Objective:To investigate the effect of Rg1 on the expression of apoptosis related genes of LEC induced by experimental oxidative injuries.Methods:All fresh transparent lenses of rabbit except those in the control group were bathed in a culture medium with 300?mol/L hydrogen peroxide(H2O2)to establish cataract models in vitro.Meanwhile Rg1 were added to the culture medium and incubated for 24 hours.The anterior capsules were used in the following study.The expressions of apoptosis related genes Bcl-2 and Bax protein were studied by immunohisto chemical techniques.Results:Bcl-2 and Bax protein were expressed in LEC of normal rabbit lens.Expression of Bcl-2 protein was much stronger than that of Bax protein.The expression of Bcl 2 protein was down-regulated and that of Bax protein was up-regulated in H2O2 group when compared with the control group,so the ratio of Bcl-2/Bax was lower.Rg1 up-regulated the expression of Bcl-2,and down-regulated that of Bax protein;the ratio of Bcl-2/Bax was higher.There were significant differences in the range of ratio changes compared to that of the taurine group.Conclusion:Rg1 can up-regulate the expression of Bcl-2,and down-regulate that of Bax protein,to increase the ratio of Bcl-2/Bax;while the expression of Bcl-2 protein was down-regulated and that of Bax protein was up-regulated in the H2O2 group.The molecular mechanisms of both H2O2 inducing apoptosis of LEC and natural anti-oxidants restraining apoptosis of LEC may be related to the expressions of apoptosis-related gens Bcl-2 and Bax.

Objective To describe radiological features of bronchopulmonary dysplasia(BPD) in premature infants,and improve the diagnostic ability of BPD.Methods Eleven Premature infants with BPD underwent series of chest radiographs,X-ray features of BPD were analysed.Results The radiographic findings frequently included diffuse opacities,linear-reticular opacities,sacculiform radiolucent areas and over-inflation of lung.There were 4 cases with two of the radiological abnormalties obove,diffuse opacities and linear-reticular opacities in 2,diffuse opacities and sacculiform radiolucent areas in one and diffuse opacities and over-inflation in one.There were two cases with three kinds of X-ray features,including diffuse opacities,linear-reticular opacities and sacculiform radiolucent areas.There was one case with all of the radiological features noted above.Conclusion The chest radiographs appearance were of characteristics in premature infants with bronchopulmonary dysplasia.

OBJECTIVE :To develop a RP-HPLC method for determination of minocycline concentration in human plasma.METHODS:Chromatographic separation has been achieved on C18 column with acetonitrile-H2O-TFA(15∶85∶0.1)as the mobile phase, and oxytetracycline as internal standard.The detection wavelength was 350nm.The minocycline was extracted from buffered plasma(pH=6.5)by ethyl- acetate, and quantified by the ratio of minocycline peak area to that of internal standard.RESULTS :The linear range of minocycline detection concentration was 0.05～8?g/ml(r=0.9 999).The minimum detection concentration was 0.02?g/ml with an average recovery of 101.89% .The inter and intra-day RSD were both less than 5%.CONCLUSION :The present method is simple, rapid and accurate for determination of minocycline in human plasma.

0.05) among these pharmacokinetic parameters. The relative bioavailability of the test preparation was(107.8?30.0)%. CONCLUSION: The test and the reference preparation are bioequivalent.

Objective To observe the inhibitory effect of Tirofiban on different shear-induced platelet aggregation, and to provide medication suggestions for the treatment of thrombosis in different hemodynamic environment. Methods Polydimethylsiloxane ( PDMS)-glass microchannel chips were fabricated by soft lithography. The whole blood of healthy volunteers anticoagulated with sodium citrate was collected and incubated with different concentrations of Tirofiban in vitro. The blood flowed through the straight microchannel or channel with 80% narrow for 150 seconds at the speed of 11 μL/ min and 52 μL/ min, respectively. The wall shear stress rates in straight channel at 11 μL/ min and 52 μL/ min were 300 s-1 and 1 500 s-1, respectively. The maximum wall shear rates in the channel with 80% occlusion at 11 μL/ min and 52 μL/ min were 1 600 s-1 and 7 500 s-1, respectively. The adhesion and aggregation images of fluorescent labeled platelets on glass surface were photographed with the microscope, and the fluorescent images were analyzed with Image J. The platelet surface coverage ratio was used as a quantitative index of platelet aggregation behavior, and the IC50 of Tirofiban for platelet inhibition was calculated under different shear rates. Flow cytometry was used to detect the platelet activation index (CD62P, PAC-1) in the whole blood at 52 μL/ min in channel with 80% occlusion. Results Tirofiban inhibited platelet aggregation in a dose-dependent manner, and the inhibitory effect was related to the shear rate. Under the shear rates of 11 μL/ min and 52 μL/ min, the aggregation was almost completely inhibited when the concentration in straight channel reached 100 nmol / L. When the concentration in channels with 80% occlusion reached 1 μmol / L, the aggregation was almost completely inhibited. IC50 values at 11 μL/ min and 52 μL/ min in straight channel were 2. 3 nmol / L and 0. 5 nmol / L, respectively. IC50 values at 11 μL/ min and 52 μL/ min in channels with 80% occlusion were 20. 73 nmol / L and 4. 5 nmol / L. Pathologically high shearforce induced an increase in platelet activation, which could be inhibited by Tirofiban. Conclusions Tirofiban can effectively inhibit shear-induced platelet aggregation, and different concentrations of Tirofiban should be given according to the thrombus formed in different shear force environment in clinic practice

