Background Existing studies have confirmed that fine particulate matter (PM_2.5)is one of the important factors inducing pulmonary fibrosis. Pulmonary fibrosis is the terminal stage of a major category of lung diseases characterized by the destruction of tissue structure, and eventually leading lung ventilation and ventilation dysfunction. No effective pulmonary fibrosis treatment is available yet. Objective To investigate the protective effect of salidroside on pulmonary fibrosis induced by the exposure of PM_2.5 and its molecular mechanism. Methods Seventy 7-week-old male C57BL/6 mice were randomly divided into four groups: control group (intratracheal instillation of normal saline + saline by gavage, n=25), Sal group (intratracheal instillation of normal saline + Sal 60 mg·kg⁻¹ by gavage, n=10), PM_2.5 group (intratracheal instillation of PM_2.5 5 mg·kg⁻¹ + saline by gavage, n=10), and Sal + PM_2.5 group (intratracheal instillation of PM_2.5 5 mg·kg⁻¹ +Sal 60 mg·kg⁻¹ by gavage, n=10). The mice were administered by gavage once daily, intratracheal instillation once every 3 d, and every 3 d constituted an experimental cycle. At the end of the 26-30th cycles, 3 mice in the control group and 3 mice in the PM_2.5 group were randomly sacrificed, and the lung tissues were collected for Masson staining to verify whether the pulmonary fibrosis model was successfully established. After 30 cycles, the model was successfully constructed. After 1 week of continuous observation, the mice were sacrificed, and the blood and lung tissues of the mice were collected to make lung tissue sections. Assay kits were correspondingly employed to detect oxidative stress indicators such as serum malondialdehyde (MDA) and superoxide dismutase (SOD). Western blotting was used to detect the expression of fibrosis-related proteins (Collagen-III, α-SMA), mitochondrial dynamics-related proteins (MFN1, Drp1), and mitophagy-related proteins (PINK1, Parkin, and LC3). Results Compared with the control group, the weight gain rate of the PM_2.5 group was slowed down (P<0.05), which was alleviated by the Sal intervention (P<0.05). The lung coefficient increased after the PM_2.5 exposure (P<0.05), which was alleviated by Sal intervention. Compared with the control group, the PM_2.5 group showed severe alveolar structure damage, inflammatory cell infiltration, and blue collagen deposition, and significantly increased the lung injury score, collagen volume fraction (CVF), Szapiel score, and Ashcroft score (P<0.05), as well as serum oxidative stress levels (P<0.05). The protein expression levels of Collagen-III, α-SMA, Drp1, PINK1, Parkin, and LC3 II/I were increased (P<0.05), and the expression of MFN1 was decreased (P<0.05). Compared with the PM_2.5 group, the Sal intervention alleviated lung injury, reduced inflammatory cell infiltration and collagen deposition, showing decreased lung injury score, CVF, Szapiel score, and Ashcroft score (P<0.05), and decreased serum oxidative stress levels (P<0.05); the protein expression levels of Collagen-III, α-SMA, PINK1, Parkin, and LC3 II/I were decreased (P<0.05), the expression level of Drp1 was decreased, and the expression level of MFN1 was increased. Conclusion In the process of pulmonary fibrosis induced by PM_2.5 exposure in mice, Sal may affect mitochondrial autophagy through PINK1/Parkin pathway and play a protective role. The specific mechanism needs to be further verified.

