OBJECTIVE To study the improvement effects and mechanism of proanthocyanidins （PACs） on steroid-induced osteonecrosis of the femoral head （SONFH） in rabbits based on the receptor-interacting protein kinase 1 （RIPK1）/RIPK3/mixed lineage kinase domain-like protein （MLKL） signaling pathway. METHODS SONFH model in rabbits was induced by injecting Escherichia coli endotoxin+methylprednisolone. The successfully modeled rabbits were randomly divided into Model group （normal saline）， low-dose PACs group （PACs-L group， 11 mg/kg）， high-dose PACs group （PACs-H group， 22 mg/kg）， high-dose PACs+ RIPK1 activator （rRIPK1） group （PACs-H+rRIPK1 group， 22 mg/kg PACs+4 μg/kg rRIPK1）， along with a control group （normal saline）， with 6 rabbits in each group. Each administration group was given relevant medicine once a day intragastrically/via injection， for 4 consecutive weeks. After the last administration， the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in rabbit serum were measured. The changes in the microstructure of rabbit femurs， including bone mineral density （BMD）， trabecular thickness （Tb.Th）， trabecular number （Tb.N）， and trabecular separation （Tb. Sp） were examined. The histopathological features of rabbit femoral tissues were observed， and the apoptotic status of cells within the rabbit femoral tissues was detected. The mRNA expressions of vascular endothelial growth factor （VEGF） and bone morphogenetic protein 2 （BMP2） in rabbit femoral tissues were determined. The expressions of RIPK1/RIPK3/MLKL signaling pathway-related proteins in femoral tissues were detected. RESULTS Compared with the Control group， serum contents of TNF-α and IL-6， Tb.Sp， empty bone cavity rate， cell apoptosis rate， phosphorylation levels of RIPK1， RIPK3 and MLKL in femoral tissue were significantly increased in the Model group （P＜0.05）. BMD， Tb.Th， Tb.N， as well as the mRNA expression of VEGF and BMP2， along with protein expression of caspase-8， in the femoral tissues were all decreased （P＜0.05）. The bone cells in the femoral tissue were unevenly distributed， and the trabeculae were arranged sparsely. Compared with the Model group， the aforementioned quantitative indicators （P＜0.05） and pathological changes in all dosage groups of PACs showed significant improvements. Compared with the PACs-H group， the aforementioned quantitative indicators （P＜0.05） and pathological changes in the PACs-H+rRIPK1 group showed significant reversal. CONCLUSIONS PACs can ameliorate SONFH in rabbits， and its mechanism of action may be related to the inhibition of the activation of the RIPK1/RIPK3/MLKL signaling pathway， suppression of apoptosis in femoral tissue cells， and promotion of angiogenesis.

Objective To investigate the mechanism by which Scutellaria barbata D.Don water extract(SBW)inhibits lung metastasis of breast cancer by regulating the immune microenvironment.Methods A mouse model of lung metastasis was established using 4T1 breast cancer cells.Mice were divided into a control group(n=6)and an SBW treatment group(n=6).The effect of SBW on tumor growth was assessed by measuring the volume of the primary tumor,and the inhibitory effect on lung metastasis was evaluated by observing the number and area of metastatic nodules in lung tissue using H&E staining.Flow cytometry was used to analyze changes in the composition of immune cells in the tumor,peripheral blood,and lung tissue.Results Compared with the control group,the SBW treatment group inhibited the growth of the primary tumor(P<0.01)and reduced the number and area of lung metastatic nodules(P<0.01).Flow cytometry analysis showed that after SBW treatment,the numbers of CD86+macrophages(P<0.001)and polymorphonuclear myeloid-derived suppressor cells(PMN-MDSCs)(P<0.05)in the tumor tissue were increased.In lung tissue,the numbers of CD86+macrophages,natural killer T(NKT)cells,natural killer(NK)cells,and PMN-MDSCs were also elevated(P<0.05).Meanwhile,the numbers of regulatory T cells(Tregs)(P<0.05),CD206+macrophages(P<0.01),and monocytic myeloid-derived suppressor cells(M-MDSCs)(P<0.05)in both tumor and lung tissues were decreased.Conclusion SBW inhibits the breast cancer growth and lung metastasis by regulating the recruitment and distribution of immune cells in the tumor,peripheral blood,and lung tissue,thereby enhancing anti-tumor immune responses and reducing immune suppression.

