ObjectiveTo investigate the efficacy and mechanism of Polygoni Cuspidati Rhizoma et Radix （Huzhang） in treating respiratory syncytial virus（RSV） infection by regulating pulmonary surfactant lipid homeostasis through lipidomics. MethodsSixty BALB/c mice were randomly divided into the blank group， model group， positive group（ribavirin group， 46 mg·kg^-1）， and low- and high-dose Huzhang groups（0.75， 2.25 g·kg^-1）， with 12 mice in each group. Except for the blank group， all other groups were infected with RSV via intranasal instillation. The drug intervention groups were given corresponding doses of drug by gavage for 3 consecutive days， while normal saline was used in the blank and model groups. Hematoxylin-eosin（HE） staining was used to observe pathological changes in mouse lung tissue. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was employed to detect viral loads［RSV-nucleoprotein（N） and RSV-glycoprotein（G） mRNA］ and inflammatory factor levels［interleukin（IL）-1β and tumor necrosis factor（TNF）-α mRNA］ in the lung tissue. Mouse bronchoalveolar lavage fluid was collected to detect the levels of pulmonary surfactant lipids through ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap-MS）， followed by principal component analysis and differential lipid identification. ResultsCompared with the blank group， the model group exhibited extensive inflammatory cell infiltration， congestion， and tissue damage in the lungs， and the pathological score and lung index of lung tissue significantly increased（P<0.01）， along with significantly elevated mRNA expressions of RSV-N， RSV-G， IL-1β， and TNF-α（P<0.01）. Compared with the model group， different doses of Huzhang and ribavirin significantly reduced the pathological scores of the lung tissue and lung index（P<0.01）. In addition， the mRNA levels of RSV-N， RSV-G and TNF-α in the lungs significantly decreased in the Huzhang high dose group（P<0.01）. Lipidomics analysis identified multiple significantly changed differential metabolites. Compared with the blank group， the model group showed obvious abnormal lipid metabolism， which was manifested by the elevated levels of prostaglandin（PG）， ceramide（Cer）， phosphatidylcholine（PC）， phosphatidylethanolamine（PE）， phosphatidylinositol（PI）， sphingomyelin（SM）， and the decreased levels of diglycerides（DG） and acylethanolamine（NAE）. After the intervention of low dose of Huzhang， the above lipid metabolites showed a significant reversal trend， while the intervention of high dose of Huzhang could regulate levels of PI lipids， PG lipids and PC lipids. ConclusionHuzhang can significantly reduce the viral load of lung tissue and improve lung inflammation in RSV-infected mice. The underlying mechanism may be related to the maintenance of homeostasis in pulmonary surfactant lipids such as PI and PG.

ObjectiveTo investigate the efficacy and mechanism of Polygoni Cuspidati Rhizoma et Radix （Huzhang） in treating respiratory syncytial virus（RSV） infection by regulating pulmonary surfactant lipid homeostasis through lipidomics. MethodsSixty BALB/c mice were randomly divided into the blank group， model group， positive group（ribavirin group， 46 mg·kg^-1）， and low- and high-dose Huzhang groups（0.75， 2.25 g·kg^-1）， with 12 mice in each group. Except for the blank group， all other groups were infected with RSV via intranasal instillation. The drug intervention groups were given corresponding doses of drug by gavage for 3 consecutive days， while normal saline was used in the blank and model groups. Hematoxylin-eosin（HE） staining was used to observe pathological changes in mouse lung tissue. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was employed to detect viral loads［RSV-nucleoprotein（N） and RSV-glycoprotein（G） mRNA］ and inflammatory factor levels［interleukin（IL）-1β and tumor necrosis factor（TNF）-α mRNA］ in the lung tissue. Mouse bronchoalveolar lavage fluid was collected to detect the levels of pulmonary surfactant lipids through ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap-MS）， followed by principal component analysis and differential lipid identification. ResultsCompared with the blank group， the model group exhibited extensive inflammatory cell infiltration， congestion， and tissue damage in the lungs， and the pathological score and lung index of lung tissue significantly increased（P<0.01）， along with significantly elevated mRNA expressions of RSV-N， RSV-G， IL-1β， and TNF-α（P<0.01）. Compared with the model group， different doses of Huzhang and ribavirin significantly reduced the pathological scores of the lung tissue and lung index（P<0.01）. In addition， the mRNA levels of RSV-N， RSV-G and TNF-α in the lungs significantly decreased in the Huzhang high dose group（P<0.01）. Lipidomics analysis identified multiple significantly changed differential metabolites. Compared with the blank group， the model group showed obvious abnormal lipid metabolism， which was manifested by the elevated levels of prostaglandin（PG）， ceramide（Cer）， phosphatidylcholine（PC）， phosphatidylethanolamine（PE）， phosphatidylinositol（PI）， sphingomyelin（SM）， and the decreased levels of diglycerides（DG） and acylethanolamine（NAE）. After the intervention of low dose of Huzhang， the above lipid metabolites showed a significant reversal trend， while the intervention of high dose of Huzhang could regulate levels of PI lipids， PG lipids and PC lipids. ConclusionHuzhang can significantly reduce the viral load of lung tissue and improve lung inflammation in RSV-infected mice. The underlying mechanism may be related to the maintenance of homeostasis in pulmonary surfactant lipids such as PI and PG.

