Objective To observe the influence of exosomal miR-155 derived from monocytes stimulated by heat stress on the inflammatory response of hepatocytes. Methods According to different treatments, we termed THP-1 monocytes into the control group, the heat stress group, and the ulinasatin group. We then extracted the exosomes from each group. To study the function of exsome on hepatocytes, we incubated hepatocytes with these exosomes. We then tested the alanine aminotransferase and lactate dehydrogenase levels in the supernatant, measured cell viability, and detected the relative mRNA levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the hepatocytes. Then, the expression changes of exosomal miR-155 derived from monocytes were detected using RT-qPCR. Furthermore, the potential target of miR-155 was analyzed using database retrieval, dual-luciferase report, and Western blotting assay. Finally, the effect of monocyte exosomes on hepatocyte miR-155 and the treatment with ulinastatin or miR-155 inhibitor on hepatocyte injury were observed. Results When incubating hepatocytes with the exosomes derived from monocytes stimulated by heat stress, the levels of alanine aminotransferase and lactate dehydrogenase in the supernatant and the mRNA levels of TNF-α and IL-6 in hepatocytes were all significantly increased (P<0.01) with decreased cell survival rate (P<0.05). Heat stress increased the level of miR-155 contained in monocytes and their release of exosomes. Database retrieval, dual-luciferase reports, and Western blotting assay showed that miR-155 might target SOCS1. The exosomes from monocytes stimulated by heat stress increased miR-155 in liver cells. Ulinastatin or miR-155 inhibitor could reduce the exosomal miR-155 level and decrease TNF-α and IL-6 mRNA expression in liver cells. The difference was statistically significant (P<0.05). Conclusion The monocytes stimulated by heat stress increase the inflammatory response of hepatocytes, which may be associated with increased exosomal miR-155 using SOCS1 as a potential target. Ulinastatin may relieve the hepatocytic mRNA expression of inflammatory cytokines in this setting.

Objective To analyze the drug resistance features of 118 patients with bone and joint tuberculosis. Methods The clinical data of 118 joint tuberculosis patients who were hospitalized in Beijing Chest Hospital from January 2016 to January 2022 were retrospectively analyzed. Drug susceptibility test was performed for the following 16 drugs streptomycin (Sm), isoniazid (INH), rifampicin (RFP), ethambutol (EMB), rifapentine (Rft), levofloxacin (Lfx), amikacin (Am), capreomycin (Cm), prothionamide (Pto), isoniazid aminosalicylate (Pa), moxifloxacin (Mfx), p-aminosalicylic acid (PAS), clarithromycin (Clr), rifabutin (Rfb), kanamycin (Km) and clofazimine (Cfz). Analyze the Mycobacterium tuberculosis culture results, drug sensitivity results, initial or retreatment status of patients with bone and joint tuberculosis, as well as the drug resistance types of patients diagnosed with bone and joint tuberculosis by etiology. Results The total drug resistance rate of 118 bone and joint tuberculosis patients to at least one of the 16 drugs was 28.0%(33/118), of which the drug resistance rate was significantly higher in previously treated patients than in new patients with statistically significant difference [70.0%(21/30) vs. 13.6%(12/88), P<0.001]. The top seven drug resistance rate of bone and joint tuberculosis to the 16 drugs were: Sm, INH, RFP, Rft, Rfb, Pa and Clr, and the top seven drug resistance rates of new patients were: Sm, INH, Clr, Pa, RFP, Rft and PAS, and the top seven drug resistance rates of in previously treated patients were: Sm, RFP, Rft, Rfb, INH, Pa and EMB. The mono-resistance rate of bone and joint tuberculosis, poly-drug resistance rate of spinal tuberculosis, and multidrug-resistance rate were 7.6%(9/118), 8.5%(10/118), and 11.9%(14/118), respectively. There was no significant difference of the mono-resistance rate of bone and joint tuberculosis between new patients and in previously treated patients [6.8%(6/88) vs. 10.0%(3/30), P=0.691], but the poly-drug resistance rate and the multidrug-resistance rate were significantly higher in previously treated patients than in new patients, and the differences were statistically significant [20.0%(6/30) vs. 4.5%(4/88), P=0.017; 40.0%(12/30) vs. 2.3%(2/88), P<0.001]. Conclusions There was serious epidemic of drug resistance in bone and joint tuberculosis. While performing surgical treatment, clinicians should develop effective drug treatment regimens according to the results of drug sensitivity tests.

