Natural biocompatible nanomaterials such as self-assembled triterpene natural small molecule products with favorable anticancer activity show great potential for biomedical application. However, the mechanisms of their molecular self-assembled structures have not been investigated systematically, while there are still few reports of the natural active carrier for drug delivery. Herein, in this work, we further explored the molecular assembly mechanism and common regularity of tetracyclic triterpenes ergosterol, stigmasterol as well as pentacyclic triterpenes glycyrrhetinic acid and ursolic acid, which suggested that the coplanarity and orderliness of molecular arrangements which are speculated to be responsible for their self-assembly into the spherical, rod-like or lamellar nanostructure. Besides, ergosterol (ET) with better anticancer activity was chosen as a representative substance for construction of the synergistic antitumor nanodrug. By intermolecular hydrogen bonding and π-π stacking, chemotherapeutic drug paclitaxel (PTX) was encapsulated into ET-PTX NPs successfully. Then, the anti-cancer efficacy of the tumor-bearing mice was evaluated according to the protocol approved by the Experimental Animal Research Center of Harbin Medical University. The resulting nanodrug exhibited excellent biosafety and enhanced in vivo anticancer activity efficiency of 52.3%, higher than free PTX (29.4%) or ET NPs (32.5%) alone, further verifying the potential medical application value of triterpene natural products. This work provides not only a theoretical basis for exploring the self-assembly behavior of small molecule natural products, but also a promising perspective for the fabrication of active natural biocompatible nanodrug delivery systems for synergistic antitumor therapy and other biomedical applications.

Animals ; Bile ; Carps ; Hemofiltration ; Humans ; Poisoning ; therapy

Objective To study the relation between the effect of Curcumin on the proliferation and activation of hepatic stellate cells(HSC)and the expression of the peroxisome proliferator-activated receptor?(PPAR?).Methods The rat HSC was isolated from SD rats through in situ perfusion of liver with Pronase E and density-gradient centrifugation with Nycodenz.The subcultured cells were treated with corresponding compound.The inhibition effect on HSC proliferation was determined by MTT colorimetry.The total RNA was extracted by TRizol reagent,and the gene expression level of PPAR? and smooth muscle actin (?-SMA)were determined by semi-quantitative RT-PCR.The total cellular proteins were extracted and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis,and the protein level of ?-SMA was determined by Western blotting.Results The MTT analysis results showed that Curcumin inhibited HSC proliferation between 10 and 50 ?mol/L in a dose-dependent manner.In the Day 1,4,7 of primary HSC and passaged HSC,the expression of PPAR? mRNA decreased with HSC activation extent;Curcumin could up-regulate the expression level of PPAR?.Curcumin suppressed the expression of ?-SMA not only at the gene level but also at the translation level.But all these effects of Curcumin on HSC could be blocked by the PPAR? specific antagonist GW9662.Conclusion The effect of Curcumin on HSC cell proliferation and activation is through the up-regulation of PPAR? signal.

Aim To study the effect of Curcumin on the apoptosis of hepatic stellate cells (HSC), and the correlation between the effect and peroxisome proliferator-activated receptor ?(PPAR?) signal.Methods The HSC was isolated from normal SD rats through in situ perfusion of liver with protease E and density-gradient centrifugation with Nycodenz.The subcultured cells were treated with corresponding compounds. Cell apoptosis was detected by Hoechst 33258 staining. PPAR? subcellular distribution was detected by immunofluorescent staining. Total RNA, total protein and nuclear protein were extracted respectively, target gene and protein levels were determined by semi-quantitative RT-PCR or Western blot.Results There was nearly no apoptosis in activated HSC. Curcumin treatment induced the apoptosis of HSC, enhancing PPAR? nuclear translocation/redistribution.At the transcription and translation level,curcumin upregulated nuclear PPAR? expression, inhibited anti-apoptotic Bcl-2 expression, and promoted pro-apoptotic Bax expression; but all these effects could be reversed by PPAR? antagonist GW9662.Conclusions Curcumin induces HSC apoptosis by enhancing PPAR? expression and nuclear translocation/redistribution.

Objective To clone Tp0821,Tp0319,Tp0971 and Tp0663 gene,construct prokaryotic expression plasmid,the ex-pression of recombinant proteins,purification and used to detect syphilis patients serum antibodies.To explore Tp0821, Tp0971,Tp0319 and Tp0663 recombinant protein in the diagnosis of Tp infection,rich library of candidate antigens syphilis serology diagnosis.Methods From Tp library screening Tp0821,Tp0319,Tp0971 and Tp0663 four kinds of membrane pro-tein,through the analysis of the bioinformatics software selection advantage antigen epitope,connected with pQE32 carrier respectively built pQE32/Tp0821,pQE32/Tp0319,pQE32/Tp0971 and pQE32/Tp0663 prokaryotic expression recombi-nant;In vitro cloning,expression and purification and on single Tp0821,Tp0319,Tp0971,Tp0663 and four kinds of recombi-nant protein chimeric antigen (Tp0821-Tp0319-Tp0971-Tp0663)in order to establish the indirect Tp-envelope antigen ELISA method,based on TRUST and TPPA method comparison,detection collected from January 2013 to June 2014 in Shenzhen Baoan District People’s Hospital outpatient and hospitalization Tp negative patients,160 cases of positive speci-mens of 83 cases of clinical and Tp,and evaluated its application in the syphilis serology diagnosis.Results Successful con-structed of prokaryotic expression vector pQE32/Tp0821,pQE32/Tp0319,pQE32/Tp0971 and pQE32/Tp0663.Efficient expressed and purified their relative molecular mass of recombinant proteins.Seted up Tp-indirect ELISA method to detect 160 cases of Tp negativepositive clinical specimens and 83 cases of Tp,and compared with TPPA,the sensitivity were 85.5% (71/83),84.3% (70/83),89.2% (74/83),81.9% (68/83)and 95.2% (79/83)respectively,specificity was 100%(160/160),and the coincidence rate was 82.6%~95.7%.Single Tp0821,Tp0319,Tp0971 and Tp0663 recombinant protein positive rate of TP-ELISA was established (85.5%,84.3%,89.2% and 81.9%)compared with TPPA method of positive detection rate (100%)had significant difference statistically significant (χ2= 24.5~31.8,P<0.01),and four kinds of re-combinant protein chimeric antigen positive rate of TP-ELISA was established with the TPPA method the positive detection rate of comparative difference was statistically significant (χ2=7.95,P<0.05).Conclusion Preparation Tp0821,Tp0319, Tp0971 and Tp0663 recombinant protein had good immune activity,for the further study of its application in the diagnosis of syphilis serology lay a certain foundation,but in four kinds of recombinant protein chimeric antigen based Tp Tp-indirect ELISA method for serological diagnosis,can improve the detection sensitivity.

