Harmonization of distributed multi-center analysis based on dried blood spot reference materials supporting the screening of neonatal inherited metabolic disorders.

BACKGROUND: The standardization of quantification data is critical for ensuring the reliability and measurement traceability in the screening of neonatal inherited metabolic disorders. However, the availability of national certified reference materials is limited in China. METHODS: In this study, we developed a series of dried blood spot (DBS) reference materials containing 9 amino acids (AA) and 10 acylcarnitines (AC) for neonatal screening. Four levels of the reference materials were measured with tandem mass spectrometry (MS/MS) by seven laboratories using different commercial In Vitro Diagnostic Device (IVD) kits. Then, 100 clinical samples were measured using both derivatization and non-derivatization methods by the same laboratory. RESULTS: We found high homogeneity and stability at all levels of the reference materials, with the coefficient of variation (CV) of the analytes less than 15%. These reference materials can be used to assess the testing capabilities of different laboratories. Our test also revealed that the correction factors (CF) calculated by the reference materials, along with clinical samples, could increase the consistency for different kits. CONCLUSION: The DBS reference materials proposed in this study provide reliability for the harmonization in multi-center analysis for the screening of neonatal inherited metabolic disorders. And applying our correction method for the screening could improve the data consistency of the DBS samples prepared by different methods.

Psychiatric disorders and Type 2 diabetes mellitus: A bidirectional Mendelian randomization.

BACKGROUND: Extensive observational evidence put forward the association between psychiatric disorders and type 2 diabetes mellitus (T2DM). However, causal relationships between these two diseases required further research. Thus, we evaluated the bidirection casual effect between five psychiatric disorders and T2DM using two-sample mendelian randomization (MR). METHODS: By selecting single nucleotide polymorphisms associated with T2DM and five psychiatric disorders (attention-deficit hyperactivity disorder (ADHD), major depressive disorder (MDD), schizophrenia, anxiety disorder and panic disorder), a bidirectional two-sample MR was applied to evaluate causality between these diseases. The inverse-variance weighted (IVW) method was used as the primary analysing approach for estimating possible causal effects. MR-Egger and weighted median were also conducted to verify the results. The funnel plot, Cochran's Q test and MR-Egger intercept test were used for sensitivity analyses. In addition, potential mediators were investigated by risk factor analyses. RESULTS: Genetic susceptibilities of ADHD and MDD would increase the risk of T2DM (ADHD: OR = 1.14, 95%CI 1.08-1.20; p = 5.7 × 10 - 6 ; MDD: OR = 1.22, 95%CI 1.09-1.36; p = 0.0004 ). In addition, genetic predisposition to T2DM was also associated with ADHD (OR = 1.09, 95%CI 1.04-1.14; p = 0.0004). Several risk factors of T2DM were implicated in the above causal associations, including smoking, high body mass index, waist-to-hip ratio and elevated serum triglycerides. CONCLUSION: Our studies indicated a causal effect of ADHD and MDD on increasing the risk of T2DM, which was potentially mediated by smoking and obesity-related phenotypes. Meanwhile, we found a causal effect of T2DM on ADHD. Thus, prevention strategies for T2DM should also include mental health and vice versa.

Research on adaptive impedance control technology of upper limb rehabilitation robot based on impedance parameter prediction.

Introduction: With the aggravation of aging and the growing number of stroke patients suffering from hemiplegia in China, rehabilitation robots have become an integral part of rehabilitation training. However, traditional rehabilitation robots cannot modify the training parameters adaptively to match the upper limbs' rehabilitation status automatically and apply them in rehabilitation training effectively, which will improve the efficacy of rehabilitation training. Methods: In this study, a two-degree-of-freedom flexible drive joint rehabilitation robot platform was built. The forgetting factor recursive least squares method (FFRLS) was utilized to estimate the impedance parameters of human upper limb end. A reward function was established to select the optimal stiffness parameters of the rehabilitation robot. Results: The results confirmed the effectiveness of the adaptive impedance control strategy. The findings of the adaptive impedance control studies showed that the adaptive impedance control had a significantly greater reward than the constant impedance control, which was in line with the simulation results of the variable impedance control. Moreover, it was observed that the levels of robot assistance could be suitably modified based on the subject's different participation. Discussion: The results facilitated stroke patients' upper limb rehabilitation by enabling the rehabilitation robot to adaptively change the impedance parameters according to the functional status of the affected limb. In clinic therapy, the proposed control strategy may help to adjust the reward function for different patients to improve the rehabilitation efficacy eventually.

Harnessing stepping-stone hosts to engineer, select, and reboot synthetic bacteriophages in one pot.

