MRGPRX2 antagonists possess the potential for the treatment of allergic rhinitis, atopic dermatitis, and chronic urticaria. Previously, we identified a class of diaryl urea (DPU) MRGPRX2 antagonists with sub-micromolar IC₅₀ values in vitro. However, the structure-activity relationship remains unclear. Herein, we adopted a "relative symmetry with electronegativity of different key-groups" strategy for further modification of DPUs to achieve a promising MRGPRX2 antagonist with higher activity and safety. Electrostatic potential energy analysis and biological evaluation revealed that B-1023 and B-5023, that possess relatively symmetric electron-withdrawing substituents, remarkable inhibited mast cell degranulation at a sub-micromolar IC₅₀ in vitro and alleviated anaphylactic symptoms. Furthermore, B-1023, mitigated antigen-induced pulmonary inflammation (AIPI) in mice and competitively bonded to MRGPRX2. In summary, the "relative symmetry with electronegativity of different key-groups" strategy provided a drug design pattern for MRGPRX2 antagonists and identified promising antiallergic precursors for AIPI treatment.

OBJECTIVES: To explore the value of ferroptose-related genes in the diagnosis and prediction of ulcerative colitis (UC). METHODS: We used UC dataset from the GEO database to screen for differentially expressed genes (DEGs) in UC. The DEGs related to ferroptositis were screened from the FerrDb database and their functions were analyzed. The hub genes were identified by constructing the protein-protein interaction network (PPI), the differences in immune infiltration levels between UC and the control group were evaluated using CIBERSORT, and the diagnostic values of the hub genes for UC were verified by using the training set. In a mouse model of UC, we examined the expression levels of the hub genes in the colon tissues of the mice using real-time fluorescence quantitative PCR (qPCR). RESULTS: We identified a total of 76 DEGs related to ferroptosis. Functional enrichment analysis showed that these genes were significantly enriched in ferroptosis and hypoxia pathways. The PPI network identified 10 hub genes, and 9 of them were highly expressed in UC. Analysis of immune cell infiltration showed that 27 cell types were significantly increased in UC (P<0.05), and the immune checkpoints-related genes had the strongest correlation with the hub gene PPARG (P<0.05). Verification analysis using the training set showed that P4HB, PPARG and STAT3 had the best predictive value for UC (P<0.05). In the UC mouse model, the expression of PPARG was significantly decreased and the expressions of P4HB and STAT3 were significantly increased in the colon tissues of the mice as compared with the normal mice. CONCLUSIONS Ferroptose-related genes have significant value for diagnosis and prediction of UC.
Colitis, Ulcerative/genetics* ; Animals ; Mice ; Ferroptosis/genetics* ; Humans ; Protein Interaction Maps ; Disease Models, Animal ; Gene Expression Profiling ; STAT3 Transcription Factor/genetics*

Objective:To explore the deubiquitination modification of the deubiquitinase BRCC3 in rats with neuropathic pain (NPP) treated with meridian tuina of Zhuang medicine and its analgesic molecular mechanism.Methods:Rats were divided into normal group, sham-operation group, model group, sham-tuina group, and meridian tuina of Zhuang medicine group using a random number table method, with 9 rats in each group. Except for the normal and sham operation groups, spinal nerve ligation models were prepared in all other groups. On the first day after surgery, intervention was carried out on the meridian tuina of Zhuang medicine and sham-tuina groups for 14 days, while the other three groups were not intervened; the paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) of each group of rats were measured at 0 days before operation, 1, 7, and 14 days after operation. Western blot was used to detect the expressions of BRCC3 and NLRP3 proteins in spinal cord tissue, and ELISA was used to detect the level of IL-1β in serum.Results:On postoperative 7 and 14 d, compared with the model group, the meridian tuina of Zhuang medicine group showed an increase in PWMT and PWTL, a decrease in NLPR3 and BRCC3 expression in spinal cord tissue, and a decrease in serum IL-1β levels ( P<0.05). Compared with the sham-tuina group, the meridian tuina of Zhuang medicine group showed an increase in PWMT and PWTL, a decrease in NLRP3 protein expression in spinal cord tissue, and a decrease in IL-1β levels in serum ( P<0.05). Conclusion:Meridian tuina of Zhuang medicine can alleviate pain sensitivity in SNL model rats, and its mechanism is related to the inhibition of the expression of deubiquitinase BRCC3 by meridian massage of Zhuang medicine, which increases the ubiquitination level of NLRP3 and hinders its activation, thereby blocking the immune inflammatory response mediated by inflammatory factors.

