Objective:To investigate the molecular mechanisms of chronic rhinosinusitis (CRS), to identify key cell subgroups and genes, to construct effective diagnostic models, and to screen for potential therapeutic drugs.Methods:Key cell subgroups in CRS were identified through single-cell transcriptomic sequencing data. Essential genes associated with CRS were selected and diagnostic models were constructed by hdWGCNA (high dimensional weighted gene co-expression network analysis) and various machine learning algorithms. Causal inference analysis was performed using Mendelian randomization and colocalization analysis. Potential therapeutic drugs were identified using molecular docking technology, and the results of bioinformatics analysis were validated by immunofluorescence staining. Graphpad Prism, R, Python, and Adobe Illustrator software were used for data and image processing.Results:An increased proportion of basal and suprabasal cells was observed in CRS, especially in eosinophilic CRS with nasal polyps (ECRSwNP), with P=0.001. hdWGCNA revealed that the "yellow module" was closely related to basal and suprabasal cells in CRS. Univariate logistic regression and LASSO algorithm selected 13 key genes ( CTSC, LAMB3, CYP2S1, TRPV4, ARHGAP21, PTHLH, CDH26, MRPS6, TENM4, FAM110C, NCKAP5, SAMD3, and PTCHD4). Based on these 13 genes, an effective CRS diagnostic model was developed using various machine learning algorithms (AUC=0.958). Mendelian randomization analysis indicated a causal relationship between CTSC and CRS (inverse variance weighted: OR=1.06, P=0.006), and colocalization analysis confirmed shared genetic variants between CTSC and CRS (PPH4/PPH3>2). Molecular docking results showed that acetaminophen binded well with CTSC (binding energy:-5.638 kcal/mol). Immunofluorescence staining experiments indicated an increase in CTSC +cells in CRS. Conclusion:This study integrates various bioinformatics methods to identify key cell types and genes in CRS, constructs an effective diagnostic model, underscores the critical role of the CTSC gene in CRS pathogenesis, and provides new targets for the treatment of CRS.

Objective To detect the relative expression level of long non-coding RNA nuclear enriched a-bundant transcript(NEAT)1 in peripheral serum of patients with gastric cancer,and to explore the possibili-ty of its application in the auxiliary diagnosis of gastric cancer.Methods A total of 101 patients with newly diagnosed gastric cancer in Nantong Tumor Hospital from November 2017 to December 2019 were selected as the gastric cancer group,60 patients with benign gastric lesions were selected as the gastric benign lesions group,and 76 healthy people were selected as the control group.The relative expression levels of lncRNA NEAT1,carcinoembryonic antigen(CEA)and carbohydrate antigen 19-9(CA19-9)in serum samples of each group were detected,and the correlation between lncRNA NEAT1 and clinicopathological parameters was ana-lyzed.Receiver operating characteristic(ROC)curve was used to evaluate the diagnostic efficacy of lncRNA NEAT1,CEA and CA19-9 alone or in combination.Results The relative expression levels of lncRNA NEAT1 in serum of gastric cancer group,gastric benign disease group and control group were 2.163(1.357,2.843),1.176(0.677,1.381)and 1.063(0.570,1.308),respectively,and the difference was statistically significant(H=52.467,P＜0.001).The relative expression level of serum lncRNA NEAT1 in gastric cancer patients was not related to gender(P=0.353)and age(P=0.382),but was related to tumor size(P=0.031),TNM stage(P=0.037)and lymph node metastasis(P=0.046).The relative expression level of lncRNA NEAT1 in serum of gastric cancer patients was not correlated with CEA and CA19-9(P＞0.05).The ROC curve showed that the area under the curve and sensitivity of lncRNA NEAT1 in the diagnosis of gastric cancer were higher than those of CEA and CA19-9.The combined diagnosis of the three had the highest diagnostic efficiency.Conclusion The relative expression of lncRNA NEAT1 in peripheral blood of patients with gastric cancer is increased,and it is correlated with some clinicopathological parameters.lncRNA NEAT1 may be a new marker for auxiliary diagnosis of gastric cancer.

