Optimization of chromatographic conditions for the determination of clenbuterol hydrochloride residues in animal edible product / 预防医学

Objective To establish a high performance liquid chromatography (HPLC) through the optimization of the chromatographic conditions, which can detect the contents of clenbuterol hydrochloride (CL) residues in animal edible product in a large quantity. Methods The animal edible product were extracted by perchloric acid, and then impurities were removed by liquid-liquid extraction (LLE) which used ethyl acetate- isopropanol. After the organic phase was concentrated, C18 column (150 mm×4.6 mm, 5 μm) was used to separate CL. Mobile phase were methanol-sodium dihydrogen phosphate, and then determined by HPLC. Results A good linear response was obtained over the range of 0.2-10.0 μg/mL with the correlation coefficient (r) 0.99984. The method determination limit was 0.15 μg/kg which was lower than the National standard method 0.5μg/kg. The retention time of the CL was 6.51 min, the chromatographic peak was good. The recovery rates spiked with standards 1.6-12 μg were 92.86％-100.93％, which was higher than National standard method (89.79％-92.36％) . The precision of intra-day and inter-day were both under 5％, which lower than National standard. Conclusion The optimized chromatographic conditions are suitable for the large quantity determination of clenbuterol hydrochloride in animal edible product.

Expression of HK2 in prostate cancer and its effect on malignant phenotype of prostate cancer cells / 临床与实验病理学杂志

Purpose To study the expression of HK2 in human prostate cancer (PCa) tissues and its effect on malignant phenotype of prostate cancer cells.Methods HK2 expression in PCa tissues was determined by microarray database and immunohistochemical staining.Subsequently,the change of cellular phenotype was detected by glycometabolism kit,CCK-8 kit,and flow cytometry after HK2 knockdown.Results HK2 expression was elevated followed by prostate cancer development.HK2 depletion inhibited cellular proliferation and aerobic glycolysis,and increased the ratio of early apoptosis.Conclusion HK2 expression increases in the process of PCa malignant progression.It plays a critical role in cellular proliferation,glycometabolism,and apoptosis,the mechanism of which needs further exploration.

An investigation on total radioactivity in drinking water around operating Qinshan nuclear power plant / 预防医学

Objective To investigate the effect on radioactivity in drinking water around Qinshan nuclear power station (QNPS)in normal operational condition.Methods The field monitoring and laboratory analysis methods were adopted to detect the total radioactivity level in drinking water in 2015,according to different distances from the nuclear island and different types of water.Results The total alpha and total beta radioactivity level in drinking water around QNPS were 0.027(0.098)Bq /L and 0.263(0.071)Bq /L respectively,which were obviously lower than the national health standard limits(total alpha and total beta are 0.5,1.0 Bq /L respectively).Total radioactivity level had no relation with the distance from the nuclear island (P >0.05).The total alpha radioactivity in deep well water was the highest among the investigated three types of drinking water,and the highest value was 0.224 Bq /L.The beta radioactivity level in river water was the highest,and the highest value was 0.408 Bq /L.The total alpha radioactivity level was 0.017 (0.013)Bq /L in 2015, higher than the average level during 2010—2014.The beta radioactivity average level was 0.319 (0.102)and 0.289 (0.055)Bq /L,also higher than the average level during 2010—2014.Conclusion The total radioactivity in drinking water among nuclear power plant is in normal background level,so at present there is no effect of the radioactive contamination on drinking water around QNPS in nuclear power plant's normal operational condition.