Objective:To establish a method for detecting serum non-ceruloplasmin-bound copper (NCC) by immunoprecipitation-inductively coupled plasma mass spectrometry (IP-ICP-MS) and evaluate its clinical application.Methods:Methodological evaluation research. Immunoprecipitation was first used to separate serum ceruloplasmin, followed by detection of serum NCC levels using ICP-MS. Two levels of quality control serum were reconstituted to determine the limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision of the self-developed method. The serum samples of 131 healthy individuals (healthy group) and 69 first-time diagnosed Wilson′s disease (WD) patients(WD group) from November 2023 to June 2024 in the Affiliated Hospital of Institute of Neurology, Anhui University of Traditional Chinese Medicine were collected. Using self-developed method detected the serum NCC levels, established the healthy reference intervals, and NCC levels between the two groups were compared using the Mann-Whitney U test. Results:The calibration curve exhibited excellent lineary across the concentration range of 1.562 to 300.000 μg/L, as demonstrated by coefficient of determination ( R2) of 0.999. The self-developed NCC method exhibited that the LOD was 0.99 μg/L, the LOQ was 3.29 μg/L, the accuracy (spike and recovery experience) was 87.67%?106.27%, and the intra-batch and inter-batch imprecision expressed as coefficient of variation ( CV) was 2.8%?7.3%, and the healthy reference range was 34.31-71.79 μg/L. The serum NCC levels in the WD group were 102.39 (74.38, 144.04) μg/L, which was significantly higher than those in healthy group (51.45±10.34) μg/L ( Z=?7.967, P<0.01). Conclusions:The established IP-ICP-MS method for detecting serum NCC meets the required analytical performance criteria. It is simply operative, highly sensitive, and provides accurate and reliable results, which could be used for clinical detection.

Objective:To establish a method for detecting serum non-ceruloplasmin-bound copper (NCC) by immunoprecipitation-inductively coupled plasma mass spectrometry (IP-ICP-MS) and evaluate its clinical application.Methods:Methodological evaluation research. Immunoprecipitation was first used to separate serum ceruloplasmin, followed by detection of serum NCC levels using ICP-MS. Two levels of quality control serum were reconstituted to determine the limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision of the self-developed method. The serum samples of 131 healthy individuals (healthy group) and 69 first-time diagnosed Wilson′s disease (WD) patients(WD group) from November 2023 to June 2024 in the Affiliated Hospital of Institute of Neurology, Anhui University of Traditional Chinese Medicine were collected. Using self-developed method detected the serum NCC levels, established the healthy reference intervals, and NCC levels between the two groups were compared using the Mann-Whitney U test. Results:The calibration curve exhibited excellent lineary across the concentration range of 1.562 to 300.000 μg/L, as demonstrated by coefficient of determination ( R2) of 0.999. The self-developed NCC method exhibited that the LOD was 0.99 μg/L, the LOQ was 3.29 μg/L, the accuracy (spike and recovery experience) was 87.67%?106.27%, and the intra-batch and inter-batch imprecision expressed as coefficient of variation ( CV) was 2.8%?7.3%, and the healthy reference range was 34.31-71.79 μg/L. The serum NCC levels in the WD group were 102.39 (74.38, 144.04) μg/L, which was significantly higher than those in healthy group (51.45±10.34) μg/L ( Z=?7.967, P<0.01). Conclusions:The established IP-ICP-MS method for detecting serum NCC meets the required analytical performance criteria. It is simply operative, highly sensitive, and provides accurate and reliable results, which could be used for clinical detection.

