[Alternative Views of Transesophageal Echocardiography in the Assessment of Aorta Structure].

OBJECTIVE: To investigate the imaging views and scanning procedure of the aorta using the transesophageal echocardiography (TEE). METHODS: In accordance with the established TEE scanning process, 337 patients had their aortas scanned on four different levels, including upper, middle, lower segment of esophagus and gastric base. All parameters such as the depth of probe, imaging angle were recorded. RESULTS: A total of 330 cases of patients completed the procedure, the other 7 cases experienced the failure of TEE probe placement. Five patients received the new acoustic window for TEE imaging of large vessels anterior to the trachea by using a saline-filled endotracheal balloon, for the main structures anterior to the trachea were poorly visualized. The average time-consuming of the scanning was (15.6 ± 3.8) min. Depth of the probe, imaging angle and the aortic wall changed with the changing of imaging views. CONCLUSION: It is important to be familiar with the aorta anatomy and TEE imaging features for the purpose of overcoming the difficult in the assessment of aorta on TEE imaging.

Calcium regulates the physiological and molecular responses of Morus alba roots to cadmium stress.

Heavy metal cadmium (Cd) is toxic to organisms. Mulberry (Morus alba L.) is a fast-growing perennial that is also an economical Cd phytoremediation material with large biomass. However, the molecular mechanisms underlying its Cd tolerance remain unclear. Here, we reveal the physiological and molecular mechanisms underlying Cd toxicity under varying calcium (Ca) treatments. First, under low-Ca treatment (0.1 mM Ca), mulberry growth was severely inhibited and the root surface structure was damaged by Cd stress. Second, electrophysiological data demonstrated that 0.1 mM Ca induced an increased Cd2+ influx, leading to its accumulation in the entire root and root cell walls. Third, high-Ca treatment (10 mM Ca) largely alleviated growth inhibition, activated antioxidant enzymes, increased Ca content, decreased Cd2+ flux, and inhibited Cd uptake by roots. Finally, 0.1 mM Ca resulted in the activation of metal transporters and the disruption of Ca signaling-related gene expression, which facilitated Cd accumulation in the roots, aggravating oxidative stress. These adverse effects were reversed by treatment with 10 mM Ca. This study preliminarily revealed the mechanism by which varying Ca levels regulate Cd uptake and accumulation in mulberry roots, provided an insight into the interrelationships between Ca and Cd in the ecological and economic tree mulberry and offered a theoretical basis for Ca application in managing Cd pollution.

[Correlation between 2D and 3D echocardiography in measuring aortic annuals diameter].

OBJECTIVE: To compare the results of 2D and 3D echocardiography in measuring aortic annulus diameter (AAD). METHODS: Preoperitve transesophageal echocardiography (TEE), transthoracic echocardiography (TTE) and 3D transesophageal echocardiography (RT 3D TEE) were performed on 52 patients who underwent aortic valve replacements to measure AAD. The parameters were compared between systole and diastole. RESULTS: The parameters in systole were significant greater than those in diastole (P < 0.05). No significant differences in the AAD value were found between TTE and TEE. However, the AAD values determined with TTE and TEE were different from those determined with RT 3D TEE (P < 0.05). The two-dimensional measurements produced smaller AAD values in both systole and diastole than the 3D measurements (P < 0.001). TEE correlated well with RT 3D TEE, with the r value of 0.775 and 0.765 for systole and diastole, respectively. The regression was 3D-AADs = 0.531 x TEE-AADs + 19.879 (mm). CONCLUSION: RT 3D TEE can accurately reconstruct the 3D structure of aortic annulus and calculate the aortic annulus dimension. TEE-AAD correlates well with 3D-AAD.

[Echocardiographic features of patients with anomalous left coronary artery from the pulmonary artery].

