1.A preliminary study on relationship between single nucleotide polymorphism ofmiR-196a2 and risk of leukemia
Wenjuan ZHANG ; Zhe TIAN ; Youjie LI ; Wenjing SHANG ; Ning XIE ; Jing CHANG ; Jing XIAO ; Xuelin BAO
China Oncology 2016;26(10):826-830
Background and purpose:miR-196a2 functions as an oncogene during tumor initiation and pro-gression. The up-regulation promotes tumor cell proliferation, invasion and metastasis. Therefore, it is promising to be an important tumor biomarker. The aim of this study was to investigate whether rs11614913, a gene polymorphic site ofmiR-196a2, is associated with the risk of leukemia.Methods:A case-control analysis was employed. Bone marrow or periph-eral blood was collected from 210 leukemia patients diagnosed from Jan. 2009 to Jul. 2015 in Yantaishan Hospital (case group) as well as 250 healthy people who were physically examined during the same period (control group). Polymerase chain reaction-restriction fragment length polymorphism (PCR-PFLP) was used to detect the genotype of rs11614913. Application test was used to compare the difference in the frequency of each genotype between case group and control group. The odds ratio (OR) of SNP allelic genes was calculated using logistic regression analysis and 95%CI represented the risk of leukemia for each genotype.Results:The distribution differences in the frequency of T/T, C/C, C/T genotype of miR-196a2 rs11614913 between case group and control group were statistically significant (P<0.05). The risk of leukemia for individuals who carried mutant homozygous C/C was 2.661-fold higher than those carried wild-type homozygous T/T, and the difference was statistically significant (P<0.05).Conclusion:ThemiR-196a2 gene polymorphic site rs11614913 was associated with the risk of leukemia. Mutant homozygous C/C or C allelic gene carrying was probably a risk factor for leukemia.
2.Correlation study between Mycobacterium tuberculosis PhoPR two-component system and drug resist-ance of widespread clinical isolates of Mycobacterium tuberculosis
Wenjuan LI ; Yin BAO ; Xia WANG ; Feng ZHANG ; Chen LIANG ; Rui ZHUANG ; Chao FAN ; Qingzhang TUO ; Chunjun ZHANG ; Hui ZHANG ; Fang WU ; Jiangdong WU ; Le ZHANG ; Wanjiang ZHANG
Chinese Journal of Microbiology and Immunology 2013;(10):773-779
Objective To explore the correlation between Mycobacterium tuberculosis ( MTB ) PhoPR two-component system and drug resistance of MTB clinical isolates widespread in Xinjiang region by analyzing the expression of PhoP gene and PhoR gene among different isolates .Methods Total RNA of MTB was extracted from drug-susceptible strains , the strains only resistant to a single first-line anti-TB drugs (INH, RFP, SM and EB) and multidrug-resistant (MDR) strains, respectively.The purity of total RNA was checked by agarose gel electrophoresis .The expressions of PhoP gene and PhoR gene were quantified by using SYBR Green I qRT-PCR and the differences of their gene expression in different isolates were ana-lyzed.Results Compared with the drug-susceptible strains of MTB, the expression of PhoP gene was up-regulated for about 1.48 times in MTB strains resistant to RFP (RFP-MTB) and 2.74 times in MDR strain (P<0.05).Compared with MDR strain, the expressions of PhoP gene in the isolates resistant to INH (IN-HMTB), RFP (RFP-MTB), SM (SM-MTB) and EB (EB-MTB) were down-regulated for 0.70, 0.50, 0.25 and 0.21 times respectively.The expressions of PhoR gene were down-regulated for 0.36, 0.54, 0.35 and 0.19 times, respectively (P<0.05).The expressions of PhoR gene in the isolates of INH-MTB, RFP-MTB, SM-MTB and EB-MTB were up-regulated for 6.33, 4.56, 2.34, 1.85 and 9.06 times, respectively as compared with the drug-susceptible strains (P<0.05).Conclusion Significant differences of PhoR gene and PhoP gene expressions were observed among drug-susceptible strains , INH-MTB, RFP-MTB, SM-MTB, EB-MTB and MDR strains.Therefore, the Mycobacterium tuberculosis PhoPR two-component system is asso-ciated with the drug resistance of MTB strains prevalent in Xinjiang region .
