Exploring the relationships between exposure levels of bisphenols and phthalates and prostate cancer occurrence.

We established an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneously analyzing the metabolites of bisphenols and phthalates in urine to identify the associations between these exposure levels and prostate cancer (PCa) based on a case-control study. After purifying urine samples with SPE, 18 metabolites were separated on a C18 column, and MS detection was performed. The UPLC-MS/MS method has been proven effective at evaluating bisphenol and phthalate exposure (0.020-0.20 µg/L of the limits of detection, 71.8 %∼119.4 % of recoveries, 0.4 %∼8.2 % of precision). Logistic regression explored the association between exposure level and PCa in 187 PCa cases and 151 controls. The detection rates of bisphenol A (BPA) and most phthalate metabolites were 100 % ranging from 0.06-46.74 and 0.12-899.92 µg/g creatinine, respectively, while the detection rates of other bisphenols and mono-benzyl phthalate (MBzP) are low, ranging from 0 % to 21.85 %. Correlation analysis of the metabolite levels indicated that the exposure sources of BPA, di-ethyl phthalate (DEP), and di(2-ethylhexyl) phthalate (DEHP) were different, and that the exposure sources of di-n-butyl phthalate (DnBP) and di-isobutyl phthalate (DiBP) may differ between two groups. Logistic regression analysis revealed that BPA (OR<0.45 vs ≥1.43 =10.02) and DEHP exposure (OR<21.75 vs ≥45.42 =48.26) increased the risk of PCa.

Association between nickel exposure and body compositions in the United States: a population-based cross-sectional study.

BACKGROUND: Increasing body fat or decreasing muscle and bone mass were associated with worse health outcomes in the adult population. The effects of nickel exposure on body composition are not known. The aim of the current study was to investigate the relationship between urinary nickel levels and body compositions. MATERIALS AND METHODS: Two thousand seven hundred sixty-two participants were included in the analysis from the National Health and Nutrition Examination Surveys of 2017-2018 after excluding participants who have missing data on urinary nickel and those with missing all body mass component data. We used weighted generalized linear models to explore the relationship between urinary nickel and body mass components under interpolating missing covariable values. Simultaneously, sensitivity analyses and subgroup analysis were conducted to verify stability of analysis result. Curve fitting and saturation effect analysis were used to explore the possible nonlinear relationship between urine nickel and body compositions. RESULTS: Among the 2,762 participants, the average urinary nickel level was 1.58 ug/L. The weighted generalized linear models, the sensitivity analyses and subgroup analyses found no significant linear relationship between urinary nickel and body compositions. For body weight, BMI, TLM, ALM, TRF, TOF and BMC, the urine nickel saturation effect values were 0.76, 0.74, 0.5, 0.67, 0.64, 0.48, and 0.45 ug/L, respectively. For each 1 ug/L rise in urinary nickel levels at levels below the turning point, body weight increases (ß = 9.06, 95% CI = 2.75, 15.36, p = 0.01), BMI increases (ß = 3.20, 95% CI = 1.36, 5.05, p = < 0.001), TLM decreases (ß = -47.39, 95% CI = -97.38, 2.59, p = 0.06), ALM decreases (ß = -37.25, 95% CI = -63.25, -11.24, p = 0.01), TRF increases (ß = 20.68, 95% CI = 1.50, 39.86, p = 0.03), TOF increases (ß = 57.92, 95% CI = -0.12, 115.95, p = 0.05), and BMC decreases (ß = -6.84, 95% CI = -12.64, -1.04, p = 0.02). CONCLUSIONS: In summary, our study demonstrated that a dose-response relationship exists between urinary nickel and body compositions, with a low inflection point level of urinary nickel for the saturation effect.

Association between testosterone and serum soluble α-klotho in U.S. males: a cross-sectional study.