OBJECTIVE: To summarize the echocardiographic features of anomalous left coronary artery from the pulmonary artery (ALCAPA). METHODS: A study was performed on 10 patients with ALCAPA retrospectively and the result was compared with coronary angiography to summarize its echocardiographic features. RESULTS: Of the 10 patients with ALCAPA, there were 3 cases of infantile type and 7 cases of adult type. Left ventricular cavity was enlarged and the anterior and lateral wall motion was reduced in all patients with the infantile type ALCAPA. The trunk and branch of the right coronary artery was enlarged in patients with the adult type, meanwhile the diastolic blood flow signal could be detected in inter-ventricular septum and in the area supplied by the left anterior descending coronary artery. The trunk of the left coronary artery originated from the pulmonary artery in 7 patients, the left circumflex artery originated from the pulmonary artery in another 3 patients. Diastolic blood flow signal could be detected by the Doppler echocardiogram in the pulmonary artery in all patients. One patient was misdiagnosed as coronary artery fistula, and correct diagnosis was made by the coronary angiography. In 1 patient, echocardiography examination suggested that the left coronary artery was originated from left coronary aortic sinus and angiography results showed that the left coronary artery was originated from the right posterior sinus of the pulmonary artery. Eight ALCAPA patients suffered from other cardiovascular abnormalities, including atrial septum defect, ventricular septal defect and non-compaction of ventricular myocardium. CONCLUSIONS: Abnormal connection of the left coronary artery with pulmonary artery and diastolic blood flow are typical echocardiographic findings in patients with ALCAPA. Coronary angiography serves as an important tool for the correct diagnosis of ALCAPA.

Promoter of TFPI-2 is hypermethylated in Chinese pediatric acute myeloid leukemia.

Human tissue factor pathway inhibitor-2 (TFPI-2) has been implicated as a metastasis-associated gene in many types of tumors. In this study, we investigated whether TFPI-2 was inactivated epigenetically in pediatric acute myeloid leukemia (AML). Methylation status was investigated by methylation-specific polymerase chain reaction and bisulfate genomic sequencing. TFPI-2 was aberrantly methylated in 50% (3/6) of AML cell lines. Aberrant methylation of TFPI-2 promoter was detected in 71.6% (48/67) of the Chinese pediatric AML patients. TFPI-2 transcript was significantly lower in AML group compared with controls (3.44 vs. 32.8, P<0.001). Patients with methylated TFPI-2 gene had significantly lower TFPI-2 transcript than those patients without methylated TFPI-2 (P=0.04). Promoter hypermethylation of TFPI-2 is frequent and specific event in pediatric AML.

Over-expression of LAPTM4B is associated with poor prognosis and chemotherapy resistance in stages III and IV epithelial ovarian cancer.

BACKGROUND: The purpose of this study was to determine whether LAPTM4B over-expression is associated with the prognosis and chemotherapy resistance in patients with stages III and IV epithelial ovarian carcinoma, i.e., patients with peritoneal metastasis or lymph node metastasis of epithelial ovarian carcinoma. METHODS: LAPTM4B expression was evaluated in 10 normal ovarian and 113 stages III-IV ovarian carcinomas specimens by Western blotting analyses and immunohistochemistry. Univariate and multivariate analyses were performed to determine the association between LAPTM4B expression and prognosis and the relationship between LAPTM4B over-expression and chemotherapy resistance. RESULTS: Western blotting analysis demonstrated that LAPTM4B was overexpressed in ovarian cancers, and immunohistochemistry results revealed that 80 patients were LAPTM4B over-expression. The five-year overall survival (OS) rates for patients with high LAPTM4B expression and low LAPTM4B expression were 27.36% and 90.7%, respectively (hazard ratio = 20.611, 95% CI: 5.916-71.808, P < 0.0001). The five-year progression-free survival (PFS) rate was 17.68% for patients in the high-expression group and 84.42% for patients in the low-expression group (hazard ratio = 17.852, 95% CI: 6.31-5.935, P < 0.0001); The presence of chemotherapy resistance was significantly associated with LAPTM4B expression (OR: 36.609, 95% CI: 4.737-282.941, P = 0.0006). CONCLUSIONS: LAPTM4B over-expression is an independent factor in stages III-IV epithelial ovarian carcinoma prognosis and chemotherapy resistance, and it may be an important potential biomarker.

The active components derived from Penthorum chinensePursh protect against oxidative-stress-induced vascular injury via autophagy induction.

Oxidative stress-induced damage has been proposed as a major risk factor for cardiovascular disease and is a pathogenic feature of atherosclerosis. Although autophagy was reported to have a protective effect against atherosclerosis, its mechanism for reducing oxidative stress remains un-elucidated. In this study, we have identified 4 novel autophagic compounds from traditional Chinese medicines (TCMs), which activated the AMPK mediated autophagy pathway for the recovery of mitochondrial membrane potential (MMP) to reduce the production of reactive oxygen species (ROS) in Human umbilical vein endothelial cells (HUVECs). In this study, 4 compounds (TA, PG, TB and PG1) identified from Penthorum chinense Pursh (PCP) were demonstrated for the first time to possess binding affinity to HUVECs cell membranes via cell membrane chromatography (CMC) accompanied by UHPLC-TOF-MS analysis, and the 4 identified compounds induce autophagy in HUVECs. Among the 4 autophagic activators identified from PCP, TA (Thonningianin A, Pinocembrin dihydrochalcone-7-O-[3″-O-galloyl-4″,6″-hexahydroxydiphenoyl]-glucoside) is the major chemcial component in PCP, which possesses the most potent autophagy effect via a Ca2+/AMPK-dependent and mTOR-independent pathways. Moreover, TA efficiently reduced the level of ROS in HUVECs induced by H2O2. Additionally, the expression of pro- and cleaved-IL-1ß in the aortic artery of ApoE-KO mice were also alleviated at the transcription and post-transcription levels after the administration of TA, which might be correlated to the reduction of oxidative-stress induced inflammasome-related Nod-like receptor protein3 (NLRP3) in the aortic arteries of ApoE-KO mice. This study has pinpointed the novel autophagic role of TA in alleviating the oxidative stress of HUVECs and aortic artery of ApoE-KO mice, and provided insight into the therapeutic application of TA in treatment of atherosclerosis or other cardiovascular diseases.

Development of clinical mass spectrometry laboratories: opportunities and challenges / 中华检验医学杂志

Clinical application of mass spectrometry technology has attracted the attention of clinical laboratory experts due to its high sensitivity, high specificity, and capacities of simultaneous detection of multiple compounds. In recent years, mass spectrometry technology has made significant achievements in the fields of identification of pathogenic microorganism, detection of trace elements and heavy metals, small molecule hormones, vitamins, amino acids, peptides and proteins, as well as therapeutic drug monitoring (TDM) and poisoning drugs screening. In order to further clarify the opportunities and challenges brought by this complex mass spectrometry technology in the field of clinical laboratory, the Chinese Journal of Laboratory Medicine invited experts and scholars of laboratory medicine to share their experience and opinions on related items focusing on the positioning of mass spectrometry technology in the clinical laboratory, the development and improvement of the clinical laboratory by mass spectrometry technology, the challenges of interpreting mass spectrometry test results, the challenges of operating and managing clinical mass spectrometry laboratories, and ways of improving the application of clinical mass spectrometry laboratories with this technology. Agreement was achieved in that the introduction of mass spectrometry technology into the clinical laboratory could bring new directions and opportunities for clinical testing and research, and also is associated with a series of challenges such as the difficulty of sample pretreatment, the high cost and complexity of mass spectrometry technology, the complexity of data processing and interpretation, the lack of standards and norms, and the issue of determining the price of mass spectrometry examinations.

Identification and expression analysis of cellulose synthase family genes in Aquilaria sinensis / 药学学报

Cellulose synthase (CesA), one of the key enzymes in the biosynthesis of cellulose in plants, plays an important role in plant growth and plant resistance. In this study, a total of 21 AsCesA genes from Aquilaria sinensis were systematically identified and the physico-chemical characteristics were analyzed based on genome database and bioinformatical methods. The phylogenetic tree was constructed and the gene location on chromosome, cis-acting elements in the 2 000 basepairs upstream regulatory regions and conservative motifs were analyzed. The AsCesA proteins were mainly located on the plasma membrane. The number of amino acids of the proteins ranged from 390 to 1 261. The isoelectric point distributed from 5.67 to 8.86. All of the 21 AsCesA proteins possessed the transmembrane domains, the number of which was from 6 to 8. The genes were classified into 3 groups according to the phylogenetic relationship. Obvious differences were observed in motif composition in genes from different groups. However, motif2, motif6, motif7 and motif10 were observed in all of AsCesA proteins. Analysis of cis-acting elements indicated that AsCesA genes family has cis-acting elements related to plant hormones, abiotic stresses, and biological processes. Seven AsCesA genes with differential expression were selected according to the calli transcriptome data induced by NaCl at different times and their expression levels under different abiotic stresses were analyzed by quantitative real-time PCR. The results indicated that salt, low temperature, drought, and heavy metal stresses could affect the expression level of AsCesA genes, and the abundance of AsCesA1, AsCesA3 and AsCesA20 showed a significant change, implying their potential important roles to the abiotic stresses. The accumulation pattern of cellulose content under different abiotic stresses was similar to the expression trend of AsCesA genes. Our results provide valuable insights into the role of cellulose synthase in A.sinensis in plant defense.

