[Determination of platinum anticancer drugs in occupational settings by ICP-MS].

Objective: To establish an inductively coupled plasma mass spectrometry (ICP-MS) for platinum antineoplastic drugs in the environment. Methods: The platinum antineoplastic drugs in the environmental table were eluted by wiping and collecting pure water, and the supernatant was taken by centrifugation and inductively coupled plasma mass spectrometry for detection. Results: The concentration range of 0-8.0 µg/L was good, the correlation coefficient was 1.000, the detection limit was 0.0006 µg/L, the lower quantitative limit was 0.002 µg/L, the method precision was between 0.9%-1.3%, and the sample standard recovery rate was between 97.0%-98.5%. Conclusion: This method has low detection limit, high accuracy and precision, and simple sample pretreatment, which is suitable for the determination of platinum antineoplastic drugs in environmental tables.

[Liquid chromatography-tandem mass spectrometry method for the determination of paraquat and diquat in plasma and urine].

Objective: To establish a LC-MS/MS method for determination of paraquat and diquat in plasma and urine samples. Methods: Plasma is precipitated by acetonitrile then diluent with phosphate buffer (pH=7) , urine is diluent with phosphate buffer (pH=7) , then diluent samples extracted with Oasis WCX solid-phase extraction column. Samples were analyzed using LC-MS/MS in multiple reaction monitoring (MRM) mode. The analytical column was XBridge®BEH-HILIC (100 mm×2.1 mm×2.5 µm) and the mobile phase were 100 mmol ammonium formate add 0.5% formic acid and acetonitrile. Paraquat was quantified by internal standard method and diquat by external standard method. Results: The calibration curves of paraquat and diquat were linear in the concentration range of 10.0~120.0 µg/L, the correlation coefficient (r) were 0.9985~0.9994. The limit of detection of paraquat in plasma and urine were 1.98 µg/L and 1.00 µg/L, respectively, the recovery rate were 100.2%~107.3%, the RSD were 1.6%~3.3%. The limit of detection of diquat in plasma and urine were 1.80 µg/L and 2.77 µg/L, respectively, the recovery rate were 85.3%~93.1%, the RSD were 1.8%~5.5%. Conclusion: This method is sensitive and accurate, and can simultaneously determine paraquat and diquat in plasma and urine.

[The study on the detection method for mercury in blood with direct mercury analyzer].

Objective: To establish a method for determining mercury in blood with direct mercury analyzer. Methods: After the whole blood sample was extracted by solvent and removed by nitric acid, it was then measured by direct mercury analyzer. Results: After optimizing the conditions of the instrument, the linear range was 0.3-60.0 µg/L and the curve correlation coefficient was higher than 0.999. The lower limit of quantitations was 0.3 µg/L and the minimum quantitative concentration was 3.0 µg/L. The recovery and relative standard deviations (RSD) was 95.2%-97.6% and 1.4%-3.3%. Conclusion: The method is stable, reliable, easy to operate and has high sensitive. It can be used to determine mercury in blood.

[Elemental and isotopic determination of lead in whole blood by ISIS-ICP-MS].

Objective: To establish a method for determination of lead and istope ratios in the blood by ISIS-ICP-MS. Methods: After wet digestion, the blood sample was on-line addition of thallium as internal standard and analyzed by ISIS-ICP-MS. Results: The limit of detection was 0.03 µg/L and the lower limit of quantification was 0.08 µg/L. The detection concentration was 0.45 µg/L and the minimum quantitative concentration was 1.49 µg/L. The relative standard deviations (RSD) were 0.3%~1.7%. The recovery was between 91.0% and 103.4%. The precision of the major lead isotope ratios was better than 0.3%. The calibrated isotope ratios of the standard liquid are close to the certificate. Conclusion: The method has a low detection limit, good precision and high accuracy, it is feasible for determination of lead concentration and isotope ratios in the bloune.

[Determination of Sevoflurane, Isoflurane and Enflurane in the air of workplace by gas chromatography].