PURPOSE: Testosterone plays a crucial role in males, and the deficiency of testosterone leads to multiple adverse health conditions. Klotho is a recently discovered protein encoded by antiaging gene klotho. Both the levels of testosterone and klotho change with aging, so the relationship between them is worth exploring. The purpose of this study was to investigate whether total testosterone is associated with serum klotho levels in U.S. males aged 40-79 years. METHODS: Included in this study were 3750 male participants from the 2011 to 2016 National Health and Nutrition Examination Survey, aged 40-79 years with included information on klotho and sex hormones. The sex steroid hormone levels and klotho concentrations were assayed in laboratories using the recommended methods according to Nutrition Examination Survey guidelines. The association between sex hormones and klotho was calculated using multivariate linear regression models after adjustment for several possible confounding variables. RESULTS: Among the 3750 participants, the total testosterone concentration was 399.048 ± 184.780 ng/dL, and the testosterone deficiency prevalence was 1160 (30.942%). The geometric mean of serum klotho levels was 791.000 pg/mL. In the adjusted models, klotho increased 0.165 pg/mL for every 1 ng/dL increase of total testosterone (p = 0.004). In addition, estradiol (ß 2.232; 95% CI 0.588-3.876; p = 0.032) and sex hormone-binding globulin (ß 2.013; 95% CI 1.173-2.583; p = 0.002) were also positively associated with klotho concentrations. CONCLUSION: This study reported a significant association between klotho and sex hormones in the U.S. male population. The levels of klotho in men increased with total testosterone, estradiol and sex hormone-binding globulin levels, which may have implications for future research and clinical practice.

Apoptosis-Related Signature Predicts Prognosis and Immune Microenvironment Infiltration in Lung Adenocarcinoma.

Lung adenocarcinoma (LUAD), a malignancy with high incidence and mortality rates worldwide, contains multiple genomic and epigenomic abnormalities. And the useful tumor markers associated with these abnormalities need further investigation. Whereas apoptosis is a form of programmed cell death, the expression of apoptosis-related genes in LUAD and its relationship with prognosis is unclear. In the present study, we identified 64 differentially expressed apoptosis-related genes (DEARGs) that were differentially expressed between LUAD tissue and normal lung tissue. Based on these DEARGs, all LUAD cases were classified into two subtypes using The Cancer Genome Atlas (TCGA) cohort to assess the prognostic value of apoptosis-related genes for survival. An 11-gene signature was established by applying the Least Absolute Shrinkage and Selection Operator (LASSO) Cox regression method to construct a multigene prediction model and classify all LUAD patients in the TCGA cohort into high or low AS-score groups. Patients in the low AS-score group had significantly higher survival and prognosis than those in the high AS-score group. Taking the median risk score of the AS-score, LUAD patients in the GSE68465 cohort were divided into two risk groups, low and high. The overall survival (OS) time was longer in the low AS-score group. Combined with clinical characteristics, the AS-score was an independent predictor of LUAD patients. Gene ontology (GO) and Kyoto Encylopedia of Genes and Genomes (KEGG) analyses showed that the differential genes between the two groups were mainly enriched in cellular immunity. Further analysis revealed higher immune checkpoint protein expression and higher tumor mutational burden (TMB) in the high AS-score group, suggesting better efficacy of immunotherapy in the high AS-score group than the low AS-score group. And the high AS-score group was better in chemotherapy and targeted therapy efficiency. In conclusion, the AS-score constructed based on apoptosis-related genes can predict the prognosis of LUAD patients and provide some guidance for the antitumor treatment of LUAD patients.

Levels of urinary metabolites of benzene compounds, trichloroethylene, and polycyclic aromatic hydrocarbons and their correlations with socioeconomic, demographic, dietary factors among pregnant women in six cities of China.

Maternal exposure to benzene and related compounds, trichloroethylene, and polycyclic aromatic hydrocarbons has been suggested as risk factors for reproductive and developmental problem. In different countries and groups, there are differences in socioeconomic circumstances, consumer products, dietary habits, lifestyles, and so on, resulting in different exposure risks from these chemicals. This study investigated the correlation between their metabolite concentrations and socioeconomic, demographic, and dietary factors to explore the possible exposure source of the concerned pollutants in pregnant women. We conducted biological monitoring to assess the exposure of these chemicals using urine samples from 590 to 639 pregnant women during 2nd or 3rd trimester of pregnancy in six cities of China. Socioeconomic and demographic characteristics and dietary habits were collected from questionnaires. The detection rate was over 74% of the urine samples for all metabolites. Compared with the Fourth National Report on Human Exposure to Environmental Chemical concentrations for females (FNRHEEC, the US Centers for Disease Control and Prevention), the metabolite concentrations of benzene compounds and trichloroethylene, excluding MA reported here, were higher. Principal component analysis results showed that SBMA and MHA could be proxy of the principal sources of metabolites for benzene compounds. The concentrations of SBMA and MA were higher in Fuzhou and Wuhan, respectively. The concentration of DCVMA was higher in Shenzhen, Xi'an, and Nanning, and the concentration of PGA was higher in Fuzhou, Wuhan, and Xi'an. Also, the 1-OHPG concentration in Wuhan is higher than that in Fuzhou and Shenzhen. Unhealthy dietary habits, using cosmetics and indoor exposure, contacting chemical solvent during pregnancy were associated with increased benzene compounds, trichloroethylene, and PAH exposure. There were significant positive associations between 1-OHPG level and maternal BMI, low education status, and cooking without a range hood. Pregnant women in China may be at a greater risk of exposure to most of the target compounds than US females, and their exposure levels varied in different regions. Some adverse environmental and behavioral factors may increase the exposure of environmental toxins, which can urge people to take measures to reduce the health risk to pregnant women during pregnancy.

