ObjectiveTo investigate the therapeutic effect of Siegesbeckiae Herba water decoction （SWD） at different doses on atherosclerosis （AS） in a mouse model induced by a high-fat diet and analyze its potential mechanism of action. MethodsThirty-six male ApoE^-/- mice were randomly divided into six groups： blank control group， model group， low-dose， medium-dose， and high-dose SWD groups， and positive control group. Firstly， the AS mouse model was created by feeding mice a high-fat diet. After successful modeling， the low-， medium-， and high-dose SWD groups were intragastrically administered with SWD at 0.65， 1.3， 2.6 g·kg^-1， respectively. The positive control group was intragastrically administered with 30 mg·kg^-1 of atorvastatin calcium aqueous solution， while the blank and model groups received an equal volume of 0.9% sodium chloride solution via oral gavage， all administered for 12 weeks. During the administration period， the general condition of the mice was observed and recorded daily. Before sampling， color Doppler ultrasound was performed to observe the pathological changes in atherosclerotic plaques in the aortic wall of mice. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in aortic tissue in mice， and oil red O staining was used to detect the atherosclerotic plaque area in the aorta. Enzyme-linked immunosorbent assay （ELISA） was used to detect the serum lipid indices and the levels of interleukins （IL-1β， IL-4， IL-6， and IL-10） and tumor necrosis factor-α （TNF-α） in mice. Protein expression levels of IKKα， IKKβ， and NF-κB p65 in mouse aortic tissue were detected by Western blot. ResultsCompared with the blank control group， the model group showed a significant increase in body weight. The results of color Doppler ultrasound showed enhanced vascular wall echo， suggesting the presence of atherosclerotic plaques. HE staining showed foam cell aggregation， fibrous connective tissue proliferation， and vascular intima injury in the aortic tissue. Oil red O staining showed a significant increase in the plaque area in the aortic tissue （P<0.01）. ELISA results indicated significantly elevated levels of IL-1β， IL-6， TNF-α， total cholesterol （TC）， triglyceride （TG）， and low-density lipoprotein （LDL） in mouse serum （P<0.01）， as well as significantly decreased levels of IL-4， IL-10， and high-density lipoprotein （HDL） （P<0.01）. Western blot results showed that the expression of IKKα， IKKβ， and NF-κB p65 in mouse aortic tissue increased significantly （P<0.01）. Compared with those in the model group， mice in the middle- and high-dose SWD groups showed significant weight loss. In the high-dose group， the aortic vascular wall echoes were weakened， and the atherosclerotic plaques were reduced. The aortic lesions of mice in the medium- and high-dose SWD groups were significantly alleviated. The plaque area percentage showed an inverse correlation with the administered dose in all groups treated with SWD （P<0.05）. In the medium-dose SWD group， serum levels of IL-1β， IL-6， TNF-α， TC， TG， and LDL were significantly decreased （P<0.05， P<0.01）， while those of IL-4 and IL-10 were significantly increased （P<0.01）. In the high-dose SWD group， levels of IL-1β， IL-6， TNF-α， TC， TG， and LDL were significantly decreased （P<0.01）， while IL-4， IL-10， and HDL were significantly increased （P<0.01）. The IKKα and IKKβ expression was significantly decreased in the low-dose SWD group （P<0.05）， and IKKα， IKKβ， and NF-κB p65 were significantly decreased in the medium- and high-dose SWD groups （P<0.05， P<0.01）. ConclusionSWD may exert therapeutic effects on AS by regulating the expression of related inflammatory factors through the NF-κB signaling pathway， thereby reducing inflammation， plaque area， and lipid content in the body.

