Objective To evaluate the protective effect of curcumin against reperfusion injury of mouse renaltubular epithelial cells in vitro and the related mechanisms. Methods The renal tubular epithelial cells(TECs)were isolated by an improved Percoll density gradient centrifugation method. The cultured TECs were divided into blank group, hypoxia reperfusion group, low-concentration curcumin (5μmol/L) and high? Concentration curcumin (25μmol/L) groups. Apoptoticrate of TECs was examined with FACS, and them RNA and protein levels of TLR4 were detected with RT-PCR and Westernblotting analysis, respectively. Meanwhile, expression of IL-6, IL-1ß, TNF-α and MCP-1 mRNA in TECs was examined by RT-PCR, and their contents in the supernatants of TECs were measured by ELISA. Results The purity of the isolated TECs washigher than 95%, and the cell viability was higher than 93%. Compared with hypoxia reperfusion group, the two curcumin groups exhibited significantly lower apoptotic rates (P<0.01) and lower levels of TLR 4m RNA and protein (P<0.01), inhibited P65 phosphorylation, and significantly decreased levels of IL-6, IL-1ß, TNF-α and MCP-1 in the TECs (at both mRNA and protein levels) and their supernatant (P<0.05orP<0.01). Conclusion Curcumin can protect TECs from reperfusion injury in vitro, which might be associated with the inhibition of TLR4 expression and the consequent attenuation of TLR4- triggered inflammatory damages.

Objective To learn the gene mutation type and prevalence of thalassemia in the Taishan area and to provide basis for the prevention and treatment of local thalassemia .Methods 1 383 patients who visited the hospital for thalassemia genetic test from January to December 2014 were enrolled for this study .α‐andβ‐genes were determined by using PCR and membrane hybridization methods .Results Of the 1 383 patients ,595 were diagnosed with thalassemia (positive rate was 43 .02% ) ,including 386 cases (27 .91% ) of α‐thalassemia ,183 cases(13 .23% ) of β‐thalassemia and 26 cases(1 .88% ) of αβ‐thalassemia .- -SEA/αα(accounted for 64 .25% ) was the major genotype of α‐thalassemia ,the major genotype of β‐thalassemia wasβ41 -42/β(accounted for 28 .42% ) , and - -SEA/αα,β41 -42/βwas the major genotype of αβ‐thalassemia (accounted for 19 .23% ) .Conclusion Taishan is located in a high risk area of thalassemia ,it is of great significance to perform genetic diagnosis of thalassemia among reproductive population so as to reduce the birth rate of children with thalassemia in the area .

OBJECTIVEThis study aims to determine the effect of fluoride concentration on the corrosion behavior of cobalt-chromium alloy fabricated by two different technology processes in a simulated oral environment.

METHODSA total of 15 specimens were employed with selective laser melting (SLM) and another 15 for traditional casting (Cast) in cobalt-chromium alloy powders and blocks with the same material composition. The corrosion behavior of the specimens was studied by potentiodynamic polarization test under different oral environments with varying solubilities of fluorine (0, 0.05%, and 0.20% for each) in acid artificial saliva (pH = 5.0). The specimens were soaked in fluorine for 24 h, and the surface microstructure was observed under a field emission scanning electron microscope after immersing the specimens in the test solution at constant temperature.

RESULTSThe corrosion potential (Ecorr) value of the cobalt-chromium alloy cast decreased with increasing fluoride concentration in acidic artificial saliva. The Ecorr, Icorr, and Rp values of the cobalt-chromium alloy fabricated by two different technology processes changed significantly when the fluoride concentration was 0.20% (P < 0.05). The Ecorr, Icorr, and Rp values of the cobalt-chromium alloy fabricated by two different technology processes exhibited a statistically significant difference. The Icorr value of the cobalt-chromium alloy cast was higher than that in the SLM group cobalt-chromium alloy when the fluoride concentration was 0.20% (P < 0.05). The Ecorr, tRp alues of the cobalt-chromium alloy cast were lower htan those of the SLM group cobalt-chromium alloy when the fluoride concentration was 0.20% (P< 0 .05).

CONCLUSIONFluoride ions adversely affected the corrosion resistance of the cobalt-chromium alloy fabricated by two different technology processes. The corrosion resistance of the cobalt-chromium alloy cast was worse than that of the SLM group cobalt-chromium alloy when the fluoride concentration was 0.20%.

