AIM:This study aims to investigate the mechanism by which Qingfei-Jiedu-Huatan Formula(QJHF)regulates autophagy in alveolar macrophages through mTOR in the treatment of severe pneumonia(SP)in rats.METHODS:Sixty SPF-grade male rats were randomly assigned to six groups:control,model,QJHF,moxifloxacin(MOX),rapamycin(RAPA),and QJHF+RAPA,with ten rats in each group.An SP rat model was established using Klebsiella pneumoniae.After seven days of treatment,changes in IL-33 and IFN-γ levels in bronchoalveolar lavage fluid(BALF)were measured using ELISA.Histopathological alterations in lung tissue were assessed via HE staining,and the autophagy of alveolar macrophages was detected using immunofluorescence co-localization methods.The expression levels of mTOR,beclin-1,and LC3 mRNA in lung tissue were analyzed using qPCR,while Western blot was employed to assess the protein levels of p-mTOR/mTOR,beclin-1,and LC3-II/LC3-I.RESULTS:Compared to the control group,the model group exhibited a deteriorated condition,characterized by alveolar wall rupture and thickening,significant inflammatory cell infiltration in the alveolar cavity,and extensive lung tissue damage(P<0.01).Elevated levels of IL-33 and IFN-γ in BALF were also observed(P<0.01),along with increased colocalization of CD68 and LC3 in immunofluorescence analy-sis.The mTOR mRNA expression in lung tissue decreased(P<0.01),while LC3 and beclin-1 mRNA expressions in-creased(P<0.01).Additionally,the protein expression ratio of p-mTOR/mTOR decreased(P<0.01),whereas LC3-II/LC3-I and beclin-1 protein levels increased(P<0.01).In comparison to the model group,significant improvements were noted after treatment with QJHF and MOX(P<0.01),while RAPA treatment led to a worsening of these indicators(P<0.05).A slight improvement was observed with the QJHF combined with RAPA intervention,though this was not statisti-cally significant.No significant differences were found between the MOX and QJHF groups.However,the QJHF+RAPA group displayed notable improvements in various indicators compared to the RAPA group(P<0.05).CONCLUSION:The QJHF can mitigate the inflammatory response associated with severe pneumonia,potentially by activating mTOR phos-phorylation activity,which in turn inhibits excessive autophagy in alveolar macrophages.

Objective To investigate the effects of different cultivation method and harvest periods on chromatic characteristics and the primary bioactive components of Dendrobium officinale.Methods Fifteen batches of three-year-old Dendrobium officinale were categorized into five cultivation groups:DP-ZP,YB-ZP,HS-ZP,SP-ZP,and ZJ-ZP,with three batches per group.Thirty-six batches of Dendrobium officinale samples were categorized into twelve harvest groups according to different harvest periods and growth years:the T1 to T6 groups were three-year-old samples,and the F1 to F6 groups were four-year-old samples.Chromaticity values of Dendrobium officinale under different cultivation methods and harvest periods were measured using colorimetry.Polysaccharide content under different cultivation method and harvest periods was quantified via ultraviolet spectrophotometry.Mannose,glucose,flavonoids,and phenolic acids(rutin,quercetin,naringenin,and syringic acid)were analyzed in each group under different cultivation methods and harvest periods using HPLC.The total amount of mannose and glucose,the peak area ratio of mannose to glucose,and the total amount of flavonoids and phenolic acids were calculated.The entropy weight-TOPSIS method was applied to assign objective weights to indicators,followed by the calculation of relative closeness(C)for comprehensive quality assessment of medicinal materials.Results Under different cultivation methods,compared with the DP-ZP group,the b? value of the ZJ-ZP and YB-ZP groups decreased,whereas that of the SP-ZP group increased.The L? value and polysaccharide,syringic acid,naringenin,and quercetin contents decreased,whereas the a? value increased in the YB-ZP,SP-ZP,HS-ZP,and ZJ-ZP groups.The mannose contents decreased in the HS-ZP and SP-ZP groups and increased in the ZJ-ZP group(P<0.05).The glucose content in the ZJ-ZP and SP-ZP groups decreased,the peak area ratio of mannose to glucose increased(P<0.05).Compared with the SP-ZP group,the mannose and glucose contents and the total amount of mannose and glucose were significantly increased,and the peak area ratio of mannose to glucose was decreased in the DP-ZP,YB-ZP,HS-ZP,and ZJ-ZP groups(P<0.01).Compared with the ZJ-ZP group,the rutin contents and the total amount of flavonoids and phenolic acids were decreased in the SP-ZP,HS-ZP,DP-ZP,and YB-ZP groups(P<0.05).Compared with the T3 group,the L? value of the T1,T2,T5,and T6 groups decreased,whereas the b? value increased.The a? value and the peak area ratio of mannose to glucose increased in the T1,T2,and T4 to T6 groups,whereas the polysaccharide and glucose contents and the total amount of mannose and glucose decreased(P<0.05).Compared with the T5 group,the mannose content in the T1 to T4 and T6 groups decreased,whereas the rutin content increased in the T1 to T4 groups(P<0.05).Compared with the T6 group,the syringic acid and quercetin contents increased in the T1 to T5 groups,and the total amount of flavonoids and phenolic acids increased in the T1 to T4 group(P<0.05).Compared with the F1 group,the syringic acid content in the F2 to F6 group increased,the quercetin content decreased(P<0.01).Compared with the F2 group,the glucose and rutin content and the total amount of flavonoids and phenolic acids decreased in the F1 and F3 to F6 groups(P<0.01).Compared with the F3 group,the L? value decreased,the b? value increased,mannose content and the total amount of mannose and glucose decreased in the F1,F2,and F4 to F6 groups.Polysaccharide content in the F1 and F4 to F6 groups decreased(P<0.05).Compared with the F6 group,the peak area ratio of mannose to glucose was significantly decreased in the F1 to F4 groups(P<0.01).Entropy weight-TOPSIS analysis showed that the C of chromatic characteristics and primary bioactive components in DP-ZP(C=0.661 1)and F5(C=0.538 9)groups were the highest.Conclusion Dendrobium officinale cultivated under greenhouse conditions and four-year-old Dendrobium officinale harvested in February exhibit optimal overall quality.These findings provide an experimental basis for the selection of cultivation method and the optimal harvest period of high-quality Dendrobium officinale.

Objective To investigate the effects of different cultivation method and harvest periods on chromatic characteristics and the primary bioactive components of Dendrobium officinale.Methods Fifteen batches of three-year-old Dendrobium officinale were categorized into five cultivation groups:DP-ZP,YB-ZP,HS-ZP,SP-ZP,and ZJ-ZP,with three batches per group.Thirty-six batches of Dendrobium officinale samples were categorized into twelve harvest groups according to different harvest periods and growth years:the T1 to T6 groups were three-year-old samples,and the F1 to F6 groups were four-year-old samples.Chromaticity values of Dendrobium officinale under different cultivation methods and harvest periods were measured using colorimetry.Polysaccharide content under different cultivation method and harvest periods was quantified via ultraviolet spectrophotometry.Mannose,glucose,flavonoids,and phenolic acids(rutin,quercetin,naringenin,and syringic acid)were analyzed in each group under different cultivation methods and harvest periods using HPLC.The total amount of mannose and glucose,the peak area ratio of mannose to glucose,and the total amount of flavonoids and phenolic acids were calculated.The entropy weight-TOPSIS method was applied to assign objective weights to indicators,followed by the calculation of relative closeness(C)for comprehensive quality assessment of medicinal materials.Results Under different cultivation methods,compared with the DP-ZP group,the b? value of the ZJ-ZP and YB-ZP groups decreased,whereas that of the SP-ZP group increased.The L? value and polysaccharide,syringic acid,naringenin,and quercetin contents decreased,whereas the a? value increased in the YB-ZP,SP-ZP,HS-ZP,and ZJ-ZP groups.The mannose contents decreased in the HS-ZP and SP-ZP groups and increased in the ZJ-ZP group(P<0.05).The glucose content in the ZJ-ZP and SP-ZP groups decreased,the peak area ratio of mannose to glucose