Background::Statins are the first line of treatment for dyslipidemia, but their side effects often reduce medication compliance. Natto and red yeast rice are natural ingredients with lipid-lowering effects. However, the efficacy of Natto Red Yeast Rice (NRYR) supplement in combination with statins in regulating blood lipid levels has not been fully evaluated.Methods::A multicenter, double-blinded, randomized-controlled trial was conducted among individuals with low-density lipoprotein cholesterol (LDL-C) of 3.4 to 5.0 mmol/L at six sites in China, of those at moderate risk of cardiovascular disease (CVD) are prioritized. Participants are enrolled and randomly assigned into four groups by a combination of NRYR (or its placebo) and Simvastatin (or its placebo) in a ratio of 1:1:1:1. After examination at baseline, all participants underwent intervention for 3 months and two follow-up visits at 1 month and 3 months after the intervention. The primary outcome is the change in LDL-C level at 3 months, and secondary outcomes include changes in levels of other lipid profiles and biomarkers, as well as calculated 10-year CVD risk. A total of 1136 participants were randomly assigned, of whom 1110 received the intervention.Discussion::This study may provide new evidence for the efficacy of NRYR supplement in combination with statins to regulate lipid levels and optimize lipid management.Trial Registration::Chinese Clinical Trial Registry database: registration nos. ChiCTR2200064214, ChiCTR2200064215.

Background::Statins are the first line of treatment for dyslipidemia, but their side effects often reduce medication compliance. Natto and red yeast rice are natural ingredients with lipid-lowering effects. However, the efficacy of Natto Red Yeast Rice (NRYR) supplement in combination with statins in regulating blood lipid levels has not been fully evaluated.Methods::A multicenter, double-blinded, randomized-controlled trial was conducted among individuals with low-density lipoprotein cholesterol (LDL-C) of 3.4 to 5.0 mmol/L at six sites in China, of those at moderate risk of cardiovascular disease (CVD) are prioritized. Participants are enrolled and randomly assigned into four groups by a combination of NRYR (or its placebo) and Simvastatin (or its placebo) in a ratio of 1:1:1:1. After examination at baseline, all participants underwent intervention for 3 months and two follow-up visits at 1 month and 3 months after the intervention. The primary outcome is the change in LDL-C level at 3 months, and secondary outcomes include changes in levels of other lipid profiles and biomarkers, as well as calculated 10-year CVD risk. A total of 1136 participants were randomly assigned, of whom 1110 received the intervention.Discussion::This study may provide new evidence for the efficacy of NRYR supplement in combination with statins to regulate lipid levels and optimize lipid management.Trial Registration::Chinese Clinical Trial Registry database: registration nos. ChiCTR2200064214, ChiCTR2200064215.

Objective:To develop and validate the Pain nursing Competency Evaluation Questionnaire for Nursing Students to provide an effective tool for measuring the pain management competency of nursing students in China.Methods:The questionnaire was constructed through literature review, semi-structured interviews, focus group discussions, Delphi expert consultation, and a pre-survey. From September 2023 to January 2024, a convenience sampling method was used to select 250 nursing students from Hangzhou Normal University and Lishui University in Zhejiang Province for the survey. Reliability and validity of the developed questionnaire were tested. A random sample of 30 nursing students was selected for retesting after two weeks.Results:A total of 10 female experts were consulted through correspondence. The Pain Care Competency Evaluation Questionnaire for nursing students consists of 36 items. Through exploratory factor analysis, five common factors were extracted: pain health education, comprehensive pain assessment, pain screening and assessment, analgesic interventions, and analgesic side effects nursing, which together explained 61.695% of the variance. The content validity of the questionnaire was 0.96, and the item-level content validity index ranged from 0.900 to 1.000. The overall Cronbach′s α coefficient was 0.924, and the Cronbach′s α coefficients for the five dimensions ranged from 0.856 to 0.915. The test-retest reliability was 0.831. Conclusions:The Pain Care Competency Evaluation Questionnaire for nursing students developed in this study has good reliability and validity. It can be used as a tool to assess nursing students′ competency in pain care and provides a reference for the design and optimization of pain care courses and clinical practice programs for nursing students in undergraduate institutions.

Objective:To develop and validate the Pain nursing Competency Evaluation Questionnaire for Nursing Students to provide an effective tool for measuring the pain management competency of nursing students in China.Methods:The questionnaire was constructed through literature review, semi-structured interviews, focus group discussions, Delphi expert consultation, and a pre-survey. From September 2023 to January 2024, a convenience sampling method was used to select 250 nursing students from Hangzhou Normal University and Lishui University in Zhejiang Province for the survey. Reliability and validity of the developed questionnaire were tested. A random sample of 30 nursing students was selected for retesting after two weeks.Results:A total of 10 female experts were consulted through correspondence. The Pain Care Competency Evaluation Questionnaire for nursing students consists of 36 items. Through exploratory factor analysis, five common factors were extracted: pain health education, comprehensive pain assessment, pain screening and assessment, analgesic interventions, and analgesic side effects nursing, which together explained 61.695% of the variance. The content validity of the questionnaire was 0.96, and the item-level content validity index ranged from 0.900 to 1.000. The overall Cronbach′s α coefficient was 0.924, and the Cronbach′s α coefficients for the five dimensions ranged from 0.856 to 0.915. The test-retest reliability was 0.831. Conclusions:The Pain Care Competency Evaluation Questionnaire for nursing students developed in this study has good reliability and validity. It can be used as a tool to assess nursing students′ competency in pain care and provides a reference for the design and optimization of pain care courses and clinical practice programs for nursing students in undergraduate institutions.

