To observe the expression of N - cadherin and fibronectin in retinal pigment epithelium ( RPE) cells in vitro under high glucose conditions, furthermore, to explore the effects of high glucose on epithelial -mesenchymal transition (EMT) in RPE cells.
●METHODS: Human RPE (hRPE) cells were cultured in vitro. Containing a final concentration of 60mmol/ L glucose was used for high glucose treatment. The cells were divided into normal glucose group (5. 5mmol/ L, NG) and high glucose group (24, 48 and 72h) respectively. The expression of N - cadherin and fibronectin in hRPE cells were evaluated by immunofluorescence and real -time PCR.
●RESULTS:RPE cells became disorganized and swollen over time under high glucose conditions, especially in 72h subgroup. lmmunohistochemical analysis revealed that the expression of N - cadherin in RPE cells under high glucose conditions was decreased compared with that in the control group, while the expression of fibronectin was increased. Real - time PCR results showed that the expression of N - cadherin mRNA in high glucose group was decreased at 24h compared with that in the control group, and declined markedly at 72h ( F = 12. 252, P =0. 000). There were no significant differences between the control group and the high glucose group at 24h, while the differences between the control group and the high glucose group (48 and 72h) were significant respectively (P < 0. 05 ). Meanwhile, the expression of fibronectin mRNA in RPE cells was increased in high glucose group at 24h, and reached the peak at 72h (F = 50. 543, P = 0. 000). There were no significant differences between the control group and the high glucose group at 24h. Compared with the control group, the expression of fibronectin mRNA in hRPE cells was increased significantly in high glucose group (48 and 72h) respectively (P= 0. 000, P= 0. 000).
●CONCLUSlON: The expression of epithelium marker N-cadherin is down - regulated under high glucose conditions in hRPE cells in vitro. Meanwhile, the expression of mesenchymal maker fibronectin is induced and appeared to EMT changes. Results of this study will enrich our growing understanding in proliferative diabetic retinopathy and hopefully lead to novel insights for the pathogenesis and therapeutic treatments.

OBJECTIVETo investigate the antiinflammatory activities of aqueous extract of Occimum gratissmium (OGE) with emphasis on expression of proinflammatory cytokines in Lipopolysaccharide (LPS)-stimulated epithelial cell BEAS-2B.

METHODSEffects of OGE on cell viability were determined by MTT assay. mRNA expression were analyzed by and reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR. Activation of kinase cascades was investigated by immunoblot. Intracellular reactive oxygen species (ROS) was analyzed by flow cytometry.

RESULTSOGE (<200 μg/mL) treatment or pretreatment and following LPS exposure slightly affected viability of BEAS-2B cells. Increase of interleukin (IL)-6 and IL-8 and the elevated level of intracellular ROS in LPS-stimulated BEAS-2B cells were diminished by OGE pretreatment in a dose-dependent manner. OGE suppressed inflammatory response-associated mitogen-activated protein kinases (MAPKs) and Akt activation. Additionally, OGE pretreatment increased level of cellular inhibitor of κBα (IκBα) and inhibited nuclear translocation of nuclear factor kappa B (NF-κB).

CONCLUSIONThese findings indicate that significant suppression of IL-6 and IL-8 expressions in LPS-stimulated BEAS-2B cells by OGE may be attributed to inhibiting activation of MAPKs and Akt and consequently suppressing nuclear translocation of NF-κB.

Cell Nucleus ; drug effects ; metabolism ; Cell Survival ; drug effects ; Cytosol ; drug effects ; metabolism ; Epithelial Cells ; drug effects ; enzymology ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; I-kappa B Proteins ; metabolism ; Interleukin-6 ; genetics ; metabolism ; Interleukin-8 ; genetics ; metabolism ; Intracellular Space ; drug effects ; metabolism ; Lipopolysaccharides ; pharmacology ; Mitogen-Activated Protein Kinases ; metabolism ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; Ocimum ; chemistry ; Phosphorylation ; drug effects ; Plant Extracts ; pharmacology ; Protein Transport ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reactive Oxygen Species ; metabolism ; Respiratory System ; cytology ; Water

