Objective To prepare a redox-responsive doxorubicin-loaded nanoparticle,and to study its in vitro realease behavior and targeting effect on heptoma cells.Methods Cystamine was grafted on the side chains of hyaluronic acid with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide catalyst,and then β-cyclodextrin (β-CD) was conjugated on the amine groups of the cystamine by Schiff's base reaction to prepare β-CD modified hyaluronic acid (HACD).The HACD/DOX nanoparticles were prepared by encapsulating DOX into HACD using dialysis method.The drug loading,encapsulation efficiency,particle size and distribution,zeta potential and other physical and chemical properties,as well as in vitro drug release behavior of the HACD/DOX nanoparticles were characterized.The cytotoxicity of HACD/DOX nanoparticles to HepG2 cells was studied by cell counting kit-8 (CCK-8) method.The targeting effect of HACD/DOX nanoparticles on HepG2 cells was studied using flow cytometry and confocal laser scanning microscopy (CLSM).Results HACD were successfully synthesized,which could carry DOX to form uniform homogeneous nanoparticles.The drug loading of DOX in the nanoparticles was (16.1±0.2)％ and the encapsulation efficiency was (64.2±0.9)％.The transmission electron microscope images indicated that the shape of the HACD/DOX nanoparticles was homogeneous sphere.The results of granularity analysis showed that the average size of the HACD/DOX nanoparticles was (203.1 ±2.5) nm with a narrow size distribution (PDI =0.202).The zeta potential of the HACD/DOX nanoparticles was (-29.1±0.8) mV.The in vitro release behavior of the nanoparticles exhibited obvious redox-sensitivity.The results of in vitro cytotoxicity showed that the blank carrier material HACD had no obvious toxicity to hepatoma cells,and the HACD/DOX nanoparticles could effectively kill hepatoma cells with the 0.38 μg/ml half maximal inhibitory concentration (IC50) value at 48 h.Flow cytometry and CLSM results demonstrated that the HACD/DOX nanoparticles could target hepatoma cells through the mediating effect of hyaluronic acid.Conclusions The prepared HACD/DOX nanoparticles have suitable particle size,high drug loading and encapsulation efficiency,and can release DOX under the stimulation of reducing agent.These nanoparticles have obvious targeting effect on hepatoma cells,which is expected to be applied as the drug delivery system of hepatocellular carcinoma (HCC) therapy.

Background Chemical crosslinking agent can be used to strengthen the intensity of sclera tissue,but the intensity of the sclera may be influenced by different crosslinking methods.Objective The aim of this study was to compare the effectiveness of collagen crosslinking on porcine sclera between whole-eye crosslinking method and scleral strip crosslinking method.Methods Whole-eye crosslinking or sclera strip crosslinking was performed on 70 fresh porcine eyeballs in five groups using 1％ genipin,1％ glutaraldehyde or PBS respectively for 40 minutes.After crosslinking,10 sclera strips with l0 mm×4 mm from the temporal lateral were prepared in every group for the stress-strain measurement using a Instron5848 microtester,and the other 4 scleral strips in each group were extracted for the thermal shrinkage temperature test.Results Biomechanical property test reveled that the elastic modulus value of sclera strips reduced by 70.0％-82.8％ in the whole-eye crosslinking method group compared with scleral stip crosslinking method group after treated with 1％ genipin ((8.98 ± 1.81) MPa vs.(10.85 ± 1.83) MPa,t =3.375,P =0.003)) and 1％ glutaraldehyde((12.78 ±2.91) MPa vs.(18.25 ±5.16) MPa,t =4.007,P =0.001)) ;The tensile stress of whole-eye crosslinking method group was 54.9％-90.1％ of scleral stips method group,showing significant decline after corsslinked of whole-eye in 5％,10％,15％ and 20％ strain conditions (all P ＜ 0.05).Thermomechanics test showed that the thermal shrinkage temperature was lower in the whole-eye crosslinking group compared with scleral stip crosslinking group after treated with both 1％ genipin ((68.8 ±0.9)℃ vs.(74.8± 1.3)℃,t=11.129,P=0.000)) and 1％ glutaraldehyde((73.3±0.9)℃ vs.(79.3±1.3)℃,t=11.112,P=0.000)).Conclusions Different crosslinking methods have an influence on the efficacy of collagen crosslinking on porcine sclera.Sclera strip crosslinking offers a better crosslinking intensity for selera.

