Background/Aims: Early detection and effective prognosis prediction in patients with hepatocellular carcinoma (HCC) provide an avenue for survival improvement, yet more effective approaches are greatly needed. We sought to develop the detection and prognosis models with ultra-sensitivity and low cost based on fragmentomic features of circulating cell free mtDNA (ccf-mtDNA). Methods: Capture-based mtDNA sequencing was carried out in plasma cell-free DNA samples from 1168 participants, including 571 patients with HCC, 301 patients with chronic hepatitis B or liver cirrhosis (CHB/LC) and 296 healthy controls (HC). Results: The systematic analysis revealed significantly aberrant fragmentomic features of ccf-mtDNA in HCC group when compared with CHB/LC and HC groups. Moreover, we constructed a random forest algorithm-based HCC detection model by utilizing ccf-mtDNA fragmentomic features. Both internal and two external validation cohorts demonstrated the excellent capacity of our model in distinguishing early HCC patients from HC and highrisk population with CHB/LC, with AUC exceeding 0.983 and 0.981, sensitivity over 89.6% and 89.61%, and specificity over 98.20% and 95.00%, respectively, greatly surpassing the performance of alpha-fetoprotein (AFP) and mtDNA copy number. We also developed an HCC prognosis prediction model by LASSO-Cox regression to select 20 fragmentomic features, which exhibited exceptional ability in predicting 1-year, 2-year and 3-year survival (AUC=0.8333, 0.8145 and 0.7958 for validation cohort, respectively). Conclusions We have developed and validated a high-performing and low-cost approach in a large clinical cohort based on aberrant ccf-mtDNA fragmentomic features with promising clinical translational application for the early detection and prognosis prediction of HCC patients.

Background/Aims: Early detection and effective prognosis prediction in patients with hepatocellular carcinoma (HCC) provide an avenue for survival improvement, yet more effective approaches are greatly needed. We sought to develop the detection and prognosis models with ultra-sensitivity and low cost based on fragmentomic features of circulating cell free mtDNA (ccf-mtDNA). Methods: Capture-based mtDNA sequencing was carried out in plasma cell-free DNA samples from 1168 participants, including 571 patients with HCC, 301 patients with chronic hepatitis B or liver cirrhosis (CHB/LC) and 296 healthy controls (HC). Results: The systematic analysis revealed significantly aberrant fragmentomic features of ccf-mtDNA in HCC group when compared with CHB/LC and HC groups. Moreover, we constructed a random forest algorithm-based HCC detection model by utilizing ccf-mtDNA fragmentomic features. Both internal and two external validation cohorts demonstrated the excellent capacity of our model in distinguishing early HCC patients from HC and highrisk population with CHB/LC, with AUC exceeding 0.983 and 0.981, sensitivity over 89.6% and 89.61%, and specificity over 98.20% and 95.00%, respectively, greatly surpassing the performance of alpha-fetoprotein (AFP) and mtDNA copy number. We also developed an HCC prognosis prediction model by LASSO-Cox regression to select 20 fragmentomic features, which exhibited exceptional ability in predicting 1-year, 2-year and 3-year survival (AUC=0.8333, 0.8145 and 0.7958 for validation cohort, respectively). Conclusions We have developed and validated a high-performing and low-cost approach in a large clinical cohort based on aberrant ccf-mtDNA fragmentomic features with promising clinical translational application for the early detection and prognosis prediction of HCC patients.

Background/Aims: Early detection and effective prognosis prediction in patients with hepatocellular carcinoma (HCC) provide an avenue for survival improvement, yet more effective approaches are greatly needed. We sought to develop the detection and prognosis models with ultra-sensitivity and low cost based on fragmentomic features of circulating cell free mtDNA (ccf-mtDNA). Methods: Capture-based mtDNA sequencing was carried out in plasma cell-free DNA samples from 1168 participants, including 571 patients with HCC, 301 patients with chronic hepatitis B or liver cirrhosis (CHB/LC) and 296 healthy controls (HC). Results: The systematic analysis revealed significantly aberrant fragmentomic features of ccf-mtDNA in HCC group when compared with CHB/LC and HC groups. Moreover, we constructed a random forest algorithm-based HCC detection model by utilizing ccf-mtDNA fragmentomic features. Both internal and two external validation cohorts demonstrated the excellent capacity of our model in distinguishing early HCC patients from HC and highrisk population with CHB/LC, with AUC exceeding 0.983 and 0.981, sensitivity over 89.6% and 89.61%, and specificity over 98.20% and 95.00%, respectively, greatly surpassing the performance of alpha-fetoprotein (AFP) and mtDNA copy number. We also developed an HCC prognosis prediction model by LASSO-Cox regression to select 20 fragmentomic features, which exhibited exceptional ability in predicting 1-year, 2-year and 3-year survival (AUC=0.8333, 0.8145 and 0.7958 for validation cohort, respectively). Conclusions We have developed and validated a high-performing and low-cost approach in a large clinical cohort based on aberrant ccf-mtDNA fragmentomic features with promising clinical translational application for the early detection and prognosis prediction of HCC patients.

