Distribution and exposure assessment of phthalic acid esters （PAEs） in indoor dust of Shanghai / 上海预防医学

Objective To characterize the distribution and assess the exposure to phthalic acid esters （PAEs） in the indoor dust of Shanghai City. Methods Samples were collected from 33 sampling sites， including homes， hotels， offices and public places， in Shanghai in 2018， 2019， and 2020. The samples were pretreated by 100 sieves， extracted and concentrated， and then analyzed by gas chromatography-mass spectrometry in selected ion mode （SIM）. Results Results on the characteristics of PAEs in indoor dust in different places showed that concentrations of PAEs were in a range of <0.01-2 464 mg·kg-1.The average concentration of 16 PAEs was 613 mg·kg-1. Bis（2-ethylhexyl） phthalate （DEHP）， di-iso-butyl phthalate （DiBP）， di-n-butyl phthalate （DBP） and di-n-octyl phthalate （DnOP） were the main components of PAEs in indoor dust， accounting for approximately 99.5% of 16 PAEs. The intake of DEHP， DBP， DEP and BBP was lower than the tolerable daily intake （TDI） and reference doses （RfD） set by EU CSTEE and U.S. EPA. Conclusion Average daily dose （ADD） via indoor dust is estimated， and the order of intake through different pathways is hand-oral intake>skin contact>respiratory inhalation. Exposure risk of PAEs in children is greater than that in adults.

Distribution and exposure assessment of phthalic acid esters （PAEs） in indoor dust of Shanghai / 上海预防医学

Objective To characterize the distribution and assess the exposure to phthalic acid esters （PAEs） in the indoor dust of Shanghai City. Methods Samples were collected from 33 sampling sites， including homes， hotels， offices and public places， in Shanghai in 2018， 2019， and 2020. The samples were pretreated by 100 sieves， extracted and concentrated， and then analyzed by gas chromatography-mass spectrometry in selected ion mode （SIM）. Results Results on the characteristics of PAEs in indoor dust in different places showed that concentrations of PAEs were in a range of <0.01-2 464 mg·kg-1.The average concentration of 16 PAEs was 613 mg·kg-1. Bis（2-ethylhexyl） phthalate （DEHP）， di-iso-butyl phthalate （DiBP）， di-n-butyl phthalate （DBP） and di-n-octyl phthalate （DnOP） were the main components of PAEs in indoor dust， accounting for approximately 99.5% of 16 PAEs. The intake of DEHP， DBP， DEP and BBP was lower than the tolerable daily intake （TDI） and reference doses （RfD） set by EU CSTEE and U.S. EPA. Conclusion Average daily dose （ADD） via indoor dust is estimated， and the order of intake through different pathways is hand-oral intake>skin contact>respiratory inhalation. Exposure risk of PAEs in children is greater than that in adults.

Inhibition of choriocarcinoma by Fe3O4-dextran-anti-ß-human chorionic gonadotropin nanoparticles containing antisense oligodeoxynucleotide of heparanase.

OBJECTIVE: To observe the influence of Fe3O4-dextran-anti-ß-human chorionic gonadotropin (HCG) carrying heparanase (Hpa) antisense oligodeoxynucleotide (ASODN), via the invasion, proliferation, and Hpa expression of JEG-3 cell lines and inhibitory effect of transplanted choriocarcinoma tumor growth. METHODS: The different abilities of invasion and proliferation between transfected JEG-3 and untransfected JEG-3 were measured by Matrigel invasion assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in vitro. The effect of Hpa ASODN transfection on the expression of Hpa mRNA and protein was measured by reverse-transcription polymerase chain reaction and Western blot. The transplanted choriocarcinoma tumors were taken out to calculate the inhibitory effect on tumor growth of Hpa ASODN. RESULTS: IN THIS STUDY, WE FOUND THAT: (1) the invasive ability of JEG-3 cells was inhibited sufficiently (P < 0.05) after JEG-3 cells were transfected by Fe3O4-dextran-anti-ßHCG carrying Hpa ASODN; (2) after JEG-3 cells were transfected by Fe3O4-dextran-anti-ßHCG carrying Hpa ASODN at 48 and 72 hours, the proliferative ability of JEG-3 cells was inhibited sufficiently (P < 0.05); (3) the expression of Hpa mRNA and protein in JEG-3 cells was inhibited efficiently after JEG-3 cells were transfected by Fe3O4-dextran-anti-ßHCG carrying Hpa ASODN (P < 0.05); and (4) Fe3O4-dextran-anti-ßHCG carrying Hpa ASODN had an inhibitory effect on the transplanted choriocarcinoma tumor growth (P < 0.05) and was harmless on nude mice. CONCLUSION: Fe3O4-dextran-anti-ßHCG carrying Hpa ASODN weakened the invasive and proliferative ability of choriocarcinoma, with a significant inhibitory effect on the transplanted choriocarcinoma tumor. Therefore, Fe3O4-dextran-anti-ßHCG carrying Hpa ASODN is an effective gene therapy, and Fe3O4-dextran-anti-ßHCG nanoparticles are a harmless and effective gene vector.