3.Effect on expression of macrophages ferroportin and ferritin in mouse alveolar macrophages by mycobacterium tuberculosis
Rui ZHUANG ; Xia WANG ; Feng ZHANG ; Yin BAO ; Le ZHANG ; Fang WU ; Jiangdong WU ; Chunjun ZHANG ; Hui ZHANG ; Wenjuan LI ; Chen LIANG ; Chao FAN ; Wanjiang ZHANG
Chinese Journal of Immunology 2014;(5):591-595
Objective:To discuss the change of ferritin ( Fn) and ferroportin expression quantity and time-related feature in the alveolar macrophages of mice , infected with different virulence of Mycobacterium Tuberculosis infected .Methods:The prepared bacte-ria of H37Rv or BCG were injected intravenously into the mice tails .On the day 1, 3, 5, 7, 9, 11, 13 and 15, the lavage fluids were collected and the alveolar macrophages were obtained from each group of mice .The expression of FPN and Fn were detected with ELISA and /or Western blot analysis .Results:The expression of Fn in the group of either H 37Rv or BCG infected mice was decreased on the day 7, 9 and 11, and was lowest on the day 7, which showed significantly statistical difference compared to that on the other days (P<0.05).The expression of FNP in the infected mouse macrophage was decreased gradually , which was obvious on the day 5. The expression levels reached to the lowest on the day 7 and 9.The expression was much lower than that in the negative control group (P<0.05).Conclusion:The expression of Fn and FPN in macrophages isolated from lungs of mice infected with Mycobacterium tu -berculosis H37Rv or BCG become decreased , and there is no difference between these two infected mouse groups .
4.Comparison of different optimized anesthesia strategies in pediatric patients undergoing hypospadias surgery
Yi GAO ; Pengqi DAI ; Lei SHI ; Wenjing CHEN ; Wenjuan BAO ; Lanlan HE ; Yongpan TAN
Chinese Journal of Anesthesiology 2020;40(2):190-194
Objective:To compare the different optimized anesthesia strategies in pediatric patients undergoing hypospadias surgery.Methods:Ninety pediatric patients with distal hypospadias undergoing Duckett operation, of American Society of Anesthesiologists physical status Ⅰor Ⅱ, aged 1-6 yr, weighing 10-25 kg, were divided into 3 groups ( n=30 each) using a random number table method: caudal block combined with general anesthesia group (group CG), neurostimulator-guided pudendal nerve block combined with general anesthesia group (group PG) and ultrasound-guided dorsal penile nerve block combined with general anesthesia group (group DPG). Anesthesia was induced with propofol-fentanyl, patients were ventilated through the laryngeal mask and maintained spontaneous breathing, and anesthesia was maintained with sevoflurane.The corresponding regional block was performed in each group, and block was performed with 0.25% ropivacaine 1, 0.25 and 0.15 ml/kg in CG, PG and DPG groups, respectively.The consumption of ropivacaine, occurrence of insufficient analgesia during operation, emergence time, duration of post-anesthesia care unit stay, time of the first requirement for analgesics, requirement for analgesics within 24 h after operation, occurrence of postoperative agitation, nausea and vomiting and lower extremity motor block, and patients′ satisfaction score at 24 h after operation were recorded. Results:Compared with group CG, the consumption of ropivacaine was significantly reduced, the time of the first requirement for analgesics was prolonged, the postoperative requirement for analgesics was decreased, duration of post-anesthesia care unit stay was shortened, the incidence of lower extremity motor block was decreased, and patients′ satisfaction score was increased in group PG and group DPG, and the incidence of nausea and vomiting was decreased in group DPG ( P<0.05). Compared with group PG, the consumption of ropivacaine was significantly reduced, and the time of the first requirement for analgesics was shortened in group DPG ( P<0.05). There was no significant difference in the incidence of insufficient analgesia during operation (3% in group DPG) and incidence of postoperative agitation among the three groups ( P>0.05). Conclusion:Ultrasound-guided dorsal penile nerve block combined with general anesthesia provides significant efficacy with a higher safety, and the efficacy is better than that of caudal block combined with general anesthesia and neurostimulator-guided pudendal nerve block general anesthesia when used for pediatric patients undergoing hypospadias surgery.
