Based on serum metabolomics and gut microbiota technology, this study explores the effects and mechanisms of the water extract of Ziziphi Spinosae Semen(SZRW) and the petroleum ether extract of Ziziphi Spinosae Semen(SZRO) in improving depressive-like behaviors induced by sleep deprivation. A modified multi-platform water environment method was employed to establish a rat model of sleep deprivation. Depressive-like behaviors in rats were assessed through the sucrose preference test and forced swim test. The expression of barrier proteins, such as Occludin, in the colon was determined by immunofluorescence. UPLC-Q-Orbitrap MS was utilized to analyze the serum metabolic profiles of sleep-deprived rats, screen for differential metabolites, and analyze metabolic pathways. The diversity of the gut microbiota was detected using 16S rRNA gene sequencing. Spearman correlation coefficient analysis was conducted to assess the correlation between differential metabolites and gut microbiota. The results indicated that SZRO significantly increased the sucrose preference index and decreased the immobility time in the forced swim test in rats. A total of 34 differential metabolites were identified through serum metabolomics. SZRW and SZRO shared five metabolic pathways, including phenylalanine metabolism. SZRW uniquely featured taurine and hypotaurine metabolism, while SZRO uniquely featured linoleic acid metabolism and tyrosine metabolism. Correlation analysis revealed that SZRW could upregulate the abundance of Bilophila, promoting the production of indole-3-propionic acid and subsequently upregulating the expression levels of intestinal tight junction proteins such as ZO-1, Occludin, and Claudin-1. SZRO could indirectly influence metabolic pathways such as arginine metabolism and linoleic acid metabolism by upregulating the abundance of gut microbiota such as Coprococcus and Eubacterium species. Both SZRW and SZRO can regulate endogenous metabolism, including amino acids, energy, and lipids, alter the gut microbiota microecology, and improve depressive-like behaviors. SZRO demonstrated superior effects in regulating metabolic pathways and gut microbiota structure compared to SZRW. The findings of this study provide a scientific basis for elucidating the pharmacodynamic material basis of Ziziphi Spinosae Semen.
Animals ; Rats ; Gastrointestinal Microbiome/drug effects* ; Male ; Metabolomics ; Drugs, Chinese Herbal/administration & dosage* ; Depression/blood* ; Rats, Sprague-Dawley ; Sleep Deprivation/complications* ; Ziziphus/chemistry* ; Antidepressive Agents/administration & dosage* ; Behavior, Animal/drug effects* ; Humans

The n-butanol fraction of Alpinia oxyphylla Fructus 70% ethanol extract was separated and purified using column chromatography with MCI Gel CHP-20, Sephadex LH-20, ODS, and silica gel, combined with semi preparative liquid phase and TLC separation methods. One new halogenated 4,5-seco-eudesmane sesquiterpenoid and two new eremophilane sesquiterpenoids were isolated and purified from the n-butanol fraction of Alpinia oxyphylla Fructus. The structures of the isolated compounds were identified using modern spectroscopic methods (1D, 2D NMR, UV, IR, MS, etc.), and the absolute configuration of the new compounds were determined using the methods of calculated ECD and induced ECD.

Atherosclerosis caused by disorders of lipid metabolism is the main pathological basis of atherosclerotic cardiovascular disease.Statins are the cornerstone of lipid-modulating therapy for this type of disease,but in practice there are still some patients with suboptimal lipid management.Proprotein convertase subtilisin/kexin type 9(PCSK9)inhibitors have been gradually applied as a new class of lipid-modulating drugs for the treatment in patients with this type of disease,and recent studies have shown that in addition to regulating lipid metabolism,PCSK9 inhibitors also have potential anti-inflammatory and anti-platelet activation effects.This article sorts out the multiple pharmacological mechanisms of action of PCSK9 inhibitors and the current status of clinical research of PCSK9 inhibitors.Besides,it discusses the factors that may affect the efficacy of PCSK9 inhibitors,in order to provide a reference for the safe and rational medication of PCSK9 inhibitors.