Objective To investigate the expression of Tropomyosin 4(TPM4)in thyroid cancer and its effects on the invasion and migration of thyroid cancer cells.Methods The expression level and prognostic value of TPM4 in thyroid cancer were analyzed based on bioinformatics,and its functional involvement was explored through Gene Set Enrichment Analysis(GSEA).In thyroid cancer K1 cells,lentiviral transfection was performed to establish the experimental group(TPM4 shRNA),the negative control group(empty lentiviral transfection),and the control group(untreated).Cell viability and proliferation were assessed using CCK-8 and BrdU assays.Transwell migration and invasion assays were performed to evaluate the effects of TPM4 on the migratory and invasive capacities of thyroid cancer cells.Results TPM4 expression was significantly upregulated in thyroid cancer(P<0.05)and correlated with TNM staging(P<0.05).Patients with higher TPM4 expression in advanced TNM stages exhibited poorer prognosis(P<0.05).GSEA results indicated that high TPM4 expression was enriched in gene sets associated with epithelial-mesenchymal transition,inflammatory response,P53 signaling pathway,and cell cycle.Following TPM4 knockdown in K1 cells,thyroid cancer cell growth was slowed(P<0.01),proliferative activity was decreased(P<0.001),and invasion and migration abilities were significantly impaired(P<0.001).Conclusion TPM4 is highly expressed in thyroid cancer and promotes the invasion and migration capabilities of thyroid cancer cells.

Objective To investigate the acute toxicology and intervention effect of Panacis Majoris Rhizoma on rats with chronic pharyngitis.Methods A single,maximum dose of Panacis Majoris Rhizoma(74.4 g·kg-1)was administered to Kunming mice to evaluate its toxicity,involving the assessment of the survival status of the mice,organ indices,morphological changes in major organs,blood routine,and biochemical indicators.SD rats were randomly divided into the control group,model group,prednisone group(6.25 mg·kg-1),and low-,medium-,and high-dose Panacis Majoris Rhizoma groups(0.58,1.16,and 2.32 g·kg-1).All rats received the corresponding drugs(or normal saline)via intragastric administration once daily for a duration of 30 days.Except the control group,chronic pharyngitis was induced in rats of the other groups by using β-hemolytic streptococcus.Following euthanasia,serum inflammatory levels of interleukin-6(IL-6),cyclooxygenase-2(COX-2),interleukin-1β(IL-1β),intercellular adhesion molecule-1(ICAM-1),C-reactive protein(CRP),tumor necrosis factor(TNF-α),monocyte chemoattractant protein-1(MCP-1),and prostaglandin E2(PGE2)were measured.Additionally,pharyngeal tissues were stained with HE and pathological characteristics were observed.Results Toxicological studies have demonstrated that the administration of Panacis Majoris Rhizoma resulted in significant increase in plasma alanine transaminase levels and spleen index of mice,along with corresponding tissue pathological alterations.Nevertheless,no noteworthy pathological changes were observed in other organs,and there were no notable changes in blood routine and plasma biochemical indicators.Pharmacodynamic investigations have revealed that Panacis Maioris Rhizoma effectively reduces the serum levels of inflammatory factors and improves pathological changes in pharyngeal tissues.Conclusion Panacis Maioris Rhizoma alleviated β-hemolytic streptococcus-induced CP by inhibiting inflammatory responses,and may show potential toxicity to the spleen.