A selectivity study of the examination of radionuclides Cesium-1 37 for foods based on different detection conditions / 预防医学

Objective Aselectivitystudywasconductedthroughtheexaminationofradionuclides137Cs(Cesium-137)for foodbasedondifferentdetectionconditions.Methods Atotalof48foodsampleswereselectedfromthreeareasincluding Qinshan nuclear power plant,Sanmen nuclear power plant and Hangzhou and Zhoushan respectively.1 37 Cs of these samples were determined by γspectrometry and Phosphoric acid ammonium molybdate method.The level of 48 foods were statistically analyzed,and then the time consuming,sample size requirements,influence factors were comprehensively discussed,thustheselectionreferenceproposaloftheexaminationmethodcouldbeprovided.Results Therewereno significant difference for the data of two examination method (P>0.05 ).The limit of detection of the γspectrometry was lower (P <0.05 ).Compared with Phosphoric acid ammonium molybdate method,γspectrometry had lower limit of detection,and could detect a variety of radionuclides at a time,but need more sample and time-consuming when multi-sample were detected.The limit of detection of the Phosphoric acid ammonium molybdate method was high,and the chemicalprocessingstepswerecumbersomeandwaseasytobeinterferedby134Cs.Conclusion Thelimitofdetectionof the γspectrometry is low,and the sensitivity of the Phosphoric acid ammonium molybdate method is high.Most food are recommended to be detected by γspectrometry in the practical work,and the food which were difficult collected,less ash or low content,are recommended to be detected by Phosphoric acid ammonium molybdate method.

Clinical analysis of extraperitoneal laparoscopic radical prostatectomy / 中国综合临床

Objective To evaluate the feasibility and the clinical value of extraperitoneal laparoscopic radical prostatectomy in treatment of localized prostate cancer.Methods Clinical data of 26 patients with localized prostate cancer treated with extraperitoneal laparoscopic radical prostatectomy were analyzed retrospectively.All patients were pathologic diagnosed with prostate cancer by preoperative prostate biopsy or transurethral resection of prostate surgery.Gleason grade was from 6-8.Results Twenty-six operations were successfully accomplished ,without converting to open approach.The operative time was 120-270 min(mean was 165 min) ,the intraoperative blood loss was 180-650 ml (mean was 320 ml) ,indwelling catheter time 12-19 d (mean was 14 d).There were 6 cases with little uroclepsia, satisfactory with urination after contract urethral sphincter for 1-3 Months.Pathologically confirmed all prostate cancer;2 cases of positive margins after surgery plus endocrine therapy.All the cases were followed up from 2 to 36 months.The biochemical recurrence was 5 cases who had undergone endocrine therapy.Conclusion Extraperitoneal laparoscopic radical prostatectomy is a safe and feasible procedure with little trauma, small bleeding and fast recovery which is well worth popularizing.Replace open surgery may become frist choice therapeutic method for localized prostate cancer.

Expressions and clinical significance of chemokine receptor-4 and vascular endothelial growth factor in renal cell carcinoma / 中国综合临床

Objective To study the role and clinical significance of chemokine receptor-4 (CXCR4) and vascular endothelial growth factor (VEGF) in the occurrence and development of renal cell carcinoma. Methods Expression of CXCR4 and VEGF were detected by SP immunohistochemical technique in 56 cases of kidney carcinoma tissues (including 20 cases of lymph node metastasis), 10 normal tissues nearby kidney cancer. Results The positive rates of CXCR4 and VEGF were 66. 1% (37/56) and 73. 2% (41/56),which were significantly higher than those in normal tissues( 20. 0% (2/10) and 30. 0% (3/10), respectively) (P ＜ 0. 05 ＝. The expression of CXCR4 protein was significantly positively correlated with that of VEGF protein (r = 0. 315 ,P ＜ 0.05 ＝ in renal cell carcinoma. The expression of CXCR4 and VEGF was closely related to stages of tumor ( χ2 = 9. 520, P = 0. 023; χ2 = 9. 072, P = 0. 027 ), lymphatic metastasis, degree of invasion ( χ2 =4. 972, P = 0. 026; χ2 = 3.910, P = 0. 034 ), and microvessel density ( MVD) ( P ＜ 0. 05 ＝. However, they were not related to sex ( χ2 = 0. 020, P= 0. 887; χ2 = 0. 001, P = 0. 716 ), tumor size ( χ2 = 0. 003, P = 0. 995; χ2 =0. 108, P = 0. 990) and pathologic types ( χ2 = 1. 960, P = 0. 900; χ2 = 0. 112, P = 0. 994). Conclusion There is a significant positive correlation between high expressions of CXCR4 and VEGF proteins in renal cell carcinoma,the high expressions of CXCR4 and VEGF proteins may be related to the metastasis and prognosis of renal cell carcinoma,thus they could be used as important indicators in judging the metastasis prognosis of renal cell carcinoma,and offer prospects for the treatment of renal cell carcinona.