5.PD-1 is associated with CD8 +T lymphocyte dysfunction in patients with acute and chronic liver failure
Le CHANG ; Xin ZHANG ; Yanping ZHANG ; Dongyuan QIN ; Wenjuan LIU ; Bao CHAI ; Jia YAO
Chinese Journal of Hepatology 2021;29(11):1101-1105
Objective:To explore whether peripheral blood CD8 +T lymphocyte dysfunction is correlated with the programmed death receptor-1 (PD-1) expression in patients with acute-on-chronic liver failure (HBV-ACLF). Methods:Peripheral blood mononuclear cells (PBMC) were collected from patients with HBV-ACLF and healthy controls. CD8 +T lymphocytes number and PD-1 expression condition in CD8 +T lymphocytes were detected by flow cytometry. CD8 +T lymphocytes isolated from peripheral blood of HBV-ACLF patients were further cultured in vitro. One group was added with PD-L1-IgG fusion protein (ACLF+PD-1 group), and the other group was added with IgG fusion protein (ACLF group). Proliferation ability (ki67), cell viability (CD69), and secretion ability of effector cytokines (IL-2, IFN-γ, TNF-α) were analyzed. Results:30 cases with HBV-ACLF and healthy controls were enrolled. CD8 +T lymphocytes absolute number was significantly lower in the peripheral blood of patients with ACLF group (333.88 ± 147.74)/μl than healthy controls (872.50 ± 206.64)/μl ( P < 0.001). PD-1 expression in peripheral blood CD8 +T lymphocytes were significantly increased in ACLF group (13.33% ± 2.52%), ( P = 0.027) than healthy controls (7.02% ± 2.12%). In in vitro culture, compared with healthy controls, the peripheral blood CD8 +T lymphocytes cell viability (CD69), proliferation ability (ki67) (all P ??< 0.001), and the level of cytokine production (IL-2, IFN-γ, TNF-α) (all P < 0.05) were equally weakened in patients with ACLF group. Compared with ACLF group, CD8 +T cell viability (CD69), proliferation ability (KI67) (all P < 0.05), and the level of cytokine production were weakened in ACLF+PD-1 group (all P < 0.05). Conclusion:HBV-ACLF patients have CD8 +T lymphocyte dysfunction. Therefore, PD-1 may have correlation in the regulation of CD8 +T lymphocyte dysfunction in ACLF patients.
6.Effects of preoperative home-based rehabilitation in patients undergoing lung cancer surgery
Xiaoxia CHEN ; Zirui DING ; Lei HE ; Lijuan BAN ; Wenjuan ZHU ; Qingzhu BAO
Chinese Journal of Modern Nursing 2023;29(18):2482-2487
Objective:To explore the effect of preoperative home-based rehabilitation in patients undergoing lung cancer surgery.Methods:From January 2021 to June 2022, convenience sampling was used to select 118 lung cancer patients who underwent surgical treatment at the Xinxiang First People's Hospital of Henan Province. The patients were randomly divided into an observation group and a control group, with 59 cases each. The control group received routine nursing, while the observation group received preoperative home-based rehabilitation. We compared the lung function, nutritional indicators, incidence of postoperative atelectasis and pulmonary infection, as well as the duration of thoracic closed drainage tube insertion and postoperative hospital stay between the two groups.Results:After intervention, the forced vital capacity (FVC), maximum ventilation volume (MVV), the percentage of forced expiratory volume in one second to the predicted value (FEV 1%), ventilate storage ratio (VSR), hemoglobin and serum albumin in the control group were lower than those before intervention, and the levels of FVC, MVV, FEV 1%, VSR, hemoglobin and serum albumin in the observation group after intervention were higher than those in the control group, with statistical differences ( P<0.05). The incidence of postoperative atelectasis and pulmonary infection in the observation group were lower than that in the control group, and the differences were statistically significant ( P<0.05). The observation group had shorter duration of thoracic closed drainage tube insertion and postoperative hospital stay compared to the control group, with statistically significant differences ( P<0.05) . Conclusions:Preoperative home-based rehabilitation can effectively maintain postoperative lung function and nutritional status of patients, reduce postoperative complications, shorten postoperative drainage tube insertion time and hospital stay, and facilitate postoperative recovery.
