The study aims to establish a quantitative nuclear magnetic resonance (QNMR) method for the determination of the absolute content of bosentan. Proton nuclear magnetic resonance spectroscopy [1H NMR] spectra were obtained in CDCl3 with the internal standard dimethyl terephthalate and zg30 pulse sequence by using a Bruker AVANCE II 400 spectrometer. The content of bosentan is determined with QNMR in comparison with the result obtained by mass balance method. The result is 96.25% by QNMR and 96.54% by mass balance method. A rapid and accurate QNMR method has been established for the quantitative determination of the absolute content of bosentan. The study provides a new way for the quality control and calibration of a new reference standard material, it could be the complementary with the mass balance method for the assay of standard reference.
Calibration ; Magnetic Resonance Spectroscopy ; methods ; Molecular Structure ; Protons ; Quality Control ; Sulfonamides ; chemistry

Objective To detect the expression of promyelocytic leukemia(PML)gene in peripheral blood lymphocytes from patients with psoriasis vulgaris,and to investigate the relationship between PML gene and psoriasis.Methods This study included 50 patients with vulgaris psoriasis who were aged between 18 and 65 years and had never received systemic treatment,as well as 45 healthy controls.Peripheral blood samples were collected from these subjects,and lymphocytes were separated by density-gradient technique.Then,RNA was extracted from the lymphocytes by using RNAprep pure blood kit followed by real time fluorescence-based quantitative reverse transcription-PCR for the detection of PML mRNA expression.Flow cytometry was performed to quantify the expression of PML protein in the lymphocytes.Independent samples t test was carried out to compare the differences in PML expression between the patients and controls.Results The relative quantity (RQ)of PML mRNA was 14.98 ± 3.64 in peripheral blood lymphocytes from the patients,significantly higher than that in the healthy controls(5.50 ± 1.10,t =16.79,P ＜ 0.05).Similarly,a significant increment was observed in the fluorescence intensity of PML protein in peripheral blood lymphocytes from the patients compared with the healthy controls(3.13 ± 0.27 vs.2.43 ± 0.21,t =6.93,P ＜ 0.05).Conclusions PML is overexpressed at the protein and mRNA levels in peripheral blood lymphocytes from patients with psoriasis vulgaris,hinting that PML may play a certain role in the development of psoriasis.

The study aims to establish a quantitative nuclear magnetic resonance (QNMR) method for the determination of the absolute content of bosentan. Proton nuclear magnetic resonance spectroscopy [1H NMR] spectra were obtained in CDCl3 with the internal standard dimethyl terephthalate and zg30 pulse sequence by using a Bruker AVANCE II 400 spectrometer. The content of bosentan is determined with QNMR in comparison with the result obtained by mass balance method. The result is 96.25% by QNMR and 96.54% by mass balance method. A rapid and accurate QNMR method has been established for the quantitative determination of the absolute content of bosentan. The study provides a new way for the quality control and calibration of a new reference standard material, it could be the complementary with the mass balance method for the assay of standard reference.

BACKGROUND:Sports-induced cartilage injury is very common; due to the poor self-healing capacity of the cartilage, cartilage repair has always been a difficult problem.
OBJECTIVE: To review the features of different seed cels in tissue-engineered cartilage construction and to explore the application of tissue-engineered cartilage construction in the repair of sports-induced cartilage injuryin vitro.
METHODS:We searched PubMed database, Wanfang database and CNKI database for articles related to tissue-engineered cartilage repair of sports-induced cartilage injuries, as wel as stem cels and scaffold materials used in tissue-engineered cartilage construction. Totaly 190 articles were retrieved, and finaly 47 articles were included in result analysis after repetitive studies were excluded.
RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cels under different conditions can differentiate into chondrocytes, and have better potential of chondrogenic differentiation compared with adipose-derived mesenchymal stem cels and umbilical cord-derived mesenchymal stem cels. But, their safety stil needs to be further studied. Good scaffolds cannot only induce stem cel differentiation, but also be the key to cartilage construction. Composite materials are the future direction of the scaffold research.

Objective To explore the effect of suramin on the epithelial-mesenchymal transition (EMT) and the excretion of transforming growth factor-β1 (TGF-β1) in peritoneal mesothelial cells (PMCs) induced by high concentrations of glucose solution (GS).Methods Cultured PMCs were divided into three groups:(1) normal control group; (2) GS-treated group:cells were treated with 1.5％,2.5％,4.25％ GS for 12 h,24 h,48 h,respectively; (3) Suramin-treated group:PMCs cultured with 4.25％ GS were exposed to different doses of suramin (25,50,100 μmol/L) for 48 h.Expression levels of α-smooth muscle actin (α-SMA) and E-cadherin were detected by Western blotting and the concentration of TGF-β1 in the culture supernatant was determined by ELISA.Results Compared with normal control group,GS-treated PMCs exhibited a time-dependent increase in the expression of α-SMA,and decrease in the expression of E-cadherin.GS also stimulated PMCs to secrete TGF-β1.In the presence of suramin,GS-induced α-SMA expression and TGF-β1 production were reduced,E-cadherin expression was increased.Conclusions Suramin can inhibit high glucose-induced EMT of PMCs by down-regulating the expression of TGF-β1.Suramin may be a novel therapeutic agent for the treatment of peritoneal fibrosis.