Objective:To investigate the effects of subcutaneous immunotherapy (SCIT) on patients′ immune markers and metabolic levels in the early stage of allergen treatment, and to gain insight into the role of SCIT in regulating immune responses and metabolic levels, so as to provide reference data for the further discovery of potential biomarkers.Methods:A longitudinal study was used to include 40 subjects who underwent SCIT with dust mite allergens in the Department of Pediatrics of the First Affiliated Hospital of Guangzhou Medical University between November 2017 and February 2022, including 20 subjects each of single mite subcutaneous immunotherapy (SM-SCIT) and double mite subcutaneous immunotherapy (DM-SCIT). In this study, levels of dust mite allergen-specific antibodies and polyunsaturated fatty acid metabolism were measured before and 12 months after treatment, while pulmonary function tests were performed. The therapeutic effects of the patients were followed up by visual analogue scale (VAS), asthma control test (ACT) and total medication scores (TMS). The results were statistically analyzed using t-test and Mann-Whitney U-test. Results:After 12 months of treatment with SCIT, both groups showed a significant decrease in total VAS score (SM-SCIT: Z=-2.298, P<0.05; DM-SCIT: Z=-3.411, P<0.001); total ACT score (SM-SCIT: Z=-2.054, P<0.05; DM-SCIT: Z=-2.014, P<0.05) and total medication scores (SM-SCIT: Z=-3.799, P<0.000 1; DM-SCIT: Z=-3.474, P<0.001) were significantly higher, in addition to significantly higher MMEF75/25 values in the DM-SCIT group ( t=-2.253, P<0.05). There was no significant change in sIgE in the SM-SCIT group ( P>0.05), and the sIgG4 levels of the Der p, Der f, p 1, p 2, f 2, and p 21 fractions were significantly elevated ( Z=-2.651, -3.771, -2.949, -2.912, -2.725, -2.128, and -3.285, respectively, all P<0.05); The sIgE of Der p 2, f 2, p 7 and p 23 fractions( Z=-2.651, -3.771, -2.949, -2.912, -2.725, -2.128, -3.285, all P<0.05) and the sIgG4 levels of the Der p, Der f, p 1, p 2, f 1, f 2, p 10, p 21 and p 23 fractions ( Z=-3.808, -3.845, -3.061, -2.688, -2.464, -3.211, -2.371, -2.091, -2.427, all P<0.05) of the DM-SCIT group were significantly elevated. Metabolomics analysis showed that arachidonic acid, docosahexaenoic acid, docosapentaenoic acid, eicosapentaenoic acid, 5, 9, 12-octadecatrienoic acid, 5(S)-hydroxylated eicosatetraenoic acid, and dihomo-gamma-linolenic acid were significantly elevated at the beginning of the treatment period after SM-SCIT treatment ( Z of -2.191, -2.497, -1.988, -2.090, -2.19, -2.803, -2.073, all P<0.05); 5(S)-hydroxylated eicosatetraenoic acid showed elevated and alpha-linolenic acid, eicosadienoic acid, and eicosapentaenoic acid were significantly decreased in the DM-SCIT group after treatment ( Z=-1.988, -2.090, -2.497, -1.988, respectively, all P<0.05). Correlation analysis showed that arachidonic acid was significantly negatively correlated with changes in dust mite-specific IgG4 ( r=-0.499, P<0.05), and that alpha-linolenic acid, 5, 9, 12-octadecatrienoic acid, and eicosapentaenoic acid were positively correlated with the ΔsIgG4 of the dust mite der p 2 ( r=0.451, 0.420, 0.474, respectively; all P<0.05). Conclusion:Significant changes in allergen-specific antibody levels and polyunsaturated fatty acid metabolism levels occur during SCIT, and the two may interact and influence each other.