increased(P<0.05).Compared with the SP-ZP group,the mannose and glucose contents and the total amount of mannose and glucose were significantly increased,and the peak area ratio of mannose to glucose was decreased in the DP-ZP,YB-ZP,HS-ZP,and ZJ-ZP groups(P<0.01).Compared with the ZJ-ZP group,the rutin contents and the total amount of flavonoids and phenolic acids were decreased in the SP-ZP,HS-ZP,DP-ZP,and YB-ZP groups(P<0.05).Compared with the T3 group,the L? value of the T1,T2,T5,and T6 groups decreased,whereas the b? value increased.The a? value and the peak area ratio of mannose to glucose increased in the T1,T2,and T4 to T6 groups,whereas the polysaccharide and glucose contents and the total amount of mannose and glucose decreased(P<0.05).Compared with the T5 group,the mannose content in the T1 to T4 and T6 groups decreased,whereas the rutin content increased in the T1 to T4 groups(P<0.05).Compared with the T6 group,the syringic acid and quercetin contents increased in the T1 to T5 groups,and the total amount of flavonoids and phenolic acids increased in the T1 to T4 group(P<0.05).Compared with the F1 group,the syringic acid content in the F2 to F6 group increased,the quercetin content decreased(P<0.01).Compared with the F2 group,the glucose and rutin content and the total amount of flavonoids and phenolic acids decreased in the F1 and F3 to F6 groups(P<0.01).Compared with the F3 group,the L? value decreased,the b? value increased,mannose content and the total amount of mannose and glucose decreased in the F1,F2,and F4 to F6 groups.Polysaccharide content in the F1 and F4 to F6 groups decreased(P<0.05).Compared with the F6 group,the peak area ratio of mannose to glucose was significantly decreased in the F1 to F4 groups(P<0.01).Entropy weight-TOPSIS analysis showed that the C of chromatic characteristics and primary bioactive components in DP-ZP(C=0.661 1)and F5(C=0.538 9)groups were the highest.Conclusion Dendrobium officinale cultivated under greenhouse conditions and four-year-old Dendrobium officinale harvested in February exhibit optimal overall quality.These findings provide an experimental basis for the selection of cultivation method and the optimal harvest period of high-quality Dendrobium officinale.

Objective·To investigate the effects and molecular mechanisms of phosphatidylethanolamine(PE)on macrophage senescence and its senescence-associated secretory phenotype(SASP),as well as its pathophysiological role in liver injury.Methods·A macrophage senescence model was established using doxorubicin(DOX),followed by PE treatment.A mouse liver injury model was generated via intraperitoneal co-administration of PE and lipopolysaccharide(LPS)to investigate the effects of PE on liver injury.Senescence markers and SASP factors,including senescence-associated β-galactosidase(SA-β-gal),cell cycle inhibitor p21,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6),were evaluated using SA-β-gal staining,quantitative real-time PCR,and Western blotting.RNA sequencing(RNA-seq)was performed,followed by Gene Ontology(GO)cellular component enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,Gene Set Variation Analysis(GSVA),and Gene Set Enrichment Analysis(GSEA),to explore the molecular mechanisms and signaling pathways by which PE promotes macrophage senescence.The expression of endoplasmic reticulum(ER)stress-related proteins,including inositol-requiring enzyme 1 α(IRE1α),spliced X-box binding protein 1(XBP1s),activating transcription factor 6(ATF6),ATF4,and C/EBP homologous protein(CHOP),was analyzed through in vivo and in vitro experiments.Results·PE significantly promoted the expression of senescence markers SA-β-gal,p21,p16 and SASP factors.RNA-seq analysis revealed that ER stress was involved in PE-induced promotion of SASP.Further experiments demonstrated that PE activated the ER stress signaling pathway,promoting macrophage senescence and the expression of SASP factors.In vivo experiments further confirmed that PE exacerbated LPS-induced liver injury in mice through ER stress.Conclusion·PE promotes macrophage senescence and the expression of SASP factors by activating ER stress signaling pathway,thereby aggravating LPS-induced liver injury.