7.Analysis of factors influencing stone recurrence and establishment of risk prediction model after laparoscopic common bile duct exploration combined with laparoscopic cholecystectomy
Xiaoyang ZHANG ; Hanxiang YU ; Junye WEN ; Wenjuan BAO ; Xing XU ; Yige SHI
Chinese Journal of Hepatobiliary Surgery 2024;30(10):766-770
Objective:To explore the risk factors for stone recurrence after laparoscopic common bile duct exploration (LCBDE) combined with laparoscopic cholecystectomy (LC) and to develop a risk prediction model.Methods:Clinical data of 344 patients with bile duct stones who underwent LCBDE combined with LC at Hebei General Hospital from January 2016 to March 2022 were retrospectively analyzed, including 165 males and 179 females, aged (62.72±13.56) years old. Patients were divided into two groups based on whether stones recurred during the follow-up period: recurrence group ( n=37) and non-recurrence group ( n=307). Clinical data such as common bile duct diameter, stone size, number of stones and duration of T-tube drainage were collected from the patients. Logistic regression was used to analyze the risk factors for postoperative stone recurrence, and then developed a logistic regression model. The predictive efficacy of the model was assessed by the area under the receiver operating characteristic (ROC) curve, and the Hosmer-Lemeshow test. Results:The results of multifactorial logistic regression analysis showed that patients with ≥2 choledochal stones had a high risk of stone recurrence after LCBDE combined with LC ( OR=3.094, 95% CI: 1.069-8.954, P=0.037). In contrast, regular postoperative oral choleretic medication was a protective factor for stone recurrence after LCBDE combined with LC ( OR=0.160, 95% CI: 0.072-0.354, P=0.001). A logistic regression model, based on the number of common bile duct stones and regular postoperative oral choleretic medication, was developed to predict the recurrence of bile duct stones in patients who underwent LCBDE combined with LC. The area under the ROC curve for this model was found to be 0.821 (95% CI: 0.758-0.885). The Hosmer-Lemeshow test, χ 2=7.26, P=0.509, suggested that there is good agreement between the model's predicted probabilities and ideal probabilities. Conclusions:The number of stones (≥2) is an independent risk factor for stone recurrence after LCBDE combined with LC in patients with bile duct stones. Regular postoperative oral choleretic medication is a protective factor for stone recurrence after LCBDE combined with LC. Predictive models based on the number of choledochal stones and regular postoperative oral choleretic medication have better efficacy in predicting postoperative stone recurrence.
8.Application value of multiple imaging techniques in the MDCT combined with assessment of vascular invasion using a tumor-to-vessel contact computed tomography grading system in preoperative evaluation of pancreatic cancer
Fangming CHEN ; Wenjuan WU ; Lei ZHANG ; Zhuiyang ZHANG ; Jialing BAO ; Yongping ZHOU
Chinese Journal of Digestive Surgery 2018;17(7):752-758
Objective To investigate the application value of multiple imaging techniques in the multidetector computed tomography (MDCT) combined with assessment of vascular invasion using a tumor-to-vessel contact (TVC) computed tomography grading system in preoperative evaluation of pancreatic cancer.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 66 patients with pancreatic cancer who were admitted to the Wuxi No.2 People's Hospital of Nanjing Medical University between February 2012 and July 2017 were collected.Surgical results of 66 patients showed that tumors of 48 and 18 patients were respectively located in head or uncinate process of the pancreas and body and tail of pancreas.The 317 vessels of 66 patients were detected.Fifty patients underwent radical resection and 250 vessels were detected,resected tumor diameter was (2.7± 1.4) cm (range,1.3-7.7 cm);16 underwent palliative operation and 67 vessels were detected.Patients received enhanced scans of MDCT,and multiplanar reformatted (MPR),curved planar reconstructions (CPR),CT angiography (CTA),arterial and venous images of three-dimensional (3D) CT and negative-contrast CT cholangiopancreatography (nCTCP) were build.The morphology evaluation of pancreatic tumor was done by a senior radiology physician.The peripancreatic vascular invasion was evaluated using a TVC computed tomography grading system by two senior radiology physicians.Two physicians read collectively films and then achieved consistent results if there was a disputed