One of the basic questions in the aging field is whether there is a fundamental difference between the aging of lower invertebrates and mammals. A major difference between the lower invertebrates and mammals is the abundancy of noncoding RNAs, most of which are not conserved. We have previously identified a noncoding RNA Terc-53 that is derived from the RNA component of telomerase Terc. To study its physiological functions, we generated two transgenic mouse models overexpressing the RNA in wild-type and early-aging Terc-/- backgrounds. Terc-53 mice showed age-related cognition decline and shortened life span, even though no developmental defects or physiological abnormality at an early age was observed, indicating its involvement in normal aging of mammals. Subsequent mechanistic study identified hyaluronan-mediated motility receptor (Hmmr) as the main effector of Terc-53. Terc-53 mediates the degradation of Hmmr, leading to an increase of inflammation in the affected tissues, accelerating organismal aging. adeno-associated virus delivered supplementation of Hmmr in the hippocampus reversed the cognition decline in Terc-53 transgenic mice. Neither Terc-53 nor Hmmr has homologs in C. elegans. Neither do arthropods express hyaluronan. These findings demonstrate the complexity of aging in mammals and open new paths for exploring noncoding RNA and Hmmr as means of treating age-related physical debilities and improving healthspan.
Animals ; Mice ; RNA, Untranslated/metabolism* ; Aging/genetics* ; Mice, Transgenic ; Telomerase/metabolism* ; RNA/genetics* ; Hippocampus/metabolism* ; Humans ; Mice, Inbred C57BL

AIM: To investigate the effects of agkis-trodon halys venom anti-tumor component (AHVAC-) on the biological behavior of gastric cancer MKN-28 cells. METHODS: Gastric cancer MKN-28 cells were treated with the experimental concentrations (5, 10, 15 μg/mL) of AHAVC- for 24 h. Cell proliferation and toxicity assay (cell counting kit-8, CCK-8) was used to detect the inhibition rates of the cells in different concentrations of AHVAC-. The migration ability of the cells was evaluated by wound-healing and Transwell assay. The apoptosis were observed by laser confocal microscopy with annexin V-mCherry/DAPI double staining, and the apoptosis rates were analyzed by flow cytometry with annexin V-FITC/PI double fluorescence staining. The protein level of Caspease-3 was determined by Western blot. RESULTS: Compared with normal control group, the results of AHVAC- concentration groups showed that with the increase of AHVAC- concentration, the proliferative activity of MN-28 cells decreased gradually (P<0.01), the cell migration ability decreased gradually (P<0.01), and the cell apoptosis rate increased (P<0.05). The expression of apoptosis-related protein Caspease-3 was up-regulated (P<0.01). CONCLUSION: AHVAC- inhibits proliferation and migration of gastric cancer MSN-28 cells and induces apoptosis.

Objective:In order to develop a comprehensive hospital debt risk model,it is imperative to conduct an analysis of the present debt risk landscape within public hospitals,so as to furnish hospitals with recommendations to avert potential debt risks.Methods:A debt risk model was constructed based on 7 common factors through the extraction and standardization of data from 25 indicators,using 600 public hospitals in a specific province as samples,followed by factor analysis.Results:The model's calculation of the comprehensive score of public hospitals aligns with professional explanations and provides a more accurate representation of the factors influencing the debt risk level of public hospitals,such as financial investment level,hospital type,hierarchy,and others.Conclusion:In conclusion,the model is efficient.It is imperative to adopt a comprehensive perspective on the matter of hospital debt and efficiently address the associated financial risks faced by public hospitals by fostering collaboration among multiple stakeholders and upholding fundamental principles.