Stromal cells inhibit Kallikrein 7 expression in the epithelial cells of benign prostate hyperplasia / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the influence of stromal cells on the Kallikrein 7 (KLK7) expression of epithelial cells in benign prostate hyperplasia (BPH).</p><p><b>METHODS</b>We constructed a stromal-epithelial co-culture model after separating the two types of cells from BPH tissues and identifying them by cell morphology and chemiluminescent microparticle immunoassay (CMIA). The expression of KLK7 mRNA was detected by RT-PCR in the epithelial cells with or without the stromal cells, and that of the KLK7 protein (hK7) determined by Western blot.</p><p><b>RESULTS</b>Stromal and epithelial cells were successfully separated and identified, and a stromal-epithelial co-culture model successfully established. RT-PCR showed that the mRNA expression of the KLK7 gene was higher in the epithelial cells co-cultured with stromal cells than in the epithelial cells alone, and the gray value of KLK7 to GAPDH was 1.41 +/- 0.041 in the former and 1.78 +/- 0.10 in the latter (P < 0.01). The results of Western blot were consistent with those of RT-PCR.</p><p><b>CONCLUSION</b>Stromal cells can suppress the expression of the KLK7 gene in the epithelial cells in BPH. KLK7 may be involved in the change of epithelial cells stimulated by stromal cells.</p>

Detecting DNA damage of human lymphocytes exposed to 1,2-DCE with γH2AX identified antibody using flow cytometer assay / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study DNA damage of human peripheral blood lymphocytes exposed to 1,2-dichloroethane (1,2-DCE) with flow cytometry (FCM) assay.</p><p><b>METHODS</b>The lymphocytes were obtained from 21 workers who are occupationally exposed to 1,2-DCE (exposed group) and 27 workers who were not exposed to 1,2-DCE in the same factory (inner control) and 28 island residents who had never been occupationally exposed to adverse factors (external control). FCM assay was adopted to detect DNA damage of the lymphocytes of each group. Lymphocytes of the health people were incubated with 1,2-DCE at different doses, and FCM assay was used to detect DNA damage.</p><p><b>RESULTS</b>DNA damage rate (%) of the exposed group of exposed workers (4.05% ± 2.55%) was significantly higher than the inner control group of workers (1.97% ± 1.40%) and external control groups of island residents (0.23% ± 0.13%), and the DNA damage of inner control was higher than the external control, all the differences were statistically significant (P < 0.01 or P < 0.05). The geometric mean fluorescence intensity of the workers in the exposed group (3.33 ± 3.01) was significantly higher than the (2.07 ± 0.58) only (P < 0.05). There was no significant difference in the DNA damage rate as well as the geometric mean fluorescence intensity among the exposed group of workers with different years of working period (P > 0.05). In vitro, the fluorescence intensity at the dose of 20, 30 µmol/L for 0.5 h exposure showed statistical significance compared with the negative control group (P < 0.01). The DNA damage rate at the dose of 20, 30 µmol/L for 1.0 h exposure was statistically significant compared with the negative control group (P < 0.05, P < 0.01); The fluorescence intensity at the dose of 10, 20, 30 µmol/L for 1.0 h exposure was statistically significant compared with the negative control group (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>1,2-DEC can cause DNA damage. And γH2AX FCM assay can be a sensitive, objective and effective method of detecting DNA damage of peripheral blood lymphocytes.</p>

