Objective:To identify candidate strains of Nontoxigenic Clostridioides difficile (NTCD) with potential for intervention in Clostridioides difficile infection (CDI) and analyze their phenotypic characteristics. Methods:A total of 713 Clostridioides difficile strains from various sources were systematically collected nationwide between 2015 and 2023. This included 649 strains isolated from human fecal samples and 64 strains isolated from the fecal samples of farmed animals. NTCD strains were preliminarily screened through toxin gene detection and antibiotic sensitivity test, and then NTCD candidate strains with potential for intervention in CDI were screened by a series of in vitro experiments, including MLST, sporulation, germination, adhesion, motility, and biofilm formation ability. Ultimately, the virulence genes and antimicrobial resistance genes of the candidate strains were comprehensively analyzed to rigorously assess their safety profiles. Results:Among 713 strains of C. difficile from different sources, 10 strains were initially screened out, which were non-toxin-producing and sensitive to antibiotics. MLST showed that seven strains were from the Clade1 branch and three strains were of a novel type. The results of sporulation and germination showed that SD59, SD178, SJZ17, and WZ142 had stronger sporulation and germination abilities. The adhesion of 10 strains was high, and the adhesion rate was between 72.93% and 99.32%. The motility of all strains was different, and the motility of SD178, SD59 and SJZ17 was stronger. The biofilm-forming ability of all strains was weak. SD59, SD178 and SJZ17 carried a limited number of virulence and resistance genes, thereby posing a relatively low safety risk. Conclusion:Three NTCD strains are successfully selected as potential effective NTCD strains to interfere with CDI.

Objective:To identify candidate strains of Nontoxigenic Clostridioides difficile (NTCD) with potential for intervention in Clostridioides difficile infection (CDI) and analyze their phenotypic characteristics. Methods:A total of 713 Clostridioides difficile strains from various sources were systematically collected nationwide between 2015 and 2023. This included 649 strains isolated from human fecal samples and 64 strains isolated from the fecal samples of farmed animals. NTCD strains were preliminarily screened through toxin gene detection and antibiotic sensitivity test, and then NTCD candidate strains with potential for intervention in CDI were screened by a series of in vitro experiments, including MLST, sporulation, germination, adhesion, motility, and biofilm formation ability. Ultimately, the virulence genes and antimicrobial resistance genes of the candidate strains were comprehensively analyzed to rigorously assess their safety profiles. Results:Among 713 strains of C. difficile from different sources, 10 strains were initially screened out, which were non-toxin-producing and sensitive to antibiotics. MLST showed that seven strains were from the Clade1 branch and three strains were of a novel type. The results of sporulation and germination showed that SD59, SD178, SJZ17, and WZ142 had stronger sporulation and germination abilities. The adhesion of 10 strains was high, and the adhesion rate was between 72.93% and 99.32%. The motility of all strains was different, and the motility of SD178, SD59 and SJZ17 was stronger. The biofilm-forming ability of all strains was weak. SD59, SD178 and SJZ17 carried a limited number of virulence and resistance genes, thereby posing a relatively low safety risk. Conclusion:Three NTCD strains are successfully selected as potential effective NTCD strains to interfere with CDI.