Exploration on Acupuncture and Moxibustion Treatment Ideas for Gynecological Reproductive Diseases Based on the"Heart-kidney-Chong Ren-uterus"Reproductive Axis / 中国中医药信息杂志

This article mainly elaborated the acupuncture and moxibustion treatment scheme of"eighteen needles for reproduction"based on Professor You Zhaoling's reproductive axis theory of"heart-kidney-Chong Ren-uterus".The"eighteen needles for reproduction"aims to regulate the disordered reproductive axis in gynecological reproductive diseases.It selects the acupoints on the main viscera and meridians of the reproductive axis as the main acupoints,and the acupoints regulating the qi and blood of the related viscera as the matching acupoints.Through specific manipulation,it can regulate the qi and blood,dredge the meridians,and treat the viscera,so as to nourish the essence and help pregnancy,and provide ideas and reference for the treatment of gynecological reproductive diseases with acupuncture and moxibustion.

Evaluation and application of automated quality control of automatic pipeline in clinical biochemical and immunological detection / 中华检验医学杂志

Objective:To assess the applicability of fully automatic pipeline automated testing for internal quality control (automated quality control).Methods:Stability, assay efficiency and implementation costs of 18 biochemical tests, 5 immunoturbidimetric tests and 11 chemical illuminescent tests in the Department of Laboratory Medicine of Peking Union Hospital from January 2019 to July 2022 were evaluated using automated quality control implementation methods. The detailed method is as follows: quality control materials for biochemical, immunoturbidimetric and chemiluminescent tests were stored in the refrigerator in the pipeline which was controlled by the intermediate software, and were automatically retrieved and tested as pre-set followed by documenting and storing. The quality control setup for the biochemical tests included refreshing quality control materials daily and weekly,both of which were paralleled for 3 months. The on-line storage stability of quality control materials in the pipeline was evaluated by comparing the coefficients of variation ( CV) of the quality control results between the two patterns. Effect of automated quality control application was evaluated using 6 indicators, including the results′ variation of automatically performed and manually performed quality controls, the out-of-controlled rate, the consumption of quality control materials, the change of staff workload, the impact on the testing time of the first sample, and the failure rate of automated quality control. Results:(1) Storage stability of quality control materials in the pipeline: under the pattern of weekly refresh of the biochemical quality control materials, except for total carbon dioxide (TCO 2) (the CVs of low and high level quality control were respectively 20.24% and 21.82%) and sodium (the CV of low level quality control was 1.51%) that were greater than the allowable variation set by the laboratory, the CVs of the rest tests meet the lab requirements on the allowable variations. (2) The results′ variation of quality control in automatically performed and manually performed control patterns: in the patterns of daily refresh of biochemical quality control materials and weekly refresh of immunoturbidimetric and chemiluminescent quality control materials, the CVs of both low and high levels of quality control were lower in the automatically performed control pattern than that in manually performed pattern for 8 chemiluminescent items of dehydroepiandrosterone sulfate, estradiol, follicle stimulating hormone, luteinizing hormone, serum ferritin, serum folic acid, vitamin B12 and testosterone, 3 immunologic items of complement 3, C reactive protein and immunoglobulin G, and 10 biochemical items of alkaline phosphatase, glucose, calcium, chloride, potassium, lactate dehydrogenase, sodium, urea, low density lipoprotein cholesterol, and adenosine deaminase. The out-of-control rates of biochemistry, immunoturbidimetric and chemiluminescence tests in both quality control patterns conformed with the clinical routine work requirements. (3) Comparison of quality control materials′ consumption: compared with manually performed quality control, weekly consumption of automatically performed chemiluminescent quality control materials decreased 37.5% (from 8 ml to 5 ml); weekly consumption of automatically performed immunoturbidimetric quality control materials decreased 33.3% (from 3 ml to 2 ml). (4)Comparison of staff workload and first sample testing time: compared with manually performed quality control, automatical quality control reduced manual work by about 156 steps per week, and the daily initial testing time was earlier by 15 min on average. The failure rate was 54.5% (37/64) during the early-stage application of the automated quality control which dropped to 10.2% (13/128) in the late-stage. Conclusion:The results of automated quality control detected in the pipeline system meet the quality indicators′ requirements of the laboratory, and the application of automated quality control can improve the quality control, save costs, reduce workload, and improve work efficiency.

