Objective: To develop the ultra performance liquid chromatography tandem mass spectrometry ( UPLC-MS/MS ) for the determination of perfluorocarboxylic acids ( PFCAs ) in fish. Methods: The fish samples were extracted with tert-butyl methyl ether and purified by WAX columns. The WAX cartridges were rinsed with methanol and 25 mmol/L ammonium acetate, and the target compound residues were eluted with 0.5% ammonia methanol and then redissolved with 50% methanol aqueous solution after nitrogen blowing to nearly dry. Nine kinds of PFCAs were simultaneously quantified by UPLC-MS/MS with 1 mmol/L ammonium acetate-methanol solution as the mobile phase. Results: The extraction was separated well in UPLC BEH C18 column. There were good linear correlations of nine kinds of PFCAs in the range of 1.0-200.0 ng/mL, with the coefficients all more than 0.99. The limits of detection and quantification were 0.06-0.19 μg/kg and 0.19-0.62 μg/kg, respectively. The recovery rates were 70.08%-117.24% at different spiked levels ( 5.0, 25.0, 50.0 μg/kg ), and the relative standard deviations were 2.31%-19.68%. Conclusion Through optimizing the pretreatment conditions, the mobile phase of liquid chromatography and the detection conditions of mass spectrometry, the UPLC-MS/MS could meet the monitoring requirements of PFCAs in fish.

BACKGROUND:Porous gradient hydroxyapatite/zirconia composite, which can adhere to a variety of cel s, such as osteoblasts, bone marrow mesenchymal stem cel s and chondrocytes, can induce and promote fracture healing or replace bone defects. OBJECTIVE:To review the development and application of porous gradient hydroxyapatite/zirconia composite in bone tissue engineering. METHODS:Articles related to porous gradient hydroxyapatite/zirconia composite in bone tissue engineering were retrieved in CNKI and PubMed databases (1971-01/2014-12). The key words were“porous bioceramic, bone tissue engineering, bone morphogenetic protein-2, induced pluripotent stem cel s”in Chinese and English, respectively. A total of 54 articles based on inclusion criteria and exclusion criteria were obtained for the review. RESULTS AND CONCLUSION:Porous gradient hydroxyapatite/zirconia composite can provide a scaffold to induce natural y forming bone growing to fil the three-dimensional pores, thus realizing the perfect integration of tissue-engineered bone material and host-bone tissue. As zirconia functions as an enhancer of hydroxyapatite, high-quality artificial bone materials, which have elastic modulus, fracture toughness and structure similar to human bone, can be prepared by adjusting the proportion and porosity of materials. Additional y, the new hydroxyapatite/zirconia foamed ceramics as tissue-engineered bone carrying bone morphogenetic protein-2/chitosan gel sustained release system and bone marrow mesenchymal stem cel s derived from induced pluripotent stem cel s wil be expected to increase bone formation and bone fusion rates significantly in the future.

OBJECTIVETo study the expression of matrix metalloproteinase (MMP)-2, MMP-9 and vascular endothelial growth factor (VEGF)-C in breast cancer and their role in lymph node metastasis.

METHODSImmunohistochemical staining was used to detect the expression of VEGF-C, MMP-2, MMP-9 and LYVE-1 in 84 cases of breast cancer, including 52 cases with and 32 cases without lymph node metastases. The recombinant vector (pSIREN-VEGF-C) was transfected into human breast cancer cell MCF-7 by liposome, and the RNA expression of MMP-2, MMP-9 and VEGF-C in MCF-7 cells after transfection was detected by PCR.

RESULTSThe expressions of MMP-2, MMP-9 and VEGF-C were 98.1% (51/52), 88.5% (46/52), and 94.2% (49/52) respectively for the metastatic group, and 75.0% (24/32), 53.1% (17/32), and 65.6% (21/32) respectively for the non metastatic group, and there was significant difference between these groups (P < 0.05). The lymphatic vessel density between these two groups was also significantly different (P < 0.05). Increased expression of MMP-2, MMP-9 and VEGF-C was also associated with increased number of lymphatic vessels had also increased (P < 0.05). The expression of MMP-2, MMP-9 and VEGF-C in MCF-7 cells after gene transfection decreased significantly (P < 0.05).

CONCLUSIONMMP-2 and MMP-9 in conjunction with VEGF-C, promote lymphangiogenesis and lymph node metastasis of breast cancer.

