This study aims to investigate the antipyretic effects and mechanisms of ethanol extracts from Arisaematis Rhizoma fermented with bile from different sources on a rat model of fever induced by a dry-yeast suspension. The rat model of fever was established by subcutaneous injection of 20% dry-yeast suspension into the rat back. The levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6) in the serum, as well as prostaglandin E_2(PGE_2) and cyclic adenosine monophosphate(cAMP) in the hypothalamus, were determined by ELISA. Metabolomics analysis was then performed on serum and hypothalamus samples based on UPLC-Q-TOF MS to explore the potential biomarkers and metabolic pathways. The results showed that the body temperatures of rats significantly rose 4 h after modeling. After oral administration of high-dose ethanol extracts of Arisaematis Rhizoma fermented with bovine bile(NCH) and porcine bile(ZCH), the body temperatures of rats declined(P<0.05), and the NCH group showed better antipyretic effect than the ZCH group. Additionally, compared with the model group, the NCH and ZCH groups showed lowered levels of IL-1β, IL-6, TNF-α, PGE_2, and cAMP(P<0.01). The results of serum and hypothalamus metabolomics analysis indicated that both NCH and ZCH exerted antipyretic effects by regulating phenylalanine metabolism, sphingolipid metabolism, arachidonic acid metabolism, and steroid hormone biosynthesis. Collectively, both NCH and ZCH can play an obvious antipyretic role in the rat model of dry yeast-induced fever, and the underlying mechanism might be closely associated with inhibiting inflammation and regulating metabolic disorders. Moreover, NCH demonstrates better antipyretic effect.
Animals ; Rats ; Male ; Fermentation ; Rats, Sprague-Dawley ; Rhizome/metabolism* ; Drugs, Chinese Herbal/chemistry* ; Bile/chemistry* ; Antipyretics/chemistry* ; Fever/metabolism* ; Cattle ; Swine ; Tumor Necrosis Factor-alpha/metabolism* ; Ethanol/chemistry* ; Interleukin-6/blood* ; Interleukin-1beta/blood*

Based on the high-fat diet-induced hyperlipidemia rat model, this study aimed to evaluate the lipid-lowering effect of Arisaema Cum Bile and explore its mechanisms, providing experimental evidence for its clinical application. Biochemical analysis was used to detect serum levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), triglycerides(TG), and total cholesterol(TC) to assess the lipid-lowering activity of Arisaema Cum Bile. Additionally, 16S rDNA sequencing and metabolomics techniques were employed to jointly elucidate the lipid-lowering mechanisms of Arisaema Cum Bile. The experimental results showed that high-dose Arisaema Cum Bile(PBA-H) significantly reduced serum ALT, AST, LDL-C, TG, and TC levels(P<0.01), and significantly increased HDL-C levels(P<0.01). The effect was similar to that of fenofibrate, with no significant difference. Furthermore, Arisaema Cum Bile significantly alleviated hepatocyte ballooning and mitigated fatty degeneration in liver tissues. As indicated by 16S rDNA sequencing results, PBA-H significantly enhanced both alpha and beta diversity of the gut microbiota in the model rats, notably increasing the relative abundance of Akkermansia and Subdoligranulum species(P<0.01). Liver metabolomics analysis revealed that PBA-H primarily regulated pathways involved in arachidonic acid metabolism, vitamin B_6 metabolism, and steroid biosynthesis. In summary, Arisaema Cum Bile significantly improved abnormal blood lipid levels and liver pathology induced by a high-fat diet, regulated hepatic metabolic disorders, and improved the abundance and structural composition of gut microbiota, thereby exerting its lipid-lowering effect. The findings of this study provide experimental evidence for the clinical application of Arisaema Cum Bile and the treatment of hyperlipidemia.
Animals ; Gastrointestinal Microbiome/drug effects* ; Rats ; Male ; Metabolomics ; Hyperlipidemias/microbiology* ; Drugs, Chinese Herbal/administration & dosage* ; Rats, Sprague-Dawley ; Hypolipidemic Agents/pharmacology* ; Liver/metabolism* ; Humans ; Alanine Transaminase/metabolism* ; Triglycerides/metabolism* ; Aspartate Aminotransferases/metabolism*

