ObjectiveTo investigate the ability of clinically isolated， Helicobacter pylori （H. pylori）-specific gene polymerase chain reaction （PCR）-positive gastric， vaginal， and fecal Candida to release H. pylori. MethodsResuscitate 4 strains of H. pylori -specific 16S rDNA and ureA gene PCR-positive Candida strains isolated in laboratory from clinical sources， including 1 strain of gastric Candida， 1 strain of fecal Candida， 2 strains of vaginal Candida and the standard Candida albicans strain ATCC10231 （Ca10231）. The presence of H. pylori-specific ureA in the 5 strains of Candida isolates was confirmed by PCR. The aforementioned strains of Candida and H.pylori were inoculated into urea medium and cultured in a constant temperature incubator at 37 ℃. The color change of the medium was observed daily. A change in the medium's color from yellow to red indicated the presence of urease activity. Then， the five strains of Candida and H. pylori were co-incubated with the magnetic beads coated with H. pylori antibodies respectively. Scanning electron microscopy （SEM） was employed to observe the presence of bacilli adsorbed on the surface of the magnetic beads. PCR was used to detect the presence of H.pylori-specific 16S rDNA and ureA genes on magnetic beads. ResultsThe PCR analysis of the ureA gene in the four Candida isolates was positive， whereas the Ca10231 strain tested negative. Upon culturing the four Candida isolates on urea medium， the medium color changed from yellow to red which was determined to be urease positive， while the medium containing Ca10231 remained unchanged， which was urease negative. SEM revealed that bacilli could be observed on the surface of magnetic beads co-incubated with the 4 strains of Candida of clinical origin and H.pylori isolate. Specifically， PCR testing of the magnetic beads co-incubated with one vaginal Candida， one gastric Candida and H.pylori isolate showed positive results for the 16S rDNA and ureA genes of H. pylori； however， the PCR tests for the two genes were negative for the magnetic beads co-incubated with the other two Candida isolate. ConclusionThis study demonstrates that H. pylori-specific genes Candida can release H. pylori.

ObjectiveTo investigate the effects of berberine hydrochloride on the expression of urease genes ureA and ureB in Helicobacter pylori （H. pylori） and its biofilm. MethodsThe standard strain 26695 of H. pylori， which was preserved in our laboratory， along with the drug-resistant strains H390， e660， H57， and H389 of H. pylori that were isolated from clinical sources were revived. The minimum inhibitory concentration （MIC） of berberine hydrochloride against the planktonic cells of the aforementioned five strains of H. pylori was determined using the broth dilution method. The combined inhibitory effect of berberine hydrochloride and amoxicillin against the aforementioned four strains of clinically resistant H. pylori was assessed using the checkerboard method. TheMIC of berberine hydrochloride against the biofilms of H. pylori strains 26695 and H390 was determined using the broth dilution method. Planktonic cells and biofilms of the aforementioned H. pylori strains 26695 and H390 were co-incubated with berberine hydrochloride at concentrations of 0.5MIC， MIC， and 2MIC for 2， 8， and 24 hours， respectively. Subsequently， urease activity was measured， and the expression levels of the ureA and ureB genes were detected using RT-qPCR. ResultsBerberine hydrochloride exhibited inhibitory effects against both planktonic cells of the antibiotic-sensitive strain 26695 and the antibiotic-resistant strains H390， e660， H57， and H389 of H. pylori. It also demonstrated inhibitory effects against the biofilms formed by strains 26695 and H390. Furthermore， the combined inhibitory effect of berberine hydrochloride and amoxicillin against antibiotic-resistant strains of H. pylori was either synergistic or additive. Berberine hydrochloride inhibited urease activity in both planktonic cells and biofilms of the H. pylori strains 26695 and H390. Furthermore， berberine hydrochloride downregulated the relative expression levels of the ureA and ureB genes in H. pylori and its biofilms. The relative expression levels of the ureA and ureB genes exhibited a dose- and time-dependent decrease with increasing concentrations of berberine hydrochloride and longer co-incubation times （P<0.05）. ConclusionBerberine hydrochloride inhibits both planktonic cells and biofilms of H. pylori. When combined with amoxicillin， it exerts additive or synergistic effects against H. pylori. Furthermore， by inhibiting urease activity and downregulating the expression of the ureA and ureB genes in H. pylori， berberine hydrochloride contributes to the reduction in growth of both planktonic cells and biofilms.

