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

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.

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.

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.

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 analyze the clinical manifestations and electroencephalogram (EEG)characteristics of agyria-pachygyria for its early diagnosis,treatment and prognosis judgment in clinical practice.Methods The clinical manifestations and EEG features of twenty-four patients with agyria-pachygyria who were diagnosed by CT or magnetic resonance imaging(MRI) at Pediatric Neurology of Children's Hospital of Chongqing Medical University from July 2004 to July 2013 were retrospectively analyzed.Results Of twenty-four patients,eighteen cases were diagnosed as diffuse agyria-pachygyria and six cases were diagnosed as partial agyria-pachygyria.The clinical features were mainly manifested as mental retardation (twenty-four patients),and motor retardation (twenty-four patients),and epilepsy (eighteen patients).All of the twenty-four patients had abnormal EEG pattern which were mainly three tapes.Type Ⅰ had diffused high amplitude alpha and beta activity in all cortical regions,frontal-central,or parietal-occipital region (fourteen patients).Type Ⅱ showed alternating high amplitude bursts with sharp and slow waves (seven patients).Type Ⅲ was characterized by high amplitude spike or sharp wave activity generalized or multifocal distribution and δ,θ wave mixing graphics (twelve patients).Nine of twenty-four patients showed two or three EEG characteristic patterns in an awake-asleep EEG recording.During the follow-up of 1-8 years old,twelve of the thirteen patients who were diagnosed as epilepsy in diffuse agyria-pachygyria had refractory epilepsy,mainly with infantile spasms or Lennox-Gastaut syndrome.One of the five patients who was diagnosed as epilepsy in focal agyria-pachygyria had refractory epilepsy,mainly for partial epilepsy secondary generalized seizures.There was a significant difference between them (P =0.008).Eighteen of twenty patients who had moderate-severe mental retardation or dyskinesia were diagnosed as diffuse a gyria-pachygyria,while two were focal agyria-pachygyria.Both of them had a significant difference (P =0.005).Conclusions Agyria-pachygyria is a brain malformation caused by neuronal migration abnormality.Diffuse agyria-pachygyria is presented with serious clinical manifestations and poor outcome while the clinical manifestation of focal agyria-pachygyria is relatively mild and epilepsy could be controlled by antiepileptic drugs or epilepsy surgery.These characteristics of EEG patterns along with clinical findings could provide important evidence for early diagnosis,timely treatment and prognosis judgment of agyria-pachygyria.

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.

Objective To investigate the prevalence of and risk factors for neurosyphilis in syphilitic patients with persistently positive nontreponemal serological tests after regular therapy.Methods Clinical data were collected from 248 syphilitic patients with persistently positive nontreponemal serological tests after regular therapy,and analyzed retrospectively.Univariate analysis,multivariate logistic regression analysis and receiver operating characteristic (ROC) curve analysis were performed to assess factors associated with neurosyphilis.Results Of the 248 patients,25 (10.1％) were diagnosed with neurosyphilis.As univariate analysis showed,the occurrence of neurosyphilis was associated with the degree of decline in serum toluidine red unheated serum test (TRUST) titers (x2 =20.663,P ＜ 0.05) and persistent positive serum TRUST titers (Z =-7.021,P ＜ 0.05),but not with gender,age,stage of syphilis,treatment protocols,initial serum TRUST titers,or the presence or obsence of neurological symptoms (P ＞ 0.05).Multivariate logistic regression analysis revealed that persistent positive serum TRUST titers were a risk factor for neurosyphilis (OR =4.685,95％ CI =2.552-8.601,P ＜ 0.05).ROC curve analysis showed that the best cut-off point of persistent positive serum TRUST titer was 1 ∶ 8 for predicting neurosyphilis,with the area under the curve (AUC) being 0.907.Conclusion Serum TRUST titers are somewhat valuable for predicting neurosyphilis in syphilitic patients after regular therapy.

Objective To investigate diagnostic magnetic resonance imaging features of orbital Langerhans cell histocytosis (LCH)and improve its diagnostic accuracy.Methods The symptoms and image data of fourteen histophathology verified orbital LCH cases are reviewed and analyzed.Results Nine patients had swollen eyelids,accompanying with symptoms of inflammation, esphthalmos and orbital masses.One case had cough symptom and another had diabetes insipidus.Of these fourteen cases,seven occurred in right orbital,six occurred in left orbital and one involved bilateral orbital.As to the location of LCH,six cases located in super-lateral wall of the orbit,five cases located in lateral wall of the orbital,and three cases located in roof of orbital.On MRI, thirteen cases lesions show hypo or iso signal intensity on T1 WI,and eleven cases lesions show heterogeneous hyper or iso signal in-tensity on T2 WI.The lesions of eosinophilic granuloma has clear border,which differentiate it from other types.After contrast en-hancement,MR imaging showed marked inhomogeneous enhancement.Conclusion MRI is the primary modality in diagnosing of or-bital LCH,clearly and accurately manifesting the extent of orbital LCH.It will be helpful to diagnose LCH timely if combining with clinical data.MR could provide reliable information for making surgical operation and treatment plan.