Objective To study the differential proteome of kidney between lupus nephritis mouse and normal mouse.Methods The proteins of kidney were separated by two-dimensional gel electrophoresis(2-DE).The gels stained by silver were scanned by ImageScanner and analyzed by PDQuest software.Results About 573?52 and 658?43 protein spots were found in the three maps of control group and LN group respectively;the match ratio was 83% and 87% respectively.One hundred and fourteen spots were found increased that showed a two fold increase as comparing to control group.Conclusion A significant difference in protein expression of LN mouse kidney was found and may be related to the pathogenesis of LN.

Objective To investigate the effect of norcantharidin on growth inhibition and induction of apoptosis of human rectal cancer Colo 320 cells. Methods Norcantharidin (NCTD) in different concentrations were added to rectal cancer Colo 320 cells. Morphological characteristics of apoptosis were observed using the light microscope and transmission electron microscope. The expressions of Bag-1 and Bcl-2 proteins were tested by Western blotting. The growth inhibition of Colo 320 cells on the cell cycle was observed by flow cytometry. Results The apoptosis morphological changes of Colo 320 cells were observed by the light microscope and transmission electron microscopy. Flow cytometry analysis showed that the cell count of G2/M phase in experimental group was higher than that in control group ( <0.05) but the cell counts of G0/G1 and S phases have decreased in experimental group after treatment with NCTD at the concentrations of 5μg/mL, 10μg/mL and 20 μg/mL, and presented dosage dependence relations. The expressions of Bag-1 and Bcl-2 proteins have decreased. Conclusion Norcantharidin has inhibitory effect on rectal cancer Colo 320 cells, and the effect may be related to the cell cycle arrest and apoptosis.

Organic anion transporting polypeptide 1B1 (OATP1B1) is an important liver-specific uptake transporter, which mediates transport of numerous endogenous substances and drugs from blood into hepatocytes. To identify and investigate potential modulators of OATP1B1 from natural products, the effect of 21 frequently used natural compounds and extracts on OATP1B1-mediated fluorescein methotrexate transport was studied by using Chinese hamster ovary cells stably expressing OATP1B1 (CHO-OATP1B1) in 96-well plates. This method could be used for the screening of large compound libraries. Our studies showed that some flavonoids (e.g., quercetin, quercitrin, rutin, chrysanthemum flavonoids and mulberrin) and triterpenoids (e.g., glycyrrhetinic acid and glycyrrhizic acid) were inhibitors of OATP1B1 with IC50 values less than 16 µmol · L(-1). The IC50 value of glycyrrhetinic acid on OATP1B1 was comparable to its blood concentration in clinics, indicating an OATPlB1-mediated drug-drug interaction could occur. Structure-activity relationship analysis showed that flavonoids had much higher inhibitory activity than their glycosides. Furthermore, the type and length of saccharides had a significant effect on their activity. In addition, we used OATP1B1 substrates fluvastatin and rosuvastatin as probe drugs to investigate the substrate-dependent effect of several natural compounds on the function of OATP1B1 in vitro. Our results demonstrated that the effect of these natural products on the function of OATPlB1 was substrate-dependent. In summary, this study would be conducive to predicting and avoiding potential OATP1B1-mediated drug-drug and drug-food interactions and thus provide the experimental basis and guidance for rational drug use.
Animals ; Biological Products ; CHO Cells ; Cricetulus ; Drug Interactions ; Fatty Acids, Monounsaturated ; pharmacology ; Flavonoids ; pharmacology ; Indoles ; pharmacology ; Inhibitory Concentration 50 ; Organic Anion Transporters ; genetics ; metabolism ; Rosuvastatin Calcium ; pharmacology ; Structure-Activity Relationship

Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma were widely used in strengthening spleen under different disease conditions, and were easily and often misused each other. Therefore, DNA barcode was used to distinguish Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from their adulterants to ensure the safe use. The sequence lengths of ITS2 of Atractylodes macrocephala, Atractylodis Rhizoma (A. lancea, A. japonica and A. coreana) were both 229 bp. Among the ITS2 sequences of A. macrocephala, only one G/C transversion was detected at site 98, and the average GC content was 69.42%. No variable site was detected in the ITS2 sequences of A. lancea. The maximum K2P intraspecific genetic distances of both A. japonica and A. coreana were 0.013. The maximum K2P intraspecific genetic distances of A. macrocephala, A. lancea, A. japonica and A. coreana were less than the minimum interspecific genetic distance of adulterants. The ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. DNA barcoding could be used as a fast and accurate identification method to distinguish Atractylodis Macrocephalae Rhizoma, Atractylodis Rhizoma, from their adulterants to ensure its safe use.
Atractylodes ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

Objective To develop the knowledge-attitude-practice scale for evaluating intra-abdominal pressure measurement in ICU nurses and assess its reliability and validity preliminary. Methods Applied the methods of literature review and Delphi expert consultation to form the knowledge-attitude-practice scale for evaluating intra-abdominal pressure measurement in ICU nurses on the basis of the knowledge-attitude-practice framework. A total of 165 nurses from ICU department were finally involved. Items analysis, exploratory factor analysis, content validity, internal consistency test and split-half reliability were used to evaluate the scale. Results Factor analysis revealed five factors (23 items), accounting for 52.5%of the total variance. The Cronbach αcoefficient was 0.869 for the total scale and 0.612, 0.749 and 0.848 for the subscales of knowledge, attitude and practice. The split-half coefficient was 0.784. Conclusions The knowledge-attitude-practice scale has good reliability and validity, and can be used to assess the intra-abdominal pressure measurement in ICU nurses.