Chromium Alloys ; Corrosion ; Fluorides ; Lasers ; Phosphates ; Saliva, Artificial ; Sodium Fluoride

Objective To investigate the correlationship between depressed behaviors and interleukin-1β (IL-1β) in brain tissue in mice which are insensitive to fluoxetine,and to mimic the treatment resistant depression (TRD) in clinical condition.Methods 50 BALB/c mice were randomly divided into Control group (Control),Chronic unpredictable mild stress (CUMS) group and CUMS+fluoxetine group.Mice in Control group were raised ad libitum for 9 weeks,those in CUMS group received CUMS for 9 weeks and those in CUMS+fluoxetine group received 8 weeks' CUMS followed 1 week' s treatment with Fluoxetine(10 mg · kg-1 · d-1).At the end of the 9th week,mice in(CUMS + treatment)group were selected into antidepressant treatment-resistant mice(ATRM) as no remission and Depression Group (DM) as symptoms improved.Body mass test (BMT),open field test (OFT) and forces swim test (FST) were completed respectively in these 4 groups at the endpoint of the experiment,and the brain tissue were extracted after the tests for IL-1β Elisa test.Results (1) BMT:there was no effect of weightgain in ATRM after 1 week' s therapy with Fluoxetine.There was no difference in body-weight between ATRM ((18.56±7.56) g) and CUMS ((19.03± 8.58) g) mice,while compared with Control ((24.56±5.45) g) and DM mice ((20.12±9.17) g) ATRM and CUMS mile's body weight were significantly lower (P＜0.05).(2)OFT and FST:in OFT,there was no significant difference in of horizontal moving distance(F=0.355) either in the frequencies of entering the central zone (F=0.327) among the 4 groups;in OFT,the immobility time of ATRM ((241.50 ± ± 36.55) s) was significantly longer than that in DM ((156.00± 25.47) s) (F=13.573,P＜0.05).(3) Elisa test of IL-1β:the brain' s IL-1β serum level in ATRM ((164.90±46.70) pg/mg) was higher than those in Control ((69.68±6.56) pg/mg)),and DM ((93.09±4.65) pg/mg) (P＜0.01),while no difference with that in CUMS mice.Additionally,the depressive behaviors in ATRM showed its positive correlation with the IL-1β level in CNS (r=0.669,P=0.006).Conclusion CUMS can elicit the refractory depressive symptoms in BALB/c mice to simulate TRD' s characteristic,and the elevated level of IL-1 β within brain tissue may play an important role in the development of TRD.

Objective:To explore whether apoptotic ovarian cancer cells induced by chemotherapy drugs paclitaxel and cisplatin can be cross-presented by dendritic cells(DCs) and enhance immune responses.Methods:DCs were induced from peripheral blood monocytes cells by GM-CSF/IL-4 for 6 d,then they were stimulated with either apoptotic ovarian cancer HO8910 cells,frozen-thawed HO8910 cells or control cells for 4 h.Their surface markers and phagocytotic ability were detected by flow cytometry and confocal microscopic scanning assay,respectively.DCs of different groups were cultured with CD8+ T cells isolated by magnetic cell sorting,and the ability of DCs to activate CD8+ T cells was evaluated by 3H-TdR,the activity of CTL to kill tumor cells was evaluated by LDH.Production of IFN-? by CD8+ T cells was measured by ELISPOT.Results:Apoptotic ovarian cancer cells induced by chemotherapy drugs paclitaxel and cisplatin could be phagocytized by DCs,which subsequently promoted the maturation and antigen presenting ability of DCs.Apoptotic ovarian cancer cells implused DCs significantly promoted proliferation of CD8+ T cells compared with that of control cells(P