[Determination of Aromatic Compounds Metabolites in Human Urine by Solid Phase Extraction-liquid Chromatography-Tandem Mass Spectrometry].

OBJECTIVE: To establish the method based on high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) with solid phase extraction (SPE) for simultaneous determination of the biological metabolites of aromatic compounds, including N-acetyl-S-phenyl-L-cysteine (SPMA), N-acetyl-S-benzyl-cysteine (SBMA), p-nitrophenol (PNP), methylhippuric acids (MHA), p-Aminophenol (PAP), mandelic acid (MA), phenylglyoxylic acid (PGA) and 1-hydroxypyrene (1-OHP) in urine. METHODS: After adding 20 µL of ß-glucuronidase and 1 mL ammonium acetate buffer solution in 1 mL of urine, the sample was digested in a 37 ℃ incubator for 20 h. After digestion, the enzymatic hydrolysate was purified by PRIME HLB solid phase extraction column. The target compounds were eluted with 4 mL of acetonitrile and blown to dryness with nitrogen, reconstituted with 0.20 mL of methanol. Injected the sample solution into LC-MS/MS system for analysis after filtering with 0.22 µm filter membrane. LC separation was carried out on a reversed-phase C18 column (2.1 mm×150 mm, 3.5 µm); gradient eluting was performed at a flow rate of 0.2 mL/min. The water containing 0.1% formic acid was used as mobile phase A and methanol was used as mobile phase B. The mass spectrometry was performed with multiple reaction monitoring (MRM) mode, using alternating positive and negative ions, and internal standard curves were used for quantification. RESULTS: The eight metabolites showed good linearity within the range of 1-100 ng/mL, with a correlation coefficients greater than 0.995, and the relative precision deviation (RSDs) was 0.050%-9.95%. The method detection limits (MDLs) of the eight target metabolites were 0.041-0.12 ng/mL. The proposed method was used for urine sample analysis and the spiked recoveries were 80.1%-114.0%. CONCLUSION: The established method is quick, sensitive and accurate; it meets the requirementof the biological monitoring of aromatic compounds for the general population and occupational population.

[Determination of 8-oxo-2'-deoxyguanosine and Cotinine in Urine by Hydrophilic Chromatography-tandem Mass Spectrometry with Isotope Dilution].

OBJECTIVE: To develop an assay for determination of 8-oxo-2'-deoxyguanosine and cotinine in human urine by hydrophilic chromatography tandem mass spectrometry (HILIC-MS/MS) with isotope dilution. METHODS: The urine supernatant was 1∶5 diluted with 3 mmol/L ammonium formate aqueous solution containing 15N 5-8-OHdG and D 3-cotinine as internal standard. After being filtered through a 0.22 µm water filter, the sample solution was injected into ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. Separation was performed on ACQUITY UPLC® BEH HILIC column (50 mm×3.0 mm, 1.7 µm) with isocratic elution (A∶B=10∶90) at 40 ℃. The mobile phase was composed with acetonitrile (B) and 3 mmol/L ammonium formate water soulution (A). The flow rate was 0.3 mL/min. Positive ion scan-multiple reaction monitoring (MRM) mode were used for monitoring and internal standard curves were applied for quantification. RESULTS: Good linearity was obtained under the optimal conditions. Detection limits for 8-OHdG and cotinine were 0.064 µg/L and 0.035 µg/L respectively, the quantitation limits were 0.21 µg/L and 0.12 µg/L respectively, and the recoveries of the spiked urine samples were 92.6%-102% and 102%-106% respectively. Statistical analysis of 40 urine sample determination results obtained by using the above assay showed that there were significant differences in tobacco smoke exposure and tobacco-specific nitrosamine intake between active and passive smoker ( P<0.05). The concentration of NNAL and cotinine were higher in urine samples of active smoker. Tobacco smoke exposure was positively correlated with tobacco specific nitrosamine intake in both active and passive smokers (the correlation coefficients were 0.487 and 0.786 respectively, P<0.05). CONCLUSION: We successfully established a simple and fast assay for simultaneously detecting 8-oxo-2'-deoxyguanosine and cotinine in human urine. It was sensitive and accurate for quntification via the calibration by the isotope internal standards, and can meet the needs of batch analysis.