0.05).Conclusion CYPIA1 MspⅠ polymorphism is not related to the susceptibility to colorectal carcinoma.

To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; drug therapy ; enzymology ; genetics ; physiopathology ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Female ; Humans ; MCF-7 Cells ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology ; src-Family Kinases ; genetics ; metabolism

Objective To explore the clinicopathologic characteristics,to screen risk factors of metastasis and to analyze the diagnosis,treatment and prognosis of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN).Methods From January 2010 to November 2015,the clinical data of 405 patients with GEP-NEN were retrospectively analyzed.GEP-NEN tumors were classified as neuroendocrine tumor (NET,G1 and G2 grade),neuroendocrine carcinoma (NEC,G3 grade) and mixed adenoendocrine carcinoma (MANEC,G3 grade).The clinicopathologic characteristics were summarized.The staining characteristics of synaptophysin (Syn),chromogranin A (CgA) and CD56 of tumor tissues were analyzed by immunohistochemistry.x2 and t test were performed to analyze differences in pathologic characteristics between groups.Logistics regression method was used to analyze the risk factors of metastasis.KaplanMeier method and Log-rank test were used for survival analysis.Results The mean age of patients with GEP-NEN was (54.7± 13.3) years.Gastric NEN was the most common GEP-NEN (98 cases,24.2％),followed by 95 cases (23.5％) with NEN in rectum,86 cases (21.2％) in pancreas and 50 cases (12.3％)in esophagus.Among them,47 cases (11.6％) were functional GEP NEN and 358 cases (88.4％) were non-functional GEP-NEN.According to pathologic diagnosis,227 cases (56.0％) were NET,125 cases (30.9％) were NEC and 16 cases (4.0％) were MANEC.According to tumor classification,120 cases (29.6％) were grade G1,108 cases (26.7％) were grade G2 and 177 cases (43.7％) were grade G3.Immunohistochemistry staining positive ratesof Syn,CgA and CD56 were 97.4 ％ (381/391),44.0 ％ (121/275) and 83.9％(291/347),respectively.The median (lower quartile,upper quartile) diameter of grade G1,G2 and G3 tumors were 1.0 cm (0.6 cm,1.5 cm),1.5 cm (1.0 cm,2.5 cm),4.0 cm(2.5 cm,6.0 cm),respectively,and the difference was statistically significant (Z =99.171,P ＜ 0.01).The positive rate of CgA of grade G3 tumor was lower than that of grade G1 and G2(x2 =7.078 and 11.391,both P＜ 0.01).The results of Logistic regression analysis revealed that tumor size and pathologic classification were the important predictors of metastasis.The median survival time of metastasis group and non-metastasis group of grade G3 was 12.0 months and 41.5 months,and there was a significant difference between the two groups by Log-rank test (x2 =37.075,P＜0.01).Conclusions GEP-NEN may occur at any part of the digestive system.There are differences in tumor size positive rate of,immunohistochemistry staining and the primary site of tumors with different pathological grading.The tumor diameter and pathologic classification are the important predictors of metastasis.The prognosis of metastasis group is worse than that of non-metastasis group.

Exosomes are a kind of endosomal vesicles that are secreted by most if not all living cells. Due to their capability of delivering a variety of cargos, such as tissue- or cell-specific proteins, lipids, and genetic materials, and their broad biological activities, exosomes have gained substantial attention as emerging therapeutics. Exosomes derived from mesenchymal stem cells (MSCs) and dendritic cells (DCs) are two types of exosomes that are widely studied. Many preclinical and clinical studies have shown that they have a satisfactory treatment effect in lung diseases, liver diseases, nervous system diseases, tumors, and other diseases. In addition, exosomes from macrophages, tumor cells, plant cells, and many other cells are getting more attention due to their therapeutic potential. Besides natural exosomes, research on engineered exosomes has also made plenty of progress. There have been several engineering methods of exosomes, such as targeting modification and loading of active ingredients. In this review, we summarize the research progress of therapeutic exosomes from different sources, and further discusses the application prospects of exosomes and possible challenges in the future.

In order to establish an efficient and low-cost production procedure of recombinant glycerol kinase (r-GK), we expressed the r-GK gene at high level in E. coli by induction with lactose on a large-scale fermentation of 300L. The results showed that the biomass concentration reached OD600 of 42 and the expression of r-GK in E. coli accounted for about 30% of total soluble protein. The cell-free extract was processed by selective thermo-denaturation and then purified with Ni sepharose FF column chromatography. Finally, highly purified r-GK was obtained and its purity reached 97% by using analysis on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), polyacrylamide gel electrophoresis (PAGE) and gradient polyacrylamide gel electrophoresis (Gradient PAGE). Further identification study showed that the molecular weight of r-GK was 120kDa with two subunit of 58kDa. Contaminants of NADH oxidase and catalase were not detected in the sample pool of r-GK. The purified r-GK was able to retain about 85% of its initial activity at 4 degrees C for 30 days. After lyophilized, it can retain 93% of its initial activity at 4 degrees C for one year.
Escherichia coli ; genetics ; metabolism ; Fermentation ; Glycerol Kinase ; biosynthesis ; genetics ; isolation & purification ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification