Objective:To investigate the effects of long-term exposure to chrysotile and crocidolite on miRNAs and epithelial mesenchymal transformation (EMT) -related gene expression in human pleural mesothelial cells.Methods:In November 2020, fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of EMT-related genes in human pleural mesothelioma cells (NCl-H2052 cells, NCl-H2452 cells) and human normal mesothelial cells (Met-5A cells). MiRNAs with abnormal expression in human pleural mesothelioma cells were screened out from the previous miRNA chip data of research group, and target genes of differentially expressed miRNAs were predicted using miRWalk database (http: //mirwalk.umm.uni-heidelberg.de). RT-qPCR was used to verify the abnormal expression of EMT-related miRNAs in cell lines. Met-5A cells were treated with 5μg/cm 2 chrysotile and crocidolite respectively for 48 h a time, once a week and a total of 10 times. Chrysotile group, crocidolite group and control group were set up. And the control group was added with the same volume of PBS. The expression changes of EMT-related genes and abnormal expression miRNAs in each group were detected by RT-qPCR. The differences among the groups were compared by one-way ANOVA, and the differences between the control group and the experimental group were compared by dunnet- t test. Results:Compared with Met-5A cells, the expression levels of Vimentin and Twist genes were increased, and the expression level of E-cadherin genes was decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001). Target genes of miRNAs with abnormal expression in miRNA chip were predicted, and the results showed four abnormally expressed miRNAs associated with EMT and verified the expression of these four miRNAs in the cell lines. Compared with Met-5A cells, the expression level of hsa-miR-155-5p was increased in NCl-H2052 cells and NCl-H2452 cells, the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001), which was consistent with the results of chip analysis. After exposure of Met-5A cells, it was found that compared with the control group, the expression levels of Vimentin and Twist genes, hsa-miR-155-5p, hsa-miR-34b-5p and hsa-miR-34c-5p in the crocidolite group were increased, while the expression level of E-cadherin gene was decreased ( P<0.05). Compared with the control group, the expression levels of Vimentin, Twist and E-cadherin genes in chrysotile group were increased, while the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased ( P<0.05) . Conclusion:Long-term exposure to chrysotile and crocidolite could cause Met-5A cells to produce miRNAs and EMT-related gene expression changes similar to mesothelioma cells.

Objective:To investigate the effects of long-term exposure to chrysotile and crocidolite on miRNAs and epithelial mesenchymal transformation (EMT) -related gene expression in human pleural mesothelial cells.Methods:In November 2020, fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of EMT-related genes in human pleural mesothelioma cells (NCl-H2052 cells, NCl-H2452 cells) and human normal mesothelial cells (Met-5A cells). MiRNAs with abnormal expression in human pleural mesothelioma cells were screened out from the previous miRNA chip data of research group, and target genes of differentially expressed miRNAs were predicted using miRWalk database (http: //mirwalk.umm.uni-heidelberg.de). RT-qPCR was used to verify the abnormal expression of EMT-related miRNAs in cell lines. Met-5A cells were treated with 5μg/cm 2 chrysotile and crocidolite respectively for 48 h a time, once a week and a total of 10 times. Chrysotile group, crocidolite group and control group were set up. And the control group was added with the same volume of PBS. The expression changes of EMT-related genes and abnormal expression miRNAs in each group were detected by RT-qPCR. The differences among the groups were compared by one-way ANOVA, and the differences between the control group and the experimental group were compared by dunnet- t test. Results:Compared with Met-5A cells, the expression levels of Vimentin and Twist genes were increased, and the expression level of E-cadherin genes was decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001). Target genes of miRNAs with abnormal expression in miRNA chip were predicted, and the results showed four abnormally expressed miRNAs associated with EMT and verified the expression of these four miRNAs in the cell lines. Compared with Met-5A cells, the expression level of hsa-miR-155-5p was increased in NCl-H2052 cells and NCl-H2452 cells, the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased in NCl-H2052 cells and NCl-H2452 cells ( P<0.001), which was consistent with the results of chip analysis. After exposure of Met-5A cells, it was found that compared with the control group, the expression levels of Vimentin and Twist genes, hsa-miR-155-5p, hsa-miR-34b-5p and hsa-miR-34c-5p in the crocidolite group were increased, while the expression level of E-cadherin gene was decreased ( P<0.05). Compared with the control group, the expression levels of Vimentin, Twist and E-cadherin genes in chrysotile group were increased, while the expression levels of hsa-miR-34b-5p, hsa-miR-34c-5p and hsa-miR-28-5p were decreased ( P<0.05) . Conclusion:Long-term exposure to chrysotile and crocidolite could cause Met-5A cells to produce miRNAs and EMT-related gene expression changes similar to mesothelioma cells.

