ObjectiveTo establish an accurate method for simultaneous determination of plasma Kyn and Trp by HPLC-UV detection.Methods Kyn and Trp were separated on Agilent Hypersil ODS column using 3-nitrotyrosine as internal standard.The mobile phase consisted of 15 mmol/L sodium acetateacetic acid (pH 5.5):acetonitrile 94∶ 6(v/v) at a rate of 0.8 ml/min.The chromatographic separation was performed at 25 ℃.The eluate was monitored with programmed wavelength setting at 360 nm from 0 to 4 min for Kyn and at 302 nm from 4 to 5 min for Trp.The method was applied to determination of plasma Kyn and Trp in 8 chronic glomerulonephritis,10 idiopathic thrombocytopenic purpura,15 chronic hepatitis B virus patients and 15 healthy controls from September to December in 2010.The differences were compared using ANOVA and SNK methods.Results The retention time of Kyn and Trp were 2.9 min and 4.4 min,respectively.For Kyn,the assay was linear from 0.44 μmol/L to 18.30 μmol/L.For Trp,the linearity was from 3.67 μmol/L to 470.00 μmol/L.The detection limits were 0.014 μmol/L for Kyn and 0.122 μmol/L for Trp,respectively.The within-day CVs were ＜ 3％ and the between-day CVs were ＜ 4％.The mean recoveries yield were in the range of 92.29 to 104.40.The plasma concentrations of Kyn were ( 1.59 ± 0.28),(2.73 ± 0.56),(2.69 ± 0.44) and ( 1.54 ± 0.48 ) μmol/L,the plasma concentrations of Trp were (59.8 ± 10.0),(46.1 ± 11.7),(58.5 ±8.0) and (41.4±13.1) μmol/L,the Kyn/Trp were (0.027 4±0.007 5),(0.061 6 ±0.016 5),(0.046 7 ±0.009 1) and (0.038 3 ±0.007 5)in controls,chronic glomerulonephritis patients,idiopathic thrombocytopenic purpura patients and chronic hepatitis B virus patients,respectively.There were significance difference of Kyn,Trp and Kyn/Trp amony the four groups (F=23.734,8.463,20.921,all P＜0.01).Conclusion The method is simple,fast,and suitable for applicability to clinical measurement.

Objective:Aimed to investigate the value of adrenocorticotropic hormone (ACTH) stimulation in adrenal venous blood sampling (AVS).Methods:Patients who diagnosed as primary aldosteronism (PA) and completed successful bilateral cannulation judged by selection index (SI) for routine and(or) ACTH stimulation AVS were enrolled. The lateralization index(LI) was calculated to compare the effect of ACTH stimulation on AVS cannulation success rate and lateralization judgment.Results:A total of 73 patients with PA were enrolled in the study, of whom 28 were confirmed as aldosterone producing adenoma (APA) after unilateral adrenalectomy. Cortisol and aldosterone in peripheral and adrenal veins were significantly increased after ACTH stimulation. The left SI was increased from 6.5(3.0-13.6) to 26.8 (16.9-40.3) ( P<0.01) and the right SI from 20.8(4.8-34.8) to 57.6(35.7-80.9) ( P<0.01) after ACTH stimulation. There was no significant difference on LI before and after ACTH stimulation [7.7(2.3-19.6) vs 5.6(1.9-14.6), P=0.14]. The success rates of left and right adrenal cannulation were increased by 15% and 10% respectively after ACTH stimulation. For 57 patients who were determined in successful cannulation by both routine and ACTH stimulation AVS, 27 patients were determined to have lateralization by both AVS methods, 21 patients were determined to have bilateralization, and the consistency of lateralization by both AVS methods was 84%(48/57). Among the 28 patients who were confirmed to be APA after unilateral adrenalectomy, the correct rate of lateralization by both AVS methods was 89% (25/28). Conclusion:ACTH stimulation is able to improve the success rate of bilateral adrenal vein cannulation, and is helpful to judge AVS results. For patients with successful cannulation, there is no significant difference in lateralization judgment for routine and ACTH stimulation AVS.