Sepsis is a serious disease with high incidence and mortality. The pathogenesis, clinical manifestations, and multiple organ dysfunction of sepsis are complex and diverse, and this high heterogeneity leads to many challenges in the treatment of sepsis. The ideal treatment of sepsis should be based on its subphenotypes and targeted to the specific one. This review summarizes the research progress on phenotyping of adult sepsis from the relevant definitions, research methods, existing subtypes (such as based on molecular mechanism, pathophysiological mechanism, clinical manifestations, etc.), and the possible new subtypes in the future. We expect to flash a light on the discovery of new subtypes and the basic clinical research for the treatment of septic subtypes through this paper.

Objective To analyze the relationship between small intestinal bacterial overgrowth (SIBO) and gastroesophageal reflux disease (GERD). Method A total of 5832 patients who visited the Department of Gastroenterology, the Sixth Medical Center of Chinese PLA General Hospital from August 2019 to August 2021 were selected. The patients were divided into GERD group (n=1752) and non-GERD group (n=4080) according to gastroesophageal reflux disease. The two groups were compared for general features and SIBO prevalence. Subgroup analysis was performed in GERD group. The gastroesophageal reflux disease questionnaire (GerdQ) scores between the SIBO-positive group (n=1051) and the SIBO-negative group (n=701) were compared. The prevalence of SIBO was compared between the group with proton pump inhibitor (PPI) (n=1280) and the group without PPI (n=472). The prevalence of SIBO was compared between patients in non-erosive esophagitis (n=1051), erosive esophagitis (n=643) and Barrett's esophagus (n=58). Risk factors for GERD were analyzed by multivariate logistic regression. Results Age, body mass index, GerdQ score, smoking and prevalence of SIBO in GERD group were higher than those in non-GERD group (P<0.05). Multivariate analysis found that SIBO, obesity, drinking and smoking were risk factors for GERD. Subgroup analysis showed that the GerdQ score in SIBO-positive group (9.54±1.59) was higher than that in SIBO-negative group (8.40±1.54, P<0.05). The prevalence of SIBO in patients taking PPI (64.9%) was higher than that in patients without PPI (46.6%, P<0.05); The prevalence of SIBO in patients with erosive esophagitis (68.7%) and Barrett's esophagus (69.0%) was higher than that in patients with non-erosive esophagitis (54.1%, P<0.05). Conclusions SIBO is risk factor for GERD. Reflux symptoms are more severe when GERD patients have SIBO.

Objective To screen, identify and validate the immunoreactive epitopes on the glycoprotein-N terminal (Gn) of Hantaan virus (HTNV) for providing a new idea for prevention of hemorrhagic fever with renal syndrome (HFRS). Methods The Gn protein sequences of HTNV strain 76-118 were obtained from UniProt database. IEDB, SMMPMBEC, NetMHCpan 4.1, SYFPEITHI and Rankpep were used to predict epitope affinity. Immunogenicity was analyzed by VaxiJen. Blastp analyzed the conservation; HPEPDOCK and EpiDOCK simulated pMHC docking; TBtools implemented bidirectional cluster analysis; Immunoreactivity of epitope in vivo was evaluated by enzyme-linked immunospot assay. Results The Gn protein sequence of HTNV 76-118 strain (PRO_0000036816) was obtained from UniProt database. Five affinity algorithms were integrated to obtain 61 dominant epitopes in mouse H-2 subtype, 234 dominant epitopes in human major histocompatibility complex (MHC)-Ⅰ subtype, and 212 dominant epitopes in MHC-Ⅱ subtype. VaxiJen screening obtained 23, 110 and 42 dominant epitopes of H-2, MHC-Ⅰ and MHC-Ⅱ subtypes, respectively. Further Blastp screening resulted in 3 MHC-Ⅰ restricted epitopes with high affinity and strong immunogenicity, and 81 MHC-Ⅱ restricted epitopes conserved between species. Bidirectional hierarchical cluster analysis revealed the similarity of HTNV Gn epitopes in H2-d, H2-b of mice and some human leucocyte antigen (HLA). ELISpot verified that 5 epitopes could induce splenic cells to secrete interferon-gamma (IFN-γ). Conclusions The present study predicted and verified the cellular immunoreactive epitopes on HTNV Gn that can induce cellular immune reactivity, revealed the cross activity across-genes, species and genera immunoreactivity of viral antigens in MHC presentation, and provide guidance for the development of novel HFRS epitope vaccines.