Objective To analyze the changes of liver and kidney function, blood glucose and lipid metabolism at different follow-up time points of different treatment regimens, and to provide reference for clinical optimization and adjustment of medication in HIV/AIDS patients. Methods The changes of liver and kidney function, blood glucose and lipid metabolism at seven follow-up time points were analyzed retrospectively. The baseline blood collection time of HIV /AIDS patients was set as the starting point, and the final follow-up time was set as the end point. The seven follow-up points were 0, 3, 6, 9, 12, 18 and 24 months respectively. Results There were statistically significant differences in the distribution of sex, age, education, marital status, WHO staging, infection route, and baseline CD4+T lymphocyte count among 605 enrolled patients based on different treatment regimens. Liver function: The level of T-Bil in group E was higher than that of baseline at 9M, 12M, 18M and 24M after treatment (P<0.01); In group F, the level of T-Bil was higher than that of baseline at 9M after treatment (P=0.001); The levels of ALT in group C at the six follow-up points after treatment were higher than the baseline (P<0.001); The level of AST in group C was higher than that of baseline after 3M and 6M treatment (P<0.05). Renal function: The level of UREA in group C was higher than that in baseline after 6M treatment (P=0.007); The level of UREA in group F was higher than that in the baseline after 12M treatment (P<0.001); The level of UA in group F was higher than that of baseline after 3M, 6M and 12M treatment (P<0.05). Blood lipid and blood glucose: The levels of Glu at some follow-up points after ART treatment in group A and group C were higher than that at baseline (P<0.05); The levels of TG at some follow-up points in group A, group E and group F after ART treatment were higher than those at baseline (P<0.05); The levels of TC at some follow-up points in group A, group B, group C, group E and group F after ART treatment were all higher than the baseline (P<0.05). Conclusion Regular monitoring of changes in laboratory indicators of different treatment regimens during ART is of great importance to the prognosis of patients. Different laboratory indicators should be monitored according to different treatment regimens to effectively prevent adverse reactions caused by different treatment regimens.

Duchenne muscular dystrophy(DMD)is a lethal,progressive,X-linked recessive hereditary muscle disease caused by a mutation in the gene encoding dystrophin.Currently,no cure for DMD is available,and clinical research is progressing slowly.The establishment of animal models is becoming increasingly important for experimental research on DMD.Following the research findings that mdx mice have the same pathogenesis as DMD patients,this model is widely used in the study of DMD pathogenesis and drug development.Mitochondrial injury is one of the most important pathological mechanisms of DMD,and mitochondrial protection is a potential therapeutic strategy for DMD,and thus it is significant to study mitochondrial injury in mdx mice.This article reviews the progress of research into mitochondrial injury in DMD model mdx mice in recent years to provide a reference for related experiments.

Purpose To investigate the clinical value of multi-center digital mammography radiomics nomogram model in predicting triple-negative breast cancer(TNBC).Materials and Methods The digital mammograms of 462 patients with pathologically confirmed breast cancer from November 2016 to March 2022 were retrospectively analyzed,including 243 cases from Yijishan Hospital of Wannan Medical College(institution 1),106 cases from Fuyang People's Hospital(institution 2)and 113 cases from Taihe People's Hospital(institution 3).According to the results of immunohistochemistry,a total of 349 breast cancer patients in institution 1 and institution 2 were randomly divided into the training group(244 cases,including 41 TNBC and 203 non-TNBC)and the validation group(105 cases,including 18 TNBC and 87 non-TNBC)according to the ratio of 7∶3,113 breast cancer patients(24 TNBC and 89 non-TNBC)from institution 3 were included in the external validation group.Comparing the mediolateral oblique and cranial cauda digital mammography images,the mammography imaging with larger lesion areas were selected,and the image segmentation and radiomics feature extraction were performed.The radiomics model was constructed by using Logistic regression.The clinicopathological parameters and radiomics scores were used to construct a nomogram.Receiver operating characteristic and decision curve analysis were used to evaluate the model performance.To compare The predictive performance between the models was compared.Results Finally,four radiomics features closely related to TNBC were selected to construct an radiomics model.The area under the curve,sensitivity and specificity of TNBC predicted by the radiomics model in training group,validation group and external test group were 0.868,90.24%and 72.91%,0.827,72.22%and 75.86%,0.837,70.83%and 78.65%,respectively.The area under the curve,sensitivity and specificity of TNBC predicted by the combined model in the training group,validation group and external test group were 0.903,80.49%and 86.70%,0.890,77.78%and 88.51%,0.870,62.50%and 85.39%,respectively.The combined model was better than the single image omics model in predicting TNBC,and the difference was statistically significant between the training group and the verification group(Z=2.061,2.064,both P<0.05),but not between the external test group(Z=1.223,P=0.221).In three group,decision curve analysis showed that the nomogram predicted a higher net benefit than the radiomics model for triple-negative breast cancer.Conclusion The radiomics model has high diagnostic efficiency in predicting TNBC,and the nomogram model combined with the radiomics score and histological grading can further improve the prediction efficiency.