Investigating isomers/enantiomers of perfluorooctanoic acid in river water by gas chromatography-mass spectrometry with chiral derivatization.

Perfluorooctanoic acid (PFOA) in environmental media contains numerous isomers/enantiomers because of the PFOA manufacturing process and biological degradation of PFOA precursors. Few methods for analyzing PFOA enantiomers have been described. A simple derivatization method using (S)-1-phenethyl chloride that was developed to allow PFOA isomers/enantiomers to be separated by gas chromatography and analyzed by electron-capture negative ionization mass spectrometry is described here. PFOA standards were analyzed, and enantiomers of the chiral isomers perfluoro-3-methyl-heptanoic acid, perfluoro-4-methyl-heptanoic acid, and perfluoro-3,5-dimethyl-hexanoic acid were separated using an HP-5MS column. Linear PFOA and perfluoro-6-methyl-heptanoic acid were chromatographically separated from these enantiomers. The linear ranges (giving correlation coefficients r > 0.997) of the calibration curves for the isomers were 0.010-3.00 ng/mL. PFOA isomer/enantiomer concentrations in river water were determined using the method. The method separated the enantiomers of perfluoro-3-methyl-heptanoic acid and perfluoro-4-methyl-heptanoic acid, the isomers of perfluoro-6-methyl-heptanoic acid, and linear PFOA in river water. No significant differences were found between the PFOA enantiomer/isomer compositions of the sample and technical PFOA. Enantiomer ratios can provide information about the sources and transport of pollutant isomers/enantiomers in the environment. Enantiomeric separation requires effective separation techniques. Our method achieved chiral separation using a non-chiral GC column that is often used in general analytical laboratories. The method could be used to investigate the sources and fates of PFOA and the isomers/enantiomers of other potentially toxic persistent pollutants in the environment and the risks posed to humans.

[Detecting Nicotine and Cortinine in Hair by Hydrophilic Interaction Liquid Chromatography Tandem Mass Spectrometry].

OBJECTIVE: To develop a method for detecting nicotine and cotinine in hair by hydrophilic interaction chromatography tandem mass spectrometry. METHODS: Hair samples were hydrolyzed in sodium hydroxide solution before extraction with dichloromethane. The samples were blown to dry with nitrogen and dissolved with mobile phase. The filtrate of the samples was injected into a chromatographic-mass spectrometry system for analysis. The separation was performed by a hydrophilic column, with which methanol-0.1% ammonia was used as the mobile phase. The quantitative detection of Nicotine and Cortinine was carried out with electron spray ionization-triple quadrupole mass spectrometry. The established method was used for detecting nicotine and cotinine in 602 hair samples of pregnant women and 31 hair and urine samples of volunteers. RESULTS: A standard curve was drawn for the established method of hydrophilic liquid chromatography tandem mass spectrometry. Good linearity was obtained for detecting nicotine and cotinine in the range of 0.030-100.000 µg/L, with a detection limit (MDL) of 0.007 6 µg/g and 0.004 4 µg/g, respectively. The inter-day and intra-day precisions reached a level of less than 10%. The recoveries of the spiked samples ranged from 81.0% to 102.0%. About 0.020-0.260 µg/g nicotine and 0.004 8-0.069 0 µg/g cotinine were detected in the pregnant women without exposure to secondhand smoking (SHS), compared with 0.029-0.350 µg/g nicotine and 0.005 6-0.085 0 µg/g cotinine in those exposed to SHS. Nicotine and cotinine were also found in the hair and urine samples of volunteers, which were correlated with smoking (P < 0.05). A dose-response relationship were found between smoking and hair nicotine. CONCLUSIONS: The proposed method is accurate and sensitive for detecting nicotine and cotinine in hair samples. Hair nicotine can be a specific biomarker for assessing exposure to tobacco smoking.

Application of response surface methodology to vortex-assisted dispersive liquid-liquid extraction for the determination of nicotine and cotinine in urine by gas chromatography-tandem mass spectrometry.