Objective: To establish a solvent desorption gas chromatographic method for determination of Sevoflurane, Isoflurane and Enflurane in the air of the Workplace. Methods: Sevoflurane, Isoflurane and Enflurane were collected with activated carbon tube and desorbed with dichloromethane, separated with DB-1 capillary columns, and then detected with flame ionization detector. Results: The linearity ranges were 1.9-304.8 µg/ml for Sevoflurane, 2.1-300.4 µg/ml for Isoflurane and 1.7-305.2 µg/ml for Enflurane, The correlation coefficient was both >0.999. Their limits of detection were 0.6 µg/ml, 0.6 µg/ml and 0.5 µg/ml, and Their limits of quatification were 1.9 µg/ml, 2.1 µg/ml and 1.7 µg/ml, and their minimum detectable concentrations were 0.1、0.2 and 0.1 mg/m(3) per 4.5 L of air. Their relative standard deviations (RSD) were 2.5%-3.0%, 2.3%-3.1% and 2.2%-3.0%. The average desorption efficiencies were 101.1%-103.3%, 100.7%-102.7% and 101.0%-102.9%. The sampling efficiency was both 100%. The breakthrough volume of 100 mg actived carbon was 3.7 mg, 3.4 mg and 3.4 mg. Sevoflurane, Isoflurane and Enflurane in activated carbon tube could be kept at least 10 days at room temperature without significant losses. Conclusion: The method shows lower detection limit, high accuracy and precision. It is feasible for determination of Sevoflurane, Isoflurane and Enflurane in the air of workplace.

[Simultaneous determination of 12 rodenticides in whole blood and urine samples by high performance liquid chromatography-tandemmass spectrometry].

Objective: To develop a method for Simultaneous and rapid determination of 12 rodenticides including pindone, vacor, coumatetralyl, warfarin, diphacinone, coumachlor, chlorphacinon, difenacoum, brodifacoum, bromadiolone, difethialone and flocoumafen in whole blood and urine samples by high performance liquid chromatography-tandem mass (LC-MS-MS) . Methods: The whole blood samples were precipitated with acetonitrile, purified by OstroTM 96-well plate, The urine samples were extracted by acetonitrile, and then separated on a ODS column, analyzed with high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) . The external standard calibration were tested. Results: A good linearity was observed in their respective concentration ranges of 12 rodenticides. The related coefficients were 0.993 0~0.999 8. The limit of detections were 0.05 µg/L~1.4 µg/L. The rates of recovery were 92.5%~118.0%. The relative standard deviations were between 0.8%~17.3%. Conclusion: The method was simple, rapid, sensitive, accurate and suitable for simultaneous detection of the 12 rodenticides in whole blood and urine samples of intoxicated patients.

[Direct determination of manganese in urine by Graphite Furnace Atomic Absorption Spectrometry].

Objective: To establish a method for the determination of manganese in urine by graphite furnace atomic absorption spectrometry (AAS) without the use of matrix modifier. Methods: The urine samples were 5 times diluted with 1% nitric acid then directly determined by AAS. Zeeman was used for background correction. Results: The linear range for determination of manganese in urine was 5~60 µg/L (urine) . The correlation coefficient was greater than 0.995 with the detection limit of 1.5 µg/L and with the lower limit of quantification of 5.0 µg/L. The relative standard deviations (RSDs) of within-run precision was between 1.1%~4.3%, the RSDs of between-run precision was between 3.3%~7.0%. The average recovery was 102.6%. The samples can be stored for 14 days at room temperature, 4℃, -8 ℃ and -35 ℃. Conclusion: The method is feasible for determination of manganese in urine.

[Simultaneous determination of various volatile organic compounds in urine by headspace GC-MS].

Objective: To establish a method for determination of acetone, dichloromethane, hexane, 1, 1, 1-trichloroethane, 1, 2-dichloroethane, benzene, toluene, ethylbenzene etc organic compounds in urine by headspace gas chromatography-mass spectrometry (GC-MS) . Methods: Headspace gases of urine samples were injected into GC and determined by mass. Results: Determination of urine components were in a good linear range in their concentration range of this method. The correlation coefficients were between 0.996 and 1.000 with the detection limits between 0.1 µg/L and 4.5 µg/L, the precisions were between 1.3% and 4.6%, the recovery rates were between 86.2% and 97.4%. Conclusion: This method has the advantages of low detection limits, high accuracy, high precision and simple pretreatment, which is suitable for the determination of the content of various volatile organic compounds in urine.

