BACKGROUND:Human periodontal ligament stem cells are potential functional cells for periodontal tissue engineering.However,long-term in vitro culture may lead to reduced stemness and replicative senescence of periodontal ligament stem cells,which may impair the therapeutic effect of human periodontal ligament stem cells. OBJECTIVE:To investigate the effect of oxymatrine on the stemness maintenance and osteogenic differentiation of periodontal ligament stem cells in vitro,and to explore the potential mechanism. METHODS:Periodontal ligament stem cells were isolated from human periodontal ligament tissues by tissue explant enzyme digestion and cultured.The surface markers of mesenchymal cells were identified by flow cytometry.Periodontal ligament stem cells were incubated with 0,2.5,5,and 10 μg/mL oxymatrine.The effect of oxymatrine on the proliferation activity of periodontal ligament stem cells was detected by CCK8 assay.The appropriate drug concentration for subsequent experiments was screened.Western blot assay was used to detect the expression of stem cell non-specific proteins SOX2 and OCT4 in periodontal ligament stem cells.qRT-PCR and western blot assay were used to detect the expression levels of related osteogenic genes and proteins in periodontal ligament stem cells. RESULTS AND CONCLUSION:(1)The results of CCK8 assay showed that 2.5 μg/mL oxymatrine significantly enhanced the proliferative activity of periodontal stem cells,and the subsequent experiment selected 2.5 μg/mL oxymatrine to intervene.(2)Compared with the blank control group,the protein expression level of SOX2,a stem marker of periodontal ligament stem cells in the oxymatrine group did not change significantly(P>0.05),and the expression of OCT4 was significantly up-regulated(P<0.05).(3)Compared with the osteogenic induction group,the osteogenic genes ALP,RUNX2 mRNA expression and their osteogenic associated protein ALP protein expression of periodontal ligament stem cells were significantly down-regulated in the oxymatrine+osteogenic induction group(P<0.05).(4)The oxymatrine up-regulated the expression of stemness markers of periodontal ligament stem cells and inhibited the bone differentiation of periodontal ligament stem cells,and the results of high-throughput sequencing showed that it may be associated with WNT2,WNT16,COMP,and BMP6.

AIM: To evaluate the application value of artificial intelligence-assisted systems in diagnosing diabetic retinopathy(DR)by Meta-analysis.METHODS: PubMed, Web of Science, Embase, Cochrane Library, CBM, CNKI, WanFang Data and VIP database were searched to collect relevant literature on the diagnostic value of artificial intelligence-assisted systems for DR from January 2019 to September 2024. The QUADAS-2 tool was used to evaluate the quality of the included studies, and Meta-analysis was performed using Stata 17.0 and Meta Disc 1.4 software.RESULTS: A total of 23 studies were included. The results of Meta-analysis showed that the pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio were 0.92(95% CI: 0.89-0.94), 0.94(95% CI: 0.91-0.96), 15.6(95% CI: 10.6-22.9), 0.09(95% CI: 0.07, 0.12), 174(95% CI: 112-271), respectively, and the area under the ROC curve(AUC)was 0.97(95% CI: 0.96-0.98). Meta-regression and subgroup analyses indicated that the heterogeneity of the studies originated from study type, patient type, patient source, and AI algorithm type. Deeks' funnel plot test suggested no significant publication bias(P=0.15), indicating that the results were robust.CONCLUSION: The artificial intelligence-assisted system demonstrates high diagnostic value for DR, and can be widely implemented in the early screening and diagnosis of DR.

