological effects were observed.Results The amount of osteoblast in the model group was significantly less than in the control group,while in the diathermy group it was significantly increased compared with the control group.The a- mount of osteoclast in the model and diathermy groups was significantly higher than in the control group,and in the diathermy group it was significantly more than in the model group.The thickness of femoral head cartilage in the mo- del and diathermy groups was reduced compared with the control group,while it was thicker in the model group than in the diathermy group.The empty cartilage cell lacunae ratios of the model and diathermy groups were significantly higher than for the control group,and the diathermy group showed significant degradation compared to the model group.The density of blood vessels under the cartilage in the model group was significantly less compared with the control group,while in the diathermy group it was significantly increased compared with the control group.The width of bone trabeculae in the model and diathermy groups was significantly less compared with the control group,while they were significantly wider in the diathermy group compared with the model group.The diameters of fat cells in the model and diathermy groups were increased compared with the control group,while they were significantly smaller in the dia- thermy group compared with the model group.The adipocyte area rates in the model and diathermy groups were signifi- cantly elevated compared with the control group,and rates in the model group were significantly elevated compared with the diathermy group.Conclusion Ultrashortwave diathermy is an effective treatment for early stage ANFH.

The microstructure of cationic cyclopeptide (TD-34) treated Caco-2 cell membrane was observed, and we discussed the relationship between membrane structure and insulin transmembrane permeability. Atomic force microscope (AFM) was used to observe living cell membrane in air condition and tapping mode. Results showed that the surface of Caco-2 cell membrane treated with TD-34 lost its smoothness and nearly doubled its roughness. Apparent permeability coefficients (P(app)) of insulin in Caco-2 cell monolayers increased 2.5 times. In conclusion, AFM can be used to observe microstructure of cationic cyclopeptide treated cell membrane and cationic cyclopeptide enhanced insulin delivery across Caco-2 cell membrane by increasing membrane fluidity.
Caco-2 Cells ; Cations ; Cell Membrane ; drug effects ; Cell Membrane Permeability ; drug effects ; Humans ; Insulin ; metabolism ; Membrane Fluidity ; drug effects ; Microscopy, Atomic Force ; Peptides, Cyclic ; pharmacology

Objective To study the effects of citrullinated vimentin (cVim) on the maturation and immunologic function of dendritic cells (DCs) from rheumatoid arthritis (RA) peripheral blood.Methods In the present study,mononuclear cells were isolated from the peripheral blood of patients with RA and cultivated in media containing GM-CSF and IL-4 to generate immature DCs (imDCs).The imDCs generated were stimulated with citrullinated vimentin and vimentin.LPS was used as the positive control and PBS was used as the negative control.The expression of surface molecules on the DCs,such as CD14,CD80,CD83,CD86,MHC Ⅰ and MHC Ⅱ were analyzed with FACS.The capability of the stimulatory activity of the DCs on allogeneic T cells in mixed reaction was tested by MTS.t-test was used for statistical analysis.Results Compared to untreated DCs,DCs treated with LPS increased the expression levels of MHC Ⅱ,CD80,CD83 and CD86 (1.07±0.14,1.25±0.13,1.90±1.08,2.44±0.65,P＜0.05),while cVim increased the expression levels of MHC Ⅱ ( 1.18±0.09,P＜0.05) and CD83 ( 1.97±0.99,P＜0.01 ),and Vim decreased the expression levels of CD80 (0.82±0.18,P＜0.01 ).It was demonstrated that the expression levels of MHC Ⅱ on DCs pulsed with cVim were significantly higher than that of the DCs with LPS,but the expression levels of CD80 and CD86 were not significantly different.The expression levels of MHC Ⅱ and CD83 on DCs pulsed with cVim were significantly higher than that of the DCs with Vim.The mixed lymphocyte reaction showed that the DCs induced by LPS and cVim trigerred the proli-feration of allogenic T cells obviously.Conclusion This result suggests that cVim could promote the phenotypic maturation of DCs and increase the expression of costimulatory molecules.

Adolescent ; Adult ; Female ; Graft vs Host Disease ; etiology ; prevention & control ; HLA Antigens ; immunology ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Histocompatibility Testing ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; therapy ; Survival Analysis ; Young Adult

Objective To investigate the effects of the HLA-Ⅰ and co-stimulators CD_(80)and CD_(86)on HT-29 cells surface, after heat killed Bifidobacterium and HT-29 cells were mingled culturing for 12 hours. Methods The HLA-Ⅰ and co-stimulators CD_(80)and CD_(86)on HT-29 cells surface were measured by Flow Cytometer. Results Heat killed Bifidobacterium can improve the expressions of HLA-1, CD_(80)and CD_(86)on HT-29 cells significantly, the livability of colon carcinoma cell was more than 90 %. Conclusion Heat killed Bifidobacterium have potential antitumorous features.