A method of vortex-assisted dispersive liquid-liquid extraction coupled with gas chromatography and tandem mass spectrometry for the determination of nicotine and cotinine in urine was developed. Response surface methodology was applied to obtain the optimum extraction conditions. In this method, Plackett-Burman design was utilized to evaluate the impact of five selected factors on pretreatment procedure. Then, three main factors were optimized using a Box-Behnken design. The optimized method showed good linearities at 1-2000 µg/L with correlation coefficients of 0.9998 for nicotine and 0.9986 for cotinine. Recovery was 91.4-106 and 91.7-108% for nicotine and cotinine, respectively. The intraday relative standard derivations of determination were 1.47-4.06% for nicotine and 0.41-3.16% for cotinine, and interday relative standard derivations were 3.03-6.70% for nicotine and 1.64-6.38% for cotinine. The method detection limits for nicotine and cotinine were 0.33 and 0.34 µg/L, respectively. A total of 87 urine samples from smokers and nonsmokers were tested with the proposed method. Urinary nicotine and cotinine were 23.0-6.67 × 103 and 18.4-4.17 × 103  µg/(g·cr) for smokers and 1.31-286 and 1.39-131 µg/(g·cr) for nonsmokers, respectively. The method is sensitive, suitable and reliable for the determination of nicotine and cotinine in urine and meets the requirements for evaluating short-term tobacco exposure.

[Detection of 11 Organophosphate Pesticides and Atrazine in Water by Dispersive Liquid-Liquid Microextraction Combined with Gas Chromatography Mass Spectrometry].

OBJECTIVE: To develop a method for detecting 11 organophosphate pesticides and atrazine in water samples by dispersive liquidliquid microextraction combined with gas chromatographymass spectrometry (DLLMEGC/MS) . METHODS: DLLME and GCMS parameters were optimized for efficient extraction of chemicals. The proposed method was used for detecting organophosphate pesticides in tap water and river water samples,with 200 µL of dimethylbenzene as extractant and 800 µL of methanol as dispersant. They were mixed,emulsified and dispersed into 10 mL of water samples. The extractant (1 µL) from centrifugation was injected into the GC/MS system for analyses. GC separation was performed on HP5 column (30 m×0.25 mm,0.25 µm) by temperature programming. Mass spectrometric analysis was done with EI and selected ion monitoring (SIM) mode was used for quantitative analysis. RESULTS: Good linear ranges for detecting the 11 pesticides and atrazine appeared from 2.0 ng/mL to 50 ng/mL,with a limit of 0.121.38 ng/mL. The relative standard derivations (RSDs) ranged from 5.57% to 9.85%. The average recoveries ranged from 75.5% to 107%. CONCLUSION: The method is sensitive and rapid,with simultaneous extraction and concentration procedures. The lowdensity organic solvent after extraction is easy to isolate. The method fits for analyses of organophosphate pesticides and atrazine in water samples.

[Determination of nicotine and cotinine in urine by hydrophilic interaction chromatography-tandem mass spectrometry].

The analytical method of nicotine and cotinine in human urine with hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS/MS) was established. After the urine sample containing nicotine-d4 and cotinine-d3 isotope internal standards being diluted with water, the filtrate was introduced into ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for analysis. Separation was performed on an ACQUITY UPLC® BEH HILIC column (50 mm×3.0 mm, 1.7 µ m), in which methanol and 0.1% (v/v) ammonia were used as the mobile phases with isocratic elution at 0.2 mL/min of flow rate. Positive ion scan mode was used for mass spectrometry measurement and calibration curves were plotted for quantification determination. A good linearity could be obtained in the range of 1.0-1000 µ g/L for nicotine and cotinine with the linear coefficients of 0.9949 and 0.9958, respectively. The limits of detection of nicotine and cotinine were 0.082 µ g/L and 0.077 µ g/L, and the limits of quantification were 0.27 µ g/L and 0.26 µ g/L, respectively. The recoveries of the spiked urine samples were 90.4%-103.5% and 93.0%-104.6%, and the relative standard deviations (RSDs) were 4.80%-6.21% and 4.22%-7.15% for nicotine and cotinine respectively. The established method was applied to the analyis of 200 urine samples. Based on the investigation information of the urine of the smoking people, the nicotine contents were 26.68-854.30 µ g/L, and the cotinine contents were 36.66-1191.18 µ g/L (n=86, Mnicotine=76.00 µ g/L, Mcotinine=83.52 µ g/L, M:median); of the nonsmoking people, the nicotine contents were 5.08-69.66 µ g/L, and the cotinine contents were 3.16-28.21 µ g/L (n=114, Mnicotine=7.53 µ g/L, Mcotinine=3.79 µ g/L). The method is simple, sensitive and rapid. It is suitable for batch analysis of nicotine and cotinine in urine, and it can meet the requirement of evaluating the human tobacco exposure.

