1.Effect of Marsdenia tenacissima combined with XELOX solution on disulfide apoptosis in human colorectal cancer HCT116 cells
Wei WEI ; Zhaoying CAI ; Yayun QIAN
Journal of International Oncology 2024;51(9):545-555
Objective:To investigate the effect and related mechanism of Marsdenia tenacissima combined with XELOX solution on disulfide apoptosis in human colorectal cancer HCT116 cells.Methods:The human colorectal cancer HCT116 cells were cultured in vitro and treated with different concentrations of capecitabine (0, 0.3, 0.6, 1.2, 2.4, 4.8 μg/ml), oxaliplatin (0, 10, 20, 40, 80, 160 μg/ml), Marsdenia tenacissima (0, 15, 30, 60, 120, 240 mg/ml), and the glucose inhibitor BAY-876 (0, 4, 8, 16, 32, 64 μg/ml), respectively. Furthermore, the HCT116 cells were pre-treated with 25 μg/ml of BAY-876, followed by exposure to the specified concentrations of each drug group. HCT116 cells were divided into the following groups: negative control group (no treatment), capecitabine group (4.0 μg/ml), oxaliplatin group (90 μg/ml), Marsdenia tenacissima group (140 mg/ml), XELOX solution group (4.0 μg/ml capecitabine+90 μg/ml oxaliplatin), and Marsdenia tenacissima combined with XELOX solution group (140 mg/ml Marsdenia tenacissima+4.0 μg/ml capecitabine+90 μg/ml oxaliplatin). BAY-876 treatment groups refer to the groups which the glucose inhibitor BAY-876 25 μg/ml was added to each of the above groups. The cell proliferation was assessed using the MTT assay. Apoptosis was determined through Annexin Ⅴ-FITC/PI double staining. The concentration of glucose was quantified using the o-toluidine method. NADPH levels were measured by colorimetry. Cystine uptake fluorescence assay was utilized to quantify the fluorescence intensity of cystine, and cysteine content was determined using cysteine colorimetry. Results:After 0, 0.3, 0.6, 1.2, 2.4, 4.8 μg/ml concentration of capecitabine, 0, 10, 20, 40, 80, 160 μg/ml concentration of oxaliplatin, 0, 4, 8, 16, 32, 64 μg/ml concentration of BAY-876, and after pre-treatment with 25 μg/ml glucose inhibitor BAY-876, HCT116 cells were treated with the above drugs and concentrations, there were statistically significant differences in cell survival rate ( F=644.60, P<0.001; F=417.30, P<0.001; F=1 028.00, P<0.001; F=1 066.00, P<0.001; F=847.70, P<0.001), and with the increase of each drug concentration, the activity of HCT116 cells decreased gradually (all P<0.05). After 0, 15, 30, 60, 120, 240 mg/ml concentration of Marsdenia tenacissima, and after pre-treatment with 25 μg/ml glucose inhibitor BAY-876, HCT116 cells were treated with the above drugs, there were statistically significant differences in cell survival rate ( F=107.50, P<0.001; F=619.70, P<0.001), and with the increase of drug concentration, the activity of HCT116 cells increased first and then decreased (all P<0.05). Annexin Ⅴ-FITC/PI staining showed that the intensity of green, red and yellow fluorescence was weaker in the negative control group. The expression of green fluorescence and red fluorescence was enhanced in each drug group. In capecitabine group, the red fluorescence and yellow fluorescence were larger. The proportion of green fluorescence in oxaliplatin group and Marsdenia tenacissima group was smaller. The proportion of green fluorescence and yellow fluorescence in XELOX solution group was higher than that in capecitabine group and oxaliplatin group. Marsdenia tenacissima combined with XELOX solution group had the highest proportion of green and red fluorescence. After pre-treatment with the glucose inhibitor BAY-876, the green and yellow fluorescence of the cells in each group increased significantly. The green and yellow fluorescence of capecitabine group and oxaliplatin group increased significantly. The fluorescence of XELOX solution group was significantly higher than that of capecitabine group and oxaliplatin group. In Marsdenia tenacissima combined with XELOX solution group, the proportion of fluorescence expressing three colors was the largest. In the negative control group, capecitabine group, oxaliplatin group, Marsdenia tenacissima group, XELOX solution group, Marsdenia tenacissima combined with XELOX solution group, and the groups after BAY-876 pre-treatment, the glucose concentration of HCT116 was (19.91±0.13), (22.82±0.88), (11.87±0.14), (17.93±0.14), (10.53±0.10), (7.56±0.08), (11.44±0.10), (11.73±0.72), (8.98±0.40), (14.25±0.33), (6.77±1.50), and (1.56±0.17) μg/ml, respectively, with statistically significant differences ( F=762.60, P<0.001; F=118.80, P<0.001). Compared with the untreated groups, the intracellular glucose concentration of HCT116 cells treated with BAY-876 