Objective To establish a cellular model of Parkinson disease by treating retinoic acid(RA)-differentiated SH-SY5Y cells with α-synuclein preformed fibrils(α-Syn PFF).Methods SH-SY5Y cells were divided into undifferentiated and RA-differentiated groups.The expression levels of tyrosine hydroxylase(TH),dopamine transporter(DAT),lymphocyte activation gene 3(LAG3),and Nestin proteins in the cells were detected using Western blotting,whereas those of microtubule-associated protein 2(MAP2)and neu-ronal nuclei(NeuN)were detected using immunofluorescence staining.Furthermore,RA-treated SH-SY5Y cells were divided into control and α-Syn PFF groups,and their levels of chromatin condensation were detected using Hoechst 33342 staining.Nitric oxide(NO)levels were measured using a NO assay kit.Additionally,the protein levels of TH,poly(ADP-ribose)(PAR),and poly(ADP-ribose)poly-merase(PARP)in these cells were detected using Western blotting,whereas their expression levels of phosphorylated α-Syn(pS129-α-Syn)and phosphorylated histone H2AX(γH2AX)were detected using immunofluorescence staining.Results Treatment with RA resulted in a reduction in cell body size and the elongation of protrusions in SH-SY5Y cells.The results of Western blotting showed that RA treatment could increase the TH,DAT,and LAG3 levels and decrease the Nestin level in SH-SY5Y cells(P＜0.05).α-Syn PFF treat-ment decreased the TH protein level and increased the PAR,PARP-l,and cleaved PARP-1 levels in differentiated SH-SY5Y cells(P＜0.05).According to the immunofluorescence results,RA treatment increased the expression levels of MAP2 and NeuN in SH-SY5Y cells(P＜0.001).The α-Syn PFF treatment increased the expression levels of γH2AX and pS129-α-Syn in RA-differentiated SH-SY5Y cells(P＜0.01).The Hoechst 33342 staining results showed that α-Syn PFF treatment led to chromatin condensation in the differentiated SH-SY5Y cells(P＜0.001)and increased the NO levels(P＜0.01).Conclusion A cellular model of Parkinson disease can be established by treating RA-differentiated SH-SY5Y cells with α-Syn PFF.

Purpose To explore whether tryptanthrin(TRYP)can inhibit the malignant behavioral ability of glio-ma cells,and to elucidate the specific mechanism of its action.Methods MTT assay was performed to detect the effect of TRYP on the proliferation of glioma cells;Transwell assay was performed to detect the effect of TRYP on the migration and invasion of glioma cells;AnnexinV-FITC/PI apoptosis assay was performed to detect the effect of TRYP on the apoptosis of glioma cells;PI/RNase cell cycle assay was performed to detect the effect of TRYP on the cell cycle distribution of glioma cells;Western blot assay was performed to detect the effect of TRYP on the protein expressions of p-ERK and c-Myc in glioma cells.The effect of TRYP on the proliferation of glioma cells in vivo was verified by con-structing a subcutaneous transplantation tumor model in nude mice,and the effect of TRYP on the apoptotic ability of cells in the transplantation tumor was detected by TUNEL assay.Immunohistochemistry was used to detect the effect of TRYP on the expression of Ki67,BRAF,c-Myc,and p-ERK proteins in transplanted tumor tissues.Results MTT assay showed that TRYP could effectively inhibit the proliferation of glioma cells(P＜0.001).Transwell assay showed that TRYP could inhibit the invasion and migration of glioma cells(P＜0.001).AnnexinV-FITC/PI cell apoptosis as-say showed that TRYP could promote the apoptosis of glioma cells(P＜0.001).The results of PI/RNase cell cycle as-say showed that TRYP was able to promote the G2 phase block of glioma cells(P＜0.001).Western blot results showed that the expression levels of c-Myc and p-ERK proteins in the glioma cells were significantly reduced after TR-YP treatment(P＜0.001).The results of subcutaneous transplantation tumor model in nude mice showed that TRYP could effectively inhibit the growth rate(P＜0.01)and weight(P＜0.05)of transplanted tumor.TUNEL assay showed that TRYP could promote the apoptosis of tumor cells in transplanted tumor(P＜0.001).Immunohistochemis-try results showed that TRYP could effectively inhibit the protein expression of Ki67(P＜0.01),BRAF,c-Myc,and p-ERK(P＜0.001).Conclusion TRYP can inhibit the proliferation,invasion,and migration ability of glioma cells,promote apoptosis of glioma cells,and block the cell cycle of glioma cells.TRYP may inhibit the malignant pro-gression of glioma cells by suppressing the protein expression of BRAF,c-Myc and p-ERK1/2 in the MAPK/ERK sig-naling pathway.