Objective:To investigate the effects of subcutaneous immunotherapy (SCIT) on patients′ immune markers and metabolic levels in the early stage of allergen treatment, and to gain insight into the role of SCIT in regulating immune responses and metabolic levels, so as to provide reference data for the further discovery of potential biomarkers.Methods:A longitudinal study was used to include 40 subjects who underwent SCIT with dust mite allergens in the Department of Pediatrics of the First Affiliated Hospital of Guangzhou Medical University between November 2017 and February 2022, including 20 subjects each of single mite subcutaneous immunotherapy (SM-SCIT) and double mite subcutaneous immunotherapy (DM-SCIT). In this study, levels of dust mite allergen-specific antibodies and polyunsaturated fatty acid metabolism were measured before and 12 months after treatment, while pulmonary function tests were performed. The therapeutic effects of the patients were followed up by visual analogue scale (VAS), asthma control test (ACT) and total medication scores (TMS). The results were statistically analyzed using t-test and Mann-Whitney U-test. Results:After 12 months of treatment with SCIT, both groups showed a significant decrease in total VAS score (SM-SCIT: Z=-2.298, P<0.05; DM-SCIT: Z=-3.411, P<0.001); total ACT score (SM-SCIT: Z=-2.054, P<0.05; DM-SCIT: Z=-2.014, P<0.05) and total medication scores (SM-SCIT: Z=-3.799, P<0.000 1; DM-SCIT: Z=-3.474, P<0.001) were significantly higher, in addition to significantly higher MMEF75/25 values in the DM-SCIT group ( t=-2.253, P<0.05). There was no significant change in sIgE in the SM-SCIT group ( P>0.05), and the sIgG4 levels of the Der p, Der f, p 1, p 2, f 2, and p 21 fractions were significantly elevated ( Z=-2.651, -3.771, -2.949, -2.912, -2.725, -2.128, and -3.285, respectively, all P<0.05); The sIgE of Der p 2, f 2, p 7 and p 23 fractions( Z=-2.651, -3.771, -2.949, -2.912, -2.725, -2.128, -3.285, all P<0.05) and the sIgG4 levels of the Der p, Der f, p 1, p 2, f 1, f 2, p 10, p 21 and p 23 fractions ( Z=-3.808, -3.845, -3.061, -2.688, -2.464, -3.211, -2.371, -2.091, -2.427, all P<0.05) of the DM-SCIT group were significantly elevated. Metabolomics analysis showed that arachidonic acid, docosahexaenoic acid, docosapentaenoic acid, eicosapentaenoic acid, 5, 9, 12-octadecatrienoic acid, 5(S)-hydroxylated eicosatetraenoic acid, and dihomo-gamma-linolenic acid were significantly elevated at the beginning of the treatment period after SM-SCIT treatment ( Z of -2.191, -2.497, -1.988, -2.090, -2.19, -2.803, -2.073, all P<0.05); 5(S)-hydroxylated eicosatetraenoic acid showed elevated and alpha-linolenic acid, eicosadienoic acid, and eicosapentaenoic acid were significantly decreased in the DM-SCIT group after treatment ( Z=-1.988, -2.090, -2.497, -1.988, respectively, all P<0.05). Correlation analysis showed that arachidonic acid was significantly negatively correlated with changes in dust mite-specific IgG4 ( r=-0.499, P<0.05), and that alpha-linolenic acid, 5, 9, 12-octadecatrienoic acid, and eicosapentaenoic acid were positively correlated with the ΔsIgG4 of the dust mite der p 2 ( r=0.451, 0.420, 0.474, respectively; all P<0.05). Conclusion:Significant changes in allergen-specific antibody levels and polyunsaturated fatty acid metabolism levels occur during SCIT, and the two may interact and influence each other.