Objective:To study the relationship between serum adiponectin level and early vascular aging (EVA).Methods:The cross-sectional study method was used. Six hundred and seventy-two subjects who completed health checkup from June to December 2023 in the Affiliated Hospital of Qingdao University were selected. The subjects were divided into the EVA group (237 cases) and the control group (435 cases) based on brachial-ankle pulse wave velocity (baPWV). According to the adiponectin tertiles method, the subjects were divided into low adiponectin subgroup (2.4 to 6.6 mg/L, 225 cases), medium adiponectin subgroup (6.7 to 9.1 mg/L, 227 cases) and high adiponectin subgroup (9.2 to 19.8 mg/L, 220 cases). The basic demographic information, past history and serological indexes were recorded. Univariate and multivariate binary Logistic regression analyses were used to analyze the risk factors for EVA, and multivariate Logistic regression was used to analyze the effect of adiponectin on EVA.Results:The male proportion, age, body mass index (BMI), systolic blood pressure, diastolic blood pressure, triglycerides (TG), fasting blood glucose (FBG), uric acid, glycated hemoglobin (HbA 1c), homocysteine, baPWV and alcohol history proportion in EVA group were significantly higher than those in control group: 64.98% (154/237) vs. 53.33% (232/435), 53 (47, 57) years old vs. 46 (39, 52) years old, (26.34 ± 3.37) kg/m 2 vs. (25.16 ± 3.91) kg/m 2, (132.27 ± 15.48) mmHg (1 mmHg = 0.133 kPa) vs. (117.30 ± 13.04) mmHg, (81.79 ± 11.04) mmHg vs. (71.93 ± 10.10) mmHg, 1.45 (1.03, 2.03) mmol/L vs. 1.08 (0.76, 1.65) mmol/L, 5.52 (5.03, 6.21) mmol/L vs. 5.14 (4.77, 5.56) mmol/L, (380.04 ± 96.43) μmol/L vs. (362.18 ± 94.94) μmol/L, 5.80 (5.50, 5.90)% vs. 5.70 (5.40, 5.82)%, 10.70 (9.01, 12.90) μmol/L vs. 9.96 (8.30, 12.20) μmol/L, 1 586 (1 511, 1 719) cm/s vs. 1 299 (1 215, 1 367) cm/s and 19.41% (46/237) vs. 13.56% (59/435), the adiponectin was significantly lower than that in control group: 7.00 (5.70, 8.75) mg/L vs. 8.40 (6.40, 10.60) mg/L, and there were statistical differences ( P<0.01 or <0.05). There were no statistical differences in total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), creatinine and smoking history proportion between two groups ( P>0.05). The male proportion, BMI, systolic blood pressure, diastolic blood pressure, TG, FBG, uric acid, creatinine, HbA 1c, homocysteine, EVA incidence, baPWV, smoking history proportion and alcohol history proportion in low adiponectin subgroup and medium adiponectin subgroup were significantly higher than those in high adiponectin subgroup, furthermore, the indexes except HbA 1c in low adiponectin subgroup were significantly higher than those in medium adiponectin subgroup, and there were statistical differences ( P<0.05); the HDL-C in low adiponectin subgroup and medium adiponectin subgroup was significantly lower than that in high adiponectin subgroup, furthermore, that in low adiponectin subgroup was significantly lower than that in medium adiponectin subgroup, and there were statistical differences ( P<0.05); there were no statistical differences in age, TC and LDL-C among the three subgroups ( P>0.05). Univariate binary Logistic regression analysis result showed that age, male, BMI, alcohol history, systolic blood pressure, diastolic blood pressure, TG, FBG, uric acid and HbA 1c were the risk factors for EVA ( P<0.01 or <0.05), while the adiponectin was a protective factor for EVA ( P<0.01). Multivariate binary Logistic regression analysis result showed that age, systolic blood pressure, TG and FBG were risk factors for EVA ( OR = 1.098, 1.066, 1.209 and 1.268; 95% CI 1.069 to 1.127, 1.050 to 1.082, 1.007 to 1.451 and 1.069 to 1.502; P<0.01 or <0.05), while adiponectin was a protective factor ( OR = 0.892, 95% CI 0.828 to 0.962, P<0.01). Multivariable Logistic regression analysis result showed that adiponectin consistently remained a protective factor for EVA across unadjusted, preliminary adjusted and fully adjusted covariate models ( OR = 0.553, 0.580 and 0.576; 95% CI 0.451 to 0.678, 0.440 to 0.764 and 0.435 to 0.763; P<0.01). Conclusions:The serum APN level is negatively correlated with the risk of EVA, which may be an independent protective factor for the EVA.

