Analysis of intestinal flora in elderly Uygur patients with sarcopenia.

OBJECTIVE: This study was designed to analyze the structural characteristics of the intestinal flora of elderly Uygur patients with sarcopenia, thereby providing new ideas for clinical treatment. METHODS: Firstly, fecal samples were collected from 40 elderly Uygur patients with sarcopenia (Sarcopenia group) and 40 healthy people (Control group). Next, significant differences in the intestinal flora between the two groups were analyzed based on 16S rDNA high-throughput sequencing. The linear discriminant analysis effect size (LEfSe) was used to estimate the magnitude of the effect of each component (species) abundance on the differential effect. Additionally, an analysis was also performed on the relationship between the intestinal flora and the cytokines in the peripheral blood of patients with sarcopenia. RESULTS: The results of ß diversity showed that there were differences in the structure of the intestinal flora between the two groups. Besides, the phylum level of intestinal flora between the two groups was not significantly different. However, the difference was significant in the intestinal flora at the order, family, and genus levels between the two groups. Among them, Lachnoclostridium, Photobacterium, Anaerobic Bacillus, Hydrogenophilus, and Eubacterium were enriched in the Sarcopenia group; Prevotella 9, Firmicutes FCS020 group, Streptobacillus, Aggregatibacter, Corynebacterium, Clostridium Difficile, and Haloanaerobium were enriched in the Control group. The LEfSe outcomes further showed that Lachnoclostridium was highly enriched in the Sarcopenia group; Prevotella 9 and Firmicutes FCS020 group were significantly enriched in the Control group. Furthermore, the relative abundance of Lachnoclostridium and Streptobacillus were significantly different in patients with high and low IL-6 levels. CONCLUSION: In conclusion, Lachnoclostridium is significantly enriched in the intestines of elderly Uygur patients with sarcopenia; the increase in Lachnoclostridium abundance and the decrease in Streptobacillus abundance are associated with high levels of IL-6. Therefore, abnormal intestinal flora is related to inflammatory reflexes in patients with sarcopenia.

Prediction of survival and immunotherapy response by the combined classifier of G protein-coupled receptors and tumor microenvironment in melanoma.

BACKGROUND: Melanoma is the deadliest form of skin tumor, and G protein-coupled receptors (GPCRs) play crucial roles in its carcinogenesis. Furthermore, the tumor microenvironment (TME) affects the overall survival (OS) and the response to immunotherapy. The combination of GPCRs and TME from a multi-omics perspective may help to predict the survival of the melanoma patients and their response to immunotherapy. METHODS: Bulk-seq, single-cell RNA sequencing (scRNA-seq), gene mutations, immunotherapy responses, and clinicopathologic feature data were downloaded from public databases, and prognostic GPCRs and immune cells were screened using multiple machine learning algorithms. The expression levels of GPCRs were detected using real-time quantitative polymerase chain reaction (qPCR) in A375 and HaCaT cell lines. The GPCR-TME classifier was constructed and verified using different cohorts and multi-omics. Gene set enrichment analysis (GSEA), weighted gene co-expression network analysis (WGCNA), and tracking tumor immunophenotype (TIP) were used to identify the key biological pathways among the GPCR-TME subgroups. Then, tumor mutational burden (TMB), vital mutant genes, antigen presentation genes, and immune checkpoints were compared among the subgroups. Finally, the differences in immunotherapy response rates among the GPCR-TME subgroups were investigated. RESULTS: A total of 12 GPCRs and five immune cell types were screened to establish the GPCR-TME classifier. No significant differences in the expression levels of the 12 GPCRs were found in the two cell lines. Patients with high GPCR score or low TME score had a poor OS; thus, the GPCRlow/TMEhigh subgroup had the most favorable OS. The scRNA-seq result revealed that immune cells had a higher GPCR score than tumor and stromal cells. The GPCR-TME classifier acted as an independent prognostic factor for melanoma. GSEA, WGCNA, and TIP demonstrated that the GPCRlow/TMEhigh subgroup was related to the activation and recruitment of anti-tumor immune cells and the positive regulation of the immune response. From a genomic perspective, the GPCRlow/TMEhigh subgroup had higher TMB, and different mutant genes. Ultimately, higher expression levels of antigen presentation genes and immune checkpoints were observed in the GPCRlow/TMEhigh subgroup, and the melanoma immunotherapy cohorts confirmed that the response rate was highest in the GPCRlow/TMEhigh cohort. CONCLUSIONS: We have developed a GPCR-TME classifier that could predict the OS and immunotherapy response of patients with melanoma highly effectively based on multi-omics analysis.

Fluvastatin combined with corbrin capsule can improve depression and anxiety in the treatment of chronic obstructive pulmonary disease.

OBJECTIVE: To investigate the effect of fluvastatin (Flu) combined with corbrin capsule (CC) on the pulmonary function (PF) in patients with chronic obstructive pulmonary disease (COPD). METHODS: Totally, 156 patients with COPD treated in our hospital were assigned: 86 patients in the research group (RG), who were treated with Flu plus CC, and 70 patients in the control group (CG), who were treated with CC plus conventional drugs. The changes in inflammatory factors, including interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α) and procalcitonin (PCT), of the two groups before and after treatment were compared. The complications, psychological status, quality of life (QOL) and recurrence rate of the two groups were analyzed. RESULTS: The total effective rate in the RG was dramatically higher than that in the CG (P<0.05). Compared with the factors in the CG, the PF in the RG notably increased after treatment (P<0.05); the blood gas levels were noticeably better (P<0.05); and the level of inflammatory factors decreased (P<0.05). The incidence of complications in the RG was noticeably lower than that in the CG (P<0.05). The psychological status and QOL in the RG were remarkably better than those in the CG (P<0.05), and the recurrence rate within one year of diagnosis was lower than that in the CG (P<0.05). CONCLUSION: Flu combined with CC is effective and safe in the treatment of COPD and can effectively improve the PF and the QOL of patients.

