BACKGROUND:Tissue engineering is considered an ideal treatment for growth plate regeneration.However,most of the current research on regenerative tissue engineering is the traditional scaffold-based strategy.As the limitations of traditional scaffolds are gradually revealed,the research direction is gradually diversifying. OBJECTIVE:To summarize the application of scaffold-based and scaffold-free strategies in the treatment of growth plate cartilage regeneration and their respective advantages and disadvantages. METHODS:The relevant articles were searched from PubMed,Wiley,and Elsevier.The search terms were"growth plate injury,regeneration,tissue engineering,scaffold,scaffold-free,biomimetic,cartilage"in English.The time was limited from 1990 to 2023.Finally,104 articles were included for review. RESULTS AND CONCLUSION:The biomimetic strategy is to reduce the cell composition,biological signals and unique mechanical properties of each region to the greatest extent by simulating the unique organizational structure of the growth plate,so as to build a biomimetic microenvironment that can promote tissue regeneration.Therefore,the design of a biomimetic scaffold is to simulate the original growth plate as far as possible in terms of composition,structure and mechanical properties.Although some results have been achieved,there is still the problem of the unstable regeneration effect.The scaffold-free strategy believes that the limitations of scaffolds will have adverse effects on regenerative therapy.Therefore,the design of scaffold-free constructs relies as much as possible on the ability of cells to generate and maintain extracellular matrix without interfering with cell-cell signals or introducing exogenous substances.However,there are some problems,such as poor stability,low mechanical strength and greater difficulty in operation.Biomimetic strategy and scaffold-free strategy have different emphases,advantages and disadvantages,but they both have positive effects on growth plate cartilage regeneration.Therefore,subsequent studies,whether adopting a biomimetic strategy or a scaffold-free strategy,will focus on the continuous optimization of existing technologies in order to achieve effective growth plate cartilage regeneration therapy.

BACKGROUND:Nowadays,posterior cruciate ligament injury caused by a sports injury or vehicle injury is more common than people think.Posterior cruciate ligament reconstruction is one of the main treatment methods,but there are still a lot of controversies about the surgical method and ligament selection of posterior cruciate ligament reconstruction. OBJECTIVE:To comprehensively analyze the global application trend of posterior cruciate ligament reconstruction and identify promising research hotspots of posterior cruciate ligament reconstruction based on bibliometrics and visual analysis. METHODS:Publications(articles and reviews)related to posterior cruciate ligament reconstruction from 2000 to 2022 were retrieved from the Web of Science(WOS).The country,institution,publication year,author,journal,average citations per item,H index,title,keywords of publication,and the top 25 cited articles were extracted and analyzed in detail.The VOSviewer/citespace/Pajek software was used to analyze the co-occurrence result of keywords to predict the hotspots of posterior cruciate ligament reconstruction. RESULTS AND CONCLUSION:A total of 664 articles were included.(1)In the past 22 years,the number of posterior cruciate ligament reconstruction articles has shown an increasing trend in general.The top 3 countries(the USA,China,and South Korea)accounted for 65.51%of all articles published.The USA has the largest number of publications.The University of Pittsburgh is the largest contributor.Knee Surgery Sports Traumatol Arthrosc and American Journal of Sports Medicine are the most influential journals.Laprade,Robert F.is the professor who has published the most articles in the field of posterior cruciate ligament reconstruction,and Fanelli,GC is the professor who has the highest total chain strength in the field of posterior cruciate ligament reconstruction.(2)The research direction can be divided into the following five clusters:"posterior cruciate ligament anatomical and biomechanical studies","posterior cruciate ligament reconstruction prognosis,outcome,and complications","posterior cruciate ligament reconstruction surgical method and tendon selection","surgical technique",and"posterior cruciate ligament tear combined with multiple ligament injury".(3)It is concluded that in terms of the trend of previous years,an increasing number of articles related to posterior cruciate ligament reconstruction will be published in the future.The USA is a world leader in the field of posterior cruciate ligament reconstruction.China and South Korea presented great potential in this area.Anatomical and biomechanical research of posterior cruciate ligament and posterior cruciate ligament reconstruction methods and the selection of tendons may be the future hotspots in the field of posterior cruciate ligament reconstruction.