result.Observation indicators:(1) tumor detection and morphology evaluation by MDCT;(2) detection of processing images on vascular invasion signs and vascular anatomical variations after MDCT;(3) TVC grading results of peripancreatic vascular invasion by MDCT;(4) correlation between TVC grading results of peripancreatic vascular invasion by MDCT and surgical grading results;(5) follow-up and survival situations.The follow-up using outpatient examination and telephone interview was per-formed to detect postoperative survival up February 2018.Measurement data with normal distribution were represented as x±s.Measurement data with skewed distribution were described as M (range).The measured values of tumor diameter by MDCT and surgical resection were done by Pearson correlation analysis.The Spearman analysis was used to analyze the correlation in the consistency of the vascular invasion results evaluated by two viewers and between TVC grading evaluation results and surgical grading results.Results (1) Tumor detection and morphology evaluation by MDCT:66 patients underwent MDCT,nCTCP combined with MPR images showed that there were 63 low-density tumors and 3 isopycnic tumors.The nCTCP images of 66 patients showed that 42 had biliary system dilatation and pancreatic duct dilatation,11 had pancreatic duct dilatation,5 had pancreatic duct break and 8 didn't have obvious signs of biliary and pancreatic duct dilatation and break.Results of MDCT of 66 patients showed that 48 tumors were located in head or uncinate process of pancreas,18 in the body and tail of pancreas,showing a consistency with surgical results.Pearson correlation analysis showed that measured values of tumor diameter was (2.7± 1.4) cm (range,1.3-7.2 cm) in 50 patients with radical resection,with a correlation with measured values of tumor diameter of surgical resection (r =0.904,P<0.05).(2) Detection of processing images on vascular invasion signs and vascular anatomical variations after MDCT:results of MDCT in 2 patients with radical resection showed that vascular variations were confirmed intraoperatively;left gastric artery directly started with celiac axis in 1 patient,common hepatic artery started with superior mesenteric artery,and celiac axis was invaded by tumor tissues,with grading 2 of TVC grading evaluation;accessory left hepatic artery in 1 patient started with gastroduodenal artery and was invaded by tumor tissues,with grading 2 of TVC grading evaluation.Results of MDCT in 2 patients with palliative operation showed peripancreatic vascular invasion,tumor of 1 patient invaded inferior vena cava,and tumor of other patient invaded superior mesenteric artery and involved the first branch of superior mesenteric artery.(3) TVC grading results of peripancreatic vascular invasion by MDCT:there was an overall positive correlation of TVC grading evaluation results in 317 peripancreatic vessels invasion of 66 patients between two physicians (r =0.827,P<0.05).There was a positive correlation of TVC grading evaluation results in celiac axis,common hepatic artery,superior mesenteric artery,superior mesenteric vein and portal vein between two physicians (r=0.661,0.911,0.809,0.911,0.614,P<0.05).(4) Correlation between TVC grading results of peripancreatic vascular invasion by MDCT and surgical grading results:there was an overall positive correlation in 317 peripancreatic vessels invasion of 66 patients between TVC grading evaluation results and surgical grading results (r=0.806,P<0.05).There were positive correlations between TVC grading evaluation results and surgical grading results in the artery group (celiac axis,common hepatic artery and superior mesenteric artery) and vein group (superior mesenteric vein and portal vein) (r=0.703,0.823,P< 0.05).There were positive correlations between TVC grading evaluation results and surgical grading results in the celiac axis,common hepatic artery,superior mesenteric artery,superior mesenteric vein and portal vein (r =0.792,0.464,0.823,0.809,0.812,P<0.05).(5) Follow-up and survival situations:56 of 66 patients were followed up for 3-18 months,with a median time of 7 months.During the follow-up,53 patients had tumor metastases,including 50 deaths and 3 survivors with tumor;3 patients had tumor-free survival.Conclusion Multiple imaging techniques in the MDCT combined with assessment of vascular invasion using a TVC computed tomography grading system can accurately evaluate morphology of pancreatic tumor and peripancreatic vascular invasion,and increase accuracy of preoperative assessment of pancreatic cancer.