Objective To analyze the acupoint selection rule for cancer-related fatigue with acupoint stimulation using data mining techniques.Methods Clinical randomized controlled trial research literature about treatment for cancer-related fatigue with acupoint stimulation was retrieved from CNKI,Wanfang Data,VIP,SinoMed,PubMed,Web of Science and Embase from establishment of the databases to October 13,2023.The frequency anlysis,association rule analysis and clustering analysis on acupoints were performed using Excel 2021 and R 4.3.1.Results Totally 187 articles were included,with 214 combinations of acupoints involving 102 acupoints,with a total frequency of 1 044.The most frequently used acupoints were Zusanli(ST36),Qihai(CV6),and Guanyuan(CV4).The top three meridians used were the Conception Vessel,stomach meridian,and spleen meridian.The acupoints were mainly distributed in the lower limbs and thoracoabdominal,and the most commonly used specific acupoint was the crossing point.Moxibustion was the most commonly used intervention measure.Association rule analysis indicated that the core combination of acupoints in this study was Zusanli(ST36),Qihai(CV6),Guanyuan(CV4),and Sanyinjiao(SP6).Analysis of acupoint selection for cancer-related fatigue with different types of cancer demonstrated that acupoints with tonifying properties were most frequently chosen.Clustering analysis identified five effective combinations of acupoints.Conclusion The acupoint selection for treating cancer-related fatigue should focus on tonifying qi and nourishing blood,as well as supporting healthy qi to eliminate pathogenic factors.Special attention is given to the use of specific acupoints,and various acupoint combination methods are employed based on the differentiation of zang-fu organs and meridians to enhance clinical efficacy.

Objective To investigate the clinical value of fiber nasopharyngoscope applied in adenoid hypertrophy(AH)combined with allergic rhinitis(AR)in children.Methods Clinical data of 174 pediatric patients from January 2021 to March 2024 was collected and analyzed.Among them,129 cases were diagnosed with AH via fiber nasopharyngoscope examination(79 cases with AR were assigned to the AH with AR group,the remaining 50 cases of simple AH without AR were assigned to the AH group),and 45 cases of simple AR without AH through fiber nasopharyngoscope examination were assigned to the AR group.And 25 healthy children who came to our pediatric health department for health examinations during the same period were selected as the healthy control(HC)group.On the day of admission,all subjects underwent lateral X-ray examination of the nasopharynx,and the ratio of the maximum thickness of adenoids to the anterior posterior diameter of the nasopharynx cavity(A/N ratio)was calculated.Meanwhile their of peripheral blood eosinophil(EOS)percentage,serum interleukin-17(IL-17),and tumor necrosis factor-α(TNF-α)levels were tested.The A/N ratio,peripheral blood EOS percentage,serum IL-17 and TNF-α levels were compared among the AH with AR group,AH group,AR group,and HC group.The A/N ratio,peripheral blood EOS percentage,serum IL-17 and TNF-α levels of children with different degrees of adenoid obstruction under fiber nasopharyngoscope were compared in AH and AR group.Spearman correlation coefficient was used to analyze the correlation between the degree of adenoid obstruction under fiber nasopharyngoscope and the levels of peripheral blood EOS percentage,serum IL-17 and TNF-α in children from AH and AR group.Result A/N ratio:the value in AH with AR group was higher than that in AH group(P＜0.05),the value in AH group was higher than that in AR group(P＜0.05),and the value in AR group was higher than that in HC group(P＜0.05).Peripheral blood EOS percentage,serum IL-17 and TNF-α levels:AH with AR group had higher levels than those in AR group(P＜0.05),AR group had higher levels than those in AH group(P＜0.05),and AH group had higher levels than those in HC group(P＜0.05).The A/N ratio,peripheral blood EOS percentage,serum IL-17 and TNF-α levels in children with adenoid obstruction degree Ⅲ～Ⅳ under fiber nasopharyngoscope in the AH group were significantly higher than those in children with degree Ⅰ～Ⅱ(P＜0.05).Spearman correlation analysis showed that the degree of adenoid obstruction under fiber nasopharyngoscope in children with AH accompanied by AR significantly positively correlated with peripheral blood EOS percentage,serum IL-17 and TNF-α levels(r values were 0.527,0.451,and 0.402 respectively,P＜0.05).Conclusion Fiber nasopharyngoscope can be used for the diagnosis of AH with AR in children,and can be positive in determining severity of the patient's condition when combined with peripheral blood EOS percentage,serum IL-17 and TNF-α levels.

