1.The application of certified reference materials for clinical mass spectrometry
Lingxiao SHEN ; Jinfei MA ; Qingqing YANG ; Pengyun LIU ; Huafen LIU
Chinese Journal of Laboratory Medicine 2021;44(5):437-441
Reference materials are one of the major approaches to achieve measurement accuracy and metrological comparability. Different functions of reference materials should also be distinguished when applied to mass spectrometry as an emerging technology in clinical laboratory. Proper reference materials for validation, calibration and quality control of measurement method can ensure the accuracy and comparability of test results. Based on the problems of reference materials in clinical mass spectrometry, the precautions for the use of reference materials are summarized in the aspects of measurement method validation, calibrator usage and quality control.
2.Syncope unit improves diagnosis and prognosis of patients with suspected syncope
Xuelei BAI ; Xiaodong WANG ; Derui LIU ; Zhaodi JING ; Yanjia FAN ; Mengli FAN ; Pengyun FAN ; Zizhong XI
Chinese Journal of General Practitioners 2021;20(6):662-667
Objective:To evaluate the effectiveness of syncope unit in improving the diagnosis efficiency and treatment prognosis of patients with suspected syncope.Methods:The standardized syncope unit was established in the Affiliated Nanyang Second General Hospital of Xinxiang Medical College in 2018. Patients with suspected syncope attending from November 2018 to April 2019 (before the establishment of syncope unit) and from May to October 2019 (after the establishment of syncope unit) were enrolled in the study. There were 109 cases attending before the establishment of syncope unit (control group) and 126 cases attending after establishment (syncope unit group). The positive rate of examination, the treatment and its cost before and after the establishment of syncope unit were compared. After one year, the follow-up rate, recurrence rate, rehospitalization rate, treatment satisfaction and quality of life of patients were documented and compared between two periods.Results:The positive rates of tilt table test [61.90%), Holter monitoring [64.29%(81/126)], exercise stress test [7.14%(9/126)] invasive electrophysiology [40.48%(51/126)], cardiac imaging [9.52%(12/126)] and 24-h blood pressure monitoring [55.56%(70/126)] in syncope unit group were significantly higher than those in control group [44.95%(49/109), 36.70%(40/109), 5.50%(6/109), 10.09%(11/109), 2.75%(3/109) and 40.37%(44/109); χ2=19.28, 23.11, 6.93, 28.18, 15.85 and 11.61,respectively; all P<0.01]. The diagnostic rate of etiology in syncope unit group was significantly higher than that in control group [87.30%(110/126) vs. 77.06%(84/109), χ2=21.70, P<0.01].The time from onset to cardiac assessment and hospitalization time in syncope unit group were significantly shorter than those in control group[(3.68±1.93)h vs. (7.31±2.64)h;(6.17±1.52)d vs. (10.83±2.09)d]. The hospitalization rate [3.17%(4/126) vs. 8.26%(8/109)], hospital mortality [0.79%(1/126) vs. 2.75%(3/109)] and treatment cost [(4.91±1.14) thousands Yuan vs. (7.05±2.53) thousands Yuan] in syncope unit group were significantly lower than those in control group ( t=14.49, P<0.01; t=8.62, P=0.02;χ2=15.83, P<0.01;χ2=10.03, P=0.01; t=6.17, P=0.03).The outpatient follow-up rate [82.54%(104/126)] and treatment satisfaction rate [91.35%(95/104)] in syncope unit group were significantly higher than those in control group [61.47%(67/109) and 64.18%(43/67)]; and the recurrence rate [14.42%(15/104)] and rehospitalization rate [7.69%(8/104)] in syncope unit group were significantly lower than those in control group [40.30%(27/67) and 23.88%(16/67)](χ2=17.30, 20.37, 18.56, 15.08,all P<0.01). The scores of psychological status, physiological status, environmental status, social relations and overall quality of life in contral group were significantly lower than those in syncope unit group (43.62±12.84 vs. 59.13±13.95,43.10±11.31 vs. 5.86±12.09,52.35±12.76 vs. 63.58±13.05,54.87±12.08 vs. 67.91±14.23,58.42±11.87 vs. 69.28±13.51; t=7.74, P=0.03; t=7.50, P=0.03; t=8.66, P=0.02; t=9.77, P=0.01; t=8.46, P=0.02, respectively). Conclusion:The establishment of standardized syncope unit is helpful to improve the diagnosis efficiency and the prognosis of patients with suspected syncope, and also reduce the cost of diagnosis and treatment.