Study of the relationship between Wnt/β-catenin and drug resistance of choriocarcinoma / 中国医师杂志

Objective To investigate the relationship between the expression of β-catenin and drug-resistance mechanism of choriocarcinoma according to the expression of β-catenin in JEG-3 cells (human choriocarcinoma cell line) and drug resistant JEG-3/VP16 cells.Methods The mRNA and protein expressions of β-catenin were analyzed with reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting.Flow cytometry was used to determine the percentages of β-catenin-positive cells in the two choriocarcinoma cell lines.Results Both drug resistant choriocarcinoma cells and drag sensitive cells were found to express β-catenin; but the expression of β-catenin mRNA (1.43 ±0.24) and protein(1.49 ±0.17)in drug resistant choriocarcinoma cells was found much higher than that in drug sensitive cells(0.65 ±0.14,0.66 ±0.16,P ＜0.01).And according to detect by flow cytometry,we found the number of β-catenin-positive cells in JEG-3/VP16 cells [(40.13 ±5.17) ％] was much more than that in JEG-3 cells [(13.15 ± 1.48) ％,P ＜ 0.01].Conclusions β-catenin was highly expressed in the drug resistant choriocarcinoma cell line (JEG-3/VP16).It indicates β-catenin might be involved in the drug resistance mechanism of choriocareinoma.

Preparation and characterization of magnetic nanoparticles containing Fe(3)O(4)-dextran-anti-ß-human chorionic gonadotropin, a new generation choriocarcinoma-specific gene vector.

OBJECTIVE: To evaluate the feasibility of using magnetic iron oxide (Fe(3)O(4))-dextran-anti-ß-human chorionic gonadotropin (HCG) nanoparticles as a gene vector for cellular transfections. STUDY DESIGN: Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles were synthesized by chemical coprecipitation. The configuration, diameter, and iron content of the nanoparticles were detected by transmission electron microscopy (TEM), light scatter, and atomic absorption spectrophotometry. A3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide assay was used to evaluate the cytotoxicity of Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles. Enzyme-linked immunosorbent assay and indirect immunofluorescence were used to evaluate immunoreactivity. The efficiency of absorbing DNA and resisting deoxyribonuclease I (DNase I) digestion when bound to Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles was examined by agarose gel electrophoresis. The ability of Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles to absorb heparanase antisense oligodeoxynucleotides (AS-ODN) nanoparticles in different cell lines was evaluated by flow cytometry. The tissue distribution of heparanase AS-ODN magnetic nanoparticles in choriocarcinoma tumors transplanted in nude mice was detected by atomic absorption spectrophotometry. RESULTS: TEM demonstrated that the shape of nanoparticles is irregular. Light scatter revealed nanoparticles with a mean diameter of 75.5 nm and an iron content of 37.5 µg/mL. No cytotoxicity was observed when the concentration of Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles was <37.5 µg/mL. Fe(3)O(4)-dextran nanoparticles have a satisfactory potential to combine with ß-HCG antibody. Agarose gel electrophoresis analysis of binding experiments showed that after treatment with sodium periodate, Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles have a satisfactory potential to absorb DNA, and the protection experiment showed that nanoparticles can effectively protect DNA from DNase I digestion. Aldehyde Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles can transfect reporter genes, and the transfection efficiency of these nanoparticles is greater than that of liposomes (P < 0.05). Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles can concentrate in choriocarcinoma cells and in transplanted choriocarcinoma tumors. CONCLUSIONS: The results confirm that Fe(3)O(4)-dextran-anti-ß-HCG nanoparticles have potential as a secure, effective, and choriocarcinoma-specific targeting gene vector.

Proteomics reveals protein profile changes in cyclooxygenase-2 inhibitor-treated endometrial cancer cells.