Anxiety disorder is a major symptom of autism spectrum disorder (ASD) with a comorbidity rate of ~40%. However, the neural mechanisms of the emergence of anxiety in ASD remain unclear. In our study, we found that hyperactivity of basolateral amygdala (BLA) pyramidal neurons (PNs) in Shank3 InsG3680 knock-in (InsG3680^+/+) mice is involved in the development of anxiety. Electrophysiological results also showed increased excitatory input and decreased inhibitory input in BLA PNs. Chemogenetic inhibition of the excitability of PNs in the BLA rescued the anxiety phenotype of InsG3680^+/+ mice. Further study found that the diminished control of the BLA by medial prefrontal cortex (mPFC) and optogenetic activation of the mPFC-BLA pathway also had a rescue effect, which increased the feedforward inhibition of the BLA. Taken together, our results suggest that hyperactivity of the BLA and alteration of the mPFC-BLA circuitry are involved in anxiety in InsG3680^+/+ mice.
Animals ; Prefrontal Cortex/metabolism* ; Basolateral Nuclear Complex/metabolism* ; Mice ; Anxiety/metabolism* ; Nerve Tissue Proteins/genetics* ; Male ; Gene Knock-In Techniques ; Pyramidal Cells/physiology* ; Mice, Transgenic ; Neural Pathways/physiopathology* ; Mice, Inbred C57BL ; Microfilament Proteins

Objective·[18F]F-FMISO and[18F]F-FLT are specific PET imaging agents for detecting the hypoxia microenvironment and cell proliferation,respectively.This study aims to visualize and monitor the impact of drug resistance in pancreatic cancer on the hypoxia microenvironment and cell proliferation through[18F]F-FMISO and[18F]F-FLT PET/CT dual-nuclide imaging,with the goal of providing a theoretical basis for clinical application.Methods·The CCK-8 assay was conducted to assess drug resistance in the PANC-1/R(PR)pancreatic cancer cell line compared to the parental PANC-1(P)cell line.Subcutaneous xenograft models of pancreatic cancer were established by injecting male BALB/c nude mice with pancreatic cancer cells into the left axillary subcutaneous region.Subgroups were treated with gemcitabine(GEM)chemotherapy starting on day 18(18D-G group)or day 12(12D-G group)after inoculation of tumor cells.[18F]F-FMISO and[18F]F-FLT PET/CT imaging were performed before and after treatment to obtain semi-quantitative parameters(maximum standardized uptake value,SUVmax).ΔSUVmax was calculated by using the following equation:ΔSUVmax=(SUVmax of second imaging-SUVmax of first imaging)/SUVmax of first imaging.Receiver operating characteristic(ROC)curves were used to determine the optimal threshold for the semi-quantitative parameters to assess pancreatic cancer drug resistance.Results·The CCK-8 assay confirmed that the PR cells exhibited high resistance to GEM,with a resistance index of 4.24(n=5).In vivo experiments showed that GEM chemotherapy significantly inhibited tumor growth and prolonged survival in the parental pancreatic cancer group(12D-G group,P=0.025),whereas GEM chemotherapy accelerated tumor growth and shortened survival(18D-G and 12D-G,P=0.025)in the drug-resistant pancreatic cancer group.In addition,in the non-chemotherapy group,ΔSUVmax-FLT might be negatively correlated with survival time,while in the chemotherapy group,both ΔSUVmax-FMISO and ΔSUVmax-FLT were negatively correlated with survival time(P=0.050,P=0.006).In the 18D-G and chemotherapy group,the second imaging showed significantly lower ΔSUVmax-FMISO and ΔSUVmax-FLT in P tumors compared to PR tumors(P=0.045,P=0.050).In the 12D-G and chemotherapy group,the second imaging showed slightly lower ΔSUVmax-FLT in P tumors compared to PR tumors(P=0.051).ROC analysis identified the optimal threshold for assessing pancreatic cancer drug resistance:when ΔSUVmax-FLT=0.45 in the non-chemotherapy group,the sensitivity and specificity were 100.00%and 50.00%,respectively;when ΔSUVmax-FMISO=0.37 and ΔSUVmax-FLT=0.36 in the chemotherapy group,the sensitivity and specificity were 100.00%and 83.33%,respectively.Conclusion·[18F]F-FMISO and[18F]F-FLT PET/CT dual-nuclide imaging can be used to assess drug resistance in pancreatic cancer.The comparison of[18F]F-FMISO and[18F]F-FLT PET differences before and after chemotherapy provides the most accurate prediction of drug resistance and survival time.

