BACKGROUND:Bone tissue loss caused by tumors and trauma can have an adverse effect on postoperative rehabilitation.Therefore,scaffold materials are usually implanted during treatment.However,the existing implant materials are relatively simple and lack antibacterial properties.Early implantation may lead to iatrogenic autoinfection and have an adverse effect on osteogenesis.OBJECTIVE:To construct a KR-12-a5 polypeptide-nanohydroxyapatite-small intestinal submucosa composite scaffold and evaluate its feasibility as a material for promoting bone defect repair.METHODS:The small intestinal submucosa scaffold and the small intestinal submucosa scaffold containing 25,50,and 100 mg/mL nanohydroxyapatite(referred to as nHA-SIS scaffold)were prepared by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysuccinimide cross-linking method.The appropriate scaffold was screened for subsequent experiments by mechanical property testing.The antibacterial properties of KR-12-a5 polypeptide solution against Staphylococcus aureus,Streptococcus gordonii,and Fusobacterium nucleatum were detected.The nHA-SIS scaffolds were immersed in 250,500,and 1 000 μg/mL KR-12-a5 peptide solutions for 24 hours,and then freeze-dried to obtain peptide-loaded nanohydroxyapatite-porcine small intestinal submucosa composite scaffolds(denoted as P-nHA-SIS scaffolds).The sustained-release properties of the three groups of scaffolds were characterized.The nHA-SIS scaffolds and the three groups of P-nHA-SIS scaffolds were co-cultured with Staphylococcus aureus,Streptococcus gordonii,and Fusobacterium nucleatum for 24 hours or 48 hours.The scaffolds with strong antibacterial ability were screened by live and dead bacteria staining and scanning electron microscopy for subsequent experiments.The degradation properties and water absorption rates of the uncross-linked small intestinal submucosa scaffolds,cross-linked small intestinal submucosa scaffolds,nHA-SIS scaffolds,and P-nHA-SIS scaffolds were characterized.The extracts of cross-linked small intestinal submucosal scaffolds,nHA-SIS scaffolds,and P-nHA-SIS scaffolds were co-cultured with MC3T3-E1 cells.CCK-8 assay and live-dead cell staining were performed.The effects of the extracts of the three scaffolds on the migration of MC3T3-E1 cells were detected by Transwell chamber assay.RESULTS AND CONCLUSION:(1)The elastic modulus and compressive strength of 25,50,and 100 mg/mL nHA-SIS scaffolds were higher than those of small intestinal submucosal scaffolds(P＜0.05),among which the elastic modulus and compressive strength of 25 mg/mL nHA-SIS scaffolds were the highest,and this group of scaffolds were selected for subsequent experiments to load peptides.(2)KR-12-a5 peptide had strong antibacterial activity against common bacteria in bone defects(Staphylococcus aureus,Streptococcus gordonii,and Fusobacterium nucleatum).The three groups of P-nHA-SIS scaffolds all had sustained release properties.With the increase of peptide mass concentration,the antibacterial property of P-nHA-SIS scaffold was enhanced.Among them,the P-nHA-SIS scaffold loaded with 500 μg/mL peptide had achieved a satisfactory antibacterial effect,and this group of scaffolds would be selected in the future.(3)The degradation rate of the three groups of cross-linked scaffolds was lower than that of the uncross-linked scaffolds,and the water absorption rate was greater than that of the uncross-linked scaffolds.P-nHA-SIS scaffolds could promote the proliferation and migration of MC3T3-E1 cells without affecting the activity of MC3T3-E1 cells.(4)The results show that P-nHA-SIS scaffolds have strong antibacterial properties and the ability to promote the proliferation and migration of MC3T3-E1 cells,and are expected to be used in bone defect repair.