9.Effect of liraglutide on glucagon secretion in obese type 2 diabetic patients
Xiaofang SUN ; Yue WANG ; Wenjuan ZHAO ; Luan WANG ; Dongqing BAO ; Gengru QU ; Minxiu YAO ; Jian LUAN ; Yangang WANG ; Shengli YAN
Chinese Journal of Internal Medicine 2019;58(1):33-38
Objective To investigate the effect of liraglutide on glucagon release in obese type 2 diabetes (T2DM). Methods A multi-center, prospective, and self-comparison study was conducted in four hospitals in Qingdao. Twenty-four patients with T2DM were selected and treated with liraglutide for 12 weeks. Glucagon levels before and after treatment were detected before and 30 min, 60 min and 120 min after meals. Results After 12 weeks of treatment, the overall level of glucagon decreased, in which the differences in glucagon levels at 30 min [(220±79) ng/L vs. (203±77) ng/L, P<0.05] and 60 min [(248±119) ng/L vs. (203±82)ng/L, P<0.05] reached significance, respectively, comparing to those before treatment. The area under the curve of glucagon after treatment was significantly lower than that before treatment (438±190 vs. 389 ± 153, P<0.05). In contrast, after treatment, the overall level of C-peptide increased, especially the levels at 30 min [(1.53±1.02) nmol/L vs.(2.03±1.29) nmol/L ], 60 min [(1.93±1.19) nmol/L vs. (2.48±1.75) nmol/L] and 120 min [(2.36±1.47) nmol/L vs. (2.96±1.84) nmol/L], all P<0.05. The area under C-peptide curve increased significantly (3.6±2.2 vs. 4.6±2.9, P<0.05). Fasting plasma glucose, postprandial 2 h plasma glucose and glycosylated hemoglobin A1c were all lower than before, and the differences were statistically significant (P<0.05). Waist circumference and body mass index were significantly lower than before (P<0.05). The amount of insulin used for the treatment decreased by approximately 55.1% compared with that before liraglutide, and the difference was statistically significant (P<0.05). Conclusions Liraglutide inhibits glucagon secretion and lowers blood glucose. It can also reduce body weight, improve islet cell function and reduce insulin use in T2DM.
10.Comparison of different methods for measuring the phenylalanine concentration of phenylketonuria patients in dried blood spots
Xia ZHAN ; Wenjuan QIU ; Peicheng WANG ; Feng XU ; Peizhong BAO ; Xiaolan GAO ; Jun YE ; Xuefan GU
Chinese Journal of Laboratory Medicine 2018;41(5):361-365
Objective To compare the phenylalanine(Phe)concentration in the sample of dried blood spot,measured by four different methods:fluorescence assay,tandem mass spectrometry(MS/MS), including MS/MS Derivatized,MS/MS Non-Derivatized and MS/MS-Standard Curve.Methods A total of 204 dried blood spot(DBS)samples of phenylketonuria(PKU)patients from Shanghai Xinhua Hospital were collected in this study.Phe concentration in DBS was measured by fluorescence assay and MS /MS assay,including MS/MS Derivatized, MS/MS Non-Derivatized and MS/MS-Standard Curve.The samples were divided into low, middle and high concentration groups according to Phe concentration, which were under the 360 μmol/L,between 360 and 600 μmol/L,and over 600 μmol/L respectively.The differences among the groups were analyzed by consistency check and non-parametric test.Results The within-day and between-day precisions of MS/MS-Standard Curve assay were 4.0%-7.2%,the recoveries were 93.2%-97.3%.Consistency check showed that less than 8.1%of Phe value were out of the 95%limit of agreement among these four methods.In the low and middle concentration groups,the quartile of Phe value measured by fluorescence assay,MS/MS Standard Curve and MS/MS Derivatized were 176(118,251)μmol/L,174 (94,273)μmol/L,153(94,242)μmol/L; and 540(478,578)μmol/L,485(414,529)μmol/L,466(402,513)μmol/L,respectively.Phe value measured by fluorescence assay was significantly higher than that by MS/MS Standard Curve,the difference was 4.8%-9.0%,(Z=-3.787 to -2.674,P<0.01). Phe value obtained from MS/MS Non-Derivatized was less than that by MS/MS-Standard Curve, the difference was 3.9%-5.2%,(Z=-7.474 to -5.747,P<0.01).In the high concentration group,the quartile of Phe value measured by MS/MS-Standard Curve, MS/MS Derivatized, MS/MS Non-Derivatized and fluorescence assay were 807(695,924)μmol/L,700(575,785)μmol/L,680(623,771)μmol/L and 711(674, 794)μmol/L, respectively.Phe value obtained from MS/MS Standard Curve was significantly higher than those from the other methods and the difference was 10.9% -16.2%,(Z=-4.458 to-4.356,P<0.01).Conclusions The MS/MS Standard Curve assay showed good specificity and accuracy.These four assays displayed good agreement.Although there was difference in measuring the Phe concentration among these methods, they could be used in blood Phe concentration monitoring for PKU patients.