3.Preparation and measurement of sodium tanshinone Ⅱ-A sulfonate in mice serum and effects on big conductance calcium-activated potassium channels in porcine coronary artery smooth muscle cells
Jun CHENG ; Xiaorong ZENG ; Pengyun LI ; Miaoling LI ; Zhifei LIU ; Jie PEI ; Yan YANG
Chongqing Medicine 2015;(4):436-438
Objective To discuss a method combining serum‐pharmacology and electrophysiology technology ,and to research the mechanism of dilating porcine coronary artery of sodium tanshinone Ⅱ‐A sulfonate (DS‐201) .Methods To give mice intragastric administration solution and measure DS‐201 concentration in mice serum ,and apply the serum to single channel patch to research its effect on big conductance calcium‐activated potassium channels(BKca ) in porcine coronary artery smooth muscle cells (PCASMCs) . Results The linear range of concentration containing DS‐201 serum was 0 .73 to 1 .91 μg/mL (r=0 .997 7) ,the recycle rate was 99 .85% -101 .47% ,and the concentration was(7 .32 ± 4 .25)μg/mL ;the result indicates that serum containing DS‐201 has activa‐tion effects on BKCa in PCASMCs ,while there was no statistical significance (P>0 .05) .Conclusion The establishment method of the alcohol extraction of Danshen is useful and reliable .The HPLC method of measuring DS‐201 concentration is precise .Choosing higher quality drugs or raising bioavailability can improve combination of the serum pharmacology and electrophysiological tech‐nique .
4.Efficacy of S-1 combined with cisplatin as first-line chemotherapy for advanced AFP positive gastric cancer
Wanlu LI ; Feng WANG ; Bo WANG ; Pengyun NIU ; Qingcun LIU ; Qingxia FAN
Chinese Journal of Clinical Oncology 2016;43(4):152-155
Objective:To comparatively analyze the efficacy of S-1 plus cisplatin as first-line chemotherapy between advanced AFP posi-tive and AFP negative gastric cancer. Methods:A total of 89 eligible patients with advanced gastric cancer from January 2010 to June 2013 were enrolled in this retrospective study. The cases were divided into AFP positive group (n=18) and AFP negative group (n=71) based on serum AFP level before treatment. Both groups received S-1 plus cisplatin as first-line treatment. Results:Significant differ-ences were observed in remission rate (66.7% versus 32.4%, P=0.028) and radical surgical resection rate (44.5% versus 18.3%, P=0.029) after chemotherapy between AFP positive group and AFP negative group. In the two groups, neither progression-free survival nor overall survival (OS) showed any significance (P>0.05) in patients without surgery after chemotherapy. However, the disease-free survival of the two groups with radical surgical resection after chemotherapy was significantly different (median:27.0 months versus 15.6 months, P=0.034). The OS of the AFP positive group was evidently longer than that of the AFP negative group (median:16.0 months versus 11.0 months, P=0.005). Conclusion:As first-line chemotherapy, S-1 plus cisplatin was more effective for the advanced AFP positive gastric cancer and prolonged overall survival.