Objective To explore the acupoint selection law for acupoint stimulation to improve gastrointestinal function after colorectal cancer surgery based on data mining technology.Methods Literature about acupoint stimulation to improve gastrointestinal function was retrieved from CNKI,VIP,Wanfang Data,SinoMed,PubMed,Web of Science and Cochrane Library from the establishment of databases to July 1,2023.Data extraction and analysis were performed using Excel 2021.Association rule analysis and clustering analysis were conducted using R language.Results Totally 197 articles were collected,with 242 combinations of acupoints involving 69 acupoints and the total use frequency of acupoints was 878 times.The top 3 acupoints used in frequency were Zusanli(ST36),Shangjuxu(ST37),and Neiguan(PC6).The top 3 involved meridians were the stomach meridian,Conception Vessel,and spleen meridian.The acupoints were mainly distributed in the lower limbs,and the most commonly used intervention method was needle punching.The association analysis discovered a combination of acupoints centered around Zusanli(ST36),which included Shangjuxu(ST37),Neiguan(PC6),and Sanyinjiao(SP6).Clustering analysis identified 4 effective cluster combinations.Conclusion Acupoint stimulation improves gastrointestinal function after colorectal cancer surgery.Multiple acupoint selection ideas are used,and the summarized acupoint selection and compatibility law in this study can provide reference for clinical research and application.

Objective:To investigate the inhibitory effect of microRNA-106b in the process of migration and invasion of human malignant pleural mesothelioma cell NCI-H2452.Methods:In April 2017, the expression level of miRNA-106b in malignant pleural mesothelioma cells (NCI-H2452, MSTO-211H, NCI-H2052) and normal mesothelial cells MeT-5A was detected and analyzed. Using NCI-H2452 cells as a model, the NCI-H2452 cell model with miRNA-106b overexpression was established by transfecting miRNA-106b mimics. The expression level of miRNA-106b in the cells was detected by real-time fluorescent quantitative PCR. The effect of miRNA-106b on the migration and invasion ability of NCI-H2452 cells was analyzed. The gene expression data of malignant mesothelioma and the downstream target gene data of miRNA-106b in public databases were analyzed to screen the downstream target genes of miRNA-106b in mesothelioma cells that affect cell migration and invasion ability, and to verify the expression of this gene in NCI-H2452 cells with miRNA-106b overexpression.Results:The expression of miRNA-106b in three MPM cells was decreased compared with MeT-5A cells ( P<0.001) . The expression level of miRNA-106b was significantly increased after transfection of miRNA-106b mimics ( P<0.001) . The scratch migration levels of the experimental group were 28.45%±4.37%, 38.12%±4.82% and 50.06%±8.92% at 24h, 31h and 48h, respectively. Compared with the control group, the migration level decreased by 37.48%±2.65%, 49.21%±3.45% and 68.14%±3.81% ( P<0.01) . The number of cell migration and invasion decreased in the experimental group compared with the control group ( P<0.001) . Public databases were used to screen and analyze the possibility that TCF21 gene, as a downstream target gene, could affect the migration and invasion ability of MPM cells. The expression level of TCF21 gene was increased after transfection of miRNA-106b mimics in NCI-H2452 cells ( P=0.009) . Conclusion:MiRNA-106b can inhibit the migration and invasion of NCI-H2452 cells and increase the expression of TCF21 gene.