OBJECTIVE To investigate the efficacy and safety of Wuling capsules combined with low-dose quetiapine in the treatment of oldest-old patients with insomnia accompanied by anxiety and depression. METHODS The clinical data of 96 oldest- old patients （aged≥80 years） with insomnia accompanied by anxiety and depression who were attending outpatient clinics or hospitalized in our hospital from June 2020 to December 2022 were retrospectively analyzed. According to the different drug treatments， the patients were divided into Wuling capsules group （0.99 g， tid， 34 cases）， quetiapine group （25-50 mg， qn， 30 cases） and combination group （using Wuling capsules and quetiapine simultaneously， same as the single drug groups， 32 cases）. Before and after 8 weeks of treatment， the patients of three groups were compared in terms of photoplethysmography sleep monitoring indexes （total sleep duration， sleep efficiency， sleep latency， and the number of awakening）， Pittsburgh sleep quality index （PSQI） score， 14-item Hamilton anxiety scale （HAMA-14） score， 17-item Hamilton depression scale （HAMD-17） score， mini-mental state examination （MMSE） score， and serum levels of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α）； the therapeutic effects and adverse drug reactions of the three groups were observed and compared. RESULTS Compared with before treatment， the total sleep time， sleep efficiency and MMSE scores of the three groups were significantly longer or higher after treatment （P＜0.05）， while the sleep latency， number of awakening， PSQI score， HAMA-14 score， HAMD-17 score， serum IL-1β and TNF-α levels were significantly shorter or lower after treatment （P＜0.05）. Sleep latency， number of awakening， HAMA-14 score， HAMD-17 score and serum TNF-α level in quetiapine group were significantly shorter or lower than Wuling capsules group after treatment （P＜0.05）. Moreover， the above indexes in the combination group were significantly better than quetiapine group and Wuling capsules group （P＜0.05）. The total effective rate of treatment in the combination 1731786794@qq.com group was significantly higher than the other two groups （P＜0.05）. There was no significant difference in the incidence of mail：2408797210@qq.com adverse drug reactions among the three groups （P＞0.05）. CONCLUSIONS The therapeutic effect of Wuling capsules combined with low-dose quetiapine is better than that of individual drugs for oldest-old patients with insomnia accompanied with anxiety and depression， not increasing the risk of adverse reactions.

Objective:To understand molecular characteristics and antibiotic resistance of Clostridioides ( C.) difficile isolated from children in China, and provide data support the development of disease risk assessment and burden studies. Methods:A total of 155 strains of C. difficile isolated from children aged <12 years in 14 provinces (autonomous regions, municipalities) in China from 2010 to 2023 were used for the analyses on molecular characteristics and antibiotic resistance of C. difficile by PCR and drug susceptibility test. Results:A total of 26 sequence types (STs) and 18 ribotypes (RTs) were identified in the 155 C. difficile isolates, in which ST3 (20.65%), ST54 (16.13%), ST35 (12.90%), and RT012/ICDC007 (14.84%), RT001/ICDC001 (11.61%), RT046/ICDC018 (8.39%) were the most common. One highly virulent strain with RT078 and 27 non-toxin-producing strains were also found; the predominant toxin gene was tcdA+ tcdB+ cdt-. All the strains were sensitive to metronidazole and vancomycin, and there were 29 multidrug-resistant strains, in which 1 strain was resistant to all the seven antibiotics except for vancomycin and metronidazole. Conclusions:Molecular characteristics and antibiotic resistance of C. difficile in children were similar to those in whole population in China, but there were regional distribution differences. It is necessary to strengthen the routine drug-resistance surveillance for C. difficile infection in children in China.

Objective:To investigate the molecular epidemiological characteristics and antibiotic resistance of Clostridioides difficile ( Cd) in hospitalized diarrhea patients in a tertiary hospital in Shaanxi Province. Methods:This study collected 425 stool samples of hospitalized diarrhea patients from October 2018 to December 2021 for isolation and identification of Cd. Toxin genes carried by the isolates were detected. Multilocus sequence typing (MLST) was performed to analyze the phylogenetic profile. Antibiotic susceptibility was analyzed by E-test. Results:Forty-nine strains of Cd were isolated from the 425 samples, including 37 strains of toxigenic Cd (75.5%, 37/49). The detection rate of Cd was 14.0% (25/179) in diarrhea patients aged ≥65 years old and 36.4% (4/11) in Nephrology Department. In the 37 toxigenic Cd strains, A -B + CDT -Cd, A + B + CDT -Cd and A + B + CDT +Cd accounted for 18.9% (7/37), 78.4% (29/37) and 2.7% (1/37), respectively. There were 24 ST types, among which ST2, ST3 and ST54 were the predominant types, each accounting for 12.2% (6/49). All strains were sensitive to metronidazole and vancomycin, with a high resistance rate of 93.9% (46/49) to ciprofloxacin and a low resistance rate of 12.2% (6/49) and 10.2% (5/49) to rifampicin and meropenem, respectively. Conclusions:The main type of toxigenic strains was A + B + CDT -. ST2, ST3 and ST54 were the predominant types and the distribution of ST types was scattered. All isolates were sensitive to metronidazole and vancomycin and most of them were resistant to ciprofloxacin.