Advances in therapeutic drug monitoring methods based on liquid chromatography-tandem mass spectrometry / 中华检验医学杂志

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology has the characteristics of high specificity and high throughput, making it rapidly applied and developed in the field of clinical testing. Its application in the monitoring of therapeutic drugs can effectively improve the quantitative accuracy and sensitivity, and formulate a personalized and optimal dosing plan for patients. However, this technology still faces some challenges, and automation, quality control, and quantitative traceability will be the future development direction.

Comparison of Direct and Extraction Immunoassay Methods With Liquid Chromatography-Tandem Mass Spectrometry Measurement of Urinary Free Cortisol for the Diagnosis of Cushing’s Syndrome

Background@#Twenty-four-hour urinary free cortisol (UFC) measurement is the initial diagnostic test for Cushing’s syndrome (CS). We compared UFC determination by both direct and extraction immunoassays using Abbott Architect, Siemens Atellica Solution, and Beckman DxI800 with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, we evaluated the value of 24-hr UFC measured by six methods for diagnosing CS. @*Methods@#Residual 24-hr urine samples of 94 CS and 246 non-CS patients were collected.A laboratory-developed LC-MS/MS method was used as reference. UFC was measured by direct assays (D) using Abbott, Siemens, and Beckman platforms and by extraction assays (E) using Siemens and Beckman platforms. Method was compared using Passing–Bablok regression and Bland–Altman plot analyses. Cut-off values for the six assays and corresponding sensitivities and specificities were calculated by ROC analysis. @*Results@#Abbott-D, Beckman-E, Siemens-E, and Siemens-D showed strong correlations with LC-MS/MS (Spearman coefficient r = 0.965, 0.922, 0.922, and 0.897, respectively), while Beckman-D showed weaker correlation (r = 0.755). All immunoassays showed proportionally positive bias. The areas under the curve were 0.975 for Abbott-D, 0.972 for LCMS/MS, 0.966 for Siemens-E, 0.948 for Siemens-D, 0.955 for Beckman-E, and 0.877 for Beckman-D. The cut-off values varied significantly (154.8–1,321.5 nmol/24 hrs). Assay sensitivity and specificity ranged from 76.1% to 93.2% and from 93.0% to 97.1%, respectively. @*Conclusions@#Commercially available immunoassays for measuring UFC show different levels of analytical consistency compared to LC-MS/MS. Abbott-D, Siemens-E, and Beckman-E have high diagnostic accuracy for CS.

Dynamic protein expression of NF-κB following rat intracerebral hemorrhage and its association with apoptosis.

The aim of the present study was to evaluate the dynamic protein expression of nuclear factor (NF)-κB and apoptosis in the cerebral tissue surrounding hematoma following intracerebral hemorrhage (ICH) in rats. A total of 80 healthy male Wistar rats were divided into a sham-surgery group and an ICH group. The ICH model was established by injecting autogenous non-heparin anticoagulant arterial blood into the caudate putamen. NF-κB levels were assessed by immunohistochemistry at different time points subsequent to surgery, and apoptosis condition was investigated by terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling. Different levels of NF-κB were expressed in the cerebral tissue around the ICH at each time point in the ICH group. NF-κB protein expression was detected at 3 h following hemorrhage, mainly in the cytoplasm. Following 6 h, NF-κB was identified in the nucleus. Its expression peaked at 72 h following hemorrhage, and persisted for 5 days. Apoptosis was observed 6 h following hemorrhage, and had increased significantly by 12 h. The rate of apoptosis continued to rise from 72-120 h following hemorrhage. Correlation analysis revealed a significant positive correlation between NF-κB expression and apoptosis (r=0.753; P<0.01). The enhancement of NF-κB expression and apoptosis around ICH, and the significant positive correlation between NF-κB expression and apoptosis, indicates that NF-κB activation may enhance cerebral apoptosis in rats following ICH.