Objective To explore the effect and predictive value of ultrasound-guided needling assisted motor evoked potentials(MEP)and electromyography(EMG)monitoring on neurological recovery in spinal surgery.Methods A retrospective analysis was conducted on the clinical data of 80 patients who underwent spinal surgery at Jiangsu Province Hospital of Chinese Medicine from January 2020 to December 2024.A total of 41 patients in the observation group received ultrasound-guided needling assisted MEP and EMG monitoring,and 39 patients in the control group received conventional method for MEP and EMG monitoring.The operative time,intraoperative blood loss,and the proportions of intraoperative MEP and EMG warnings were compared between the two groups,and the sensitivity and specificity of intraoperative MEP monitoring were compared between the two groups.The receiver operating characteristic(ROC)curve was plotted,and the area under the curve(AUC)was calculated to analyze the efficiency of MEP warning in predicting the dysfunction of postoperative spinal cord.Results There were no significant differences in the operative time,intraoperative blood loss,or the proportions of intraoperative MEP and EMG warnings(P>0.05).The sensitivity,specificity and AUC of intraoperative MEP monitoring in the observation group were significantly higher than those in the control group,with statistically significant differences(P<0.05).The sensitivity,specificity,and AUC of postoperative MEP warning in predicting the dysfunction of spinal cord in the observation group were higher than those in the control group,with statistically significant differences(P<0.05).Conclusion Ultrasound-guided needling assisted MEP and EMG monitoring can effectively enhance the intraoperative neural monitoring accuracy,and postoperative MEP warning demonstrates superior predictive value for postoperative neurological dysfunction.

Objective To explore the effect and predictive value of ultrasound-guided needling assisted motor evoked potentials(MEP)and electromyography(EMG)monitoring on neurological recovery in spinal surgery.Methods A retrospective analysis was conducted on the clinical data of 80 patients who underwent spinal surgery at Jiangsu Province Hospital of Chinese Medicine from January 2020 to December 2024.A total of 41 patients in the observation group received ultrasound-guided needling assisted MEP and EMG monitoring,and 39 patients in the control group received conventional method for MEP and EMG monitoring.The operative time,intraoperative blood loss,and the proportions of intraoperative MEP and EMG warnings were compared between the two groups,and the sensitivity and specificity of intraoperative MEP monitoring were compared between the two groups.The receiver operating characteristic(ROC)curve was plotted,and the area under the curve(AUC)was calculated to analyze the efficiency of MEP warning in predicting the dysfunction of postoperative spinal cord.Results There were no significant differences in the operative time,intraoperative blood loss,or the proportions of intraoperative MEP and EMG warnings(P>0.05).The sensitivity,specificity and AUC of intraoperative MEP monitoring in the observation group were significantly higher than those in the control group,with statistically significant differences(P<0.05).The sensitivity,specificity,and AUC of postoperative MEP warning in predicting the dysfunction of spinal cord in the observation group were higher than those in the control group,with statistically significant differences(P<0.05).Conclusion Ultrasound-guided needling assisted MEP and EMG monitoring can effectively enhance the intraoperative neural monitoring accuracy,and postoperative MEP warning demonstrates superior predictive value for postoperative neurological dysfunction.