OBJECTIVES: To explore the mechanism of cinnamic acid (CA) for improving doxorubicin-induced myocardial injury (DIC) in mice. METHODS: Network pharmacology analysis was used to obtain the key targets of CA and DIC. Male C57BL/6J mice were randomized into Sham, DOX, CA (25, 50 and 100 mg/kg)+DOX, and CA+Ferrostatin-1+DOX groups, and their myocardial function and pathology were examined by echocardiography and HE staining. Serum levels of CK-MB, LDH, MDA, IL-6, TNF‑α and myocardial ROS level were detected, and the expression levels of TLR4 and ferroptosis pathway proteins in myocardial tissue were detected by Western blotting. Cultured murine cardiomyocytes (HL-1 cells) with or without transfection with a small interfering RNA targeting TLR4 (si-TLR4) were treated with DOX or Erastin, and the cellular ROS content was measured by DCFH-DA staining; the expression level of GPX4 was detected using immunofluorescence staining. RESULTS: Network pharmacology analysis suggested that CA may improve DIC through TLR4 signaling. DOX treatment caused obvious myocardial injury in mice, which showed significantly increased serum levels of CK-MB, LDH, MDA, IL-6, TNF-α and myocardial ROS level with decreased myocardial levels of SLC7A11 and GPX4 proteins and increased levels of TLR4 and PTGS2 proteins. All these changes in the mouse models were significantly alleviated by treatment with CA, and the mice receiving CA or ferrostatin-1 treatment exhibited increased myocardial expressions of SLC7A11 and GPX4 proteins and lowered expressions of TLR4 and PTGS2 proteins. In cultured HL-1 cells, treatment with DOX and Erastin both obviously increased intracellular ROS level and decreased cellular GPX4 expression level, and these changes were strongly attenuated by TLR4 interference. CONCLUSIONS CA, as a potent herbal monomer, can effectively alleviate DIC in mice by inhibiting TLR4-mediated ferroptosis.
Animals ; Ferroptosis/drug effects* ; Toll-Like Receptor 4/metabolism* ; Myocytes, Cardiac/metabolism* ; Mice, Inbred C57BL ; Mice ; Male ; Doxorubicin/adverse effects* ; Cinnamates/pharmacology* ; Signal Transduction ; Reactive Oxygen Species/metabolism*

This study was aimed at investigating the presence of Helicobacter hepaticus(Hh)infection in patients with digestive tract diseases and evaluating Helicobacter pylori(Hp)infection status in patients with digestive tract cancers other than gastric cancer.Fecal samples were collected from 197 patients with digestive tract diseases at the Affiliated Cancer Hospital of Guizhou Medical Uni-versity and from 149 healthy volunteers residing in Guiyang.Hp stool antigen(HpSA)was detected with the colloidal gold method.Af-ter the extraction of fecal DNA,the Hp specific ureA gene and the Hh specific 16S rRNA gene were amplified via nested PCR,and the amplified products were subsequently confirmed through sequencing analysis.The study included 197 patients with digestive system diseases,comprising 135 cases of colorectal cancer,32 cases of chronic gastritis,22 cases of gastric cancer,5 cases of liver cancer,and 3 cases of cholangiocarcinoma.The detection rate of HpSA was 31.5%(62/197).HpSA was detected across all five disease catego-ries,and the highest detection rate was observed in patients with gastric cancer,at 50.0%(11/22),or colorectal cancer,at 24.4%(33/135).The positivity rate of Hp ureA gene PCR was 7.6%(15/197),and sequencing confirmed that the amplified products were in-deed Hp ureA gene fragments.Notably,the highest detection rate was observed in patients with colorectal cancer,at 8.9%(12/135).The positivity rate of Hh 16S rRNA gene PCR was 11.2%(22/197),and sequencing confirmed that the amplified products were in-deed Hh 16S rRNA gene fragments.Hh 16S rRNAgene presence was detected in patients with all five diseases,and the highest detec-tion rate was observed in patients with colorectal cancer,at 11.1%(15/135).Among 149 healthy volunteers,the detection rate of HpSA was 11.4%(17/149),only one case tested positive for the Hp ureA gene,and the Hh 16S rRNA gene was undetectable in all samples.In conclusion,Hh infection was detected in patients with digestive tract diseases.Beyond patients with gastric cancer,the prevalence of Hp infection was also notably high among patients with colorectal cancer,liver cancer,and cholangiocarcinoma.Further investigation is warranted to elucidate the roles of the two species of Helicobacter in the occurrence and progression of digestive tract cancers.