There are millions of patients with taxane-induced peripheral neuropathy （TIPN）， and there is no effective treatment or prevention measure in clinical practice. The occurrence of TIPN may be related to the dosage form of paclitaxel drugs， genetic and molecular markers， drug dosage and chemotherapy cycle， patient factors， etc. At present， drugs for treating TIPN mainly include those that inhibit axonal degeneration （such as dosazosin， tamsulosin）， prevent mitochondrial dysfunction （such as glutathione trisulfides， antioxidants α -lipoic acid）， improve calcium imbalance in the internal environment （Shaoyao gancao decoction， N-type voltage-gated calcium channel inhibitor IPPQ）， and inhibit neuroinflammation （such as chemokine inhibitors and selective interleukin-8 receptor inhibitors DF2726A）. Further exploration of drug treatment strategies targeting different induction mechanisms is expected to become a new direction for precise clinical prevention and personalized treatment of TIPN.

BACKGROUND:Intervertebral disc degeneration is clinically considered to be the main cause of low back pain,but due to the unclear pathogenesis of intervertebral disc degeneration,there is still a lack of effective means to delay the progression of the disease.Single-cell RNA sequencing technology can amplify and sequence mRNA at the single-cell level,reveal the gene expression intensity of a single cell,discover different cell subsets in tissues according to the heterogeneity of cells,study the pathogenesis of intervertebral disc degeneration at the molecular level,and provide a new theoretical basis for its early diagnosis and treatment. OBJECTIVE:To introduce the basic principles of single-cell RNA sequencing technology and review the research progress of single-cell RNA sequencing technology in intervertebral disc degeneration in recent years. METHODS:A computer was used to search PubMed,Web of Science,CNKI and WanFang databases for the literature published from 2012 to 2022.Key words were"single-cell RNA sequencing,intervertebral disc degeneration,sequencing Technology"in Chinese and English.Duplicate,poor-quality and irrelevant articles were excluded;a total of 70 articles were eventually included. RESULTS AND CONCLUSION:(1)We identified new cell subsets such as homeostatic chondrocytes,hypertrophy chondrocyte-like nucleus pulposus cells and fibrous nucleus pulposus cells,identified the marker genes and transcription factors of these cell subsets,and described the functions,differentiation paths and cell fate of these cell subsets during the development and progression of intervertebral disc degeneration,and proposed the concept of progenitor nucleus pulposus cells.A cell subpopulation with progenitor nucleus pulposus cells properties was identified and its effectiveness in treating intervertebral disc degeneration was verified in mice.(2)Fibro chondrocyte-like annulus fibrosus cells and annulus fibrosus stem cells with both cartilage and fiber properties were identified,and a new type of composite hydrogel was prepared by combining fibrous cartilage inducers silk fibroin and hyaluronic acid in vitro.Experiments in mice demonstrated that this hydrogel could repair both annulus fibrosus tissue and cartilage matrix,and was remarkably effective in the treatment of intervertebral disc degeneration.(3)Regulatory chondrocytes were found in endplate cartilage.Two distinct fates in the progression of intervertebral disc degeneration were analyzed and the differential genes in the two fates were identified.Intercellular communication analysis indicated that regulatory chondrocytes interact with endothelial cells to promote angiogenesis.(4)Immune cells such as macrophages,T cells,myeloid progenitor cells and neutrophils were identified in the degenerated intervertebral disc tissues,demonstrating the existence of immune response during intervertebral disc degeneration.It was found that apolipoprotein induced the polarization of macrophages M1 and M2 subtypes,and this polarization process affected the activity of progenitor nucleus pulposus cells by amplifying the inflammatory response through the MIF signaling pathway.

Objective To explore the difference in efficacy of metoprolol versus ivabradine in the treatment of postural orthostatic tachycardia syndrome(POTS)in the elderly after COVID-19 infection.Methods A total of 110 patients diagnosed with POTS at our department from Decem-ber 1,2022 to January 31,2023 were included.According to their drug regimen,they were divided into metoprolol group(62 patients)and ivabradine group(48 patients).On the 28th day of out-patient follow-up,the resting heart rate,heart rate of 10 min of standing,symptom disappearance rate,hospitalization rate,and mortality rate were compared between the two groups.Results On the 28th day of treatment,the resting heart rate and postural heart rate for 10 min were decreased in both groups when compared with the levels at initial diagnosis(P＜0.01).And there were no significant differences in the two types of heart rate between the two groups on the 28th day(71.0±7.0 vs 72.1±7.0,P=0.401;76.5±7.2 vs 77.4±7.6,P=0.573).No obvious differences were observed between the two groups in symptom disappearance rate,hospitalization rate,or mortality rate(88.7％vs 89.6％,3.2％vs2.1％,0％vs 0％,P＞0.05).Conclusion Metoprolol and ivabradine can effectively treat POTS in the elderly patients after COVID-19 infection.