Parkinson's disease （PD） is a progressive chronic neurodegenerative disorder with a complex pathogenesis involving oxidative stress， neuroinflammation， mitochondrial dysfunction， and other factors. Currently， the clinical treatment of PD mainly includes levodopa， dopamine receptor agonists， monoamine oxidase B inhibitors， catechol-O-methyltransferase inhibitors， and anticholinergic drugs， but there is a lack of disease-modif g therapies that can definitively improve disease progression. According to the understanding of traditional Chinese medicine （TCM）， PD is characterized by asthenia in origin and sthenia in superficiality. It is primarily caused by liver-kidney Yin deficiency， Qi-blood insufficiency， and closely related to wind， fire， phlegm， and blood stasis. Numerous clinical practices have shown that TCM has significant clinical value in the prevention and treatment of PD， the management of motor and non-motor symptoms， and the neuroprotection of dopaminergic neurons. The underlying mechanisms of TCM include antioxidative stress， anti-neuroinflammation， and regulation of mitochondrial dysfunction. This article categorized and summarized the pathogenesis of PD， systematically elucidated the pharmacological actions and molecular mechanisms of TCM monomer extracts and compounds in the prevention and treatment of PD， and provided the latest clinical research progress， aiming to provide references for the development and clinical use of TCM for PD.

Objective:To explore the relationship between drug resistance occurrence and the distribution pattern of polymorphic loci in individuals with human immunodeficiency virus infection/acquired immunodeficiency syndrome (HIV/AIDS) treated with highly active anti-retroviral therapy (HAART).Methods:HAART-failed HIV/AIDS patients who successfully amplified the gene sequences of the pol region between June 2015 and December 2021 from 16 prefecture-level administrative regions in Yunnan Province were included.The resistant sequences were classified using the human immunodeficiency virus (HIV) basic local alignment search tool (BLAST) and validated through MEGA 6.0, and the obtained sequences were submitted to the Stanford University HIV Drug Resistance Database to identify drug resistance loci. The distribution of polymorphic loci was analyzed across patients exhibiting varying degrees of drug resistance, different treatment regimens and distinct HIV-1 subtypes.Changes of the frequencies of polymorphic loci in patients with different degrees of drug resistance were analyzed using trend chi-square test. Statistical comparisons and further paired comparisons were performed using chi-square test.Results:Gene sequences were amplified from 1 453 patients, and the resistance testing results showed 954 sensitive, 224 potentially or low resistant, 189 moderately resistant, and 86 highly resistant patients. The frequencies of mutations I15V, L19I, D60E in the HIV-1 protease region (PR region) and E36A, T39D, S48T mutations in the HIV-1 reverse transcriptase region (RT region) showed a decreasing trend as the degree of HIV-1 resistance escalated ( χ2trend=19.86, 9.16, 13.66, 37.64, 18.44 and 40.86, respectively, all P<0.01). Conversely, the mutations V77I in the PR region and K122E in the RT region showed an ascending trend ( χ2trend=12.19 and 10.03, respectively, both P<0.01). Distinct treatment groups, namely zidovudine (AZT)+ lamivudine (3TC)+ lopinavir/ritonavir (LPV/r), AZT+ 3TC+ efavirenz (EFV), AZT+ 3TC+ nevirapine (NVP), and tenofovir (TDF)+ 3TC+ EFV, were examined. Statistically significant differences in the frequencies of mutations E35D, M36I, and D60E in the PR region, as well as S48T, K122E, and R211K in the RT region, were observed among these treatment groups ( χ2=22.46, 9.32, 14.46, 26.85, 18.92 and 24.26, respectively, all P<0.05). In paired comparisons, AZT+ 3TC+ LPV/r group displayed higher frequencies of E35D, M36I, and D60E mutations, the AZT+ 3TC+ EFV group showed a higher frequency of S48T mutation, the AZT+ 3TC+ NVP group showed a higher frequency of K122E mutation, and the TDF+ 3TC+ EFV group exhibited a higher frequency of R211K mutation, all with statistically significant differences (all P<0.008). The differences in the frequencies of T12S, I15V, L19I, M36I, V77I, L89M in the PR region and E53D, I135V, S162C, R211K, K277R in the RT region among circulating recombinant form (CRF)08_BC, CRF07_BC and CRF01_AE subtype group were statistically significant ( χ2=693.60, 712.51, 798.11, 434.85, 386.91, 657.78, 932.58, 409.21, 344.39, 469.44 and 260.48, respectively, all P<0.001). In paired comparisons, the frequencies of T12S, I15V, L19I, E53D, I135V, S162C and R211K in CRF08_BC subtype, the frequencies of V77I and K277R in CRF07_BC subtype, and the frequencies of M36I and L89M in CRF01_AE subtype were higher than those in the other two groups, and the differences were all statistically significant (all P<0.017). Conclusions:The polymorphic loci resulting from HIV-1 HAART failure show different distribution patterns across various degrees of drug resistance, treatment regimens and HIV-1 subtypes.These loci demonstrate both specific and shared characteristics. It is necessary to enhance the surveillance of select polymorphic loci.