AIM To establish a two reference substances for determination of multiple components(TRSDMC)method for the simultaneous content determination of neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,luteolin-7-O-glucuronide,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,deoxyelephantopin,isodeoxyelephantopin,isoscabertopin and scabertopin in Elephantopus scabre L..METHODS The analysis was performed on a 35℃thermostatic Waters Symmetry C18,Phenomenex C18,Agilent ZORBAX Eclipse Plus C18 columns(4.6 mm×250 mm,5.0 μm),with the mobile phase comprising of acetonitrile and 0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 220,326 nm.Chlorogenic acid was used as an internal standard to calculate the relative correction factors of neochlorogenic acid,cryptochlorogenic acid,luteolin-7-O-glucuronide,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C,while isodeoxyelephantopin was used as an internal standard to calculate the relative correction factors of deoxyelephantopin,scabertopin and isoscabertopin,after which the content determination was made.Subsequently,cluster analysis,principal component analysis and orthogonal partial least squares-discriminant analysis were conducted.RESULTS Eleven constituents showed good linear relationships within their own ranges(r≥0.999 0),whose average recoveries were 95.3%-103.4%with the RSDs of 0.32%-3.45%.The result obtained by TRSDMC approximated those obtained by external standard method.Isochlorogenic acid A,isochlorogenic acid C,isochlorogenic acid B,chlorogenic acid,luteolin-7-O-glucuronide and cryptochlorogenic acid were taken as quality differential constituents.CONCLUSION This reliable and stable method can be used for the quality control of E.scabre.

Objective To evaluate healthcare-associated infection(HAI)status and influencing factors in coronary heart disease(CHD)patients after percutaneous coronary intervention(PCI)treatment,and construct a risk predic-tion model.Methods CHD patients who underwent PCI in a hospital from May 2019 to October 2023 were retro-spectively selected as the research subjects.Infection status of the CHD patients was analyzed.Patients were ran-domly divided into a modeling set and a testing set in a 7:3 ratio.Univariate and multivariate logistic regression ana-lyses were performed to analyze the data in the modeling set and determine the influencing factors for HAI in pa-tients.R software was used to construct and validate a nomogram model.Results A total of 858 CHD patients were included in the analysis,601 in the modeling set and 257 in the testing set.In the modeling set,41 cases were in the infected group and 560 cases in the non-infected group.The incidence of HAI in CHD patients after PCI treat-ment was 6.88％(59/858).Infection site were mainly upper respiratory tract and urinary tract.A total of 74 pathogens were isolated,including Gram-positive bacteria,Gram-negative bacteria,and fungi being 39,31,and 4 strains,respectively.Multivariate analysis showed that old age,combined diabetes,high grade of New York Heart Association(NYHA)classification,and invasive procedures were all risk factors for HAI in CHD patients after PCI treatment(all P＜0.05),while high mini-nutritional assessment short-form(MNA-SF)score was a protective fac-tor(P＜0.05).The area under the receiver operating characteristic(ROC)curve(AUC)of the nomogram predic-tion model constructed based on the above five indicators was 0.894(95％CI:0.815-0.931),with a sensitivity of 89.0％and a specificity of 82.5％.The testing set data validation showed an AUC value of 0.879(95％CI:0.801-0.923),with a sensitivity of 87.5％and a specificity of 81.3％,which were comparable to the modeling set and presented the stability of the model.The H-L goodness of fit test showed no statistical significance(P＞0.05),in-dicating that the model didn't exhibit overfitting.Calibration curve analysis showed that the model had good consis-tency.Decision curve analysis confirmed that the model had practical value in clinical practice.Conclusion The no-mogram model has a good predictive ability for HAI in CHD patients after PCI treatment,and can provide a simple and effective evaluation tool for medical staff to identify HAI high-risk individuals.

Objective:To explore the clinical value of high mobility group box-1（HMGB1）and neutrophil elastase（NE）in early detection of Kawasaki disease（KD）and prediction of coronary artery lesions（CAL）.Method:In this case-control study，80 children with Kawasaki disease who were hospitalized in the Department of Cardiovascular Medicine，Xuzhou Children's Hospital，Xuzhou Medical University from September 2022 to October 2023 were selected as the KD group，and they were further divided into the CAL subgroup and the non-CAL（NCAL）subgroup according to the color echocardiography results of coronary artery.Additionally，40 children with fever diagnosed as respiratory infectious diseases and 40 healthy children for general health check-ups during the same period were enrolled as the fever control group and the healthy control group，respectively.The one-way ANOVA，least significant difference（LSD）method and Bonferroni correction were used to compare the serum HMGB1 and NE levels in KD group，fever control group and healthy control group as well as the changes of serum HMGB1 and NE levels in acute，sub-acute and convalescence stages of KD group.The independent sample t-test was applied to compare the differences between CAL subgroup and NCAL subgroup at acute，sub-acute and convalescence stages.The receiver operating characteristic curve was drawn to analyze the value of serum HMGB1 and NE for early detection of KD and prediction of KD with CAL. Results:In the acute stage，the serum levels of HMGB1 and NE of children with KD were significantly higher than those of the two control groups，with statistically significant differences（ F=60.962，45.805，all P<0.05）.The serum levels of HMGB1 and NE of the KD group were higher in the acute stage than those in the sub-acute stage and the convalescence stage，with statistically significant differences（ F=14.369，26.641，all P<0.05）.The serum levels of HMGB1 and NE of the CAL subgroup were higher than those of the NCAL subgroup in the acute，sub-acute and convalescence stage，with statistically significant differences（ t=-3.086，-3.055，-3.321，-5.212，-2.647，-2.345，all P<0.05）.The optimal critical values of HMGB1 and NE for early detection of KD were 2.04 μg/L and 241.00 μg/L，respectively.The area under curves（AUC）of serum HMGB1 and NE and integrated examination for early detection of KD were 0.89，0.85，and 0.90，respectively；the corresponding sensitivities were 87.50%，71.30%，and 75.00%，respectively，and the specificities were 81.20%，90.00%，and 92.50%，respectively.In the acute stage of KD group，the optimal critical values of serum HMGB1 and NE for prediction of CAL were 2.60 μg/L and 261.00 μg/L，respectively.The AUC of serum HMGB1 and NE and integrated examination for prediction of CAL were 0.70，0.80，and 0.81，respectively；the corresponding sensitivities were 63.60%，86.40%，and 72.70%，respectively，and the specificities were 70.70%，67.20%，and 77.60%，respectively. Conclusions:The increased levels of serum HMGB1 and NE in KD acute stage have certain reference value for the diagnosis of KD；HMGB1 and NE have a certain role in predicting KD with CAL，and they are of greater value when they are combined for detection.

METHODS: Rat BMSCs were isolated, cultured and identified. The time-dependent expressions of TSP-2 and Sox9 in BMSCs under a dynamic mechanical pressure of 0–120 kPa at 0.1 Hz for 1 h were tested by qPCR and Western blotting. The role of TSP-2 in chondrogenic differentiation of BMSCs under mechanical pressure was validated by using small interfering RNA. The impact of TSP-2 and mechanical pressure on chondrogenesis were detected and the downstream signaling molecules were explored using Western blotting. RESULTS: Mechanical pressure stimulation of 0–120 kPa for 1 h significantly upregulated the expression of TSP-2 in BMSCs. The expression of the chondrogenesis markers Sox9, Aggrecan, and Col-II were all upregulated under dynamic mechanical pressure or TSP-2 stimulation. Additional exogenous TSP-2 may potentiate the chondrogenic effect of mechanical stimulation. After knock down TSP-2, the upregulation of Sox9, Aggrecan and Col-II under mechanical pressure was inhibited. The NF-jB signaling pathway responded to both dynamic pressure and TSP-2 stimulation, and the cartilage-promoting effect was blocked by an NF-jB signaling inhibitor. CONCLUSION TSP-2 plays an essential role in the chondrogenic differentiation of BMSCs under mechanical pressure. NF-jB signaling is involved in the mechano-chemical coupling of TSP-2 and mechanical pressure for the chondrogenic differentiation of BMSCs.

Objective To investigate the influencing factors and establish a model predicting the performance of needle visualization in fine-needle aspiration (FNA) of thyroid nodules. Methods This study prospectively included 175 patients who underwent FNA of thyroid nodules in the Department of Ultrasound in China-Japan Friendship Hospital and compared the display of the needle tips in the examination of 199 thyroid nodules before and after the application of needle visualization.We recorded the location,the positional relationship with thyroid capsule,ultrasonic characteristics,and the distribution of the soft tissue strip structure at the puncture site of the nodules with unclear needle tips display before using needle visualization.Furthermore,according to the thyroid imaging reporting and data system proposed by the American College of Radiology,we graded the risk of the nodules.Lasso-Logistic regression was employed to screen out the factors influencing the performance of needle visualization and establish a nomogram for prediction. Results The needle tips were not clearly displayed in the examination of 135 (67.8%) and 53 (26.6%) nodules before and after the application of needle visualization,respectively,which showed a significant difference (P<0.001).Based on the positional relationship between the nodule and capsule,anteroposterior/transverse diameter (A/T) ratio,blood supply,and the distribution of subcutaneous strip structure at the puncture site,a nomogram was established to predict the probability of unclear display of the needle tips after application of needle visualization.The C-index of the prediction model was 0.75 (95%CI=0.67-0.84) and the area under the receiver operating characteristic curve was 0.72.The calibration curve confirmed the appreciable reliability of the prediction model,with the C-index of 0.70 in internal validation. Conclusions Needle visualization can improve the display of the needle tip in ultrasound-guided FNA of thyroid nodules.The nomogram established based on ultrasound features such as the positional relationship between the nodule and capsule,A/T ratio,blood supply,and the distribution of subcutaneous strip structure at the puncture site can predict whether needle visualization is suitable for the examination of nodules.
Humans ; Thyroid Nodule/diagnostic imaging* ; Biopsy, Fine-Needle/methods* ; Reproducibility of Results ; Ultrasonography ; Retrospective Studies ; Thyroid Neoplasms