Background Dacryocystitis is one of the most common infectious eye diseases.The gold standard for the identification of bacteria causing dacryocystitis is bacterial culture.The combination of regular culture method with molecular biology techeniques will generate more reliable results.However,very few research data are available in ophthalmological studies in this area.Objective This study was to identify the genera and species of the dacryocystitis-causing bacteria by PCR amplification of the 16S rRNA sequences.Methods Ten cases of qualified standardized bacteria samples were taken,and the nucleic acids were released in the heating process of the PCR procedure.The 16S rRNA genes were amplified and sequenced,and the genera and species were identified using BLAST from GenBank,and the results were used to compare with the results from biochemical identification to test the reliability of this method.The cultured bacterial species from the lacrimal sac secretions from 30 cases of dacryocystitis patients were identified with the above method.Results The outcome of the PCR identification for the 10 cases of quality control standard bacterial specimens was consistent with the results from the biochemical identification.The identification of the 30 cases of dacryocystitis through sequencing the 16S rRNA revealed there were 13 cases of Staphylococcus epidermidis infection,2 cases of Staphylococcus warneri infection,1 case of Staphylococcus hominis infection,5 cases of Corynebacterium macginleyi infection,3 cases of Streptococcus pneumonia infection,2 cases of Bacillus cereus infection,1 case of Micrococcus luteus infection,1 case of Moraxella catarrhalis infection,1 case of Moraxella osloensis infection and 1 case of Pseudomonas aeruginosa infection.Conclusions Sequencing the 16S rRNA is an accurate and specific way for the identification of the genera and species of bacteria that cause dacryocystitis in patients.This sequencing method is feasible in monitoring a variety of dacryocystitis-causing pathogens.More information and epidemiological statistics about dacryocystitis can be obtained from 16S rRNA sequencing.

Background and purpose:High mobility group 1 (HMGB1), frequently found to be over-expressed in many human tumors, plays an important role in tumor progress and metastasis. This study aimed to investigate the mechanism of HMGB1 promoting A549 cell metastasis.Methods:A549 cells were untreated or treated with HMGB1 (200 ng/mL) in absence or presence of NF-κB inhibitors 6-amino-4-quinazoline (QNZ, 40 nmol/L) or Bortezomib (Bort, 20 nmol/L). Scratch assay and Transwell assay were performed to evaluate A549 cells migration and invasion ability. The activity of NF-κB was examined by luciferase reporter assay. NF-κBp65 and αvβ3 expressions were detected by Real-time RT-PCR or Western blot.Results:HMGB1 increased A549 cells migration and invasion ability. HMGB1 enhanced NF-κB protein level and NF-κB activity in A549 cells. Real-time RT-PCR and Western blot showed that HMGB1 up-regulated αvβ3 expression in A549 cells. NF-κB inhibitors QNZ or Bort reserved the promot-ing effects of HMGB1 on A549 cells migration and invasion, NF-κB expression and activity as well as αvβ3 expression. Conclusion:HMGB1 promotes A549 cell migration and invasion through activating NF-κB and up-regulating αvβ3.

Objective To explore the value of intravenous low dose furosemide on visualization of upper urinary tract during CTU. Methods 39 cases of normal upper urinary samples were examined by CTU with 5 minutes delayed,19 cases underwent intravenous injection of furosemide.The upper urinary tract was divided into 5 parts for scoring of images on a 5 score scales for opacification,the average value of ureter short axis of distention,and CT value by contrast material were measured.Results were analyzed by t test using SPSS.Results (1)30/38 segments of upper urinary tract were all or almost all opacification in furosemide group,the scores of upper urinary tract were higher than that of the control group,which had significant difference except the pelvis and left proximal ureter segments. (2)The disention of the ureter was significantly higher for all segments in furosemide group.(3)CT values in furosemide group decreased significantly for all upper urinary tract.Conclusion CTU excretory phase image acquisition with intravenous low dose furosemide is helpful on visualization of upper urinary tract.

Objective To investigate the score changes of graduates majoring in clinical medicine after using the method of “role of identification”. Methods Sixty students who were undertaking internship in Endocrinology Department of Shanghai Traditional Chinese Medicine Hospital received three-week clinical teaching and examination. Then they got into one-week “role of identification”, and were appointed as clinical teachers to teach what they have learned during the last 3 weeks to the next batch of interns and make another examination. Scores of the two examinations were compared. Results Scores of the examination after received the method in“role of identification”improved significantly (P<0.01). Conclusion The method of“role of identification”can enhance clinical medicine graduates’ mastery of professional knowledge.