BACKGROUND:Previous brain studies have mostly focused on adults and fetuses,and the developmental characteristics of young children's brainstems have rarely been studied. OBJECTIVE:To observe the brainstem development characteristics of healthy young children and to explore the age-related differences and their correlation with sex. METHODS:From January 2019 to April 2022,a retrospective study of 3.0T MRI images of 174 children aged 2 to 6 years in the Affiliated Hospital of Inner Mongolia Medical University was conducted,and the median sagittal diameter,area and angle of the brainstem(including midbrain,pons and medulla oblongata)were measured. RESULTS AND CONCLUSION:There is an age-related increase in the anterior and posterior diameters of the midbrain,pons and medulla oblongata in the 2-5 years old group as well as in the longitudinal diameter and area of the midbrain,pons and medulla oblongata in the 2-6 years old group.Except for the longitudinal diameter of the medulla oblongata,all others show a positive correlation with age(r>0,P<0.05).In the 2-3 years old group and 4-5 years old group,the children are in the rapid growth and development stage,and these two age groups can be used as the key observation indicators for the development of young children.The anterior-posterior diameter,longitudinal diameter,area of the pons and total brainstem area are strongly correlated with age,which can be used as the key observation indicators for the brainstem development in young children.

BACKGROUND:Nowadays,there are increasing reports on the digitization and visualization system of acupuncture points for adults in traditional Chinese medicine,and the digitization and visualization system of acupuncture points for children in traditional Chinese medicine and the simulation system of acupuncture manipulation for Mongolian medicine training have been reported.However,there are no reports on relevant systems for children in Mongolian medicine. OBJECTIVE:To develop a simulation system of acupuncture points for children in Mongolian medicine,in the hope that it can be used for clinical teaching,manipulation practice and research on acupuncture safety. METHODS:Based on the tomographic anatomical dataset of preschool boys,a three-dimensional(3D)digital virtual anatomical model of children with multiple internal organs and tissues was constructed by using PhotoShop.2021 and Digihuman Reconstruction System software.The relevant annotation information database of 27 acupoints such as Dinghui acupoint of Mongolian medicine was compiled by the Unity database language.The Mongolian gold needle and silver needle were selected to record the acupuncture point teaching video on the 3D printed head and neck resin model of children.In Unity3D software,children's anatomical model,acupoint annotation information database and acupuncture operation video were integrated and coded,and a 3D digital children's Mongolian acupuncture acupoint visualization system integrating simulation acupuncture training,clinical teaching and acupuncture safety research was successfully created. RESULTS AND CONCLUSION:This study was based on real children's specimens.In order to reduce the error of two-dimensional segmentation,the manual layer-by-layer segmentation section image method was used to ensure the accuracy of the 3D model to the greatest extent.The Digihuman Reconstruction System was used to extract and save the individual segmentation data while maximizing the accuracy of the 3D model.PhotoShop.2021 software was used to complete the 3D reconstruction of the outer skin of the head and neck of children and the internal bony structure,cervical spinal cord,blood vessels and nerves,muscles and ligaments.After 3D reconstruction,the basic morphology of each independent structure and the integrity of the overall contour were verified in MeshLab software and the final fine adjustment and anatomical position confirmation were conducted using 3-matic research 13.0 software.The real anatomical morphology of the head and neck of preschool children was successfully simulated and restored.Unity3D software was used to integrate the 3D model of children,acupuncture operation video and acupoint annotation database,and the 3D digital Mongolian acupuncture acupoint visualization system for children was successfully constructed.Based on the real continuous fault ultra-thin dataset of preschool boys in China,China's first 3D digitization and visualization system of acupuncture points in the head and neck of children in Mongolian medicine was developed.Compared with the previous acupuncture soft works,this system is more suitable for the anatomical morphological development characteristics of Asian children,and has a high value of application in the fields of research on the safety of acupuncture in Mongolian medicine,clinical teaching and acupuncture simulation training.

