Investigation and Analysis of Reference Intervals for Blood Cell Parameters in a Healthy Population from Daxingan Region Inner Mongolia.

BACKGROUND: Blood cell parameters in a healthy population of different ages and genders from the Daxingan region of Inner Mongolia were evaluated to establish reference intervals (RIs) for venous blood parameters for healthy individuals from this region. METHODS: Venous blood specimens were collected from 1757 healthy individuals aged 7-65 years and analyzed by an XE-5000 automated hematology analyzer. Results were statistically analyzed by gender and age group. RIs for venous blood parameters of children were compared to the National Clinical Laboratory Procedures and those of adults were compared to the Health Industry Standards of the People's Republic of China (WST405-2012). RESULTS: In the Daxingan region, WBC, RBC, MCV, and NEUT%, MONO%, and EO% of healthy children were significantly different between genders (p < 0.05). Other parameters were not significantly different (p > 0.05). All parameters of adults were significantly different between different genders (p < 0.05) except for LYMPH% and BASO%. Comparison of mean values between children and adults of the same gender revealed significant differences in each blood cell parameter (p < 0.05). RBC, MCV, MCH, MCHC, PLT, MONO%, and BASO% of adult men were significantly different between different age groups (p < 0.05), and HGB, HCT, and EO% of adult women were significantly different between different age groups (p < 0.05). For children, only the mean of MCHC and EO% were close to the National Clinical Laboratory Procedures. Mean of RBC, HGB, PLT, LYMPH%, and MONO% were higher and the remaining parameters were lower than the National Clinical Laboratory Procedures. Compared with WST405-2012 for adults, PLT of women aged 18-40 years and NEUT% of adults were higher, whereas the mean of EO% and BASO% were significantly lower. CONCLUSIONS: RIs for venous blood parameters change with age, geographic region, ethnic group, and gender. There is a great necessity to establish RIs for venous blood parameters among the healthy population in the Daxingan region.

Reference Intervals for Routine Blood Tests in 468 Healthy Mongolian Children.

BACKGROUND: To establish reference intervals for routine blood parameters of healthy Mongolian children in the Inner Mongolia Daxingan region. METHODS: Venous blood samples from 468 children aged 7 - 13 years were analyzed with a Sysmex XE-5000 Automated Hematology Analyzer. Statistical analysis was used to compare findings with the National Clinical Laboratory Procedures and to analyze according to gender and ethnic group. RESULTS: The reference range for each blood cell parameter is as follows: WBC (x 109/L): 3.44 - 9.27 for males, 3.79 - 9.79 for females; RBC (x 1012/L): 4.25 - 5.36 for males, 4.20 - 5.21 for females; HGB (g/L): 118 - 149. HCT (%): 34.9 - 43.4; MCV (fl): 75.1 - 88.2 for males, and 76.6 - 90.6 for females; MCH (pg): 25.5 - 30.3 for males, and 25.7 - 31.0 for females; MCHC (g/L): 327 - 356 for males, 323 - 353 for females; PLT (109): 109 - 209; NEUT %: 34.6 - 71.9; LYMPH %: 19.8 - 56.8; MONO %: 2.8 - 9.5 for males, and 3.1 - 7.9 for females; EO %: 0.0 - 3.6; BASO %: 0.0 - 1.0 for males, and 0.0 - 0.9 for females. There were significant differences between genders with respect to the WBC, RBC, MCV, MCH, MCHC, MONO %, and BASO % parameters (p < 0.05), but not other parameters (p > 0.05). CONCLUSIONS: The blood parameters of children can vary with geography, nationality, gender, and other variables. Therefore, it is necessary to obtain reference intervals for Mongolian children according to their gender and age.

Reference intervals for six lipid analytes in 8-14 year-old school children from three different ethnical groups in the Hulun Buir area of China.

