{L-End}Objective To establish a method for the simultaneous determination of dimethyltin (DMT), trimethyltin (TMT), diethyltin (DET), and triethyltin (TET) in human whole blood using high performance liquid chromatography-inductively coupled plasma-mass spectrometry (ICP-MS). {L-End}Methods The 1.0 mL of blood was added with 4.0 mL 65% aqueous solution (containing 6% acetic acid), extracted and separated by C₄ column (150 mm×3 mm×3 μm) using a mobile phase of methanol and 4% acetic acid aqueous solution (containing 0.25 mmol/L tropolone) at a volume ratio of 35∶65, and detected by ICP-MS. {L-End}Results The linear range of DMT, TMT, DET, and TET was 30.60-550.80, 29.00-522.00, 46.10-829.80, and 34.05-612.90 μg/L, respectively. All correlation coefficients were 0.999. The detection limit of DMT, TMT, DET and TET was 21.40, 20.30, 32.27 and 23.80 μg/L, respectively. The recovery rate was 81.9%-104.9%. The within-run and between-run relative standard deviation was 1.6%-6.9% and 0.1%-10.0%, respectively. The samples can be stored at -20 ℃ and 4 ℃ for at least three days. {L-End}Conclusion This method can be used for trace analysis of DMT, TMT, DET, and TET in whole blood.

OBJECTIVE: To explore the feasibility of simultaneous detection of 45 kinds of common organic compounds in workplace air by solvent desorption-gas chromatography method.METHODS: A total of 45 kinds of common organic compounds such as benzene,1,2-dichloroeyhane,n-hexane and trichloroethylene in workplace air were collected with activated carbon tube and desorbed with carbon disulfide,separated by capillary chromatographic column,and detected with flame ionization detector.RESULTS: There was good linear relationship in the selected range.The correlation coefficients was 0.999 92-0.999 99.The detection limit was 0.03-0.30 mg/L and the minimum detectable concentration range was 0.01-0.20 mg/m~3( sample volume was 3.00 L).The average desorption efficiencies was 75.4%-105.7%.The within-run and between-run relative standard deviations were 0.4%-6.7% and 1.8%-7.9%,respectively.The sampling efficiency was 91.9%-100.0%.CONCLUSION: The method is simple,high sensitivity and good precision,which can be used for simultaneous detection of 45 kinds of common coexisting organic compounds in workplace air.

Filamin A and 14-3-3-σ are closely associated with the development of breast cancer.However,the exact relationship between them is still unknown.The present study aimed to examine the interaction of filamin A with 14-3-3-σ in the invasion and migration of breast cancer.RNA interference technology was employed to silence filamin A in MDA-MB-231 cells.Real-time PCR and Western blotting were used to detect the expression of filamin A and 14-3-3-σ at mRNA and protein levels,respectively.Double immunofluorescence was applied to show their colocalization morphologically.Wound healing assay and Trans-well assay were used to testify the migration and invasion of MDA-MB-231 cells in filamin A-silenced cells.The results showed that silencing filamin A significantly increased the mRNA and protein levels of 14-3-3σ.In addition,double immunofluorescence displayed that filamin A and 14-3-3σ were predominantly colocalized in the cytoplasm of MDA-MB-231 cells.Silencing filamin A led to the enhanced fluorescence of 14-3-3σ.Furthermore,cell functional experiments showed that silencing filamin A inhibited the migration and invasion of MDA-MB-231 cells in vitro.In conclusion,silencing filamin A may inhibit the invasion and migration of breast cancer cells by upregulating 14-3-3σ.

OBJECTIVE: To establish a methodology for simultaneous detection of chloromethyl methyl ether( CMME) and bis-chloromethyl ether( BCME) in workplace air by gas chromatography. METHODS: CMME and BCME in workplace air were collected with absorption solution which was also derivatization solution. The derivative products were extracted using n-hexane alkaline medium. The extracts were separated by capillary column and detected with electron capture detector.The quantification was performed by use of standard curves. RESULTS: The linearity ranges of CMME and BCME were2. 00-80. 00 and 1. 32-52. 80 ng,respectively. The correlation coefficients were both 0. 999 93. The minimum detectable concentrations were both 0. 030 μg / m3 and the minimum quantification concentrations were both 0. 100 μg / m3( 7. 50 L sample). The recovery rates were 99. 35%-101. 00% and 97. 99%-101. 70% respectively. The within-run relative standard deviations( RSD) were 2. 73%-4. 46% and 2. 61%-3. 82% respectively,and the between-run RSD were3. 10%-5. 50% and 3. 89%-5. 38% respectively. The sampling efficiencies were 92. 43%-96. 25% and 91. 43%-94. 03%respectively. The samples were stable at room temperature for at least 15 days. CONCLUSION: This method is suitable for simultaneous detection of CMME and BCME in workplace air.