Objective:To carry out preimplantation genetic testing (PGT) for a Chinese pedigree affected with Rett syndrome (RTT).Methods:A pedigree affected with RTT who had presented at the First Hospital of Jilin University on June 4, 2021 was selected as the study subject. Variant of the MECP2 gene was analyzed by next generation sequencing (NGS) and Sanger sequencing. Direct sequencing was also used to determine the carrier status for the c. 925C>T variant of the MECP2 gene in the blastocysts, and Sanger sequencing was used to validate the results. The MECP2 gene and 168 effective single nucleotide polymorphism (SNP) loci within 2 Mb ranges up- and downstream of the gene were used to construct a haplotype for analyzing the variant site in the embryos, and embryos without the variant were subjected to the analysis for chromosomal aneuploidies. Results:PGT analysis revealed that five out of seven blastocysts did not harbor the pathogenic variant. The results of aneuploidy analysis indicated that two out of five blastocysts without the variant were euploid. Following genetic counselling, the couple had opted to transplant the optimal blastocyst. Following clinical pregnancy, prenatal diagnosis showed that the fetus has a normal chromosomal karyotype, and the c. 925C>T variant was not detected in the amniotic fluid sample. A healthy girl was born by Cesarean section at full term.Conclusion:NGS can attain efficient PGT detection and reduce the risk of disease recurrence in families affected with RTT.

Objective To explore the protective effect of Lycium barbanun glycopeptide(LbGP)against osteogenic inhibition induced by exosomes derived from human gingival fibroblasts(HGFs)exposed to radiation.Methods Cultured HGFs with or without LbGP pretreatment were exposed to 8 Gy X-ray radiation,and the changes in cell apoptosis,senescence and α-SMA level were detected using RT-qPCR,Western blotting and β-galactosidase staining.The exosomes secreted by the treated cells were extracted,and after identification by electron microscopy,particle size analysis and Western blotting,the exosomes were added into primary cultured bone mesenchymal stem cells(BMSCs),and osteoclast activity and osteogenesis in the cell cultures were detected by Trap staining,Alizarin red staining,ALP staining,RT-qPCR and Western blotting.Results In cultured HGFs,X-ray radiation significantly increased the percentage of senescent cells,which was obviously lowered by LbGP treatment.X-ray radiation significantly reduced Bcl-2/Bax ratio and increased α-SMA expression in HGFs,and these changes were significantly suppressed by LbGP pretreatment.In rat BMSCs,incubation with the exosomes derived from HGFs with radiation exposure caused a significant increase of osteoclasts,reduced calcium nodules and lowered alkaline phosphatase expression in the cells;The opposite changes were observed in the cells treated with exosomes from LbGP-pretreated HGFs,which also significantly increased the cellular expressions of the osteogenic genes(BMP2,ALP,and RUNX2)and proteins(ALP and RUNX2)as compared with the exosomes from irradiated HGFs.Conclusion LbGP can effectively inhibit osteoclast activity and promote osteogenesis by acting on exosomes secreted by irradiated HGFs,suggesting its potential value for treatment of osteoradionecrosis of the jaw.

Primary prostate signet ring cell carcinoma (PPSRCC) is one of the extremely rare malignant tumors in the male urogenital system, and its diagnosis mainly relies on pathological and immunohistochemical examination. Compared with typical prostate cancer, PPSRCC is characterized by more aggressive with less treatment response and poor prognosis. Current researches on the pathogenesis, clinical manifestations, pathological characteristics, diagnosis, treatment and prognosis of PPSRCC were reviewed.