In order to explore the effects of Coptis detoxification powder on immune suppression and inflammatory response caused by heat stress in animals,Bama pigs were used as experimental animals to establish an animal model of heat stress,and the effects of Coptis detoxification powder on transcriptional activation of TLRs mRNA in PBMC induced by heat stress and its mediated ex-pression of inflammatory factors were studied in vivo.The animal model of heat stress was estab-lished by artificial climate greenhouse,Chinese medicine"Coptis detoxification powder"was pre-pared,and its inhibitory effect on heat stress and inflammation was studied by preventive and therapeutic administration.Real-time fluorescence quantitative PCR was used to determine the transcriptional changes of HSP70,TLRs and IL-17 mRNA,and ELISA was used to determine the dynamic changes of serum IL-2,IL-6 and TNF-α.Western blot and indirect immunofluorescence(IFA)were used to determine the nuclear translocation and protein expression of p65 phosphoryl-ated protein in NF-κβ inflammatory factor pathway.The transcription of PBMC TLRs mRNA and its mediated inflammatory factor expression and function in Bama pigs under heat stress were sys-tematically analyzed.The results showed that the experimental animal model of Bama pigs under heat stress was successfully established,and heat stress significantly up-regulated the mRNA tran-scription levels of HSP70,TLR2,TLR4 and inflammatory factor IL-17,resulting in significantly enhanced expressions of serum inflammatory factors IL-2,IL-6 and TNF-α.The NF-κβ pathway significantly promoted the level of p65 nuclear phosphorylation and the expression of cytoplasmic protein.The Chinese medicine"Coptis detoxification powder"could significantly reduce the upreg-ulation effect of heat stress conditions on the transcription levels of HSP70 and TLRs mRNA,thus inhibiting the mRNA transcription of inflammatory factor IL-17 and the expression of IL-2,IL-6 and TNF-α in serum,and significantly antagonizing the transcriptional activity of NF-κβ caused by heat stress.It showed good anti-heat stress effect.The anti-inflammatory effects of Chinese veteri-nary medicine prescription"Coptis detoxification powder",its pathway of action and its effectors were investigated,the expression and secretion levels of the regulatory factors related to the path-way associated with TLRs mRNA inhibition were analyzed,and the functional effects and mecha-nism of its clinical anti-heat stress and anti-inflammatory effects were clarified,which laid a data foundation for the promotion and application of this prescription and related mechanism research.

A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.
Animals ; Mice ; Peste-des-Petits-Ruminants/prevention & control* ; Antibodies, Monoclonal ; Reproducibility of Results ; Peste-des-petits-ruminants virus ; Antibodies, Viral ; Enzyme-Linked Immunosorbent Assay ; Goats

Irinotecan is an anticancer topoisomerase I inhibitor that acts as a prodrug of the active metabolite, SN-38. Unfortunately, the limited utility of irinotecan is attributed to its pH-dependent stability, short half-life and dose-limiting toxicity. To address this problem, a novel trivalent PEGylated prodrug (PEG-[Irinotecan]₃) has been synthesized and its full-profile pharmacokinetics, antitumor activity and toxicity compared with those of irinotecan. The results show that after intravenous administration to rats, PEG-[Irinotecan]₃ undergoes stepwise loss of irinotecan to form PEG-[Irinotecan]_3‒x (x = 1,2) and PEG-[linker] during which time the released irinotecan undergoes conversion to SN-38. As compared with conventional irinotecan, PEG-[Irinotecan]₃ displays extended release of irinotecan and efficient formation of SN-38 with significantly improved AUC and half-life. In a colorectal cancer-bearing model in nude mice, the tumor concentrations of irinotecan and SN-38 produced by PEG-[Irinotecan]₃ were respectively 86.2 and 2293 times higher at 48 h than produced by irinotecan. In summary, PEG-[Irinotecan]₃ displays superior pharmacokinetic characteristics and antitumor activity with lower toxicity than irinotecan. This supports the view that PEG-[Irinotecan]₃ is a superior anticancer drug to irinotecan and it has entered the phase II trial stage.