Bone marrow mesenchymal stem cells repair cadmium-induced rat testis injury by inhibiting mitochondrial apoptosis.

Cadmium is a highly toxic metal with widespread exposure to people that can cause tissue injuries that lack effective treatment. The aim of this project was to uncover whether bone marrow mesenchymal stem cells (BMSCs) can repair cadmium-induced rat testis injury and to explore the role of mitochondrial apoptosis in this process. To this end, 21 adult male Wistar rats were randomly divided into control, model and therapy groups, 7 each, and were administered 0, 0.4 and 0.4 mg/kg body weight CdCl2 saline solution, respectively, by intraperitoneal injection 5 times per week for 5 weeks. Then, rats in the therapy group were treated with 107 BMSCs by retro-orbital injections, while the others were given equal volumes of phosphate buffered saline. Following 2-week BMSCs-treatment, the therapy rats were heavier than the model rats, despite there being no difference in testicular cadmium contents between these groups, which were both significantly higher than the control group. BMSCs were observed in the testis of the therapy rats, in which pathological changes improved significantly compared with the model group. Expression of the apoptosis-associated proteins Bim, Bax, Cytochrome C, Caspase-3, active-Caspase-3 and AIF increased, while Bcl-2 was reduced significantly in rat testes of model group compared with the other groups. Based on these findings, we conclude that cadmium can accumulate in rat testes where it caused severe tissue injury, BMSCs can be localized to the injured testicular tissue of rats and repair the tissue injury, these reparative effects may be highly related with mitochondrial apoptosis.

A simple analytical method of determining 1-hydroxypyrene glucuronide in human urine by isotope dilution with ultra performance liquid chromatography-tandem mass spectrometry.

The glucuronide conjugate of 1-hydroxypyrene (1-OHP-G) is a sensitive and reliable index biomarker for assessing low exposure to polycyclic aromatic hydrocarbons (PAHs). A simple method for determining 1-OHP-G in human urine with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established and applied to evaluate the exposure level of PAHs of pregnant women in a large sample size. After the urine sample was extracted with ethyl acetate, 0.2 mL of the aqueous phase was diluted to 1.0 mL with 5 mmol/L ammonium acetate before injection. The chromatographic separation was performed on a C18 column with a gradient elution and identification was conducted on a tandem mass spectrometry with electrospray ionization in negative mode. 1-OHP-d9-G was used as an internal standard to improve precision. The method was validated and good linearity was obtained in the range of 0.1∼2.0 ng/mL. The limit of detection (LOD) and the limit of quantification (LOQ) of 1-OHP-G were 0.015 and 0.051 ng/mL. Intra-day and inter-day precision were 4.3 and 6.7 %, respectively. The spiked recoveries were 79.4∼106 % for urine samples. This method was rapid, sensitive, and very suitable for batch analysis of urine. Six hundred seventy-five urine samples of pregnant women from the cities of Fuzhou, Shenzhen, and Nanning of P.R. China were analyzed with the proposed method. The medians of 1-OHP-G concentration were 0.27 µg/g.cr (n = 201), 0.30 µg/g.cr (n = 212), and 0.51 µg/g.cr (n = 262) for the cities of Fuzhou, Shenzhen, and Nanning, respectively. 1-OHP-G concentrations in urine samples of pregnant women from the cities of Fuzhou and Shenzhen in coastal areas were both significantly lower than that of Nanning City in inland region (p < 0.001). Graphical Abstract The internal standard 1-OHP-d9-G and 2.0 mL of ethyl acetate were added to 1.0 mL of urine sample, after vortex vibration and centrifugation the aqueous phase was removed and diluted, and 5 µl of aliquot was analyzed by UPLC-MS/MS.

[Determination of Nicotine and Cotinine in Urine by Dispersive Liquid-Liquid Microextraction Combined with Gas Chromatography-Mass Spectrometry].