Gastric cancer(GC)is one of the most common gastrointestinal tumors.As a newly discovered type of non-coding RNAs,transfer RNA(tRNA)-derived small RNAs(tsRNAs)play a dual biological role in cancer.Our previous studies have demonstrated the potential of tRF-23-Q99P9P9NDD as a diagnostic and prognostic biomarker for GC.In this work,we confirmed for the first time that tRF-23-Q99P9P9NDD can promote the proliferation,migration,and invasion of GC cells in vitro.The dual luciferase reporter gene assay confirmed that tRF-23-Q99P9P9NDD could bind to the 3'untranslated region(UTR)site of acyl-coenzyme A dehydrogenase short/branched chain(ACADSB).In addition,ACADSB could rescue the effect of tRF-23-Q99P9P9NDD on GC cells.Next,we used Gene Ontology(GO),the Kyoto Encyclopedia of Genes and Genomes(KEGG),and Gene Set Enrichment Analysis(GSEA)to find that downregulated ACADSB in GC may promote lipid accumulation by inhibiting fatty acid catabolism and ferroptosis.Finally,we verified the correlation between ACADSB and 12 ferroptosis genes at the transcriptional level,as well as the changes in reactive oxygen species(ROS)levels by flow cytometry.In summary,this study proposes that tRF-23-Q99P9P9NDD may affect GC lipid metabolism and ferroptosis by targeting ACADSB,thereby promoting GC progression.It provides a theoretical basis for the diagnostic and prognostic monitoring value of GC and opens up new possibilities for treatment.

Objective:To investigate the correlation between FCER2（2206A>G） gene polymorphism and the efficacy of inhaled corticosteroids（ICS） in patients with chronic rhinosinusitis（CRS）. Methods:A total of 208 CRS patients were routinely treated with functional endonasal sinus surgery and postoperative ICS. DNA extraction, PCR amplification and gene sequencing were performed to observe the FCER2（2206A>G） gene polymorphism and calculate the allele frequency. The visual analog scale（VAS） score, Lund-Kennedy score, and computed tomography（CT） Lund-Mackay score were determined 6 months after surgery among patients with different genotypes. Moreover, the polymorphism frequency was compared among different subgroups（chronic rhinosinusitis with nasal polyps versus chronic rhinosinusitis without nasal polyps, eosinophilic chronic rhinosinusitis versus non-eosinophilic chronic rhinosinusitis）. Results:There were FCER2（2206A>G） gene polymorphism in patients with CRS, and the phenotypes included 3 genotypes, AA, AG and GG, with distribution frequencies of 68（32.7%）, 116（55.8%） and 24（11.5%） cases, respectively. No significant differences were found in age, VAS score, nasal endoscopic Lund-Kennedy score and CT imaging Lund-Mackay score among patients with CRS of each genotype before surgery. In patients with the AA genotype, the changes in VAS score（5.74±1.10）, Lund Kennedy score（5.92 ± 1.14）, and CT imaging Lund-Mackay score（13.26±4.26） were significantly higher than in patients with the AG（4.37±0.86, 5.37±1.24, 10.82±3.77） and GG（4.26±0.80, 5.18±1.56, 10.10±3.53） genotype（P<0.05）. However, there were no marked difference between patients with the AG genotype and those with the GG genotype（P>0.05）. Compared with patients with non-eosinophilic sinusitis, Among them, the differences between the GG genotype and AG /AA genes were more significant in eosinophilic sinusitis compared to non-eosinophilic sinusitis（P<0.01）. Conclusion:The FCER2（2206A>G） gene in patients with CRS has genetic polymorphism and is associated with the recovery of CRS patients after surgery, individual corticosteroid sensitivity, and subgroup variability.
Humans ; Nasal Polyps/complications* ; Rhinitis/complications* ; Sinusitis/complications* ; Adrenal Cortex Hormones/therapeutic use* ; Polymorphism, Genetic ; Endoscopy/methods* ; Chronic Disease ; Receptors, IgE ; Lectins, C-Type