[Determination of methylenedianiline in urine by high performance liquid chromatography-tandemmass spectrometry].

Objective: To develop a method for determination of metabolites of diphenylmethane diisocyanate (MDI) in urine, i.e. methylenedianiline (MDA) by high performance liquid chromatography-tandem mass (LC-MS-MS) . Methods: Urine samples were prepared by hydrolyzation with sulfuric acid and extraction by acetonitrile, and then separated on a Shim-pack XR-ODS column, analyzed with high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) . The external solvent standard calibration were tested. Results: The linearity ranges were 0.05~20.00 µg/L, The related coefficients were 0.999 5. The limit of detection was 0.02 µg/L. The rats of recovery were 91.0%~103.4%. The relative standard deviations were between 2.7%~7.3%. Conclusion: The method was sensitive, accurate and suitable for the MDA determination in urine of MDI exposed population.

Baseline antibody level may help predict the risk of active human cytomegalovirus infection in a HCMV seropositive population.

Recurrent human cytomegalovirus (HCMV) infection during pregnancy could lead to congenital HCMV infection and permanent sequelae. However, knowledge regarding the risk factors associated with recurrent HCMV infection is limited. In the present study, 1,659 paired serum samples from the natural population were collected in Guangxi Province, China, from 2003 to 2004 with a 1-year interval. The serum anti-pp150 titre was quantitatively determined using a homemade recombinant pp150-based ELISA, and the IgG titre that increased at least 4-fold was defined as a recurrent infection. The HCMV seroprevalence was above 98.6% (1,636/1,659) in Guangxi in 2003, and the infection rate during the 1-year follow-up was approximately 10% (171/1,659). The seronegative population has the highest infection risk, while the risk of recurrent infection in the seropositive population was negatively correlated with the baseline anti-pp150 titre. With a cutoff of 1:80 (the baseline anti-pp150 IgG titre), the sensitivity and specificity were 73.1% (125/171) and 85.7% (1,275/1,488) respectively, and the relative risk of infection in the high-risk group compared to the low-risk group was 10.6 (95% CI: 7.7-14.6). In conclusion, the baseline anti-pp150 IgG was negatively correlated with the risk of HCMV infection and could be an excellent predictor of HCMV infection in HCMV seropositive populations.

[Determination of tert-butyl alcohol in the air of the workplace by solvent desorption gas chromatography].

Objective: To establish solvent desorption gas chromatographic method for determination of tert-butyl alcohol in the air of the workplace. Methods: After tert-butyl alcohol in the air of the workplace collected with activated carbon tube and desorbed with 2% 2-propanol in CS2, and then separated with DB-FFAP capillary column and detected with flame ionization detector. Results: The linearity ranges were 0.6~2 264.0 mg/L. The limit of quantification was 0.6 mg/L. The determination has a good reproducibility. The intraassay and interassay precisions were 2.8%~3.2% and 3.8%~5.7%. The desorption efficiencies were 93.9%~98.1%. Absorption efficiencies were 95.8%~100.0%. The breakthrough volume was above 7.1 mg in 100mg activated carbon. The samples in activated carbon tube could be stored for at least 14 days at ambient temperature. Conclusion: The method is feasible for determination of tert-butyl alcohol in the air of the workplace.

Tempero-spatial dissociation between the expression of Fas and apoptosis after coronary occlusion.

AIMS: To explore the role of Fas in cardiomyocytic apoptosis induced by ischaemia through determining the histological relation between Fas expression and apoptosis in rat myocardium during ischaemia/infarction. METHODS: The myocardial ischaemia model was produced by ligating the left coronary artery in Sprague-Dawley rats. The rats were killed from 10 minutes to seven days after surgery. Apoptotic myocardial cells were detected by the in situ terminal deoxynucleotidyl transferase mediated nick end labelling method, and the expression of Fas by immunohistochemistry and western blotting. RESULTS: Cardiomyocytic apoptosis appeared from three to 36 hours after ischaemia. The expression of Fas could be detected by western blot from before surgery to seven days of ischaemia. Apoptosis and the expression of Fas in the cardiomyocytes appeared in different regions of the myocardium: apoptosis in the ischaemic region, Fas in the regions surrounding ischaemic myocardium. CONCLUSION: These results suggest that there is a tempero-spatial dissociation between the expression of Fas and apoptosis after coronary occlusion. Fas might not directly regulate the apoptosis of cardiomyocytes induced by ischaemia.