ObjectiveTo explore the mechanism of Dendrobium huoshanense polysaccharide （DHP） against inflammatory damage of neurons in Parkinson's disease （PD） model. MethodSH-SY5Y cells were randomized into blank group， model group， and DHP group. The survival rate of cells was measured by thiazole blue（MTT） assay， and the levels of lactate dehydrogenase （LDH）， reactive oxygen species （ROS）， malondialdehyde （MDA）， and superoxide dismutase （SOD） were measured by colorimetric analysis. BV-2 microglia were classified into blank group， model group， DHP group， and MCC950 group （positive control group）， and enzyme-linked immunosorbent assay （ELISA） was applied to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. The expression of NOD-like receptor protein 3 （NLRP3）， adaptor protein apoptosis-associated dot protein （ASC）， cysteine aspartic protease-1 （Caspase-1）， and IL-1β was measured by Western blot. A total of 50 C57BL/6 mice were randomized into blank group， model group， DHP low-dose （100 mg·kg^-1） group， DHP equivalent-dose （350 mg·kg^-1） group， and MCC950 group （positive control group）， 10 mice in each group. The motor balance and coordination of C57BL/6 mice were observed by beam walking test， tail suspension test and rotarod test. The levels of Iba-1 and tyrosine hydroxylase （TH） were detected by immunofluorescence staining. The damage of dopaminergic neurons in the substantia nigra was detected by FJB staining. The levels of inflammatory factors such as IL-1β， IL-18， and TNF-α in mouse midbrain tissues were detected by ELISA and the protein levels of NLRP3， ASC， Caspase-1， and IL-1β protein were measured by Western blot. ResultCompared with the blank group， the SH-SY5Y model group showed decreased cell survival， increased levels of LDH， ROS， and MDA （P<0.05）， and decreased levels of SOD （P<0.05）. Compared with the model group， the DHP group demonstrated increased cell survival， decreased levels of LDH， ROS， and MDA （P<0.01）， and increased level of SOD （P<0.01）. Compared with the blank group， BV-2 model group had high levels of IL-1β， IL-18， and TNF-α （P<0.05） and high protein expression of NLRP3， Caspase-1， IL-1β， and ASC （P<0.05）. Compared with the model group， DHP and MCC950 groups demonstrated low levels of IL-1β， IL-18， and TNF-α （P<0.01） and low protein expression of NLRP3， Caspase-1， IL-1β， and ASC （P<0.01）. Compared with the blank group， the C57BL/6 model group displayed long time to pass the balance wood （P<0.05）， short time spent on the rod in the rotarod test （P<0.05）， high levels of IL-1β， IL-18， and TNF-α （P<0.05） and expression of Iba-1 in the midbrain substantia nigra （P<0.05）， low TH expression （P<0.05）， more positive neurons in the FJB staining （P<0.05）， and high expression of NLRP3， Caspase-1， ASC， and IL-1β proteins （P < 0.05）. Compared with the model group， the mice in the DHP and MCC950 groups had short time to pass the balance beam （P<0.01）， long time spent on the rod （P<0.01）， low levels of IL-1β， IL-18， and TNF-α （P<0.01）， low Iba-1 expression in midbrain substantia nigra （P<0.01）， high TH expression （P<0.01）， and small number of positive neurons in the midbrain substantia nigra （P<0.01）. The expression of NLRP3， ASC， and IL-1β proteins was lower in the MCC950 group （P<0.01）， and the expression of NLRP3， ASC， Caspase-1 and IL-1β proteins was lower in the DHP equivalent-dose group （P<0.01） than in the model group. ConclusionDHP has anti-oxidative stress effect. It regulates the expression of NLRP3 inflammasome and inhibits the overactivation of microglia， thereby alleviating the neuroinflammatory injury in PD and exerting the neuroprotective effect.