The bioactivity-guided isolation of potentially active natural products has been widely utilized in pharmaceutical discovery. In this study, by screening fungal extracts against coxsackievirus B3 (CVB3), three new aspochalasins, templichalasins A‒C (1‒3), along with six known aspochalasins (4‒9) were isolated from an active extract derived from the endophytic fungus Aspergillus templicola LHWf045. Compound 1 features a unique 5/6/5/7/5 pentacyclic ring system, while compounds 2 and 3 possess unusual 5/6/6/7 tetracyclic skeletons. Their structures were characterized through extensive spectroscopic analyses, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction analysis. Additionally, we demonstrated that compound 4 can be readily converted into compounds 1‒3 under mild acidic conditions and proposed a plausible mechanism for this conversion. Bioactivity evaluation of compounds 1‒9 against CVB3 revealed the inhibitory effects of all compounds against the virus. Notably, compound 9 exhibited superior antiviral activity, surpassing the commercial drug ribavirin in selectivity index (SI) value.
Antiviral Agents/isolation & purification* ; Aspergillus/chemistry* ; Molecular Structure ; Enterovirus B, Human/drug effects* ; Endophytes/chemistry* ; Cytochalasins/isolation & purification* ; Drug Discovery ; Humans

Objective To investigate the effects of organophosphate flame retardant tri(2-chloroethyl)phosphate(TCPP)on reproductive function in male mice by the approach of non-targeted metabolomics.Methods A total of twelve 6-week-old SPF male CD-1 mice were randomly divided into control group and TCPP group,with 6 mice in each group.TCPP group was given TCPP(at the dose of 100 mg/kg/d)intragastrically,and control group was intragastrically adminis-trated with the same amount of corn oil(solvent control).After 6 weeks,the mice were killed,and the sperm were isolated from the epididymis.The sperm concentration and viability were analyzed.Testicular tissue sections were stained with he-matoxylin,and ki67 expression was detected by immunohistochemistry.Non-targeted metabolomics was used to detect the difference of metabolites in testicular tissue between the two groups,and to analyze the difference of metabolites and related pathway changes between the two groups.Results Compared with control group,the total sperm motility of mice in TCPP treatment group was significantly decreased(P＜0.05).HE staining showed irregular arrangement of spermatogenic tubule supporting cell layer in TCPP treatment group.The expression level of ki67 in testicular tissue of mice treated with TCPP was significantly decreased(P＜0.05).Non-targeted metabolomics detected 266 up-regulated metabolites with statistical difference.And 554 down-regulated metabolites with statistical difference,among which the largest difference multiples were organic acids and amino acid metabolites.The pathways with the highest concentration of differential metabolites in-cluded purine metabolism,nucleotide metabolism,amino acid metabolism,cofactor synthesis,etc.,which were mainly re-lated to basic cell life activities,pathophysiology and chemical carcinogenesis.Conclusion TCPP can significantly re-duce sperm motility and ki67 expression in mice at the dosage of 100 mg/kg/d,which might be related to its influence on key metabolic pathways such as purine,amino acid and pyruvate.

Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder in women of childbearing age, which can cause metabolic disorders, cardiovascular disease, ovarian cancer, uterine cancer and other complications, seriously endangering the health of the body. China has become one of the countries with the fastest increasing prevalence of PCOS, but its complex pathogenesis leads to highly heterogeneous clinical manifestations, making it difficult to completely cure. Therefore, clarifying the potential pathogenesis of PCOS is of great significance for early clinical screening, diagnosis, treatment, and prognosis. Recent studies have shown that long noncoding RNA (lncRNA) plays a dual role in the pathogenesis of PCOS and is a potential novel biomarker and intervention target. The characteristics of multi-component, multi-target, and multi-pathway action in Traditional Chinese Medicine (TCM) are consistent with the biological properties of lncRNA, which have diverse types, dual roles, and diverse locations. However, research on lncRNA mediated PCOS and how TCM can improve PCOS by regulating lncRNA is relatively scattered, which is not conducive to the recognition of its clinical value. Therefore, this article provides a systematic review of the dual regulatory mechanism, clinical value, and TCM intervention research of lncRNA in the occurrence and development of PCOS, aiming to clarify how lncRNA affects the occurrence and development of PCOS and potential treatment strategies, in order to provide new ideas for the clinical prevention and treatment of PCOS.

Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder in women of childbearing age, which can cause metabolic disorders, cardiovascular disease, ovarian cancer, uterine cancer and other complications, seriously endangering the health of the body. China has become one of the countries with the fastest increasing prevalence of PCOS, but its complex pathogenesis leads to highly heterogeneous clinical manifestations, making it difficult to completely cure. Therefore, clarifying the potential pathogenesis of PCOS is of great significance for early clinical screening, diagnosis, treatment, and prognosis. Recent studies have shown that long noncoding RNA (lncRNA) plays a dual role in the pathogenesis of PCOS and is a potential novel biomarker and intervention target. The characteristics of multi-component, multi-target, and multi-pathway action in Traditional Chinese Medicine (TCM) are consistent with the biological properties of lncRNA, which have diverse types, dual roles, and diverse locations. However, research on lncRNA mediated PCOS and how TCM can improve PCOS by regulating lncRNA is relatively scattered, which is not conducive to the recognition of its clinical value. Therefore, this article provides a systematic review of the dual regulatory mechanism, clinical value, and TCM intervention research of lncRNA in the occurrence and development of PCOS, aiming to clarify how lncRNA affects the occurrence and development of PCOS and potential treatment strategies, in order to provide new ideas for the clinical prevention and treatment of PCOS.

Objective To investigate the effects of organophosphate flame retardant tri(2-chloroethyl)phosphate(TCPP)on reproductive function in male mice by the approach of non-targeted metabolomics.Methods A total of twelve 6-week-old SPF male CD-1 mice were randomly divided into control group and TCPP group,with 6 mice in each group.TCPP group was given TCPP(at the dose of 100 mg/kg/d)intragastrically,and control group was intragastrically adminis-trated with the same amount of corn oil(solvent control).After 6 weeks,the mice were killed,and the sperm were isolated from the epididymis.The sperm concentration and viability were analyzed.Testicular tissue sections were stained with he-matoxylin,and ki67 expression was detected by immunohistochemistry.Non-targeted metabolomics was used to detect the difference of metabolites in testicular tissue between the two groups,and to analyze the difference of metabolites and related pathway changes between the two groups.Results Compared with control group,the total sperm motility of mice in TCPP treatment group was significantly decreased(P＜0.05).HE staining showed irregular arrangement of spermatogenic tubule supporting cell layer in TCPP treatment group.The expression level of ki67 in testicular tissue of mice treated with TCPP was significantly decreased(P＜0.05).Non-targeted metabolomics detected 266 up-regulated metabolites with statistical difference.And 554 down-regulated metabolites with statistical difference,among which the largest difference multiples were organic acids and amino acid metabolites.The pathways with the highest concentration of differential metabolites in-cluded purine metabolism,nucleotide metabolism,amino acid metabolism,cofactor synthesis,etc.,which were mainly re-lated to basic cell life activities,pathophysiology and chemical carcinogenesis.Conclusion TCPP can significantly re-duce sperm motility and ki67 expression in mice at the dosage of 100 mg/kg/d,which might be related to its influence on key metabolic pathways such as purine,amino acid and pyruvate.

Objective:To explore the genetic basis of a child with mental retardation and developmental delay.Methods:A child who had attended the genetic clinic of Linyi People′s Hospital from October 2023 to April 2024 was selected as the study subject. Intelligence and development were assessed with simplified Peabody scale. Electroencephalogram and imaging data were collected. Peripheral blood samples of the child and her parents were collected for the screening of genetic metabolic diseases, chromosomal karyotyping, and trio-whole genome sequencing (trio-WGS) analysis. Candidate variant was verified by Sanger sequencing, and RNAseq was carried out to verify the alternative splicing due to the candidate variant. This study has been approved by the Medical Ethics Committee of Linyi People′s Hospital (No. YX200083).Results:The patient was an 8-year-and-11-month-old girl. Both of her parents had normal phenotypes. The child was assessed by the simplified Peabody scale as having intellectual disability and developmental delay. MRI showed no definite abnormal signals within the brain parenchyma, and electroencephalogram was normal. Screening of genetic metabolic diseases showed no obvious abnormality. Chromosomal karyotype was normal. Trio-WGS has detected a c. 697+ 1G>A variant in the intron 4 of the NFIX gene, along with 9 other variants within eight genes. The c. 697+ 1G>A variant may cause abnormal splicing of the NFIX gene transcript. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c. 697+ 1G>A variant was predicted to be pathogenic (PVS1+ PS2+ PM2_Supporting), while the evidence for pathogenicity of the other 9 variants was insufficient. Conclusion:The novel de novo c. 697+ 1G>A variant of the NFIX gene probably underlay the pathogenesis of the child, which may have caused the disease by leading to abnormal splicing.

