BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

Aim To investigate the effect of aloe-emodin on lipopolysaccharide(LPS)-induced production of nitric oxide and expression of inducible nitric oxide synthase in RAW264.7 cells.Methods RAW264.7 macrophage line in mice was induced by LPS to set up the inflammatory model.Nitric oxide(NO)production was examined by Griess reaction;the expression of iNOS mRNA was detected by RT-PCR analysis;NO radical generation was tested by sodium nitroprusside method.Results Aloe-emodin at the dose of 0.69～2.5 mg·L~(-1) exhibited the inhibitory effect on LPS-induced NO production in a dose-dependent and time-dependent manner;aloe-emodin at the dose of 0.63～5.00 mg·L~(-1) suppressed LPS-induced iNOS mRNA expression in RAW 264.7 cells.However,aloe-emodin had no scavenging effect on sodium nitroprusside-triggered NO production,and didn't affect iNOS enzyme activity.Conclusion Aloe-emodin inhibited signifi-cantly LPS-induced NO production through suppressing inducible NO synthase(iNOS)expression at mRNA level in a dose-dependent and time-dependent manner,but failed to affect sodium nitroprusside-triggered NO production and iNOS enzyme activity.

Objective To measure the in vitro antibacterial activity of tigecycline against carbapenems-resistant Acinetobacter calcoacetcus-Acinetobacter baumannii complex.Methods The isolated strains of carbapenems-resistant Acinetobacter calcoacetcus-Acinetobacter baumannii complex were collected in our hospital from December 2013 to February 2014.The MIC test strip was a-dopted to measure the MIC value of tigecycline.The break point adopted the judgment criteria published by FDA.Results All 61 strains of carbapenems-resistant Acinetobacter calcoacetcus-Acinetobacter baumannii complex had extremely high drug resistant rate to the commonly used antimicrobial agents.The sensitive rate of tigecycline was 80.3%,intermediation was 19.7% and no re-sistant strain was found in this study.MIC50 and MIC90 were 2 μg/mL and 3 μg/mL respectively.Conclusion Tigecycline has bet-ter in vitro antibacterial activity to the carbapenems-resistant Acinetobacter calcoacetcus-Acinetobacter baumannii complex isolated in our hospital.

Integrins are a family of cell surface glycoproteins that contribute to a variety of biological functions, including cell growth, migration, proliferation and morphology. In addition, integrins also play the important roles in pathological process. Several viruses have been showed to use integrins as receptors or co-receptors to infect host cells.This article mainly reviews the progress on integrins and their roles in FMDV infection.

Receptors are primary determinant of viral tropism and disease pathogenesis.Heparan sulfates (HS)are ubiquitous,polyanionic carbohydrate chains linked to core proteins in cell membranes and ex- tracellular matrices of all eukaryotes.HS have also been demonstrated to function as receptors or co-receptors for a number of different viruses.To date,HS and four RGD-dependent integrins,?v?3,?v?6, ?v?1,and?v?8 have been reported to serve as receptors for Foot-and-mouth disease virus(FMDV).Different receptors may be used to interact with host cells during FMDV infection.Studies on the structure and function of receptors are very important for understanding the interaction between host cells and FMDV. Here,We mainly reviews the progress on the biological characteristics of HS and its roles in FMDV infection.

OBJECTIVETo explore morphological character and clinical significance of superior-posterior acetabular wall by anatomically measuring and quantitatively analyzing thickness of posterior acetabular wall, then provide a theoretical reference for clinical treatment of acetabular fracture.

METHODSFifteen adult formalin-preserved cadaveric pelvises (8 males and 7 females) were used for this investigation. Excess soft tissue was removed and the whole acetabular posterior walls were marked with "angle" sector method and the thickness was measured with caliper in different levels of the different split points. The measurement results were validated and analyzed statistically.

