Objective To investigate the effect of long non-coding ribonucleic acid cancer susceptibility can-didate gene 19(lncRNA CASC19)regulating the microRNA(miR)-490-3p/high mobility group protein A2(HMGA2)axis on the proliferation,migration,and chemotherapy resistance of colorectal cancer(CRC)cells.Methods Human normal colonic epithelial cells(FHC)and CRC cell lines(LOVO,HCT116,SW480)were cultured.LOVO cells were randomly divided into Control group,sh-NC group,sh-CASC19 group,sh-CASC19+anti-NC group and sh-CASC19+anti-miR-490-3p group.The lncRNA CASC19,miR-490-3p and HMGA2 mRNA expression were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The relationship between lncRNA CASC19 and miR-490-3p and between miR-490-3p and HM-GA2 were detected by Dual luciferase assay.The cell proliferation ability was detected by cell counting kit-8(CCK-8)assay and colony formation assay.The cell migration ability was detected by cell scratch assay.The cell invasion ability was detected by Transwell assay.The cell metastasis-related proteins(MMP2,MMP9)and HMGA2 protein expression were detected by Western blot assay.The chemotherapy resistance was detec-ted by cisplatin and fluorouracil.Results The lncRNA CASC19 and HMGA2 mRNA expression increased in CRC cell lines,and miR-490-3p expression decreased.The lncRNA CASC19,miR-490-3p and HMGA2 mRNA expression in LOVO cells were the most significant,LOVO cells were selected for subsequent experiments.The dual luciferase assay showed that,after the transfection of lncRNA CASC19 and HMGA2,compared with mimic-NC group,the luciferase activity in miR-490-3p mimic group decreased(P＜0.05).Compared with sh-NC and Control groups,the survival rate,clone number,migration rate,invasion rate,MMP2,MMP9,HMGA2 mRNA and protein expression of LOVO cells in sh-CASC19 group were decreased,and the miR-490-3p ex-pression was increased(P＜0.05).Compared with sh-CASC19 group and sh-CASC19+anti-NC group,the survival rate,clone number,migration rate,invasion rate,MMP2,MMP9,HMGA2 mRNA and protein expres-sion of LOVO cells in sh-CASC19+anti-miR-490-3p group were increased,and the miR-490-3p expression was decreased(P＜0.05).The chemotherapy resistance experiment showed that,compared with Control group,the chemotherapy resistance sensitivity of LOVO cells in sh-CASC19 group increased(P＜0.05).Compared with sh-CASC19 group,the chemoresistance sensitivity of LOVO cells in sh-CASC19+anti-miR-490-3p group was decreased(P＜0.05).In the same group,with the increase of the concentration of fluorou-racil and cisplatin,the cell survival rate gradually decreased(P＜0.05).Conclusion Knockdown of lncRNA CASC19 can regulate miR-490-3 p/HMGA2 signaling pathway,inhibit the proliferation and migration of CRC cells,and increase the sensitivity of chemotherapy resistance.

Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Objective To obtain the recombinant Argonaute protein(TtAgo)from Thermus thermophilus using gene cloning and pro-tein purification techniques and detect its endonuclease activity against the KRAS 12D mutation in vitro,thereby to provide experimental evidence for its subsequent application in the detection of gene mutations.Methods The TtAgo gene sequence was amplified from the genome of Thermus thermophilus by the PCR technology.Then,the amplified sequence was inserted into the prokaryotic expression vec-tor pET-28a to construct the recombinant vector pET-28a-TtAgo.After the recombinant TtAgo protein was induced to be expressed,it was purified sequentially by the nickel ion affinity chromatography(Ni-NTA),heparin affinity chromatography,and size-exclusion chromatography.The high-specificity nuclease activity of the recombinant TtAgo protein mediated by a short DNA guide strand(gDNA)was verified in vitro.Results The prokaryotic expression vector pET-28a-TtAgo was successfully constructed by the gene cloning tech-nique,and the soluble expression of the recombinant TtAgo protein was achieved.With the help of protein purification technology,the high-purity recombinant TtAgo protein was obtained.Under the mediation of gDNA,the recombinant TtAgo protein could specifically cleave wild-type single-stranded DNA(ssDNA),double-stranded DNA(dsDNA),and RNA molecules,while had no cleavage activi-ty for tumor KRAS 12D mutant molecules.Conclusion The high-purity recombinant TtAgo protein with nuclease activity is successful-ly obtained,and its targeted cleavage activity towards the KRAS 12D mutation site is detected in vitro.