Objective To assess the diagnostic value of saline infusion test ( SIT) and its optimal cutoff in the diagnosis of primary aldosteronism ( PA ), and to analyze whether the dietary salt intake affects the SIT accuracy. Methods This is a prospective study. All 236 patients with a high risk for PA underwent the screening test, SIT and the fludrocortisone suppression test (FST) in separate days. The diagnosis of PA was established according to the FST criteria. According to the 24 h urinary sodium level, the patients were divided into low salt, normal salt, and high salt groups respectively, and the effect of salt intake on SIT was analyzed. Receiver operating characteristic (ROC) analysis was performed to compare the diagnostic accuracies. Results Finally, in 236 patients with high risk for PA, 134 patients with PA and 102 patients with essential hypertension ( EH) were diagnosed. Using post-test plasma aldosterone concentration (PAC) for diagnosis, the area under the ROC curve (AUCROC) of the SIT was 0.974 (0.957, 0.991), which was significantly higher than that of the post-test plasma aldosterone to renin ratio (ARR) [0.900 ( 0. 862, 0. 938)] and that of the PAC suppression percentage [ 0. 752 ( 0. 690, 0. 813)] ( both P<0.01). Considering both sensitivity and specificity, an optimal cutoff of PAC post-SIT was set at 8 ng/dl, resulting in a sensitivity of 88. 1% and a specificity of 95. 1%. The PAC post-SIT, whether in PA or EH patients, had no statistically significant differences among low salt, normal salt, and high salt groups (P>0.05). Conclusion SIT is reliable for the diagnosis of PA. PAC post-SIT more than 8.0 ng/dl is recommended to confirm PA.

Objective To investigate the tryptophan (Trp) metabolism change in the patients with chronic hepatitis B (CHB) and hepatitis B cirrhosis (HBC) and to analyze the related risk .Methods Eighty-one cases of CHB and 40 cases of HBC hospitalized in the First Affiliated Hospital of Chongqing Medical University from October 2015 to January 2016 served as the stud-y subjects ,and 40 healthy persons as the controls .High performance liquid chromatography -fluorescence detection (HPLC-FD) were adopted to detect the levels of plasma Trp ,kynurenine (Kyp) and its metabolite 5-hydroxy tryptamine (5-HT) .HBV-DNA vi-ral replication number was detected by real-time quantitative PCR .The serological indicators differences among 3 groups were ana-lyzed .The risk factors were analyzed and screened by the Logistic regression analysis and Pearson correlation analysis .Results Compared with the control group ,plasma 5-HT and Trp levels in the CHB group and HBC group were significantly decreased (P<0 .01);plasma Kyn/Trp in the HBC group was significantly higher than that in the other two groups (P<0 .01) .Kyn/Trp in the HBC group was negatively correlated with A/G (r= -0 .686 ,P<0 .01) ,and Kyn/Trp in the CHB group was positively correlated with Log DNA (r= 0 .784 ,P<0 .01) .The Logistic regression analysis indicated that Kyn/Trp was the risk factor of CHB and HBC .Conclusion Detecting plasma Trp and its metabolite can serve as the adjuvant indicators for assessing the progression of hep-atitis B .