Adult ; Aged ; Breast Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; surgery ; Carcinoma, Medullary ; genetics ; metabolism ; pathology ; surgery ; Cell Line, Tumor ; Female ; Humans ; Lymph Nodes ; metabolism ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Middle Aged ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor C ; genetics ; metabolism ; Vesicular Transport Proteins ; metabolism

OBJECTIVETo analyze the constituent proteins in donkey hide, the key ingredient for Ejiao, an important traditional Chinese medicine for the blood-related conditions, in hope to eventually decipher the biochemical mechanism behind Ejiao's prominent medicinal efficacy.

METHODTwo methods were employed to extract proteins in donkey skin. One used TriPure isolation reagent to extract the total proteins in donkey skin. Another used 1% sodium dodecyl sulfate (SDS) to heat the sample at 100 degrees C overnight. And then sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and capillary HPLC were used to analyze the component of proteins.

RESULTThere are not only collagen alpha1 (I) and collagen alpha2 (I), but also serum albumin in donkey skin. The content is over 25% in total proteins with the method of TriPure isolation reagent. The content of donkey serum albumin is up to 20% with the method of 1% SDS heating. And two bands, molecular weight are nearly 200 kDa,were found on 7.5% SDS-PAGE. Extracted these proteins to analyze with capillary HPLC, they were found to be the complex products of collagen and serum albumin of donkey.

CONCLUSIONDonkey serum albumin is a main protein component in the hide, which is a clue to expose is the effect of Ejiao on blood.

Animals ; Chromatography, High Pressure Liquid ; Collagen Type I ; analysis ; chemistry ; metabolism ; Collagen Type II ; analysis ; chemistry ; metabolism ; Drug Interactions ; Electrophoresis, Polyacrylamide Gel ; Equidae ; Molecular Weight ; Protein Binding ; Serum Albumin ; analysis ; chemistry ; metabolism ; Skin ; chemistry

Objective To investigate the distribution of blood vessels and the relationship of angiogenesis with hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in meningiomas.Methods Fifteen specimens of meningiomas,collected in our hospital during the resection and conformed by pathology from May 2010 to March 2011,were chosen; 4 different tissues from each specimen were taken,including the tail sign,base center,tumor' s center and tumor' s margin; three successive slices were made from each tissue part.Envision immunohistochemistry was employed to detect the expressions of CD34,VEGF and HIF-1α and their different distributions were compared.Results CD34 mainly disturbed in the cell membrane and cytoplasm,showing yellow granules in the dyeing; VEGF located at the cytoplasm of the tumor cells,while HIF-1α mainly at the nucleus.The expressions of CD34,VEGF and HIF-1α in the tail sign and tumor's margin were significantly higher than those in the MVD in the base center and tumor's center (P＜0.05).Positive correlations between expressions of VEGF and MVD,expressions of HIF-1α and MVD,as well as expressions of HIF-1α and VEGF were noted (r=0.960,P=0.040; r=0.964,P=0.036; r=0.998,P=0.002).Conclusion The angiogenesis in the periphery ofmeningiomas is much richer than that in the center; H1F-1α and VEGF may induce and promote the angiogenesis of meningiomas.

OBJECTIVETo study the content variation of furmarid acid and isofraxidin in Sarcandra glabra from 21 different provenances and provide the basis for resource utilization and quality optimization of S. glabra.

METHODHPLC method was developed to determine the contents of furmarid acid and isofraxidin in 330 samples of S. glabra which were collected respectively from 21 different provenances.

RESULTThere were significant differences in the contents of isofraxidin and furmarid acid in S. glabra from different provenances. The contents of isofraxidin and furmarid acid dropped off from low altitude to high altitude, which were also close with longitude and latitude. The content of isofraxidin in S. glabra at central area of natural distribution was the highest. The different parts of the plant had different results, the influence on the contents of the acitive components in stem were more obvious than the leaf.

CONCLUSIONThis simple, accurate and reproducible method could be use to determine the contents of furmarid acid and isofraxidin in S. glabra. The results represented the status of medicines quality and difference of Chinese S. glabra. These agreed with the traditional views that the medicines quality of Sarcandra glabra in Jiangxi, Fujian, Zhejiang was better. These provenances were considered as important areas of medicines breeding and bases building on S. glabra in future.

Chromatography, High Pressure Liquid ; Coumarins ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Fumarates ; chemistry ; Magnoliopsida ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Reproducibility of Results

AIMTo study the effect of curcumin on pulmonary arterial pressure and type I collagen of pulmonary arterioles in pulmonary hypertensive rats induced by chronic hypoxia and hypercapnia.

METHODSThirty six rats were randomly divided into three groups: normal control group (NC), hypoxic hypercapnic group (HH) and hypoxic hypercapnia + curcumin group (HC). Collagen I in pulmonary arterioles was observed by the technique of immunohistochemistry.