Objective:To study the relationship between serum adiponectin level and early vascular aging (EVA).Methods:The cross-sectional study method was used. Six hundred and seventy-two subjects who completed health checkup from June to December 2023 in the Affiliated Hospital of Qingdao University were selected. The subjects were divided into the EVA group (237 cases) and the control group (435 cases) based on brachial-ankle pulse wave velocity (baPWV). According to the adiponectin tertiles method, the subjects were divided into low adiponectin subgroup (2.4 to 6.6 mg/L, 225 cases), medium adiponectin subgroup (6.7 to 9.1 mg/L, 227 cases) and high adiponectin subgroup (9.2 to 19.8 mg/L, 220 cases). The basic demographic information, past history and serological indexes were recorded. Univariate and multivariate binary Logistic regression analyses were used to analyze the risk factors for EVA, and multivariate Logistic regression was used to analyze the effect of adiponectin on EVA.Results:The male proportion, age, body mass index (BMI), systolic blood pressure, diastolic blood pressure, triglycerides (TG), fasting blood glucose (FBG), uric acid, glycated hemoglobin (HbA 1c), homocysteine, baPWV and alcohol history proportion in EVA group were significantly higher than those in control group: 64.98% (154/237) vs. 53.33% (232/435), 53 (47, 57) years old vs. 46 (39, 52) years old, (26.34 ± 3.37) kg/m 2 vs. (25.16 ± 3.91) kg/m 2, (132.27 ± 15.48) mmHg (1 mmHg = 0.133 kPa) vs. (117.30 ± 13.04) mmHg, (81.79 ± 11.04) mmHg vs. (71.93 ± 10.10) mmHg, 1.45 (1.03, 2.03) mmol/L vs. 1.08 (0.76, 1.65) mmol/L, 5.52 (5.03, 6.21) mmol/L vs. 5.14 (4.77, 5.56) mmol/L, (380.04 ± 96.43) μmol/L vs. (362.18 ± 94.94) μmol/L, 5.80 (5.50, 5.90)% vs. 5.70 (5.40, 5.82)%, 10.70 (9.01, 12.90) μmol/L vs. 9.96 (8.30, 12.20) μmol/L, 1 586 (1 511, 1 719) cm/s vs. 1 299 (1 215, 1 367) cm/s and 19.41% (46/237) vs. 13.56% (59/435), the adiponectin was significantly lower than that in control group: 7.00 (5.70, 8.75) mg/L vs. 8.40 (6.40, 10.60) mg/L, and there were statistical differences ( P<0.01 or <0.05). There were no statistical differences in total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), creatinine and smoking history proportion between two groups ( P>0.05). The male proportion, BMI, systolic blood pressure, diastolic blood pressure, TG, FBG, uric acid, creatinine, HbA 1c, homocysteine, EVA incidence, baPWV, smoking history proportion and alcohol history proportion in low adiponectin subgroup and medium adiponectin subgroup were significantly higher than those in high adiponectin subgroup, furthermore, the indexes except HbA 1c in low adiponectin subgroup were significantly higher than those in medium adiponectin subgroup, and there were statistical differences ( P<0.05); the HDL-C in low adiponectin subgroup and medium adiponectin subgroup was significantly lower than that in high adiponectin subgroup, furthermore, that in low adiponectin subgroup was significantly lower than that in medium adiponectin subgroup, and there were statistical differences ( P<0.05); there were no statistical differences in age, TC and LDL-C among the three subgroups ( P>0.05). Univariate binary Logistic regression analysis result showed that age, male, BMI, alcohol history, systolic blood pressure, diastolic blood pressure, TG, FBG, uric acid and HbA 1c were the risk factors for EVA ( P<0.01 or <0.05), while the adiponectin was a protective factor for EVA ( P<0.01). Multivariate binary Logistic regression analysis result showed that age, systolic blood pressure, TG and FBG were risk factors for EVA ( OR = 1.098, 1.066, 1.209 and 1.268; 95% CI 1.069 to 1.127, 1.050 to 1.082, 1.007 to 1.451 and 1.069 to 1.502; P<0.01 or <0.05), while adiponectin was a protective factor ( OR = 0.892, 95% CI 0.828 to 0.962, P<0.01). Multivariable Logistic regression analysis result showed that adiponectin consistently remained a protective factor for EVA across unadjusted, preliminary adjusted and fully adjusted covariate models ( OR = 0.553, 0.580 and 0.576; 95% CI 0.451 to 0.678, 0.440 to 0.764 and 0.435 to 0.763; P<0.01). Conclusions:The serum APN level is negatively correlated with the risk of EVA, which may be an independent protective factor for the EVA.

