1.An investigation on the accuracy of a simple measurement method of glomerular filtration rate
Ke GONG ; Yingshu ZOU ; Haifeng ZHU ; Xu PENG ; Dongfang QIN ; Bixuan SUN ; Xuejing WANG
Chinese Journal of Laboratory Medicine 2022;45(7):738-744
Objective:To evaluate the proper blood collection time and calculation formula by measuring the iohexol plasma clearance as a representative of glomerular filtration rate at the same time of routine enhanced computed tomography (CT) examination.Methods:The prospective study method was applied, and 9 subjects with normal renal function, who admitted in Civil Aviation General Hospital from September 2018 to June 2019, were included. A single bolus of a standard dose (5 ml) (iodine concentration: 350 mgI/ml) was injected. The concentration of iohexol was measured from heparin plasma at fasting state of the subject and at nine different times after the injection, respectively. More than 24 hours after the injection of the standard dose, an enhanced CT-level dose (50 ml) of iohexol was injected to the subject and the concentration of iohexol was measured at similar time points as the standard dose. Using a multi-point method of a standard dose as the standard, the clearance rate was calculated by three kinds of formulas including Groth and Aasted formula, Jacobsson formula and Fleming formula with the single-point method to assess iohexol plasma clearance at 0.5 to 8.0 hours post injection of enhanced CT-level dose. The correlation consistency and accuracy of the multi-point method and the single-point method, as well as the dual-point method and the single-point method were compared, and the proper blood collection time and calculation formula of the single-point method at regular enhanced CT-level dose were evaluated. The correlation between the multi-point method and the single-point method, as well as the dual-point method and the single-point method were assessed using Pearson correlation coefficient; the consistency between the multi-point method and the single-point method, as well as the dual-point method and the single-point method were assessed by bias using mean±standard deviation ( SD) and 95% confidence interval ( CI) of mean difference and so on. We assessed the concordance of GFR using GFR±5% ( P5),±10% ( P10) and 1±30% ( P30) intervals. Results:Compared with the multi-point method, the mean deviation of iohexol plasma clearance obtained by the three single-point methods increased gradually from 5 hours after the injection of iohexol ( P<0.05). Compared with the multi-point method, only 3 h results, which was calculated by the Groth and Aasted formula, reached a P value greater than 0.05, a correlation coefficient of 0.938, a mean deviation of (-5.2±8.8) ml·min -1·1.73 m -2, and the concordances were 100% corresponding to P30,77.8% corresponding to P10, and 66.7% corresponding to P5; the 2, 3 and 4 hours results, which was calculated by the Jacobsson formula, reached P values greater than 0.05, when the blood collection time was 3 hours, the correlation coefficient was 0.938, and the mean deviation was the smallest, which was (1.5±6.2) ml·min -1·1.73 m -2, and the concordances were 100% corresponding to P30, 88.9% corresponding to P10, and 66.7% corresponding to P5; the 2 and 3 hours results, which was calculated by the Fleming formula, reached P values greater than 0.05, when the blood collection time was 2 h, the correlation coefficient was 0.956, and the mean deviation was the smallest, which was (-4.5±8.8) ml·min -1·1.73 m -2, and the concordances were 100% corresponding to P30, 77.8% corresponding to P10, and 55.6% corresponding to P5,Compared with the dual-point method, when Groth or Aasted formula was used, the mean deviation was the smallest at 3 hours, which was (-5.3±5.7) ml·min -1·1.73 m -2; when Jacobsson formula was used, the mean deviation was the smallest at 2 hours, which was (1.6±1.6) ml·min -1·1.73 m -2; when Fleming formula was used, and the mean deviation was the smallest at 2 hours, which was (-4.6±4.0) ml·min -1·1.73 m -2. Conclusion:At a regular enhanced CT-level dose, one blood collection can accurately measure the glomerular filtration rate, the proper time for blood collection can be 3 hours after iohexol injection, and the appropriate calculation formula can be Jacobsson formula.