5.Analysis of the incidence of syncope and the influencing factors of death in patients with cardiovascular critical emergency
Xuelei BAI ; Xiaodong WANG ; Yingli ZHANG ; Derui LIU ; Zhaodi JING ; Mengli FAN ; Yanjia FAN ; Pengyun FAN
Chinese Critical Care Medicine 2021;33(3):324-328
Objective:To observe the incidence of syncope in patients with acute and critical cardiovascular diseases and to explore the risk factors of death.Methods:925 cases of acute heart failure, acute myocardial infarction, pulmonary embolism, arrhythmia and aortic dissection rupture who participated in Prospective, Multi-CenterRegistered Research Project for Chinese Syncope Patients from March 2018 to March 2020, admitted to the department of emergency of Nanyang Second General Hospital were selected as the research objects. The incidence and mortality of syncope were recorded, and the patients were divided into syncope group and non-syncope group according to whether they were accompanied by syncope or not. The incidence of syncope in male and female patients with different cardiovascular critical diseases, the age and mortality of cardiovascular critical patients with syncope or not were analyzed and compared. Multivariate Logistic regression analysis was used to analyze the risk factors of death, and receiver operating characteristic curve (ROC curve) was drawn to evaluate the predictive value of risk factors on the prognosis of patients.Results:The incidence of syncope in 5 kinds of cardiovascular critical patients from high to low was: acute myocardial infarction 3.03% (28/925), arrhythmia 2.70% (25/925), pulmonary embolism 1.51% (14/925), aortic dissection rupture 1.41% (13/925), acute heart failure 0.65% (6/925), with statistically significant differences ( χ2 = 10.765, P = 0.010). There was no significant difference in the incidence of syncope between male and female patients with pulmonary embolism, aortic dissection rupture, acute myocardial infarction, arrhythmia and acute heart failure. The age of patients with aortic dissection rupture, acute myocardial infarction and arrhythmia in syncope group were significantly higher than those in non-syncope group [aortic dissection rupture (years old): 66.29±15.64 vs. 57.63±14.23, acute myocardial infarction (years old): 69.55±15.13 vs. 62.10±15.75, arrhythmia (years old): 70.48±14.93 vs. 60.29±16.31, all P < 0.05]. The mortality of patients with pulmonary embolism, aortic dissection rupture, acute myocardial infarction, arrhythmia, acute heart failure in syncope group were significantly higher than those in non-syncope group [pulmonary embolism: 5.81% (5/86) vs. 0.95% (8/839), aortic dissection rupture: 4.65% (4/86) vs. 0.60% (5/839), acute myocardial infarction: 4.65% (4/86) vs. 1.19% (10/839), arrhythmia: 2.33% (2/86) vs. 0.95% (8/839), acute heart failure: 2.33% (2/86) vs. 0.60% (5/839), all P < 0.05]. Multivariate Logistic regression analysis showed that age [odds ratio ( OR) = 2.158, 95% confidence interval (95% CI) was 0.921-4.785, P = 0.000], pulmonary embolism ( OR = 15.391, 95% CI was 8.904-27.314, P = 0.001), aortic dissection rupture ( OR = 13.079, 95% CI was 6.237-25.509, P = 0.000), acute myocardial infarction ( OR = 18.826, 95% CI was 10.420-32.921, P = 0.000), syncope ( OR = 4.940, 95% CI was 1.764-9.287, P = 0.000) were risk factors for the prognosis of patients with acute and critical cardiovascular diseases. ROC curve analysis showed that syncope had a certain predictive value for 28-day prognosis of patients [the area under the ROC curve (AUC) = 0.760, P = 0.000], when the cut-off value was 4.12, the sensitivity was 88.51%, the specificity was 78.05%, the positive predictive value was 81.31%, and the negative predictive value was 84.27%. Conclusions:Syncope is an independent risk factor of death in patients with acute and critical cardiovascular diseases. For patients with syncope as the chief complaint, we should quickly identify the types of acute and critical diseases and assess the risk of sudden death.
