Objective This study aimed to understand the epidemic status and phylogenetic relationships of rotavirus group A(RVA)in the Pearl River Delta region of Guangdong Province,China. Methods This study included individuals aged 28 days-85 years.A total of 706 stool samples from patients with acute gastroenteritis collected between January 2019 and January 2020 were analyzed for 17 causative pathogens,including RVA,using a Gastrointestinal Pathogen Panel,followed by genotyping,virus isolation,and complete sequencing to assess the genetic diversity of RVA. Results The overall RVA infection rate was 14.59%(103/706),with an irregular epidemiological pattern.The proportion of co-infection with RVA and other pathogens was 39.81%(41/103).Acute gastroenteritis is highly prevalent in young children aged 0-1 year,and RVA is the key pathogen circulating in patients 6-10 months of age with diarrhea.G9P[8](58.25%,60/103)was found to be the predominant genotype in the RVA strains,and the 41 RVA-positive strains that were successfully sequenced belonged to three different RVA genotypes in the phylogenetic analysis.Recombination analysis showed that gene reassortment events,selection pressure,codon usage bias,gene polymorphism,and post-translational modifications(PTMs)occurred in the G9P[8]and G3P[8]strains. Conclusion This study provides molecular evidence of RVA prevalence in the Pearl River Delta region of China,further enriching the existing information on its genetics and evolutionary characteristics and suggesting the emergence of genetic diversity.Strengthening the surveillance of genotypic changes and gene reassortment in RVA strains is essential for further research and a better understanding of strain variations for further vaccine development.

Based on the genomic information of Emericella sp. 1454, in conjunction with literature analysis of its secondary metabolite emestrin, this study identified the biosynthetic precursors of emestrin and enhanced its production by supplementing the culture medium with these precursors. In this study, it was found for the first time that the addition of biosynthetic precursor, reduced glutathione, to the culture medium significantly increased emestrin yield. By incorporating 1.5 g of reduced glutathione into 50 g of rice culture medium and fermenting for 15 days, a yield of 30.82 mg of emestrin was obtained, which marked an 11.71-fold increase compared to the original fermentation approach. The method is both simple and cost-effective, establishing a solid foundation for the efficient synthesis of emestrin and similar compounds. Additionally, it serves as an important reference for enhancing the production of other epipolythiodioxopiperazine compounds.

mRNA gene therapy has attracted much attention due to its advantages such as scalability, modification, no need to enter the nucleus and no integration of host genes. In gene therapy, safe and effective delivery of mRNA into cells is critical for the success of gene therapy. In this study, we designed and synthesized an amphiphilic cationic lipopeptide gene vector (dendritic arginine & disulfide bond-containing cationic lipopeptide, RLS) enriched with branched arginine. We achieved a 1.5-fold higher mRNA transfection efficiency in zebrafish compared to the commercial reagent Lipofectamine 2000, and confirmed its good biosafety by in vitro cytotoxicity and in vivo biosafety. First, we characterized the chemical composition of the cationic lipid peptides by nuclear magnetic resonance hydrogen spectroscopy (¹H NMR) and time-of-flight mass spectrometry (MS). The results of particle size and potential tested by dynamic light scattering particle size analysis showed that at a nitrogen/phosphorus (N/P) ratio of 20, the RLS/mRNA composite assemblies formed homogeneous nanoparticles with an average particle size of about 220 nm and a surface ζ potential of about +21 mV. In vitro gene transfection, the transfection experiments demonstrated that RLS exhibited 1.2-fold higher transfection efficiency in human embryonic kidney 293 cells (HEK293) and 3-fold higher transfection efficiency in rat mesenchymal stem cells (MSC) compared to Lipofectamine 2000. In addition, after microinjection of RLS into zebrafish embryos, we evaluated the survival, hatching, and teratogenicity rates, all of which confirmed its favorable in vivo safety profile. Thus, this amphiphilic cationic lipid peptide RLS, enriched with branched arginine, exhibits excellent mRNA delivery properties and safety. These findings highlight its potential as a promising gene therapy tool.