Objective: To explore the relationship between dietary behavior, dietary frequency and central precocious puberty (CPP) among children in Wuhan, so as to provide a scientific basis for elucidating the possible and related dietary factors leading to precocious puberty. Methods: From February to December 2023, 100 children with CPP and 100 healthy children were collected in Wuhan Children s Hospital for a case control study. Questionnaires and interviews were used to collect demographic information, child lifestyle, as well as dietary behavior and dietary frequency in the past six months. Conditional Logistic regression model was used to analyze the relationship of children s dietary behavior and dietary frequency with CPP. Results: No associations were found between three dietary behavior, including emotional eating, restrictive eating, and external eating, with CPP at baseline ( P >0.05). In the association analysis of dietary frequency and CPP, the risk of CPP was higher in those who consumed red meat >5 times/week ( OR =1.93, 95% CI =1.01-3.68), carbonated beverages ≥4 times/month ( OR =2.70, 95% CI =1.03-7.08), fruit juices ≥4 times/month ( OR =2.31, 95% CI =1.02-5.25), and nutritional supplementation ( OR =2.77, 95% CI =1.47- 5.22 ), whereas the risk of CPP was lower in those who consumed nuts ≥4 times/month ( OR =0.21, 95% CI =0.08-0.57) ( P < 0.05 ). Conclusion Consumption of higher amounts of red meat, high frequency of carbonated and juice based sugary drinks, and nutrient supplementation may be contributing factors to the development of CPP, whereas regular consumption of nuts is less likely to be associated with the development of CPP.

Objective We aimed to study the protective effect of total glucosides of paeony (TGP) on chemical liver injury with pattern of liver yin deficiency in rats and determine whether it exerts these effects through the phosphoinositide 3-kinase(PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway.Methods Forty male Sprague-Dawley rats were randomly divided into the following four groups: (i) the blank group, (ii) the model group, (iii) the Yiguan Jian group, and (iv) the TGP group (10 rats per group). For 6 weeks, rats in all groups except for the blank group were injected intraperitoneally with 20％ carbon tetrachloride olive oil solution and were given thyroid tablets (30mg/kg) by gavage in order to establish the model of chemical liver injury with pattern of liver yin deficiency. During the modeling period, rats in the blank and model groups were gavaged daily with distilled water, while rats in the Yiguan Jian group and the TGP group were gavaged with 635mg/kg of Yiguan Jian decoction and 50mg/kg of TGP suspension, respectively. During the administration period, the body weight and rectal temperature of rats were measured every 2 weeks. After the administration, 24-hour food intake, 24-hour water intake, and moisture capacity in tongue surface were recorded. Serum cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and interleukin-1 receptor antagonist (IL-1Ra) contents were measured by ELISA and the cAMP/cGMP ratio was calculated. Changes in the serum levels of alanine aminotransferase (ALT), alanine transaminase (AST), gamma-glutamyl transpeptidase (γ-GT), alkaline phosphatase (ALP), total bilirubin (TBIL), and total bile acids (TBA) were detected using colorimetric method. Masson staining was performed to observe the degree of liver fibrosis and to calculate the relative collagen area. Real-time PCR and Western blotting were conducted to determine the PI3K, AKT, and mTOR mRNA and protein expression levels in rat liver.Results Compared with the blank group, rats in the model group had a lower body weight at weeks 2, 4, and 6, a higher rectal temperature at weeks 2, 4, and 6, and a higher 24-hour food intake; the cAMP level was elevated, the cGMP level was decreased, and the cAMP/cGMP ratio was elevated; the contents of IL-1, IL-6, and TNF-α were elevated, and the contents of IL-10 and IL-1Ra were decreased; ALT, AST, γ-GT, ALP, TBIL, and TBA levels were elevated; the percentage of collagen area in the liver was increased; and the mRNA and protein phosphorylation levels of PI3K, AKT, and mTOR were elevated ( P<0.05). Compared with the model group, rats in the TGP group showed a decrease in rectal temperature at weeks 2, 4, and 6, a decrease in 24-hour food intake and water intake, and an increase in the moisture capacity in tongue surface; rats in the Yiguan Jian and TGP groups showed a decrease in cAMP, an increase in cGMP, and a decrease in the cAMP/cGMP, the contents of IL-1, IL-6, and TNF-α were decreased and the content of IL-10 was increased, the percentage of collagen area in the liver was decreased; ALT, AST, γ-GT, ALP, and TBIL levels were decreased in the TGP group, and PI3K, AKT, and mTOR were downregulated at the mRNA and protein levels(P<0.05).Conclusion TGP has a good protective effect on chemical liver injury with pattern of liver yin deficiency in rats. TGP may regulate the balance of yin and yang by inhibiting the PI3K/AKT/mTOR pathway, reducing the secretion of pro-inflammatory cytokines, and increasing the release of anti-inflammatory cytokines.