Objective: To explore the effect of different concentrations of relaxin-2(RLN-2) on the proliferation and migration abilities of human immortalized keratinocytes(HaCaT cells). Methods: Methods HaCaT cells were cultured in media with different concentrations of RLN-2, and the cells were cultured in media without RLN-2 as the control group.The effect on cell proliferation was assessed by using the CCK-8 reagent, the cell migration ability was evaluated throughin vitrocell scratch assay, the cell cycle was examined by flow cytometry, and the expression levels of cell cycle proteins Cyclin B1 and Cyclin A2 were detected by Western blot. Results: After being cultured for 24 hours under RLN-2 concentration ranging from 10～100 ng/ml, HaCaT cells showed progressively increased proliferation and migration capabilities compared to the control group, with elevated expression levels of cell cycle proteins Cyclin B1 and Cyclin A2 and an increased proportion of cells in S and G2/M phases, peaking at 100 ng/ml. However, HaCaT cells cultured with 200 ng/ml of RLN-2 exhibited reduced proliferation and migration capabilities, decreased expression levels of Cyclin B1 and Cyclin A2, and a lower proportion of cells in S and G2/M phases compared to the 100 ng/ml group. Conclusion RLN-2 can enhance the migration ability of HaCaT cells within an appropriate concentration range and may also promote cell proliferation by increasing the expression of related cell cycle proteins and the proportion of cells in S and G2/M phases.

OBJECTIVE To screen potential adverse drug event （ADE） signals for the treatment of multiple sclerosis （MS） with ofatumumab， and to provide reference for the safe use of drugs in clinical practice. METHODS Using “ofatumumab” and the trade name “Kesimpta” as the search keywords， adverse event （AE） reports related to ofatumumab included in FDA Adverse Event Reporting System database from January 2009 to December 2023 were screened， and their reason contained the “multiple sclerosis”； ADE signal mining and analysis were conducted by reporting odds ratio method and proportional reporting ratio method. RESULTS A total of 21 759 eligible AE reports were selected， involving 62 449 AE cases； 27 system organ classes included general diseases and various reactions at the site of administration （15 021 cases）， neurological diseases （9 668 cases）， infectious and invasive diseases （5 967 cases）， injury， poisoning and surgical complications （4 952 cases）， musculoskeletal and connective tissue disorders （4 647 cases）. A total of 21 759 AE reports correspond to 606 ADE signals， including 234 ADE positive signals. A total of 107 ADE positive signals were not included in drug instruction of ofatumumab， including flu-like diseases， nasopharyngitis， cough， urinary tract infection， sore throat， insomnia， runny nose， anemia， hair loss， atrial fibrillation， and thrombocytopenia， etc. CONCLUSIONS In the process of using ofatumumab for MS， sufficient attention should be paid to ADE included in drug instructions. The ADE with strong signal strength screened in this study should also be paid special attention to， such as flu-like diseases， hemocytopenia， temperature intolerance， optic neuritis， and moyamoya disease. The increased risk of infection， cardiovascular disease， and potential damage to the respiratory and spiritual systems caused by ofatumumab can not be ignored.

In recent years, global outbreaks of infectious diseases, such as COVID-19, have triggered great concern about emerging infectious diseases. With the rapid development of next-generation sequencing technology and bioinformatic tools for viral metagenomics, there is now a widespread capability to detect and identify various known and unknown pathogenic microorganisms within both environmental and biological contexts. Furthermore, the continuous evolution of machine learning methods has led to the development and application of multiple rapid and highly accurate approaches for virus identification. Concurrently, owing to the continual progress in machine learning methods, several rapid and accurate virus identification techniques have been widely developed and applied. Therefore, this review aims to systematically summarize the key methodologies, frameworks, and the scope of applicability within the field of viral metagenomics, with a specific focus on virus identification and prediction. It could facilitate a deeper understanding of viral characteristics, identify potential novel pathogens, and provide technical support for the early prevention and control of infectious diseases.