BACKGROUND:Tissue engineering has brought new hope to the clinical challenge of liver failure,and the preparation of plant-derived decellularized fiber scaffolds holds significant importance in liver tissue engineering. OBJECTIVE:To prepare apple tissue decellularized scaffold material by using fresh apple slices and a solution of sodium dodecyl sulfate,and assess its biocompatibility. METHODS:Fresh apples were subjected to decellularization using phosphate buffer saline and sodium dodecyl sulfate solution,separately.Afterwards,the decellularized apple tissues and apple decellularized scaffold materials were decontaminated with phosphate buffer saline.Subsequently,scanning electron microscopy was used to assess the effectiveness of decellularization of the apple materials.Adipose-derived mesenchymal stem cells were extracted from the inguinal fat BALB/C of mice,and their expression of stem cell-related markers(CD45,CD34,CD73,CD90,and CD105)was identified through flow cytometry.The cells were then divided into a scaffold-free control group and a scaffold group.Equal amounts of adipose-derived mesenchymal stem cells were seeded onto both groups.The biocompatibility of the decellularized scaffold with adipose-derived mesenchymal stem cells was evaluated using CCK-8 assay,hematoxylin-eosin staining,and phalloidine staining.Cell adhesion and growth on the scaffold were observed under light microscopy and scanning electron microscopy.Furthermore,the scaffold was subdivided into the non-induced group and the hepatogenic-induced group.Adipose-derived mesenchymal stem cells were cultured on the decellularized apple scaffold,and they were cultured for 14 days in regular culture medium or hepatogenic induction medium for comparison.Immunofluorescent staining using liver cell markers,including albumin,cytokeratin 18,and CYP1A1,was performed.Enzyme-linked immunosorbent assay was used to detect the secretion of alpha fetoprotein and albumin.Additionally,scanning electron microscopy was employed to observe the morphology of the induced cells on the scaffold,verifying the expression of liver cell-related genes on the decellularized scaffold material.Finally,the cobalt-60 irradiated and sterilized decellularized apple scaffolds were transplanted onto the surface of mouse liver and the degradation of the scaffold was observed by gross observation and hematoxylin-eosin staining after 28 days. RESULTS AND CONCLUSION:(1)The scanning electron microscopy results revealed that the decellularized apple scaffold material retained a porous structure of approximately 100 μm in size,with no residual cells observed.(2)Through flow cytometry analysis,the cultured cells were identified as adipose-derived mesenchymal stem cells.(3)CCK-8 assay results demonstrated that the prepared decellularized apple tissue scaffold material exhibited no cytotoxicity.Hematoxylin-eosin staining and phalloidine staining showed that adipose-derived mesenchymal stem cells were capable of adhering and proliferating on the decellularized apple tissue scaffold.(4)The results obtained from immunofluorescence staining and enzyme-linked immunosorbent assay revealed that adipose-derived mesenchymal stem cells cultured on the decellularized apple scaffolds exhibited elevated expression of liver-specific proteins,including albumin,alpha-fetoprotein,cytokeratin 18,and CYP1A1.These results suggested that they were induced differentiation into hepatocyte-like cells possessing functional characteristics of liver cells.(5)The decellularized apple scaffold implanted at 7 days has integrated with the liver,with partial degradation of the scaffold observed.By 28 days,the decellularized apple scaffold has completely degraded and has been replaced by newly-formed tissue.(6)The results indicate that the decellularized scaffold material derived from apple tissue demonstrates favorable biocompatibility,promoting the proliferation,adhesion,and hepatic differentiation of adipose-derived mesenchymal stem cells.