The curative effects of different drugs on liver cell damage of rats induced by acute nickel carbonyl poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To assess the curative effects of different drugs on liver cell damage of rats induced by acute nickel carbonyl poisoning.</p><p><b>METHODS</b>In present study 220 SD rats were divided into control group (10 rats), carbonyl nickel group (10 rats), 20 mg/kg methylprednisolone group (40 rats), 100 mg/kg DDC group (40 rats), 10 µmol/kg sodium selenite group (40 rats), 0.25 ml shenfuhuiyangtang group (40 rats) and 20 mg/kg methylprednisolone with 100 mg/kg DDC group (40 rats). All rats except for control group inhaled passively 250 mg/m(3) carbonyl nickel for 30 minutes. At 4h and 30h after exposure, the drugs were given intraperitoneally to the rats. On the 3rd and 7th days after exposure, the liver samples were taken from 10 rats each group. The DNA damage of liver cells was detected using comet assay, the ultrastructure changes in liver cells were examined under an electronmicroscope.</p><p><b>RESULTS</b>Compared to carbonyl nickel group, the tail lengths of liver cells in 5 groups administrated at 4 h or 30 h and tested on the 3rd or 7th day after exposure decreased significantly (P < 0.05). Compared to the control group, the tail lengths of liver cells in sodium selenite and shenfuhuiyangtang groups administrated at 4h after exposure or sodium selenite, shenfuhuiyangtang and methylprednisolone with DDC groups administrated at 30h after exposure increased significantly (P < 0.05 or P < 0.01), when tested on the 3rd day after exposure. Except from methylprednisolone sub-group administrated at 4h and tested on the 7th day after exposure, the tail lengths of liver cells in other groups administrated at 4 h or 30 h and tested on the 7th day after exposure increased significantly (P < 0.05). Compared to carbonyl nickel group, the Olive moment of liver cells in 5 groups administrated at 4 h or 30 h tested on the 3rd or 7th day after exposure decreased significantly (P < 0.05 or P < 0.01). Compared to the control group, the Olive moment of liver cells in following groups (selenite and shenfuhuiyangtang groups administrated at 4 h or 30 h and tested on the 3rd or 7th day after exposure, DDC group administrated at 4 h or 30 h and tested on the 7th day after exposure, DDC group administrated at 30h and tested on the 3rd day after exposure, and methylprednisolone with DDC group administrated at 30 h and tested on the 7th day after exposure) increased significantly (P < 0.05 or P < 0.01). As compared with carbonyl nickel group, the ultrastructure observation indicated that the nucleus and other organelles of liver cells in methylprednisolone, DDC and methylprednisolone with DDC groups administrated at 4h and tested on the 3rd day were access to normal levels.</p><p><b>CONCLUSION</b>The results of present study showed that methylprednisolone, DDC and methylprednisolone with DDC could improve obviously the repair of rat liver cell damage induced by acute carbonyl nickel poisoning, and the curative effects of early treatment were better than those of later treatment.</p>

Detection of DNA damage of workers occupationally exposed to lead with flow cytometer assay / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study DNA damage of workers occupationally exposed to lead with flow cytometer assay.</p><p><b>METHODS</b>The lymphocytes were obtained from 41 workers occupationally exposed to lead (comparable group) and another 50 from control group. Flow cytometer (FCM) assay was used to detect DNA damage.</p><p><b>RESULTS</b>DNA damage rate and geometric mean fluorescence intensity in the comparable group were significantly higher than those in the control group (P<0.05). There were no significant differences in the DNA damage and geometric mean fluorescence intensity between different age groups (P>0.05). The differences in correlation analysis between blood lead, urine lead, delta-ALA and DNA damage rate were not significant (P>0.05). The correlation analysis showed no statistical significance between concentration of blood lead, urine lead, delta-ALA and geometric mean fluorescence intensity (P>0.05). There was positive correlations not only between the high concentration of blood lead, delta-ALA and damage rate of DNA, but also between the high concentration of blood lead and geometric mean fluorescence intensity. The coefficient r showed statistical significance (P<0.05).</p><p><b>CONCLUSION</b>Occupational lead exposure can cause DNA damage. Gamma H2AX flow cytometer assay is a sensitive, objective and effective method for detection of DNA damage of peripheral blood lymphocytes.</p>