BACKGROUND:The calcaneus is located in the lower posterior part of the foot and is heavily stressed.Calcaneus is complex and irregular in shape,surrounded by thin cortical bone and filled with a large number of trabeculae.The study of the microstructure,movement,and distribution of trabeculae is helpful to improve the understanding of calcaneus fracture. OBJECTIVE:The image data of calcaneus were obtained by scanning the calcaneus specimens with micro-computed tomography,and the structure of trabecular bone in calcaneus was analyzed to explore the morphology,distribution and structural characteristics of trabecular bone in calcaneus. METHODS:Dry adult calcaneus specimens were continuously scanned by micro-computed tomography,and the images were obtained after scanning and stored in DICOM format.Image data were imported into Hiscan Analyzer software to display clear and complete images of the sagittal plane,coronal plane,and the horizontal plane of the adult calcaneus.The trabecular movement of bone was observed layer by layer.According to the trabecular movement characteristics,the sagittal plane of the calcaneus was divided into six parts.A 49-mm2 region of interest was selected for each part at the same thickness as 7 mm.The three-dimensional microstructure of calcaneus and trabecular bone was obtained after three-dimensional reconstruction.After binarization,the volume fraction,surface density,trabecular thickness,trabecular space,and trabecular number parameters of the trabecular bone in the region of interest were calculated by software. RESULTS AND CONCLUSION:(1)The cortical layer of the calcaneus was very thin and filled with a large amount of cancellous bone,and the cortical layer of the horn of Gissane was obviously thickened.(2)The trabecular volume fraction in the upper part of the calcaneus was greater than that in the anterior part of the lower part of the calcaneus,the central triangle,the posterior part of the lower part of the calcaneus,and the base of the calcaneus,and the trabecular volume fraction in the tubercle of the calcaneus was greater than that in the anterior part of the lower part of the calcaneus,the central triangle,the posterior part of the lower part of the calcaneus,and the base of the calcaneus.The surface density of the trabecular bone in the tubercle of the calcaneus was higher than that in the front of the lower calcaneus,the middle triangle area,and the bottom of calcaneus,and the surface density of the trabecular bone in the upper part of calcaneal bone,and the lower part of the calcaneus was higher than that in the middle triangle area.The thickness of the trabecular bone in the upper part of the calcaneus was greater than that in the tubercle of the calcaneal bone.The bone trabecular space in the middle triangle was larger than that in the upper part of the calcaneus and calcaneal tubercles.The number of bone trabeculae in the calcaneal tubercles was greater than that in the middle triangle area.(3)These results indicate that the trabeculae of rod bone were mainly distributed in the middle triangle area.The surface density of trabeculae was the smallest,the volume fraction was smaller,and the space between trabeculae was the largest.This part of the bone is relatively loose.The compression resistance is poor when subjected to high impact.The trabecular bone fractures first occur,which is a prone site for fractures.

Objective To evaluate the efficacy of dexmedetomidine in preventing postoperative de-lirium in the patients with schizophrenia. Methods Ninety patients with schizophrenia of both sexes, with American Society of Anesthesiologists physical status Ⅰ or Ⅱ, aged 20-60 yr, weighing 45-90 kg, with a history of long-term use of antipsychotics, scheduled for elective emergency operation under general anes-thesia, were divided into 3 groups ( n=30 each) using a random number table method: high-dose dexme-detomidine group ( group HD ) , low-dose dexmedetomidine group ( group LD ) and control group ( group C) . In HD and LD groups, dexmedetomidine was intravenously injected in doses of 1. 0 and 0. 5 μg∕kg, respectively, over 10 min prior to induction of anesthesia, followed by continuous infusion at 0. 4 and 0. 2μg · kg-1 · h-1 , respectively, until the end of operation. Anesthesia was maintained with propofol, remifentanil and sevoflurane, and bispectral index value was maintained at 40-55. Patient-controlled intra-venous analgesia was performed within 48 h after operation, and visual analogue scale score was main-tained≤3. The end-tidal concentration of sevoflurane ( ETsev) was recorded at 30 min after endotracheal intubation. The sleep quality was evaluated and scored at 6 h and 1, 2 and 3 days after operation. The oc-currence of delirium during emergency from anesthesia and within 3 days after operation was recorded. The occurrence of postoperative adverse reactions such as bradycardia, hypotension and hypoxemia was also re-corded. Results Compared with group C, the ETsev, sleep quality scores at each time point after opera-tion and incidence of delirium during emergency from anesthesia and within 3 days after operation ( 3％) were significantly decreased in group HD, and the ETsev, sleep quality scores at 6 h after operation and in-cidence of delirium during emergency from anesthesia were significantly decreased ( P<0. 05) , and no sig-nificant change was found in the incidence of delirium within 3 days after operation in group LD ( P>0. 05) . The ETsev and incidence of delirium within 3 days after operation were significantly lower in group HD than in group LD ( P<0. 05) . There was no significant difference in the incidence of bradycardia, hypotension or hyoxemia among C, HD and LD groups (P>0. 05). Conclusion Dexmedetomidine given as a loading dose of 1. 0μg∕kg followed by a maintenance dose of 0. 4μg·kg-1 ·h-1 can prevent postoperative delirium effectively in the patients with schizophrenia.