BACKGROUND:Bone tissue loss caused by tumors and trauma can have an adverse effect on postoperative rehabilitation.Therefore,scaffold materials are usually implanted during treatment.However,the existing implant materials are relatively simple and lack antibacterial properties.Early implantation may lead to iatrogenic autoinfection and have an adverse effect on osteogenesis.OBJECTIVE:To construct a KR-12-a5 polypeptide-nanohydroxyapatite-small intestinal submucosa composite scaffold and evaluate its feasibility as a material for promoting bone defect repair.METHODS:The small intestinal submucosa scaffold and the small intestinal submucosa scaffold containing 25,50,and 100 mg/mL nanohydroxyapatite(referred to as nHA-SIS scaffold)were prepared by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysuccinimide cross-linking method.The appropriate scaffold was screened for subsequent experiments by mechanical property testing.The antibacterial properties of KR-12-a5 polypeptide solution against Staphylococcus aureus,Streptococcus gordonii,and Fusobacterium nucleatum were detected.The nHA-SIS scaffolds were immersed in 250,500,and 1 000 μg/mL KR-12-a5 peptide solutions for 24 hours,and then freeze-dried to obtain peptide-loaded nanohydroxyapatite-porcine small intestinal submucosa composite scaffolds(denoted as P-nHA-SIS scaffolds).The sustained-release properties of the three groups of scaffolds were characterized.The nHA-SIS scaffolds and the three groups of P-nHA-SIS scaffolds were co-cultured with Staphylococcus aureus,Streptococcus gordonii,and Fusobacterium nucleatum for 24 hours or 48 hours.The scaffolds with strong antibacterial ability were screened by live and dead bacteria staining and scanning electron microscopy for subsequent experiments.The degradation properties and water absorption rates of the uncross-linked small intestinal submucosa scaffolds,cross-linked small intestinal submucosa scaffolds,nHA-SIS scaffolds,and P-nHA-SIS scaffolds were characterized.The extracts of cross-linked small intestinal submucosal scaffolds,nHA-SIS scaffolds,and P-nHA-SIS scaffolds were co-cultured with MC3T3-E1 cells.CCK-8 assay and live-dead cell staining were performed.The effects of the extracts of the three scaffolds on the migration of MC3T3-E1 cells were detected by Transwell chamber assay.RESULTS AND CONCLUSION:(1)The elastic modulus and compressive strength of 25,50,and 100 mg/mL nHA-SIS scaffolds were higher than those of small intestinal submucosal scaffolds(P＜0.05),among which the elastic modulus and compressive strength of 25 mg/mL nHA-SIS scaffolds were the highest,and this group of scaffolds were selected for subsequent experiments to load peptides.(2)KR-12-a5 peptide had strong antibacterial activity against common bacteria in bone defects(Staphylococcus aureus,Streptococcus gordonii,and Fusobacterium nucleatum).The three groups of P-nHA-SIS scaffolds all had sustained release properties.With the increase of peptide mass concentration,the antibacterial property of P-nHA-SIS scaffold was enhanced.Among them,the P-nHA-SIS scaffold loaded with 500 μg/mL peptide had achieved a satisfactory antibacterial effect,and this group of scaffolds would be selected in the future.(3)The degradation rate of the three groups of cross-linked scaffolds was lower than that of the uncross-linked scaffolds,and the water absorption rate was greater than that of the uncross-linked scaffolds.P-nHA-SIS scaffolds could promote the proliferation and migration of MC3T3-E1 cells without affecting the activity of MC3T3-E1 cells.(4)The results show that P-nHA-SIS scaffolds have strong antibacterial properties and the ability to promote the proliferation and migration of MC3T3-E1 cells,and are expected to be used in bone defect repair.

To evaluate the therapeutic effect of baicalein topical preparation on atopic dermatitis, we first constructed two atopic dermatitis-like mouse models induced by calcipotriol and 1-fluoro-2,4-dinitrobenzene to assess their therapeutic effect with skin tissue staining and other experiments. It was found that topical preparation of baicalein could alleviate epidermal thickening of diseased skin tissues, repair damaged skin barrier proteins, and inhibit T helper 2 cells (Th2) infiltration and mast cell infiltration and activation in lesional sites. Cyberpharmacology was utilized to analyze whether baicalein could treat atopic dermatitis by interfering with multiple pathogenesis-associated pathways. Results indicated that baicalein reduced the mRNA levels of inflammatory factors and inhibited the phosphorylation of NF-κB p65 and STAT1 proteins in keratinocyte cells. Together, the topical preparation of baicalein may be effective in alleviating atopic dermatitis-like symptoms in mice by down-regulating the phosphorylation level of NF-κB in keratinocytes, thereby decreasing the expression of inflammatory factors in keratinocytes, which provides an idea and a theoretical basis for the topical preparation of baicalein for the treatment of inflammatory skin diseases such as atopic dermatitis.

To investigate the effects of Wogonin (WO) on learning and memory impairment, Aβ1-42 was injected intracerebroventricularly to induced a mouse learning and memory impairment model, and D-galactose was injected intraperitoneally to induced a mouse acute aging model. Mice were administered WO (75, 150, or 300 mg/kg) by oral gavage for 28 consecutive days. Cognitive function was assessed using the Morris water maze (MWM), novel object recognition (NOR), and open field tests (OFT). In the Aβ1-42 model, WO treatment (150 and 300 mg/kg) significantly improved the recognition index in the NOR test, while the 150 mg/kg group showed increased target quadrant preference in the MWM test. No changes in the total distance traveled in OFT. In the D-galactose aging model, the 150 mg/kg WO group exhibited increased platform crossings in the MWM test, and all WO doses (75, 150, and 300 mg/kg) enhanced target quadrant preference, with no alterations in spontaneous movement. Western blot analysis revealed that WO significantly attenuated hippocampal apoptosis in both models. These findings suggest that WO ameliorates learning and memory impairment associated with Alzheimer’s disease and aging.