Background As a main suppressing neurotransmitter in visual system,gamma-aminobutyric acid (GABA) participates in the transmission and regulation of visual information.GABA and dopamine (DA) coexist in the visual cortex,and their mutual effects should be clarified.Objective This study was to investigate the effect of DA on GABA-activated current in vitro in cultured visual cortical neurons of rats.Methods Neutrons from visual cortex of clean neonatal rats were isolated and cultured by explant culture method.The neutrons cultured for 11 -to 14- days were collected for the record of whole cell currents of GABA-A (IGABA) channels in vitro using patch clamp technique.The DA solution (100mmol/L),SKF38393 (10mmol/L) solution and quinpirole solution(10mmol/L) were prepared with double distilled water and then the extracellular fluid was added to different concentrations.The IGABA changing rate under the action of SKF38393+SCH23390,SKF38393+Quinpirole for different time was recorded respectively and compared with that of action of DA,SKF38393,SCH23390,Quinpirole.The IGABA activated by extracellular fluid along served as control.Results The IGABA was significantly attenuated after activated by ≥10μmol/L of DA or SKF38393 separately in comparison to that of extracellular fluid action (P＜0.05).In various time of action,there were obviously differences in IGABA between DA or SKF38393 action and extracellular fluid (P＜0.05,P＜0.01).No evident change in IGABA changing rate was found after only SCH23390 action in comparison with extracellular cells (P＜0.05).However,after combination of SCH23390 and SKF38393,IGABA changing rate reduced by 19.49%.No significant differences were found in the changing rates of IGABA among different concentrations of quinpirole action groups compared with extracellular fluid group (P＞0.05);while when quinpirole was combined with SKF38393,the IGABA was elevated in comparison with only SKF38393 action group (P＜0.05).Conclusion Dopamine participates in the transfer and regulation of visual information through suppressing GABA-activated currents from neutrons of visual cortex at time-dependent manner in vitro.

Objective To explore immunotherapy effective combined with active immunotherapy in different time according rats bearing-tumor after paclitaxel and carboplatin chemotherapy, and to identify the optimization time and strategy of vaccine and seek rational chemo-immunotherapy strategies in ovarian cancer treatment. Methods The dynamic immunocytes number and function in established tumor treated with paclitaxel and carboplatin chemotherapy were investigated. The changes of established tumor volume and immune function of different groups were observed according to combining different time after chemotherapy and vaccine. Results Lymphopenia was observed and the number of lymphocyte subset decreased remarkably on the 6th day, but all cells were found almost recovered on 15th day after chemotherapy. There is the process of immune-enhancing from post-chemotherapy 6 day to 10 day and reversal of immune suppression temporary. The combination post-chemotherapy 6 day with CTL caused a significantly delayed tumor growth in both tumor models and induced significant the proliferation of T lymphocyte by [H] 3 releasing. The number of CD8+T cell is the highest, but the expression of Tr cell was lowest in the group of post-chemotherapy 6 day with CTL. Furthermore, the ability of CD8+T secretion IFN-γ is the most in the post-chemotherapy 6 day with immunotherapy groups. Conclusion Combinational paclitaxel and carboplatin chemotherapy has synergistic effects with active immunotherapy boosting against tumor during window periods, where 6 days after chemotherapy with the most decreased number of lymphocytes in the animal periphery might represent the optimal checkpoint for the immune therapy against tumors. Therefore, monitoring the immune status of tumor patients might become one of the important prerequisites for the effective immune therapy when designing the comprehensive therapeutic strategies.

Objective To investigate the effect of rosmarinic acid on the behavioral changes in enhanced single prolonged stress (ESPS) model rats and the levels of interlukin-1β (IL-1β) and interlukin-6 (IL-6) in the hippocampus.Methods 48 adult male Sprague Dawley rats were randomly divided into six groups (n =8):Control group,Control + RA (L) group,Control + RA (H) group,ESPS group,ESPS + RA (L) group and ESPS + RA (H) group.Behavioral changes of these rats were analyzed by open field test and elevated plus-maze.The levels of IL-1 β and IL-6 were measured by enzyme linked immunosorbent assay (ELISA).Results (1) Open field test showed that the number of central region entering and the fraction of time exploring in center of ESPS group were significantly reduced than that of Control group ((18.13 ± 10.15) times,(26.68 ± 10.06) ％) and ESPS + RA (H) group ((16.88 ± 8.81) times,(25.08 ± 8.52) ％) (P ＜ 0.05).And it showed no significant difference among Control + RA(L) group,Control + RA(H) group and Control group.Meanwhile,there was also no statistic difference between ESPS group and ESPS + RA(L) group.(2) Elevated plus-maze test showed that percentages of open arm entries and fraction of time exploring in open arm in reference to total number of entries into all arms and total time spent on all arms in ESPS group were significantly reduced than that of Control group((37.38 ± 8.24)％,(17.63 ±4.74)％) and ESPS + RA(H) group((33.72 ±9.49)％,(16.99 ±4.28)％) (P ＜ 0.05),but there was no significant difference between that of ESPS group and ESPS + RA(L) group(P＞0.05).It also showed no significant difference among Control + RA (L) group,Control + RA (H) group and Control group.(3) Compared with ESPS group,RA(10mg/kg) reduced the levels of IL-1β and IL-6 in the hippocampus,but RA(5mg/kg) did not have this effect.(4) Correlation analysis results showed the level of IL-1β in the hippocampus was negatively related with the ameliorated PTSD-like behaviors of ESPS exposure rats.Conclusion RA can ameliorate PTSDlike behaviors of ESPS exposure rats,and this effect may be carried out by down-regulating the levels of IL-1β and IL-6 in the hippocampus,especially the IL-1β.