Objective:To investigate the distribution of pathological types of unilateral primary aldosteronism, and to explore the clinical characteristics and prognosis of patients with different pathological types.Methods:A total of 241 patients with unilateral primary aldosteronism who underwent adrenal surgery were included in this study. The clinical data and postoperative follow-up data were collected, and the postoperative tissue sections were stained with HE and aldosterone synthase. According to the staining results, pathological types of 241 patients were classified, and the clinical characteristics and surgical prognosis of patients with unilateral primary aldosteronism were compared.Results:According to the international histopathology consensus for unilateral primary aldosteronism, among 241 patients with unilateral primary aldosteronism, 223 were classical(92.5%), 17 were non-classical(7.1%), and 1 was aldosterone producing carcinoma(0.4%). Among classical cases, 189 were aldosterone producing adenoma and 34 were aldosterone producing nodule. In the non-classical cases, 8 cases were multiple aldosterone producing nodule and 9 cases were multiple aldosterone producing nodule. Compared with the classical group, the non-classical group had a longer duration of hypertension(9.0 vs 5.0 years, P=0.062) and a lower baseline plasma aldosterone concentration(273 vs 305 pg/mL, P=0.147), but the difference was not significant. There was no significant difference between the two groups in the proportion of patients who achieved a complete biochemical response after surgery(98% vs 92.3%, P=0.281), but the proportion of patients who achieved a complete clinical response was significantly lower in the non-classical group(23.1% vs 52.9%, P=0.046). Conclusion:The pathological types of unilateral primary aldosteronism are predominantly classical, with aldosterone-producing adenoma being the most common. There were no significant differences in the clinical characteristics and postoperative biochemical remission rates between classical and non-classical patients, but the clinical prognosis of the latter was inferior to the former.

To investigate the renal venous and spermatic venous sampling in assisting the diagnosis of reninoma. The case of reninoma was retrospectively reviewed together with the literature review of reninoma diagnosed with renal venous and spermatic venous sampling. A young patient with hypertension and headache was admitted to our hospital. Laboratory test showed high plasma renin concentration (>500 mIU/L), and enhanced computed tomography(CT) in the upper abdomen showed a mass in left inferior renal pole. The concentration of renin in the left spermatic vein was significantly higher than that in renal veins and branches, and peripheral vein, which was considered the left reninoma possibility. The left renal mass was resected surgically and pathologic exam revealed reninoma. The renin level and the blood pressure recoveried normal after operation.The literature reported that the positive rate of renal vein segmental blood collection for locating renin tumor was only 8.3%-64%. The possible reason was that reninoma usually located in the renal cortex, and the tumor blood might be collected by renal capsule vein instead of renal vein. In fact, the renal capsule vein intersects with the lateral division of the spermatic vein, but there have been no reports about the localization of reninoma by spermatic vein sampling. Since renin secreted by reninoma may go into the spermatic vein through renal capsule vein, it should be noted that spermatic venous blood should be collected simultaneously in renal vein sampling when locating reninoma.

Objective: To compare the detection efficiency of the ratio of plasma aldosterone concentration to renin concentration (ARR) measured by imported and domestic chemiluminescent immunoassay kits for screening primary aldosteronism (PA) . Methods: A total of 164 patients with essential hypertension and 64 patients with PA were recruited in this study. Plasma aldosterone concentration (PAC) and renin concentration (PRC) were measured concurrently by imported chemiluminescent immunoassay kit (Diasorin, Italy) and domestic kit (Mindray, Shenzhen, China) , and then ARR were calculated. A ROC curve analysis was performed to compare the screening efficacy of the two kits and the optimal ARR cut-offs for PA screening were recommended according to Youden’s index. Results: The areas under the ROC curves (AUCROC) of ARR were 0.937 (95% CI 0.897-0.965, P<0.0001) detected by imported kit and 0.942 (95% CI 0.903-0.968, P<0.0001) detected by domestic kit. The difference of AUCROC of ARR detected by two kits was 0.00457 (95% CI -0.0210-0.0302, P<0.7263) .The highest Youden’s indexes of ARR were 0.749 (detected by imported kit) with the cut-off value of 12.8 (pg/ml) / (μIU/ml) (sensitivity 96.87%, specificity 78.05%) and 0.756 (detected by domestic kit) with the cut-off value of 12.5 (pg/ml) / (μIU/ml) (sensitivity 89.06%, specificity 86.59%) . Conclusions There was no significant difference between domestic and imported chemiluminescent immunoassays in the screening efficiency of ARR for PA.