Objective Improving the existed HPLC methods in order to detect the levels of global DNA methylation rapidly, stably and conveniently. Methods The HP1100 high performance liquid chrom-atographic (HPLC) system was used in this study. The analytic column was Macherey-Nagel (MN) EC 250/4.6 Nucleosil 100-5SA (250mm x 4. 6mm, 5u,m) cation exchange chromatographic column. We used 60mM acetic acid + 15% acetonitrile as mobile phase (adjust to pH =4. 6 by NaOH). The flow rate was 1. 0 ml/min, detective UV wavelength was set to 276 nm and column temperature was set to 28℃. The injection volume was 50μl. The global DNA methylation was expressed as 5mdC/(dC +5mdC) x 100%. Results Under these conditions, we can isolate dC and 5mdC completely in ten minutes. The level of leukocyte global DNA methylation in healthy people is (4. 389 ±0. 0159) %. Conclusions This method can determine the levels of global DNA methylation rapidly, and it can be widely applied in some laboratories.

Objective To compare the real-time dissolutions of 3 kinds of Simvastatin tablets used in our hospital, and provide reference information for clinical applications. Methods The dissolution rate of simvastatin tablets was deter-mined by the Chinese Pharmacopoeia 2010, and the HPLC method was used to determine the simvastatin content at the wavelength of 238 nm and the Weibull’s equation was adopted to fit the dissolution curves. The shape parameters (m) were performed statistical analysis; Using the similarity f2 as the index, compared the dissolution curve between tablets and the preparation. Results The dissolution of simvastatin tablets manufactured by the 3 manufaccturer was no less than 90% in 30 min, which comply with the national specifications. Compared with the dissolution curve of 6 tablets from the same batch, the homogenicity of samples from A was better, with poor homogenicity of tablets from B or C. There’s significant difference in the shape parameter m of tablets from B and C (P< 0.05), and there’s no significant difference in m of tablets from C and A(P>0.05). Conclusion The in vitro dissolution of the simvastatin tablets are de-termined with single point test procedures. The dissolution of the tablets from 3 manufactures comply with national specifications. However, there’s significant difference in real time dissolution of national tablets. It’s necessary to im-prove the quality of national tablets. Cautions should be taken while use in clinical practices.

The basic information and clinical application of nutritional risk scales for children with cancer were reviewed, and the strengths and weaknesses of each scale were analyzed. After systematic search and reading, the scales with more clinical applications included universal scales: Pediatric Malnutrition Assessment Screening Tool (STAMP), Nutritional Risk and Stunting Malnutrition Screening Tool (STRONG kids), Pediatric Yorkhill Malnutrition Score (PYMS), Pediatric Subjective Global Nutritional Risk Assessment (SGNA); specific scales: Nutritional Screening Tool for Childhood Cancer (SCAN), Nutritional Risk Screening for Childhood Cancer (NRS-PC). In order to effectively manage the nutritional risk of pediatric cancer patients, we should selectively use and further actively Chinese or develop specific nutritional risk measurement tools adapted to our national conditions.

OBJECTIVE To evaluate the efficacy and safety of Huoxue Xiaoyi Granule in inhibiting the recurrence of endometrio-sis(EMs)after conservative operation,and to provide evidence for Chinese medicine in inhibiting the recurrence of EMs.METHODS A total of 72 patients with qi-stagnation and blood-stasis after EMs operation were selected as the study objects and randomly divid-ed into the Chinese medicine group and the Western medicine group,36 cases in each group.The Chinese medicine group was treated with Huoxue Xiaoyi Granule,and the Western medicine group was treated with GnRH-α.The postoperative recurrence rate,TCM syn-drome score,carbohydrate antigen 125(CA125),dysmenorrhea score,pelvic pain score,pregnancy rate,serum sex hormones[estra-diol(E2),luteinizing hormone(LH),follicle stimulating hormone(FSH)]and safety indexes of the two groups were observed.RE-SULTS At 9 months and 12 months after surgery,the total TCM syndrome scores of the two groups of patients were significantly re-duced(P<0.05),and the Chinese medicine group was better than the Western medicine group(P<0.05);12 months later,the TCM total clinical curative rate in the Chinese medicine group was better than that in the Western medicine group;after surgery,the serum CA125 levels,dysmenorrhea scores and pelvic pain severity of the two groups of patients were significantly reduced(P<0.05,P<0.01);during treatment,the total incidence of adverse reactions in the Chinese medicine group was lower than that in the Western medicine group(P<0.01);the recurrence rate of the Chinese medicine group was slightly lower than that of the Western medicine group,but there was no statistical difference(P>0.05).CONCLUSION Huoxue Xiaoyi Granule can significantly improve TCM syndromes in patients with qi-stagnation and blood-stasis after EMs surgery.It is safe and has equivalent efficacy to GnRH-α in pre-venting postoperative recurrence of EMs,and worthy of clinical promotion and application.