Objective To establish pretreatment conditions of hippuric acid (HA) and methyl-hippuric acid (MHA) in urine and HPLC conditions.Methods HA and MHA in urine were extracted with acetonitrile under acid condition and determinated by HPLC-DAD.The operating conditions by HPLC were C18 column (150 mm× 4.6 mm,5 μm),methanol-0.2％ acetic acid (contained 6.5 mmol/L potassium dihydrogen phosphate) (25:75,V/V) as mobile phase,1 ml/min as flow rate and wavelength was at 254 nm.Results The standard curves for HA,2-MHA and 3-MHA (4-MHA) showed good linearity between 9.91～2 974.20 μg/ml (r=0.999 98),1.91～ 573.60 μg/ml (r=0.999 84) and 2.00～598.65 μg/ml (r=0.999 85),respectively.The mean recoveries were 96.38％～98.01％,83.17％～94.05％,103.22％～104.45％,respectively.The within-run precision were 0.50％～ 1.20％,0.51％～1.59％,0.49％～0.95％,respectively,and the between-run precision were 1.70％～3.20％,1.30％～ 2.67％,0.86％～2.74％,respectively.The detection limit of HA,2-MHA and 3-MHA (4-MHA) were 0.18 μg/ml,0.46 μg/ml and 0.12 μg/ml,and the low determination concentrations of the method were 0.36 μg/ml,0.92 μg/ml and 0.24 μg/ml (1 ml urine).The urine can be kept 15 days at 4 ℃ refrigerator without significantly loss.Conclusion This method with simply pretreatment conforms to the relevant requirements of guide for establishing occupational health standards-part 5:determination methods of chemicals in biological materials.It can be used to detect HA and MHA in urine for occupational population exposure to toluene and xylene.

Objective To establish pretreatment conditions of hippuric acid (HA) and methyl-hippuric acid (MHA) in urine and HPLC conditions.Methods HA and MHA in urine were extracted with acetonitrile under acid condition and determinated by HPLC-DAD.The operating conditions by HPLC were C18 column (150 mm× 4.6 mm,5 μm),methanol-0.2％ acetic acid (contained 6.5 mmol/L potassium dihydrogen phosphate) (25:75,V/V) as mobile phase,1 ml/min as flow rate and wavelength was at 254 nm.Results The standard curves for HA,2-MHA and 3-MHA (4-MHA) showed good linearity between 9.91～2 974.20 μg/ml (r=0.999 98),1.91～ 573.60 μg/ml (r=0.999 84) and 2.00～598.65 μg/ml (r=0.999 85),respectively.The mean recoveries were 96.38％～98.01％,83.17％～94.05％,103.22％～104.45％,respectively.The within-run precision were 0.50％～ 1.20％,0.51％～1.59％,0.49％～0.95％,respectively,and the between-run precision were 1.70％～3.20％,1.30％～ 2.67％,0.86％～2.74％,respectively.The detection limit of HA,2-MHA and 3-MHA (4-MHA) were 0.18 μg/ml,0.46 μg/ml and 0.12 μg/ml,and the low determination concentrations of the method were 0.36 μg/ml,0.92 μg/ml and 0.24 μg/ml (1 ml urine).The urine can be kept 15 days at 4 ℃ refrigerator without significantly loss.Conclusion This method with simply pretreatment conforms to the relevant requirements of guide for establishing occupational health standards-part 5:determination methods of chemicals in biological materials.It can be used to detect HA and MHA in urine for occupational population exposure to toluene and xylene.