The role of xanthine oxidase and xanthine dehydrogenase in skin ischemia.

The importance of sequential events which lead to skin necrosis has significant implications in trauma, vascular injury, and wound healing. In this series of experiments, we tested the hypothesis that xanthine oxidase (XO) activity was increased along an ischemic gradient of a skin flap and that the XO enzyme activity correlated with an increase in neutrophils. There were two animal groups in which the skin flaps were raised and assayed at 0, 1, or 6 hr. In the other group, they were created as bipedicle flaps for 7 days, before the distal attachment was divided and the tissue assayed. In the acutely raised flaps, some animals were treated with the XO inhibitor, allopurinol. Xanthine dehydrogenase (XD) and XO activity was measured with a fluorometric pterin assay and neutrophil concentration was measured using a myeloperoxidase marker. In this model, there was consistent skin necrosis in the distal end of the skin flap (48 +/- 8%). The data showed that both XD and XO activity in the distal ends was statistically significantly increased over the sham control or proximal ends of the skin flaps at 1 hr (P < 0.05). XO activity remained elevated in the distal ends at 6 hr. Allopurinol significantly reduced the neutrophil concentrations in the distal ends of the skin flaps when compared to untreated animals (P < 0.05). Moreover, allopurinol reduced skin necrosis to 12 +/- 1%. Preconditioning of the skin flap reduced the XO activity to sham control levels. The observations implicate XO activity as source of free radical injury in skin necrosis seen in random skin flaps.(ABSTRACT TRUNCATED AT 250 WORDS)

[Comparison of lipoperoxidative damage in plasma and platelet with the ratio of thromboxane A2 and prostaglandin I2 in blood stasis syndrome with coronary heart disease].

In order to study the biochemical and pathophysiological mechanism of the Blood Stasis Syndrome (BSS) or Non-BSS of coronary heart disease (CHD) patients, the activities of SOD, Selenium-glutathione peroxidase, the content of LPO in plasma and platelets and the contents of TXB2 and 6-keto-PGF1 alpha in plasma were determined in 109 BSS and Non-BSS of CHD patients compared with 98 healthy controls. It was discovered that the contents of TXB2, LPO, PL-LPO, and the ratio of TXB2/6-keto-PGF1 alpha were significantly increased in BSS-CHD patients compared with controls and Non-BSS-CHD patients. It was also discovered that the SOD activities and the contents of 6-keto-PGF1 alpha decreased significantly in Non-BSS-CHD patients. The results suggested that the injury of platelets by oxygen free radicals might be one of the primary injury factors in BSS-CHD patients. Our conclusion is that PGI2, SOD belong to the category of Heart-Qi, while TXA2, LPO to the Blood category. Therefore TXB2, 6-keto-PGF1 alpha, SOD, LPO should serve as some of the objective indexes for BSS patients of CHD.

Defibrase, a purified fibrinolytic protease from snake venom in acute myocardial infarction.

To investigate the thrombolytic effects of defibrase in AMI, 157 pts with AMI were studied. Of the 157 pts, 87 were assigned to defibrase thrombolysis and 70 to the conventional therapy plus heparin (control). Coronary arteriography was performed in 36 pts of the defibrase group and 26 of the control group. Pretreatment coronary arteriography was performed in 10 pts and total occlusion of infarct-related artery (IRA) was demonstrated in 7. Thirty to 45 min after either intracoronary or intravenous defibrase thrombolysis, recanalization occurred in 4 of the 7 pts. The perfusion and stenosis was improved in 2 of the 3 pts with patent IRA. Of the 10 pts who underwent emergency coronary arteriography after the onset of i.v. defibrase thrombolysis, 6 were shown to have patent IRAs. Of the 16 pts who underwent coronary arteriography two weeks after the i.v. defibrase thrombolysis, 13 were shown to have patent IRAs. In contrast, only 11 of the 26 pts in the control group had patent IRAs two weeks after admission to the hospital. Of the 36 angiographic cases of the defibrase group, 15 underwent follow-up coronary arteriography, only 1 pt showed reocclusion. In comparison to the control group, the pts in the defibrase group had earlier CPK peaking, higher percentage of pts with LVEF > 0.5, a lower mortality and complication rate. Major spontaneous bleeding complications were rarely seen with defibrase. The results indicate that defibrase is an effective thrombolytic agent with a reasonable recanalization rate, low reocclusion rate and a low rate of bleeding complications.