AIM:To investigate the influence of signal transducer and activator of transcription 3(STAT3)on Warburg effect in the malignant transformation of WB-F344 rat hepatic oval cells.METHODS:The WB-F344 cells were treated with N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)and hydrogen peroxide(H2O2)to induce the malignant trans-formation.Evaluation of the transformed cells were measured by the soft agar colony formation assay and DNA aneuploidy with flow cytometry.The levels of glucose and lactate in the culture medium of the cells were detected by chromatography. The protein levels of alpha-fetoprotein(AFP),STAT3,p-STAT3 and glucose transporter 2(GLUT2)in the cells were ex-amined by Western blot analysis.The cell proliferation were evaluated by WST-1 assay,viable cell counting,measuring the S-phase fraction(SPF)and proliferation index(PI)using the data from flow cytometry analysis,and detecting proliferating cell nuclear antigen(PCNA)protein expression by Western blot.RESULTS:Compared with the control cells,the forma-tion of colonies in soft agar(P<0.05)and DNA aneuploidy(P<0.01)were elevated in transformed cells,and the ex-pression level of AFP was also augmented(P<0.05).The increases in the level of both glucose consumption(P<0.05) and lactate production(P<0.01)show that Warburg effect was enhanced in transformed cells.Meanwhile, the protein levels of GLUT2(P<0.01)and p-STAT3(P<0.01)in transformed cells were higher than those in the control cells.The cell proliferation parameters including SPF(P<0.01),PI(P<0.01), viable cell number and PCNA expression(P<0.01)in transformed cells were also elevated as compared with the control cells.Interestingly, stattic, an inhibitor of STAT3 activation,resulted in declines in glucose consumption(P<0.05)and lactate production(P<0.01)in the trans-formed cells.In addition,compared with transformed cells,formation of colonies in soft agar(P<0.01),DNA aneuploidy (P<0.01),AFP(P<0.05), GLUT2(P<0.05), and cell proliferation parameters including SPF(P<0.01), PI (P<0.01),viable cell number(P<0.05)and PCNA expression(P<0.05)were also decreased following stattic treat-ment in transformed cells.CONCLUSION:STAT3 promotes Warburg effect and cell proliferation probably by upregula-ting GLUT2 expression in the malignant transformation of hepatic oval cells.

Objective To investigate the macular vascular density after successful repair of rhegmatogenous retinal detachment (RRD) for one year using optical coherence tomography angiography (OCTA),and discuss the correlation between the macular vascular density and visual acuity.Methods Totally 42 patients of the RRD (42 eyes),their contralateral eyes (A group) and 42 patients of the normal eyes (B group) were recruited into this study.All participants underwent examination with best corrected visual acuity (BCVA) and OCTA.The difference in macular vascular density was compared and the correlation between BCVA and the vascular density was analyzed.Results The macular vascular density of superficial layer,deep layer and choroidal capillary layer was 0.422 4 ±0.089 3,0.4836 ±0.0748,0.527 1 ±0.039 0 in RRD group,respectively,0.469 3 ±0.112 5,0.550 0 ±0.074 0,0.546 2 ±0.034 3 in A group,respectively,0.5619 ±0.053 7,0.611 2 ±0.035 2,0.562 6 ±0.030 4 in B group,respectively.The macular vascular density was significantly decreased in RRD group when compared with A and B groups (all P ＜ 0.05).There was a positive correlation between BCVA and the macular vascular density in the deep layer and choroidal capillaries layer (r =0.629,0.654,both P =0.000).However,there's no correlation between the macular vascular density of superficial layer and BCVA (P =0.103).Conclusion All the macular vascular densities are decreased in patients of RRD after successful repair of retinal detachment one year later,which indicated that the blood flow does not completely recover.And there is a positive correlation between BCVA and macular vascular densities in deep layer and choroidal capillaries layer.And meanwhile,OCTA can objectively and effectively quantify the status of macular region blood flow.

AIM: To study the reliability of Pentacam, OCT, Tomey OA-2000, IOL Master700, and A ultrasound pachymetry in the measurement of central corneal thickness(CCT)in myopic eyes.

METHODS: CCTs of 56 myopic patients(112 eyes)were measured using Pentacam, RTVue OCT, Tomey OA-2000, IOL Master 700, and A ultrasound pachymetry(NIDEK US-500). The difference, correlation, and consistency were statistically analyzed using average values taken from the five devices and the measured values produced by each device.