6.Correlation between long non-coding RNA-LOC391533 and inadequate placental spiral artery remodeling in patients with severe preeclampsia
Shihong CUI ; Jie YU ; Ling LIU ; Juan CHEN ; Fengjuan SHI ; Ting ZHANG ; Pengyun LI
Chinese Journal of Perinatal Medicine 2017;20(10):718-723
Objective To investigate the correlation between long non-coding RNA (lncRNA)-LOC391533 and inadequate placental spiral artery remodeling in severe preeclampsia (sPE).Methods Thirty-six gravidas who were admitted to the Third Affiliated Hospital of Zhengzhou University with sPE from January 2016 to December 2016 were enrolled in sPE group.An equal number of healthy gravidas who experienced uneventful pregnancy and were of similar age (difference less than two years) and gestational age (difference less than one week) to those in the sPE group served as controls.Scanning electron microscopy was used to measure the luminal area and vessel wall thickness of placental spiral arteries for all gravidas.Levels of vascular endothelial growth factor (VEGF) and soluble VEGF receptor-1 (sVEGFR-1) in placenta tissues and maternal serum samples were detected by Western blot and ELISA.Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of LOC391533 at mRNA level in placenta tissues of the two groups.Independent two samples t-test,Mann-Whitney U test and Spearman correlation analysis were used for statistical analysis.Results (1) The average luminal area of spiral arteries of the sPE group was smaller than that of the control group [(130.1 22.3) vs (188.1 ±21.5) μ m2,t=10.888,P<0.05],but the average thickness of spiral artery wall was thicker [(122.619.5) vs (98.9±2.5) μ m,t=-8.812,P<0.05].(2) Compared with the control group,the sPE group showed increased sVEGFR-1 at protein level in both placenta tissues and serum samples [placenta:0.2±0.0 vs 0.4±0.1,serum:(15.6±2.4) vs (50.8±6.1) ng/L,t=-17.569 and-30.699,both P<0.05],decreased VEGF at protein level in both placenta tissues and serum samples [placenta:0.6 ± 0.1 vs 0.2±0.0,serum:(40.8±3.2) vs (28.1 ±3.2) ng/L,t=18.013 and 16.200,both P<0.05],and enhanced expression of LOC391533 at mRNA level in placenta tissues (1.00.2 vs 2.40.5,t=-14.799,P<0.05).(3) Expression of LOC391533 at mRNA level in placenta tissues of the sPE group was positively correlated with spiral artery wall thickness and levels of sVEGFR-1 protein in placenta tissues and serum (r=0.683,0.759 and 0.857,all P<0.05),and negatively correlated with luminal area and levels of VEGF protein in placenta tissues and serum (r=-0.702,-0.806 and-0.796,all P<0.05).Conclusions Abnormal expression of VEGF and sVEGF-1 in placenta and serum of patients with sPE may be related to inadequate placental spiral artery remodeling.
7.Application of LC-MS/MS in the accurate assessment of vitamins
Xiaofen YUAN ; Jinfei MA ; Qingqing YANG ; Pengyun LIU ; Lingxiao SHEN ; Qianjun ZHANG ; Yikun LI ; Huafen LIU
Chinese Journal of Laboratory Medicine 2019;42(6):479-483
Vitamins are classified as either fat-soluble (vitamins A, D, E, K) or water-soluble (vitamins B and vitamin C). Traditional methods of immunoassay have only been developed for vitamins D,B6, B9 and B12. However, they cannot distinguish between vitamin subtypes such as D2, D3 and associated epi isomers (which has higher leveks in infants),giving false positive or negative results. Mass spectrometry has become a gold standard method for small molecule analysis in biological samples with its advantages in speed,resolution,sensitivity and specificity. It is widely used in clinical research and diagnosis and provides an efficient method for simultaneous detection of multivitamins in one injection using one low volume sample collection.
8.Genetic diagnosis and analysis of eight cases with central 22q11.2 deletion syndrome
Jing GUO ; Pengyun LI ; Jia CHE ; Shanshan ZHAI ; Weifang TIAN ; Ying LI ; Hua ZHANG ; Ling LIU
Chinese Journal of Medical Genetics 2024;41(2):145-149
Objective:To explore the pregnancy outcome and postpartum clinical phenotype of LCR22B/C~D central 22q11.2 deletion syndrome.Methods:For fetuses diagnosed with central 22q11.2 deletion by chromosomal microarray analysis (CMA) at the Prenatal Diagnosis Center of the Third Affiliated Hospital of Zhengzhou University from January 2019 to April 2022, their prenatal imaging, parental CMA verification, pregnancy outcomes and postpartum clinical phenotype were analyzed.Results:Eight cases of central 22q11.2 deletion syndrome were included, including six cases with LCR22B~D 22q11.2 deletions and two with LCR22C~D 22q11.2 deletions. Among the six cases with LCR22B~D type 22q11.2 deletions, three had shown cardiovascular malformations (right aortic arch, ventricular septal defect, mild tricuspid regurgitation), one had shown urinary defect (right kidney heterotopia). Two cases with LCR22C~D 22q11.2 deletions showed nonspecific ultrasonographic findings, including oligohydramnios with growth restriction and nuchal skin thickening. The CMA verification showed that six cases were inherited from their parents, and five couples had chosen to continue with the pregnancy. Postpartum follow-up showed that the physical and intellectual development of all children were normal. One couple had opted to terminate the pregnancy considering the ectopic fetal right kidney. Two remaining cases had decided to terminate their pregnancies without parental verification.Conclusion:The central 22q11.2 deletion syndrome of the LCR22B/C~D type is different from the classical types. Its genetic information mainly comes from parents. Prenatal imaging has mainly shown cardiovascular and urinary abnormalities. Postnatal growth and intellectual development have been normal. Therefore, the couples should be provided with suffice prenatal genetic counseling.