Objective:To investigate the clinical characteristics and survival outcomes of patients with nasopharyngeal carcinoma (NPC) complicated by multiple primary malignancies (MPCs) in a real-world setting.Methods:A retrospective study was performed on 238 NPC patients with MPCs who received radical radiotherapy at Fujian Cancer Hospital between January 1st, 2004 and December 31st, 2023. The primary endpoints were overall survival (OS) and cumulative survival rate. Survival analysis was conducted using the Kaplan-Meier method with log-rank / Breslow tests, and univariate analysis of prognostic factors was performed using the Cox proportional hazards model.Results:A total of 246 primary malignant tumors were identified in 238 patients, involving 12 organ systems and 39 tumor types. The most common coexisting malignancies occurred in the respiratory and intrathoracic organs [25.2% (62/246)], followed by digestive organ malignancies [22.8% (56/246)], malignancies of the lip, oral cavity, and pharynx [22.8% (56/246)], and thyroid and other endocrine gland malignancies [15.4% (38/246)]. The median OS was 186 months, and the 3-, 5-, and 10-year cumulative survival rates were 90.84%, 85.25%, and 69.45%, respectively. Poorer survival was associated with male sex, age>48 years at onset, locally advanced disease (stage IVA), synchronous MPCs and/or digestive system malignancies, fewer total cycles of chemotherapy, and lack of concurrent or adjuvant chemotherapy.Conclusions:In patients with NPC, MPCs most frequently involve the respiratory system, digestive system, and head and neck organs (including the thyroid). Male sex, older age, locally advanced primary NPC, synchronous and/or digestive system MPCs, fewer chemotherapy cycles, and lach of concurrent or adjuvant chemotherapy were significantly associated with poorer prognosis.

Objective: To discover molecular markers associated with lung adenocarcinoma diagnosis/prognosis and drug resistance through screening of differentially expressed genes based on published chip data in gene expression databases using bioinformatics methods. Methods: Comprehensive analysis was performed in available mRNA microarray datasets including lung adenocarcinoma tissues dataset GSE32863 and lung adenocarcinoma taxane-platin resistance dataset GSE77209 from the gene expression omnibus(GEO) database. Gene ontology enrichment analysis, gene pathway enrichment analysis and protein interaction network analysis were performed based on significantly correlated genes. The expression level of genes was validated in the cancer genome atlas(TCGA) dataset. Survival differences were assessed by the log-rank test in TCGA lung adenocarcinoma dataset. Based on the publications genomics of drug sensitivity in cancer(GDSC) database in CellMiner cross database(CellMiner CDB), Pearson correlation analysis was used to analyze the correlation between differentially expressed genes and the half-maximal inhibitory concentration(IC50) of anticancer drugs. Results : There were a total of 77 genes which had a different expression in resistance lung adenocarcinoma cells and lung adenocarcinoma cancer tissues. The functional enrichment analysis showed that these co-different expression genes were mainly enriched in microtubule, extracellular exosome, cell cycle and signaling by nuclear receptors. Protein-protein interactions(PPI) network screened 6 most connected genes as molecular complex(MCODE). Among the MCODE, overexpressed ubiquitin conjugating enzyme E2 T(UBE2T), kinesin family member 20A(KIF20A), PCNA clamp associated factor(KIAA0101), pituitary tumor-transforming gene 1(PTTG1) and NIMA related kinase 2(NEK2) were associated with poor outcomes. Survival analysis results showed that these five genes were upregulated in lung adenocarcinoma tissues and drug-resistant cells and were significantly associated with poor prognosis in lung adenocarcinoma patients. Drug sensitivity analysis results suggested that high expression of PTTG1 and UBE2T was significantly associated with sensitivity to multiple anticancer drugs, including paclitaxel and docetaxel. RT-PCR validation showed that PTTG1 andUBE2T were highly expressed in docetaxel-resistant cells A549-TXR and H358-TXR. Conclusion PTTG1 andUBE2T holds the potential to be chemoresistance markers in lung adenocarcinoma.