Objective·[18F]F-FMISO and[18F]F-FLT are specific PET imaging agents for detecting the hypoxia microenvironment and cell proliferation,respectively.This study aims to visualize and monitor the impact of drug resistance in pancreatic cancer on the hypoxia microenvironment and cell proliferation through[18F]F-FMISO and[18F]F-FLT PET/CT dual-nuclide imaging,with the goal of providing a theoretical basis for clinical application.Methods·The CCK-8 assay was conducted to assess drug resistance in the PANC-1/R(PR)pancreatic cancer cell line compared to the parental PANC-1(P)cell line.Subcutaneous xenograft models of pancreatic cancer were established by injecting male BALB/c nude mice with pancreatic cancer cells into the left axillary subcutaneous region.Subgroups were treated with gemcitabine(GEM)chemotherapy starting on day 18(18D-G group)or day 12(12D-G group)after inoculation of tumor cells.[18F]F-FMISO and[18F]F-FLT PET/CT imaging were performed before and after treatment to obtain semi-quantitative parameters(maximum standardized uptake value,SUVmax).ΔSUVmax was calculated by using the following equation:ΔSUVmax=(SUVmax of second imaging-SUVmax of first imaging)/SUVmax of first imaging.Receiver operating characteristic(ROC)curves were used to determine the optimal threshold for the semi-quantitative parameters to assess pancreatic cancer drug resistance.Results·The CCK-8 assay confirmed that the PR cells exhibited high resistance to GEM,with a resistance index of 4.24(n=5).In vivo experiments showed that GEM chemotherapy significantly inhibited tumor growth and prolonged survival in the parental pancreatic cancer group(12D-G group,P=0.025),whereas GEM chemotherapy accelerated tumor growth and shortened survival(18D-G and 12D-G,P=0.025)in the drug-resistant pancreatic cancer group.In addition,in the non-chemotherapy group,ΔSUVmax-FLT might be negatively correlated with survival time,while in the chemotherapy group,both ΔSUVmax-FMISO and ΔSUVmax-FLT were negatively correlated with survival time(P=0.050,P=0.006).In the 18D-G and chemotherapy group,the second imaging showed significantly lower ΔSUVmax-FMISO and ΔSUVmax-FLT in P tumors compared to PR tumors(P=0.045,P=0.050).In the 12D-G and chemotherapy group,the second imaging showed slightly lower ΔSUVmax-FLT in P tumors compared to PR tumors(P=0.051).ROC analysis identified the optimal threshold for assessing pancreatic cancer drug resistance:when ΔSUVmax-FLT=0.45 in the non-chemotherapy group,the sensitivity and specificity were 100.00%and 50.00%,respectively;when ΔSUVmax-FMISO=0.37 and ΔSUVmax-FLT=0.36 in the chemotherapy group,the sensitivity and specificity were 100.00%and 83.33%,respectively.Conclusion·[18F]F-FMISO and[18F]F-FLT PET/CT dual-nuclide imaging can be used to assess drug resistance in pancreatic cancer.The comparison of[18F]F-FMISO and[18F]F-FLT PET differences before and after chemotherapy provides the most accurate prediction of drug resistance and survival time.

