AIM: To optimize the method of extracion and isolation for anthraquinone component from rhubarb for increasing the extraction rate and purity. METHODS: Rhubarb was extracted 4 times in the refluxing method with ethanol. The borate buffer solution was used as extracted fluid. Rhein, rheum emodin and chrysophanol could be obtained by Deltar 4 000 high-pressure making system, and the pure product obtained by recrystallizing with acetone. The YWG C 18 column was (4.6?250) mm, the flow rate 1.0 ml. min -1, and UV detection wavelength 254 nm. Methanol, water and acid perchloride were taken as mobile phase with the ratio of 910.01. RESULTS: Deltar 4000 high-pressure making system could be used to isolate the anthraquinone component from rhubarb, the purity of rhein, rheum emodin and chrysophanol was more than 80%, and the ertraction rate of rheum emodin reached 0.6%. CONCLUSION: The extraction of the anthraquinone component is simple and cheap with ethanol from rhubarb, and the extraction rate is high. Deltar 4000 high pressure making system can be used to isolate the anthraquinone component from rhubarb with a high the extraction rate and purity.

Chromosome Banding ; Colorectal Neoplasms ; genetics ; Comparative Genomic Hybridization ; methods ; standards ; DNA Probes ; DNA, Neoplasm ; genetics ; Humans ; Karyotyping ; Quality Control ; Reproducibility of Results

Objective To investigate the expression of matrix metalloproteinase 7(MMP7)in paraquat(PQ)?induced pulmonary fibrosis in rats. Methods Forty?eight SD rats were randomly divided into the control group and the pulmonary fibrosis model group(PQ model group),each group of twenty?four rats. Rats in the PQ model group received single intraperitoneal injection of 4 mg/mL PQ dilute solution and the control group were in?traperitoneal injected with the same dose of saline. Eight rats of each group were sacrificed on day 7,day14 and day 28 respectively. The pathological changes of lung tissues were observed and the hydroxyproline(HYP)content in lung tissues was determined. The severity of pulmonary fibrosis was observed. The expressions of MMP7 in lungs were observed by immunohistochemistry. Results The observation of general state of the experimental animals showed that except one rat died at day 28 d,all other rats survived to the end point of observation. After intraperitoneal injection with PQ,the weight of rats in the PQ model group gradually declined,and then increased around day 14,yet still much lower than that in the control group at day 28(P<0.05). After intraperitoneal injection with PQ,the pulmonary index in the model group increased gradually and then decreased after reach?ing the peak on day 14. The content of HYP in rat lung tissues in the PQ model group was remarkably higher than in the control group at day 7,day 14,and day 28,with statistical significance(P<0.01). In the PQ model group,the content of HYP was significantly up?regulated with the extension of infected time and reached the peak value at day 28. The results of HE staining showed significant pulmonary alveolitis at day 7,hyperplasia of abundant collagen fibers in alveolar septum at day 14,and obvious pulmonary fibrosis and collapse of alveolar structure on day 28 in the lung tissues of the PQ model group. A weak expression of MMP7 was measured in the lung tissues in the control group and the expression of MMP7 was higher in the PQ model group than in the control group at day 7,day 14,and day 28,with statistical significance(P<0.05). Conclusion Paraquat poison?ing was mainly manifested in inflammatory reactions of lung tissues in the early stage together with increase of fibroblasts and mainly in fibrosis in the late stage. The expression of MMP7was increased along with the severity of pulmonary alveolitis or fibrosis and showed significant changes compared to the control group at day 28 after poisoning,indicating that MMP7may be the marker of paraquat?induced pulmonary fibrosis.

To explore the risk factors of acute kidney injury(AKI)in adult primary nephrotic syndrome(PNS). Totally 185 patients with PNS were divided into AKI group(=51)and non-AKI group(=134).The demographic data and clinical and histological features at admission were compared between the two groups.The independent risk factors for AKI were evaluated by Logistics regression analysis. In 51 PNS patients with AKI,the common pathological types of AKI included minor glomerular abnormalities(29.4%),IgA nephropathy(25.5%),and membranous nephropathy(17.6%).The incidences of renal tubular casts and epithelial vacuoles in the AKI group were significantly higher than those in the non-AKI group(=0.004,=0.030).Males were more likely to suffer from AKI than females(=0.000).Patients in AKI group had significantly lower albumin level(=0.015)and higher levels of random urine protein,serum creatinine,uric acid,urea nitrogen,and triglyceride than non-AKI group(=0.030,=0.000,=0.000,=0.000,and =0.006),and polyserous and oliguria occurred more often in the AKI group(=0.000,=0.002).The AKI group had significantly higher incidences of high blood pressure and infections(=0.035,=0.000).Multivariate logistics regression analysis showed albumin(<25 g/L),serum creatinine(>96 μmol/L),urea nitrogen(≥6.8 mmol/L),uric acid(≥400 μmol/L),diabetes,infection,and renal tubular casts were the independent risk factors for AKI. AKI complicating PNS is associated with a variety of factors.Its independent risk factors include the levles of albumin,serum creatinine,urea nitrogen,and uric acid,diabetes,infections,and renal tubular casts.
Acute Kidney Injury ; etiology ; Adult ; Creatinine ; Female ; Humans ; Kidney ; Male ; Nephrotic Syndrome ; complications ; Risk Factors

Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Codon ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; DNA, Neoplasm ; genetics ; DNA-Binding Proteins ; genetics ; Exons ; Germ-Line Mutation ; Humans ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

0.05).In the followed-up patients,univariant analysis demonstrated that the expressions of CD44 and Ezrin and their coexpression were correlated with disease free survival(DFS)rate(P

Objective To construct the eukaryotic expression vectors of fibroblast growth factor receptor 3(FGFR3) MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN, and to detect their expressions in human chronic myeloid leukemia(CML)K562 cell line.Methods The full-length FGFR3 (fgfr3-WT)and dominant negative FGFR3 (fgfr3-DN)were amplified by polymerase chain reaction (PCR). The two genes were respectively digested with EcoRⅠand BamHⅠ,and then ligated into MSCV/puro to construct the recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN which were tranduced into K562 cells by LipofectaminTM 2000 after PCR,double digestion and DNA sequencing.The expressions of FGFR3 protein in K562 cells were detected by Western blotting and flow cytometry. Results The recombinant plasmids MSCV/puro-fgfr3-WT and MSCV/puro-fgfr3-DN were amplified by PCR method, and the results showed fgfr3-WT of 2 400 bp and fgfr3-DN of 1 200 bp had been successfully cloned into MSCV-puro vector. The 2 400 bp fragment was oblained after double digestion of recombinant plasmid.The sequencing results showed that the size of fgfr3-WT was 2 400 bp which was the same as the sequence from GeneBank.Fgfr3-DN of 1 200 bp was also in conformity with the expected sequence.Compared with control (K562 MSCV)group,the expression level of FGFR3-WT in MSCV/puro-fgfr3-WT transfection (K562-WT)group was increased to above 10 times.There was high expression of FGFR3-DN in MSCV/puro-FGFR3-DN transfection (K562-DN)group,but there was no expressions in control(K562 MSCV)group and K562-WT group.The flow cytometry results showed that the high expressions of FGFR3-WT were in 57.5% cells in K562-DN group and the high expressions of FGFR3-DN were in 41.5% cells in K562-DN group. Conclusion The K562 cell lines highly expressing FGFR3-WT and FGFR3-DN are constructed successfully.

Objective To explore the effect of the simvastatin administration on the expression of connective tissue growth factor （CTGF） in fihrotic lungs of rats. Methods The cats were treated with single intratracheal instillation of bleomycin （BLM） or instillation of the same volume of normal saline （NS） as a control. The administration of simvastatin（20 mg/kg）began once a day immediately or 7 days later after intratracheal BLM instillation respectively with the same volume of NS was given as a vehicle control. The rats were killed on day 7,14 and 28 respectively. Pathological alteration of lung tissue was observed hy HE staining and Masson staining. Hydroxyproline（HYP）content in lung tissue was used to determine the severity of pulmonary fibrosis. The expression of CTGF in lung tissue was exanfined by immunohistochem- istry staining and photodeusitometry. Results Histopathological changes of pulmonary fibrosis emerged gradually after the instillation of BLM. The expression level of CTGF was increased in lungs of rats after intratracheal instillation of BLM, compared with the control. The administration of simvastatin immediately or 7 days after intratracheal instillation of BLM, attenuated the histopathological changes of bleomycin- induced puhnonary fibrosis and prevented the increased expression of CTGF in lung tissue on day 28. Conclusion The adntinistration of simvastatin, immediately or 7 days after intratracheal BLM instillation, prevented the up-regulation of CTGF in fibrotic lungs of rats, which ntight be one of the mechanisms of the anti-fihrosis of simvastatin in lungs.

Background and purpose:The membrane cytoskeletal crosslinker—Ezrin may be involved in several functions including cell adhesion,motility and cell survival.There is increasing evidence that it regulates tumor progression.However,the role of Ezrin in the carcinogenesis,progression and metastasis of primary sporadic colorectal carcinoma is still under investigation.This research is to study the expression,the localization and the clinical significance of Ezrin in human sporadic colorectal carcinoma(SCRC).Methods:Immunohistological EnVision staining was used to detect the expression of Ezrin in 132 cases of human sporadic colorectal carcinoma and 43 adjacent normal colorectal mucosa,and the different expressions of Ezrin among metastatic and non-metastatic SCRC,including 74 metastatic cases and 58 non-metastatic cases were also under investigation.Results:① The expression rate of Ezrin in SCRC was significantly higher than that in adjacent normal colorectal mucosa(79.5% Vs 11.6%,P

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.
Chromatography, Liquid ; Humans ; Tandem Mass Spectrometry ; gamma-Aminobutyric Acid ; blood