RESULTSAt 5 mm away from acetabular rim, the average thickness of superior-posterior acetablar wall fluctuated between (6.47±0.61) mm and (7.43±0.71) mm; the average thickness of inferior-posterior acetabuluar wall fluctuated between (5.62±0.51) mm and (6.33±0.61) mm; the average thickness of acetabular roof fluctuated between (7.71±0.74) mm and (8.27±0.99) mm. There was no statistical difference between average thickness of superior-posterior wall of acetabulum and inferior-posterior wall of acetabulum (P>0.05), but the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P<0.05). At 10 mm away from the acetabular rim, the average thickness of superior-posterior acetabular wall fluctuated between (8.81±0.67) mm and (13.35±0.89)mm; the average thickness of inferior-posterior acetabular wall fluctuated between (7.02±0.63) mm and (7.66±0.69) mm; the average thickness of acetabular roof fluctuated between (14.46±0.97) mm and (17.05±1.35) mm. Comparatively, the average thickness of superior-posterior acetabular wall was significantly larger than inferior-posterior wall of acetabulum (P<0.05), and the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P<0.01). At 15 mm away from the acetabular rim, the average thickness of superior-posterior acetabular wall fluctuated between (12.08±0.78) mm and (19.84±1.03) mm; the average thickness of inferior-posterior acetabular wall fluctuated between (10.17±0.76) mm and (11.12± 0.77) mm; the average thickness of acetabular roof fluctuated between (23.23±1.12) mm and (26.01±1.53) mm. Comparatively, the average thickness of superior-posterior wall of acetabulum was significantly larger than inferior-posterior acetabular wall (P<0.01), and the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P< 0.01).

CONCLUSIONThe thickness of entire acetabular posterior edge revealed an increasing tendency from inferior-posterior wall to the superior-posterior wall to acetabular roof. And this trend became more obvious with increasing distance away from acetabular rim. Therefore, the superior-posterior acetabular wall could not only maintain the stability of hip joint but also bear loading.

Acetabulum ; anatomy & histology ; injuries ; surgery ; Female ; Humans ; Male

OBJECTIVETo study the influence of the reference values for semen analysis proposed in the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen on the indication spectrum for intrauterine insemination (IUI).

METHODSWe retrospectively analyzed the clinical data of 111 cycles of IUI by the reference values for semen analysis in the 4th edition of the WHO Laboratory Manual (group A) and 84 cycles by the 5th edition (group B). We recorded and compared the percentages of various indications for IUI between the two groups.

RESULTSThe complications for IUI in groups A and B were as follows: asthenospermia (87.4% [97/111] vs 55.9% [47/84], P < 0.05), oligospermia (0 vs 0), teratospermia (51.4% [57/111] vs 35.7% [30/84]) , abnormal liquefaction (0.9% [1/111] vs O) , sexual dysfunction and genital malformation (0 vs 3.6% [3/84] , immune infertility (0.9% [ 1/111] vs O), and unexplained infertility (3.6% [4/111] vs 2. 4% [2/84 ] ). There were no significant differences between the two groups in the percentages of all the indications except that of asthenospermia.

CONCLUSIONThe reference values for semen analysis proposed in the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen have an evident influence on the indication spectrum for IUI by largely reducing the cases of IUI for male factors, prolonging the cycles of some patients, causing excessive diagnosis and treatment for females, and increasing their mental and economic burdens.

Adult ; Contraindications ; Female ; Humans ; Insemination, Artificial ; Male ; Pregnancy ; Reference Values ; Retrospective Studies ; Semen ; Semen Analysis ; World Health Organization

BACKGROUNDLaparoscopic splenectomy (LS) for massive splenomegaly is more technically challenging than for a normal-sized spleen. The purpose of this study was to determine the effect of operative experience on perioperative outcomes of LS for massive splenomegaly.

METHODSBetween January 2008 and December 2010, 36 consecutive patients who were diagnosed with massive splenomegaly underwent LS in our department. The perioperative outcomes were evaluated for evidence of a learning curve effect. Patients were divided into three groups (1, 2, and 3) of 12 consecutive patients, and outcomes of each group were compared.

RESULTSThe mean operative time decreased significantly from 252 minutes of Group 1 to 179 minutes of Group 3. The estimated blood loss and length of post-operative hospital stay showed a similar trend. No significant differences were found in the splenic length and weight, transfusion rate, or average amount of drainage. In this cohort, there were three cases with surgical complications and one conversion to open laparotomy.

CONCLUSIONSThe first 24 cases constitute the early stage of the learning curve for LS for massive splenomegaly. LS for massive splenomegaly is a technically challenging operation with a long learning curve, and strategies for developing training programs must address these challenges.

Adult ; Female ; Humans ; Laparoscopy ; education ; Learning Curve ; Male ; Middle Aged ; Operative Time ; Retrospective Studies ; Splenectomy ; adverse effects ; education ; Splenomegaly ; surgery

AIMTo evaluate the characteristics, the hypoglycemic efficacy and the pharmacokinetics of the insulin-liposomes double-coated by chitosan (CH) and chitosan-EDTA conjugates (CEC).

METHODSInsulin-liposomes were prepared by reversed-phase evaporation. The protection of insulin against peptic and tryptic digestion was studied with HPLC. The hypoglycemic effects of insulin-liposomes were investigated using the glucose oxidase method after oral administration to rats. Serum insulin concentration in rats were determined by radio-immunoassay, and were assessed by Pkanalyst computer program.