[Coronary thrombolysis with defibrase].

Clinical use of defibrase (DF), a fibrinolytic agent from venom of Agkistrodon acutus, was investigated in patients with acute myocardial infarction (AMI). Patients with AMI were randomized to tow groups, one receiving conventional therapy and DF, the other a control group having conventional treatment only. The patients in the DF group received intravenous DF 0.05 u or 0.075 u/kg over 1 hour immediately after the attack and 0.025 u/kg over 4 hours on the 5 th and 10 th day. The control patients received conventional therapy only. Of the patients randomized, 21 had coronary angiography and left ventriculography. Recanalization was seen in 10 of the 10 patients treated with DF, while only in 3 of the 11 patients in the control group. LVEF in the DF group was higher than that in the control group (61% vs 50%). However the difference was not significant. In the DF group, two patients developed petechia and blood oozing at the site of intravenous injection and one patient developed gum bleeding; all patients had transient thrombocytopnia, which returned to normal within 5 days. There was no intracranial hemorrhage, gross hematuria or hematemesis. In conclusion, DF possess thrombolytic as well as anticoagulant effects with minor bleeding complication. Early infusion of DF yields high patency rate and shows a trend toward preservation of LV function.

[Smooth muscle tumors of the alimentary tract--a review of 397 cases].

397 cases of smooth muscle tumor of the alimentary tract were analysed. 385 were collected from the domestic literature in the recent 10 years and 12 are added by the authors. The tumor occurred more frequently in patients 40-60 years old. Diameter of the tumor ranged from 0.5-35 cm. The diameter of leiomyosarcoma was larger than that of leiomyoma on an average. The ratio of benign to malignant incidence was 3.2:1. Exclusive of 204 (51.4%) cases which occurred in the esophagus, the ratio was 1.1:1 in the other 193. In the 12 cases reported by the authors, it was 1:3. In the 193 cases of the GI tract, the lesion occurred in the stomach in 50.8%, in the small intestine in 29.0%, in the colon 1.0% and in the rectum 19.2%. The three most common clinical symptoms of those in the stomach and small intestine were abdominal mass (54.3%), bleeding (49.1%) and abdominal pain (45.7%). These 12 cases reported comprised 0.019% of 62, 088 patients admitted into our hospital, 0.09% of 12,678 tumor inpatients during the same period and 0.61% of 1,983 of malignant tumors in the alimentary tract. The preoperative diagnostic accuracy of this tumor was low, only 7.7% of all the GI neoplasms. On basis of the present data, improving the preoperative diagnostic accuracy is stressed.

[Effect of pineal body extract on cytoxan-induced leucopenia in mice].

The effect of pig pineal extract on cytoxan-induced leucopenia in mice was studied. Three preparations of pineal body, Songlesu A, B and C, were used. All of them were effective in leucopenia. They delayed the time of the lowest white count, reduced the extent of white cell reduction and elevated and sustained the white count for several days despite the continuation of cytoxan injection. The effect of Songlesu A was most marked. The differences between the white count of group A (Songlesu A plus cytoxan) and the control (cytoxan plus normal saline) are very significant (P less than 0.01) in a period of 8-15 days after giving the drugs. After discontinuation of medication, the recovery of group A was better than that in the control group also very significantly (P less than 0.01). It is evident that Songlesu A is most worthy of further investigation. However, the effect of Songlesu A was no longer marked from the seventeenth day when the white count started to decline. This is most probably due to the severe bone marrow damage by the large dose of cytoxan accumulated or the protracted daily administration. Moreover, Songlesu A may sensitize the bone marrow to cytoxan toxicity or change the rate of recovery after bone marrow damage.