Objective:To establish a GC-MS method for the simultaneous determination of 16 phthalate ester plasticizers in oral liquid preparations, compare the purification effect of liquid preparations by liquid-liquid extraction and solid phase extraction and opti-mize the operating parameters to determine the optimal experimental conditions. Methods:The samples were operated by liquid-liquid extraction and solid phase extraction, respectively. Selective ion monitor ( SIM) was adopt, phthalate esters were identified by the rela-tive abundance of major characteristic ions and the content was determined by an external standard method. Results:When the samples were operated by liquid-liquid extraction, the interference was strong with many impurity peaks. Therefore, the solid phase extraction was adopted for the sample pretreatment. GC-MS was used to detect the residues of 16 phthalate ester plasticizers in oral liquid prepara-tions. The detection limit was 0. 02μg·g-1 ,while the calibration curve showed good linearity within the range of 0. 25-8. 0μg·ml-1 with the correlation coefficient between 0. 990 7 and 0. 999 8. The average recoveries were 76. 0%-95. 4%. The relative standard devi-ations were between 2. 3% and 9. 6%(n=6). Conclusion: Pretreated by solid phase extraction, the residues of 16 phthalate ester plasticizers in oral liquid preparations can be detected by GC-MS with the properties of simple, fast, precise and sensitive, and it is suitable for the determination of phthalate esters in oral liquid preparations.

Objective To investigate the effect of ANX A1 on the biological characteristics of papillary thyroid carcinom cells by interfering with the expression of Annexina A1 (ANX A1) in papillary thyroid carcinoma cells through small interfering RNAs (siRNA).Methods The designed highly efficient siRNA was used to conduct the specific interfence on ANXA1 expression in the papillary thyroid carcinoma TPC-1 cells.The effect of ANXA1 on TPC-1 apoptosis in PTC was observed by flow cytometry.Results The designed siRAN could efficiently inhibit the expression of ANXA1 mRNA in PTC,enhanced the cell apoptosis in TPC-1 cells in vitro.Conclusion siRNA can interfere with the expression of ANXA1 and promote the apoptosis of papillary thyroid carcinoma which suggesting that ANXA1 may be an important biological target for the treatment of papillary thyroid carcinoma.

To establish a method for the determination of cucurbitacin in plasma samples, in order to study the in vivo pharmacokinetic characteristics of cucurbitacin in rats. Rats were intravenously injected with cucurbitacin. With diphenhydramine as the internal standard (IS), the plasma concentrations of cucurbitacin in rat plasma at different time points were determined by liquid chromatography tandem mass spectrometry (LC-MS/MS). With electrospray ionization source, the positive ion detection in the multiple reaction monitoring mode was conducted to determine the ion-pairs for target compound and IS were m/z 503.2/113.1 and m/z 256.0/167.2, respectively. Agilent ZOBAX SB-C18 column (2.1 mm x 50 mm, 1.8 microm) was adopted and eluted with methanol and 0.1% formic acid (55:45), and the flow rate was 0.2 mL x min(-1). DAS 2.0 software was applied to fit the blood concentration and calculate corresponding pharmacokinetic parameters. The rats were intravenously injected with cucurbitacin at the concentration of 3.0 mg x kg(-1). The target blood quality concentration show good linear relations within the range of 10.5-3 150 microg x L(-1) (R2 = 0.996), the lower limit of the standard curve was 10.5 microg x L(-1), and the signal to noise ratio S/N = 12. Intra- and inter-day precisions RSD was less than 6.9% and 14%, respectively; The accuracy RE ranged between 0.20% and 3.7%; The extraction recoveries ranged between 92.7% and 97.1%. Regarding the pharmacokinetic parameters of tail intravenous injection of cucurbitacin, AUC (0-t) was (811.615 +/- 111.578) microg x h x L(-1), (t1/2) was (1.285 +/- 1.390) h, CL was (3.627 +/- 0.487) L x h x kg(-1), and V(d) was (6.721 +/- 7.429) L x kg(-1). In this study, researchers established a simple, accurate, sensitive and highly specific method for determining the blood concentration of cucurbitacin, and reported the in vivo pharmacokinetic characteristics of cucurbitacin in rats for the first time.
Administration, Oral ; Animals ; Cucurbitaceae ; chemistry ; Cucurbitacins ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Male ; Rats ; Rats, Wistar