Objective:To investigate the inhibitory effect of microRNA-106b in the process of migration and invasion of human malignant pleural mesothelioma cell NCI-H2452.Methods:In April 2017, the expression level of miRNA-106b in malignant pleural mesothelioma cells (NCI-H2452, MSTO-211H, NCI-H2052) and normal mesothelial cells MeT-5A was detected and analyzed. Using NCI-H2452 cells as a model, the NCI-H2452 cell model with miRNA-106b overexpression was established by transfecting miRNA-106b mimics. The expression level of miRNA-106b in the cells was detected by real-time fluorescent quantitative PCR. The effect of miRNA-106b on the migration and invasion ability of NCI-H2452 cells was analyzed. The gene expression data of malignant mesothelioma and the downstream target gene data of miRNA-106b in public databases were analyzed to screen the downstream target genes of miRNA-106b in mesothelioma cells that affect cell migration and invasion ability, and to verify the expression of this gene in NCI-H2452 cells with miRNA-106b overexpression.Results:The expression of miRNA-106b in three MPM cells was decreased compared with MeT-5A cells ( P<0.001) . The expression level of miRNA-106b was significantly increased after transfection of miRNA-106b mimics ( P<0.001) . The scratch migration levels of the experimental group were 28.45%±4.37%, 38.12%±4.82% and 50.06%±8.92% at 24h, 31h and 48h, respectively. Compared with the control group, the migration level decreased by 37.48%±2.65%, 49.21%±3.45% and 68.14%±3.81% ( P<0.01) . The number of cell migration and invasion decreased in the experimental group compared with the control group ( P<0.001) . Public databases were used to screen and analyze the possibility that TCF21 gene, as a downstream target gene, could affect the migration and invasion ability of MPM cells. The expression level of TCF21 gene was increased after transfection of miRNA-106b mimics in NCI-H2452 cells ( P=0.009) . Conclusion:MiRNA-106b can inhibit the migration and invasion of NCI-H2452 cells and increase the expression of TCF21 gene.

Background::As a non-invasive and effective diagnostic method for small intestinal bacterial overgrowth (SIBO), wild-use of breath test (BT) has demonstrated a high comorbidity rate in patients with diarrhea-predominant irritable bowel syndrome (IBS-D) and SIBO. Patients overlapping with SIBO respond better to rifaximin therapy than those with IBS-D only. Gut microbiota plays a critical role in both of these two diseases. We aimed to determine the microbial difference between IBS-D overlapping with/without SIBO, and to study the underlying mechanism of its sensitivity to rifaximin.Methods::Patients with IBS-D were categorized as BT-negative (IBSN) and BT-positive (IBSP). Healthy volunteers (BT-negative) were enrolled as healthy control. The patients were clinically evaluated before and after rifaximin treatment (0.4 g bid, 4 weeks). Blood, intestine, and stool samples were collected for cytokine assessment and gut microbial analyses.Results::Clinical complaints and microbial abundance were significantly higher in IBSP than in IBSN. In contrast, severe systemic inflammation and more active bacterial invasion function that were associated with enrichment of opportunistic pathogens were seen in IBSN. The symptoms of IBSP patients were relieved in different degrees after therapy, but the symptoms of IBSN rarely changed. We also found that the presence of IBSN-enriched genera ( Enterobacter and Enterococcus) are unaffected by rifaximin therapy. Conclusions::IBS-D patients overlapping with SIBO showed noticeably different fecal microbial composition and function compared with IBS-D only. The better response to rifaximin in those comorbid patients might associate with their different gut microbiota, which suggests that BT is necessary before IBS-D diagnosis and use of rifaximin.Registration::Chinese Clinical Trial Registry, ChiCTR1800017911.