Objective:To investigate the expression of hsa_circ_0000437 in the serum of patients with gastric cancer and its clinical value.Methods:The serum samples from 80 patients (57 males and 23 females) with pathologically confirmed gastric cancer (GC), 50 gastric benign disease (28 males and 22 females) and 80 healthy controls (46 males and 34 females) were collected from October 2018 to December 2020 in Affiliated Hospital of Nantong University.Serum samples from 35 of 80 gastric cancer patients after operation were collected. The expression of serum hsa_circ_0000437 was determined by real-time fluorescent quantitative PCR (RT-qPCR). Serum carcinoembryonic antigen (CEA), carbohydrate antigen 199 (CA 199) and carbohydrate antigen 724 (CA724) were determined by chemiluminescence method.Comparisons of serum hsa_circ_0000437 between groups were performed by Mann-Whitney U test.The correlation between serum expression of hsa_circ_0000437 in gastric cancer patients and its clinical pathological characteristics was performed by χ 2 test.Receiver operating characteristic (ROC) curve and the area under the curve of ROC (AUC) were used to evaluate their diagnosis efficiency. Kaplan-Meier survival curve analysis was used to analyze the relationship between the expression level of serum hsa_circ_0000437 and the prognosis of patients. Results:The relative expression of hsa_circ_0000437 in GC, gastric benign disease, healthy controls were 2.252 (1.235, 4.765), 1.598(1.139, 1.982) and 1.000 (0.818, 1.385) respectively.The relative expression of hsa_circ_0000437 in GC was significantly higher than that in gastric benign disease ( P<0.001) and healthy controls ( P<0.001). The difference between gastric benign disease and healthy controls was also statistically significant ( P<0.001).The differences of serum hsa_circ_0000437 expression in GC patients between T stage, N stage, and tumor differentiation were statistically significant. The AUC of hsa_circ_0000437, CEA, CA199 and CA724 in GC patients were 0.863, 0.619, 657 and 0.608 respectively compared with healthy controls. The AUC of above four-parameter panel was 0.892 and the sensitivity was up to 97.5% (78/80). Kaplan-Meier survival curve showed that the overall survival rate of patients with high serum hsa_circ_0000437 expression was significantly lower than that of patients with low expression ( P=0.008). Conclusion:Serum hsa_circ_0000437 could be a biomarker for the auxiliary diagnosis and prognosis of GC.

At present, nucleic acid testing technology has been widely used in clinical laboratory diagnosis. Conventional detection technique such as real-time PCR is complicated, time consuming, and dependent on specific instruments. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas) system is an adaptive immune defense system against viruses in bacteria and archaea, which has been developed into a powerful technology for genome editing. Recently, the leading groups engaged in CRISPR have set up new tools for nucleic acid detection based on Cas13a, Cas12a and newly discovered protein-Cas14, which plays an important role in rapid diagnosis of infectious diseases, detection of gene mutations in cancer and genotyping. Since they are ultrasensitive, specific, rapid and cost-effective, it is expected to bring great potential for molecular diagnosis. In this review, the mechanism of CRISPR/Cas system and the principle, the applications of the newly-developed diagnostic platforms are introduced. What′s more, the advantages, limitations and prospects of the technologies are summarized.