Objective To evaluate the effect of dexmedetomidine on Toll-like receptor 4 ( TLR4)/myeloid differentiation factor 88 ( MyD88)/nuclear factor Kappa B ( NF-κB) signaling pathway in the pe-ripheral blood mononuclear cells of elderly patients with diabetes mellitus undergoing lower extremity surger-y. Methods Forty elderly patients of both sexes with type 2 diabetes mellitus, aged 65-80 yr, with body mass index of 18. 5-27. 9 kg/m2 , of American Society of Anesthesiologists physical status Ⅱ or Ⅲ, with New York Heart Association Ⅰ or Ⅱ, undergoing lower extremity surgery with tourniquets under general anesthesia, were divided into 2 groups ( n=20 each) by using a random number table method: control group ( group C) and dexmedetomidine group ( group D) . Combined intravenous-inhalational anesthesia was applied. Dexmedetomidine was infused over 15 min in a dose of 1μg/kg after induction of anesthesia, fol-lowed by a continuous infusion of 0. 5 μg·kg-1 ·h-1 until the end of surgery in group D, while the equal volume of normal saline was given instead of dexmedetomidine in group C. Before using the the tourniquet and at 15 min, 1 h and 24 h after loosing the tourniquet, arterial blood samples were collected for determi-nation of the expression of TLR4, NF-κB and MyD88 in peripheral blood mononuclear cells and concentra-tions of tumor necrosis factor-alpha ( TNF-α) , interleukin-1β ( IL-1β) , cardiac troponin I ( cTnI ) and creatine kinase-MB ( CK-MB) in plasma. Results Compared with group C, the expression of TLR4, NF-κB and MyD88 in peripheral blood mononuclear cells was significantly down-regulated, and the concentra-tions of TNF-α, IL-1β, cTnI and CK-MB in plasma were decreased at each time point after loosing the tourniquet in group D ( P＜0. 05) . Conclusion The mechanism by which dexmedetomidine reduces myo-cardial damage may be related to inhibiting TLR4/MyD88/NF-κB signaling pathway and reducing systemic inflammatory responses in elderly patients with diabetes mellitus undergoing lower extremity surgery.

Objective To select a simple, stable and reliable mouse model of alcoholic liver disease. Methods The mouse models of alcoholic liver disease were induced by oral gavage ethanol or Lierber?DeCarli ethanol liquid diet for 8 weeks. The food intake and body weight were recorded. Pathological changes were examined using HE staining. Liver injury was assessed by the activities of serum ALT, AST, AKP and γ?GT, and serum and hepatic TC and TG. Results After modeling, both models showed significantly increased activities of serum ALT, AST, AKP, and contents of serum and hepatic TG (P<0?05), indicating the successful development of alcoholic steatohepatitis. However, oral ethanol gavage led to body weight loss and weak mental state. Ethanol liquid diet less affected the body weight and mental state. Ethanol liquid diet enhanced liver to?body weight ratio and serum TC, but oral gavage of ethanol did not. The changes of serum ALT, AST, serum and hepatic TG, and hepatic steatosis in the ethanol liquid diet models were more severe than those in the oral gavage ethanol models, suggesting that Lierber?DeCarli ethanol liquid diet led to more serious liver injury than oral gavage ethanol. Conclusions Lierber?DeCarli ethanol liquid diet model is better than oral gavage ethanol model, and is more suitable for studies on mechanisms and evaluation of hepato?protective drugs for alcoholic liver disease.