As a common medical condition， vertigo can be induced by multiple diseases in the modern medical system. Its incidence rate shows an upward trend with the increase in age. According to the theory of mutual interference between clear Qi and turbid Qi in Huangdi's Internal Classic （Huang Di Nei Jing）， this paper systematically analyzes the pathogenesis of vertigo and explores the mechanism and clinical application value of Xiao Chaihutang in the treatment of vertigo. It is believed that the mutual inference between clear Qi and turbid Qi leads to the failure of clear Yang to ascend， resulting in the lack of nourishment for the brain and the inability of turbid Yin to descend， which disturbs the clear orifices， thus causing vertigo. The core pathogenesis lies in the dysfunction of Qi movement， the disorder of body fluid distribution， and the imbalance between Yin and Yang. The compatibility of Xiao Chaihutang takes into account the methods of pungent medicinal materials opening and bitter medicinal materials descending， tonifying deficiency and purging excess， and regulating Qi movement. This prescription can regulate the pathological state of the mutual interference between clear Qi and turbid Qi from three aspects： regulating Qi movement throughout the body， harmonizing the distribution of body fluids， and coordinating Yin and Yang as well as the interior and exterior， thus preventing and treating vertigo. Modern research findings show that Xiao Chaihutang can improve hemodynamics to promote cerebral blood circulation and has anti-inflammation， immunomodulatory， and anti-tumor functions， which correspond to the therapeutic effects of Xiao Chaihutang under the theory of mutual interference between clear Qi and turbid Qi. The decoction exerts therapeutic effects on vertigo caused by hypertension， stroke， otitis media， Meniere’s disease， and brain tumor as well as benign paroxysmal positional vertigo. Further exploration of the theoretical connotation of mutual inference between clear Qi and turbid Qi and analysis of the pathogenesis of vertigo and the therapeutic effect of Xiao Chaihutang can better interpret the internal correlations among the three， thus providing new ideas for the syndrome differentiation and treatment of vertigo.

Objective:To evaluate the role of the SIRT1/FoxO1 signaling pathway in trilobatin-induced reduction of cerebral ischemia-reperfusion (I/R) injury in rats.Methods:Eighty clean-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 230-280 g, were divided into 4 groups ( n=20 each) using a random number table method: sham operation group (group S), cerebral I/R group (group CIR), trilobatin+ cerebral I/R group (group T) and trilobatin+ cerebral I/R+ SIRT1/FoxO1 signaling pathway inhibitor EX527 group (group E). The model of focal cerebral I/R injury was established by middle cerebral artery occlusion in anesthetized animals. Trilobatin 15 mg/kg was given by gavage twice a day for 3 consecutive days starting from 3 days before ischemia in T and E groups. EX527 5 mg/kg was intraperitoneally injected before each gavage in group E. Modified Longa scoring scale was used to assess neurological function at 24 h of reperfusion, then the rats were sacrificed and whole brain tissues were obtained for determination of cerebral infarct size (using TTC staining), apoptosis rate and level of reactive oxygen species (ROS) in the hippocampus (by flow cytometry), expression of SIRT1 and acetylated FOXO1 (Ac-FOXO1) (by Western blot) and contents of superoxide dismutase (SOD) and malondialdehyde (MDA) (by enzyme-linked immunosorbent assay) and for microscopic examination of pathological changes in the hippocampal CAI area after HE staining. Results:Compared with group S, Longa score, cerebral infarct size, apoptosis rate of hippocampal neurons, and levels of ROS and MDA were significantly increased, the content of SOD was decreased, the expression of SIRT1 was down-regulated, and the expression of Ac-FOXO1 was up-regulated in group CIR ( P<0.05). Compared with group CIR, Longa score, cerebral infarct size, apoptosis rate of hippocampal neurons, and levels of ROS and MDA were significantly decreased, the content of SOD was increased, the expression of SIRT1 was up-regulated, and the expression of Ac-FOXO1 was down-regulated in group T ( P<0.05). Compared with group T, Longa score, cerebral infarct size, apoptosis rate of hippocampal neurons, and levels of ROS and MDA were significantly increased, the content of SOD was decreased, the expression of SIRT1 was down-regulated, and the expression of Ac-FOXO1 was up-regulated in group E ( P<0.05). Conclusions:Trilobatin may inhibit oxidative stress responses and neuronal apoptosis in hippocampi by activating the SIRT1/FoxO1 signaling pathway, thus alleviating cerebral I/R injury in rats.