Forkhead Box c2 (FOXC2) is a member of forkhead/winged-helix family of transcription factors. The relationship between FOXC2 and invasive breast cancers, including basal-like breast cancer (BLBC, a subtype of breast cancer), remains to be elucidated. In this study, immunohistochemistry was used to detect the expression of FOXC2 in samples from 103 cases of invasive breast cancers and 15 cases of normal mammary glands. The relationship between FOXC2 and clinical parameters of invasive breast cancers such as patient's age, tumor size, lymph node metastasis, tumor grade, the expression of ER, PR, HER-2 and p53, and Ki-67 labeling index (LI) was evaluated. The expression of FOXC2 was detected in parent MCF7 cells, MCF cells transfected with FOXC2 expression vectors and MDA-MB-435 cells by immunohistochemistry and Western blotting. Transwell assay was used to determine the invasive ability of these cells. The results showed that FOXC2 was strongly expressed in basal epithelial cells in normal mammary glands and weakly expressed or even not expressed in glandular epithelial cells. The majority of invasive breast cancers (71.8%, 74/103) had negative or weak expression of FOXC2. However, FOXC2 was strongly expressed in 60.7% of BLBCs. Moreover, FOXC2 was related with tumor grade, p53 expression, ki-67 LI and lymph nodes metastasis. It was expressed in FOXC2-transfected MCF cells and MDA-MB-435 cells but not in parent MCF cells. Transwell assay revealed that MCF cells transfected with FOXC2 expression vectors were more aggressive than the parent MCF cells, suggesting a positive correlation between FOXC2 and the invasion of breast cancer. It was concluded that there is a significant association between FOXC2 and the metastasis of invasive breast cancer. FOXC2 may be used as a new marker for the diagnosis and prognosis prediction of different subtypes of invasive breast cancers.
Biomarkers, Tumor ; biosynthesis ; Breast Neoplasms ; diagnosis ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Forkhead Transcription Factors ; biosynthesis ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Neoplasm Proteins ; biosynthesis ; Prognosis

Breast Neoplasms ; classification ; genetics ; metabolism ; pathology ; Chromosome Aberrations ; Female ; Gene Deletion ; Humans ; Phyllodes Tumor ; classification ; genetics ; metabolism ; pathology ; Twist-Related Protein 1 ; metabolism ; Wnt Signaling Pathway

OBJECTIVETo detect the expression of phosphorylated-signal transducer and activator of transcription 3 (p-Stat3) and myeloid leukemia-1 (Mcl-1) as well as their correlation, and to investigate the functional role of Stat3 and Mcl-1 in the pathogenesis of esophageal squamous cell carcinoma (ESCC).

METHODSStat3 activity in ESCC cells was inhibited with JAK/Stat3 inhibitors (AG490 or JSI-124). Specific siRNA was used to inhibit the Stat3 expression. Cell apoptosis was detected by flow cytometry. Expression of Mcl-1 protein was determined by Western blotting. Expression of phospho-Stat3 (Tyr705) and myeloid leukemia-1 (Mcl-1) proteins in ESCC tissues was detected by tissue microarray and immunohistochemistry. The relationship between p-Stat3 or Mcl-1 aberrant expression and clinicopatholohical features of ESCC was analyzed. The correlation of their expression was also analyzed.

RESULTSSuppression of the Stat3 signaling activation in ESCC cells led to marked apoptosis, and dramatic reduction of Mcl-1 protein. The positive rate of phospho-Stat3 (Tyr705) expression was 45.0% in 50/111 of the ESCC tissue samples. The lower the degree of tumor differentiation, the higher the positive rate of phospho-Stat3 (Tyr705), showing a significant difference (P = 0.018). The positive rate of Mcl-1 protein expression was 72.1% (80/111), and the lower the degree of tumor differentiation was, the higher there was the positive rate of Mcl-1, with a significant difference (P = 0.026). There was a positive correlation between the expressions of p-Stat3 and Mcl-1 proteins (P = 0.012).

CONCLUSIONSIn a subset of ESCC tissues, p-Stat3 (Tyr705) and Mcl-1 are overexpressed and positively correlated with each other, and both are correlated with tumor differentiation. Persistent activation of Stat3 contributes to apoptotic resistance in ESCC cells, and may be at least partly mediated through upregulation of Mcl-1.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cell Survival ; drug effects ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Neoplasm Grading ; Neoplasm Staging ; Phosphorylation ; RNA, Small Interfering ; genetics ; STAT3 Transcription Factor ; antagonists & inhibitors ; genetics ; metabolism ; Tyrphostins ; pharmacology

Animals ; Cell Differentiation ; Cell Movement ; Chemokine CXCL12 ; metabolism ; Endothelial Cells ; pathology ; physiology ; Humans ; Muscle, Smooth, Vascular ; pathology ; physiology ; Receptors, CXCR4 ; metabolism ; Stem Cells ; metabolism ; pathology ; physiology ; Vascular Diseases ; metabolism ; pathology ; physiopathology