Objective:To investigate the effects of botulinum toxin type A (BTX-A) and magnesium sulfate on the survival rate of random-pattern skin flaps (RSF) with different length-to-width ratios.Methods:Using a random number table method, 45 SD rats were divided into three groups: the saline group (Group A), the BTX-A group (Group B), and the magnesium sulfate group (Group C), with 15 rats in each group. Each group was further subdivided into three subgroups based on different length-to-width ratios of RSF (1∶1, 2∶1, 3∶1), with 5 rats in each subgroup. The preparation of the RSF involved using the midline of the rat’s back as the axis and the level 1 cm below the iliac crest line as the base, extending towards the head. The skin tissue was incised to the dorsal fascia layer, separating the subcutaneous tissue at the superficial layer of the deep fascia, while severing the blood vessels and their branches on both sides and at the base. After hemostasis, the flap was sutured in place. Immediately after surgery, 0.2 ml of saline, BTX-A (25 U/ml), or magnesium sulfate solution (50 mg/ml) was injected into the proximal, middle, and distal ends of the flap. On the seventh day post-surgery, the gross appearance of the flap was assessed, and the survival rate was calculated. The surviving flap tissue underwent hematoxylin and eosin (HE) staining to evaluate microvascular density (MVD) and the degree of vasodilation (including vessel outer diameter, inner diameter, and wall thickness). Immunohistochemistry was used to detect the expression level of vascular endothelial growth factor (VEGF). Statistical analysis was performed using GraphPad Prism 9.0.1 software, with data expressed as Mean ± SD. One-way ANOVA was used for multiple group comparisons, and Tukey’s test was used for pairwise comparisons.Results:On the seventh day post-surgery, flaps with a length-to-width ratio of 1∶1 healed well in all subgroups. In the case of flaps with a 2∶1 ratio, Group A exhibited partial necrosis at the distal end, characterized by blackened, non-elastic scabs and exudate. Groups B and C generally healed well. For flaps with a 3∶1 ratio, Group A exhibited extensive necrosis at both the middle and distal ends, with similar blackened, non-elastic scabs, non-bleeding cut sections, and exudate. Groups B and C showed only partial blackening at the distal end, with most areas healing effectively. The survival rates of flaps with a 1∶1 ratio did not show significant differences among the three groups ( P>0.05). Compared to Group A, Groups B and C had significantly higher survival rates for flaps with 2∶1 and 3∶1 ratios ( P<0.01), with no significant difference between Groups B and C ( P>0.05). HE staining indicated that as the length-to-width ratios increased, tissue edema and inflammatory cell infiltration also increased in all groups. Groups B and C had significantly reduced inflammatory changes compared to Group A, with a greater number of newly formed microvessels observed. Quantitative analysis revealed that MVD in Groups B and C was significantly higher than in Group A, regardless of the flap ratio ( P<0.05), with no significant difference between Groups B and C ( P>0.05). Vasodilation analysis showed that the outer diameter and wall thickness of vessels in Groups B and C were significantly greater than those in Group A ( P<0.05), with no significant difference between Groups B and C ( P>0.05). Immunohistochemical staining revealed that VEGF expression levels in Groups B and C were higher than in Group A, regardless of the flap ratio ( P<0.01). In flaps with a 1∶1 ratio, VEGF expression was higher in Group C than in Group B ( P<0.05), with no significant difference between the two groups for other flap ratios ( P>0.05). Conclusion:In RSF with length-to-width ratios of 2∶1 and 3∶1, subcutaneous injections of BTX-A or magnesium sulfate after replantation can promote the expansion and formation of blood vessels in the flap, increase the expression of VEGF, and improve the survival rate of the RSF.