Objective:To explore the mechanism by which SRY-related high mobility group-box 9 (SOX9) promotes the epithelial mesenchymal transition (EMT) of non-small cell lung cancer (NSCLC) A549 cells via the Wnt/β-catenin pathway. Methods: The human NSCLCA549 cell line was divided into three groups: OE-NC group, OE-SOX9 group and OE-SOX9+XAV-939 group. The cells in OESOX9 group were transfected with SOX9 pcDNA plasmid to up-regulate the expression level of SOX9; The cells in OE-SOX9+XAV939 group were transfected with SOX9 pcDNA plasmid while the β-catenin inhibitor XAV-939 (1.0 μmol/L) was added to the medium. qPCR was used to detect SOX9 mRNA levels; CCK-8 was used to examine the proliferation of A549 cells; Wound-healing assay and Transwell chamber assay were used to detect the migration and invasion ofA549 cells, respectively; and WB was used to detect protein expressions of SOX9, β-catenin, E-cadherin, γ-catenin, N-cadherin and vimentin. Results: The mRNA and protein levels of SOX9 in OE-SOX9 group and OE-SOX9+XAV-939 group were significantly higher than those in the OE-NC group after transfection (all P< 0.05), while there was no significant difference between the OE-SOX9 group and the OE-SOX9+XAV-939 group (P>0.05). The proliferation, migration and invasion of cells in OE-SOX9 group were significantly higher than those in OE-NC group; however, those abilities in OE-SOX9+XAV-939 group were significantly lower than those in OE-SOX9 group (all P<0.05). The level of β-catenin protein in OE-SOX9 group was significantly higher than that in the OE-NC group, while the level of β-catenin protein in OE-SOX9+XAV-939 group was lower than that in OE-SOX9 group (all P<0.05). Compared with the OE-NC group, the levels of phenotypic markers of epithelial cells, E-cadherin and γ-catenin, were down-regulated, and the phenotypic markers of mesenchymal cells, N-cadherin and vimentin, were up-regulated in cells of OE-SOX9 group; however, E-cadherin and γ-catenin were higher, and N-cadherin and vimentin were lower in OE-SOX9+XAV-939 group than those in OE-SOX9 group (all P<0.05). Conclusion: SOX9 could promote proliferation, migration and EMT of NSCLCA549 cells by activating the Wnt/β-catenin pathway. ··

Objective: To explore the protective effect and its molecular mechanism of apoptosis signal-regulating kinase 1 (ASK1) inhibitor (GS-459679) on acetaminophen-induced liver injury in mice. Methods: The model of liver injury was established by administration of acetaminophen (APAP) (300 mg/kg, i.p.) on C57BL/6 mice. Forty-eight male C57BL/6 mice were randomly divided into four groups, consisting of control group, GS group (GS-459679, 30 mg/kg, i.p.), APAP-induced group, and GS combined with APAP-induced group. For GS combined with APAP-induced group, mice were treated with GS 30 min prior to administration of APAP. After mice were euthanized at 6 h or 12 h, respectively, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed, and mRNA levels of TNF-α, IL-6 and IL-1β were tested. The activity of glutathione (GSH), oxidized GSH (GSSG) and malondialdehyde were quantified. In addition, ASK1, P-ASK1, JNK and P-JNK protein levels were tested in all groups. Results: The ASK1 and P-ASK1 levels were up-regulated in APAP-induced group. Compared to the control group, serum levels of ALT and AST, and mRNA levels of TNF-α, IL-6 and IL-1β were increased in APAP-induced group. Meanwhile, the levels of MAD and GSSG, and the ratio of GSSG/GSH were higher and the JNK was activatedin APAP-induced group compared with that in control group. However, compared to APAP-induced group, GS combined with APAP-induced group displayed a decrease of protein expression levels of ASK1, P-ASK1 and P-JNK, a reduction of serum levels of ALT and AST, a decrease in TNF-α, IL-6 and IL-1β mRNA levels, and a low ration of GSSG/GSH. Conclusions: GS-459679 treatment effectively down-regulates ASK1 and P-ASK1 expression. Addition of GS-459679 decreases the generation of liver metabolites and inflammatory factors, reduces oxidative stress reaction, inhibits JNK activation, and then protects the responsiveness to APAP-induced liver injury.