Stem cells play a crucial role in maintaining tissue regenerative capacity and homeostasis. However, mechanisms associated with stem cell senescence require further investigation. In this study, we conducted a proteomic analysis of human dental pulp stem cells (HDPSCs) obtained from individuals of various ages. Our findings showed that the expression of NUP62 was decreased in aged HDPSCs. We discovered that NUP62 alleviated senescence-associated phenotypes and enhanced differentiation potential both in vitro and in vivo. Conversely, the knocking down of NUP62 expression aggravated the senescence-associated phenotypes and impaired the proliferation and migration capacity of HDPSCs. Through RNA-sequence and decoding the epigenomic landscapes remodeled induced by NUP62 overexpression, we found that NUP62 helps alleviate senescence in HDPSCs by enhancing the nuclear transport of the transcription factor E2F1. This, in turn, stimulates the transcription of the epigenetic enzyme NSD2. Finally, the overexpression of NUP62 influences the H3K36me2 and H3K36me3 modifications of anti-aging genes (HMGA1, HMGA2, and SIRT6). Our results demonstrated that NUP62 regulates the fate of HDPSCs via NSD2-dependent epigenetic reprogramming.
Humans ; Dental Pulp/cytology* ; Nuclear Pore Complex Proteins/genetics* ; Cellular Senescence/genetics* ; Stem Cells/metabolism* ; Epigenesis, Genetic ; Cell Proliferation ; Cell Differentiation ; Histone-Lysine N-Methyltransferase/metabolism* ; Cells, Cultured ; Cellular Reprogramming ; Cell Movement ; Proteomics

To develop biocontrol agents for the control of kiwifruit bacterial canker, we isolated a strain CXG2-5 with inhibitory activity against Pseudomonas syringae pv. actinidiae (Psa), the pathogen of kiwifruit bacterial canker, from the rhizosphere soil of kiwifruit by the plate confrontation test. The strain was identified by morphological observation, physiological and biochemical tests, and molecular biological methods. The indoor control efficacy of the strain was determined by the inoculation of the strain into detached branches with wounds and into leaf discs by vacuum infiltration. The ability of the strain to expand and colonize leaf veins was determined by fluorescent labeling and scanning electron microscopy. Subsequently, the strain was prepared into microcapsules, the field control efficacy of which was evaluated. The strain CXG2-5 was identified as Pseudomonas benzenivorans. It demonstrated good antagonistic activity against Psa, with an inhibition zone diameter of 22 mm and an inhibition rate of 72.7%. The preventive effects of the strain on kiwifruit bacterial canker were better than the therapeutic effects on both detached branches and leaves, with the preventive effects reaching 65% and 92.4%, respectively. The control effect of microcapsules of this strain in the field reached 60.89%, which was slightly lower than that of 20% kasugamycin and higher than that of Bacillus subtilis wettable powder. In conclusion, strain CXG2-5 serves as a candidate for the control of kiwifruit bacterial canker, and the prepared microcapsules have good value for development and application.
Actinidia/microbiology* ; Plant Diseases/prevention & control* ; Pseudomonas syringae ; Pseudomonas/isolation & purification* ; Capsules ; Antibiosis ; Biological Control Agents ; Pest Control, Biological/methods*

ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis， evaluate its potential clinical application value， and compare the therapeutic effect with that of Cranial Painkiller granules， so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis， and the rats were randomly divided into the normal group， model group， carbamazepine group （60 mg·kg^-1）， Cranial Painkiller granules group （2.70 g·kg^-1）， and low， medium， and high dosage groups of Cranial Painkiller pills' extract powder （1.35， 2.70， 5.40 g·kg^-1） according to the basal mechanical pain thresholds， and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling， and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin （HE） staining was used to detect pathological changes in the trigeminal ganglion， and enzyme-linked immunosorbent assay （ELISA） was used to detect the inflammatory factors interleukin-1 （IL-1）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， and tumor necrosis factor-α （TNF-α） levels in rat serum， as well as neuropeptide substance P （SP） and β-endorphin （β-EP） levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3， ASC， Caspase-1， and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group， the pain threshold of rats in the model group showed a continuous significant decrease （P<0.01）. The pathological damage of brain tissue was significant （P<0.01）， and the inflammatory levels of IL-1， IL-6， IL-8， and TNF-α in serum were significantly elevated （P<0.01）. The level of the SP in the brain tissue was significantly elevated （P<0.01）， and the level of β-EP was significantly reduced （P<0.01）， while the level of OTULIN was significantly reduced， and NLRP3， ASC， and Caspase-1 protein levels were significantly elevated （P<0.01）. After administration of the drug， compared with the model group， the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased （P<0.01）. The inflammatory levels of IL-1， IL-6， IL-8， and TNF-α and SP levels significantly decreased （P<0.01）， and the β-EP levels were significantly elevated （P<0.01）， while the levels of OTULIN protein were significantly elevated （P<0.05， P<0.01）， and the levels of NLRP3， ASC proteins were decreased （P<0.01）in high dose Cranial Painkiller pills' extract powder. Meanwhile， compared with those in the model group， the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement （P<0.05， P<0.01）. ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis， and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.