BACKGROUND: Reference intervals vary according to gender, age, ethnicity, diet, and other factors. It is therefore recommended that population-specific reference intervals be established. This study investigated reference intervals of blood fat of healthy primary students (8 - 14 years) from Mongolian, Ewenki, and Han ethnicities in Hulun Buir area. METHODS: Blood samples were collected from 1,723 children aged 8 - 14 years: 805 boys (46%) and 918 girls (54%) were analyzed for cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A1 (APOAI), and apolipoprotein B (APOB) levels. TC and LDL-C 90 and 75 percentiles were considered as the critical high lipoprotein level and the lipoprotein concentration standard, TG 90 percentiles as high blood triglycerides concentrations, 5 percentiles as HDL-C reference range lower level and 95 percentiles as reference range of APOAI and APOB, the normal lipid reference interval for three ethnic groups of pupils were set up. RESULTS: There were significant differences between Han and other ethnicities with respect to TC, TG, HDL-C, APOAI, APOB (p < 0.01), but not LDL-C (p > 0.05). There were significant differences in Mongolian and Ewenki ethnicities with respect to LDL-C, HDL-C, APOAI and APOB (p < 0.01), but not TC, TG (p > 0.05). There was significant difference between boys and girls of Han and Mongolian ethnicities in TG, HDL-C, LDL-C, APOAI, APOB lipid levels (p < 0.01); and there was significant difference between boys and girls of Ewenki ethnicity with respect to TG, HDL-C, APOAI, APOB lipid levels (p < 0.01). CONCLUSIONS: Reference intervals of serum lipid parameters blood fat for healthy Mongolian, Ewenki, and Han ethnicities of primary students in Hulun Buir are presented, which provide an important update for lipid markers and suggest earlier incidence of hypercholesterolemia when comparing to previous ranges.

Proteomic and metabolomic analysis of the serum of patients with tick-borne encephalitis.

Tick-borne encephalitis virus (TBEV) is a common virus in Europe and Asia, causing around 10,000 to 10,500 infections annually. It affects the central nervous system and poses threats to public health. However, the exact molecular mechanisms of TBE pathogenesis are not yet fully understood due to the complex interactions between the virus and its host. In this study, a comprehensive analysis was conducted to characterize the serum metabolome and proteome of adult patients infected with TBEV, in comparison to a control group of healthy individuals. Liquid chromatography tandem mass spectrometry (LC-MS) was employed to monitor metabolic and proteomic alternations throughout the progression of the disease, significant physiological changes associated with different stages of the disease were identified. A total of 44 proteins and 115 metabolites exhibited significantly alternations in the sera of patients diagnosed with TBE. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses of these metabolites and proteins revealed differential enrichment of genes associated with the extracellular matrix, complement binding, hemostasis, lipid metabolism, and amino acid metabolism between TBE patients and healthy controls. We gained valuable understanding of the specific metabolites implicated in the host's responses to TBE, establishing a basis for further research on TBE disease. SIGNIFICANCE: The current investigation revealed a comprehensive and systematic differences on TBE using LC-MS platform from human serum samples of TBE patients and healthy individuals providing the immune response to the invasion of TBE.

A novel rapid visual nucleic acid detection technique for tick-borne encephalitis virus by combining RT-recombinase-aided amplification and CRISPR/Cas13a coupled with a lateral flow dipstick.

Tick-borne encephalitis virus (TBEV), a zoonotic pathogen, can cause severe neurological complications and fatal outcomes in humans. Early diagnosis of TBEV infection is crucial for clinical practice. Although serological assays are frequently employed for detection, the lack of antibodies in the early stages of infection and the cross-reactivity of antibodies limit their efficacy. Conventional molecular diagnostic methods such as RT-qPCR can achieve early and accurate identification but require specialized instrumentation and professionals, hindering their application in resource-limited areas. Our study developed a rapid and visual TBEV molecular detection method by combining RT-recombinase-aided amplification, the CRISPR/Cas13a system, and lateral flow dipsticks. The diagnostic sensitivity of this method is 50 CFU/ml, with no cross-reactivity with a variety of viruses. The detection can be carried out within 1 h at a temperature between 37 and 42 °C, and the results can be visually determined without the need for complex instruments and professionals. Subsequently, this assay was used to analyze clinical samples from 15 patients suspected of TBEV infection and 10 healthy volunteers, and its sensitivity and specificity reached 100 %, which was consistent with the results of RT-qPCR. These results indicate that this new method can be a promising point-of-care test for the diagnosis of tick-borne encephalitis.