This study was aimed at investigating the presence of Helicobacter hepaticus(Hh)infection in patients with digestive tract diseases and evaluating Helicobacter pylori(Hp)infection status in patients with digestive tract cancers other than gastric cancer.Fecal samples were collected from 197 patients with digestive tract diseases at the Affiliated Cancer Hospital of Guizhou Medical Uni-versity and from 149 healthy volunteers residing in Guiyang.Hp stool antigen(HpSA)was detected with the colloidal gold method.Af-ter the extraction of fecal DNA,the Hp specific ureA gene and the Hh specific 16S rRNA gene were amplified via nested PCR,and the amplified products were subsequently confirmed through sequencing analysis.The study included 197 patients with digestive system diseases,comprising 135 cases of colorectal cancer,32 cases of chronic gastritis,22 cases of gastric cancer,5 cases of liver cancer,and 3 cases of cholangiocarcinoma.The detection rate of HpSA was 31.5%(62/197).HpSA was detected across all five disease catego-ries,and the highest detection rate was observed in patients with gastric cancer,at 50.0%(11/22),or colorectal cancer,at 24.4%(33/135).The positivity rate of Hp ureA gene PCR was 7.6%(15/197),and sequencing confirmed that the amplified products were in-deed Hp ureA gene fragments.Notably,the highest detection rate was observed in patients with colorectal cancer,at 8.9%(12/135).The positivity rate of Hh 16S rRNA gene PCR was 11.2%(22/197),and sequencing confirmed that the amplified products were in-deed Hh 16S rRNA gene fragments.Hh 16S rRNAgene presence was detected in patients with all five diseases,and the highest detec-tion rate was observed in patients with colorectal cancer,at 11.1%(15/135).Among 149 healthy volunteers,the detection rate of HpSA was 11.4%(17/149),only one case tested positive for the Hp ureA gene,and the Hh 16S rRNA gene was undetectable in all samples.In conclusion,Hh infection was detected in patients with digestive tract diseases.Beyond patients with gastric cancer,the prevalence of Hp infection was also notably high among patients with colorectal cancer,liver cancer,and cholangiocarcinoma.Further investigation is warranted to elucidate the roles of the two species of Helicobacter in the occurrence and progression of digestive tract cancers.

Objective:This study aimed to clarify the clinical characteristics of children infected with rhinovirus in the context of the Corona Virus Disease 2019 (COVID-19) pandemic and to explore the impact of recent COVID-19 infection history on their clinical features.Methods:Clinical data and laboratory test result of 286 children diagnosed with rhinovirus infection at Hangzhou Children′s Hospital from July 2022 to October 2023 were collected. A retrospective survey was conducted to determine whether all study participants had a history of COVID-19 infection within the 6 months prior to hospitalization.Results:Among the 286 children with rhinovirus infection, 180 (62.94%) had simple rhinovirus infection, while 106 (37.06%) had co-infections with other pathogens; Among the 180 rhinovirus simplex-positive children, 56.67% had wheezing symptoms; among them, 15 cases (15/180, 8.33%) were diagnosed with acute asthma attacks; 7 cases (7/180, 3.88%) were diagnosed with severe pneumonia. Based on whether the children had a history of COVID-19 infection in the 6 months prior to hospitalization, they were divided into a group with previous COVID-19 infection and a group without previous COVID-19 infection. There were no significant differences between the two groups in terms of gender, age of onset, peak fever, incidence of wheezing, incidence of pneumonia, proportion of severe pneumonia, proportion of severe asthma attacks, duration of fever, time to relief of wheezing, length of stay, white blood cell count, eosinophil count, C-reactive protein, procalcitonin, immunoglobulin E, oxygen therapy requirements, and use of intravenous steroids ( P>0.05). Conclusions:A history of COVID-19 infection in the past 6 months does not exacerbate the clinical symptoms of children with rhinovirus infection, nor does it increase the incidence of wheezing.

Objective:To investigate the positive rate and isolate Bordetella pertussis in children with suspected whooping cough and their close family members in Zhejiang Province, and further explore the susceptibility and resistant mechanism of Bordetella pertussis to antibiotics. Methods:A total of 273 nasopharynx swabs specimens from children with suspected whooping cough in Hangzhou Children′s Hospital from May 2022 to October 2022 were collected. The strains were isolated and cultured using charcoal select agar plate. Pertussis target genes were detected by RT-PCR. E-test method was used to detect the sensitivity of Bordetella pertussis strains to different antibiotics. The mechanism of resistance of Bordetella pertussis to macrolides was analyzed by whole genome sequencing. The phylogenetic analysis of isolated strains was based on core genome multilocus sequence typing(cgMLST). Results:Among 273 clinical samples of children with suspected pertussis and their close family members, 168 samples were positive by fluorescence quantitative PCR, accounting for 61.54%, and 30 pertussis strains were successfully isolated with a positive rate of 10.98%. In addition, among the 143 samples of close family members, 54.55% (78/143) samples were positive by RT-PCR and 9.79% (14/143) samples were positive by culture, suggesting that the close family member are important in family transmission of pertussis. Besides, most of the positive samples were from mothers. The results of E-test showed that 96.67%(29/30) strains showed high resistance to azithromycin with MIC value>256 mg/L, and the resistant mechanism of azithromycin was A2047G mutation in 23S rRNA. The phylogenetic analysis based on the cgMSLT showed that the isolated strains were clustered into two new different clades.Conclusions:The positive rate of Bordetella pertussis in close family members is at a high level and the mother may be the main source of infection, which is of great significance for monitoring and laboratory detection of suspected children′s family members. Bordetella pertussis shows high resistance to macrolides in Zhejiang Province, so monitoring of the antimicrobial resistance should be further strengthened to provide theoretical basis for clinical treatment and drug guidance.