Experimental study of PET apoptotic probe 18F-1 in monitoring radiotherapy response of triple-negative breast cancer / 中华核医学与分子影像杂志

Objective:To explore the application potential of 18F-Asp-Glu-val-Asp (DEVD)-Cys(StBu)-PPG(CBT)-AmBF 3 ( 18F-1; PPG: propargyl-glycine; CBT: 2-cyanobenzothiazole; AmBF 3: ammoniomethyl-trifluoroborate) PET imaging in early monitoring of triple-negative breast cancer (TNBC) radiotherapy response. Methods:Ten MDA-MB-231 tumor bearing nude mice models were constructed and divided into radiotherapy group ( n=5) and non-radiotherapy group ( n=5) by random sampling method. The radiotherapy group was treated with single irradiation at a dose of 8 Gy. 18F-1 microPET imaging was performed in the radiotherapy and non-radiotherapy groups, and the tumor uptake and muscle uptake in 2 groups at different time points (2.5, 7.5, 12.5, 17.5, 22.5, 27.5, 32.5, 37.5, 42.5, 47.5, 52.5, 57.5 min after injection) were analyzed. The specific uptake of the probe in apoptotic cells was verified by radioautography, HE staining and immunofluorescent staining. Repeated measures analysis of variance and one-way analysis of variance were used to analyze data. Results:18F-1 microPET imaging showed that there was significant difference between tumor uptake and muscle uptake in radiotherapy group ( F=20.27, P=0.011). The uptake of radiotherapy group was the highest at 7.5 min after injection ((4.64±0.35) percentage activity of injection dose per gram of tissue(%ID/g)). There was no significant difference between tumor uptake and muscle uptake in the non-radiotherapy group ( F=1.81, P=0.215). The tumor/muscle (T/M) ratio of radiotherapy group was higher than that of non-radiotherapy group ( F=31.95, P=0.005), with the highest at 47.5 min after injection (2.49±0.46). Radioautography showed that the tumor radioactivity in radiotherapy group was higher than that of muscle in radiotherapy group, and was also higher than tumor and muscle radioactivies in non-radiotherapy group ( F=116.79, P<0.001). HE staining and immunofluorescent staining verified that 18F-1 could specifically detect the activity of caspase-3 activated in tumor cells after radiotherapy. Conclusion:18F-1 can specifically recognize the activated caspase-3 after TNBC radiotherapy, and monitor radiotherapy response at the molecular level by apoptosis PET imaging.

Parathyroid hormone promotes osteoblastic differentiation of endothelial cells via the extracellular signal-regulated protein kinase 1/2 and nuclear factor-κB signaling pathways.

Vascular calcification (VC) occurs in patients with chronic kidney disease (CKD) and contributes to cardiovascular dysfunction and mortality. Parathyroid hormone (PTH) is a crucial regulator of VC. High PTH serum levels constitute as a major risk factor for patients with CKD. However, the effect and mechanism of PTH on osteoblastic differentiation in endothelial cells have not been fully elucidated. In the present study, the role of PTH in VC was investigated using an in vitro calcification model. Endothelial cells were stimulated with PTH in the femto- to picomolar range. As determined by western blot analysis and ELISA, osteoblastic differentiation, as indicated by the BMP2 marker, occurred with maximum effect at 1×10-10 mmol/l PTH. The results indicate that PTH promotes osteoblastic differentiation of endothelial cells, as demonstrated by the increased expression of bone morphogenetic protein (BMP) 2 and BMP4. In addition, western blot analysis revealed that PTH activated the extracellular signal-regulated protein kinase (Erk)1/2 and nuclear factor (NF)-κB signaling pathways. However, reverse transcription-quantitative polymerase chain reaction demonstrated that inhibitors specific to Erk1/2 and NF-κB eradicated the effect of PTH treatment on BMP2, BMP4, ALP and RUNX2 expression. These results demonstrate that PTH promotes the osteoblastic differentiation of endothelial cells via the Erk1/2 and NF-κB signaling pathways, which suggests a potential role of PTH in the promotion of VC. These findings provide an insight into the association between PTH and cardiovascular disease.