OBJECTIVE To explore the effect and potential mechanisms of berberine on formation of biofilms of clinical methicillin-resistant Staphylococcus aureus(MRSA)isolates.METHODS Totally 95 clinical MRSA iso-lates were collected from Shanghai Eighth People's Hospital from Jan.2023 to Dec.2023.The 14 biofilm forma-tion-related genes in the strains were detected by polymerase chain reaction(PCR)and multiplex PCR,the mini-mum inhibitory concentration(MIC)of berberine was determined by microbroth dilution method,the effect of berberine on resistance of biofilm formation was evaluated by crustal violet staining,fluorescence microscope,Congo red agar plate and extracelluar DNA(eDNA).The transcription levels of 9 biofilm formation-related genes were detected by real-time fluorescent quantitative reverse transcription PCR.RESULTS All of the 95 strains of MRSA carried eno,clfA,clfB and icaA genes,the most widespread gene profile was bbp-eno-ebpS-fnbA-fib-clfA-clfB-icaA-sasG,and 29 strains had the phenotypes with strong capability of biofilm formation.The MIC score of berberine ranged between 64 and 1 024 μg/ml.Berberine with the concentration of 1/2 MIC could inhibit the biofilm formation of MRSA(P＜0.001),and the inhibiting rate of biofilm formation of the MIC ≥512 μg/ml group was higher than that of the MIC≤128 μg/ml group and the MIC 256 μg/ml group(all P＜0.05).The re-sult under the fluorescence microscope showed that the fluorescence intensity of biofilms decreased with the rise of berberine concentration.Berberine could reduce the formation of amyloid fibrils and the release of eDNA,down-regulating the transcription levels of ica A,sasG,ebpS,fib,eno,clfA,clfB,bbp and fnbA genes(P＜0.05).CONCLUSION Berberine may inhibit the biofilm formation of MRSA by downregulating expression levels of related genes,interfering the formation of amyloid fibrils and blocking the release of eDNA,which may provide experimental bases for development of drugs resisting to MRSA biofilms.

Objective:To analyze the biological characteristics and genomic features of a Streptococcus pyogenes strain isolated from the pus of a child with an atypical brain abscess. Methods:The Streptococcus pyogenes strain, named 21SPY7071 ( emm22 type), was isolated from the brain abscess specimen of a child undergoing surgery for brain abscess. The biological characteristics of this strain were analyzed using blood agar culture, Gram staining, growth curve measurement, and hemolytic analysis. After whole-genome sequencing, core genome multilocus sequence typing (cgMLST) was performed to analyze the cgST type of the strain, and the distribution of virulence genes and drug resistance genes were analyzed. qRT-PCR was used to detect the differences in the expression of virulence genes at mRNA level between the strain 21SPY7071 and two Streptococcus pyogene strains of ATCC 19615 ( emm80 type) and GAS029 ( emm22 type, clinical isolate). Results:The strain 21SPY7071 was a Gram-positive Streptococcus with atypical weak β-hemolysis. Whole-genome sequencing and cgMLST revealed that this isolate belonged to the cgST type of 65937, containing 187 virulence genes and 97 drug resistance genes, with high sequence similarity (95.16%~100.00%) to the main virulence factor-encoding genes in the Virulence Factor Database. Compared with the strains of ATCC 19615 and GAS029, the strain 21SPY7071 showed reduced expression of genes encoding streptolysin S ( sagABCDEFGHI), streptolysin O ( slo), and hyaluronate lyase ( endA/ sdaB). Besides, compared with the strain ATCC 19615, the strain 21SPY7071 also showed decreased expression of virulence genes including speG, ideS/ mac, hylA, smeZ, lmb, and scpA/ scpB. Conclusion:This Streptococcus pyogenes strain isolated from the pus of a child with atypical brain abscess exhibits weak hemolysis and lower expression of virulence genes, which may be related to the mild clinical symptoms observed in the child.

Objective:To investigate the effects of lactulose and oral rehydration salt powder (Ⅲ) on bowel preparation for colonoscopy and their impact on the detection rate of lesions.Methods:A retrospective study was conducted with 120 patients who underwent colonoscopy at The Second Affiliated Hospital of Wenzhou Medical University from January 2023 to April 2024. The patients were randomly divided into a control group and an experimental group, with 60 patients in each group. The control group received Hengkang Zhengqing and simethicone for bowel preparation, while the experimental group received lactulose, oral rehydration salt powder (Ⅲ), and simethicone for bowel preparation. The Boston Bowel Preparation Scale (referred to as the Boston scale) was used to assess bowel cleanliness, the Bubble scale was used to assess the presence of gas in the bowel, and a self-designed questionnaire was used to evaluate patient satisfaction after colonoscopy. The Boston scale scores, Bubble scale scores, patient satisfaction, lesion detection rates, and the incidence of adverse reactions were compared between the two groups. Results:The total score on the Boston scale in the experimental group was significantly higher than that of the control group [(7.23 ± 1.30) vs. (5.44 ± 1.17), t = 7.92, P < 0.001]. The Bubble scale score in the experimental group was significantly higher than that of the control group [(2.60 ± 0.15) vs. (2.21 ± 0.10), t = 16.75, P < 0.001]. The rates of acceptable taste and tolerance in the experimental group were 96.6% (58/60) and 95.0% (57/60), respectively, both of which were higher than those in the control group at 86.6% (52/60) and 81.6% (49/60) ( χ2 = 3.92, 5.17, P = 0.048, 0.023). The detection rate of colonic polyps in the experimental group was significantly higher than that in the control group [31.67% (19/60) vs. 15.00% (9/60), χ2 = 4.65, P = 0.031]. The incidence of adverse reactions in the experimental group was significantly lower than that in the control group [3.33% (2/60) vs. 13.33% (8/60), χ2 = 3.92, P = 0.048]. The patient satisfaction rate after examination in the experimental group was significantly higher than that in the control group [96.67% (58/60) vs. 85.00% (52/60), χ2 = 4.90, P = 0.027]. Conclusions:The intestinal preparation for colonoscopy using the lactulose and oral rehydration salt powder (Ⅲ) regimen can significantly improve the quality of bowel preparation and the detection rate of lesions. Additionally, it offers better taste, higher patient tolerance, and greater satisfaction, along with fewer adverse reactions compared with the Hengkang Zhengqing and simethicone regimen.