Objective:To explore the mechanism of Yupingfeng Powder on patients with chronic obstructive pulmonary disease (COPD) through UHPLC-QE-MS combined with network pharmacology and molecular docking technology.Methods:UHPLC-QE-MS technology was used to detect the chemical composition of Yupingfeng Powder, and its targets were obtained using ChEMB and TCMIO databases; COPD treatment targets were obtained from TTD, GeneCards, SymMap, and DisGeNET databases, and the intersection of the two was taken. GO enrichment analysis and KEGG pathway analysis were performed on intersection targets. Top 20 metabolites with the most matching targets were selected, their corresponding targets with the top 20 KEGG pathway targets were merged, and potential targets for the treatment of COPD with Yupingfeng Powder were obtained. A target protein-target protein interaction network (PPI) was constructed, key targets of Yupingfeng Powder for the treatment of COPD were screened, and molecular docking was used for validation.Results:Under positive and negative ion modes, a total of 73 active components were identified in Yupingfeng Powder. The top three ranked in terms of degree were genistein, apigenin, and kaempferol. 449 targets of 73 active components and 3 455 disease targets were obtained from the database. The intersection of the two was obtained, resulting in 294 common targets. A total of 69 pathways were identified through KEGG pathway analysis, involved in the pathogenesis of cancer, non-small cell lung cancer signaling pathway, VEGF signaling pathway, T cell receptor (TCR) signaling pathway, arachidonic acid metabolism, Toll like receptor signaling pathway, etc. GO enrichment analysis obtained 2 647 biological processes, 126 cellular components, and 289 molecular functions, involving reactions to nutritional levels, extracellular stimuli, oxidative stress, etc. The top 20 KEGG pathways and the top 20 metabolites with the most matching targets correspond to a total of 281 targets. The key targets were TP53, STAT3, c-Jun, AKT1, and ESR1. The molecular docking results showed that genistein, apigenin, and kaempferol bound well with core targets TP53, STAT3, and c-Jun.Conclusion:Yupingfeng Powder may affect the levels of inflammation, immune regulation, and oxidative stress in COPD through multiple components, targets, and pathways, thereby affecting the progression of COPD.

Long non-coding RNAs (lncRNAs) reportedly function as important modulators of gene regulation and malignant processes in the development of human cancers. The lncRNA JPX is a novel molecular switch for X chromosome inactivation and differentially expressed JPX has exhibited certain clinical correlations in several cancers. Notably, JPX participates in cancer growth, metastasis, and chemoresistance, by acting as a competing endogenous RNA for microRNA, interacting with proteins, and regulating some specific signaling pathways. Moreover, JPX may serve as a potential biomarker and therapeutic target for the diagnosis, prognosis, and treatment of cancer. The present article summarizes our current understanding of the structure, expression, and function of JPX in malignant cancer processes and discusses its molecular mechanisms and potential applications in cancer biology and medicine.
Humans ; RNA, Long Noncoding/genetics* ; Neoplasms/genetics* ; MicroRNAs/genetics* ; Gene Expression Regulation ; X Chromosome Inactivation

In surgery of patients with hypertensive intracerebral hemorrhage, precise positioning and minimally invasive operation provide a strong guarantee for overall curative effect. As an emerging visualization software, 3D-Slicer can optimize surgical approach, achieve precise intraoperative positioning, and accurately measure hematoma volume to guide treatment plan implementation. This article reviews the application of 3D-Slicer in surgery of patients with hypertensive intracerebral hemorrhage.

OBJECTIVE: To explore the genetic etiology and clinical outcome of a child with steroid-resistant nephrotic syndrome and diffuse mesangial sclerosis. METHODS: Genomic DNA was extracted from peripheral blood leukocytes of the proband and his parents. Targeted capture - next generation sequencing and Sanger sequencing were carried out. Candidate variant was verified by segregation analysis in his family. RESULTS: A heterozygous missense variant of the TRPC6 gene, namely c.325G>A (p.Gly109Ser), was detected in the proband. The same variant was not detected in either parent. According to the guidelines for the interpretation of sequence variants developed by American College of Medical Genetics and Genomics, the variant was predicted as pathogenic. CONCLUSION The missense variant of the TRPC6 gene probably underlay the diffuse mesangial sclerosis in this patient. Above finding has expanded the phenotypic spectrum of the TRPC6 gene.
Child ; Genomics ; Humans ; Mutation, Missense ; Nephrotic Syndrome/genetics* ; Sclerosis ; TRPC6 Cation Channel/genetics*