Objective:To compare the genotypes and phenotypes between the monozygotic twins via whole genome sequencing to further clarify the autosomal dominant inherited neurofibromatosis type 1 (NF1) variants related to congenital pseudarthrosis (CP).Methods:According to the diagnostic criteria of congenital tibial pseudarthrosis and the clinical diagnostic criteria of NF1, two pairs of monozygotic twins with NF1 were included. Both were female and only one of each pair had congenital pseudarthrosis. The other did not have congenital pseudarthrosis. Whole genome sequencing was performed using the peripheral blood of the two pairs of monozygotic twins. Customized bioinformatics analysis was then performed to identify single nucleotide variants (SNVs), short insertion deletion variants (InDel), copy number variants (CNVs), and structural variants (SVs). Classified the variants according to the American College of Medical Genetics and Genomics (ACMG) and ClinGen criteria. The germline variants within the monozygotic twins were compared to identify the CP patients' unique variants. The shared pathogenic or likely pathogenic germline variants between the unique variants in the CP patients from the twins were also analyzed. Further, the identified disease-causing variants were validated by Sanger sequencing in the family of the twins and their parents. Finally, the genotypes and phenotypes regarding the pathogenic variants of the NF1 gene among the twins were characterized. Results:Both the two monozygotic twins were identified pathogenic variants in the NF1 gene. One with c.3047_3048del (p.Cys1016SerfsTer4), and the other with c.4267A>G (p.Lys1423Glu). By Sanger sequencing validation in family quads, the two CP patients and their siblings harbored de novo heterozygous variants of the NF1 gene. In addition to the NF1 gene, no other genes were identified pathogenic or likely pathogenic variants uniquely in the CP patients compared with their twin sisters, as well as SVs and CNVs. In addition, by analyzing the rare and damaging variants in the two CP patients from the two twins, they had no overlapping genes against the SNVs, InDels, SVs, or CNVs. Conclusion:Whole genome sequencing revealed that both the two monozygotic twins with NF1 were detected pathogenic variants of gene NF1. No other pathogenic variants specific to the CP patients among the twins were identified. The two CP patients shared no other common genes from the detected likely pathogenic variants.

Congenital joint synostosis (CJS) is a functional impairment resulting from failure in joint morphogenesis during embryonic development. Clinically, it may be classified as syndromic (sCJS) and non-syndromic (nsCJS) disorders. Common sCJS include chromosomal disorders such as Klinefelter syndrome and single-gene disorders like Apert/Pfeiffer/Crouzon syndromes, Holt-Oram syndrome, Ehlers-Danlos syndrome, and Radial-ulnar synostosis with thrombocytopenia, presenting with multiple system/organ anomalies. By contrast, nsCJS manifest with only joint abnormalities, affecting one or multiple joints. This review has focused on human nsCJS and its genetic etiology. To date, variants in seven genes ( NOG, GDF5, FGF9, GDF6, FGF16, SMAD6, and MECOM) have been identified as causative factors for nsCJS. This review has focused on such genes and provided a comprehensive review for the clinical phenotypes, genetic patterns, common variants, and underlying mechanisms associated with nsCJS based on a literature review. In addition, it has also analyzed other candidate genes for nsCJS within the context of relevant signaling pathways involved in joint morphogenesis.