Objective:To investigate the relationship between serum 25(OH)D and SIRT4 levels and glycolipid metabolism in children with different levels of obesity.Methods:A total of 124 children with different levels of obesity who received treatment in Shaoxing Women's and Children's Health Care Hospital from February 2016 to February 2021 were included in this study. These children were divided into mild/moderate obesity group ( n = 76) and severe obesity group ( n = 48) according to body mass index. An additional 62 healthy children who concurrently received a physical examination were selected for controls. The general data of all children were collected. The relationship between the factors that affect obesity in children and serum 25(OH)D and SIRT4 levels and glycolipid metabolism was analyzed. Results:In the control, mild/moderate obesity, and severe obesity groups, body mass was (26.68 ± 4.98) kg, (33.24 ± 5.48) kg, (37.18 ± 5.88) kg, respectively; waist circumference was (56.12 ± 4.62) cm, (68.45 ± 5.20) cm, (79.34 ± 5.65) cm, respectively; hip circumference was (68.42 ± 5.08) cm, (72.45 ± 6.45) cm, (80.56 ± 6.95) cm, respectively; body mass index (BMI) was (15.90 ± 2.04) kg/m 2, (23.58 ± 2.45) kg/m 2, (25.89 ± 2.35) kg/m 2], respectively; fasting insulin (FINS) level was (26.65 ± 3.68) pmol/L, (34.82 ± 4.15) pmol/L, (48.56 ± 5.49) pmol/l, respectively; homeostasis model assessment of insulin resistance (HOMA-IR) was (1.06 ± 0.24), (2.12 ± 0.35), (3.84 ± 0.52), respectively; total cholesterol (TC) level was (2.21 ± 0.45) mmol/L, (4.14 ± 0.58) mmol/L, (5.96 ± 0.64) mmol/L, respectively; triacylglycerol (TG) level was (0.68 ± 0.16) mmol/L, (1.12 ± 0.24) mmol/L, (1.56 ± 0.35) mmol/L, respectively; low density lipoprotein cholesterol (LDL-C) was (2.68 ± 0.42) mmol/L, (2.10 ± 0.32) mmol/L, (1.41 ± 0.25) mmol/L, respectively; high density lipoprotein cholesterol (HDL-C) was (1.98 ± 0.42) mmol/L, (3.12 ± 0.51) mmol/L, (4.10 ± 0.56) mmol/L, respectively. There were significant differences in body mass, waist circumference, hip circumference, BMI, FINS, HOMA-IR, TC, TG, HDL-C, and LDL-C among the three groups ( F = 53.62, 280.42, 53.33, 303.44, 338.48, 755.71, 618.75, 165.81, 186.89, 251.42, all P < 0.001). Body mass, waist circumference, hip circumference, BMI, FINS, HOMA-IR, TC level, TG level, HDL-C level, and LDL-C level were lower in the control group than in the mild/moderate obesity group ( t = -7.28, -14.56, -4.00, -19.72, -6.49, -21.45, -12.36, 9.20, -14.12, all P < 0.05). Body mass, waist circumference, hip circumference, BMI, FINS, HOMA-IR, TC, TG, HDL-C and LDL-C were lower in the mild/moderate obesity group than in the severe obesity group ( t = -3.79, -10.98, -6.61, -5.19, -15.81, -22.02, -16.34, -8.30, 12.68, -10.03, all P < 0.05). Serum 25(OH)D [(60.52 ± 8.95) nmol/L vs. (49.88 ± 8.12) nmol /L, t = 7.31, P < 0.05] and SIRT4 [(1.98 ± 0.38) mmol/L vs. (1.06 ± 0.30) mmol/L, t = 15.89, P < 0.05] levels were significantly greater in the control group than in the mild/moderate obesity group. Serum 25(OH)D [(49.88 ± 8.12) nmol/L vs. (41.62 ± 7.50) nmol /L, t = 5.68, P < 0.05] and SIRT4 [(1.06 ± 0.30) mmol/L vs. (0.52 ± 0.15) mmol/L, t = 11.57, P < 0.05] levels were significantly greater in the mild/moderate obesity group than in the severe obesity group. Multiple linear regression analysis showed that body mass, waist circumference, hip circumference, FINS, HOMA-IR, TC, TG, and LDL were the positive influential factors of childhood obesity ( B = 0.170, 0.310, 0.403, 1.000, 3.464, 2.080, 2.656, 4.324); HDL, serum 25(OH)D and SIRT4 were the negative influential factors of childhood obesity ( B = -2.096, -0.156, -6.615). Pearson correlation analysis showed that serum 25(OH)D was significantly negatively correlated with FINS, HOMA-IR, TC, TG and LDL ( r = -0.20, -0.46, -0.30, -0.36, all P < 0.01), and significantly positively correlated with FPG and HDL ( r = 0.43, 0.77, both P < 0.01). Serum SIRT4 was negatively correlated with FINS, TC, TG, and LDL ( r = -0.48, -0.74, -0.61, -0.64, all P < 0.01), and positively correlated with FPG and HDL ( r = 0.21, 0.84, both P < 0.01). Conclusion:Serum 25(OH)D and SIRT4 levels decrease with the aggravation of obesity in children and are closely related to glycolipid metabolism. Therefore, early detection of obesity can reflect the degree of obesity and glycolipid metabolism in children.