RESULTSThe insulin-liposomes double-coated by CH and CEC was shown to protect insulin against digestion of pepsin, trypsin and gastrointestinal contents. In glucose tolerance test in normal rats, as compared with phosphate buffer solution control group, the insulin-liposomes coated by CH and CEC could reduce the glucose-induced peak of hyperglycemia. The reduction of the insulin-liposomes double-coated by CH and CEC was superior to that of other insulin-liposomes. When administered intragastrically to normal rats, the insulin-liposomes coated by CH and CEC could reduce glycemia measured after an overnight fast. The hypoglycemic effect the insulin-liposomes double-coated by CH and CEC was superior to that of other insulin-liposomes, and the dosage of 50 mu x kg(-1) decreased by 45.98% of initial blood glucose level at 1 h. As compared with subcutaneous injection, the relative pharmacological bioavailability was 17.02% calculated by area under the curve of glucose level versus time profile after oral administration of the insulin-liposomes double-coated by CH and CEC to rats. The serum insulin concentration-time curves were found to best fit the one-compartment open model. As compared with subcutaneous injection, the relative bioavailability was 8.91% calculated by the area under the curve of serum insulin concentration versus time profile after oral administration of the insulin-liposomes double-coated by CH and CEC to rats.

CONCLUSIONThe stability and absorption of insulin-liposomes double-coated by CH and CEC was superior to that of the insulin-liposomes coated either by CH, or by CEC respectively.

Administration, Oral ; Animals ; Biological Availability ; Blood Glucose ; metabolism ; Chitosan ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Edetic Acid ; chemistry ; Hypoglycemic Agents ; administration & dosage ; pharmacokinetics ; pharmacology ; Insulin ; administration & dosage ; pharmacokinetics ; pharmacology ; Liposomes ; Male ; Nanotechnology ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Technology, Pharmaceutical ; methods

OBJECTIVETo explore the changes of surface antigen and function of rituximab on dendritic cells derived from patients with Primary immune thrombocytopenia (ITP) to further understand the effective mechanism of immunotherapy.

METHODSThe peripheral blood mononuclear cells (PBMCs) were isolated from remission patients with ITP before and after low-dose rituximab infusion, and the PMNCs were stimulated for 5 days by rhGM-CSF and rhlL-4 in 5% CO2 air at 37°C incubator. Then all of DCs were cultured with TNF-α for 48 hours. The morphology of DCs was monitored under inverted microscope daily, and the surface antigens of the DCs were analysed by flow cytometry, meanwhile the levels of IL-12p70 and TGF-β1 in supernatants were detected by ELISA, mix lymphocyte reaction was performed by MTT assay.

RESULTS(1) Rituximab-treated-DCs showed no obvious tree-like protruding compared with untreated-DCs. The former cells were small and most of nucleus were centric. (2) The expressions of HLA-DR, CD80, CD83 and CD86 on rituximab-treated-DCs \[56.37 ± 3.95)%, (36.41 ± 2.82)%, (30.45 ± 4.61)% and (41.98 ± 4.17)%, respectively\] were significantly lower than those untreated-DCs \[(73.71 ± 7.61)%, (55.14 ± 7.30)%, (80.91 ± 7.09)% and (59.03 ± 3.43)%, respectively\](all P < 0.05), the concentration of IL-12p70 was significantly lower, \[(66.87 ± 4.29)% vs (50.17 ± 14.52)%\], while that of TGF-β1 \[(9.70 ± 0.31)%\] higher than the untreated-DCs \[(2.70 ± 0.36)%\] (P < 0.05). (3) The abilities to activate T cells proliferation of rituximab-treated-DCs reduced compared with untreated-DCs.

CONCLUSIONThe surface antigen of ITP-DCs and the concentration of IL-12p70 reduced after the low-dose rituximab infusion. The abilities to activate T cells proliferation reduced while the concentration of TGF-β1 increased. Rituximab may achieve its therapeutic effect on ITP by downregulating the immunoreactivity of DCs.

Antibodies, Monoclonal, Murine-Derived ; administration & dosage ; therapeutic use ; Cell Proliferation ; Cells, Cultured ; Dendritic Cells ; cytology ; metabolism ; secretion ; Female ; Humans ; Interleukin-12 ; metabolism ; Lymphocyte Activation ; Male ; Rituximab ; T-Lymphocytes ; immunology ; Thrombocytopenia ; drug therapy ; immunology ; metabolism ; Transforming Growth Factor beta1 ; metabolism