TRIM37 interacts with PTEN to promote the growth of human T-cell acute lymphocytic leukemia cells through regulating PI3K/AKT pathway.

Background: TRIM37 has been reported to be associated with the tumorigenesis of cancers. However, the role of TRIM37 in T-cell acute lymphoblastic leukemia (T-ALL) remains unclear. This study aimed to characterize the effect of TRIM37 on T-ALL. Methods: TRIM37 expression in T-ALL patients and T-ALL cell lines was determined by qRT-PCR and Western blot. Knockdown or overexpression of TRIM37 was conducted by transferring small-interfering TRIM37 or lentivirus-mediated transducing into T-ALL cells. CCK-8 assay and flow cytometry assay were conducted to analyze the proliferation and apoptosis of T-ALL cells. Co-immunoprecipitation experiments were conducted to investigate the relationship between TRIM37 and PTEN and the ubiquitination of PTEN. Results: Our results suggested that TRIM37 expression was upregulated in the blood of T-ALL patients and T-ALL cell lines. Knockdown of TRIM37 noticeably inhibited the proliferation and promoted apoptosis of T-ALL cells. Ectopic expression of TRIM37 promoted the proliferation and suppressed the apoptosis rate of MOLT-4 cells and enhanced the phosphorylation of AKT. Moreover, TRIM37 interacted with PTEN and accelerated the degradation of PTEN via TRIM37-mediated ubiquitination in T-ALL cells. Moreover, TRIM37 reduced the sensitivity of T-ALL cells to bortezomib treatment. Additionally, PI3K/AKT signaling pathway was involved in the function of TRIM37 in T-ALL. TRIM37 contributed to the proliferation of T-ALL cells and reduced the susceptibility of T-ALL cells to bortezomib treatment through ubiquitination of PTEN and activating PI3K/AKT signaling pathway. Conclusions: Our study suggested that TRIM37 could be considered as a therapeutic target for T-ALL.

Serum Metabolomics of Tick-Borne Encephalitis Based on Orbitrap-Mass Spectrometry.

BACKGROUND: Tick-borne encephalitis virus (TBEV), the most prevalent arbovirus, causes potentially fatal encephalitis in humans. Prevalent in northeast China, tick-borne encephalitis (TBE) poses a major threat to public health, local economies and tourism. There are no biomarkers for TBE, which is classified serologically and clinically. Due to sample heterogeneity of samples and different detection platforms, obtaining stable markers is a great challenge for metabolomics. Accurate annotation is vital for data mining and interpretation. OBJECTIVE: To identify reliable biomarkers of TBEV infection. METHODS: An untargeted metabolomics analysis of serum from 30 TBE patients and 30 healthy controls was carried out. Liquid chromatography-mass spectrometry (LC-MS)-based metabolomics methods were used to characterize the subjects' serum metabolic profiles and to screen and validate TBE biomarkers. RESULTS: A total of 3370 molecular features were extracted from each sample, and the peak intensity of each feature was obtained. Pattern analysis, principal component analysis, partial least squares discriminant analysis were used to screen for potential metabolites. Bilirubin, LysoPC (18:1[9Z]), palmitic acid, and CL (8:0/8:0/8:0/8:0) were significantly different. Pathway enrichment analysis showed that these metabolites were in the fatty acid biosynthesis and glycerophospholipid metabolism pathways. The phospholipid family had a significant difference in both the difference ratio and the abundance. CONCLUSION: Phospholipids may be used to distinguish TBEV patients from healthy controls. TBEV infection affects the normal metabolic activity of host cells, providing insight into the pathogenesis of TBE.