Objective:To analyze the clinical characteristics, curative effect related factors and follow-up situation of bilateral sudden sensorineural hearing loss (BSSHL).Methods:The clinical data of 169 patients(338 ears) with BSSHL were retrospectively summarized, and the demographic characteristics, predisposing factors, concomitant symptoms and diseases, and audiological characteristics were statistically described. Additionally, influencing factors of curative effect and prognosis were statistically analyzed.Results:Among the 169 patients, 50.9% (86/169) of patients had at least one incentive, with cold and fatigue being the most common incentives(both 23/169). There were high rates of accompanying symptoms including tinnitus (150/169, 88.8%) and dizziness (100/169, 59.2%). Hypertension(49/169, 29.0%)and diabetes(23/169, 13.6%)were the most common concomitant diseases observed. Most cases exhibited all frequencies involvement, with flat type and total deafness type accounting for 83.1%(281/338 ears). The most common degree of hearing loss was total deafness(86/338 ears), with approximately 60.1%(203/338 ears) of the cases being severe or worse. The total effective rate of treatment was only 29.0%, but it increased to 39.5% for patients with course of disease≤14 days, however, when course of disease>30 days, the effective rate decreased sharply to 3.6%, showing a significant difference between these two groups( χ2=13.776,<0.01). Different types of hearing curves showed statistically significant difference in efficiency( χ2=14.782, P<0.01). Comparing the hearing improvements of 28 BSSHL patients from admission to discharge and from discharge to follow-up, it was found that the hearing improvement of the two groups showed statistically significant difference at the frequencies of L-250 Hz, L-500 Hz, R-125 Hz, R-250 Hz and R-500 Hz( Z value was -2.495, -3.083, -3.970, -3.388 and -3.264 respectively, all P<0.05). The proportion of patients with elevated serum IgE was much higher than that of the normal population. Conclusion:BSSHL patients suffer from serious hearing loss, and many also experience tinnitus and vertigo symptoms. Due to the poor efficiency of treatment, it is better for patients to be treated within 30 days of onset. For patients of hearing loss in the low frequency range, hearing improvement is more significant during hospitalization. And the occurrence of BSSHL may involve an immune mechanism.

Objective To explore the mechanism by which soybean isoflavone(SI)reduces calcium overload induced by cerebral ischemia-reperfusion(I/R).Methods Forty-eight SD rats were randomized into 4 groups to receive sham operation,cerebral middle artery occlusion for 2 h followed by 24 h of reperfusion(I/R model group),or injection of adeno-associated virus carrying Frizzled-2 siRNA or empty viral vector into the lateral cerebral ventricle after modeling.Western blotting was used to examine Frizzled-2 knockdown efficiency and changes in protein expressions in the Wnt/Ca2+signaling pathway.Calcium levels and pathological changes in the ischemic penumbra(IP)were measured using calcium chromogenic assay and HE staining,respectively.Another 72 SD randomly allocated for sham operation,I/R modeling,or soy isoflavones pretreatment before modeling were examined for regional cerebral blood flow using a Doppler flowmeter,and the cerebral infarct volume was assessed using TTC staining.Pathologies in the IP area were evaluated using HE and Nissl staining,and ROS level,Ca2+level,cell apoptosis,and intracellular calcium concentration were analyzed using immunofluorescence assay or flow cytometry;the protein expressions of Wnt5a,Frizzled-2,and P-CaMK II in the IP were detected with Western blotting and immunohistochemistry.Results In rats with cerebral I/R,Frizzled-2 knockdown significantly lowered calcium concentration(P<0.001)and the expression levels of Wnt5a,Frizzled-2,and P-CaMK II in the IP area.In soy isoflavones-pretreated rats,calcium concentration,ROS and MDA levels,cell apoptosis rate,cerebral infarct volume,and expression levels of Wnt/Ca2+signaling pathway-related proteins were all significantly lower while SOD level was higher than those in rats in I/R model group.Conclusion Soy isoflavones can mitigate calcium overload in rats with cerebral I/R by inhibiting the Wnt/Ca2+signaling pathway.