Down-regulation of kallikrein in prostate cancer / 中华泌尿外科杂志

Objective To study the expression of kallikrein 7 (KLK7) in different prostate tissues and its clinical significance. Methods KLK7 mRNA levels in normal prostate epithelia (5 cases), benign prostat(ic) hyperplasia (BPH) epithelia (13 cases), prostate cancer and prostate cancer cell lines (8 cases) were analyzed by using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Western blot was used to analyze the protein levels of human kallikrein 7 (hK7) in benign prostate epithelia and prostate cancer cell lines, hK7 expressions were examined in 20 normal prostate tissue specimens, 50 BPH specimens and 103 prostate cancer specimens by immunohistochemical staining.Results The mRNA levels of KLK7 in normal prostate, BPH and prostate cancer were 0.59, 0.52 and 0.02 respectively, mRNA levels of KLK7 were significantly different among the three groups (F=13.03, P＜0.01). mRNA levels of KLK7 were decreased in prostate cancers compared with that in benign hyperplastic prostate epithelial cells (P＜0.01) and in normal prostate epithelial cells (P＜0.01). No significant difference of KLK7 mRNA levels was found between normal prostate and BPH. The protein levels of KLK7 in normal prostate, BPH, DU145, LNCaP, PC3,22RV1 and BPH1 was 0.22, O. 40, 0.01, 0.05, 0, 0.03 and 0.14 respectively, hK7 protein level was down-regulated in prostate cancer cell lines compared to benign prostate epithelial cells. The expression of bK7 was observed in benign prostate epithelial cells, whereas little or no staining was observed in prostate cancer cells in immunohistochemical study, hK7 protein was detected in 13 of 20 (65%)normal prostate specimens, 38 of 50 (76%) BPH specimens and 18 of 103 (17.5%) prostate cancer specimens. The difference between the normal prostate and prostate cancer was significant (Z=-4.43, P＜0.01). The difference between BPH and prostate cancer was significant (Z=-7.77,P＜0.01) as well. However, no significant difference of hK7 protein level was found between normal prostate and BPH (Z=-1. 52, P＞0.05). Conclusions KLK7 expression level is down regulated in prostate cancer. KLK7 may play an important role in prostate cancer progression.

DMF-induced adult hepatocyte apoptosis and its effects on expression of Bcl-2, Bax and Caspase-3 / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the apoptosis-inducing effect of DMF on the human liver cells (HL-7702) in vitro.</p><p><b>METHODS</b>Liver cells were exposed to different concentrations of DMF (0, 50, 100, 150, 200 mmol/L) for 12 hours. Apoptotic rate, the expression of Bax, Bcl-2 and Caspase-3 in liver cells were measured by FCM and western blotting respectively.</p><p><b>RESULTS</b>The increase in apoptotic rate of hepatocytes in concentration-manner was shown after DMF treatment for 12 h. After treatment the expression of Bcl-2 was decreased steadily and lower than the control group (P < 0.01), the expression of Bax showed no significant difference among the groups of different dosage by one-factor analysis of variance (P > 0.05), as the increase of the dosage of DMF. The ratio of Bcl-2/Bax dropped with the dosage of DMF increasing, and the ratio in 200 mmol/L of DMF was significantly lower than that of the control (P < 0.01). The new lands of procaspase-3 in 150, 200 mmol/L were observed, which demonstrated that there was active caspase-3.</p><p><b>CONCLUSION</b>DMF can induce apoptosis of cultured adult normal hepatocytes in vitro, and the mechanism might be related to the decrease of Bcl-2/Bax and the cleavage of Caspase-3.</p>