Objective To study the mRNA expression of HS1-associated protein X-1(Hax-1),an an- ti-apoptosis genc,in the peripheral blood mononuclear cells(PBMC)of patients with systemic lupus erythe- matosus(SLE),and further investigate the roles and significance of Hax-1 in the pathogenesis of SLE.Meth- ods Generation of longer cDNA fragments from serial analysis of gene expression(SAGE)tags for gene identi- fication(GLGI)was applied to identify the gene Hax-1 according to the Long SAGE tag.Then reverse tran- scription-polymerase chain reaction(RT-PCR)technique was used to semiquantitatively analyze mRNA ex- pressions of Hax-1 in PBMC from 34 active SLE patients and 25 healthy subjects.Results Compared with healthy controls,there was significant difference between SLE patients in the active stage and the normal controls(Z=-4.556,P＜0.01).The average level of mRNA expression in active SLE group was higher than that in healthy controls.Significant difference was found between the group with mild SLE and either the moderate or the severe one(P＜0.01).Conclusion The mRNA expression level of Hax-1 in active SLE group increase markedly,and to some extent,it is related to the activity of SLE.This provides a valuable basis for the further study on the role of apoptosis in SLE.

Objective To investigate the immortalization of human melanocytes by transfection with SV40 T antigen ( SV40T ). Methods By using SofastTM,a gene transfection reagent, the reconstructed eukaryotic expression vector SV40T-pEGFP was stably transfected into cultured primary human melanocytes, then the positive cells were selected with G418. After the positive cells were expanded in culture, the expression of SV40T gene was detected by RT-PCR and PCR, and the protein expression of SV40T by Western blotting. Results The genome DNA and total RNA were isolated from the positive cell clones, and a 288 bp fragment, which was specific for the SV40T antigen gene, was amplified. The results of immunohis-tochemistry and Western blotting confirmed the expression of SV40T protein in transfected cells. Conclusion SV40T antigen gene can successfully induce the immortalization of human melanocytes.

To compare the pharmacokinetics of syringin, eleutheroside E and isofraxidin after intravenous administration of each monomer and Ciwujia injection. Twenty-four Sprague-Dawley rats were randomly divided into four groups and intravenously administrated with syringin, eleutheroside E, isofraxidin, and Ciwujia injection, respectively. The concentrations of the three components in rat plasma were determined by LC-MS/MS. DAS 2.0 software was applied to calculate the pharmacokinetic parameters while the SPSS 17.0 software was used for statistical analysis. Significant difference (P < 0.05) was found between each monomer and the injection on the main pharmacokinetic parameters such as AUC, CL and t1,/2. Compared with the injection, the group treated with the syringin has obvious decrease in AUC, and increase in CL while the group treated with eleutheroside E has obvious increase in AUC, and decrease in CL The t1/2 of isofraxidin was prolonged in Ciwujia injection. Pharmacokinetic characters of the ingredients in the injection varied greatly from the monomer. Other constituents in the injection may have an impact on the pharmacokinetic profiles of these three components.
Administration, Intravenous ; Animals ; Coumarins ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Lignans ; administration & dosage ; blood ; pharmacokinetics ; Male ; Phenylpropionates ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