Objective To investigate the susceptibility of amphotericin B,itroconazol and voriconazole against filamentous fungi.Methods Etest was used to determine the MIC of amphotericin B, itroconazol and voriconazole against filamentous fungi including Aspergillus,Penicillium,Alternaria alternate,Mucor and Rhizopus species.Results The average MIC of voriconazol,amphotericin B and itroconazol against Aspergillus fumiagtus is O.29 ?g/ml,1.16 ?g/ml and 5.88 ?g/ml;the average MIC of amphotericin B and voriconazol to Aspergillus flavus is 6.39 ?g/ml and 0.22 ?g/ml;the average MIC of voriconazol,amphotericin B and itroconazol against Aspergillus niger is 0.69,2.31,and 19.75 ?g/ ml.Most of Penicillium are susceptable to amphotericin B,but 3 strains showed very high MIC to voriconazol and itroconazol.Both of the testing strains of Mucor and Rhizopus were resistant to all of the three antifungal agents.Conclusion Amphotericin B,itroconazol and voriconazole possessed different susceptibility on different types of filamentous fungi.It is important for clinical laboratories to identify the filamentous fungi to the level of genus and species to help physicians choose antifungal agents correctly.

Adult ; Diagnosis, Differential ; Female ; Humans ; Keratin-18 ; metabolism ; Mucin-1 ; metabolism ; Omentum ; metabolism ; pathology ; Peritoneal Neoplasms ; metabolism ; pathology ; ultrastructure ; Pregnancy ; Trophoblastic Neoplasms ; metabolism ; pathology ; ultrastructure ; Trophoblastic Tumor, Placental Site ; pathology ; Vimentin ; metabolism

Background Vitreous hemorrhage in long-term produces toxic substances and influent the metabolism of eye tissue.Lens capsule is found more thin and transparent in the eyes with chronic vitreous hemorrhage.To research the effect of vitreous hemorrhage to lens is very important for the choose of the phaco operative timing.Objective The aim of this work was to investigate the ultrastructural change of lens epithelial cells(LECs) in the eyes with experimental vitreous hemorrhage.Methods The autologous blood of 0.1 ml was intravitreally injected in the left eyes of 8 general New Zealand white rabbits,and the equal amount of phosphate buffered saline(pH7.4) was used at the same way in the right eyes.Vitreous and fundus were examined with direct ophthalmoscope on 1,3,5,9,15,20,25,30 days to assess the inflammatory response after intravitreal injection.The specimens of lens anterior capsule were obtained in 30 days after injection and the ultrastructure and apoptosis of LECs were evaluated under the transmission electron microscope. Results No obvious ocular inflammatory response was seen throughout the experimental duration,and there was no vitreous hemorrhage in the right eyes after intravitreal injection.The vitreous hemorrhage agglutinated with clear boundary in the left eyes on 1 day after intravitreal injection,and the hemorrhage turned into dark-red color on the fifth day.On the fifteenth day after injection,the hemorrhage mass became to be grey color and the vitreous liquefaction occurred in the left eyes.The hemorrhage disappeared until 25 days.But in the one month after injection of self-blood,the vitreous showed the deeper red color.The early apoptosis appeared in the LECs of the left eyes in the thirty day,presenting the enlargement and broaden of intercellular space,the decrease of mitochondria number,vacuolar change,expanse of endoplasmic reticulum and disappearance of the nuclear membrane structure. Conclusions Vitreous hemorrhage leads to the ultrastructural pathological changes of lens.