Insights into the molecular basis of tick-borne encephalitis from multiplatform metabolomics.

BACKGROUND: Tick-borne encephalitis virus (TBEV) is the most prevalent arbovirus, with a tentative estimate of 10,000 to 10,500 infections occurring in Europe and Asia every year. Endemic in Northeast China, tick-borne encephalitis (TBE) is emerging as a major threat to public health, local economies and tourism. The complicated array of host physiological changes has hampered elucidation of the molecular mechanisms underlying the pathogenesis of this disease. METHODOLOGY/PRINCIPLE FINDINGS: System-level characterization of the serum metabolome and lipidome of adult TBEV patients and a healthy control group was performed using liquid chromatography tandem mass spectrometry. By tracking metabolic and lipid changes during disease progression, crucial physiological changes that coincided with disease stages could be identified. Twenty-eight metabolites were significantly altered in the sera of TBE patients in our metabolomic analysis, and 14 lipids were significantly altered in our lipidomics study. Among these metabolites, alpha-linolenic acid, azelaic acid, D-glutamine, glucose-1-phosphate, L-glutamic acid, and mannose-6-phosphate were altered compared to the control group, and PC(38:7), PC(28:3;1), TAG(52:6), etc. were altered based on lipidomics. Major perturbed metabolic pathways included amino acid metabolism, lipid and oxidative stress metabolism (lipoprotein biosynthesis, arachidonic acid biosynthesis, leukotriene biosynthesis and sphingolipid metabolism), phospholipid metabolism and triglyceride metabolism. These metabolites were significantly perturbed during disease progression, implying their latent utility as prognostic markers. CONCLUSIONS/SIGNIFICANCE: TBEV infection causes distinct temporal changes in the serum metabolome and lipidome, and many metabolites are potentially involved in the acute inflammatory response and immune regulation. Our global analysis revealed anti- and pro-inflammatory processes in the host and changes to the entire metabolic profile. Relationships between metabolites and pathologies were established. This study provides important insight into the pathology of TBE, including its pathology, and lays the foundation for further research into putative markers of TBE disease.

CX3CL1 increases invasiveness and metastasis by promoting epithelial-to-mesenchymal transition through the TACE/TGF-α/EGFR pathway in hypoxic androgen-independent prostate cancer cells.

Epithelial-to-mesenchymal transition (EMT) endows cancer cells with enhanced invasive and metastatic potential during cancer progression. Fractalkine, also known as chemokine (C-X3-C motif) ligand 1 (CX3CL1), the only member recognized so far that belongs to the CX3C chemokine subfamily, was reported to participate in the molecular events that regulate cell adhesion, migration and survival of human prostate cancer cells. However, the relationship between CX3CL1 and EMT remains unknown. We treated DU145 and PC-3 cells with CX3CL1 under hypoxic conditions. The migration and invasion abilities of DU145 and PC-3 cells were detected by Transwell assays. Induction of EMT was verified by morphological changes in the DU145 and PC-3 cells and analysis of protein expression of EMT markers such as E-cadherin and vimentin. To identify the involved signaling pathway in CX3CL1-induced EMT, activation of epidermal growth factor receptor (EGFR) was measured using western blot analysis, and Slug expression was detected with or without an EGFR inhibitor prior to CX3CL1 treatment. Concentrations of soluble and total TGF-α in the CX3CL­treated DU145 cells were detected by ELISA. Additionally, we determined the involvement of the TACE/TGF-α/EGFR pathway in CX3CL1­induced EMT using RNA interference and specific inhibitors. CX3CL1 increased the migration and invasiveness of the DU145 and PC-3 cells, and resulted in characteristic alterations of EMT. Our results demonstrated that TACE/TGF-α/EGFR pathway activation and subsequent upregulation of Slug expression were responsible for CX3CL1­induced EMT, and contributed to the migration and inva-sion of prostate cancer cells. Inhibition of TACE/TGF-α/EGFR signaling reversed EMT and led to reduced migration and invasion abilities of the prostate cancer cells. We provide initial evidence that CX3CL1 exposure resulted in EMT occurrence and enhancement of cell migration and invasion through a mechanism involving activation of TACE/TGF-α/EGFR signaling. These findings revealed that CX3CL1 may serve as a new target for the treatment of prostate cancer.