was significantly decreased ( t=86.50, P<0.001; t=16.90, P<0.001; t=11.83, P<0.001; t=17.79, P<0.001; t=4.35, P=0.012; t=54.34, P<0.001). NADPH levels in each group were (131.80±2.61), (93.87±1.00), (136.50±3.69), (105.70±0.84), (146.90±2.94), (105.00±2.25), (92.33±0.23), (88.63±0.31), (97.33±2.02), (81.77±1.33), (102.80±1.61), and (85.13±0.45) nmol/gProt, respectively, with statistically significant differences ( F=225.60, P<0.001; F=125.50, P<0.001) ; Compared with the untreated groups, the intracellular NADPH levels of HCT116 cells treated with BAY-876 was significantly decreased ( t=26.11, P<0.001; t=8.62, P<0.001; t=16.13, P<0.001; t=26.38, P<0.001; t=22.78, P<0.001; t=14.97, P<0.001). The fluorescence intensity of cystine in each group was 607.30±8.76, 655.70±6.57, 647.10±19.35, 737.80±6.34, 756.00±8.65, 846.60±11.70, 929.60±6.88, 1 049.00±22.35, 1 021.00±29.49, 1 094.00±16.17, 1 137.00±10.08, and 1 230.00±46.57, respectively, with statistically significant differences ( F=188.00, P<0.001; F=48.32, P<0.001). Compared with the untreated groups, the intracellular fluorescence intensity of cystine of HCT116 cells was significantly increased after treatment with BAY-876 ( t=50.09, P<0.001; t=29.26, P<0.001; t=18.34, P<0.001; t=35.53, P<0.001; t=49.66, P<0.001; t=13.83, P<0.001). The contents of cysteine in each group were (457.00±30.69), (581.20±30.69), (326.40±5.49), (374.20±5.54), (565.30±5.54), (246.80±30.69), (100.30±16.57), (472.90±19.10), (262.70±28.65), (348.70±9.55), (533.40±11.03), (30.23±5.49) μmol/L, respectively, with statistically significant differences ( F=110.00, P<0.001; F=423.50, P<0.001). Compared with the untreated groups, the intracellular contents of cysteine of HCT116 cells treated with BAY-876 was significantly decreased ( t=17.71, P<0.001; t=5.19, P=0.006; t=3.78, P=0.019; t=4.00, P=0.016; t=4.47, P=0.011; t=12.03, P<0.001) . Conclusion:Marsdenia tenacissima combined with XELOX solution can promote HCT116 cell death through disulfide apoptosis.
2.Investigation on knowledge and practice of iodine deficiency disorders prevention and control among main caregivers of pregnant women in trimester
Liangliang HUO ; Zhaoying QIAN ; Bing ZHU ; Zhou SUN ; Tingting ZHAO ; Shuchang CHEN ; Qingxin KONG
Chinese Journal of Endemiology 2022;41(7):561-565
Objective:To learn about the knowledge and practice of iodine deficiency disorders (IDD) prevention and control among main caregivers of pregnant women in trimester and its related influencing factors, so as to provide a scientific basis for effectively improving the iodine nutrition level of pregnant women.Methods:In April 2020, using the stratified sampling method, 2 districts and counties were selected in the urban, suburban and rural areas of Hangzhou City, respectively, and 5 maternal and child health care institutions were selected as the survey sites in each district and county according to the five directions of East, West, South, North and Middle. A face-to-face questionnaire investigation was used to investigate the knowledge and practice of IDD prevention and control among main caregivers of pregnant women.Results:Totally 601 caregivers of pregnant women were investigated. Among them, 68.9% (414/601) knew that eating iodized salt was the best way to prevent IDD. However, 15.0% (90/601) thought that eating seafood was the best way to prevent IDD. And 41.9% (252/601) caregivers of pregnant women would actively purchase iodine-rich food for cooking, and 63.6% (382/601) would choose iodized salt. There were statistically significant differences in the knowledge and practice of IDD prevention and control among caregivers of pregnant women in urban, suburban and rural areas ( P < 0.05). And 50.3% (87/173) male caregivers would actively purchase iodine-rich food for cooking, which was higher than that of female caregivers [38.6% (165/428)], the difference was statistically significant (χ 2 = 6.97, P = 0.008). There was a statistically significant difference between the caregivers of pregnant women of different ages in actively purchasing iodine-rich food for cooking (χ 2 = 12.61, P = 0.013). And there were statistically significant differences between the caregivers of pregnant women with different educational levels in actively purchasing iodine-rich food for cooking and choosing iodized salt (χ 2 = 29.34, 39.42, P < 0.001). Conclusions:The awareness of knowledge about IDD prevention and control and the formation of prevention and control behavior among caregivers of pregnant women in Hangzhou City are poor. Active education of caregivers of pregnant women should be strengthened to promote maternal and infant health.