Objective To explore the molecular epidemiological characteristics of carbapenem-resistant Klebsiella pneumoniae(CRKP),and reveal its mechanism of resistance to ceftazidime/avibactam(CZA).Methods CZA-re-sistant CRKP strains initially isolated from the Affiliated Hospital of Xuzhou Medical University from January 2021 to September 2023 were collected.The carriage of 5 carbapenemase genes(blaKPC,blaNDM,blaOXA,blaVIM,blaIMp)were detected with gene amplification method and colloidal gold method.The relative copy number and expression level of Klebsiella pneumoniae(KP)carbapenemase-producing KP(KPC-KP)was detected with real-time quantita-tive polymerase chain reaction(RT-qPCR),mutation sites of KPC mutation strains were analyzed with whole-ge-nome sequencing,and epidemic characteristics of CRKP and resistance mechanism to CZA were analyzed.Results A total of 73 CZA-resistant CRKP strains were isolated,with 37(50.68％)being KPC and NDM co-producing strains,33(45.21％)NDM-producing alone(23 strains producing NDM-5 and 10 strains producing NDM-1),and 3 KPC-producing alone.KP-2842 strain was identified as ST11-type KPC-33 variant,KP-2127 and KP-2189 strains produced KPC-2.Compared with KP ATCC BAA-1705,the copy number of blaKPC in these strains up-regulated by 1.04-3.86 fold,and the expression increased by 6.66-12.93 fold,respectively.Colloidal gold and PCR methods demonstrated good consistency and the ability to detect the enzyme co-producing and KPC-33 variant.Conclusion In this hospital,the resistance of CRKP to CZA is primarily mediated by the metalloenzyme NDM,with co-produc-tion of NDM and KPC being a characteristic of CRKP.High copy number and expression level of blaKPC-2 also con-tribute to CZA resistance.This study identified the KPC-33 variant for the first time in ST11-type CRKP in Jiangsu Province.

Lipopolysaccharides (LPS) are major outer membrane components of Gram-negative bacteria. Unlike most Gram-negative bacteria, Acinetobacter baumannii can still survive and acquire polymyxin resistance after complete loss of LPS. Previous studies of LPS-deficient Acinetobacter baumannii mostly focused on LPS-deficient strains induced by polymyxin, whose background was complex and unstable. To investigate LPS loss-mediated polymyxin resistant-Acinetobacter baumannii, this study constructed a stable and clear background LPS-deficient strain by knocking out lpxC gene in Acinetobacter baumannii ATCC 19606 with CIRSPR/Cas9, and then studied the phenotypic changes of the lpxC-deficient strain including morphology, growth rate, antibiotic susceptibility, virulence, membrane permeability, and membrane potential. Animal experiments were approved by the Animal Care and Welfare Committee Institute of Medicinal Biotechnology, CAMS and PUMC (approval number: IMB-20240119D₉). The results indicated that the lpxC gene of Acinetobacter baumannii ATCC 19606 was successfully knocked out. After losing the lpxC, the strain underwent the morphological change from rod-shaped to spherical. Furthermore, it leads to reduced growth rate, enhanced membrane permeability, decreased membrane potential, lower virulence, and increased antibiotic susceptibility to β-lactams, quinolones, aminoglycosides, macrolides, glycopeptides. The lpxC deletion results in significant changes in membrane homeostasis and adaptability of Acinetobacter baumannii. Understanding the phenotypic changes of colistin-resistant Acinetobacter baumannii mediated by LPS loss is useful for exploring the resistance mechanism of Acinetobacter baumannii and developing new therapeutic strategies.

Ankylosing spondylitis (AS) combined with spinal fractures with thoracic and lumbar fracture as the most common type shows characteristics of unstable fracture, high incidence of nerve injury, high mortality and high disability rate. The diagnosis may be missed because it is mostly caused by low-energy injury, when spinal rigidity and osteoporosis have a great impact on the accuracy of imaging examination. At the same time, the treatment choices are controversial, with no relevant specifications. Non-operative treatments can easily lead to bone nonunion, pseudoarthrosis and delayed nerve injury, while surgeries may be failed due to internal fixation failure. At present, there are no evidence-based guidelines for the diagnosis and treatment of AS combined with thoracic and lumbar fracture. In this context, the Spinal Trauma Academic Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate the Clinical guideline for the diagnosis and treatment of adult ankylosing spondylitis combined with thoracolumbar fracture ( version 2023) by following the principles of evidence-based medicine and systematically review related literatures. Ten recommendations on the diagnosis, imaging evaluation, classification and treatment of AS combined with thoracic and lumbar fracture were put forward, aiming to standardize the clinical diagnosis and treatment of such disorder.