OBJECTIVE: To investigate the biological activity and antitumor effect of pegylated recombinant human interleukin 2 (PEG-rhIL-2) obtained by site-specific conjugation of polyethylene glycol (PEG) with non-natural amino acids, and to explore its antitumor mechanism. METHODS: The binding activities of PEG-rhIL-2 at three different sites (T41, Y45, and V91) to human interleukin 2 receptors α (IL-2R_α) and β (IL-2R_β) and were detected by surface plasmon resonance (SPR) technology. Western blot was used to detect the levels of the Janus kinase-signal transducer and activator of transcription 5 (JAK-STAT5) signaling pathway activated by different doses of rhIL-2 and PEG-rhIL-2 in CTTL-2 and YT cells. Blood was collected after a single administration in mice to detect the drug concentration at different time points and evaluate the pharmacokinetic parameters of Y45-PEG-rhIL-2. Mouse hepatoma cell line Hepa1-6, pancreatic cancer cell line Pan-02, and colon cancer cell line MC-38 were selected. Tumor models were constructed in C57BL/6 mice. Different doses of Y45-PEG-rhIL-2 and excipient control were administrated respectively to evaluate the tumor suppression effect of the drug. In the MC-38 colon cancer model, the tumor suppression effect of Y45-PEG-rhIL-2 combined with anti-programmed death-1 (PD-1) monoclonal antibody was evaluated. Hepa1-6 mouse tumor models were constructed and rhIL-2, Y45-rhIL-2 and Y45-PEG-rhIL-2 were administrated respectively. The proportion of tumor-infiltrating lymphocytes was analyzed by flow cytometry. RESULTS: The SPR detection results showed that the binding activities of PEG-rhIL-2 to IL-2R_α/IL-2R_β were both reduced. The affinity of Y45-PEG-rhIL-2 to IL-2R_α was reduced to approximately 1/250, and its affinity to IL-2R_β was reduced to 1/3. Western blot results showed that the activity of Y45-PEG-rhIL-2 in stimulating JAK-STAT5 signaling in CTLL-2 cells expressing heterotrimeric IL-2 receptor complex IL-2R_αβγwas reduced to approximately 1/300, while its activity in YT cells expressing heterodimeric IL-2 receptor complex IL-2R_βγwas reduced to approximately 1/3. The pharmacokinetic evaluation after a single dose in the mice showed that the elimination half-life of Y45-PEG-rhIL-2 was 17.7 h. Y45-PEG-rhIL-2 has pharmacokinetic characteristics superior to those of rhIL-2. Y45-PEG-rhIL-2 showed dose-dependent tumor suppression activity, and the combination of Y45-PEG-rhIL-2 and anti-PD-1 antibody had a better tumor-inhibiting effect than the single use of Y45-PEG-rhIL-2 or anti-PD-1 antibody. Flow cytometry analysis demonstrated that 72 h after the administration of Y45-PEG-rhIL-2, the proportion of tumor-infiltrating cytotoxic T lymphocytes (CD8⁺T cells) increased by 86.84%. At 120 h after administration, the ratio of CD8⁺T cells to regulatory T cells (Treg) increased by 75.10%. CONCLUSION Y45-PEG-rhIL-2 obtained by site-specific conjugation via non-natural amino acids changed its receptor binding activity and inhibited tumor growth in dose-dependent manner in multiple tumor models by regulating CD8⁺T cells.
Interleukin-2/pharmacokinetics* ; Animals ; Mice ; Humans ; Recombinant Proteins/pharmacology* ; Polyethylene Glycols/chemistry* ; Cell Line, Tumor ; Antineoplastic Agents/pharmacokinetics* ; Signal Transduction/drug effects* ; STAT5 Transcription Factor/metabolism* ; Interleukin-2 Receptor alpha Subunit/metabolism* ; Interleukin-2 Receptor beta Subunit/metabolism*

Dihydromyricetin(DMY),a flavonoid compound extracted from Ampelopsis grossedentata,exhibits diverse pharmacological activities,including anti-inflammatory,antioxidant,antitumor,neuroprotective,and immuno-modulatory effects.Recent studies have demonstrated that DMY can suppress inflammatory responses and oxidative stress through multiple molecular pathways,as well as regulate bile acid metabolism and maintain intestinal microbiota balance.These actions help reduce histopathological damage,improve gastrointestinal barrier function,and alleviate the symptoms of digestive system diseases.DMY has shown significant therapeutic effects in the treatment of gastrointestinal inflamma-tion,liver diseases,and digestive tract tumors.This review systematically summarizes the mechanisms of action and thera-peutic potential of DMY in digestive system diseases,providing a scientific basis and theoretical support for its clinical ap-plication and the development of new drugs.