RESULTS: CCTs of 112 eyes were determined using Pentacam(530.17±25.08)μm, OCT(519.79±26.90)μm, Tomey OA-2000(521.75±26.51)μm, IOL Master 700(519.53±28.15)μm, and A ultrasound pachymetry(542.23±26.88)μm. The average value across all five devices was 526.69±26.08μm. The results of root mean square error(RMSE)and theil inequality coefficient(TIC)showed that the degree of deviation from the average of the values measured using Pentacam was the smallest. The other four devices were, from least deviation to most, Tomey OA-2000, OCT, IOL Master 700, and A ultrasound pachymetry. Typical correlation analysis showed that the correlation coefficients between the measured values of Pentacam, OCT, Tomey OA-2000, IOL Master 700, and A ultrasound pachymetry and the average values were ρ=0.957, P=0.950, P=0.953, P=0.930, and ρ=0.949, respectively. The results of correlation coefficient showed that Pentacam and Tomey OA-2000 had the closest correlation with the average value. The other three were, in order of closest to furthest correlation, OCT, A ultrasound pachymetry and IOL master 700. The results also showed that the measurements of Pentacam and Tomey OA-2000 were more reliable, which was consistent with the clinical situation. The results of Bland-Altman analysis showed that the five groups of measurements were closely consistent with the average values, and the CCT measurements of Pentacam and Tomey OA-2000 were the most consistent with the average values.

CONCLUSION: Among the five instruments, Pentacam and Tomey OA-2000 produced results that were closest to the average value in CCT measurement, with the best correlation and consistency. Their measurement methods are more objective and safer than A-scan, so they may provide better reference data for clinical practice.

OBJECTIVETo obtain recombinant human interleukin 12 by genetic engineering, and to explore possibility of its clinical application in treatment of tumor and chronic hepatitis.

METHODSBicistronic expression vector P35-IRES-P40 was constructed for the simultaneous translation of IL-12 p35 and p40 cDNA subunit through internal ribosomal entry sites (IRES). pCI-dhfr-P35-IRES-P40 vector was constructed for expression in CHO-DHFR- cells. Positively cloned cells were screened by means of ELISA. Pools of clones with increased expression of IL-12 could be generated by selection in methotrexate. To determine the biological activities of rhIL-12, PHA-activated lymphoblasts proliferation assay and IFN-gamma induction assay were used in this study.

RESULTSGenetically engineered cells expressing hIl-12 were obtained and all the cell lines showed the stabile expression of rhIL-12 in high efficiency and good growth properties.

CONCLUSIONrhIL-12 have good biological activities, it can stimulate activation and proliferation of T cells and induce production of IFN-gamma.

Animals ; Blotting, Western ; CHO Cells ; Cell Proliferation ; drug effects ; Cricetinae ; Cricetulus ; Dose-Response Relationship, Drug ; Genetic Vectors ; genetics ; Humans ; Interleukin-12 ; biosynthesis ; genetics ; pharmacology ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; metabolism ; pharmacology ; T-Lymphocytes ; cytology ; drug effects ; Transfection

The acquisition of metastasis potential is a critical point for malignant tumors. Melanoma differentiation associated gene-7/interleukin-24 (mda-7/IL-24) is a potential tumor suppress gene and frequently down-regulated in malignant tumors. It has been implicated that overexpression of MDA-7 led to proliferation inhibition in many types of human tumor. Invasion is an important process which is potential to promote tumor metastasis. However, the role and potential molecular mechanism of mda-7/IL-24 to inhibit the invasion of human melanoma cancer is not fully clear. In this report, we identified a solid role for mda-7/IL-24 in invasion inhibition of human melanoma cancer LiBr cells, including decreasing of adhesion and invasion in vitro, blocking cell cycle, down-regulating the expression of ICAM-1, MMP-2/9, CDK1, the phosphorylation of ERK and Akt, NF-kappaB and AP-1 transcription activity. Meanwhile, there was an increased expression of PTEN in mda-7/IL-24 over-expression LiBr cells. Our results demonstrated that mda-7/IL-24 is a potential invasion suppress gene, which inhibits the invasion of LiBr cells by the down-regulation of ICAM-1, MMP-2/9, PTEN, and CDK1 expression. The molecular pathways involved were the MAPK/ERK, PI3K-Akt, NF-kappaB, and AP-1. These findings suggest that mda-7/IL-24 may be used as a possible therapeutic strategy for human melanoma cancer.
CDC2 Protein Kinase/genetics/metabolism ; Cell Line, Tumor ; Cell Movement ; Down-Regulation ; G2 Phase Cell Cycle Checkpoints ; Humans ; Intercellular Adhesion Molecule-1/genetics/metabolism ; Interleukins/genetics/*metabolism ; M Phase Cell Cycle Checkpoints ; Matrix Metalloproteinase 2/genetics/metabolism ; Matrix Metalloproteinase 9/genetics/metabolism ; Melanoma/metabolism/pathology ; NF-kappa B/genetics/metabolism ; PTEN Phosphohydrolase/genetics/metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt/genetics/metabolism ; Transcription Factor AP-1/genetics/metabolism ; Up-Regulation

OBJECTIVETo explore the feasibility of surgical design for mandibular retrusion using three-dimensional software.