Objective: To explore the neuroprotective effects of the Shaoyao Gancao decoction (SGD) against excitatory damage in PC12 cells and the role of the Src-NR2-nNOS pathway mediation by SGD in regulating γ-aminobutyric acid (GABA)-glutamate (Glu) homeostasis. Methods: N-Methyl-d-aspartic acid (NMDA) was used to establish a PC12 cell excitability injury model. To investigate the neuroprotective effect of SGD, a cell counting kit-8 (CCK-8) assay was used to determine PC12 cell viability, Annexin V/Propidium Iodide (Annexin V/PI) double staining was used to determine PC12 cell apoptosis, and Ca2+ concentration was observed using laser confocal microscopy. GABA receptor agonists and antagonists were used to analyze the neuroprotective interactions between γ-aminobutyric acid (GABA) and NMDA receptors. Additionally, molecular biology techniques were used to determine mRNA and protein expression in the Src-NR2-nNOS pathway. We analyzed the correlations between the regulatory sites of GABA and NMDA interactions, excitatory neurotoxicity, and brain damage at the molecular level. Results: NMDA excitotoxic injury manifested as a significant decrease in cell activity, increased apoptosis and caspase-3 protein expression, and a significant increase in intracellular Ca2+ concentration. Administration of SGD, a GABAA receptor agonist (muscimol), or a GABAB receptor agonist (baclofen) decreased intracellular Ca2+ concentrations, attenuated apoptosis, and reversed NMDA-induced upregulation of caspase-3, Src, NMDAR2A, NMDAR2B, and nNOS. Unexpectedly, a GABAA receptor antagonist (bicuculline) and a GABAB receptor antagonist (saclofen) failed to significantly increase excitatory neurotoxicity. Conclusions Taken together, these results not only provide an experimental basis for SGD administration in the clinical treatment of central nervous system injury diseases, but also suggest that the Src-NR2A-nNOS pathway may be a valuable target in excitotoxicity treatment.