Background The pathogenesis and development of cataract is associated with oxidative stress-induced apoptosis of human lens epithelial cells(LECs).BH3-only protein is a factor that can initiate apoptosis,and thus the apoptotic process is probably related to the activation of the c-Jun N-terminal kinase(JNK).However,the relationship between oxidative stress-induced apoptosis of human LECs and the JNK pathway remains to be illuminated.Objective This study was to investigate the effects of the JNK/c-Jun pathway and its target gene,Bim (Bcl-2 interacting mediator of cell death)and PU M A(p53 up-regulated modulator of apoptosis),on oxidative stressinduced apoptosis of human LECs.Methods The human LECs cell line(HLEC-B3)was cultured and passaged in DMEM with 10％ fetal bovine serum in vitro.Confluent cells were incubated in 24 well plates and divided into 4 groups.Hydrogen peroxide(H2O2)(50 μmol/L)was used to treat the cells for 4,8 or 12 hours,and cells without H2O2 treatment served as the control group.Apoptosis was detected using Hoechst 33258 staining and quantified by counting the number of cells with pyknotic nuclei.In addition,confluent cells were seeded in 6 well plates,and Western blot and RT-PCR were used to detect the expression of the caspase-3,c-Jun,Bim and PUMA proteins and their mRNA in HLEC-B3,respectively.The JNK/c-Jun pathway inhibitors,CEP11004 or SP600125,were added into cultured media with H2O2,and cells treated with DMSO or H2O2 only served as negative and positive control groups.The expression of the p-JNK,JNK,p-c-Jun,c-Jun,Bim,PUMA proteins was detected by Western blot and apoptosis was assayed using Hoechst 33258 staining.200 pmoL/L of Bim or PUMA small interference RNA(siBim or siPUMA)fragments were transfected into the cells for 24 hours,respectively,and H2O2 was then used to treat the cells for 8 hours.The expression of the Bim and PUMA protein and their mRNA in the cells was detected by Western blot and RT-PCR,respectively.Results After H2O2 treatment in HLEC-B3 cells for 4,8,or 12 hours,the rates of apoptosis were 4.30％±1.15％,27.08％±0.74％ and 46.59％±0.91％,showing a significant difference among them (F=1909.433,P=0.000),and those of the 4,8,12 hour groups were significantly increased in comparison to the control group(P =0.049,0.000,0.000).Compared to untreated cells,the levels of expression of the JNK,Bim,PUMA proteins and their mRNA in HLEC-B3 cells were significantly elevated.After the addition of CEP11004 or SP600125,the expression of these protein and mRNA in HLEC-B3 cells in the presence of H2O2 was significantly weaker than that in the DMSO control group(P =0.000,0.000).After the tranfection of siBim or siPUMA,the apoptosis rates of the H2O2 treated groups were significantly higher than those in the Bim-/-or PIMA-/-group (P＜0.05).Conclusions H2O2 can activate the JNK/c-Jun pathway and up-regulate the expression of its target genes Bim and PUMA in human LECs in a time-dependent manner.Inhibiting the JNK/c-Jun pathway and interfering with the expression of Bim and PUMA can protect human LECs against oxidative stress-induced apoptosis.

The adverse reaction monitoring is important in warning the risks of traditional Chinese medicines at an early stage, finding potential quality problems and ensuring the safe clinical medication. In the study, efforts were made to investigate the risk signal mining techniques in line with the characteristics of traditional Chinese medicines, particularly the complexity in component, processing, compatibility, preparation and clinical medication, find early risk signals of traditional Chinese medicines and establish a traditional Chinese medicine safety evaluation system based on adverse reaction risk signals, in order to improve the target studies on traditional Chinese medicine safety, effective and timely control risks and solve the existing frequent safety issue in traditional Chinese medicines.
China ; epidemiology ; Drug Evaluation ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; etiology ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Humans ; Product Surveillance, Postmarketing

Post-marketing evaluation is a process which evaluate the risks and benefits of drug clinical application comprehensively and systematically, scientific and systematic results of post-marketing evaluation not only can provide data support for clinical application of traditional Chinese medicine, but also can be a reliable basis for the supervision department to develop risk control measures. With the increasing demands for treatment and prevention of disease, traditional Chinese medicine has been widely used, and security issues are also exposed. How to find risk signal of traditional Chinese medicine in the early stages, carry out targeted evaluation work and control risk timely have become challenges in the development of traditional Chinese medicine industry.
Drug Evaluation ; methods ; Drugs, Chinese Herbal ; Humans ; Medicine, Chinese Traditional

AIMTo investigate the inclusion of coenzyme Q10 with beta-cyclodextrin (beta-CD).

METHODSThe inclusion of the electroactive guest molecule coenzyme Q10 with the host molecule beta-CD was studied by the polarography. The change of the reduction peak current of the inclusion complex with time and the change of the peak potential of the inclusion complex with beta-CD concentration were examined. In order to study the photostability, the change of the reduction peak current of both coenzyme Q10 and coenzyme Q10-beta-CD inclusion complex with time were also examined under light, separately.

RESULTSIn 0.1 mol x L(-1) HAc/NaAc (pH 4.7) buffer-ethanol/water (60:40) medium, coenzyme Q10 was included with p-CD to form an 1:1 inclusion complex. The formation constant Kf was 1.26 x 10(4) L x mol(-1) the apparent formation rate constant was 6.64 x 10(-2) min(-1). The photodegradation apparent rate constant of coenzyme Q10 as 7.77 x 10(-3) min(-1) and that of the coenzyme Q10-beta-CD inclusion complex was 3.38 x 10(-3) min(-1).

CONCLUSIONThe inclusion of coenzyme Q10 with beta-CD took place. The stability of coenzyme Q10 to lights was improved in a certain degree due to the formation of the inclusion complex.

Coenzymes ; chemistry ; Drug Compounding ; methods ; Light ; Oxidation-Reduction ; radiation effects ; Polarography ; methods ; Ubiquinone ; analogs & derivatives ; chemistry ; beta-Cyclodextrins ; chemistry

Histocytochemistry ; methods ; Humans ; Pathology, Clinical ; methods ; Specimen Handling ; methods ; Staining and Labeling ; methods