Objective:To establish a headspace GC method for the determination of 7 kinds of residual solvents in bicalutamide, including dichloromethane, n-hexane, tetrahydrofuran, ethanol, ether, acetone and ethyl acetate. Methods: The residual solvents in the substance were determined by GC equipped with an FID detector and linked with an Agilent DB-624 capillary column (30. 0 m × 0. 25 mm × 1. 4 m). The inlet temperature was 200℃ and the FID detector temperature was 250℃. The column temperature was raised by program:the initial temperature was 35℃, maintained for 13 min, raised to 180℃ with a rate of 100℃/min, and maintained for 5 min. The carrier gas was nitrogen and the flow rate was 1. 2 ml·min-1. The heated temperature of the headspace oven was 90℃, the heated time lasted 30 min, and the injection volume was 1. 0 ml. The solution medium was dimethyl sulphoxide (DMSO). Results:Each solvent could be completely separated, and the calibration curve of each solvent showed good linear relationship with good accuracy. Conclusion:The method can be applied for the determination of residual solvents in bicalutamide.

This study was aimed to investigate potential biomarkers in different traditional Chinese medicine (TCM) syndromes of angina pectoris of coronary heart disease (CHD) by using metabolomic technology,and to explore the objective law of different TCM syndromes of CHD.Endogenous metabolites in serum and urine from the healthy group,and patients with DHD angina pectoris patients of the syndrome of Qi deficiency and blood stasis,as well as the syndrome of Qi deficiency,blood stasis and phlegm turbidity were detected by the liquid chromatography-mass spectrometry (LC-MS).Metabolic profiles were analyzed by the principal component analysis (PCA) and the orthogonal partial least squares discriminant analysis (OPLS-DA).The results showed that in PCA,the healthy group,Qi deficiency and blood stasis group,as well as the Qi deficiency,blood stasis and phlegm turbidity group can be obviously distinguished.Potential biomarkers in the Qi deficiency and blood stasis syndrome contained aspartyl methionine and cysteine sulfinic acid.Potential biomarkers in the Qi deficiency,blood stasis and phlegm turbidity syndrome contained hippuric acid,amino glucose,fructosamine and triglyceride.The objective performance of Qi deficiency syndrome was the absence of biotin,lysyl tyrosine,phosphatidylglycerol and glycocholic acid.It was concluded that through the new metabolomic technology,different endogenous metabolites in the syndrome of Qi deficiency and blood stasis,as well as the syndrome of Qi deficiency,blood stasis and phlegm turbidity of patients with DHD angina pectoris were detected.It provided ideas for the clinical practice of prevention,diagnosis and treatment of different TCM syndromes.

BACKGROUND:Conditioned medium from mesenchymal stem cels (MSC-CM) may represent a promising alternative to MSCs transplantation. Previous studies have shown that inflammatory activation can strengthen the multiple biological potencies of MSCs; however, normal MSCs with insufficiency of immunocompetence and migration ability are not effective for tissue damage repair. OBJECTIVE:To investigate differential effects of MSC-CM with and without inflammatory activation on radiation-induced intestinal injury.METHODS:MSCs from the bone marrow of SD rats were separated, cultured and identified, and then co-cultured with non-irradiated IEC-6 or irradiated IEC-6 in a transwel system for 24 hours. Then, MSCs with inflammatory activation were cultured alone for another 48 hours. After that, the supernatant was colected as non-activated MSC-CM (MSC-CMNOR) and MSC-CM under radiation-induced inflammatory condition (MSC-CMIR). Rats were exposed to 14 Gy whole abdominal irradiation and randomly divided into four groups: control group, radiation injury group (DMEM/F12), MSC-CMNOR group and MSC-CMIR group. Continuous administration was givenvia tail vein and intraperitoneal implantation of Alzet microosmotic pumps. Intestinal samples were colected at 1, 3, 7 days after radiation for analysis of short circuit variation, at 3 days after radiation for analysis of intestinal epithelium ultrastructure, and at 1, 3, 5, 7, 14 days after radiation for histological observation of the intestinal epithelium using hematoxylin-eosin staining. Blood samples were colected at 1, 3, 7 days after radiation for analysis of serum xylose levels. In addition, the survival state and survival time of rats were observed and recorded. RESULTS AND CONCLUSION: The short circuit variation responding to electrical field stimulation was significantly reduced at al frequencies, but it was significantly improved in the MSC-CMIR group. Similarly, the intestinal absorption (serum xylose levels) was also significantly impaired by irradiation, but improved by delivery of MSC-CMIR (P < 0.05). At 3 days after MSC-CMIR infusion, the intestinal epithelium exhibited an increase in crypt size and vilous length (P < 0.05). Under the electron microscope, a reduction in intestinal microvili and open tight junctions in irradiated intestinal epithelium was found, and the intestine from rats treated with MSC-CMIR had more obvious tight junctions. In addition, treatment with MSC-CMIR dramaticaly improved the survival rate and mean survival time of irradiated rats as compared to those treated with DMEM/F12 or MSC-CMNOR (P < 0.05). Taken together, the present study demonstrated that MSC-CMIR , but not non-activated MSC-CM, improves the structural and functional restoration of the smal intestine after radiation-induced intestinal injury.