METHODSThree-dimensional reconstruction was performed by Mimics software based on the preoperative CT data. The model of the maxillofacial region was imported into Rapidform software for measuring the associated parameters and Geomagic software for simulation of osteotomy. The reliability of the virtual operation was validated during the surgery.

RESULTSThe model of mandibular retrusion was reconstructed and successfully used to simulate the surgery. The simulation result was applied in subsequent actual surgery and good surgical outcomes were achieved.

CONCLUSIONThe three-dimensional software can be used to simulate the surgery for mandibular retrusion and improve the predictability and accuracy of the surgery.

Computer Simulation ; Feasibility Studies ; Humans ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; methods ; Male ; Malocclusion, Angle Class II ; surgery ; therapy ; Mandible ; abnormalities ; diagnostic imaging ; surgery ; Maxilla ; diagnostic imaging ; Models, Anatomic ; Retrognathia ; diagnostic imaging ; surgery ; Software ; Surgery, Computer-Assisted ; methods ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo study the epidemiological pattern and trends of hepatitis B virus (HBV) in the area where people had been immunized by HBV vaccine for long time.

METHODSThrough cluster sampling and cross-sectional study, relative information and blood samples from people in Long-an county by families were collected. Signals of HBV infection were tested by solid-phase reverse immunosorbent test.

RESULTS(1) The average HBsAg positive rate was 7.5% with anti-HBs as 44.5 %, and anti-HBc as 47.8%. The positive rates of HBsAg and anti-HBc among 0-19 year-olds were lower than those of > or = 20 year-olds. (2) The positive rates of HBsAg, anti-HBc and HBV infection among HBV vaccine immunized group were 2.8%, 12.0% and 12.5% respectively, comparing with which among the un-immunized group as 10.2%, 69.8% and 71.2% respectively. (3) The HBsAg positive rate of male was higher than the female's but with no significant difference of anti-HBs and anti-HBc between different sexes. (4) The average HBsAg positive rate of 0-19 years old group was only 2.4%, while that of 20-30 years old group was 13.6%-17.7% and dropped from 60 years old group and on. The anti-HBs positive rate of 0-19 years old people started to drop significantly by age. The anti-HBs and anti-HBc positive rates of > or = 20 years people were showing a rising trend by ages.

CONCLUSIONIt seemed obviously that the HBV epidemiological patterns had changed after HBV vaccine had been universally used for long time in newborns. The age peak of infection had been pushed backward for nearly 20 years. It had been proved that the HBV vaccine immunization program had obtained excellent efficacy.

Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Hepatitis B ; epidemiology ; immunology ; prevention & control ; Hepatitis B Antibodies ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B Vaccines ; therapeutic use ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Young Adult

OBJECTIVETo study the epidemic pattern and trend of HBV infection in the area where the people had been immunized by HBV vaccine for 20 years.

METHODSThe whole sampling method was applied in combination with cross-sectional investigation. Blood samples were taken from every member of families. Markers of HBV infection were determined by using solid-phase radioimmunoassay (SPRIA).

RESULTS(1) The average HBsAg positive rate was 7.5%. The positive rate of markers for HBV infection of 0-19 years old subjects were lower than those of > or = 20 years old subjects. (2) The positive rate of HBsAg of 0-19 years old subjects in 1985 was higher than that in 2005. The anti-HBs positive rate in 1985 stemmed to be higher with age. It was 12.4% in 1- age group to 53.8% in >60 years age group. While the result of 2005 showed that the anti-HBs positive rate of 0-19 years old subjects dropped with age. The anti-HBc positive rate in 1985 also tended to be higher with age. But the result of 2005 showed that the rate of 0-19 years old subjects was just 1.4% to 16.8%.

CONCLUSIONThe epidemic patterns of HBV infection have had significant variations in the target population. HBV vaccine immunization has obtained excellent efficacy.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Hepatitis B ; blood ; epidemiology ; prevention & control ; Hepatitis B Vaccines ; administration & dosage ; therapeutic use ; Humans ; Immunization ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Seroepidemiologic Studies ; Time Factors ; Treatment Outcome ; Young Adult