Objective To establish a human α-synuclein nuclear localization signal transgenic mouse model and investigate the effects of α-synuclein nuclear localization on the behavior of mice.Methods Human α-synuclein nuclear localization signal and EGFP lentiviral vectors were constructed.Transgenic mice were created with the microinjection method.Using PCR and Western Blot method to identify the genotypes and protein expression of the transgenic founder mice and their offsprings.The immunofluorescence was used to examine the localization of human α-synuclein in the mouse brain tissue.The behavioral changes of the transgenic mice were evaluated by the open field test,rotarod test,and O maze test.Results The h SNCA-NLS gene was successfully inserted into the mouse genome,the human α-syn was successfully expressed,and the human α-syn has localized with the nuclear.Further studies found that human α-synuclein nuclear localization signal transgenic mice had significant motor dysfunction,astrocyte proliferation and inflammatory response at 2 months of age and exhibited significant anxiety-like symptoms and reduced expression of the γ-aminobutyric acid(GABA)gene at 9 months of age,which persisted until 12 months of age.Conclusions A human α-synuclein nuclear localization signal transgenic mouse model has been successfully established.The mice exhibit significant motor dysfunction and anxiety-like symptoms.The successful establishment of this model provides a foundation for studying the role of α-syn nuclear localization in Parkinson's disease.

The recognition of the innate immune system is the first line of defense for host protec-tion against invading microbes.Toll-like receptors(TLRs)are well-characterized pattern recogni-tion receptors that are critical for recognizing pathogen-associated molecular pattern(PAMP)and regulating the activation of antigen-presenting cells and key cytokines.Recognition of protozoon parasite genomic DNA by TLR9 activates transcription factors such as NF-κB,IRF3/7,and MAPK,which then induce the production of pro-inflammatory cytokines and type Ⅰ interferons.During parasite infection,MyD88 serves as a critical adapter protein for TLR9 to recognize PAMPs.Here,we review the TLR9-MyD88 pathways and their importance in the inflammatory re-sponse in some common protozoan parasite infections.We also discuss the impact of the subcellular localization of TLR9 in parasitic infectious diseases.By emphasizing current developments in these fields,the review aims to lay the foundation for future research on both the complexity of host-par-asite interactions and the prevention of protozoan parasitic diseases.

Objective:To analyze the plague monitoring results of Marmota sibirica plague natural focus in Hulunbuir Plateau, and to study the characteristics of development and variation of the plague focus. Methods:From 2009 to 2020, monitoring was conducted in Manzhouli City, Prairie Chenbarhu Banner, Xin Barag Left Banner, and Xin Barag Right Banner from May to September each year. The one-day bow-clip method was used to investigate the density of the Spermophilus dauricus, and the 5-meter clamp method was used to investigate other small rodents. The classification and identification of ectoparasite fleas were done by using low power microscope. According to "Diagnostic Criteria for Plague" (WS 279-2008), the detectable materials of rats and fleas were tested, and Yersinia pestis was isolated and cultured. The serums of collected rats were detected by indirect hemagglutination test. Results:From 2009 to 2020, the average density of dauricus in Marmota sibirica plague natural focus of Hulunbuir Plateau was 0.45/hm 2, and the capture rate of other small rodents was 1.25% (787/63 091). A total of 9 species, 8 genera and 4 families of rodents were captured. The total flea infection rate of Spermophilus dauricus was 28.37% (1 945/6 856) and the fleas index was 1.04. A total of 3 species, 3 genera and 2 families of fleas were collected. A total of 6 674 Spermophilus dauricus were dissected and 6 787 fleas were cultured in 2 537 groups. The Yersinia pestis test was negative. A total of 6 795 serum samples from Spermophilus dauricus were detected, of which 11 were positive with a positive detection rate of 0.16%. The highest titer of positive serum was 1∶1 280. Indirect hemagglutination test positive materials were found for 5 consecutive years from 2010 to 2014; the positive serums were found in Manzhouli City and Xin Barag Right Banner. Conclusions:The Yersinia pestis antibady positive serum of Spermophilus dauricus has been detected in Marmota sibirica plague focus of Hulunbuir Plateau for many years, which is worthy of high vigilance. In the future, continuous surveillance should be carried out, bacterial detection should be strengthened, possible plague outbreaks should be detected in time, and various plague prevention and control should be done well.