AIM:This study aims to investigate the mechanism by which Qingfei-Jiedu-Huatan Formula(QJHF)regulates autophagy in alveolar macrophages through mTOR in the treatment of severe pneumonia(SP)in rats.METHODS:Sixty SPF-grade male rats were randomly assigned to six groups:control,model,QJHF,moxifloxacin(MOX),rapamycin(RAPA),and QJHF+RAPA,with ten rats in each group.An SP rat model was established using Klebsiella pneumoniae.After seven days of treatment,changes in IL-33 and IFN-γ levels in bronchoalveolar lavage fluid(BALF)were measured using ELISA.Histopathological alterations in lung tissue were assessed via HE staining,and the autophagy of alveolar macrophages was detected using immunofluorescence co-localization methods.The expression levels of mTOR,beclin-1,and LC3 mRNA in lung tissue were analyzed using qPCR,while Western blot was employed to assess the protein levels of p-mTOR/mTOR,beclin-1,and LC3-II/LC3-I.RESULTS:Compared to the control group,the model group exhibited a deteriorated condition,characterized by alveolar wall rupture and thickening,significant inflammatory cell infiltration in the alveolar cavity,and extensive lung tissue damage(P<0.01).Elevated levels of IL-33 and IFN-γ in BALF were also observed(P<0.01),along with increased colocalization of CD68 and LC3 in immunofluorescence analy-sis.The mTOR mRNA expression in lung tissue decreased(P<0.01),while LC3 and beclin-1 mRNA expressions in-creased(P<0.01).Additionally,the protein expression ratio of p-mTOR/mTOR decreased(P<0.01),whereas LC3-II/LC3-I and beclin-1 protein levels increased(P<0.01).In comparison to the model group,significant improvements were noted after treatment with QJHF and MOX(P<0.01),while RAPA treatment led to a worsening of these indicators(P<0.05).A slight improvement was observed with the QJHF combined with RAPA intervention,though this was not statisti-cally significant.No significant differences were found between the MOX and QJHF groups.However,the QJHF+RAPA group displayed notable improvements in various indicators compared to the RAPA group(P<0.05).CONCLUSION:The QJHF can mitigate the inflammatory response associated with severe pneumonia,potentially by activating mTOR phos-phorylation activity,which in turn inhibits excessive autophagy in alveolar macrophages.

Objective:To summarize and analyze the clinical characteristics of systemic lupus erythematosus (SLE) combined with ischaemic stroke and the factors associated with poor prognosis.Methods:A total of 50 patients with SLE combined with ischaemic stroke in the First Affiliated Hospital of Xi′an Jiaotong University from January 2014 to June 2024 were included in the study, the clinical data of the patients were retrospectively collected and summarized, the Shapiro-Wilk test was used to assess the normality of data, and the factors related to poor prognosis were analyzed by logistic regression analyses.Results:Fifty patients with SLE combined with ischaemic stroke had a mean age of (47.1±15.5)years, 80.0%(40/50) were female, the duration of SLE was (5.6±6.3)years, the mean SLEDAI-2K score was (14.3±4.1), the rate of anticardiolipin antibody positivity was 30.0%(15/50), and the rate of β 2-glycoprotein Ⅰ antibody positivity was 28.0%(14/50). The most common clinical manifestations of stroke were impaired limb movement (34.0%) (17/50), cerebral infarction mainly in the cerebral hemisphere (82.0%)(41/50), combined with cerebral haemorrhage in 6.0%(3/50), cerebral leukoencephalopathy in 26.0%(13/50), and cerebral atrophy in 24.0%(12/50). In terms of treatment, the most used immunosuppressant was cyclophosphamide (34.0%, 17/50), 64.0%(32/50) of patients received aspirin, 32.0%(16/50) received clopidogrel and 14.0%(7/50) received anticoagulation. Four deaths and 12 cases of severe disability were found in 50 patients at follow-up, and SLEDAI-2000 scores were positively correlated with the above poor prognosis using univariate [ OR(95% CI)=1.407(1.123,1.764), P=0.003] and multivariate [ OR(95% CI)=1.388(1.097, 1.756), P=0.006] regression analyses. Conclusion:Patients with SLE combined with ischaemic stroke had high disease activity in SLE, and SLEDAI-2000 scores were positively associated with poor prognosis of death and severe disability.