Homocysteine level of Tibetan population settled down at different altitudes / 基础医学与临床

Objective To investigate and analyze the level of homocysteine(Hcy)in Tibet and to analyze the differences of Hcy level in different altitude regions,genders and ages,and thus to provide the prevalence profile of hyperhomocysteine and the differences in relevant tests between HHcy(hyperhomocysteinemia)and non-HHcy pop-ulations.Methods Totally 1 615(male n＝585)subjects were selected from Ngari,Lhasa,Shigatse and Nyingchi plat-eau areas of Tibet by stratified cluster sampling.Serum Hcy level was analyzed and the difference of Hcy level in pop-ulations located at different altitude plateau areas,gender groups were found.The prevalence of hyperhomocysteine and related test were analyzed.Kruskal Wallis test was used to compare Hcy levels in different altitudes,genders and age groups,and Pearson Chi-square test was used to compare HHcy prevalence.Variance analysis was used for the differences of different test indicators between non-HHcy and HHcy populations.Results The level of Hcy in differ-ent regions and different genders were statistically significant,which was higher in males than that in females,and higher in Lhasa and Shigatse than in Nyingchi and Ngari.There was difference in serum HHcy prevalence among dif-ferent genders,regions and age groups.Males showed a higher level than females,people from Lhasa and Shigatse showed a higher level than those from Nyingchi and Ngari.Conclusions The incidence of hyperhomocysteinemia in Tibet is statistically significant in different areas,different genders and different age groups.So this study provides a scientific basis for the rational use of Hcy as an indicator in clinical practice of prevention and treatment of related diseases in plateau areas.

Explore the reasons affecting the consistency of reference intervals established by two types of indirect methods for 34 biochemical analytes / 中华检验医学杂志

Objective:To compare the differences of reference intervals (RI) established by two types of indirect methods for 34 biochemical analytes, and to explore the possible factors that affect the consistency of the two methods.Methods:This was a retrospective study. Based on data of albumin (Alb), alkaline phosphatase (ALP), alanine aminotransferase (ALT), apolipoprotein A1(ApoA1), ApolipoproteinB (ApoB), aspartate aminotransferase (AST), calcium (Ca), cholinesterase (ChE), chloride (Cl), creatinine (Cr), high-sensitivity C-reactive protein (hsCRP), Cystatin (CysC), direct bilirubin (DBil), free fatty acid (FFA), glycated albumin(GA), gamma-glutamyltransferase (GGT), glucose (Glu), high density lipoprotein cholesterol (HDL-C), potassium (K), lactate dehydrogenase (LD), low density lipoprotein cholesterol (LDL-C), lipoprotein a [Lp (a)], sodium (Na), phosphorus (P), prealbumin (PA), superoxide dismutase (SOD), total bile acid (TBA), total bilirubin (TBil), total cholesterol (TC), total carbon dioxide (TCO 2), triglyceride (TG), total protein (TP), uric acid (UA) and urea (UR) of individuals who underwent physical examination at Peking Union Medical College Hospital from January 1, 2018 to December 31, 2019, Box-Cox algorithm was used to improve the data distribution and Tukey method was used to identify outliers. Variance component model was established, and standard deviation ratio (SDR) was calculated to determine whether the RIs of 34 biochemical analytes should be established according to age or sex The non-parametric method and kosmic algorithm were used to establish the RIs and 90% confidence intervals (CIs) of 34 biochemical analytes, and the coincidence of the 90% CIs of the reference limits for two methods was compared. Results:The skewness coefficients of ALP(male, female18-59), ALT, AST, hsCRP, DBil, GGT, Lp (a), TBA, TBil, TG, Glu, HDL-C(male) and CysC, GA, UR in the elderly group deviated from 0, and their kurtosis coefficients also deviated from 3. For these biochemical analytes, the point estimates of the RIs established by the two methods differed greatly and the 90% CIs did not overlap. The analytes with good normality were Alb, ApoA1, ApoB, Ca, ChE, Cl, Cr(E), CysC(18-59), FFA, GA(18-59), HDL-C(female), K, LDL-C, Na, P, PA, SOD, TC, TCO 2, TP and UR. The consistency is good. Except for Ca, 90% CIs of reference limits for some analyte between the two methods coincide with each other. Conclusions:The consistency of different indirect methods is affected by the normality of data.