Objective: To analyze the characteristics of alcohol poisoning cases among minors receiving pre hospital 120 emergency services in Zhengzhou, providing evidence for regional management strategies of alcohol poisoning among minors. Methods: A retrospective study was conducted on 1 630 alcohol poisoning cases (aged 0-18 years) from Zhengzhou s 120 emergency system during 2017-2019 and 2023. Data on gender, age, occurrence timeframes were analyzed using t-test and χ 2 test for intergroup comparisons. Results: Annual cases were 291 (2017), 353 (2018), 483 (2019), and 503 (2023). Compared with 2017, male alcohol poisoning cases increased by 66.94% while female cases surged 104.35% by 2023. The peak incidence of alcohol poisoning among minors occurred among 16-18 year olds (85.40%), followed by 13-15 year olds (13.74%). Most cases clustered between 21:01-03:00 (60.43%), with male cases peaking at 22:01-23:00 (12.73%) and female cases peaking at 02:01-03:00 ( 11.25 %). Between 00:01-03:00, male cases progressively decreased while female cases increased. Severity distribution showed 355 mild cases (21.78%), 1 035 moderate cases (63.50%), and 240 severe cases (14.72%). Conclusions Zhengzhou region has experienced sustained growth in underage alcohol poisoning cases, predominantly occurring from evening to early morning with moderate severity, female cases demonstrate faster growth rates. Multifaceted regulatory measures should be implemented to strengthen supervision of underage drinking behaviors.

Objective To establish an online gel permeation chromatography-gas chromatography/mass spectrometry(GPC-GCMS)method for the determination of etomidate in blood.Methods Blood samples were extracted with acetonitrile,dehydrated with anhydrous sodium sulfate,purified with an online GPC system,and then analyzed by GCMS using large volume injection.The external standard curve method was used for quantification.Results The detection limit of etomidate was 0.002 μg/mL,with good linearity in the concentration range of 0.01～5 μg/mL and a correlation coefficient of 0.9992.The extraction recovery rates ranged between 90%and 120%.In an actual case,the content of etomidate detected in the blood sample was 0.024 μg/mL.Conclusion This method is fast,simple,accurate,and stable,providing a reliable basis for the determination of etomidate content in blood.

This study investigated the effects and underlying mechanisms of the luteolin-naringenin combination(LN)on liver injury induced by acetaminophen(APAP).Forty-eight Kunming mice were randomly allocated into six groups:a normal control group,an APAP-induced liver injury model group,a positive drug treatment group,and three LN treatment groups with low,medium,and high doses.After the final drug administration,the mice were fasted for 12 hours prior to eu-thanasia for sample collection.Serum transaminase activity,oxidative stress indices,and hematoxy-lin-eosin(HE)staining were assessed to evaluate the effects of LN on APAP-induced hepatic inju-ry.Additionally,Western blot analysis was conducted to examine the expression levels of sphingo-sine kinase 1(SPHK1)and endoplasmic reticulum stress(ERS)-related proteins,thereby elucida-ting the potential mechanisms by which LN mitigates APAP-induced liver injury.The results dem-onstrated that varying concentrations of LN effectively ameliorated serum aminotransferase activi-ty and oxidative stress levels induced by APAP in a dose-dependent manner.Histopathological ex-amination via HE staining revealed significant improvement in APAP-induced liver tissue injury following treatment with different concentrations of LN.Furthermore,Western blot analysis indi-cated that the protein expressions of SPHK1,CHOP,p-IRE1α,ATF6,p-PERK,p-eIF2α,and ATF4 were markedly reduced after administration of various concentrations of LN.The results demonstrate that LN exhibits a significant protective effect against APAP-induced liver injury by inhibiting the SPHK1-mediated aberrant expression of ERS-related molecules.This study high-lights the importance of targeting SPHK1 in the treatment of APAP liver injury and provides a no-vel therapeutic approach through the multi-target and multi-pathway combination of monomers.