OBJECTIVE: To examine effects of an inhibitor of cyclooxygenase (COX)-2, NS-398, on the proliferation, apoptosis and invasion characteristics of endometrial cancer cell RL95-2. METHODS: (1) Western blotting was carried out to determine COX-2 protein expression in RL95-2 cells and normal endometrium specimens. (2) The effect of NS-398 treatment on the cell proliferation, apoptosis, and invasion was assessed by methyl thiazolyl tetrazolium assay, flow cytometry, and matrigel invasion assay, respectively. (3) Finally, the proteomic analysis was used to find out proteins that are differentially expressed because of NS-398 treatment. RESULTS: (1) COX-2 protein in RL95-2 cell line was significantly higher than that in normal endometrium. (2) NS-398 had significant growth inhibition effects on RL95-2 cells in a dose- and time-dependent manner. (3) NS-398 increased the proportion of cells in G1 and decreased the proportion of cells in the G2 phase in RL95-2 cells. (4) NS-398 could restrain endometrial cancer cells invasion. (5) The proteomic analysis revealed several proteins that are differentially expressed because of NS-398 treatment; the down-regulated proteins identified are hnRNP K, alpha enolase, Hsp70, tropomyosin, and protein disulfide isomerase, the up-regulated protein is phosphatidylethanolamine binding protein. CONCLUSIONS: The expression of COX-2 plays an important role in tumorigenesis of endometrial cancer. NS-398 can inhibit the ability of RL95-2 cell proliferation, viability, and invasion. In this study, the well-resolved reproducible 2-DE maps of NS-398 treated and control RL95-2 cells were established, and the significantly different expressed proteins are preliminary identified.

Expression of heparanase and angiopoietin-2 in patients with endometriosis.

OBJECTIVE: The objective was to investigate the expression of heparanase (Hpa) and angiopoietin-2 (Ang-2) in endometriosis. STUDY DESIGN: In ectopic and eutopic endometrium of patients undergoing laparoscopy for endometriosis (n=86) and in normal endometrium of patients undergoing laparoscopic tubal ligation or hysteroscopic resection because of uterus septus (n=30), we determined Hpa and Ang-2 gene expression by RT-PCR. To support the mRNA data, the expression of Hpa and Ang-2 protein was measured by Western blot analysis. Finally, Hpa and Ang-2 in these tissues was localized by immunohistochemical staining. RESULT(S): The positive rate of Hpa and Ang-2 mRNA in ectopic and eutopic endometrium in the study group was significantly higher than that in normal endometrium in the control group. In the study group, ectopic and eutopic endometrium expressed a higher positive rate of Hpa and Ang-2 protein, whereas in the control group, normal endometrium expressed a lower positive rate of Hpa and Ang-2 protein. In eutopic and ectopic endometrium, there was balanced expression between Hpa and Ang-2. Both Hpa and Ang-2 showed a balanced expression between eutopic and ectopic endometrium. In ectopic endometrium, strong staining for Hpa and Ang-2 was observed both in epithelial cells and in stromal cells, but in eutopic endometrium, Hpa and Ang-2 were mainly expressed in epithelial cells. CONCLUSION: The higher expression of Hpa and Ang-2 in ectopic and eutopic endometrium may play an important role in the pathogenesis and development of endometriosis.

Heparanase expression correlates with metastatic capability in human choriocarcinoma.

OBJECTIVE: In this report, we studied the role of Hpa in metastatic capability of human choriocarcinoma. At the same time, we investigated the effect of Hpa antisense oligodeoxynucleotide (ASODN) on inhibition of invasiveness of human choriocarcinoma. METHODS: The different invasion ability between JEG-3 and JAR cell lines was proved by Matrigel invasion assay in vitro. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analyses were carried out respectively to determine Hpa gene and protein expression; the localization of this molecule was demonstrated by immunohistochemistry. Finally, Hpa antisense oligodeoxynucleotide (ASODN) was transfected into JEG-3 cells and Hpa mRNA and protein were quantified by RT-PCR and Western blot. The effect of ASODN on the metastatic capability of JEG-3 was evaluated by Matrigel invasion assay. RESULTS: (1) We proved that the invasion ability of JEG-3 cell line was stronger than that of JAR cell line (P<0.05). (2) We found that the Hpa gene and protein in JEG-3 and JAR cell lines were significantly higher than those in normal chorion (P<0.05). On the other hand, we detected that JEG-3 expressed much more Hpa than JAR (P<0.05). (3) Both in JEG-3 cell and in JAR cell, we found that Hpa protein express in cytoplasm. (4) After transfection of Hpa ASODN, Hpa mRNA and protein expression in JEG-3 cell decreased 4- and 5-fold. At the same time, we also observed that the invasion ability of JEG-3 cell was weakened than before (P<0.05). CONCLUSION: The current study demonstrated that the expression of Hpa plays an important role in metastatic capability of human choriocarcinoma and reducing the expression of Hpa can help weaken the invasion ability of human choriocarcinoma.