Urinary incontinence is the most common complication after radical prostatectomy.Traditional Chinese medicine treatments for this disease,such as acupuncture,are diverse and have definite therapeutic effects.The disease is located in the bladder,and its etiology and pathogenesis are mostly related to the dysfunction of the kidney,spleen,triple energizer and other viscera.In view of the insufficiency of kidney essence,decline of the gate of vitality fire(mingmen fire),Qi deficiency leading to impaired bladder containment,compounded by obstruction of meridians from pathogenic factors including blood stasis,damp-heat,and phlegm turbidity.The"Shuanggu Yitong"acupuncture achieves"Dual fortification"through tonifying the kidney and primordial qi consolidation,combined with spleen strengthening and qi supplementation.Additionally,"purging"targets the elimination of blood stasis,phlegm turbidity,damp-heat and other solid evils.This integrated approach combines acupuncture and moxibustion modalities with reinforcing-reducing needle manipulation techniques,simultaneously addressing both the symptoms and root causes of the problem.By coordinating pathogen elimination with constitutional regulation,this method demonstrates favorable therapeutic outcomes in clinical practice and is worthy of further clinical promotion.

Objective To investigate the key targets and pathways of Juhongtai formula granules in improving acute lung injury(ALI).Methods Juhongtai formula granules and their drug-containing serum components were analyzed by UPLC-QTOF-MS/MS,and the active ingredients were screened based on the"Lipinski Rule of Five".The action targets of the above active ingredients were obtained through the TCMSP,Swiss Target Prediction and SuperPred databases,and the ALI-related targets were obtained from the GeneCards,OMIM,PharmGKB,CTD databases and GEO chip.The target protein-protein interaction network was constructed using the String database and Cytoscape to analyze the key targets.The DAVID database was used for GO functional annotation and KEGG pathway enrichment analysis,and the CB-dock2 platform was used for molecular docking verification.After one week of adaptive feeding,the experimental SD rats were randomly divided into the blank group,the model group,the positive control group,the negative control group and the Juhongtai formula granules group,with 6 rats in each group.The rat model of ALI was replicated by tracheal infusion of lipopolysaccharide.The pathological morphology of lung tissues in each group of rats was observed by HE staining,and the mRNA expressions of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1 β(IL-1β)and nitric oxide were detected by reverse transcription quantitative polymerase chain reaction(RT-qPCR).Results In total,44 components of the Juhongtai formula granules were identified,including 26 drug-containing serum components,and 21 key compounds were screened.A total of 22 intersection targets were obtained.The pathway enrichment results indicated the action of these targets on the advanced glycation end product receptor(AGE-RAGE)and TNF signaling pathways.The docking results showed that limonin,kaempferol and naringenin could be matched with prostaglandin peroxidase 2,nitric oxide synthase 2 and TNF.Animal experiments confirmed that the Juhongtai formula granules can alleviate inflammatory infiltration in lung tissue and reduce the expressions of TNF-α,IL6,IL-1β mRNA and nitric oxide.Conclusion Juhongtai formula granules can inhibit ALI-related inflammatory targets through the synergistic effects of multiple components such as limonin and kaempferol,regulate signaling pathways such as AGE-RAGE and TNF,reduce the production of inflammatory factors and nitric oxide,and improve ALI.

Objective To investigate the key targets and pathways of Juhongtai formula granules in improving acute lung injury(ALI).Methods Juhongtai formula granules and their drug-containing serum components were analyzed by UPLC-QTOF-MS/MS,and the active ingredients were screened based on the"Lipinski Rule of Five".The action targets of the above active ingredients were obtained through the TCMSP,Swiss Target Prediction and SuperPred databases,and the ALI-related targets were obtained from the GeneCards,OMIM,PharmGKB,CTD databases and GEO chip.The target protein-protein interaction network was constructed using the String database and Cytoscape to analyze the key targets.The DAVID database was used for GO functional annotation and KEGG pathway enrichment analysis,and the CB-dock2 platform was used for molecular docking verification.After one week of adaptive feeding,the experimental SD rats were randomly divided into the blank group,the model group,the positive control group,the negative control group and the Juhongtai formula granules group,with 6 rats in each group.The rat model of ALI was replicated by tracheal infusion of lipopolysaccharide.The pathological morphology of lung tissues in each group of rats was observed by HE staining,and the mRNA expressions of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1 β(IL-1β)and nitric oxide were detected by reverse transcription quantitative polymerase chain reaction(RT-qPCR).Results In total,44 components of the Juhongtai formula granules were identified,including 26 drug-containing serum components,and 21 key compounds were screened.A total of 22 intersection targets were obtained.The pathway enrichment results indicated the action of these targets on the advanced glycation end product receptor(AGE-RAGE)and TNF signaling pathways.The docking results showed that limonin,kaempferol and naringenin could be matched with prostaglandin peroxidase 2,nitric oxide synthase 2 and TNF.Animal experiments confirmed that the Juhongtai formula granules can alleviate inflammatory infiltration in lung tissue and reduce the expressions of TNF-α,IL6,IL-1β mRNA and nitric oxide.Conclusion Juhongtai formula granules can inhibit ALI-related inflammatory targets through the synergistic effects of multiple components such as limonin and kaempferol,regulate signaling pathways such as AGE-RAGE and TNF,reduce the production of inflammatory factors and nitric oxide,and improve ALI.