9.Molecular mechanism of miR-369-3p regulating hepatocellular carcinoma cell proliferation and apoptosis by targeting ACTN4
Hongfeng ZHAO ; Meng LOU ; Minping BI ; Xiaowei YANG ; Pengyun HU ; Jialin LIU
Chinese Journal of Oncology 2021;43(11):1156-1163
Objective:To investigate the effect of miR-369-3p targeting ACTN4 expression on proliferation and apoptosis of hepatocellular carcinoma cells.Methods:Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were used to detect the expression levels of miR-369-3p and ACTN4 in hepatocarcinoma tissues and adjacent tissues. MiR-369-3p mimics, miR-negative control (NC), si-ACTN4, and si-NC were transfected into hepatocellular carcinoma MHCC97H cells by liposome method. Cell proliferation was detected by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-dipheny-ltetrazolium bromide (MTT) assay. Flow cytometry was used to detect cell cycle and apoptotic rates. The dual luciferase reporter assay was used to verify the targeted regulation of ACTN4 by miR-369-3p. Western blot was used to detect the expressions of cyclin D1, p21, Bcl-2 and Bax.Results:The expression level of miR-369-3p in liver cancer tissue was lower than that in adjacent tissues [(0.46±0.04) vs (1.00±0.08), P<0.001)], while the expression level of ACTN4 was higher than that in adjacent tissues [mRNA (3.12±0.29) vs (1.01±0.09); protein (0.61±0.06) vs (0.25±0.03), P<0.001]. Overexpression of miR-369-3p significantly decreased the cell viability[(0.71±0.06) vs (1.26±0.11), P<0.001)], increased cell apoptosis rate [(20.16±2.11)% vs (6.25±0.64)%, P<0.001], increased the proportion of cells in G 1 phase [(31.14±3.36)% vs (51.56±5.23)%, P<0.001], decreased the proportion of cells in S phase [(32.44±3.56)% vs (14.33) ±1.45)%, P<0.001], increased the levels of p21 and Bax protein ( P<0.001), and decreased the levels of cyclin D1 and Bcl-2 protein ( P<0.001). Inhibition of the expression of ACTN4 significantly reduced the cell viability [(0.78±0.07) vs (1.24±0.12), P<0.001], increased the apoptosis rate [(6.58±0.66)% vs (18.32±1.82)%, P<0.001], increased the proportion of cells in G 1 phase [(48.69±4.21)% vs (30.33±3.01)%, P<0.001], decreased the proportion of cells in S phase [(36.21±3.42)% vs (18.54±1.61)%, P<0.001], increased the protein levels of p21 and Bax ( P<0.001), and decreased the levels of cyclin D1 and Bcl-2 protein ( P<0.001). Compared with the miR-369-3p+ pcDNA group, overexpression of ACTN4 increased the proliferation ability of hepatocellular carcinoma MHCC97H cells at 72 hours of culture[(1.12±0.11) vs (0.68±0.06), P<0.001], significantly reduced the proportion of cells in G 1 stage [(38.81±3.24)% vs (51.80±4.57)%, P<0.001], significantly increased the proportion of S-phase cells [(31.65±3.11)% vs (15.69±1.44)%, P<0.001], decreased cell apoptosis rate [(13.86±1.37)% vs (22.69±2.24)%, P<0.001], increased protein expressions of cyclin D1 and Bcl-2 ( P<0.001), decreased the protein expressions of p21 and Bax ( P<0.001). Conclusion:MiR-369-3p can induce cell cycle arrest in G 1 phase, inhibit the proliferation and promote apoptosis of liver cancer cells by regulating the expression of ACTN4.