Objective:To investigate the expression and its diagnostic value of long-chain non coding RNA (lncRNA) and colon cancer-related transcript-2 (CCAT2) in serum of patients with cervical cancer (CC).Methods:Serum samples of 100 CC patients, 60 CIN patients and 80 healthy people enrolled by Nantong Tumor Hospital from January 2016 to June 2017 were collected.The expression levels of CCAT2 in sera of CC patients and their corresponding postoperative patients, CIN patients and healthy controls were detected by real-time fluorescent quantitative PCR (RT-qPCR). The correlation between CCAT2 and clinicopathological features, as well as the traditional auxiliary diagnostic makers of CC, such as carbohydrate antigen 125 (CA125) and squamous cell carcinoma antigen (SCC) was analyzed. The working characteristic curve (ROC) of the subjects was used to evaluate the CCAT2 pair in the auxiliary diagnostic value of cervical cancer.Results:The relative expression levels of serum CCAT2 in patients with cervical cancer, patients after operation, patients with CIN and healthy controls were1.689 (0.616, 4.776), 1.018 (0.227, 3.328), 0.815 (0.453, 1.266) and 0.740 (0.271, 1.670), respectively.The relative expression of CCAT2 in cervical cancer patients was significantly higher than that in post-operative patients, CIN patients and healthy controls. The difference was statistically significant( t=6.999,8.193,9.345, P<0.001).There was no significant difference in the relative expression of CCAT2 between CIN patients and healthy controls ( t=0.327, P>0.05).The relative expression of CCAT2 in serum of cervical cancer patients had no significant difference in age 1.636(1.000,2.370),1.705(1.095,2.243) (χ 2=0.137, P=0.712) and menopause 1.672(1.059,2.342),1.659(1.068,2.298) (χ 2=0.000, P=1.000), but had significant difference with tumor size expression1.189(0.916,1.725),2.019(1.537,2.497)(χ 2=17.508, P=0.000),International Federation of Obstetrics and Gynecology (FIGO) staged expression stage 0.993(0.779,1.266),2.056(1.547,2.549),3.987(3.699,4.275)(χ 2=36.075, P=0.000) and lymph node metastasis 1.434(1.007,2.251),2.019(1.731,3.098) (χ 2=8.634, P=0.003). There was no correlation between the relative expression of serum CCAT2 and CA125 ( r2=0.003, P=0.563) and SCC (r 2=0.128, P=0.000).The diagnostic efficacy of serum CCAT2, CA125 and SCC in cervical cancer patients was analyzed by ROC curve. When compared with CIN patients, the areas under the curve were 0.890, 0.549 and 0.744, respectively. When compared with healthy patients, the areas under the curve were 0.857, 0.650 and 0.758, respectively. Conclusions:The level of serum CCAT2 in patients with cervical cancer is significantly higher than that in patients with cervical cancer, CIN patients and healthy controls. Serum CCAT2 may be a relevant marker for the diagnosis and prognosis of cervical cancer.

Objective:To explore the expression level of serum exosomal miR-223-3p in patients with multiple myeloma (MM) and evaluate its clinical application.Methods:A case-control study was performed. Serum samples were collected from the Department of Hematology, Affiliated Hospital of Nantong University from March 2018 to August 2019, including 68 newly diagnosed untreated MM patients (16 in stage Ⅰ, 22 in stage Ⅱ and 30 in stage Ⅲ), 56 healthy controls and 30 patients with recurrent MM in the same period, and follow-up patients who underwent chemotherapy for 3 months after initial diagnosis. Serum exosomes were purified and identified, RNA was extracted, and RT-qPCR was performed to compare the levels of miR-223-3p in serum exosomes between the groups. The correlation between exosomal miR-223-3p levels and clinical pathological data was analyzed, and the diagnostic efficacy was evaluated by ROC curve.Results:Compared with healthy controls (0.92±0.29), the expression level of serum exosomal miR-223-3p in newly diagnosed MM patients (0.52±0.23) decreased significantly ( U=565, P<0.000 1). According to the international staging system of MM, the expression level of serum exosomal miR-223-3p in patients with stage Ⅰ(0.67±0.26) was significantly higher than that in patients with stage Ⅱ (0.47±0.21) and Ⅲ (0.48±0.19) ( U values were 96.5 and 138 respectively, P were all less than 0.05). In addition, the expression level of serum exosomal miR-223-3p increased after 3 months of chemotherapy (0.69±0.19) compared with that before chemotherapy (0.50±0.22) ( U=228.5, P=0.000 8). Compared with the healthy control group (0.84±0.21), the expression level of serum exosomal miR-223-3p in recurrent MM patients (0.65±0.18) was also decreased ( U=244, P=0.002). At the same time, the expression level of serum exosomal miR-223-3p may be related to the low expression of albumin ( U=411.5, P=0.043 1) and bone damage ( U=309, P=0.001). The ROC curves showed that the AUC of serum exosomal miR-223-3p in the diagnosis of MM was 0.853, which was higher than that of β 2-microglobulin (β 2-MG) (AUC=0.805) and albumin (AUC=0.743). The AUC of the three combined detection was 0.918. Conclusions:Serum exosomal miR-223-3p can be used as a potential marker for auxiliary diagnosis of MM, and its combination with β 2-MG and albumin can improve the diagnostic efficacy of MM.