BACKGROUND:The establishment of a standardized clinical liver transplantation specimen bank is the primary condition for scientific research in this field, which can help to provide a qualified sample resource platform for research. OBJECTIVE:To primarily establish biological specimen bank of hepatocelular carcinoma for liver transplantation, to explore the standardized procedures of specimen colection, processing and preservation of hepatocelular carcinoma for liver transplantation, and to establish the sound and comprehensive information management system of clinical information of colected specimens. METHODS: In accordance with standardized procedures to establish biological specimen banks, the operational processes and quality control system were formulated. Liver tissue and blood samples of hepatocelular carcinoma recipients undergoing liver transplantation were regularly colected, managed and stored. Simultaneously, liver tissue and blood samples of benign liver disease in liver transplant recipients and of healthy donor were colected as controls. A systematic management was conducted in colected specimens and corresponding clinical information. RESULTS AND CONCLUSION:From August 2009, tissue and blood samples of 501 cases of receipts and donors undergoing liver transplantation with complete clinical information were colected from the specimen bank, including 203 hepatocelular carcinoma specimens, 214 benign liver disease specimens and 84 healthy donor specimens. These specimens included tumor tissue, adjacent tissues and distal non-cancerous tissue specimens, totaly 1 773. A total of 45 specimens were randomly selected for quality monitoring. The colected specimens had a high quality. Specimen information data computer management system was developed. This study initialy established a standardized research-based clinical transplantation specimen bank, which is helpful to elevate sample quality and has a good manipuility.

A rapid resolution liquid chromatography quadruple time-of-flight mass spectrometric ( RRLC-Q-TOF/MS) method combined with multivariate statistical analysis was applied to investigate the changes of endogenous metabolites in murine urine before and after ultraviolet B ( UVB ) irradiation for the purpose of discussing the physiological mechanism of acute injury caused by UVB radiation. A narrow-band UVB ( NB-UVB) (TL-01, peak value 312nm) was used to establish the acute light damage model. The urine samples were centrifuged before four times dilution treatment, subsequently the diluted urine samples were separated on a Supelco Ascentis Express C18 column using water (0. 1% formic acid) and acetonitrile as mobile phase by gradient elution. The differences metabolites with major contribution for grouping were found out based on the metabolic profiling analysis of principal component analysis ( PCA) and cluster analysis ( CA) , which could illustrate their possible mechanism of actions by means of relevant pathways. A prediction model was built to investigate the forecasting ability of the acute photo damage induced by UVB irradiation through the partial least square discriminant analysis ( PLS-DA ) . The results of multivariate statistical analysis showed that the blank control group was separated from UVB model group quite well, 11 endogenous metabolites were identified as the potential biomarkers through comparison with the database, tandem mass spectrum data and standard substance, which indicated the UVB radiation may affect the sphingolipid metabolism, nucleic acid metabolism, linoleic acid metabolism and amino acid metabolism pathways. These different metabolites could be helpful for diagnosing the light damage induced by UVB radiation

Objective To assess the relationship of methylation status of CpG islands in the promoter region of insulin-like growth factor binding protein 7 (IGFBP7) gene with the expression of IGFBP7 gene in human melanoma cell lines and primary melanocytes.Methods Primary melanocytes from human forcskin tissue as well as 4 human melanoma cell lines,including A375,M14,SK-MEL-1 and MV3,were used in this study.Bisulfite sequencing PCR (BSP) was applied to detect the methylation status of 54 CpG sites in the 5'-flanking promoter region of IGFBP7 gene in all of the melanoma cell lines and primary melanocytes.Results As hierarchical cluster analysis showed,IGFBP7-positive cells (including A375,M14 and SK-MEL-1 ) differed significantly from IGFBP7-negative cells (including MV3 cells and primary melanocytes) in the methylation pattern of IGFBP7 gene promoter region.Conclusion The methylation status of CpG island in the promoter region of IGFBP7 gene may be associated with its expression in melanoma cell lines.