Objective:To investigate the molecular pathogenesis of a newly discovered gene mutation in a family with hereditary coagulation factor Ⅺ(FⅪ) deficiency.Methods:The proband was admitted to the First Affiliated Hospital of Wenzhou Medical University in September 2021 due to "calculus of intrahepatic duct". The patient had no symptoms of spontaneous bleeding.The clinical data and blood samples of the proband and her family members (10 persons in 3 generations) were collected.The activated partial thromboplastin time (APTT) and FⅪ activity (FⅪ:C) were performed by the one-stage clotting assay. FⅪ antigen (FⅪ:Ag) were detected by enzyme linked immunosorbent assay (ELISA). Genomic DNA extracted from peripheral blood cells of subjects was used as template to analyze F11 gene mutation by DNA direct sequencing. Bioinformatics software was used to analyze the effects of mutations on protein structure and function. Wild-type and mutant FⅪ protein expression vectors were constructed and transient transfected into HEK293T cells. The total RNA was extracted from positive transfected cells and then reversely transcribed into cDNA. The mRNA expression level of F11 gene in transfected cells was detected by real-time fluorescence quantitative PCR (qRT-PCR). The content of FⅪ:Ag and the expression of FⅪ protein in transfected cell lysates and culture supernatant were detected by ELISA and western blot.Results:The APTT of the proband was significantly prolonged to 107.9s (reference range 29.0-43.0s), while FⅪ:C and FⅪ:Ag were significantly decreased to 2% (reference range 84%-122%) and 5% (reference range 76%-127%), respectively. Gene sequencing analysis indicated that the proband had c.536C>T (p.Thr161Met) heterozygous missense mutation and c.1556G>A (p.Trp501Ter) heterozygous nonsense mutation in exon 6 and 13 of the F11 gene, respectively. Bioinformatics analysis showed that the amino acids at site 161 of FⅪ protein were threonine (Thr) in the matrix composed of five different species, indicating that Thr161 site was highly conserved among homologous genes in different species. p.Thr161Met heterozygous mutation affected the stability of local intermolecular structure of FⅪ protein. In vitro expression experiments of p.Thr161Met mutation showed that FⅪ protein had a normal synthesis in the cells but secretion dysfunction.Conclusions:c.536C>T (p.Thr161Met) heterozygous missense mutation and c.1556G>A (p.Trp501Ter) heterozygous nonsense mutation were mainly responsible for the decrease of FⅪ in this family. p.Thr161Met mutation was first reported in the world and did not affect the normal synthesis of FⅪ protein, but caused secretion dysfunction.

Objective:To explore the molecular pathogenesis of hereditary protein C (PC) deficiency due to a p. Gly86Asp variant of the PROC gene through in vitro expression experiment.Methods:Wild type and Gly86Asp mutant expression plasmids of PC were constructed and respectively transfected into HEK 293FT cells. Total RNA was extracted from the transfected cells, and the expression of PROC gene was determined by quantitative real-time PCR (qRT-PCR). PC antigen (PC: Ag) in the supernatant of cell culture and cell lysate was determined by enzyme-linked immunosorbent assay (ELISA), and the level of PC protein was detected by Western blotting. Results:qRT-PCR has detected no significant difference in the transcription level of wild-type and mutant-type PC. Compared with the wild type, the level of mutant PC: Ag in the supernatant and cell lysate were 81.3%±2.6% and 110.0%±2.8%, respectively. No difference was detected in the molecular weight between the wild-type and mutant-type PC by Western blotting. The PC content of mutant type was higher than wild-type in cell lysate, while the opposite was found with the cell culture supernatant.Conclusion:The impaired secretion by mutant PC may be the molecular mechanism of PC deficiency caused by the p.

OBJECTIVE: To explore the molecular basis for a Chinese pedigree affected with hereditary coagulation factor VII (FVII) deficiency. METHODS: The coding regions of F7 gene were amplified by PCR and sequenced. Suspected variants were confirmed by reverse sequencing and validated in other members from the pedigree. Pathogenicity of the variants was analyzed with multiple bioinformatic tools. RESULTS: Genetic analysis revealed that the proband has carried compound heterozygous c.985T>C (p.Ser329Pro) and c.1091G>A (p.Arg364Gln) variants in exon 8 of the F7 gene. Her mother, brother and son were heterozygous for c.985T>C (p.Ser329Pro), while her father was heterozygous for c.1091G>A (p.Arg364Gln). Phylogenetic analysis suggested that both p.Ser329 and p.Arg364 are highly conserved among homologous species. Online bioinformatic software predicted both variants to be deleterious. Protein model analysis suggested that the Pro329 side chain may form a new hydrogen bond with Leu333. The Pro benzene ring may clash with Glu325 in the p.Ser329Pro variant model. The p.Arg364Gln variant have two additional hydrogen bonds compared with wild type Arg364. Both variants may lead to alteration of the protein structure. CONCLUSION The p.Ser329Pro and p.Arg364Gln variants in exon 8 of the F7 gene probably account for the reduced FVII in this pedigree.