MUC1 downregulation promotes TNF-α-induced necroptosis in human bronchial epithelial cells via regulation of the RIPK1/RIPK3 pathway.

MUC1 (mucin 1), a membrane-tethered mucin glycoprotein, is highly expressed on the surface of respiratory epithelial cells and plays a key role in anti-inflammatory and antiapoptotic responses against infections. However, little is known about the link between MUC1 and necroptosis in asthma. This study aimed to investigate the effects of MUC1 on TNF-α-induced necroptosis in human bronchial epithelial (16HBE) cells and the underlying molecular mechanism. Negative control and MUC1-siRNA cells were treated with TNF-α in the presence or absence of necrostatin-1 (Nec-1). Necroptosis was investigated using flow cytometry analyses, and the protein expression levels of MUC1, receptor-interacting protein kinase-1 (RIPK1), RIPK3, and phosphorylated RIPK1 were detected by western blot analysis. In addition, the interactions between RIPK and MUC1 were analyzed by coimmunoprecipitation. The results demonstrated that TNF-α could induce necroptosis of 16HBE cells, and MUC1 expression was increased upon treatment with TNF-α. The coimmunoprecipitation outcomes showed that MUC1 interacted with RIPK1 but not with RIPK3 in 16HBE cells, and the interaction was augmented by TNF-α. Furthermore, MUC1 downregulation obviously increased the TNF-α-induced necroptosis of 16HBE cells and enhanced the expression of p-RIPK1-Ser166 and RIPK3, whereas these phenomena were partially attenuated by Nec-1. These results may provide a new insight into the mechanism of severe asthma-related necroptosis and lay a foundation for the future development of new anti-inflammatory drugs for asthma.

Cyclic peptide *CRRETAWAC* attenuates fibronectin-induced cytokine secretion of human airway smooth muscle cells by inhibiting FAK and p38 MAPK.

α5ß1 integrin is highly expressed in airway smooth muscle cells and mediate the adhesion of airway smooth muscle cells to fibronectin to regulate airway remodelling in asthma. This study aimed to investigate the effects of synthetic cyclic peptide *CRRETAWAC* on fibronectin-induced cytokine secretion of airway smooth muscle cells and the underlying mechanism. Human airway smooth muscle cells were isolated and treated with fibronectin, IL-13, *CRRETAWAC* peptide, α5ß1 integrin-blocking antibody, FAK inhibitor or p38 MAPK inhibitor. The transcription and secretion of eotaxin-1 and RANTES were detected by real-time PCR and ELISA, respectively. The phosphorylation of FAK and MAPKs including p38, ERK1/2 and JNK1/2 was detected by Western blot analysis. The transcription and secretion of eotaxin-1 and RANTES increased in airway smooth muscle cells cultured in fibronectin-coated plates. However, α5ß1 integrin-blocking antibody, *CRRETAWAC* peptide, FAK inhibitor or p38 MAPK inhibitor significantly reduced mRNA levels and the secretion of eotaxin-1 and RANTES in airway smooth muscle cells cultured in fibronectin-coated plates. In addition, the phosphorylation of FAK and p38 MAPK was significantly increased in airway smooth muscle cells cultured in fibronectin-coated plates compared to the cells cultured in uncoated plates and was significantly reduced in airway smooth muscle cells treated with *CRRETAWAC* peptide. Fibronectin induces cytokine synthesis and secretion of airway smooth muscle cells. Peptide *CRRETAWAC* antagonizes fibronectin-induced cytokine synthesis and secretion of airway smooth muscle cells via the inhibition of FAK and p38 MAPK, and is a potential agent for the therapy of asthma.

Correlation between TCRCalpha -560 C/T polymorphism and the clinical presentation of Uygur IgA nephropathy patients in XinJiang.

OBJECTIVE: To investigate the relationship between T cell receptor alpha chain constant gene (TCRCalpha) -560 C/T polymorphism and the clinical presentation of Uygur IgA Nephropathy patients in XinJiang. METHODS: TCRCalpha -560 C/T genotypes were determined by PCR-RFLP in 300 Chinese Uygur IgAN patients and 600 healthy Chinese Uygur control subjects. All subjects were classified, based on their genotype, into TT, CT and CC groups and their corresponding clinical presentation was analyzed. RESULTS: No significant difference was observed in the frequency of CC/CT/TT genotypes in patients and control subjects (chi2 = 0.904, P = 0.636). However, the incidence of intermittent microscopic hematuria and proteinuria is significantly higher in patients with CT genotype than CC and TT genotypes (chi2 = 33.978, P < 0.05). CONCLUSION: TCRCalpha-560 C/T gene polymerphism may be associated with the occurrence of intermittent microscopic hematuria and proteinuria in Chinese Uygur IgAN patients.