ObjectiveTo investigate the protective effect and mechanism of verbenalin on mouse mononuclear macrophage leukemia cells （RAW264.7） damaged by human coronavirus （HCoV）-229E infection， thereby providing experimental evidence for its development and application. MethodsRAW264.7 macrophages were infected with different concentrations of HCoV-229E to establish a coronavirus-induced macrophage injury model using the cell counting kit-8 （CCK-8） assay for assessing cell proliferation and viability. Cells were randomly divided into four groups： normal control， verbenalin group （125 μmol·L^-1）， model group （HCoV-229E）， and HCoV-229E + verbenalin group （HCoV-229E + 125 μmol·L^-1 verbenalin）. Cell viability was measured using the CCK-8 assay， and the maximum non-toxic concentration （CC₀）， half-maximal cytotoxic concentration （CC₅₀）， half-maximal effective concentration （EC₅₀）， and selectivity index （SI） of verbenalin were calculated. Calcein/PI double staining was used to assess cell viability and cytotoxicity， and JC-1 staining was applied to evaluate changes in mitochondrial membrane potential （MMP）. mito-Keima adenovirus labeling was used to assess mitophagy levels in each group. ResultsA macrophage infection model was successfully established by infecting RAW264.7 cells with the original concentration of HCoV-229E for 36 h. The CC₀ of verbenalin was 125 μmol·L^-1. The CC₅₀ was 448.25 μmol·L^-1. The EC₅₀ against HCoV-229E-infected cells was 46.28 μmol·L^-1， and the SI was 9.68. Compared with the normal group， the model group showed significantly reduced cell survival rate （P<0.01）， increased cell death rate （P<0.01）， decreased MMP （P<0.01）， and suppressed mitophagy （P<0.01）. In contrast， verbenalin treatment significantly improved cell survival rate （P<0.01）， reduced cell death rate （P<0.01）， alleviated MMP loss （P<0.01）， and enhanced mitophagy levels （P<0.01） compared with the model group. ConclusionVerbenalin can enhance the survival rate of macrophages following HCoV-229E infection. The underlying mechanism may be associated with the activation of mitophagy， maintenance of MMP stability， and alleviation of mitochondrial damage.

ObjectiveThis paper aims to investigate the therapeutic effects of Jidesheng Sheyao tablets on varicella-zoster virus （VZV） and its associated postherpetic neuralgia （PHN） to provide experimental evidence for the clinical application and secondary development of Jidesheng Sheyao tablets. MethodsFifty-six guinea pigs were randomly divided into seven groups according to their body weight， namely the normal group， the model group， the positive control group， the high-dose group， medium-dose group， and low-dose group of Jidesheng Sheyao tablets （1.92， 0.96， 0.48 g·d^-1）， and the group treated with oral administration combined with topical application of Jidesheng Sheyao tablets （0.96 g·d^-1 + 1.2 g·kg^-1·d^-1）. The skin on the back of the guinea pigs in each group was depilated and then abraded with sandpaper. Except for the normal group， 200 μL of VZV solution was dropped on the damaged parts of the back of the guinea pigs in the other groups， and the infection lasted for 2 consecutive days. The drug administration started 2 hours after the infection on the first day and lasted for 7 days. The pathological changes of the back of the guinea pigs in each group were observed every day starting from the second day after the infection. On the 7^th day， the guinea pigs were sacrificed by CO₂ anesthesia. The locally infected skin was taken， and the viral DNA nucleic acid load was detected by real-time polymerase chain reaction （Real-time PCR）. The pathological histology examination was carried out after hematoxylin-eosin （HE） staining. Seventy rats were randomly divided into seven groups according to their body weight， namely the normal group， the model group， the positive control group， the high-dose group， medium-dose group， and low-dose group of Jidesheng Sheyao tablets （1.08， 0.54， 0.27 g·d^-1）， and the group treated with oral administration combined with topical application of Jidesheng Sheyao tablets （0.54 g·d^-1 + 1.2 g·kg^-1·d^-1）. The rats in each group （except the normal group） were subcutaneously inoculated with 50 μL of VZV suspension between the web of the first and second fingers of the left forelimb. The skin on the back of the rats was depilated， and the drug administration started 2 hours after the infection and lasted for 10 days. The mechanical withdrawal threshold of the paws of the rats was detected by a Von Frey filament algometer before inoculation and on the 1^st， 3^rd， 6^th， 8^th， and 10^th days after inoculation， and the thermal withdrawal reflex latency of the paws of the rats was detected by a hot and cold plate algometer. On the 10^th day after the virus inoculation， the rats were anesthetized after the behavioral examination， and the dorsal root ganglia of the spinal cord and spinal cord segments were taken. The contents of substance P （SP）， neurokinin （NK）， tumor necrosis factor-α （TNF-α）， and interleukin-1β （IL-1β） in the dorsal root ganglia of the spinal cord and spinal cord were detected by enzyme-linked immunosorbent assay （ELISA）. ResultsCompared with those in the normal group， the guinea pigs in the model group had obvious skin herpes lesions （P<0.01）. The viral nucleic acid load was high （P<0.01）， and there were disorganized subcutaneous cellular structures and obvious infiltration of inflammatory cells and cell necrosis （P<0.01）. The mechanical withdrawal threshold of the paws and the thermal withdrawal reflex latency of the paws of the rats were significantly decreased （P<0.05）， and the contents of NK， SP， TNF-α， and IL-1β in the dorsal root ganglia of the spinal cord and spinal cord of the rats were significantly increased （P<0.01）. Compared with the model group， the high-dose and medium-dose groups of topical administration of Jidesheng Sheyao tablets and the group of oral administration combined with topical application could significantly improve the lesions such as skin redness and herpes of the guinea pigs caused by VZV infection （P<0.01）， reduce the VZV viral nucleic acid load in the skin tissues of the guinea pigs （P<0.01）， alleviate the degree of inflammatory cell infiltration and skin cell necrosis in the skin tissue （P<0.05）， significantly increase the mechanical withdrawal threshold of the paws and the thermal withdrawal reflex latency of the paws of the rats （P<0.05）， and decrease the contents of NK， SP， TNF-α， and IL-1β in the dorsal root ganglia of the spinal cord and spinal cord of the rats （P<0.01）. ConclusionJidesheng Sheyao tablets demonstrated significant therapeutic effects on VZV-induced skin infections and postherpetic neuralgia （PHN）， providing a promising candidate for the prevention and treatment of VZV infections.