Adenoviridae ; genetics ; Cell Death ; Genetic Vectors ; HT29 Cells ; Humans ; Interleukins ; biosynthesis ; genetics ; RNA, Catalytic ; biosynthesis ; genetics ; RNA, Messenger ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; metabolism

Female ; Humans ; Liability, Legal ; Middle Aged ; Occupational Diseases ; diagnosis ; Occupational Health ; legislation & jurisprudence ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; etiology

OBJECTIVE:To establish a method for the content determination of chlorogenic acid,protocatechuic acid and total phenolics in Chaenomeles sinensis,compare the content of total phenolics and 2 phenolic acids from different areas. METHODS:Wavelength switching HPLC method was conducted to determine the chlorogenic acid and protocatechuic acid. The column was Shim-pack CLC-ODS(M)with the mobile phase of acetonitrile-0.1% phoephoric acid(15∶85,V/V),the detection wavelength was 259 nm(for protocatechuic acid)and 325 nm(for chlorogenic acid)and the switching time was 14 min. With the index of proto-catechuic acid,Folin-Ciocalteau colorimetric method was conducted to determine the total phenolics. RESULTS:The 2 methods of quantitative analysis showed that the precision,repeatability,recoveries and standard curves were all validated by methodology. The mass fraction of total phenolics was 0.87%-3.77% with the average of 2.16%;the chlorogenic acid was 0.053%-0.387% with the average of 0.192% and the protocatechuic acid was 0.024%-0.541% with the average of 0.087%. The order of total phenolics content in C. sinensis from different areas was Yunnan>Anhui Xuancheng>Sichuan>Hubei and the order of total amount of chlo-rogenic acid and protocatechuic acid from different areas was the same as the total phenolics. There were differences among the con-tents of C. sinensis from different areas,however,the positive correlation was found between the content of total phenolics and the total amount of chlorogenic acid and protocatechuic acid with the pearson correlation coefficient of 0.719(P<0.01).CONCLU-SIONS:The established method is simple,accurate and reproducible and can be used for the content determination of chlorogenic acid,protocatechuic acid and total phenolics in chaenomelis fructus.

AIM:To study the effect of different surgeries for vision and postoperative complications in leprosy patients with lagophthalmos.METHODS:A retrospective analysis.Totally 68 leprosy patients (97 eyes) with lagophthalmos were selected who treated with different surgeries during May 2007 to September 2015 in our sanatorium.The surgeries included nylon thread correction surgery, tarsorrhaphy, temporalis transposition surgery.We observed and compared the best corrected visual acuity(BCVA) and the rate of postoperative complications, such as epiphora, lacrimal duct injury, hard to open eyes, infection at the inner and lateral canthus and scar formation of lacrimal apparatus.RESULTS:The effect on BCVA of patients treated with nylon thread correction surgery and tarsorrhaphy:the preoperative and postoperative BCVA was significantly different of the two groups (Z=-4.193,-4.213;P<0.05);the preoperative and postoperative BCVA was not significantly different of the patients underwent temporalis transposition surgery (Z=-1.532, P >0.05).The postoperative complications:(1) epiphora:there was 91% in patients underwent nylon thread correction surgery, 89% in patients underwent tarsorrhaphy, 59% in patients underwent temporalis transposition surgery, the difference was statistically significant (χ2=12.198, P<0.05);(2) lacrimal duct injury:there was 66% in patients underwent nylon thread correction surgery, 95% in patients underwent tarsorrhaphy, 41% in patients underwent temporalis transposition surgery, the difference was statistically significant (χ2=22.415, P<0.05);(3) hard to open eyes:there was 56% in patients underwent nylon thread correction surgery, 55% in patients underwent tarsorrhaphy, 22% in patients underwent temporalis transposition surgery, the difference was statistically significant (χ2=8.795, P<0.05);(4) infections:there was 84% in patients underwent nylon thread correction surgery, 3% in patients underwent tarsorrhaphy, 11% in patients underwent temporalis transposition surgery, the difference was statistically significant (χ2=60.858, P<0.05).CONCLUSION:There are certain clinical efficacy for lagophthalmos caused by leprosy after surgery, especially patients after temporalis transposition surgery can close eyes by oneself.And their vision does not change significantly, which suggests that this surgery may prevent exposure injury for cornea.But the iatrogenic injury, the vision decrease in patients underwent other surgeries and the ocular tissue damage, such as epiphora, lacrimal duct injury, hard to open eyes, infection at the inner and lateral canthus and scar formation of lacrimal apparatus should be pay more attention to.