Objective To explore the relationship between baseline urinary protein levels and the onset of chronic obstructive pulmonary disease (COPD). Methods A questionnaire survey, blood and urine sample collection, physical examination, and pulmonary function test were conducted among permanent residents over 40 years old in Pudong New Area, Shanghai. The subjects were divided into four groups based on the baseline urine albumin-to-creatinine ratio (ACR) quartiles (0~1.65 mg/g, 1.65~4.89 mg/g, 4.89~10.78 mg/g, and ≥10.78 mg/g). Cox regression analysis was used to explore the relationship between ACR levels and the incidence of COPD in middle-aged and elderly people. Results Among the 3 105 subjects, the median follow-up time was 3.212 years (P25~P75：3.102~3.473). 116 new cases of COPD were observed, with an incidence density of 10.423 per 1000 person-years. The incidence densities for COPD at four ACR levels were 7.922 per 1 000 person-years, 8.300 per 1 000 person-years , 11.419 per 1 000 person-years, and 13.843 per 1 000 person-years, respectively. Cox regression analysis revealed that as the ACR level increased, there was a rising trend in the incidence rate of COPD (χ²=4.396, P=0.036). After adjusting for gender, age, education level, occupational exposure to dust, history of childhood pneumonia, smoking, family history of COPD, central obesity, and hypertension, the risk of developing COPD was 2.499 times higher (95% CI: 1.460~4.276) for ACR levels ≥10.78 mg/g compared to the reference group with a baseline ACR level of 0~1.65 mg/g. Conclusion Elevated ACR levels in middle-aged and elderly population may increase the risk of COPD, and early monitoring of urine protein levels is beneficial for COPD prevention.

Objective:To establish the reference values and growth curves of skeletal muscle mass among children in the Nanjing area.Methods:A cross-sectional study was conducted with children who underwent physical examination at the Department of Child Health Care, Children′s Hospital of Nanjing Medical University from 2020 January to 2022 September. Their height, weight, body fat mass and skeletal muscle mass were measured. Body mass index, percentage of body fat mass, percentage of skeletal muscle mass, relative skeletal muscle mass index and the ratio of skeletal muscle to body fat were calculated. The associations between skeletal muscle mass indices and physical measurements index were analyzed through the Spearman correlation test. The Mann-Kendall test was used to assess the trend for skeletal muscle mass. Generalized additive models for location, scale and shape were used to construct percentile reference values and growth curves of male and female skeletal muscle mass indices at different ages.Results:A total of 32 690 children aged 4-14 years were enrolled in this study, including 19 912 boys (60.91%). Skeletal muscle mass, percentage of skeletal muscle mass, relative skeletal muscle mass index and the ratio of skeletal muscle to body fat of boys and girls was 11.10 (8.40, 14.90) and 10.30 (7.90, 13.20) kg, 40.36% (37.01%, 43.13%) and 39.38% (36.43%, 41.88%), 6.70 (6.07, 7.52) and 6.33 (5.79, 7.00), 2.39 (1.46, 3.47) and 2.14 (1.45, 3.00) kg/m 2, respectively. Skeletal muscle mass of both boys and girls was all positively associated with weight ( r=0.97, 0.96), body mass index ( r=0.68, 0.63) and percentage of body fat mass ( r=0.40, 0.43) (all P<0.01). The reference values and growth curves showed that the percentage of skeletal muscle mass P50 ranged from 37.75%-44.61% in boys and from 36.22%-40.55% in girls. The relative skeletal muscle mass index P50 ranged from 5.80-9.68 kg/m 2 in boys and from 5.57-7.98 kg/m 2 in girls. The ratio of skeletal muscle to body fat P50 ranged from 1.86-2.67 in boys and from 1.29-2.41 in girls. There was an increasing trend with age for both boys and girls in the growth of skeletal muscle mass ( Z=4.20, 3.75, both Ptrend<0.01), and increased slightly before 9 years of age and then increased rapidly until 14 years of age in both boys and girls. Conclusions:The skeletal muscle mass indices change with age and gender during childhood. Percentile reference values for pediatric skeletal muscle mass indices can be used to evaluate the muscular growth and development in children in the Nanjing area.