3.Correlation between the Detection of Multiple Cytokine Levels in Neonatal Umbilical Cord Plasma and Early-onset Neonatal Sepsis
Zhengjiang CHANG ; Qian WEI ; Xiaolong LIU ; Zhaoying FU ; Meini CHEN ; Jinwei HE ; Haiyan FAN
Journal of Modern Laboratory Medicine 2024;39(4):150-154
Objective To investigate correlation between the expression level of multiple cytokine levels in neonatal umbilical cord plasma and early-onset sepsis for screening out the cytokines with good diagnostic value for early-onset neonatal sepsis(EONS).Methods Full-term neonates and preterm neonates(Gestational age ≥ 32 weeks)of 310 cases between September 2021 and June 2023 were selected as study subjects.According to clinical signs,laboratory results and blood culture,these subjects were divided into 3 groups:control group without sepsis,EONS blood culture positive group and EONS blood culture negative group.Umbilical cord blood plasma of all subjects was collected within 72 hours after birth.The expression levels of cytokines IL-2,IL-4,IL-6,IL-9,IL-10,IL-21,IFN-γ and TNF-α were determined,and cytokines with high expression levels(high correlation)were screened out.Receiver operating characteristic(ROC)curve was used to analyze the specificity and sensitivity of the selected cytokines in the diagnosis of neonatal early-onset sepsis.Results Among the 8 cytokines mentioned above,the concentrations of IL-6,IL-9 and IL-21 in cord blood plasma of neonatal early-onset sepsis positive blood culture patients(392.6±258.7pg/ml,11.9±7.5pg/ml,29.1±16.8 pg/ml)and negative blood culture patients(353.8±244.5pg/ml,10.4±6.3pg/ml,27.7±19.2pg/ml)were higher than those of the control group(34.9±25.1pg/ml,5.9±4.5pg/ml,10.8±10.1 pg/ml),with significant differences(t=23.961,20.732;15.174,17.824;22.466,21.193,all P<0.01),and the increase of IL-6 concentration was the most obvious.ROC curve analysis(the cut-off values of IL-6,IL-9 and IL-21:123.0 pg/ml,3.60 pg/ml,6.00 pg/ml,respectively)showed that the areas under the ROC curve for IL-6,IL-9 and IL-21 alone detection were 0.876(95%CI:0.786~0.955),0.782(95%CI:0.667~0.875)and 0.825(95%CI:0.737~0.913),respectively.The area under the ROC curve for the combined detection of IL-6,IL-9 and IL-21 was 0.930(95%CI:0.875~0.997).The combined detection of IL-6,IL-9 and IL-21 improved the specificity and sensitivity of the test than IL-6,IL-9 and IL-21 alone detection,and the differences were statistically significant(Z=2.137,2.391,2.257,all P<0.05).There was no significant difference in cytokine expression between positive blood culture and negative blood culture neonates with early-onset sepsis(t=0.276~3.377,all P>0.05).Conclusion The cytokines expression of IL-6,IL-9 and IL-21 in neonatal umbilical cord plasma of neonatal early-onset sepsis were increased.Combined detection of IL-6,IL-9 and IL-21 has good diagnostic value for early-onset neonatal sepsis.