Advances in synthetic genomics have led to a great demand for genetic manipulation. Trimming any process to simplify and accelerate streamlining of genetic code into life holds great promise for synthesizing and studying organisms. Here, we develop a simple but powerful stepping-stone strategy to promote genome refactoring of viruses in one pot, validated by successful cross-genus and cross-order rebooting of 90 phages infecting 4 orders of popular pathogens. Genomic sequencing suggests that rebooting outcome is associated with gene number and DNA polymerase availability within phage genomes. We integrate recombineering, screening, and rebooting processes in one pot and demonstrate genome assembly and genome editing of phages by stepping-stone hosts in an efficient and economic manner. Under this framework, in vitro assembly, yeast-based assembly, or genetic manipulation of native hosts are not required. As additional stepping-stone hosts are being developed, this framework will open doors for synthetic phages targeting more pathogens and commensals.

An Improved EMG-Driven Neuromusculoskeletal Model for Elbow Joint Muscle Torque Estimation.

The accurate measurement of human joint torque is one of the research hotspots in the field of biomechanics. However, due to the complexity of human structure and muscle coordination in the process of movement, it is difficult to measure the torque of human joints in vivo directly. Based on the traditional elbow double-muscle musculoskeletal model, an improved elbow neuromusculoskeletal model is proposed to predict elbow muscle torque in this paper. The number of muscles in the improved model is more complete, and the geometric model is more in line with the physiological structure of the elbow. The simulation results show that the prediction results of the model are more accurate than those of the traditional double-muscle model. Compared with the elbow muscle torque simulated by OpenSim software, the Pearson correlation coefficient of the two shows a very strong correlation. One-way analysis of variance (ANOVA) showed no significant difference, indicating that the improved elbow neuromusculoskeletal model established in this paper can well predict elbow muscle torque.

Transcriptional reprogramming strategies and miRNA-mediated regulation networks of Taxus media induced into callus cells from tissues.

BACKGROUND: Taxus cells are a potential sustainable and environment-friendly source of taxol, but they have low survival ratios and slow grow rates. Despite these limitations, Taxus callus cells induced through 6 months of culture contain more taxol than their parent tissues. In this work, we utilized 6-month-old Taxus media calli to investigate their regulatory mechanisms of taxol biosynthesis by applying multiomics technologies. Our results provide insights into the adaptation strategies of T. media by transcriptional reprogramming when induced into calli from parent tissues. RESULTS: Seven out of 12 known taxol, most of flavonoid and phenylpropanoid biosynthesis genes were significantly upregulated in callus cells relative to that in the parent tissue, thus indicating that secondary metabolism is significantly strengthened. The expression of genes involved in pathways metabolizing biological materials, such as amino acids and sugars, also dramatically increased because all nutrients are supplied from the medium. The expression level of 94.1% genes involved in photosynthesis significantly decreased. These results reveal that callus cells undergo transcriptional reprogramming and transition into heterotrophs. Interestingly, common defense and immune activities, such as "plant-pathogen interaction" and salicylic acid- and jasmonic acid-signaling transduction, were repressed in calli. Thus, it's an intelligent adaption strategy to use secondary metabolites as a cost-effective defense system. MiRNA- and degradome-sequencing results showed the involvement of a precise regulatory network in the miRNA-mediated transcriptional reprogramming of calli. MiRNAs act as direct regulators to enhance the metabolism of biological substances and repress defense activities. Given that only 17 genes of secondary metabolite biosynthesis were effectively regulated, miRNAs are likely to play intermediate roles in the biosynthesis of secondary metabolites by regulating transcriptional factors (TFs), such as ERF, WRKY, and SPL. CONCLUSION: Our results suggest that increasing the biosynthesis of taxol and other secondary metabolites is an active regulatory measure of calli to adapt to heterotrophic culture, and this alteration mainly involved direct and indirect miRNA-induced transcriptional reprogramming. These results expand our understanding of the relationships among the metabolism of biological substances, the biosynthesis of secondary metabolites, and defense systems. They also provide a series of candidate miRNAs and transcription factors for taxol biosynthesis.

Knockdown of GRHL3 inhibits activities and induces cell cycle arrest and apoptosis of human colorectal cancer cells.

The Grainyhead-like 3 (GRHL3) is involved in epidermal barrier formation, neural tube closure and wound repair. Previous studies have suggested that GRHL3 has been linked to many different types of cancers. However, to date, its effects on human colorectal cancer (CRC) has not been clarified yet. Our microarray analysis has indicated predominant GRHL3 expression in CRC. The purpose of this study was to investigate the expression and significance of GRHL3 in CRC tumorigenesis using CRC tissues and paired paracancerous tissues, as well as using distinct CRC cell lines (HT29 and DLD1). We observed increased GRHL3 expression at both mRNA and protein levels in CRC tissues and CRC cell lines using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Moreover, silencing GRHL3 with siRNA could suppress CRC cell proliferation, viability and migration in vitro. We also found that knockdown of GRHL3 could promote cell cycle arrest at G0/G1 phase in HT29 cells and DLD1 cells, and induce cell apoptosis in HT29 cells. Together, our study revealed the down-regulation of GRHL3 in vitro could inhibit CRC cell activity and trigger cell cycle arrest at G0/G1 phase and apoptosis.