2.Qualititative analysis of 5-methyltertrahydrofolate in human serum by ID-LC-MS/MS
Xu PENG ; Yingshu ZOU ; Zongbing YANG ; Jun WANG
Chinese Journal of Clinical Laboratory Science 2018;36(3):161-165
Objective To develop an isotope dilution liquid chromatography tandem mass spectrometry (ID-LC-MS/MS) method for determining the concentration of 5-methyltertrahydrofolate (5MT) in human serum.Methods After qualitative analysis of 5MT reference,its purity was identified by two methods with different principle,i.e.,mass balance and quantitative nuclear magnetic resonance.The definite value and uncertainty of 5MT concentrations in 6 serum samples containing same level of 5MT were assessed by ultra performance liquid chromatography-triple quadrupole tandem mass spectrometer equipped with electrospray ionization in monitoring mode of multiple reactions,in which 13C5-5MT was used as the internal standard.The precision,accuracy,limit of detection (LOD) and limit of quantification (LOQ) of the method were also evaluated.Results After qualitative analysis for confirming the principal component in the sample,the purity of 5MT reference was measured as 76.65% by mass balance and quantitative nuclear magnetic resonance method.The definite value of 5MT in the serum sample was 8.22 ng/mL,the extended uncertainty was 0.54 ng/mL,LOD was 0.05 ng/mL,LOQ was 0.50 ng/mL and the repeatability and inter-batch precision CV were within 5.0%.The accuracy of measured results was verified by using NIST SRM 1955 since all the values of 3 levels were within the range of certified value with the extended uncertainty.Conclusion A accurately quantitative determination for the concentration of 5 MT in human serum with high specificity and sensitivity was established by using ID-LC-MS/MS method.
3.A review of effects of disrupted primary caregiver infant relationship on brain development and psychopathology
Zhenyu SHI ; Wenqing ZHAO ; Danping WU ; Fenghuan LUAN ; Hui LIU ; Juan DONG ; Yingshu ZHANG ; Han ZOU ; Wen XU ; Xudong ZHAO
Chinese Mental Health Journal 2018;32(2):130-135
The disrupted caregiver infant relationships are not unpopular (infant abuse and non-traumatic caregiver-infant relationship).Animal and human studies have demonstrated this disruptive relationship exerts the fundamental and enduring impacts on stress system, limbic system and relevant cortex.Individual carrying such biological susceptibility might develop psychopathology under stresses at later life.The assessment needs to get involved the measures with more objectivity,real time and moment-to-moment components.The study on caregiverinfant relationship also should focus on the historical and cultural aspects of China.
4. Lithocarpus polystachyus (Sweet Tea) water extract promotes human hepatocytes HL7702 proliferation through activation of HGF/AKT/ERK signaling pathway
Ming LEI ; Nana CHEN ; Yingshu XU ; Jianmei GAO ; Ming LEI ; Nana CHEN ; Qihai GONG ; Jianmei GAO
Chinese Herbal Medicines 2022;14(4):576-582
Objective: Sweet Tea (ST), derived from the leaves of Lithocarpus polystachyus, is a Chinese folk medicine with wide pharmacological activities. However, the promotive effects of ST water extract on hepatocytes proliferation and its underlying mechanism remains still unknown. In the present study, the beneficial effects of ST water extract on human hepatocytes and its possible mechanism were investigated. Methods: MTT assay was used to detect the safety range of ST; HL7702 cells were divided into four groups: control group, ST low- (50 μg/mL), medium- (200 μg/mL) and high-concentration (800 μg/mL) groups; BrdU ELISA and EDU staining were used to observe DNA content and cell proliferation; Moreover, flow cytometry was applied to analyze the distribution of cell cycle. Furthermore, the expression of cyclin D1, CDK4, HGF/c-Met, Akt, Erk1/2 were detected by Western blot. Results: It was found that ST water extract concentration-dependent promoted human hepatocytes HL7702 cell proliferation within 72 h through accumulating the cells in S phase and G2/M phase. Furthermore, ST water extract up-regulated expression of Cyclin D1 and CDK4 proteins. Moreover, ST water extract not only increased HGF expression and phosphorylation of c-Met level, but also activated the phosphorylation levels of AKT, ERK1/2. Interestingly, both of AKT inhibitor A6730 and ERK1/2 inhibitor U0126 reversed the promotive effects of ST water extract, which further confirmed that activation of AKT and ERK1/2 were involved. Conclusion: The findings reveal that ST water extract promoted HL7702 cells proliferation through the stimulation of cell cycle mediated by activating the AKT- and ERK1/2-related pathway.