Objective:To determine a simpler and more practical scoring standard for predicting mucosal histological healing in ulcerative colitis (UC).Methods:From April 11, 2017 to February 8, 2021, 68 UC patients diagnosed with mucosal healing under endoscopy and hospitalized at Department of Gastroenterology, the Tenth People′s Hospital of Tongji University and during the same period 60 healthy individuals who underwent endoscopy for health checkup were retrospectively analyzed. Modified Mayo score and ulcerative colitis endoscopic index of severity (UCEIS), the modified Nancy index and Robarts histopathology index were determined based on the collected clinical data, endoscopic reports and histopathological evaluation. The proportions of neutrophils, eosinophils, and plasma cells in the colonic mucosal lamina propria were calculated. The proportions of activated neutrophils and T cells in the colonic mucosal lamina were calculated according to CD177 and CD40L, respectively. The new clinical and laboratory diagnostic formulas were determined by multivariate logistic regression analysis, the effectiveness of the equations was evaluated by receiver operating characteristic curve (ROC).Results:Among the 68 patients with UC, the modified Mayo score was 0.7 (0.4, 1.1), the UCEIS was 0.5 (0.3, 0.8), the Nancy index was 5.9±3.2, and the Robarts histopathology index was 2.6±1.7. According to multivariate logistic regression analysis, the formula for clinical diagnosis of histological healing was Y1=-21.09+ 355.9 X1+ 305.8 X2+ 44.91 X3 ( X1, X2 and X3 were the proportions of neutrophils, eosinophils, and plasma cells, respectively). The results of ROC analysis indicated that Y1<-0.747 was the cut-off value of diagnosis of histological healing, and the area under the curve (AUC) was 0.986 and 95% confidence interval ( CI) was 0.922 to 1.000 ( P<0.001), the sensitivity was 97.10% and the specificity was 91.20%. The formula of laboratory diagnosis of histological healing was Y2=-10.57+ 469.1 X1 + 132.7 X2 + 101.2 X3 + 18.56 X4 ( X1, X2, X3, and X4 were the proportions of CD177 + neutrophils, eosinophils, CD40L + T cells and plasma cells, respectively). The results of ROC analysis indicated that Y2<1.960 was the cut-off value of diagnosis of histological healing, and the AUC was 0.980, 95% CI was 0.913 to 0.999 ( P<0.001), the sensitivity was 84.78%, and the specificity was 100.00%. The new clinical and laboratory diagnostic criteria were positively correlated with the Nancy histological index ( r=0.411 and 0.308, P=0.001 and 0.011), and Robarts histopathology index ( r=0.311, 0.273, P=0.010 and 0.024). Conclusions:Compared with the Nancy index, the new clinical and laboratory diagnostic criteria are simpler and more practical. The new clinical diagnostic formula Y1<-0.747 and the new laboratory diagnosis formula Y2<1.960 are the independent factors for predicting histological healing in UC patients.

Objective:To investigate the survival and death risk factors of mesothelioma cases stratified by the expression levels of CD8 and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) , providing new clue to evaluate disease progression and clinical outcome.Methods:This was a retrospective case report, which included 47 clinically and pathologically confirmed mesothelioma cases on November 2016. Their clinical and pathological information, asbestos exposure history and survival data were collected. Infiltrated lymphocyte, 5-methylcytosine (5-mC) , CTLA-4, CD8 and Ki-67 antigen were detected using hematoxylin-eosin staining and immunohistochemistry. Survival time and death risk factors of mesothelioma patients with different CD8 and CTLA-4 protein expression characteristics were analyzed. And analyze the influence of Ki-67 expression on the survival of patients with different CD8 and CTLA-4 protein and gene expression characteristics.Results:Among the 47 cases, 63.8% (30/47) had low/medium level of infiltrated lymphocyte. The immunohistochemistry scores of CTLA-4, CD8, 5-mC and Ki-67 were 92.97 (54.95, 120.65) , 72.41 (36.62, 89.82) , 11.09 (3.40, 52.89) and 5.88 (2.41, 11.48) , respectively. Patients with CD8 high CTLA-4 high had higher 5-mC level than those with CD8 high CTLA-4 low ( P<0.01) . The median survival time of 27 cases was 0.83±0.29 year. The median survival times of those with CD8 high CTLA-4 high and CD8 high CTLA-4 low were 0.58±0.51 year and 0.83±0.30 year, respectively ( P=0.521) . The immunohistochemistry score of Ki-67 ≥5.88 was an independent death risk factor for patients with CD8 high CTLA-4 low ( HR=8.40, P=0.01) . Under different CD8 and CTLA-4 protein expression characteristics, in the patients with CD8 high CTLA-4 low, the median survival times of those with high and low Ki-67 expression were 0.57±0.11 years and 2.31±0.46 years, respectively ( P<0.01) . Under different CD8 and CTLA-4 mRNA expression characteristics, in the patients with CD8 high CTLA-4 low, the median survival times of those with high and low Ki-67 mRNA expression were 1.20±0.36 years and 3.38±0.43 years, respectively ( P=0.018) . Conclusion:Mesothelioma case with high CD8 but low CTLA-4 content might coexist DNA hypomethylation. In the presence of high Ki-67 expression, their survival time appears to be shortened with increased death risk.