Objective: To establish a standard operation procedure (SOP) for ribosome genotyping (ribotyping) on Clostridioides (C.) difficile, supplement and verify ribotyping typing library, so as to improve the comparability of data between different laboratories and to develop surveillance network of C. difficil in China. Methods: Molecular typing of 54 reference strains from the United States and Europe of C. difficile were performed by using the SOP referencing correspondence from abroad and from our laboratory with a BioNumerics 7.6 software to estimate the reference library of types of C. difficile. Identification of 374 clinical and animal isolates of C. difficile from 13 cities in China between 2010 and 2018, to supplement the library information. Kappa test was used to evaluate the consistency. Results: Results of capillary electrophoresis of reference strains appeared clear and stable, which guaranteed the clustering results being fast and accurate. Results from the supplementary typing showed that there were 84 types of isolates, of which 25 RT types were consistent with reference strains from abroad, while 58 RT types were different from referenced types. In the 40 referenced types, 15 RT types were not found in this study. In the consistency evaluation, the Kappa value was 0.891 and (P<0.01), showing the two Molecular typing as consistent and with close resemblance. Conclusions The result of capillary electrophoresis by applying SOP for ribotyping on C. difficile base on QIAxcel capillary electrophoresis system, appeared clear and stable. The standardized library seemed more easily used for comparability and data sharing between the laboratories.

Objective To develop a shoe-cover-style obstacle avoidance device based on gait detection algorithm for the visually impaired.Methods The device was composed of shoes covers and an electric control box.The hardware involved in MSP430F1132 SCM as the control unit,triaxial accelerometer to measure foot acceleration,ultrasonic sensor to detect obstacles and solar battery for power supply.A gait detection algorithm was proposed to implement a foot motion states detection program,and the motion states included static and moving ones.Results The device executed obstacle detection when'the feet were raised and let down,and obstacle detection were stopped in case the feet were kept unmoving,shaking and hanging.The rate for mis-alarm was decreased and the operation procedure was simplified.Conclusion The device behaves well in timeliness,efficiency,mis-alarm,portability,low cost and easy operation,and thus is worthy studying and practical application.

AIM: To investigate the effect of venous marker chicken ovalbumin upstream promoter-transcription factor Ⅱ (COUP-TFⅡ) expression on vascular endothelial cell senescence and its molecular mechanism.METHODS: The mRNA expression of COUP-TFⅡ in the human coronary artery endothelial cells (HCAEC) and human umbilical vein endothelial cells (HUVEC) was detected by RT-qPCR.After transfection with a COUP-TFⅡ siRNA (siCOUP-TFⅡ) to inhibit COUP-TFⅡ expression in the HUVEC, the senescence and proliferation of endothelial cells were evaluated by β-galactosidase staining, Western blot, CCK-8 assay and cell counting after treatment with 10-5 mol/L angiotensin Ⅱ (AngⅡ).The protein levels of Akt and p-Akt were determined by Western blot.RESULTS: Compared with the HCAEC, COUP-TFⅡ was significantly highly expressed in the HUVEC.Knockdown of COUP-TFⅡ via siCOUP-TFⅡ significantly induced endothelial cell senescence and inhibited endothelial cell proliferation and p-Akt level after treatment with AngⅡ at 10-5 mol/L.Furthermore, an Akt activator SC79 at 4 mg/L partly reversed the effect of siCOUP-TFⅡ on AngⅡ-induced endothelial cell senescence and proliferation.CONCLUSION: Knockdown of COUP-TFⅡ promotes endothelial cell senescence and inhibits endothelial cell proliferation, which might be partly regulated by Akt signaling.