Upregulation of fractalkine contributes to the proliferative response of prostate cancer cells to hypoxia via promoting the G1/S phase transition.

Hypoxia is a common phenomenon in prostate cancer, which leads to cell proliferation and tumor growth. Fractalkine (FKN) is a membrane­bound chemokine, which is implicated in the progression of human prostate cancer and skeletal metastasis. However, the association between FKN and hypoxia­induced prostate cancer cell proliferation remains to be elucidated. The present study demonstrated that hypoxia induced the expression and secretion of FKN in the DU145 prostate cancer cell line. Furthermore, inhibiting the activity of FKN with the anti­FKN FKN­specific antibody markedly inhibited hypoxia­induced DU145 cell proliferation. Under normoxic conditions, DU145 cell proliferation markedly increased following exogenous administration of human recombinant FKN protein, and the increase was significantly alleviated by anti­FKN, indicating the importance of FKN in DU145 cell proliferation. In addition, subsequent determination of cell cycle distribution and expression levels of two core cell cycle regulators, cyclin E and cyclin­dependent kinase (CDK)2, suggested that FKN promoted the G1/S phase transition by upregulating the expression levels of cyclin E and CDK2. The results of the present study demonstrated that hypoxia led to the upregulation of the secretion and expression of FKN, which enhanced cell proliferation by promoting cell cycle progression in the prostate cancer cells. These findings provide evidence of a novel function for FKN, and suggest that FKN may serve as a potential target for treating androgen­independent prostate cancer.

DNA methylation inhibitor, decitabine, promotes MGC803 gastric cancer cell migration and invasion via the upregulation of NEDD4­1.

Gastric cancer is the fourth most common cancer type and the second leading cause of cancer­associated mortality worldwide. Metastasis is a crucial feature of its progression. DNA methylation provides a key epigenetic signature in the epigenetic regulation pathway, and is implicated in transcriptional regulation. CpG sites, which are associated with gene transcriptional activity, are underrepresented in the mammalian genome and tend to be clustered within CpG islands (CGIs) located in the vicinity of the transcription start sites of the majority of the protein­coding genes in humans. The DNA methylation inhibitor, decitabine (DAC), has been demonstrated to be active in hematological disorders. The majority of previous studies in cancer cells demonstrated that DAC inhibits cell proliferation and the motility of the cells. However, since demethylation across the entire genome alters the expression of a large number of genes, the effects of DAC in different tumor cell types are difficult to accurately predict. Neural precursor cell­expressed, developmentally downregulated (NEDD)4­1, a member of the NEDD4 family, which belongs to the E3­ubiquitin ligase family, was reported to be highly expressed in a wide range of tumor types, and it activates the phosphoinositide 3­kinase/Akt pathway by degrading phosphatase and tensin homolog. NEDD4­1 promotes the migration and invasion of glioma cells via the ubiquitination and subsequent degradation of cyclic nucleotide­Ras guanine nucleotide exchange factors (CNrasGEFs). In gastric cardia adenocarcinoma, NEDD4­1 acts as an exceptional prognostic biomarker. In the present study, DAC was revealed to promote the invasive properties of MGC803 gastric cancer cells. NEDD4­1 targeted the CNrasGEF­mediated DAC invasion­promoting activity in MGC803 cells, and CGI methylation in neither the NEDD4 promoter nor the first intron was demonstrated to be associated with this effect. The results of the present study revealed that DAC exerts variable effects in different gastric cancer cell lines and may provide a reference for DAC administration in the clinic.