Hepatic lipid deposition is one of the basic manifestations of obesity, and nowadays pharmacological treatment is the most important tool. Punicalagin(PU), a polyphenol derived from pomegranate peel, is a potential anti-obesity substance. In this study, 60 C57BL/6J mice were randomly divided into a normal group and a model group. After establishing a model of simple obesity with a high-fat diet for 12 weeks, the successfully established rat models of obesity were then regrouped into a model group, an orlistat group, a PU low-dose group, a PU medium-dose group, and a PU high-dose group. The normal group was kept on routine diet and other groups continued to feed the high-fat diet. The body weight and food intake were measured and recorded weekly. After 8 weeks, the levels of the four lipids in the serum of each group of mice were determined by an automatic biochemical instrument. Oral glucose tole-rance and intraperitoneal insulin sensitivity were tested. Hemoxylin-eosin(HE) staining was applied to observe the hepatic and adipose tissues. The mRNA expression levels of peroxisome proliferators-activated receptor γ(PPARγ) and C/EBPα were determined by real-time quantitative polymerase chain reaction(Q-PCR), and the mRNA and protein expression levels of adenosine 5'-monophosphate-activated protein kinase(AMPK), anterior cingulate cortex(ACC), and carnitine palmitoyltransferase 1A(CPT1A) were determined by Western blot. Finally, the body mass, Lee's index, serum total glyceride(TG), serum total cholesterol(TC), and low-density lipoprotein cholesterol(LDL-C) levels were significantly higher and high-density lipoprotein cholesterol(HDL-C) levels were significantly lower in the model group as compared with the normal group. The fat deposition in the liver was significantly increased. The mRNA expression levels of hepatic PPARγ and C/EBPα and the protein expression level of ACC were increased, while the mRNA and protein expression levels of CPT-1α(CPT1A) and AMPK were decreased. After PU treatment, the above indexes of obese mice were reversed. In conclusion, PU can decrease the body weight of obese mice and control their food intake. It also plays a role in the regulation of lipid metabolism and glycometabolism metabolism, which can significantly improve hepatic fat deposition. Mechanistically, PU may regulate liver lipid deposition in obese mice by down-regulating lipid synthesis and up-regulating lipolysis through activation of the AMPK/ACC pathway.
Rats ; Mice ; Animals ; Mice, Obese ; AMP-Activated Protein Kinases/metabolism* ; PPAR gamma/metabolism* ; Mice, Inbred C57BL ; Liver/metabolism* ; Obesity/genetics* ; Body Weight ; Lipid Metabolism ; Diet, High-Fat/adverse effects* ; Lipids ; Cholesterol

OBJECTIVES: To study the clinical features and fibroblast growth factor receptor 3 (FGFR3) gene mutations of children with achondroplasia (ACH) through an analysis of 17 cases. METHODS: A retrospective analysis was performed on the clinical data and FGFR3 gene detection results of 17 children with ACH who were diagnosed from January 2009 to October 2021. RESULTS: Of the 17 children with ACH, common clinical manifestations included disproportionate short stature (100%, 17/17), macrocephaly (100%, 17/17), trident hand (82%, 14/17), and genu varum (88%, 15/17). The common imaging findings were rhizomelic shortening of the long bones (100%, 17/17) and narrowing of the lumbar intervertebral space (88%, 15/17). Major complications included skeletal dysplasia (100%, 17/17), middle ear dysfunction (82%, 14/17), motor/language developmental delay (88%, 15/17), chronic pain (59%, 10/17), sleep apnea (53%, 9/17), obesity (41%, 7/17), foramen magnum stenosis (35%, 6/17), and hydrocephalus (24%, 4/17). All 17 children (100%) had FGFR3 mutations, among whom 13 had c.1138G>A hotspot mutations of the FGFR3 gene, 2 had c.1138G>C mutations of the FGFR3 gene, and 2 had unreported mutations, with c.1252C>T mutations of the FGFR3 gene in one child and c.445+2_445+5delTAGG mutations of the FGFR3 gene in the other child. CONCLUSIONS This study identifies the unreported mutation sites of the FGFR3 gene, which extends the gene mutation spectrum of ACH. ACH is a progressive disease requiring lifelong management through multidisciplinary collaboration.
Achondroplasia/genetics* ; Child ; Humans ; Mutation ; Osteochondrodysplasias/genetics* ; Receptor, Fibroblast Growth Factor, Type 3/genetics* ; Retrospective Studies