Objective:To investigate the predictive value of neck circumference on cardiometabolic risk in children.Methods:This was a cross-sectional study of natural sources. As the prediction cohort, clinical data were collected from 3 443 children aged 5-14 years who underwent physical examination in the Department of Child Healthcare, Children′s Hospital of Nanjing Medical University from July 2021 to September 2022. As the validation cohort for external validation, clinical data were collected from 604 children aged 5-14 years who underwent physical examination in the Department of Child Healthcare, Children′s Hospital of Nanjing Medical University from October 2022 to March 2023. Height, weight, neck circumference, waist circumference and body composition were measured in both groups, and body mass index, neck circumference to height ratio (NHtR), waist circumference to height ratio, body fat percentage and skeletal muscle percentage were calculated. Systolic blood pressure, diastolic blood pressure, fasting blood glucose, blood lipid and uric acid and other cardiovascular and metabolic risk indicators were collected in both groups. The prediction cohort was further stratified into clustered and non-clustered groups based on the clustering of cardiometabolic risk factors (CCRF). Various variables between these 2 groups were compared using the Mann-Whitney U test. Pearson correlation and binary Logistic regression were conducted to investigate the correlations between neck circumference and cardiovascular metabolic risk factors. The accuracy of NHtR in predicting the CCRF was evaluated using the area under the curve (AUC) of receiver operating characteristic (ROC). The cutoff value was determined using the Youden index. The validation cohort was then divided into groups above and below the cutoff value, and the detection rate of CCRF between the 2 groups was compared using the χ2 test for validation .Results:In the prediction cohort of 3 443 children (2 316 boys and 1 127 girls), 1 395 (40.5%) children were overweight or obese, and 1 157 (33.6%) children had CCRF. Pearson correlation analysis revealed all significant positive correlations (all P<0.01) between neck circumference and systolic blood pressure ( r=0.47, 0.39), diastolic blood pressure ( r=0.27, 0.21), uric acid ( r=0.36, 0.30), and triglycerides ( r=0.20, 0.20) after adjusting for age in both males and females. Among both males and females, neck circumference both showed significant negative correlation (both P<0.01) with high-density lipoprotein cholesterol ( r=-0.27, -0.28), and no correlation with fasting glucose levels ( r=0.03, -0.03, both P>0.05). After adjusting for gender, age, and body fat percentage, increased body mass index, neck circumference, or waist circumference increased the risks of hypertension ( OR=1.23, 1.39, 1.07, all P<0.001), hyperuricemia ( OR=1.16, 1.23, 1.05, all P<0.001), hypertriglyceridemia ( OR=1.08, 1.16, 1.02, all P<0.01), low high-density lipoprotein cholesterol ( OR=1.10, 1.27, 1.03, all P<0.01), and the CCRF ( OR=1.51, 1.73, 1.15, all P<0.01). The areas under the ROC curves of NHtR in predicting CCRF was 0.73, with sensitivity and specificity at 0.66 and 0.71, respectively. The corresponding optimal cut-off value was 0.21. Validation with 604 children confirmed that the detection of CCRF in the NHtR≥0.21 group was 3.29 times (60.5% (112/185) vs. 18.7% (79/422), χ2=107.82, P<0.01) higher compared to the NHtR <0.21 group. Conclusions:Neck circumference is associated with cardiovascular metabolic risks such as hypertension, hyperlipidemia, hyperglycemia, and hyperuricemia in children. When the NHtR is ≥0.21, there is an increased likelihood of CCRF.