BACKGROUND:Corneal lymphangiogenesis is beneficial to the transport of corneal antigenic materials, and accelerates the process of antigen presentation, thereby playing an important role in corneal immunity. However, due to the paral el outgrowth of corneal blood and lymphatic vessels in transplanted corneas, it is often difficult to accurately evaluate the role of corneal lymphatic vessels in allograft rejection. OBJECTIVE:To explore the development of corneal lymphangiogenesis and angiogenesis in transplanted rat corneas after the blockade of vascular endothelial growth factor C (VEGF-C). METHODS:130 rats used to establish corneal al ogenic transplantation models were equally randomized into two groups:the anti-VEGF-C group and the control group. VEGF-C was blocked in the anti-VEGF-C group by intraperitoneal injection of neutralizing monoclonal anti-VEGF-C antibody every other day for 2 consecutive weeks. Meanwhile, rats in control groups received intraperitoneal injections of saline. Corneal angiogenesis and lymphangiogenesis were characterized using whole mount immunofluorescence, and the immune rejection of the grafts was evaluated by scoring the rejection index (RI). In addition, the expression of VEGF-C was examined by real-time PCR. The relationship of corneal lymphangiogenesis and angiogenesis to RI in transplanted corneas was also characterized. RESULTS AND CONCLUSION:VEGF-C expression was markedly downregulated after VEGF-C blockade. Corneal lymphangiogenesis developed in parallel with corneal angiogenesis in the control group. While there was a mild reduction in blood vessel area (BVA) and a significant decrease in lymphatic vessel area (LVA) in the anti-VEGF-C group (P<0.05). In addition, RI was positively correlated with BVA (P<0.05) and LVA (P<0.05) in the control group. However, although RI was significantly correlated with BVA (P<0.05) in the anti-VEGF-C group, the correlation between RI and LVA was not statistically significant (P>0.05). the graft survival time in the anti-VEGF-C group was significantly increased compared with that in the control group (P<0.05). Our results show that the outgrowth of lymphatic vessels is separated from that of blood vessels in transplanted corneas by blocking VEGF-C. The blockade of VEGF-C has a significant role in preventing corneal lymphangiogenesis in corneal beds, which results in higher al ograft survival rates.

Objective To investigate the clinicopathological features,diagnosis,treatment,prognosis and causative agent of a case of eumycetoma on the submaxilla.Methods A case of eumycetoma diagnosed in our department was assessed for its clinical and pathological features as well as mycologic and molecular identification.Related literature was reviewed.Results The patient was primarily characterized by swelling of the submaxilla,with multiple sinuses draining many black granules.Pathologic examination revealed a pyogenic granulomatous inflammation,and a number of lotus rhizome node-like hypha were observed in tissue samples through PAS staining.Sequence analysis of multiple loci of the isolates,including ITS 1,ITS2 and D1/D2,showed that it was mostly similar to Madurella mycetomatis with a homology of 97%.Conclusion This is a case ofeumycetoma on the submaxilla induced by a novel species of Madurella.

Data quality management system is essential to ensure accurate, complete, consistent, and reliable data collection in clinical research. This paper is devoted to various choices of data quality metrics. They are categorized by study status, e.g. study start up, conduct, and close-out. In each category, metrics for different purposes are listed according to ALCOA+ principles such us completeness, accuracy, timeliness, traceability, etc. Some general quality metrics frequently used are also introduced. This paper contains detail information as much as possible to each metric by providing definition, purpose, evaluation, referenced benchmark, and recommended targets in favor of real practice. It is important that sponsors and data management service providers establish a robust integrated clinical trial data quality management system to ensure sustainable high quality of clinical trial deliverables. It will also support enterprise level of data evaluation and bench marking the quality of data across projects, sponsors, data management service providers by using objective metrics from the real clinical trials. We hope this will be a significant input to accelerate the improvement of clinical trial data quality in the industry.
Benchmarking ; Clinical Trials as Topic ; Data Collection ; standards ; Information Storage and Retrieval ; standards ; Quality Control

Activated-Leukocyte Cell Adhesion Molecule ; Antigens, CD ; Carcinoma, Hepatocellular ; Cell Adhesion Molecules, Neuronal ; Fetal Proteins ; Hep G2 Cells ; Humans ; Liver Neoplasms ; RNA, Small Interfering