Objective To compare the hemodynamic characteristics in internal carotid artery models, which were obtained by multi-scale unidirectional and bidirectional coupling models, so as to provide references for selecting models in further studies. Methods Based on the nuclear magnetic resonance image of one patient with mild stenosis of internal carotid artery, the lumped parameter model of the circle of Willis and the three-dimensional model of internal carotid artery were constructed. Those two different multi-scale models were constructed by unidirectional and bidirectional coupling. Results With the increase of stenosis degree, the inlet and outlet blood pressure and the outlet blood flow of internal carotid artery all decreased under two kinds of coupling method. The distribution of low time average wall shear stress (TAWSS) and high oscillatory shear index (OSI) of the internal carotid artery both increased with the increase of stenosis degree under two kinds of coupling method in general. The anterior cerebral artery segment showed lower shear stress and higher OSI with bidirectional coupling in 70% stenosis, and the blood flow direction of posterior communicating artery was changed, which was significantly different from unidirectional coupling results. Conclusions At a low degree of stenosis, the result of those two kinds of coupling method were consistent in general, but there was a significant difference in 70% stenosis, and the result of bidirectional coupling was closer to physiological parameters. The research findings can be better applied to the hemodynamic study of cerebrovascular diseases.

Integrating evidence from animal experiments is a critical component of biomedical research, providing essential prior information for in-depth investigations of disease mechanisms and new drug development. Animal models have played an irreplaceable role in simulating human diseases. However, the integration of evidence from animal experiments has faced numerous challenges, including insufficient emphasis, significant heterogeneity in study designs, high publication bias, and discrepancies with clinical research practices. This paper first identifies existing issues in the original research evidence from animal experiments, such as the selection and applicability of animal models, considerations in the design of experimental studies, and factors influencing the translation of animal experimental evidence. It then discusses various methods for integrating this evidence, including systematic review and meta-analysis, overview of systematic review/umbrella review, scoping review, and evidence mapping, while highlighting recent advancements in their application. Finally, the paper addresses the main challenges currently encountered in the integration of evidence from animal experiments and proposes targeted improvement strategies aimed at enhancing the efficiency of translating research outcomes into clinical practice and promoting the advancement of evidence-based medicine. By continuously optimizing original experimental research protocols and evidence integration practices, this work aims to establish a more efficient and scientific environment for the synthesis of evidence from animal experiments, ultimately contributing to clinical trials and human health.

Objective: To investigate the ameliorative effects of Mushroom on adipose tissue inflammation and glucolipid metabolism in mice fed a high-fat diet, and to provide a theoretical basis for the mechanisms of Mushroom regulating glucolipid metabolism and inflammatory responses. Methods: C57 BL/6 J mice were fed with normal diet(LF) group, high-fat diet(HF)group and high-fat diet + Mushroom(HF+Mushroom) group for 15 weeks.Then, body weight subcutaneous and epididymal white adipose tissue weight were measured. The morphological changes of adipose tissues were compared by HE staining, and the expression of genes related to inflamation, glycolysis and fatty acid oxidation pathways were detected by RT-PCR and Western blot. Results: Compared with the LF group, the HF group had increased body weight, increased subcutaneous and epididymal white fat weight and adipocyte size, and upregulated expression of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), monocyte chemoattractant protein-1(MCP-1), CD68, inducible nitric oxide synthase(iNOS), pyruvate kinase(PK), phosphofructokinase(PFK), hypoxia inducible factor-1α(HIF-1α) and peroxisome proliferator activated receptor alpha(PPARα) in adipose tissues, while the expression of carnitine palmitoyl transferase-1 A(CPT-1 A), cytochrome P450 4 a10(CYP4 a10) and medium-chain acyl-coenzyme a dehydrogenase(MCAD) were downregulated(P<0.05). Compared with the HF group, Mushroom supplementation reduced body weight, adipose tissue weight and adipocyte size, and downregulated the expression of pro-inflammatory factors and glycolytic pathway-related factors in adipose tissues, while the expression of fatty acid oxidation pathway-related factors were upregulated(P<0.05). Conclusion Mushroom can ameliorate inflammation and disorders of glycolipid metabolism in adipose tissues of obese mice.