Objective To explore the expression of epidermal growth factor and nuclear transcrip-tion factor kappa B(NF-κB)in elderly gliomas and their roles in malignant progression of the dis-ease.Methods A total of 240 elderly glioma patients undergoing surgical resection in our hospital from January 2020 to January 2023 were enrolled and served as an observation group.Another 100 patients receiving surgical treatment due to traumatic brain injury in our hospital during the same period were recruited and served as the control group.Immunohistochemical assay was used to detect the expression of epidermal growth factor and NF-κB in brain tissue.Cox regression analy-sis was employed to determine the independent influencing factors of malignant progression in the elderly glioma patients.Results The observation group had significantly higher epidermal growth factor expression score and NF-κB expression score than the control group(7.44±1.16 vs 3.68±0.51,6.49±1.02 vs 3.56±0.64,P＜0.01).The age,tumor pathological grade,surgical approach,postoperative radiotherapy,and epidermal growth factor and NF-κB expression levels were inde-pendent factors affecting the progression of elderly glioma patients(P＜0.05,P＜0.01).Conclusion The epidermal growth factor and NF-κB expression levels are independent influen-cing factors of progressive disease in elderly glioma patients,and can serve as evaluation indicators for postoperative progression in the patients.

Objective·To investigate the effects and molecular mechanisms of phosphatidylethanolamine(PE)on macrophage senescence and its senescence-associated secretory phenotype(SASP),as well as its pathophysiological role in liver injury.Methods·A macrophage senescence model was established using doxorubicin(DOX),followed by PE treatment.A mouse liver injury model was generated via intraperitoneal co-administration of PE and lipopolysaccharide(LPS)to investigate the effects of PE on liver injury.Senescence markers and SASP factors,including senescence-associated β-galactosidase(SA-β-gal),cell cycle inhibitor p21,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6),were evaluated using SA-β-gal staining,quantitative real-time PCR,and Western blotting.RNA sequencing(RNA-seq)was performed,followed by Gene Ontology(GO)cellular component enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis,Gene Set Variation Analysis(GSVA),and Gene Set Enrichment Analysis(GSEA),to explore the molecular mechanisms and signaling pathways by which PE promotes macrophage senescence.The expression of endoplasmic reticulum(ER)stress-related proteins,including inositol-requiring enzyme 1 α(IRE1α),spliced X-box binding protein 1(XBP1s),activating transcription factor 6(ATF6),ATF4,and C/EBP homologous protein(CHOP),was analyzed through in vivo and in vitro experiments.Results·PE significantly promoted the expression of senescence markers SA-β-gal,p21,p16 and SASP factors.RNA-seq analysis revealed that ER stress was involved in PE-induced promotion of SASP.Further experiments demonstrated that PE activated the ER stress signaling pathway,promoting macrophage senescence and the expression of SASP factors.In vivo experiments further confirmed that PE exacerbated LPS-induced liver injury in mice through ER stress.Conclusion·PE promotes macrophage senescence and the expression of SASP factors by activating ER stress signaling pathway,thereby aggravating LPS-induced liver injury.

This study identified the types and pathogen carrying status of fleas on the surface of sheep in some areas of southern Xinjiang,and analyzed the genetic evolution differences with respect to related pathogens.The aim was to provide a reference for the local prevention and control of fleas and insect borne infectious diseases.A total of 1 586 fleas were collected from agricultural and pas-toral areas of Tumushuke City and Hotan Prefecture.Flea species were identified on the basis of morphology and the Pulex irritans mi-tochondrial COII gene.Flea DNA was extracted,and PCR was conducted to amplify the Bartonella gltA gene;Arsenophonus,Ana-plasma,Ehrlichia,and Wolbachia 16S rRNA genes;RickettsiaOmpA,17kDa,16S rRNA genes,and Yersinia pestis 16S rDNA gene.The amplified products were sequenced,and the homology of the genes of the three detected pathogens(gltA gene of Bartonella,16S rRNA gene of Wolbachia,and Anaplasma phagocytophilum)with respect to known corresponding genes of the same pathogen in Gen-Bank was analyzed.Phylogenetic trees were constructed with the adjacency method in MEGA 11.0.According to morphological and mo-lecular biology identification results,all fleas collected in this study were Pulex irritans.PCR indicated that the target gene fragments had been added to the mitochondrial COII,BartonellagltA,Wolbachia,and autophagosomal 16S rRNA genes of human fleas,all of which were consistent with the expected fragment sizes.Target bands were not amplified from Ehrlichia,Arsenophonus,spotted fever group Rickettsia,and Yersinia pestis.According to homology and genetic evolution analysis of human flea mitochondrial COII and the corresponding genes of the above-described pathogens,the COX2 gene(ON455234.1)of human fleas in Tumushuke city and Iran ob-tained in this study showed the highest homology(99.84%).The COII gene(NC_063709.1)of human fleas in Hetan City and Hunan region showed the highest homology(100%).Our findings further confirmed that the flea species was Pulex irritans.The PCR amplifi-cation results indicated that the collected Pulex irritans carried multiple pathogens,among which Bartonella and Wolbachia had the highest infection rates,and the infection rate with Anaplasma phagocytophilum was relatively low.This study is the first to discover flea species on the surface of sheep in some areas of southern Xinjiang.Our findings preliminarily confirmed that Bartonella,Wolba-chia,and Anaplasma phagocytophilum are the main Pulex irritans pathogens.