Objective To explore the effects of hyperglycemia and oxidized low density lipoprotein (ox-LDL) on the differentiation of macrophage derived THP-1 monocytes. Methods THP-1 human monocytic leukemia cell line was cultured in vitro, and the differentiation of THP-1 cells into macrophages was induced by phorbol esters. The macrophages were then incubated with the absence of D-glucose and ox-LDL (control group), 30 mmol/L D-glucose (hyperglycemia group), 100 μg/mL ox-LDL (ox-LDL group) or 30 mmol/L D-glucose and 100 μg/mL ox-LDL(G-ox-LDL group) for 24 h. High performance liquid chromatography was used for qualitative and quantitative analysis of intracellular cholesterol and cholesteryl esters. Both light microscope with red oil O staining technique and transmission electron microscope were employed to observe the morphology of treated and control THP-1 cells. Results A large number of intracellular red oil O stained granules and lipid vacuoles were observed in ox-LDL group and G-ox-LDL group, the contents of total cholesterol and cholesteryl esters were significantly higher than those of control group (P<0.05), and the contents of cholesteryl esters were higher than 50% of total cholesterol in both groups. However, only a few intracellular red oil O stained granules and lipid vacuoles were observed in control group and hyperglycemia group, there was no significant difference in the contents of total cholesterol and choleateryl esters between control group and hyperglycemia group (P>0.05), and the contents of cholesteryl esters were less than 50% of total cholesterol in both groups. Conclusion Foam cells form when THP-1 cells are incubated with ox-LDL, while hyperglycemia alone can not convert THP-1 cells to foam cells, indicating that ox-LDL is necessary for the macrophages derived THP-1 monocytes to turn into foam cells.

Objective To analyze the curative effects and complications of ventriculo-peritoneal shunt in treating hydrocephalus caused by different diseases in children. Methods The data of 289 children with hydrocephalus who were performed ventriculo-peritoneal shunt were examined and the incidence rate of complications was analyzed. Results Thirty-seven incidences of complications occurred in 24 patients with the total incidence rate of 12.46%.There was obstruction of proximate tube in five cases,distal end in two cases,with the obstruction rate of 2.42%.Postoperative infection happened in nine cases(3.11%).Bacterial culture was positive in three cases,two of which were Staphylococcus epidermidis and Bacillus coli in the other one.Negative results were found in six patients for the bacterial culture.Subdural hematoma occured in one case,and tube break after injury in two cases.Slit ventricle syndrome broke out seven times in three cases,leakage of cerebrospinal fluid in two,exposure of pumps in three,and subcutaneous hydrops in two.Four cases(1.38%)were dead from infection. Conclusion Ventriculo-peritoneal shunt is preferred for hydrocephalus in children if the complications are well prevented and controlled.

Objective To compare the thickness of the retina and its sublayers between young and elderly rats using a stereological method. Methods Six young(3 months old) and six elderly(2 year old) LongEvans rats were used in this study.The right eyeball was dissected from each rat and prepared as a set of serial sagittal sections and applied with HE staining.The sections and fields were sampled in the systematic random fashion and examined under a light microscope.The thickness of the whole retina and its 8 sublayers were identified and measured. Results Compared with that of the young rats,the thickness of the whole retina and most of the sublayers of the elderly rats were significantly decreased.The decrease was such so that the proportion of the thickness of each sublayer to that of the whole retina remained unchanged.Most interestingly,among the 8 sublayers of the retina,the thickness of the exterior plexus layer reduced nearly 46.2% during aging process.Conclusion Aging has a significant effect on the thickness of the rat retina.Such effect is better presented with the systematic random sampling method.

Objective To investigate the relationships between hyperuricaenia,serum uric acid (SUA) level and the chronic kidney disease (CKD) in adult residents of Pudong New Area,Shanghai.Methods 3326 residents aged 20-80 years were randomly selected from Pudong New Area,Shanghai through multistage sampling and interviewed between April and July of 2008.Fasting blood sample and morning ovid urine sample were collected for each participant for testing of SUA,serum creatinine,urinary albumin and creatinine.Both urine albumin to creatinine ratio (ACR)and glomerular filtration rate (GFR) were calculated to estimate the renal function.Results The overall prevalence of CKD was 16.0％ (age standardized 13.2％ ).The mean values of estimated GFR in participants with CKD and without CKD were (89.19 ± 27.25) and ( 105.88 ± 98.37) ml· min-1 ·(1.73 m2) -1,respectively.The prevalence rates of CKD in serum uric acid quartiles:first quartile,less than 4.2 mg/dl; second quartile,4.2-5.0 mg/dl; third quartile,5.0-6.0 mg/dl; and fourth quartile,6.0 mg/dl or more were 13.9％,15.0％,15.8％and 19.4％ (P＜0.05) respectively,increasing along with the increase of SUA among both sexes.Compared to the serum uric acid first quartile,the multivariate-adjusted odds for CKD of the second,third and fourth quartiles were 1.19 [95％ confidence interval (CI):0.90-1.58],1.27 (95％ CI:1.02-1.70),1.28 (95％ CI:1.10-1.68),respectively. Conclusion Hyperuricaemia was independently associated with the increased prevalence of CKD among population living in the Pudong New Area,Shanghai.