ObjectiveTo construct an animal model of respiratory syncytial virus（RSV）-infected pneumonia suitable for preclinical studies. MethodsThe virulence of RSV to the four cell lines was observed by cytopathic effect （CPE）， and 50% tissue culture infective dose（TCID₅₀） was calculated. Twenty BALB/c mice were randomly divided into a normal group and a model group. Six BALB/c-hIGF1R mice served as the humanized IGF1R model group. Except for the normal group， the other groups received intranasal RSV infection on days 1 and 3 to establish a viral pneumonia model. The efficacy of establishing an RSV-induced pneumonia animal model based on humanized insulin-like growth factor 1 receptor （IGF1R） mice was evaluated by measuring organ indices， peripheral blood lymphocyte percentages， pulmonary pathology and imaging， and pulmonary viral load. Additionally， ten BALB/c mice served as normal group， and thirty-two BALB/c-hIGF1R mice were randomly assigned to humanized IGF1R model group， ribavirin group （82.5 mg·kg^-¹·d^-¹）， and high and low dose groups of Lianhua Qingwen （3.3 mg·kg^-¹·d^-¹ ， 1.65 mg·kg^-¹·d^-¹）， with 8 mice per group. The viral load in lung tissue was measured after ribavirin and Lianhua Qingwen intervention， and the model was applied to the evaluation of anti-RSV drugs. ResultsIn the lungs of the humanized IGF1R model group， large solid and diffuse ground-glass shadows were seen， and the lung volume was significantly increased （P<0.01）. The lung index was significantly increased （P<0.01）， and both the spleen index and thymus index were significantly decreased （P<0.01）. The percentages of CD3⁺ and CD4⁺T cells were significantly decreased （P<0.05）， and there was a large amount of inflammation and stasis in the perivascular area of the lung tissue， which was predominantly characterized by lymphocytes. The endothelium of blood vessels was partially detached， with a small number of eosinophils. After infecting BALB/c-hIGF1R mice with RSV， the expression of viral nucleic acids in the lung tissue of the mice was significantly increased， with significant differences compared with the normal group （P<0.01）. The expression of viral nucleic acids in the ribavirin group and the high and low dose groups of Lianhua Qingwen was significantly reduced， with significant differences compared with the normal group （P<0.01）. ConclusionHumanized IGF1R mice are more susceptible to respiratory SVC， and the animal model of RSV-infected pneumonia based on humanized IGF1R mice was successfully constructed， which is suitable for the evaluation of anti-RSV drugs.