ObjectiveBased on the network pharmacology system and quantitative spectroscopy of traditional Chinese medicine（TCM） compounds， a topological network analysis method with equilibrium constant as the core was established to further explore the interaction between allergenic components and their network targets in Shuanghuanglian injection（SHLI）， in order to provide new ideas and experimental basis for identifying and screening potential allergens of SHLI. MethodAfter one week of adaptive feeding， 72 SPF-grade SD male rats were randomly divided into blank group， SHLI standard group， Lonicerae Japonicae Flos（LJF） group， Scutellariae Radix（SR） group， Forsythiae Fructus（FF） group， and 7 groups of SHLI matching groups（groups 1-7）， with 6 rats in each group. Rats in each group were administered the drug intravenously and blood samples were taken after steady state， high performance liquid chromatography（HPLC） characterization profiles of the testing drugs and plasma components in each group were established， and the peak area changes of the drugs and plasma components in each group were calculated after the component groups were classified. Enzyme-linked immunosorbent assay（ELISA） was used to determine the changes of immunoglobulin E（IgE）， histamine（HIS）， tryptase（TPS）， total complement（CH50） and terminal complement complex（C5b-9） in animal blood samples. MATLAB R2020b v9.9.0 software was used to calculate the network balance constants of the component groups with the targets， and the eigenvalues of the matrices composed of network equilibrium constants were calculated and ranked according to their values. ResultELISA results showed that， compared with the blank group， groups 1-3 could significantly increase the IgE level， groups 1-2， groups 4-6 and SHLI standard group could significantly increase the HIS level， group 4 could significantly increase the CH50 level， groups 1， 3-4， LJF group and FF group could significantly increase the TPS level， SR group could significantly increase the C5b-9 level， and the differences were all statistically significant（P<0.05）. According to the retention time of chromatographic peaks， it was classified into 6 component groups from C1 to C6 by HPLC. The order of the network balance constants of each component group was C6>C4>C1>C5>C3>C2， indicating that C6 had the greatest effect on the allergic reaction， and was most likely to be the allergen. The sequence of eigenvalues was C2>C5b-9>C3>C1>CH50>C6>C5>IgE>TPS>C4>HIS， indicating that component group C2 had the greatest contribution to the whole network. ConclusionBased on the correlation analysis of SHLI component group and allergy-like target network， this study clarified that component group C6 may be a potential allergen in SHLI， and the component group C2 may be a key node in the mechanism of drug action， which can provide new strategies and methods for the screening of allergens in TCM injections.

Objective To explore the protective effect of Lycium barbanun glycopeptide(LbGP)against osteogenic inhibition induced by exosomes derived from human gingival fibroblasts(HGFs)exposed to radiation.Methods Cultured HGFs with or without LbGP pretreatment were exposed to 8 Gy X-ray radiation,and the changes in cell apoptosis,senescence and α-SMA level were detected using RT-qPCR,Western blotting and β-galactosidase staining.The exosomes secreted by the treated cells were extracted,and after identification by electron microscopy,particle size analysis and Western blotting,the exosomes were added into primary cultured bone mesenchymal stem cells(BMSCs),and osteoclast activity and osteogenesis in the cell cultures were detected by Trap staining,Alizarin red staining,ALP staining,RT-qPCR and Western blotting.Results In cultured HGFs,X-ray radiation significantly increased the percentage of senescent cells,which was obviously lowered by LbGP treatment.X-ray radiation significantly reduced Bcl-2/Bax ratio and increased α-SMA expression in HGFs,and these changes were significantly suppressed by LbGP pretreatment.In rat BMSCs,incubation with the exosomes derived from HGFs with radiation exposure caused a significant increase of osteoclasts,reduced calcium nodules and lowered alkaline phosphatase expression in the cells;The opposite changes were observed in the cells treated with exosomes from LbGP-pretreated HGFs,which also significantly increased the cellular expressions of the osteogenic genes(BMP2,ALP,and RUNX2)and proteins(ALP and RUNX2)as compared with the exosomes from irradiated HGFs.Conclusion LbGP can effectively inhibit osteoclast activity and promote osteogenesis by acting on exosomes secreted by irradiated HGFs,suggesting its potential value for treatment of osteoradionecrosis of the jaw.