AIM To investigate the effects of total Astragalus saponins on the improvement of sarcopenia in a rat model of type 2 diabetes mellitus(T2DM).METHODS The rats were divided into the normal group for a normal feeding and the model group for the feeding of high-sugar and high-fat diet combined with intraperitoneal injection of STZ to establish a T2DM model.The successful model rats were randomly divided into the model group,the metformin group(0.2 g/kg)and the total Astragalus saponins group(80 mg/kg),and given corresponding doses of drugs by gavage.After 12 weeks administration,the rats had their FBG,postprandial blood glucose(PG2h)and wet weight of skeletal muscle measured;their serum levels of INS,C-peptide(C-P),IGF-1,TNF-α and IL-1β detected by ELISA;their morphological changes of skeletal muscle observed by HE staining;their protein expressions of PI3K,p-Akt,mTOR,S6K1,FoxO1 and Murf1 in skeletal muscle detected by Western blot;and their mRNA expressions of Pi3k,Akt and mtor in skeletal muscle detected by RT-qPCR method.RESULTS Compared with the model group,the total Astragalus saponins group displayed decreased levels of FBG,PG2h,OGTT-AUC,HOMA-IR,TNF-α and IL-1β(P＜0.01);increased levels of INS,C-P,IGF-1 and wet weight of skeletal muscle(P＜0.05,P＜0.01);improved skeletal muscle atrophy and increased protein expressions of PI3K,p-Akt,mTOR and S6K1 in skeletal muscle(P＜0.05,P＜0.01);decreased protein expressions of FoxO1 and Murf1(P＜0.05,P＜0.01);and increased mRNA expressions of Pi3k,Akt and mtor(P＜0.01).CONCLUSION The improvement effects of total Astragalus saponins on sarcopenia in T2DM rats may be associated with the regulation of PI3K/Akt/mTOR and PI3K/Akt/FoxO1 pathways.

Ovarian cancer is one of the most fatal of all the gynecological malignancies.The detection of biomarker is one of the diagnostic bases for the disease.The combined detection of carbohydrate antigen 125(CA125)and human epididymis protein 4(HE4)in human serum can improve the accuracy of clinical diagnosis for ovarian cancer.In this study,a new method for analysis of ovarian cancer biomarkers CA125 and HE4 in human serum using zinc oxide quantum dot(ZnO QD)and tin oxide quantum dot(SnO2 QD)labeled antibodies coupled with single particle inductively coupled plasma mass spectrometry(ICP-MS)was developed.CA125 and HE4 antigens were captured by amino-modified magnetic nanoparticles combined with CA125 and HE4 primary antibodies.CA125 and HE4 secondary antibodies were labeled by ZnO QD and SnO2 QD,respectively.The signal frequency of Zn and Sn was determined by ICP-MS to realize the quantitative analysis of CA125 and HE4.The linear detection ranges of CA125 and HE4 were 0.02-200 U/mL and 0.02-100 ng/mL,the detection limits were 0.004 U/mL and 0.006 ng/mL(3σ),and the RSDs were 2.2％and 3.5％(n=6),respectively.This method had many advantages such as high sensitivity,good selectivity and less reagent consumption,and was successfully applied to determine the content of CA125 and HE4 in human serum.

Objective To evaluate the short-term efficacy of Huoxue Xiaozhong Powder(i.e.,Shuangbai San)for the treatment of patients with mild-to-moderate knee osteoarthritis(KOA)of qi stagnation and blood stasis syndrome.Methods Sixty patients with mild-to-moderate KOA of qi stagnation and blood stasis syndrome were randomly divided into the treatment group and the control group,with 30 cases in each group.The treatment group was treated with external application of Huoxue Xiaozhong Powder,and the control group was treated with external application of Voltaren(diclofenac diethylamide emulsion).Both groups were treated for one week as a course.The changes of visual analogue scale(VAS)score,Western Ontario and McMaster Universities Osteoarthritis Index(WOMAC)score and erythrocyte sedimentation rate(ESR),C-reactive protein(CRP)level and synovial thickness under ultrasound in the two groups before and after treatment were observed.Moreover,the clinical efficacy and safety of the two groups were evaluated.Results(1)After one week of treatment,the total effective rate of the treatment group was 93.33%(28/30),and that of the control group was 73.33%(22/30).The intergroup comparison(tested by rank sum test)showed that the clinical efficacy of the treatment group was significantly superior to that of the control group(P＜0.05).(2)After treatment,the VAS score,WOMAC score,ESR,CRP level and synovial thickness under ultrasound in the two groups were significantly improved compared with those before treatment(P＜0.01),and the treatment group had stronger effect on the improvement of VAS score,WOMAC score,ESR,CRP level and synovial thickness under ultrasound than the control group(P＜0.01).(3)During treatment,no obvious adverse reactions such as skin redness,swelling and itching occurred in the two groups of patients,which had a high safety.Conclusion For the patients with mild-to-moderate KOA of qi stagnation and blood stasis syndrome,the use of Huoxue Xiaozhong Powder for external treatment alone can also achieve good short-term clinical effects,which can effectively alleviate the symptoms of pain,and reduce the levels of inflammatory indicators and synovial thickness.