OBJECTIVE: To develop a method for the determination of nicotine and cotinine in urine samples by dispersive liquid-liquid microextraction (DLLME) combined with gas chromatography-mass spectrometry (DLLME-GC/MS). METHODS: The experimental conditions for GC-MS and DLLME were investigated in detail. DLLME was performed with the following procedure: 5 mL of urine sample was adjusted to pH9. 0 with NaOH solution; NaCl was added to increase ionic strength; 100 µL chloroform (containing internal standard of quinolone) as extractant was mixed with 1 000 pL methanol as dispersant and then injected into the urine sample to make it emulsified and dispersed. The sample solution was centrifuged for 5 min at 4,000 r/min, and 1 µL of its extraction solvent was injected into the GC/MS system for analysis. GC separation was performed with DB-5 column under programmed temperature. Nicotine and cotinine were quantified using selected ion monitoring (SIM) mode of mass spectrum detection and internal standard working curve. RESULTS: Good linear relationship was obtained for detecting nicotine and cotinine ranging from 0. 2 ng/mL to 100 ng/mL, with a detection limit of 0. 010 ng/mL and 0. 022 ng/mL for nicotine and cotinine respectively. The relative standard derivation (RSD) for determination of nicotine and cotinine in urine samples were 8.2% and 9.6% respectively. The spiked recoveries ranged from 92.0% to 108.0% for nicotine, 83.0% to 110.0% for cotinine. CONCLUSION: The method is rapid, sensitive, accurate and simple, with little consumption of organic solvent. It is suitable for determination of nicotine and cotinine in urine, and can meet the requirements for evaluating human tobacco exposure.

[Effectiveness of CLAT Protocol for Treating Patients with Refractory Acute Myeloid Leukemia].

OBJECTIVE: To explore the clinical efficacy and toxicity of CLAT protocol (cladribine, cytarabine and topotecan) for treating patients with refractory acute myeloid leukemia (R-AML). METHODS: A total of 18 patients with R-AML (median age 37 years, range 18 to 58 years; male n = 16, female n = 2) were treated with CLAT protocol, which consisted of cladribine 5 mg/m(2)/d, i.v. on days 1-5, cytarabine 1.5 g/m(2)/d, i.v. on days 1-5, topotecan 1.25 mg/m(2)/d, i.v. on days 1-5 and G-CSF 300 µg/d subcutaneous injection on day 6 until neutrophile granulocyte recovery. RESULTS: Out of 18 patients 2 died of severe infection before the assessment. Among 16 evaluated patients, 10 (55.6%) achieved complete remission (CR), and 2 (11.1%) achieved partial remission (PR), the overall response rate was 66.7%, the rest 4 patients did not respond (NR). The median overall survival time and DFS for the CR patients was 9.5 months (95%CI: 6.7-16.64) and 9.5 months (95%CI: 6.1-16.7) respectively. The 1 year OS and DFS rates were 45% and 46.9%, respectively. All patients developed grade 4 of granulocytopenia and thrombocytopenia, the median duration was 13 (range 2 to 21) days and 12 days (range 2 to 21), respectively, all patients developed infection, 2 patients died of severe infection. The most common non-hematological side effects included nausea, vomiting, diarrhoea, rash, aminotransferase or bilirubin elevation and were grade 1 to 2. CONCLUSION: The CLAT protocol seems to have promising for the treatment of refractory AML patients, and patients well tolerated. This CLAT protocol offers an alternative treatment for R-AML patients who received severe intensive treatment, especially with anthracycline-containing chemotherapy.

Lichong decoction reduces Matrix Metalloproteinases-2 expression but increases Tissue Inhibitors of Matrix Metalloproteinases-2 expression in a rat model of uterine leiomyoma.

OBJECTIVE: To study the effect of Lichong decoction (LD) on expression of matrix metalloproteinase- 2 (MMP-2) and metalloproteinase-2 (TIMP-2) in a rat model of uterine leiomyoma (UL). METHODS: UL was induced in rats using exogenous estrogen and progesterone. LD was administered (p.o.) for 4 weeks, and mifepristone (RU-486) used as a control. To observe the effect of LD on the uterine coefficient and uterine transverse diameter, a radioimmunoassay method was used to detect serum levels of sex hormones. Light microscopic analyses of pathologic changes in the tissues of UL rats were evaluated. Expression of the proteins of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in uterine tissues was assessed by immunohistochemical staining and western blotting. RESULTS: A UL model in rats was established successfully. LD reduced uterine weight, uterine coefficient, and uterine transverse diameter compared with untreated controls. LD reduced levels of estradiol, progesterone, follicle-stimulating hormone, and luteinizing hormone in our UL models. LD improved the pathologic condition of uterine muscle. Expression of MMP-2 protein decreased to varying extents in LD-treated groups, but TIMP-2 levels were enhanced. LD appears to reduce MMP-2 expression and increase TIMP-2 expression in UL tissue. CONCLUSION: These data suggest that the mechanism of action of LD on ULs may involve reduction of MMP-2 expression and increase in TIMP-2 expression in rats.