Pouchitis is a common complication of ileal pouch anal anastomosis (IPAA) in patients with ulcerative colitis or familial adenomatous polyposis and the mechanism is unknown. The dietary factors including dietary ingredients and mode are related to the occurrence and progression of pouchitis. Dietary factors may play a potential role in changing gut microbiome and regulating immune response. Therefore, adjusting the diet can prevent and treat pouchitis. This article reviews the research progress of the influence of dietary factors on pouchitis.

Pouchitis is a common complication of ileal pouch anal anastomosis (IPAA) in patients with ulcerative colitis or familial adenomatous polyposis and the mechanism is unknown. The dietary factors including dietary ingredients and mode are related to the occurrence and progression of pouchitis. Dietary factors may play a potential role in changing gut microbiome and regulating immune response. Therefore, adjusting the diet can prevent and treat pouchitis. This article reviews the research progress of the influence of dietary factors on pouchitis.

OBJECTIVE:To establish the method for the simultaneous determination of contents of 4 heavy metal elements in cultured Cordyceps militaris. METHODS:The samples were digested by nitric acid heating method and then determined by ICP-MS including radio-frequency power of 1.15 kW,sampling depth of 65 nm,auxiliary gas flow rate of 1.0 L/min,cooling gas flow of 13.0 L/min,temperature of 25 ℃,automatic plasma. RESULTS:The linear ranges of arsenic(As),cadmium(Cd),mer-cury (Hg) and lead (Pb) were 0-20 μg/mL(r=0.9970),0-10 μg/mL(r=0.9995),0-5 μg/mL(r=0.9955),0-20 μg/mL(r=0.9960),respectively. Detectio limit were 0.128,0.003,0.002,0.004 mg. RSDs of precision,stability and reproducibility tests were all lower than 6.5%. Recoveries of them were 90.4%-100.6%(RSD=3.45%,n=9),94.3%-101.3%(RSD=2.93%,n=9), 90.0%-102.3%(RSD=4.03%,n=9),92.3%-103.0%(RSD=3.53%,n=9),respectively. CONCLUSIONS:The method is sim-ple,precise,stable,reproducible,and can be used for simultaneous determination of contents of 4 heavy metal elements in cul-tured C. militaris. The contents of As,Cd,Hg and Pb in 10 batches of sample are all in line with related national standards.

OBJECTIVE:To establish a method for simultaneous determination of 5 nucleoside from different parts of cultured Cordyceps militaris. METHODS:HPLC method was adopted. The determination was performed on InertSustain C18 column with mobile phase consisting of methanol-0.02 mol/L monobasic potassium phosphate solution (gradient elution) at the flow rate of 0.6 mL/min. The detection wavelength was set at 254 nm,the column temperature was 30 ℃. RESULTS:The linear ranges of uridine, inosine,guanosine,adenosine and cordycepin were 0.568-3.408 μg(r=0.9999),0.284-1.704 μg(r=0.9999),0.264-1.584 μg(r=0.9999),0.232-1.392 μg(r=0.9999)and 1.672-10.032 μg(r=0.9998),respectively. RSDs of precision,stability and repeatabili-ty tests were all lower than 3.0%. The recoveries were 98.2%-103.9%(RSD=1.97%,n=9),96.2%-101.6%(RSD=1.76%,n=9),96.7%-102.0%(RSD=1.94%,n=9),95.1%-99.4%(RSD=1.43%,n=9) and 95.6%-101.3%(RSD=1.82%,n=9). CON-CLUSIONS:The method is simple,precise,stable and repeatable,and can be used for simultaneous determination of 5 nucleoside from cultured C. militaris,the content of nucleosides are different in different parts of C. militaris.