Objective: To investigate the expression of circular RNA hsa_circ_0128298 in hepatocellular harcinoma (HCC) and evaluate its function in the growth process of HCC. Methods: qPCR was used to detect the expressions of hsa_circ_0128298 in 50 HCC tissues, corresponding paracancerous tissues and HCC cells. The effects of hsa_circ_0128298 on HCC growth was analyzed by CCK-8 kit, flow cytometry and western blot assays. The values of hsa_circ_0128298 for the diagnosis and prognosis of HCC were assessed by ROC curve and survival curve analyses. Results: Compared with the paracancerous tissues, the expression of hsa_circ_0128298 was significantly increased in HCC tissues ( U =846.0， P =0.005). The area under ROC curve (AUCROC) was 0.661, 95% confidence interval ( CI ) was 0.560 to 0.753, sensitivity was 80.0% and specificity was 50.0%. HCC patients with high expression of hsa_circ_0128298 displayed less overall survival time than those with low expression of hsa_circ_0128298 ( χ 2=6.294, P =0.012). RNA interference of hsa_circ_0128298 (si-hsa_circ_0128298) significantly inhibited HCC cell proliferation and induced cell cycle arrest at G0/G1, and also induced mRNA upregulation and protein expression of cyclin p21. Meanwhile，si-p21 partially reversed the proliferation inhibition and cell cycle arrest of HCC cells induced by si-hsa_circ_0128298, and restore its growth and proliferation potential. Conclusion The highly expressed hsa_circ_0128298 in HCC could promote the growth of HCC by regulating the expression of p21 and promise to be a new target for diagnosis and treatment of HCC.

Objective To analyze the identification, drug resistance and clinical significance of a rare bacterium of Bordetella holmesii ( B. holmesii) to improve its detection and clinical diagnosis and treat-ment. Methods A strain isolated from a bacteremia case was identified by bacterial culture, biochemical tests and 16S rRNA gene sequencing. Mega 7. 0 software was used to conduct a similarity analysis of 16S rRNA gene sequences between the type strains of Bordetella spp. and the isolate, and then a phylogenetic tree was constructed. Antibiotic resistance of the isolate was determined by E-test. Changes in bacterial growth were measured after adding different concentrations of riboflavin or its inhibitor lumiflavin to the cul-ture medium. Results B. holmesii ABD2 was the pathogen causing bacteremia in the immunocompetent pa-tient. It was deposited under the number of CGMCC 1. 13721 in China General Microbiological Culture Col-lection Center (CGMCC), and the 16S rRNA gene sequences were deposited in National Center for Biotech-nology Information ( NCBI) with the accession number of KT828544. 1. Unrooted tree showed that the B. holmesii strain was highly homologous with B. pertussis. Antibiotic susceptibility test showed that the mini-mum inhibitory concentrations ( MIC) of piperacillin, ceftazidime, cefepime, imipenem, meropenem, cipro-floxacin, levofloxacin, gentamicin, amikacin, erythromycin, tetracycline and polymyxin B against the isolate were low, while the MIC values of cefazolin, cefuroxime, cefoxitin, cefotaxime, aztreonam and trime-thoprim-sulfamethoxazole were high. Riboflavin accelerated the growth of B. holmesii ABD2, while its inhibi-tor lumiflavin had an inhibitory effect. Conclusions As B. holmesii is hard to isolate and identify, limited clinical, microbiological and epidemiological data are available. It is an under-recognized pathogen with a considerable amount of information that remains to be studied.