1. Simultaneous detection of five kinds of olefine chlorides in workplace air by solvent desorption-gas chromatography
Siyan LI ; Yuanqin ZHONG ; Lihe YE
China Occupational Medicine 2018;45(02):223-226
OBJECTIVE: To establish a method for simultaneous detection of five kinds of olefine chlorides in workplace air by solvent desorption-gas chromatography. METHODS: The olefine chlorides in workplace air were collected using activated charcoal tubes,desorbed by carbon disulfide,separated on a HP-INNOWAX capillary column and detected with flame ionization detector. RESULTS: The linear range of the five kinds of chlorinated olefins( vinyl chloride,allyl chloride,chloroprene,trans-1, 2-dichloroethylene, cis-1, 2-dichloroethene) was wide and the linear correlation coefficient was greater than 0. 999 8. The detection limit and the minimum detectable concentrations were 0. 30-0. 80 mg/L and 0. 05-0. 13 mg/m~3,respectively( air sample volume 6. 0 L). The recovery rate was 95. 90%-102. 67%. The within-run and betweenrun relative standard deviations were 0. 00%-3. 74% and 0. 21%-6. 12%,respectively. The samples could be preserved at room temperature for at least 14 days. CONCLUSION: This method has simple operation,good linear range,high sensitivity and good reproducibility,which is suitable for detecting 5 kinds of olefine chlorides in workplace air.
2. Detecting human urinary thiocyanate by gas chromatography with pre-column derivatization
Xiaozhou ZHUANG ; Xiaolin RUAN ; Weifeng RONG ; Jianmei PENG ; Lihe YE ; Xiaoxuan CAI ; Fenghua ZHANG ; Yuanqin ZHONG
China Occupational Medicine 2018;45(02):219-222
OBJECTIVE: To establish a method for detecting human urinary thiocyanate by gas chromatographic and pre-column derivatization with 2,3,4,5,6-pentafluorobenzyl bromide( PFB-Br). METHODS: A total of 20. 0 μL of urine was taken and 1. 0 m L of acetonitrile and 100. 0 μL of PFB-Br were added for derivative reaction. The gas chromatography was directly used for measurement. RESULTS: The urinary thiocyanate concentration showed a good linear range of 1. 000-10. 000 mg/L. The linear correlation coefficient was 0. 999 6. The minimum detection concentration was 0. 112 mg/L,and the minimum quantitative concentration was 0. 411 mg/L( 20. 0 μL urine sample). The standard recovery rate was 97. 22%-102. 04%.The within-run relative standard deviation( RSD) of this method was 1. 56%-5. 35%. The between-run RSD was 1. 46%-5. 10%. Hydrocyanic acid ions interfered with the measurement. Other common inorganic ions such as chloride,sulfate,and nitrate ions did not interfere with the measurement results. The samples can be stored at 4 ℃ for at least 15 days. CONCLUSION: This method is suitable for detecting human urinary thiocyanate.
3.Effect of Biejia Decoction Pill on aerobic glycolysis in hepatocellular carcinoma by regulating the protein kinase B/mammalian target of rapamycin signaling pathway
Qinwen TAN ; Jingjing HUANG ; Ruixi ZHONG ; Yuanqin DU ; Jian XU ; Jinli NONG ; Yujiao PENG
Journal of Clinical Hepatology 2025;41(2):300-306
ObjectiveTo investigate the inhibitory effect of Biejia Decoction Pill on the proliferation, migration, and aerobic glycolysis of hepatocellular carcinoma (HCC) using cell experiments, as well as related mechanisms. MethodsHuman liver cancer cell line Huh7 was selected, and Sprague-Dawley rats were randomly divided into blank serum group, inhibitor group, and high-, middle-, and low-dose Biejia Decoction Pill groups. Rat serum containing the drug was prepared for the incubation of Huh7 cells. CCK8 assay and scratch assay were used to explore the effect of Biejia Decoction Pill on the proliferation and migration of HCC cells; glycolytic rate-limiting enzymes and metabolites were measured to explore the effect of Biejia Decoction Pill on aerobic glycolysis of liver cancer cells; RT-qPCR and Western blot were used to explore the effect of Biejia Decoction Pill on the mRNA expression, related proteins, and phosphorylation of the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test or the Dunnett’s T3 test were used for further comparison between two groups. ResultsCompared with the blank serum group, the Biejia Decoction Pill groups had significant reductions in OD value, migration rate during different periods of time, glycolytic rate-limiting enzymes (hexokinase, phosphofructokinase, pyruvate kinase), and glycolytic metabolites (pyruvate, lactic acid, ATP) (all P<0.05). RT-qPCR results showed that compared with the blank serum group, the high-, middle-, and low-dose Biejia Decoction Pill groups had a significant reduction in the mRNA expression level of mTOR, and the high- and low-dose Biejia Decoction Pill groups had a significant reduction in the mRNA expression level of AKT (all P<0.05). Western blot results showed that compared with the blank serum group, the high-, middle-, and low-dose Biejia Decoction Pill groups had significant reductions in the expression levels of mTOR-related proteins and phosphorylated proteins, and the high- and middle-dose Biejia Decoction Pill groups had significant reductions in the expression levels of AKT-related proteins and phosphorylated proteins (all P<0.05). ConclusionThis study preliminarily verifies that the serum containing Bijia Decoction Pill can inhibit the aerobic glycolysis of human hepatoma Huh7 cells, thereby inhibiting their proliferation and migration, possibly by inhibiting the expression of the proteins related to the AKT/mTOR signaling pathway.