Objective:To explore the application effect of intelligent precision feeding guidance mode in postoperative feeding of children with congenital heart disease, in order to provide a scientific, dynamic and effective feeding management method for postoperative children with congenital heart disease.Methods:A prospective, controlled study design was adopted to select 144 pairs of congenital heart disease patients and their primary caregivers who visited the Pediatric Heart Center of Beijing Anzhen Hospital affiliated to Capital Medical University from March to December 2023 using convenience sampling method. They were randomly divided into a control group and an experimental group, with 72 pairs in each group, using a random number table method. The control group received routine feeding guidance, while the experimental group received intelligent precision feeding guidance mode intervention on the basis of the control group. Compared the infant and child feeding index (ICFI), malnutrition and physical development status, and primary caregiver′s Positive Aspects of Caregiving (PAC) scores between two groups of children with congenital heart disease.Results:Finally, 130 pairs of children and their primary caregivers completed this study, with 64 pairs in the control group and 66 pairs in the experimental group. There were 30 males and 34 females in the control group, with an aged of (11.64 ± 5.03) months; there were 35 males and 31 females in the experimental group, with an aged of (11.03 ± 5.07) months. Among the primary caregivers of the control group, there were 53 mothers and 11 fathers/grandparents/other caregivers; among the main caregivers of the experimental group, there were 62 mothers and 4 fathers/grandparents/other caregivers. The ICFI of the experimental group at 3 and 6 months after surgery were 13.0(10.0, 14.0) and 15.0(13.0, 15.0) points, respectively, which were higher than those of the control group at 11.0(9.0, 13.0) and 12.0(9.0, 14.0) points, and the differences were statistically significant ( Z=8.23, 22.32, both P<0.05). The ICFI qualification rates of the experimental group at 3 and 6 months after surgery were 37.1% (23/62) and 59.3% (35/59), respectively, which were higher than the control group′s 19.7% (12/61) and 31.6% (18/57), and the differences were statistically significant ( χ2=4.59, 8.99, both P<0.05). At 6 months after surgery, the body weight of the experimental group increased was (1.78 ± 0.46) kg, which was higher than that of the control group (1.50 ± 0.52) kg, and the difference was statistically significant ( t=-3.23, P<0.05). In addition, at 6 months after surgery, the incidence of acute malnutrition in the experimental group was 12.1% (8/66), which was lower than that of the control group 25.0% (16/64), and the difference was statistically significant ( χ2=4.36, P<0.05). At 6 months after surgery, the scores of self affirmation, life outlook dimension and the total PAC score of the primary caregiver in the experimental group were (23.45 ± 3.57), (11.97 ± 1.91), and (35.96 ± 4.92) points, respectively, which were higher than those in the control group (22.11 ± 3.40), (11.02 ± 1.90) and (33.13 ± 4.86) points, and the differences were statistically significant ( t=-2.20, -2.86, -2.68, all P<0.05). Conclusions:The use of intelligent precision feeding guidance mode for feeding management of postoperative children with congenital heart disease can effectively improve the positive feelings of caregivers, achieve scientific feeding, increase the ICFI and its qualification rate of children, thereby improving their nutritional status and promoting growth and development.