Organoid models are used to study kidney physiology, such as the assessment of nephrotoxicity and underlying disease processes. Personalized human pluripotent stem cell-derived kidney organoids are ideal models for compound toxicity studies, but there is a need to accelerate basic and translational research in the field. Here, we developed an automated continuous imaging setup with the "read-on-ski" law of control to maximize temporal resolution with minimum culture plate vibration. High-accuracy performance was achieved: organoid screening and imaging were performed at a spatial resolution of 1.1 μm for the entire multi-well plate under 3 min. We used the in-house developed multi-well spinning device and cisplatin-induced nephrotoxicity model to evaluate the toxicity in kidney organoids using this system. The acquired images were processed via machine learning-based classification and segmentation algorithms, and the toxicity in kidney organoids was determined with 95% accuracy. The results obtained by the automated "read-on-ski" imaging device, combined with label-free and non-invasive algorithms for detection, were verified using conventional biological procedures. Taking advantage of the close-to-in vivo-kidney organoid model, this new development opens the door for further application of scaled-up screening using organoids in basic research and drug discovery.
Humans ; Kidney ; Organoids ; Pluripotent Stem Cells

Objective:To evaluate the efficacy of percutaneous curved kyphoplasty (PCKP) for the treatment of osteoporotic vertebral compression fracture (OVCF).Methods:A retrospective cohort study was conducted to analyze the clinical data of 70 patients with OVCF admitted to Third People′s Hospital of Datong from May 2020 to December 2021, including 32 males and 38 females, aged 60-89 years [(73.0±8.7)years]. The patients were treated with PCKP (PCKP group, n=20) or percutaneous vertebroplasty (PVP) (PVP group, n=50). The operation time, intraoperative blood loss, length of hospital stay, excellent rate of bone cement distribution at postoperative 1 day, and leakage rate of bone cement were compared between the two groups. In addition, anterior height ratio of the injured vertebrae, upper and lower Cobb angle, visual analogue scale (VAS), and Oswestry dysfunction index (ODI) were measured preoperatively and at postoperative 24 hours and 3 months. Results:All patients were followed up for 3-4 months [(3.0±0.6)months]. There were no significant differences in the operation time, intraoperative blood loss, and length of hospital stay between the two groups (all P>0.05). At postoperative 1 day, the excellent rate of bone cement distribution was 100% in PCKP group (excellent in 13 patients, good in seven, poor in zero), significantly higher than 82% in PVP group (excellent 21 patients, good in 20, and poor in nine). The leakage rate of bone cement was 0%(0/20) in PCKP group, lower than 20% (10/50) in PVP group ( P<0.05). There were no significant differences in the anterior height ratio of injured vertebrae, upper Cobb angle, lower Cobb angle, VAS and ODI between the two groups before operation (all P>0.05). At postoperative 1 day, PCKP group showed significantly higher anterior height ratio of injured vertebrae and significantly lower upper Cobb angle, lower Cobb angle, VAS, and ODI than those in PVP group ( P<0.05 or 0.01). At postoperative 3 months, PCKP group still showed significantly higher anterior height ratio of injured vertebrae and significantly lower upper Cobb angle and lower Cobb angle than those in PVP group ( P<0.05 or 0.01), but there was no significant difference in VAS and ODI between the two groups (all P>0.05). In PCKP group, the anterior height ratio of the injured vertebrae was significantly increased and the upper Cobb angle, lower Cobb angle, VAS, and ODI index were significantly decreased at postoperative 1 day and 3 months when compared with those before operation (all P<0.05). In PVP group, there were no significant changes in the anterior height ratio of the injured vertebrae, upper Cobb angle, and lower Cobb angle at postoperative 1 day and 3 months when compared with those before operation (all P>0.05), but the VAS and ODI were significantly lowered at postoperative 1 day and 3 months when compared with those before operation (all P<0.05). Conclusion:Compared with PVP, PCKP has better diffusion effect of bone cement in the injured vertebrae and lower incidence of bone cement leakage in the treatment of OVCF, which can effectively promote height recovery of the injured vertebrae, relieve the pain early, and improve spinal function.