Association of increased circulating catecholamine and glucocorticoid levels with risk of psychological problems in oral neoplasm patients.

BACKGROUND: Noradrenergic pathways and glucocorticoid-mediated signal pathways have been implicated in the growth and progression of oral cancer. Patients with oral neoplasms can have high psychological distress levels, but the effects of stress-related hormones on oral neoplasm growth are unknown. METHODS: We have investigated the relationships between pre-surgical measurements of psychological problems with Symptom Checklist-90-revised Inventory (SCL90-R), tumor histology, circulating blood catecholamine and glucocorticoid levels among 75 oral neoplasm patients, including 40 oral cancer patients and 35 benign oral tumor patients. RESULTS: The results showed that most dimension scores of SCL90-R did not show a significant difference between the two groups except depression (p = 0.0201) and obsessive-compulsion (p = 0.0093), with the scores for these symptoms being higher among oral cancer group versus the benign oral tumor group. The differences of total score, average score and other monomial factor scores were not statistically significant. The mean concentrations of catecholamine and glucocorticoid in peripheral blood of the oral cancer group were higher than those in benign oral tumor group (p<0.01). We also examined whether associations observed between biobehavioral measures and circulating blood catecholamine and glucocorticoid levels extended to other compartments in the oral cancer group. CONCLUSIONS: These findings suggest that stress hormones may affect oral cancer behavior by influencing the tumor micro-environment though the circulating blood.

[Determination of organochlorine pesticides in water using solidification of floating organic drop liquid phase microextraction by gas chromatography].

OBJECTIVE: To develop a new method for simultaneous determination of eight organochlorine pesticides (OCPs) in water samples using solidification floating organic drop liquid-phase microextraction (SFO-LPME) combined with gas chromatography-electronic capture detector (GC-ECD). METHODS: The experimental conditions of SFO-LPME were determined with n-Hexadecane as extractant, water samples adjusted to pH 6.0, inonic strength increased by adding 15.0 g/100 mL NaCI. The OCPs were extracted at 55 degrees C for 10 mm and determined with GC-ECD. RESULTS: A good linearity (correlation coefficients > or = 0.996) was obtained for eight target compounds from 5 ng/I. to 100 ng/L. The method detection limits ranged from 0.24 ng/L to 0.78 ng/L. Satisfactory results were achieved with samples of river water, piped water and farmland water, with an average recovery of 76.0%-106.0% and RSD of 3.24%-11.60%. CONCLUSION: The proposed method is rapid, simple and sensitive. It is suitable for hatch analyses of eight organic chlorine pesticides in water samples.

[Preliminary study of assessment of mental health level among oral tumor patients and difference of relevant material basis].

OBJECTIVE: To investigate the difference in mental health status of oral tumor patients and their spouses, and explore the differences on the basis of relevant materials. METHODS: Forty patients with oral cancer, eighteen spouses, and thirty-five patients with oral benign tumor were diagnosed in the West China Hospital of Stomatology between December 2011 and August 2012 and assessed with symptom checklist-90 (SCL90) (the 5-grade scoring). Participants were assessed independently according to their conditions. Blood samples were obtained from the participants by syringe on the second admission day. A method was developed to determine the concentrations of catecholamine and glucocorticoid in the serum using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). RESULTS: The scores for compel, depression and rests in the cancer group and benign tumor group were statistically significant (P<0.05). The scores for compel, dread and stubborn in the cancer group and their spouses were statistically significant (P<0.05). The differences between the total and other monomial factor scores were not statistically significant. However, the contents of epinephrine, norepinephrine, cortisone and hydrocortisone in the serum, as determined by HPLC-MS/MS, were significantly different (P<0.001). CONCLUSION: Psychiatric factors do not show a simple factor effect on patients. Symptoms of patients may be based on tumorigenesis and developed in small molecules. Further research is required.