Inhibition of adhesion and metastasis of HepG2 hepatocellular carcinoma cells in vitro by DNA aptamer against sialyl Lewis X.

The sialyl Lewis X (SLex) antigen encoded by the FUT7 gene is the ligand of endotheliam-selectin (E-selectin). The combination of SLex antigen and E-selectin represents an important way for malignant tumor metastasis. In the present study, the effect of the SLex-binding DNA aptamer on the adhesion and metastasis of hepatocellular carcinoma HepG2 cells in vitro was investigated. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining were conducted to detect the expression of FUT7 at both transcriptional and translational levels. The SLex expression in HepG2 cells treated with different concentrations of SLex-binding DNA aptamer was detected by flow cytometry. Besides, the adhesion, migration, and invasion of HepG2 cells were measured by cell adhesion assay, and the Transwell migration and invasion assay. The results showed that the FUT7 expression was up-regulated at both mRNA and protein levels in HepG2 cells. SLex-binding DNA aptamer could significantly decrease the expression of SLex in HepG2 cells. The cell adhesion assay revealed that the SLex-binding DNA aptamer could effectively inhibit the interactions between E-selectin and SLex in the HepG2 cells. Additionally, SLex-binding DNA aptamers at 20 nmol/L were found to have the similar effect to the monoclonal antibody CSLEX-1. The Transwell migration and invasion assay revealed that the number of penetrating cells on the down-side of Transwell membrane was significantly less in cells treated with 5, 10, 20 nmol/L SLex-binding DNA aptamer than those in the negative control group (P<0.01). Our study demonstrated that the SLex-binding DNA aptamer could significantly inhibit the in vitro adhesion, migration, and invasion of HepG2 cells, suggesting that the SLex-binding DNA aptamer may be used as a potential molecular targeted drug against metastatic hepatocellular carcinoma.

Inhibition of Adhesion and Metastasis of HepG2 Hepatocellular Carcinoma Cells In Vitro by DNA Aptamer against Sialyl Lewis X / 华中科技大学学报（医学）（英德文版）

The sialyl Lewis X (SLex) antigen encoded by the FUT7 gene is the ligand of endotheliam-selectin (E-selectin).The combination of SLex antigen and E-selectin represents an important way for malignant tumor metastasis.In the present study,the effect of the SLeX-binding DNA aptamer on the adhesion and metastasis of hepatocellular carcinoma HepG2 cells in vitro was investigated.Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining were conducted to detect the expression of FUT7 at both transcriptional and translational levels.The SLex expression in HepG2 cells treated with different concentrations of SLeX-binding DNA aptamer was detected by flow cytometry.Besides,the adhesion,migration,and invasion of HepG2 cells were measured by cell adhesion assay,and the Transwell migration and invasion assay.The results showed that the FUT7 expression was up-regulated at both mRNA and protein levels in HepG2 cells.SLeX-binding DNA aptamer could significantly decrease the expression of SLex in HepG2 cells.The cell adhesion assay revealed that the SLeX-binding DNA aptamer could effectively inhibit the interactions between E-selectin and SLex in the HepG2 cells.Additionally,SLeX-binding DNA aptamers at 20 nmol/L were found to have the similar effect to the monoclonal antibody CSLEX-1.The Transwell migration and invasion assay revealed that the number of penetrating cells on the down-side of Transwell membrane was significantly less in cells treated with 5,10,20 nmol/L SLeX-binding DNA aptamer than those in the negative control group (P＜0.01).Our study demonstrated that the SLeX-binding DNA aptamer could significantly inhibit the in vitro adhesion,migration,and invasion of HepG2 cells,suggesting that the SLeX-binding DNA aptamer may be used as a potential molecular targeted drug against metastatic hepatocellular carcinoma.

Knockdown of GRHL3 Inhibits Activities and Induces Cell Cycle Arrest and Apoptosis of Human Colorectal Cancer Cells / 华中科技大学学报（医学）（英德文版）

The Grainyhead-like 3 (GRHL3) is involved in epidermal barrier formation,neural tube closure and wound repair.Previous studies have suggested that GRHL3 has been linked to many different types of cancers.However,to date,its effects on human colorectal cancer (CRC) has not been clarified yet.Our microarray analysis has indicated predominant GRHL3 expression in CRC.The purpose of this study was to investigate the expression and significance of GRHL3 in CRC tumorigenesis using CRC tissues and paired paracancerous tissues,as well as using distinct CRC cell lines (HT29 and DLD1).We observed increased GRHL3 expression at both mRNA and protein levels in CRC tissues and CRC cell lines using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting.Moreover,silencing GRHL3 with siRNA could suppress CRC cell proliferation,viability and migration in vitro.We also found that knockdown of GRHL3 could promote cell cycle arrest at G0/G1 phase in HT29 cells and DLD1 cells,and induce cell apoptosis in HT29 cells.Together,our study revealed the down-regulation of GRHL3 in vitro could inhibit CRC cell activity and trigger cell cycle arrest at G0/G1 phase and apoptosis.