Objective To investigate the prognostic value of C-C motif chemokine ligand 11 (CCL11) and midkine (MK) in serum of patients with differentiated thyroid carcinoma (DTC). Methods One hundred and fifty patients with DTC admitted in Henan Provincial People's Hospital from January 2015 to January 2017 were selected as DTC group, 150 patients with benign thyroid disease in the same period were selected as benign group, and 150 healthy volunteers were selected as control group. The serum CCL11 and MK levels of the three groups were compared. Patients in DTC group were divided into survival subgroup and death subgroup according to their prognosis. The clinical data between survival and death patients were compared. Cox regression analysis was used to analyze the factors affecting the prognosis of DTC patients. The receiver operating characteristic (ROC) curve was established to evaluate the diagnostic value of serum CCL11 and MK levels in the prognosis of DTC. Results Compared with control group, the levels of serum CCL11 and MK increased in DTC group and benign group, and the levels were higher in DTC group than in benign group (P<0.05). On the 3rd day after operation, the levels of serum CCL11 and MK were lower in both DTC group and benign group than those at the time of diagnosis, and still higher in DTC group than in benign group (P<0.05). In DTC group, 135 patients survived and 15 died. The age, tumor diameter, TNM stage, lymph node metastasis, capsular invasion, differentiation degree and serum thyroglobulin (Tg), CCL11, MK levels were significantly different (P<0.05) between the survived and dead patients with DTC. Cox regression analysis showed that TNM stage, lymph node metastasis and serum Tg, CCL11, MK levels were the prognostic factors of DTC (P<0.05). The results of ROC analysis showed that serum CCL11 and MK levels were of high value in diagnosis of DTC prognosis, and the diagnostic efficiency was higher when they were combine used (sensitivity 93.33%, specificity 73.33%, AUC 0.835). Conclusions The serum levels of CCL11 and MK were abnormally elevated in patients with DTC. The combined detection of serum levels of CCL11 and MK might have higher prognostic diagnostic value for DTC.