BACKGROUND:THZ1,an inhibitor of cyclin-dependent kinase 7,has been shown to inhibit the proliferation of a variety of tumor cells,but whether THZ1 can affect the stemness of glioma stem cells through the Wnt/β-catenin signaling pathway remains unclear.OBJECTIVE:To investigate the effect of THZ1 on stemness of glioma cell U87 and its mechanism.METHODS:U87 adherent cells were cultured to form stem cell mammospheres.The expressions of stemness related proteins were verified by western blot assay.The effect of THZ1 on half maximal inhibitory concentration(IC50)of U87 cells was determined by Cell Counting Kit-8(CCK-8)assays.The effects of THZ1 on proliferation and migration of U87 cells were determined by cell colony-formation assays,cell wound healing assays,and Transwell migration assays.The effect of THZ1 treatment on mammosphere forming rate and mammosphere size of U87 stem cells was analyzed.Stemness associated proteins CD133,ABCG2,Nanog,OCT4,SOX2,epithelial-mesenchymal transformation-related proteins E-cadherin,N-cadherin,Occludin,Snail,and Wnt/β-catenin pathway associated proteins Axin1,β-Catenin,WNT-5A,GSK3β,Cyclind-1,and C-myc were measured by western blot assay.RESULTS AND CONCLUSION:(1)Compared with adherent cells,the expressions of stemness related proteins Nestin,CD133,ABCG2,Nanog,OCT4,and SOX2 were significantly increased.(2)Compared with the control group,THZ1 decreased the proliferation and migration of U87 cells.(3)THZ1 inhibited the mammosphere forming rate and mammosphere size of U87 stem cells.(4)After THZ1 treatment,the expression of N-cadherin and Snail decreased,while the protein expression of E-cadherin and Occludin increased.(5)THZ1 treatment decreased the expression of Wnt/β-catenin pathway related proteins Axin1,β-Catenin,Wnt-5A,GSK3β,Cyclind-1,and C-myc in U87 stem cells.It is concluded that THZ1 can suppress the proliferation and migration of U87 cells,and inhibit the mammosphere forming ability,stemness related protein expression,and epithelial-mesenchymal transformation ability of U87 stem cells by down-regulating the expression of Wnt/β-catenin signaling pathway related molecules.

Objective To observe the cardioprotection of Rhizoma Polygonati(RP)in natural aging rat and its molecular mechanism.Methods 16 SD rats aged 2 months were selected as the young group,another 64 SD rats aged 18 months were randomly divided into 4 groups:an old group,a RP-low,a RP-medium and a RP-high dose group,with 16 rats in each group.The RP-low,RP-medium and RP-high dose groups were administered of RP(1,2,4 g·kg-1)by gavage,the old and young groups were given the same amount of distilled water once a day for 12 weeks.At week 4 and 12,eight rats from each group were sacrificed under anesthesia and hearts were isolated.Then cardiac weight index(CI)were measured and the histopathologic observation was assessed.The contents of total antioxidant capacity(T-AOC),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD)and malondialdehyde(MDA)were measured,and the expression of ataxia-telangiectasia mutated gene(ATM),ATM and Rad3-related kinase(ATR)-related protein in cardiac tissue were detected.Results With the increase of weeks of age,the fibrosis area in the old group was increased.The content of GSH-Px was decreased while MDA was increased;the expressions of Chk1 and Chk2 were also up-regulated(P<0.05).At week 4 or 12,compared to the young group,the CI was reduced,and the contents of T-AOC,SOD,GSH-Px were decreased while MDA was increased;the expression of ATM,ATR-related proteins were all significantly up-regulated in cardiac tissue of old group(P<0.05).After RP treatment,the CI was elevated,the structure of cardiomyocytes were intact and the myocardial fibers were arranged neatly,the fibrosis area was lowered.The contents of T-AOC,GSH-Px,SOD were increased while MDA was decreased.The expression of ATM,ATR-related proteins were significantly down-regulated in cardiac tissue(P<0.05).Conclusion RP could protect the heart in natural aging rats,and its mechanism may be related to the regulation of DNA damage response ATM,ATR signaling pathway.

Objective To compare the efficacy of artificial intelligence(AI)diagnostic group and artificial reading group in the diagnosis for osteoporotic vertebral compression fractures.Methods From January 2023 to December 2023,80 patients with osteoporotic vertebral compression fractures and 20 patients without fractures but with nonspecific low back pain were included in the study.According to the patient's computed tomography(CT)image,the AI software diagnosis and physicians of different seniority(one senior physician,one intermediate physician and one junior physician)diagnosis were performed.The diagnostic efficacy of different detection methods was compared.Results The sensitivity,specificity,positive predictive value,negative predictive value and area under the receiver operating characteristic(ROC)curve(AUC)and Kappa value of each group were as follows:AI image interpretation:0.975,0.900,0.975,0.900,0.938,0.875;senior physician:0.950,0.900,0.974,0.818,0.925,0.819;intermediate physician:0.825,0.850,0.957,0.548,0.837,0.560;and junior physician:0.750,0.750,0.923,0.429,0.751,0.390.Conclusion The diagnostic performance of AI was comparable to that of senior physician,and significantly higher than that of intermediate and primary physicians.