Objective: To investigate the differences of biochemical markers between the patients with pulmonary hypertension related to left heart disease (PH-LHD) and LHD; to explore the sensitive bio markers which may predict PH in LHD patients. Methods: A total of 355 LHD patients admitted to our hospital from 2014-01 to 2015-05 were enrolled. According to 2009 ESC/ERS guidelines, PH was deifned by pulmonary artery systolic pressure (PASP)>50 mmHg and patients were divided into 2 groups: LHD group,n=224 and PH-LHD group,n=131. The basic information with blood levels of biomarkers was recorded and their accuracy for predicting PH was analyzed. Results: The pathogenesis of LHD included 184 (51.83%) patients of coronary heart disease, 90 (25.35%) of dilated cardiomyopathy and 81 (22.81%) of cardiac valve heart disease. Compared with LHD group, PH-LHD group had increased ratio of NYHA III and IV degree (89.31% vs 45.54%), decreased LVEF [42.0 (33.0, 59.0) % vs 60.0 (42.0, 65.0) %], all P<0.001; PH-LHD group presented elevated blood levels of BNP, bilirubin, red cell distribution width (RDW), uric acid and cystatin C, while reduced lipoprotein (HDL), allP<0.001. PASP was positively related to biomarkers as BNP, bilirubin, RDW, uric acid and cystatin C, while negatively related to HDL. With the combination of BNP, direct bilirubin and RDW, the predictive value for PH-LHD under ROC curve was 0.828 with the sensitivity at 0.813, speciifcity at 0.708. Conclusion: Blood levels of biochemical markers were statistically different between the patients of PH-LHD and LHD; the combination of BNP, direct bilirubin and RDW showed the higher accuracy for predicting PH occurrence in LHD patients.

OBJECTIVETo study clinicopathologic features, immunohistochemical profile, diagnosis and differential diagnosis of childhood central nervous system primitive neuroectodermal tumors (CNS PNETs) with the features of ependymoblastoma and neuroblastoma.

METHODSThe clinical data, morphologic and immunohistochemical features were analyzed in 4 cases of pediatric CNS PNETs with features of ependymoblastoma and neuroblastoma. EnVision method immunohistochemistry was applied.

RESULTSFour patients including three boys and one girl presented at the age from 12 month to 4 years and three tumors located in cerebrum, one in brain stem. All tumors showed typical combined histological patterns of ependymoblastoma and neuroblastoma, demonstrating zones of true rosettes, occasional pseudovascular rosettes, and undifferentiated neuroepithelial cells in a prominent background of mature neuropils. There was focal expression of glial fibrillary acidic protein (GFAP) consistent with glial differentiation and epithelial membrane antigen (EMA) consistent with ependymal differentiation. Necrosis was seen in three cases and calcification was present in one case. Immunohistochemically, the rosettes and undifferentiated neuroepithelial cells were positive for vimentin, partially positive for GFAP and EMA but negative for synaptophysin. The tumor cells were also positive for synaptophysin in neuropils. The Ki-67 label index ranged from 20% to 60%.

CONCLUSIONSCNS PNETs with the features of ependymoblastoma and neuroblastoma is a rare tumor with poor prognosis. The tumor primarily occurs in childhood, especially infant and belongs to the family of embryonal tumors of the CNS. The morphologic, immunohistochemical and genetic features are important in differential diagnosis from other tumors of the CNS.

Antigens, Neoplasm ; metabolism ; Central Nervous System ; pathology ; Child ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Immunohistochemistry ; Infant ; Male ; Mucin-1 ; metabolism ; Neuroblastoma ; diagnosis ; pathology ; Neuroectodermal Tumors, Primitive ; diagnosis ; pathology ; Neuroectodermal Tumors, Primitive, Peripheral ; diagnosis ; pathology ; Synaptophysin ; metabolism ; Vimentin ; metabolism