Objective:To investigate the effects of botulinum toxin type A (BTX-A) and magnesium sulfate on the survival rate of random-pattern skin flaps (RSF) with different length-to-width ratios.Methods:Using a random number table method, 45 SD rats were divided into three groups: the saline group (Group A), the BTX-A group (Group B), and the magnesium sulfate group (Group C), with 15 rats in each group. Each group was further subdivided into three subgroups based on different length-to-width ratios of RSF (1∶1, 2∶1, 3∶1), with 5 rats in each subgroup. The preparation of the RSF involved using the midline of the rat’s back as the axis and the level 1 cm below the iliac crest line as the base, extending towards the head. The skin tissue was incised to the dorsal fascia layer, separating the subcutaneous tissue at the superficial layer of the deep fascia, while severing the blood vessels and their branches on both sides and at the base. After hemostasis, the flap was sutured in place. Immediately after surgery, 0.2 ml of saline, BTX-A (25 U/ml), or magnesium sulfate solution (50 mg/ml) was injected into the proximal, middle, and distal ends of the flap. On the seventh day post-surgery, the gross appearance of the flap was assessed, and the survival rate was calculated. The surviving flap tissue underwent hematoxylin and eosin (HE) staining to evaluate microvascular density (MVD) and the degree of vasodilation (including vessel outer diameter, inner diameter, and wall thickness). Immunohistochemistry was used to detect the expression level of vascular endothelial growth factor (VEGF). Statistical analysis was performed using GraphPad Prism 9.0.1 software, with data expressed as Mean ± SD. One-way ANOVA was used for multiple group comparisons, and Tukey’s test was used for pairwise comparisons.Results:On the seventh day post-surgery, flaps with a length-to-width ratio of 1∶1 healed well in all subgroups. In the case of flaps with a 2∶1 ratio, Group A exhibited partial necrosis at the distal end, characterized by blackened, non-elastic scabs and exudate. Groups B and C generally healed well. For flaps with a 3∶1 ratio, Group A exhibited extensive necrosis at both the middle and distal ends, with similar blackened, non-elastic scabs, non-bleeding cut sections, and exudate. Groups B and C showed only partial blackening at the distal end, with most areas healing effectively. The survival rates of flaps with a 1∶1 ratio did not show significant differences among the three groups ( P>0.05). Compared to Group A, Groups B and C had significantly higher survival rates for flaps with 2∶1 and 3∶1 ratios ( P<0.01), with no significant difference between Groups B and C ( P>0.05). HE staining indicated that as the length-to-width ratios increased, tissue edema and inflammatory cell infiltration also increased in all groups. Groups B and C had significantly reduced inflammatory changes compared to Group A, with a greater number of newly formed microvessels observed. Quantitative analysis revealed that MVD in Groups B and C was significantly higher than in Group A, regardless of the flap ratio ( P<0.05), with no significant difference between Groups B and C ( P>0.05). Vasodilation analysis showed that the outer diameter and wall thickness of vessels in Groups B and C were significantly greater than those in Group A ( P<0.05), with no significant difference between Groups B and C ( P>0.05). Immunohistochemical staining revealed that VEGF expression levels in Groups B and C were higher than in Group A, regardless of the flap ratio ( P<0.01). In flaps with a 1∶1 ratio, VEGF expression was higher in Group C than in Group B ( P<0.05), with no significant difference between the two groups for other flap ratios ( P>0.05). Conclusion:In RSF with length-to-width ratios of 2∶1 and 3∶1, subcutaneous injections of BTX-A or magnesium sulfate after replantation can promote the expansion and formation of blood vessels in the flap, increase the expression of VEGF, and improve the survival rate of the RSF.

In recent years, global outbreaks of infectious diseases, such as COVID-19, have triggered great concern about emerging infectious diseases. With the rapid development of next-generation sequencing technology and bioinformatic tools for viral metagenomics, there is now a widespread capability to detect and identify various known and unknown pathogenic microorganisms within both environmental and biological contexts. Furthermore, the continuous evolution of machine learning methods has led to the development and application of multiple rapid and highly accurate approaches for virus identification. Concurrently, owing to the continual progress in machine learning methods, several rapid and accurate virus identification techniques have been widely developed and applied. Therefore, this review aims to systematically summarize the key methodologies, frameworks, and the scope of applicability within the field of viral metagenomics, with a specific focus on virus identification and prediction. It could facilitate a deeper understanding of viral characteristics, identify potential novel pathogens, and provide technical support for the early prevention and control of infectious diseases.