Objective The study aimed to investigate the association between the genetic variant of neurofibromatosis 2 (NF2) gene and susceptibility to meningioma in Chinese population and analyze the correlation between NF2 gene polymorphism and meningioma. Methods 215 patients who had underwent neurosurgical treatment and been histologically diagnosed as meningioma were included as the case group in Department of Neurosurgery, Beijing Tiantan Hospital affiliated to Capital Medical University between May 2010 to January 2011. The NF2 gene polymorphisms including rs2530673 and rs2530662 were analyzed by Multiplex SNaPshot methods. The correlation between NF2 gene polymorphism and prognosis of meningioma was analyzed. Results The NF2 gene polymorphisms rs2530673 and rs2530662 are not significant associated with the susceptibility to meningioma (P>0.05). According to the WHO grade I pathological subtypes, the result of stratification analysis showed that the patients carrying NF2 gene polymorphism loci rs2530673 CC genotype reduced the risk of transitional meningiomas in the dominant model distribution (OR=0.506, 95%CI 0.266-0.962, P=0.036). Besides, no significant association was found between NF2 gene polymorphism loci rs2530673 and rs2530662 and the recurrence of meningioma (P>0.05). For tumor location, the result of stratification analysis showed that the rs2530673 polymorphism of NF2 gene was associated with postoperative recurrence of meningiomas and patients carrying the rs2530673 CC genotype had shorter progression free survival time and higher recurrence risk in non-skull base meningiomas (all P<0.05). No significant correlation was found between rs2530673/rs2530662 polymorphisms and the prognosis of meningioma (P>0.05). Conclusions The C allele of rs2530673 may be a protective factor for the oneset of transitional meningioma. There may be no significant association between NF2 gene rs2530673/rs2530662 polymorphisms and the susceptibility in meningioma. However, the rs2530673 polymorphism of NF2 gene was associated with postoperative recurrence of meningiomas and patients carrying the rs2530673 CC genotype had higher recurrence risk in non-skull base meningiomas.

Objective To investigate the molecular mechanism of histone demethylase 5C (KDM5C) regulating human cervical carcinogenesis through Hippo-YAP1 pathway. Methods Using CaSki cell lines stably overexpressing KDM5C protein, whole transcriptome sequencing was performed by RNA-Seq technique, and differentially expressed genes were analyzed, and then GO analysis, KEGG analysis and protein interaction network analysis were performed on these differential genes. After that, siRNA knocked down KDM5C and reversely verified the expression changes of key regulated gene Yes associate protein 1 (YAP1) by RT-qPCR and Western blotting. Meanwhile, in CaSki cell lines, the effect of KDM5C protein overexpression on the methylation status of YAP1 gene promoter region was analyzed by chromatin immunoprecipitation sequencing (ChIP-Seq) and ChIP-qPCR methods. Results RNA-Seq analysis showed that overexpression of KDM5C significantly up-regulated expressions of 356 mRNAs and down-regulated 335 mRNAs expressions (P<0.05). GO enrichment analysis showed that KDM5C protein was mainly involved in various biological development processes of the body. KEGG enrichment analysis showed that KDM5C protein was mainly involved in focal adhesion, steroid hormone biosynthesis, Hippo-YAP1 pathway, FoxO pathway, apoptosis and infection. Further RT-qPCR analysis showed that knockdown of KDM5C with gene specific siRNAs could up-regulate the expression of YAP1, and Western blotting results also confirmed that reduction the expression of KDM5C protein could up-regulate the levels of YAP1 and phosphorylated YAP1 simultaneously (P<0.05). ChIP-Seq analysis showed that KDM5C overexpression cell line could significantly increase the H3K4me1 level and decrease the H3K4me3 level in the promoter interval of YAP1 gene compared with the control cell line, and this expression change was also verified by subsequent ChIP-qPCR (P<0.05). Conclusions The KDM5C protein regulates the methylation level of the histone H3K4me1/me3 in the YAP1 gene promoter of cervical cancer cells, thereby affecting the transcription of the YAP1 gene. As a core factor in Hippo-YAP1 pathway, the expression of YAP1 protein directly affects cell adhesion, proliferation, and apoptosis, thereby participating in the occurrence and development of cervical cancer.