Establishing and evaluating a robust method based on LC-MS/MS for simultaneous determination of Aβ1-42,Aβ1-40 and A β1-38 in cerebrospinal fluid / 中华检验医学杂志

Objective:To establish and validate an LC-MS/MS method for simultaneous determination of Aβ 1-42, Aβ 1-40, and Aβ 1-38 in cerebrospinal fluid. Additionally, the consistency between this method and three mainstream detection methods was evaluated.Methods:This study involved method establishment, validation, and consistency evaluation. The N15 labeled β-amyloid protein was used as the internal standard. Extraction was performed using Waters MCX 96-wells solid phase extraction plate, and the eluent was collected to QuanRecovery MaxPeak 700 μl plate. At the positive ion mode, the multi-reaction ion monitoring mode based on electric spray ionization is chosen for the determination of CSF Aβ 1-42, Aβ 1-40, and Aβ 1-38. Referring to the CLSI C62-A and EP-15A3 guidelines, the method is evaluated and verified, including quantitation of limit (LOQ), linearity, recovery, precision, and accuracy. In addition, a total of 57 clinical residual CSF samples were collected and the concentrations of Aβ 1-42 and Aβ 1-40 were determined based on manual INNOTEST ELISA assay and Lumipulse G and Roche Elecsys fully automated biochemical analyzers. The comparison analysis and deviation evaluation were conducted by passing-bablok and Bland Altman methods.Results:The analysis time of this method is 8 min, and the LOQ of Aβ 1-42, Aβ1-40 and Aβ1-38 is 0.1 ng/ml, 0.5 ng/ml, and 0.1 ng/ml, respectively, and the linear range can meet the needs of clinical detection. Respectively, the recovery is 86.2%-93.8%, 100.9%-103.9% and 103.3%-107.1%; the total imprecision is 4.7%-7.4%, 3.5%-4.6% and 5.2%-10.9%. The measured values of Aβ 1-42 certified reference materials are all within the allowable uncertainty requirements. Moreover, the carryover rate of three analytes was all≤0.11%. In addition, the correlations of Aβ 1-42 and Aβ1-40 in CSF between this LC-MS/MS method and the INNOTEST ELISA method, Lumipulse G and Roche Elecsys fully automated biochemical analyzers were all deemed good, with correlation coefficient (r) ranging from 0.920 to 0.970. However, the measured values between the four methods were remarkably different.Conclusion:We established and validated a robust method based on LC-MS/MS technology for simultaneous determination of Aβ 1-42, Aβ 1-40, and Aβ 1-38 in CSF. The method is accurate, simple, and suitable for clinical measurements. However, despite good correlations, there were substantial differences in the measurement results of Aβ 1-42 and Aβ 1-40 among different analytical platforms, indicating the need for further promotion of harmonization and standardization processes for AD classic biomarkers.

Cloning, expression analysis and enzyme activity verification of dihydroflavonol 4-reductase from Cistanche tubulosa (Schenk) Wight flower / 药学学报

Dihydroflavonol 4-reductase (DFR) plays an essential role in the biosynthesis of anthocyanin and regulation of plant flower color. Based on the transcriptome data of Cistanche tubulosa (Schenk) Wight, a full-length cDNA sequence of CtDFR gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR). CtDFR contains an open reading frame (ORF) of 1 263 bp which encodes 420 amino acids with a predicted molecular weight of 47.5 kDa. The sequence analysis showed that CtDFR contains a nicotinamide adenine dinucleotide phosphate (NADPH) binding domain and a specific substrate binding domain. The expression analysis indicated that CtDFR was highly expressed in red and purple flowers, and the relative expression levels were 4.04 and 19.37 times higher than those of white flowers, respectively. The recombinant CtDFR protein was expressed in E.coli BL21 (DE3) using vector pET-28a-CtDFR and was purified. In vitro enzyme activity analysis, CtDFR could reduce three types of dihydroflavonols including dihydrokaempferol, dihydroquercetin, and dihydromyricetin to leucopelargonidin, leucocyanidin and leucodelphinidin. Subcellular localization analysis showed that CtDFR was mainly localized in the cytoplasm. These results demonstrate that CtDFR plays an important role in regulation of flower color in C. tubulosa and make a valuable contribution for the further investigation on the regulation mechanism of C. tubulosa (Schenk) Wight flower color.