RESULTS(1) The findings from hemodynamics showed that the mPAP in group HH was significantly higher than that in group NC and HC. Differences of mCAP among groups were not significant (P > 0.05). (2) Light microscopy showed the value of WA/TA (vessel wall area/total area), SMC (the density of medial smooth muscle cells) and thickness of pulmonary arterial media smooth cell layer(PAMT) were significantly higher in group HH than group NC (P < 0.01) and group HC (P < 0.01). (3) Electron microscopy showed that structure of the endothelial cells in pulmonary arterioles in group HC was near to normal, and the proliferation of medial smooth muscle cells and collagen fibers in adventitia was much lighter than those of group HH. (4) Expression of collagen I in pulmonary arterioles was significantly higher in group HH than group NC (P < 0.01) and group HC (P < 0.01).

CONCLUSIONCurcumin can decrease pulmonary arterial pressure, improve pulmonary vessel remodeling and inhibit the deposition of collagen I in pulmonary arterioles.

Animals ; Arterioles ; drug effects ; metabolism ; Collagen Type I ; metabolism ; Curcumin ; pharmacology ; Extracellular Matrix ; metabolism ; Hypercapnia ; metabolism ; physiopathology ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; Male ; Pulmonary Artery ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the protective effect of N-acetylcysteine (NAC) on the intestinal barrier dysfunction in rats after extensive abdominal radiation with X ray.

METHODSTwenty-four Spraque-Dawley male rats were divided into normal control group (n=8), radiation group (n=8), and radiation+NAC group (300 mg/kg) (n=8). Radiation injury was induced by X ray with a single dose of 10 Gy. NAC was administered from 4 days before irradiation to 3 days after radiation. Three days after radiation, all the rats were euthanized. The terminal ileum was collected for crypt survival assay and ileal villi count. The tissue samples from mesenteric lymph nodes (MLN), spleen, and liver were harvested under sterile conditions for microbiological analysis and ileum samples were harvested for biochemical analysis. The blood levels of D-lactate, endotoxin and diamine oxidase (DAO) and the ileum samples levels of nitric oxide(NO) were also measured.

RESULTSRats in radiation+NAC group had a higher survival rate of intestinal crypt [(76.84+/-4.82)% vs (49.64+/-5.48)%, P<0.01], higher intestinal villus count [(8.56+/-0.68)/mm vs (4.02+/-0.54)/mm, P<0.01], lower NO concentration [(0.48+/-0.12) mumol/g vs (0.88+/-0.16) mumol/g, P<0.01], lower levels of D-lactate, endotoxin and DAO (P<0.05 or P<0.01), and significantly decreased enteric bacteria cultured from mesenteric lymph nodes and other tissues as compared with the radiation group (P<0.05 or P<0.01).

CONCLUSIONNAC protects the small intestine from radiation-induced injury maybe through the inhibition of NO in rats.

Acetylcysteine ; pharmacology ; Animals ; Dose-Response Relationship, Radiation ; Intestinal Mucosa ; drug effects ; metabolism ; microbiology ; Intestine, Small ; drug effects ; Male ; Nitric Oxide ; analysis ; Radiation Injuries ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; X-Rays ; adverse effects

ITS2 sequence was used as a barcode to identify herbal tea ingredient Plumeria rubra and its adulterants. Genomic DNAs from forty eight samples were extracted, the ITS2 sequences were amplified and sequenced bi-direstionlly, and then assembled and obtained using CodonCode Aligner. The sequences were aligned using ClustalW, the genetic distances were computed by kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic trees were constructed using MEGA5.0. Results showed that the length of ITS2 sequence of P. rubra were 244 bp. The intra-specific genetic distances (0-0. 016 6) were much smaller than inter-specific ones between P. rubra and its adulterants(0.320 8-0.650 4). The NJ tree indicated that P. rubra and its adulterants could be distinguished clearly. Therefore, Using ITS2 barcode can accurately andeffectively distinguish herbal tea ingredient P. rubra from its adulterants, which providesa new molecular method to identify P. rubra and ensure its safety in use.
Apocynaceae ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Flowers ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control

0.05)in the former part of the surgery that was before the beginning of the surgery via anterior route.But in group B,only propofol for sedation was used for the patients during the latter part of the surgery via the anterior route or while the nerve plexus was blocked; during this time in group A,the addition of fentanyl and vecuronium were still intermittently necessary to maintain the general anesthesia.The duration between the completion of surgery and the recovery of spontaneous breathing,times for initial conscious reaction such as opening the eyes following an order, extubation and from extubation to complete recovery were significantly shorter in group B than those in group A(all P