Objective To investigate the effect of lycopene (Lyc) on inflammatory factors and intestinal mucosal barrier function in mice with traumatic brain injury (TBI). Methods Forty-five SPF level of adult male BALB/c mice were randomly divided into sham group, TBI group and TBI+Lyc group, with 15 mice in each group. The TBI model was prepared by modified compression injury model, and the mice in sham group underwent an identical process without mechanical trauma. Each group was further divided into 5 subgroups at 1 d, 2 d, 3 d, 7 d and 14 d after modelling (n=3/subgroup). Mice in sham group and TBI group were treated with 10 mg/kg sunflower oil, and in TBI+Lyc group were treated with 10 mg/kg sunflower oil-dissolved Lyc (concentration 1 mg/ml), daily gavage at a fixed time for 14 d. The inferior vena cava blood and ileum tissue of mice in each subgroup were collected. The serum levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), intestinal fatty acid binding protein (I-FABP) and diamine oxidase (DAO) were detected by ELISA, and the serum levels of D-lactic acid (D-LA) and endotoxin were detected by colorimetric method and endpoint chromogenic assay, respectively. Pearson correlation test was used to analyze the correlation between inflammatory factors and intestinal mucosal barrier function indicators. The pathological changes of ileum tissues were observed by HE staining, and the villus height and crypt depth of ileum were measured. Results The serum levels of TNF-α, IL-1β, I-FABP, D-LA, DAO and endotoxin of mice in TBI group and TBI+Lyc group gradually decreased with time, but were obviously higher than those in sham group at different time points (P<0.01 or P<0.001); the serum levels of TNF-α, IL-1β, I-FABP, D-LA, DAO and endotoxin of mice in TBI+Lyc group were significantly lower than those in TBI group at different time points (P<0.01 or P<0.001). Serum TNF-α and IL-1β were significantly positively correlated with I-FABP, D-LA, DAO and endotoxin, respectively (P<0.01). The sham group showed basically normal ileal villi, the structure of ileal villi in TBI group was obviously damaged, and the structure of ileal villi in TBI+Lyc group was basically neat. The ileal villus height and crypt depth of mice in TBI group were lower than those in sham group at each time points (P<0.05), and the ileal villus height and crypt depth of mice in TBI+Lyc group were significantly higher than those in TBI group at different time points (P<0.05). Conclusion Lyc can reduce the levels of inflammatory factors in various stages of TBI mice and may improve the intestinal mucosal barrier function by inhibiting inflammatory response.

Objective To explore the effect of versican (VCAN) gene on immunocyte infiltration and prognosis of bladder urothelial carcinoma (BUC). Methods The high-throughput sequencing data and clinical data of BUC were downloaded from TCGA database. The correlation between the expression level of VCAN and clinical characteristics of BUC patients was analyzed using the Limma R package; univariate and multivariate Cox regression analysis were performed to analyze the effect of VCAN expression and clinical characteristics (including gender, age, tumor grade, tumor stage, lymph node metastasis and distant metastasis) on the prognosis of BUC patients; TIMER database was used to analyze the correlation between VCAN expression level and immunocyte infiltration. T24 cells were divided into three groups: control group (cells transfected without any siRNA), NC siRNA group (cells transfected with negative control siRNA), and VCAN siRNA group (cells transfected with VCAN siRNA). After transfection, the proliferation, invasion and cell cycle proportion of T24 cells were determined by MTT assay, Transwell assay and flow cytometry, respectively. The protein expression levels of Cyclin E, Cyclin D1, E-cadherin and matrix metalloproteinase-9 (MMP-9) were determined by Western blotting. Results Compared with normal bladder tissues, the expression level of VCAN mRNA in BUC tissues was significantly up-regulated, and was positively correlated with tumor grade, tumor stage and distant metastasis (P<0.01). High VCAN expression was an independent risk factor for poor prognosis in BUC patients (P<0.05). The results of TIMER database analysis showed that the expression level of VCAN was significantly positively correlated with the infiltration degree of macrophages and regulatory T cells in BUC (P<0.001). Compared with control group, the proliferation and invasion ability of T24 cells in VCAN siRNA group decreased significantly and the cell proportion of G₀/G₁ phase increased significantly (P<0.05), the protein expression levels of Cyclin E, Cyclin D1 and MMP-9 were down-regulated significantly, and of E-cadherin was up-regulated significantly (P<0.05). Conclusions VCAN is highly expressed in BUC and correlated with immune infiltration and prognosis of BUC. Silencing the expression of VCAN may significantly inhibit the proliferation and invasion of T24 cells by regulating the protein expression levels of Cyclin E, Cyclin D1, E-cadherin and MMP-9.