Objective:Based on the Delphi method, the electronic intelligent kanban system for general ward nursing was constructed, and its clinical application was studied.Methods:This study was the quasi experimental research, 39 nursing staff working in the experimental ward of electronic intelligent kanban in general surgery of Zhongshan Hospital Fudan University in November 2021 and November 2022 were selected, Delphi expert consultation was used to develop the menus and detailed contents of general ward nursing electronic intelligent kanban system, and it was used in clinical practice. We used the Chinese version of the Nursing Assessment of Shift Report to evaluate the effect of nursing staff before and after the application.Results:The authority coefficients of the two rounds of expert consultation were 0.868 and 0.886 respectively, and the Kendall coordination coefficients of expert consultation were 0.068 and 0.076 respectively (all P<0.01). Finally, the electronic intelligent kanban consisted of 4 first-level menus, 8 second-level menus and 13 third-level menus. After the application of electronic intelligent kanban, the score of Nursing Assessment of Shift Report increased from (79.62 ± 7.64) to (83.51 ± 2.47), with a statistically significant difference ( t=-3.03, P<0.05). Conclusions:The constructed nursing electronic intelligent kanban system was scientific and reliable, which will be beneficial to improve the the effect of nurse shift handover.

Flagella are the main motility structure of Clostridioides difficile that affects the adhesion, colonization, and virulence of C. difficile in the human gastrointestinal tract. The FliL protein is a single transmembrane protein bound to the flagellar matrix. This study aimed to investigate the effect of the FliL encoding gene flagellar basal body-associated FliL family protein (fliL) on the phenotype of C. difficile. The fliL gene deletion mutant (ΔfliL) and its corresponding complementary strains (: : fliL) were constructed using allele-coupled exchange (ACE) and the standard molecular clone method. The differences in physiological properties such as growth profile, antibiotic sensitivity, pH resistance, motility, and spore production ability between the mutant and wild-type strains (CD630) were investigated. The ΔfliL mutant and the : : fliL complementary strain were successfully constructed. After comparing the phenotypes of strains CD630, ΔfliL, and : : fliL, the results showed that the growth rate and maximum biomass of ΔfliL mutant decreased than that of CD630. The ΔfliL mutant showed increased sensitivity to amoxicillin, ampicillin, and norfloxacin. Its sensitivity to kanamycin and tetracycline antibiotics decreased, and the antibiotic sensitivity partially returned to the level of CD630 strain in the : : fliL strain. Moreover, the motility was significantly reduced in the ΔfliL mutant. Interestingly, the motility of the : : fliL strain significantly increased even when compared to that of the CD630 strain. Furthermore, the pH tolerance of the ΔfliL mutant significantly increased or decreased at pH 5 or 9, respectively. Finally, the sporulation ability of ΔfliL mutant reduced considerably compared to the CD630 strain and recovered in the : : fliL strain. We conclude that the deletion of the fliL gene significantly reduced the swimming motility of C. difficile, suggesting that the fliL gene is essential for the motility of C. difficile. The fliL gene deletion significantly reduced spore production, cell growth rate, tolerance to different antibiotics, acidity, and alkalinity environments of C. difficile. These physiological characteristics are closely related to the survival advantage in the host intestine, which is correlated with its pathogenicity. Thus, we suggested that the function of the fliL gene is closely related to its motility, colonization, environmental tolerance, and spore production ability, which consequently affects the pathogenicity of C. difficile.
Humans ; Clostridioides/metabolism* ; Clostridioides difficile/metabolism* ; Bacterial Proteins/metabolism* ; Virulence ; Anti-Bacterial Agents/metabolism*

The functionality of DNA biomacromolecules has been widely excavated, as therapeutic drugs, carriers, and functionalized modification derivatives. In this study, we developed a series of DNA tetrahedron nanocages (Td),