10.Molecular mechanism of miR-369-3p regulating hepatocellular carcinoma cell proliferation and apoptosis by targeting ACTN4
Hongfeng ZHAO ; Meng LOU ; Minping BI ; Xiaowei YANG ; Pengyun HU ; Jialin LIU
Chinese Journal of Oncology 2021;43(11):1156-1163
Objective:To investigate the effect of miR-369-3p targeting ACTN4 expression on proliferation and apoptosis of hepatocellular carcinoma cells.Methods:Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were used to detect the expression levels of miR-369-3p and ACTN4 in hepatocarcinoma tissues and adjacent tissues. MiR-369-3p mimics, miR-negative control (NC), si-ACTN4, and si-NC were transfected into hepatocellular carcinoma MHCC97H cells by liposome method. Cell proliferation was detected by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-dipheny-ltetrazolium bromide (MTT) assay. Flow cytometry was used to detect cell cycle and apoptotic rates. The dual luciferase reporter assay was used to verify the targeted regulation of ACTN4 by miR-369-3p. Western blot was used to detect the expressions of cyclin D1, p21, Bcl-2 and Bax.Results:The expression level of miR-369-3p in liver cancer tissue was lower than that in adjacent tissues [(0.46±0.04) vs (1.00±0.08), P<0.001)], while the expression level of ACTN4 was higher than that in adjacent tissues [mRNA (3.12±0.29) vs (1.01±0.09); protein (0.61±0.06) vs (0.25±0.03), P<0.001]. Overexpression of miR-369-3p significantly decreased the cell viability[(0.71±0.06) vs (1.26±0.11), P<0.001)], increased cell apoptosis rate [(20.16±2.11)% vs (6.25±0.64)%, P<0.001], increased the proportion of cells in G 1 phase [(31.14±3.36)% vs (51.56±5.23)%, P<0.001], decreased the proportion of cells in S phase [(32.44±3.56)% vs (14.33) ±1.45)%, P<0.001], increased the levels of p21 and Bax protein ( P<0.001), and decreased the levels of cyclin D1 and Bcl-2 protein ( P<0.001). Inhibition of the expression of ACTN4 significantly reduced the cell viability [(0.78±0.07) vs (1.24±0.12), P<0.001], increased the apoptosis rate [(6.58±0.66)% vs (18.32±1.82)%, P<0.001], increased the proportion of cells in G 1 phase [(48.69±4.21)% vs (30.33±3.01)%, P<0.001], decreased the proportion of cells in S phase [(36.21±3.42)% vs (18.54±1.61)%, P<0.001], increased the protein levels of p21 and Bax ( P<0.001), and decreased the levels of cyclin D1 and Bcl-2 protein ( P<0.001). Compared with the miR-369-3p+ pcDNA group, overexpression of ACTN4 increased the proliferation ability of hepatocellular carcinoma MHCC97H cells at 72 hours of culture[(1.12±0.11) vs (0.68±0.06), P<0.001], significantly reduced the proportion of cells in G 1 stage [(38.81±3.24)% vs (51.80±4.57)%, P<0.001], significantly increased the proportion of S-phase cells [(31.65±3.11)% vs (15.69±1.44)%, P<0.001], decreased cell apoptosis rate [(13.86±1.37)% vs (22.69±2.24)%, P<0.001], increased protein expressions of cyclin D1 and Bcl-2 ( P<0.001), decreased the protein expressions of p21 and Bax ( P<0.001). Conclusion:MiR-369-3p can induce cell cycle arrest in G 1 phase, inhibit the proliferation and promote apoptosis of liver cancer cells by regulating the expression of ACTN4.