Objective To develop and verify a medical microdevice for analyzing the three-dimensional(3D)migration of tumor cells in extracellular matrix. Methods The mold of the microdevice was made by precision machining,and then the medical microdevice based on polydimethylsiloxane(PDMS)-glass was obtained by PDMS casting,moulding,and bonding.During the analysis,the suspension of tumor cells and matrigel were mixed and then added into the migration channel of microdevice,and the controllable migration of tumor cells in matrigel was induced by establishing chemokine concentration gradient on both sides of the migration channel.Meanwhile,the migration process of tumor cells was recorded with the live cell dynamic imaging device. Results Breast cancer cell line MCF-7 was taken as an example to verify the feasibility of the microdevice to control and dynamically monitor the 3D migration process of tumor cells in vitro.Qualitative analysis of imaging data showed that the migration of MCF-7 cell lines in matrigel was determined by the concentration gradient distribution direction of chemokine and presented as the amoeboid-like migration mode.The proportion and migration velocity of MCF-7 cells could be quantified by the quantitative analysis of cell migration process.The inhibition ability of matrix metalloproteinase inhibitors(Batimastat)and adenosine triphosphatase inhibitors(Blebbistation)on the 3D migration behavior of MCF-7 cells was found to be different.Conclusion This device can be used for in-depth analysis of tumor cell migration and its mechanism and for evaluating the efficacy of anti-metastatic drugs.
Biomedical Research ; instrumentation ; Cell Movement ; Extracellular Matrix ; Humans ; MCF-7 Cells

OBJECTIVE:To establish a RP-HPLC analytical method for the determination of pazufloxacin mesilate in human plasma and urine.METHODS:The plasma proteins were precipitated with methanol and the supernatant liquid ob-tained from the serum centrifugate was subjected to chromatographic analysis.The supernatant liquid obtained from the diluted urine centrifugate was subjected to sample introduction.The analytical column was Diamonsil C 18 ,the mobile phase consisted of acetonitrile-0.05mol/L potassium dihydrogen phosphate(containing1%tetrabutylammonium bromide)(8∶92,V/V)with a flow rate at1.4ml/min,excitation wavelength at320nm and emission wavelength at400nm.RESULTS:The linear range of pazufloxacin mesilate in both plasma and urine was31.25～10000ng/ml(r=0.9999).The relative recoveries of pazu-floxacin mesilate in human plasma and urine were97.77%～99.87%and98.31%～100.82%,respectively with RSD less than1.0%～3.0%.CONCLUSION:This method is accurate,reliable and simple and it is suitable for the pharmacokinetic study and routine monitoring of blood concentration of pazufloxacin mesilate.

OBJECTIVE: To develop a new method for evaluating the inhibitory effect of aspirin on platelet by the three-dimensional (3D) morphological parameters. METHODS: The sodium citrate-anticoagulant peripheral blood samples collected from 12 healthy volunteers were divided into 2 groups: group treated with 200 μmol/L acetylsalicylic acid (ASA), and control group. The platelets in the 2 groups were washed and purified. The purified platelets were added into reaction pools modified with type I collagen and induced to activation and aggregation under static condition. The 3D morphology of the formed platelet aggregate was measured by the laser shape microscopic system. Meanwhile, the platelet function was detected by turbidometric light transmittance aggregometry (LTA). RESULTS: This technology could acquire the shape, height and 3D morphology of the platelet aggregates without label, and could quantify their volume parameters. ASA treatment could obviously change the morphology of platelet aggregates. Compared with the parameters of control group, the volume of platelet aggregates in experimental group decreased significantly (t=8.97, P<0.01), while the cross-sectional area showed no significant change (t=1.94, P>0.05). The receiver-operating characteristic curve(ROC) analysis showed that the platelet aggregate volume as a parameter to identify the ASA inhibition effect had 91.7% sensitivity and 75% specificity when the cut-off value equal to 1395 μm, and its accuracy and sensitivity were both better than those of platelet aggregates rate measured by LTA method. CONCLUSION The new method developed for evaluating the ASA inhibition of platelet aggregation may provide a complementary strategy for researching and clinically evaluating of ASA anti-platelet aggregation in future.
Aspirin ; Blood Platelets ; Hemostasis ; Humans ; Platelet Aggregation ; Platelet Function Tests