4.Therapeutic effect of retention enema with compound rhubarb decoction on a rat model of minimal hepatic encephalopathy based on bile acid metabolomics
Yuanqin DU ; Meng WANG ; Guochu HUANG ; Chun YAO ; Ruixi ZHONG ; Liangjiang HUANG ; Jian XU ; Jingjing HUANG ; Qinwen TAN ; Dewen MAO
Journal of Clinical Hepatology 2023;39(10):2348-2357
ObjectiveTo investigate the therapeutic effect of rhubarb decoction (RD) retention enema on a rat model of minimal hepatic encephalopathy (MHE) and its mechanism of action based on bile acid (BA) metabolomics. MethodsA total of 55 male Sprague-Dawley rats were randomly divided into blank group (NC group with 10 rats), hepatic encephalopathy group (HE group with 15 rats), MHE group with 15 rats, and MHE+rhubarb decoction treatment group (MHEY group with 15 rats). Intraperitoneal injection of carbon tetrachloride (CCl4) and thioacetamide (TAA) was performed to establish a rat model of MHE or HE, and the rats were sacrificed after 2 weeks of administration. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin (TBil), and total bile acid (TBA) and the concentration of blood ammonia were measured; the colonic contents were collected to measure pH value; liver and brain tissue samples were collected, and HE staining was used to observe the histopathological changes of the liver; the bile was collected, and liquid chromatography-mass spectrometry was used to perform BA-targeted metabolomics analysis. Continuous data were expressed as mean±standard deviation; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the NC group, the HE group and the MHE group had a significant increase in searching platform latency (after modelling and after administration) and a significant reduction in the number of platform crossings (all P<0.05); compared with the MHE group, the MHEY group had a significant reduction in searching platform latency (after administration) and a significant increase in the number of platform crossings, and the HE group had a significant increase in searching platform latency and a significant reduction in the number of platform crossings (all P<0.05). Compared with the NC group, the HE group and the MHE group had significant increases in AST, ALT, ALP, TBil, TBA, blood ammonia, and colon pH value (all P<0.05); compared with the MHE group, the MHEY group had significant reductions in AST, ALT, ALP, TBil, TBA, blood ammonia, and colon pH value (all P<0.05), and the HE group had significant increases in AST, ALT, ALP, TBil, TBA, blood ammonia, and colon pH value (all P<0.05). The MHE group had significantly lower TBA, primary BA, and secondary BA than the NC group (all P<0.05); compared with the MHE group, the HE group had significantly lower TBA and primary BA (all P<0.05), and the MHEY group had significantly higher TBA and primary BA (all P<0.05). Compared with the NC group, the MHE group had significant reductions in GCDCA, GUDCA, GHDCA, TCDCA, TUDCA, GLCA, and TLCA (all P<0.05) and significant increases in γ-MCA, THCA, 7-KDCA, AlloLCA, and α-MCA (all P<0.05), and compared with the MHE group, the MHEY group had significant increases in THDCA, TMCA, TCDCA, TUDCA, and TLCA (all P<0.05). ConclusionRD retention enema can improve liver injury and cognitive function in a rat model of MHE induced by CCl4 and TAA by regulating the enterohepatic circulation of BA, possibly by increasing the synthesis of taurine-binding BA.
5. Simultaneous determination of 14 chlorinated hydrocarbons in urine by headspace gas chromatography-mass spectrometry
Xiaoxuan CAI ; Si TANG ; Jianmei PENG ; Xiaozhou ZHUANG ; Lihe YE ; Siyan LI ; Yuanqin ZHONG ; Zheng MA ; Huipeng CHEN ; Fenghua ZHANG
China Occupational Medicine 2020;47(04):470-474
OBJECTIVE: To establish a headspace gas chromatography-mass spectrometry method for the determination of 14 chlorinated hydrocarbons in urine. METHODS: The urine sample 4.00 mL and anhydrous sodium sulfate 3.00 g were added into a 10.00 mL headspace bottle, then the headspace bottle was maintained at 70 ℃ for 40.0 min. After headspace pretreatment, 14 chlorinated hydrocarbons in headspace air were separated in the DB-5 MS capillary column of the gas chromatography and detected by mass spectrometer. RESULTS: There was a good linear relationship of 14 chlorinated hydrocarbons in urine in the range of 0.62-1 630.00 μg/L. The linear correlation coefficient was greater than 0.999 0.The minimum detectable concentration was 0.19-0.43 μg/L and the minimum quantitative concentration was 0.62-1.44 μg/L. The average recovery rate was 89.8%-107.1%. The within-run relative standard deviation(RSD) was 4.0%-8.5% and the between-run RSD was 6.3%-9.1%. Urine samples can be stored at 4 ℃ or-8 ℃ for 3 days and below-20 ℃ for 7 days. CONCLUSION: This method is rapid, simple, sensitive, accurate and has little interference,which can be used as a method for detecting 14 kinds of chlorinated hydrocarbons in urine samples of patients with occupational poisoning.