4.Influence of Xin'an River water conservancy project on epidemic of schistosomiasis in the Qiandao Lake reservoir area
Weimin XU ; Yihong ZHENG ; Zhaoying QIAN ; Sujuan ZHU ; Xiaolan YAN ; Mingjiang WU ; Yibiao ZHOU ; Baofeng QIAN ; Hua DING ; Jianfeng ZHANG ; Qingwu JIANG ; Liyong WEN
Chinese Journal of Endemiology 2018;37(5):414-419
Objective To study epidemic of schistosomiasis in the Qiandao Lake reservoir area,and to provide a reference for prevention and control of schistosomiasis in the construction of large water conservancy projects in the epidemic area of schistosomiasis.Methods The data over the years of snail condition and monitoring of schistosomiasis before and after building the dam,and water conservancy project reconstruction related information were collected.Based on the survey results of the river channel,the lake beach and the dissipation zone in the reservoir area,the influence of Xin'an River water conservancy project on epidemic of schistosomiasis in the Qiandao Lake reservoir area was analyzed,and the epidemic factors of the schistosomiasis in the Three Gorges reservoir were compared and analyzed.Results Before the dam was built,an area of 38 144 000 m2 was examined but Oncomelania was undetected.The Qiandao Lake reservoir area belonged to a non epidemic area of schistosomiasis.After the dam was built,557 cases of schistosomiasis were found in 6 232 immigrants during 1962-1965,resulting in an imported epidemic.In 1970-1980,an area of 379 654 m2 in which Oncomelania was found was examined and snails were mainly distributed in some rice fields and ditches in the end of the reservoir.949 cases of local schistosomiasis were found in the snails.The condition and condition of the snail are gradually controlled through several decades of comprehensive prevention and control.Compared with the epidemic factors of schistosomiasis in Qiandao Lake and the Three Gorges reservoir,the environment of elevation beach and ecologically fragile fluctuation zone coexist in the two reservoir areas.Conclusion From the long-term longitudinal monitoring data of the Qiandao Lake reservoir area and the epidemic regularity of schistosomiasis and the comparison with the ecology of the Three Gorges reservoir,it is concluded that the two reservoir areas will not cause a large range of schistosomiasis epidemic in general,but it does not exclude the possibility of the breeding of the inputting Oncomelania.
5.Pharmacometabolomics and mass spectrometry imaging approach to reveal the neurochemical mechanisms of Polygala tenuifolia
Li QIAN ; Bai JINPENG ; Ma YUXUE ; Sun YU ; Zhou WENBIN ; Wang ZHAOYING ; Zhou ZHI ; Wang ZHONGHUA ; Chen YANHUA ; Abliz ZEPER
Journal of Pharmaceutical Analysis 2024;14(7):1035-1046
Polygala tenuifolia,commonly known as Yuanzhi(YZ)in Chinese,has been shown to possess anti-insomnia properties.However,the material basis and the mechanism underlying its sedative-hypnotic effects remain unclear.Herein,we investigated the active components and neurochemical mechanism of YZ extracts using liquid chromatography tandem mass spectrometry(LC-MS/MS)-based pharmaco-metabolomics and mass spectrometry imaging(MSI)-based spatial resolved metabolomics.According to the results,17 prototypes out of 101 ingredients in the YZ extract were detected in both the plasma and brain,which might be the major components contributing to the sedative-hypnotic effects.Network pharmacology analysis revealed that these prototypes may exert their effects through neuroactive ligand-receptor interaction,serotonergic synapse,dopaminergic synapse,and dopaminergic synapse,among other pathways.LC-MS/MS-based targeted metabolomics and Western blot(WB)revealed that tryptophan-serotonin-melatonin(Trp-5-HT-Mel)and tyrosine-norepinephrine-adrenaline(Tyr-Ne-Ad)are the key regulated pathways.Dopa decarboxylase(DDC)upregulation and phenylethanolamine N-methyltransferase(PNMT)downregulation further confirmed these pathways.Furthermore,MSI-based spatially resolved metabolomics revealed notable alterations in 5-HT in the pineal gland(PG),and Ad in the brainstem,including the middle brain(MB),pons(PN),and hypothalamus(HY).In summary,this study illustrates the efficacy of an integrated multidimensional metabolomics approach in unraveling the sedative-hypnotic effects and neurochemical mechanisms of a Chinese herbal medicine,YZ.