OBJECTIVETo clone and construct a eukaryotic expression vector of human bone morphogenetic protein (BMP) 2 and histidine in vitro and synthesize chitosan (CS)/pIRES2-EGFP-hBMP2-His nanoparticles.

METHODSpMD18T-hBMP2-His was digested by EcoR I and BamH I to obtain the hBMP2-His gene, which was inserted into pIRES2-EGFP to form pIRES2-EGFP-hBMP2-His. Afterward, CS, which exhibited five different molecular weights and deacetylation degrees, was complexed with pIRES2-EGFP-hBMP2-His to form CS/pIRES2-EGFP-hBMP2-His nanoparticles; in this procedure, a desolvent method was used at different N/P ratios (amino in CS to phospho in plasmid DNA). The gene-encapsulating ability of CS was evaluated by agarose gel electrophoresis and fluorescence spectrophotometry; size, distribution, and potential were analyzed using a ZetaPALS analyzer. The shape of the nanoparticles was observed under an atomic force microscope.

RESULTS1) pIRES2-EGFP-hBMP2-His was constructed after the cloned hBMP2-His gene was confirmed by sequencing. 2) CS/pIRES2-EGFP-hBMP2-His nanoparticles were synthesized and pIRES2-EGFP-hBMP2-His was packaged by CS. 3) CS/pIRES2-EGFP-hBMP2-His nanoparticles were globular with an average size of 111.7 nm to 3,214.2 nm and an average zeta-potential of 4.93 mV to 16.79 mV.

CONCLUSIONCS/pIRES2-EGFP-hBMP2-His nanospheres are successfully synthesized.

Bone Morphogenetic Protein 2 ; Chitosan ; Genetic Vectors ; Green Fluorescent Proteins ; Histidine ; Humans ; Nanoparticles ; Plasmids

AIMTo establish an HPLC method for simultaneous determination of four flavonoids in Hypericum perforatum L., ruitn, hyperin, avicularin and quercetin.

METHODSC18 column was used, the chromatography was carried out with a linear gradient program. The mobile phase was A: H2O (pH 3.0-3.5 adjusted with phosphoric acid) and B: acetonitrile, at flow rate of 1.0 mL.min-1, peaks were detected at 254 nm.

RESULTSThe linear range of rutin was 1.376-8.256 micrograms.mL-1 (gamma = 0.9999), hyperin 3.160-18.960 micrograms.mL-1 (gamma = 0.9996), avicularin 0.968-5.808 micrograms.mL-1 (gamma = 0.9998) and quercetin 0.776-4.656 micrograms.mL-1 (gamma = 0.9993). The average recovery of rutin was 97.8%, RSD 4.8% (n = 3), hyperin 100.7%, RSD 3.7% (n = 3), avicularin 97.3%, RSD 0.8% (n = 3) and quercetin 100.5%, RSD 4.4% (n = 3). All of RSDs of precision were less than 2% (n = 5), reproducibilities less than 3%.

CONCLUSIONThe method is simple, effective and feasible. It can be used to determine flavonodis in Hypericum perforatum.

Calibration ; Chromatography, High Pressure Liquid ; Flavonoids ; analysis ; chemistry ; Hypericum ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analogs & derivatives ; analysis ; Reproducibility of Results ; Rutin ; analysis