Objective To investigate the association of chlamydia pneumoniae infection with ankylosing spondylitis (AS).MethodsSerum samples were obtained from 33 AS patients and 22 healthy controls.Enzyme-linked immunosorbent assay (ELISA) was applied to mearsure serum anti-Chlamydia pneumoniae antibodies (IgM/IgG),while immunofluorescence assay (IFA) was used to detect Chlamydia pneumoniae LPS antigen,and polymerase chain reaction (PCR) was used to amplify Chlamydia pneumoniae DNA in peripheral blood cells. Immunohistochemistical technique was applied to examine Chlamydia pneumoniae LPS antigen in synovial tissue from another 9 AS patients who received total hip replacement and 13 patients with comminuted femoral fractures.ResultsThe positive rates of Chlamydia pneumoniae IgM,LPS antigen and chlamydia pneumoniae DNA were higher in AS patients than those in healthy controls (78.8％ vs 22.7％,x2 =16.867,P =0.000; 66.7％ vs 31.8％,x2 =6.431,P =0.011; 33.3％ vs 9.1％,x2 =4.298,P =0.038).Chlamydia pneumoniae DNA positive rate was correlated with erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels (Z =-2.774 and -2.829,P =0.004).In synovial tissues,chlamydial LPS-containing inflammatory cells were observed in 77.8％(7/9) AS patients,while those in fracture patients was 30.8％ ( 4/13 ) ( P =0.08 ).Conclusion Chlamydia pneumoniae infection is common in blood circulation and joint cavity of AS patients and may be associated with the pathogenesis of AS.

Human cytomegalovirus (HCMV) is a DNA virus and serious opportunistic pathogen for both newborn and immunocompromised individuals.To research technique for gene silence and antiviral agents, ribozyme M1GS-T6 was constructed from external guide sequences(EGSs)that consist of a sequence complementary to HCMV UL54 gene RNA and M1 RNA, the catalytic RNA subunit of RNase P from Escherichia coli. The results showed that M1GS can efficiently cleave the mRNA sequence encoding UL54 protein in vitro.

Objective To provide theoretical and experimental proof for selecting and implying Tourette's syndrome(TS) animal models, validities of four TS models induced by chemical factors were compared. Methods Four TS models,namely AMP model,APO model,DO1 model and IDPN model were built up by using different chemical modeling agents. Through detecting spontaneous movement, climbing time and monoamine transmitters levels in striatum, four TS animal models were compared and evaluated from three levels of validities-face, prediction,construct. Results Compared with control group, spontaneous movement times raised ( t = 4. 746, P =0. 000) and level of DOPAC ( (0.99 ± 0. 177 ) ng/mg) in striatum increased (P = 0.029 ), and level of NE in striatum decreased in AMP model group( (0.11 ± 0.033 )ng/mg, P = 0.012). Compared with control group, climbing time prolonged (P = 0. 004) and levels of DA ( ( 10. 19 ± 1.23 ) ng/mg), 5-HT ( ( 0. 54 ± 0.08 ) ng/mg) in striatum raised(P=0. 019, P=0. 002),at the same time ,levels of DOPAC( (0.63 ±0.11 )ng/mg),HVA ((0.45 ±0.04 ) ng/mg) in striatum reduced (P ＜ 0.01 ) in APO model group; Compared with control group, levels of DA ( ( 13.66 ± 1.55 ) ng/mg), DOPAC( (0.80 ±0. 11 ) ng/mg), HVA( ( 1.04 ± 0.14) ng/mg) grew downwards in striatum of DOI model mice(P=0.029,P=0.001, P= 0.004). Compared with control group, level of 5-HT in striatum increased in IDPN300 group ( (0.77 ± 0.09) ng/mg, P = 0.031 ). ConclusionFace validity of AMP model is temporal and that of IDPN model is steady and persistent. AMP model,APO model and DOI model possess predictive validity. AMP model,APO model,DOI model and IDPN model have potentiality of becoming construct validity model.

Background and purpose: Increasing evidence has indicated that polymorphisms in the microRNA (miRNA, miR) binding site of target gene can alter the ability of miRNA and modulate the risk of cancer. We aimed to investigate the association between a miR-520a binding site single nucleotide polymorphism (SNP) rs141178472 in the PIK3CA 3’UTR and the risk of colorectal cancer in a Chinese Han population. Methods:The polymorphism rs141178472 was analyzed in a case-control study, including 386 colorectal cancer patients and 394 age-and sex-matched controls. The relationship between the polymorphism and the risk of colorectal cancer was examined by statistical methods. Results:Individuals carrying the rs141178472 CC genotype or C allele had an increased risk of developing colorectal cancer (CC vs TT, OR=1.716, 95%CI:1.084-2.716, P=0.022;C vs T, OR=1.258, 95%CI:1.021-1.551, P=0.033). Furthermore, the expression of PIK3CA was detected in the peripheral blood mononucleated cell of colorectal cancer patients, suggesting that mRNA levels of PIK3CA might be associated with SNP rs141178472. Conclusion:These ifndings provide evidence that a miR-520a binding site polymorphism rs141178472 in the PIK3CA 3’UTR may play crucial roles in the etiology of colorectal cancer.