Bone defects caused by different causes such as trauma, severe bone infection and other factors are common in clinic and difficult to treat. Usually, bone substitutes are required for repair. Current bone grafting materials used clinically include autologous bones, allogeneic bones, xenografts, and synthetic materials, etc. Other than autologous bones, the major hurdles of rest bone grafts have various degrees of poor biological activity and lack of active ingredients to provide osteogenic impetus. Bone marrow contains various components such as stem cells and bioactive factors, which are contributive to osteogenesis. In response, the technique of bone marrow enrichment, based on the efficient utilization of components within bone marrow, has been risen, aiming to extract osteogenic cells and factors from bone marrow of patients and incorporate them into 3D scaffolds for fabricating bone grafts with high osteoinductivity. However, the scientific guidance and application specification are lacked with regard to the clinical scope, approach, safety and effectiveness. In this context, under the organization of Chinese Orthopedic Association, the Expert consensus for the clinical application of autologous bone marrow enrichment technique for bone repair ( version 2023) is formulated based on the evidence-based medicine. The consensus covers the topics of the characteristics, range of application, safety and application notes of the technique of autologous bone marrow enrichment and proposes corresponding recommendations, hoping to provide better guidance for clinical practice of the technique.

Objective:To analyze the risk factors of acute kidney injury (AKI) in hip fracture patients with serious underlying diseases and establish a prediction nomogram.Methods:Clinical information of hip fracture patients admitted to the intensive care unit (ICU) of Beth Israel Deaconess Medical Center (BIDMC) was analyzed using the Medical Information Mart for Intensive Care (MIMIC)-IV. Patient comorbidities, disease scores, vital signs and laboratory tests, surgical modalities, invasive procedures, and drug use were recorded. According to the diagnostic criteria of AKI in the Kidney Disease Improving Global Outcome (KDIGO) guideline, the enrolled patients were randomly divided into training set and validation set. Based on logistic regression analysis, least absolute shrinkage and selection operator (LASSO) logistic regression algorithm was used to analyze the risk factors of AKI after admission, and the corresponding prediction model was calculated.Results:A total of 474 patients were enrolled, including 331 in the training set and 143 in the validation set. According to the diagnostic criteria of AKI of KDIGO guidelines, the patients were divided into AKI group (159 cases) and non-AKI group (172 cases). Univariate analysis showed that age ( t=2.61, P=0.009), coronary heart disease (χ 2=2.08, P=0.038), heart failure (χ 2=2.60, P=0.009), hemoglobin ( t=1.89, P=0.059), platelets ( t=1.81, P=0.070), urea nitrogen ( t=2.83, P=0.005), blood creatinine ( t=3.65, P<0.001), blood sodium ( t=2.55, P=0.011), blood glucose ( t=2.52, P=0.012), anion gap ( t=3.44, P=0.001), diastolic blood pressure ( t=2.72, P=0.007), mean arterial pressure ( t=2.16, P=0.031), SOFA score ( t=3.69, P<0.001), simplified acute physiological function score II (SAPSII) score ( t=2.95, P=0.003), as well as furosemide (χ 2=2.03, P=0.042), vancomycin (χ 2=1.70, P=0.089), vasoactive medications (χ 2=3.74, P<0.001) and use of invasive mechanical ventilation (χ 2=4.81, P<0.001) were risk factors associated with the development of AKI in hip fracture patients. Multivariate logistic regression analysis showed that age ( OR=1.03, P<0.001), coronary heart disease ( OR=2.05, P=0.069), hemoglobin ( OR=0.88, P=0.050), blood creatinine ( OR=1.37, P=0.009), blood sodium ( OR=1.07, P=0.026), anion gap ( OR=1.09, P=0.028) and vasoactive medications ( OR=3.83, P=0.018) and the use of invasive mechanical ventilation ( OR=6.56, P<0.001) were independent predictors of the development of AKI in hip fracture patients with serious underlying diseases. The area under the curve of the nomogram prediction model constructed by the above 8 predictors was 0.789, and the calibration curve of the nomogram was close to the ideal diagonal. Decision curve analysis showed that the net benefit of the model was significant. Conclusion:The incidence of AKI is high in hip fracture patients with serious underlying diseases. Age, coronary heart disease, hemoglobin, serum creatinine, serum sodium, anion gap, vasoactive drugs, and invasive mechanical ventilation can predict the occurrence of AKI to a certain extent. Combined with the risk factors, the construction of the corresponding prediction model can predict and manage the diagnosis and treatment of AKI in patients with hip fracture complicated with severe underlying diseases.