Dihydromyricetin(DMY),a flavonoid compound extracted from Ampelopsis grossedentata,exhibits diverse pharmacological activities,including anti-inflammatory,antioxidant,antitumor,neuroprotective,and immuno-modulatory effects.Recent studies have demonstrated that DMY can suppress inflammatory responses and oxidative stress through multiple molecular pathways,as well as regulate bile acid metabolism and maintain intestinal microbiota balance.These actions help reduce histopathological damage,improve gastrointestinal barrier function,and alleviate the symptoms of digestive system diseases.DMY has shown significant therapeutic effects in the treatment of gastrointestinal inflamma-tion,liver diseases,and digestive tract tumors.This review systematically summarizes the mechanisms of action and thera-peutic potential of DMY in digestive system diseases,providing a scientific basis and theoretical support for its clinical ap-plication and the development of new drugs.

Urinary incontinence is the most common complication after radical prostatectomy.Traditional Chinese medicine treatments for this disease,such as acupuncture,are diverse and have definite therapeutic effects.The disease is located in the bladder,and its etiology and pathogenesis are mostly related to the dysfunction of the kidney,spleen,triple energizer and other viscera.In view of the insufficiency of kidney essence,decline of the gate of vitality fire(mingmen fire),Qi deficiency leading to impaired bladder containment,compounded by obstruction of meridians from pathogenic factors including blood stasis,damp-heat,and phlegm turbidity.The"Shuanggu Yitong"acupuncture achieves"Dual fortification"through tonifying the kidney and primordial qi consolidation,combined with spleen strengthening and qi supplementation.Additionally,"purging"targets the elimination of blood stasis,phlegm turbidity,damp-heat and other solid evils.This integrated approach combines acupuncture and moxibustion modalities with reinforcing-reducing needle manipulation techniques,simultaneously addressing both the symptoms and root causes of the problem.By coordinating pathogen elimination with constitutional regulation,this method demonstrates favorable therapeutic outcomes in clinical practice and is worthy of further clinical promotion.

Objective:To analyze the clinical characteristics of nail involvement in patients with pemphigus and the correlation between nail damage and the severity of pemphigus.Methods:The clinical data of 23 patients with pemphigus combined with nail damage admitted to the People′s Hospital of Xinjiang Uygur Autonomous Region from January 2011 to August 2019 were retrospectively analyzed, the manifestations of pemphigus combined with nail damage were summarized. The nail damage number of patients with pemphigus complicated with nail damage of different genders were analyzed, as well as the distribution of nail damage in nail and toenail. The titer of anti-desmocoglycoprotein(dsg) antibody was detected in all patients. The relationship between nail damage and disease severity and course in pemphigus patients was analyzed.Results:A total of 132 damage nails were found in 23 patients (14 males and 9 females) with pemphigus, including 66 nails in male, 66 nails in female, 82 nails and 50 toenails. There were 10 forms of nail damage, of which chronic paronychia was the most common. The number of damage nails between different genders in pemphigus patients was statistically significant (χ 2=9.183, P<0.001). The distribution of nail and toenail damage in pemphigus patients was statistically significant (χ 2=10.880, P<0.001). Of the 23 patients, only 3 were positive for dsg1 and 1 was positive for dsg3. There were 19 patients with both positive for dsg1 and dsg3. The titers of dsg1 and dsg3 were compared in 14 patients before and after nail damage. The results showed that the titers of anti-dsg antibody in pemphigus patients after nail damage were significantly higher than before. Thirteen of the 23 patients had nail damage at the time of the initial onset of pemphigus. The nail damage occurred from 6 weeks before the onset to 4 weeks after the onset. The nail damage occurred in 10 patients when the disease recurred. The nail damage occurred within 4 weeks before or at the same time with the blister. Conclusions:The number of damage nails per capita in female patients with pemphigus and nail damage was significantly higher than that in male patients, and nail damage was more common. The titers of anti-dsg antibody will be at a high level when pemphigus patients with nail damage, and the condition gets worse. Nail involvement is positive to the severity of the disease, and it can prolong the time of disease.

【Objective】 To summarize a rapid and effective method to eliminate autoantibodies interference with blood group identification and evaluate its treatment effect. 【Methods】 Blood samples with suspicious results in initial blood group identification were collected, and those caused by autoantibodies were chosen, and their red blood cells were washed, dispersed or treated with sulfhydryl reagent. After allogeneic or autologous absorption of plasma, blood groups of those patients were re-detected to evaluate the effectiveness of the above method. 【Results】 Among 39 patients presenting suspicious ABO blood group, 9 were interfered by autoantibodies. After appropriate treatment, the ABO/RhD blood group of those patients could be identified. 【Conclusion】 Autoantibodies could interfere the identification of ABO/RhD blood group, and the efficiency and accuracy of blood group identification could be improved by analyzing the test results and selecting appropriate treatment methods.

Cell-Free Nucleic Acids ; blood ; Female ; Humans ; Mosaicism ; Placenta ; Pregnancy