Objective: Glycogen storage disease type IX (GSD-IX) is a rare primary glucose metabolism abnormality caused by phosphorylase kinase deficiency and a series of pathogenic gene mutations. The clinical characteristics, gene analysis, and functional verification of a mutation in a child with hepatomegaly are summarized here to clarify the pathogenic cause of the disease. Methods: The clinical data of a child with GSD-IX was collected. Peripheral blood from the child and his parents was collected for genomic DNA extraction. The patient's gene diagnosis was performed by second-generation sequencing. The suspected mutations were verified by Sanger sequencing and bioinformatics analysis. The suspected splicing mutations were verified in vivo by RT-PCR and first-generation sequencing. Results: Hepatomegaly, transaminitis, and hypertriglyceridemia were present in children. Liver biopsy pathological examination results indicated glycogen storage disease. Gene sequencing revealed that the child had a c.285 + 2_285 + 5delTAGG hemizygous mutation in the PHKA2 gene. Sanger sequencing verification showed that the mother of the child was heterozygous and the father of the child was of the wild type. Software such as HSF3.1 and ESEfinder predicted that the gene mutation affected splicing. RT-PCR of peripheral blood from children and his mother confirmed that the mutation had caused the skipping of exon 3 during the constitutive splicing of the PHKA2 gene. Conclusion: The hemizygous mutation in the PHKA2 gene (c.285 + 2_285 + 5delTAGG) is the pathogenic cause of the patient's disease. The detection of the novel mutation site enriches the mutation spectrum of the PHKA2 gene and serves as a basis for the family's genetic counseling.
Child ; Humans ; Exons ; Glycogen Storage Disease/genetics* ; Hepatomegaly/genetics* ; Mutation ; Phosphorylase Kinase/genetics* ; Male ; Female

Background: and Purpose We investigated the causal relationships between the gut microbiota (GM), stroke, and potential metabolite mediators using Mendelian randomization (MR). Methods: We leveraged the summary statistics of GM (n=18,340 in the MiBioGen consortium), blood metabolites (n=115,078 in the UK Biobank), and stroke (cases n=60,176 and controls n=1,310,725 in the Global Biobank Meta-Analysis Initiative) from the largest genome-wide association studies to date. We performed bidirectional MR analyses to explore the causal relationships between the GM and stroke, and two mediation analyses, two-step MR and multivariable MR, to discover potential mediating metabolites. Results: Ten taxa were causally associated with stroke, and stroke led to changes in 27 taxa. In the two-step MR, Bifidobacteriales order, Bifidobacteriaceae family, Desulfovibrio genus, apolipoprotein A1 (ApoA1), phospholipids in high-density lipoprotein (HDL_PL), and the ratio of apolipoprotein B to ApoA1 (ApoB/ApoA1) were causally associated with stroke (all P<0.044). The causal associations between Bifidobacteriales order, Bifidobacteriaceae family and stroke were validated using the weighted median method in an independent cohort. The three GM taxa were all positively associated with ApoA1 and HDL_PL, whereas Desulfovibrio genus was negatively associated with ApoB/ApoA1 (all P<0.010). Additionally, the causal associations between the three GM taxa and ApoA1 remained significant after correcting for the false discovery rate (all q-values <0.027). Multivariable MR showed that the associations between Bifidobacteriales order, Bifidobacteriaceae family and stroke were mediated by ApoA1 and HDL_PL, each accounting for 6.5% (P=0.028) and 4.6% (P=0.033); the association between Desulfovibrio genus and stroke was mediated by ApoA1, HDL_PL, and ApoB/ApoA1, with mediated proportions of 7.6% (P=0.019), 4.2% (P=0.035), and 9.1% (P=0.013), respectively. Conclusion The current MR study provides evidence supporting the causal relationships between several specific GM taxa and stroke and potential mediating metabolites.