Objective:To explore mechanism of regulation of proliferation,migration,invasion and expressions of immune factors of ovarian cancer SKOV3 cells by circSKA3/miR-3163/disintegrin-metalloproteinase 9(ADAM9)gene axis.Methods:qRT-PCR was used to detect circSKA3 and miR-3163 expressions in ovarian cancer tissues and paracancerous tissues.Western blot was used to detect ADAM9 protein level in tissues.SKOV3 cells were selected for cell experiment in vitro,and divided into sh-NC group,sh-circSKA3 group,miR-NC group,miR-3163 group,sh-circSKA3+anti-miR-3163 group,sh-circSKA3+pcDNA-ADAM9 group.circSKA3 and miR-3163 expressions in SKOV3 cells were detected by qRT-PCR.CCK-8,scratch test and Transwell were used to detect cell proliferation activity,migration and invasion ability.ELISA was used to detect expressions of TNF-α,IL-6 and IL-10.Double luciferase report experiment verified targeting relationship between circSKA3 with miR-3163,miR-3163 and ADAM9.Western blot was used to detect levels of ADAM9,matrix metalloproteinase-2(MMP-2)and MMP-9 in SKOV3 cells.Results:Com-pared with paracancerous tissues,circ-SKA3 and ADAM9 expressions were increased,and miR-3163 expression was decreased in ovarian cancer tissues(P<0.05).Compared with sh-NC group/miR-NC group,sh-circSKA3 group/miR-3163 group showed decreased cell proliferative activity and healing rate,decreased number of invasive cells,MMP-2,MMP-9 proteins and TNF-α and IL-6 expres-sions were decreased,expression of IL-10 was increased(P<0.05).circSKA3 could targeting regulate expression of miR-3163,and miR-3163 could targeting bind to ADAM9(P<0.05).Compared with sh-circSKA3 group,cell proliferation activity and scratch healing rate in sh-circSKA3+anti-miR-3163 and sh-circSKA3+pcDNA-ADAM9 groups were increased,number of invasive cells was in-creased,MMP-2,MMP-9 protein and TNF-α,IL-6 expressions were increased,while expression of IL-10 was decreased(P<0.05).Conclusion:Silencing circSKA3 expression can reduce expression of ADAM9 by up-regulating miR-3163 expression,inhibit ovarian cancer cell proliferation,migration and invasion,and regulate immune factor expression.

Objective:To explore the mechanism of Ion peptidase 1,mitochondrial(LONP1)in the progression of castration-resistant prostate cancer(CRPC). Methods:The expression levels of LONP1,N-Myc downstream-regulated gene 1(NDRG1)and androgen receptor(AR)proteins in benign prostatic hyperplasia cell line BPH-1,androgen-dependent human prostate cancer cell line LNCaP and castration-resistant prostate cancer(CRPC)cell line PC3 were examined by Western blotting.Recombinant vector carrying full-length LONP1 gene was delivered into LNCaP cells via lentiviral infection to establish stable LONP1-overexpressing(OE-LONP1)cell line,and LNCaP cells transfected with empty vector was used as the corresponding negative control(OE-NC).shRNA specifically targeting LONP1 gene(shLONP1)was delivered into PC3 cells to establish stable LONP1-silencing cell line,and PC3 cells transfected with empty shRNA vector(shNC)was used as the negative control.The effect of LONP1-overexpression and silencing on cell proliferation and invasion was analyzed by CCK-8 assay,colony formation assay and Transwell invasion assay in OE-LONP1 and shLONP1 cell lines.The effect of LONP1 on the AR/DNRG1 signaling axis was analyzed by co-immunoprecipitation assay and chromatin immunoprecipitation assay.The effect of NDRG1-silencing on the proliferation and invasion of shLONP1-expressing PC3 cells was evaluated by CCk-8 assay and Transwell assay.Tumor xenograft model was established by subcutaneously inoculating shLONP1-expressing PC3 cells and the negative control cells(PC3-shNC cells)into BALB/c nude mice.The mice were treated with DMSO or AR antagonist enzalutamide(ENZ).Then,the expression level of Ki-67 in tumor tissues was examined by immunohistochemical staining,and the cell apoptosis in tumor tissues was evaluated by TUNEL assay. Results:Compared with BPH-1 cells,LNCaP cells had lower(P＜0.05)while PC3 cells had higher(P＜0.05)expression level of LONP1 protein.The expression of AR and NDRG1 were inhibited in LNCaP cells when LONP1 was overexpressed(P＜0.05),whereas the expression of AR and NDRG1 were increased in PC3 cells when LONP1 was silenced(P＜0.05).The proliferation and invasion of LNCaP cells were promoted when LONP1 was overexpressed(P＜0.05),whereas the proliferation and invasion of PC3 cells were suppressed when LONP1 was knocked-down(P＜0.05).LONP1 can directly bind with AR to recognize the androgen response element(ARE)of NDRG1,which further inhibits the AR/NRDG1 signaling pathway to promote the progression of prostate cancer.The treatment of xenograft tumors in mouse models showed that both LONP1-silencing and ENZ application could inhibit tumor growth,and the best inhibitory effect was observed in mice treated with LONP1-silencing in combination with ENZ.The results of immunohistochemical staining and TUNEL assay indicated that the tumor tissues in the shLONP1+ENZ group had lower level of Ki-67 expression and higher level of cell apoptosis(P＜0.001). Conclusion:LONP1 is highly expressed in CRPC cell line PC3,and promotes prostate cancer progression by inhibiting AR/NDRG1 signaling transduction.

ObjectiveTo introduce the Hr gene of spontaneously mutated SHJH^hr mice into BALB/cAShjh inbred mice with clear genetic background,and provide a basis for study on the molecular mechanism of Hr gene mutation-induced abnormal phenotype and the application of this model.Methods Using a backcross-intercross breeding method guided by phenotypic monitoring, mutant genes from SHJH^hr mice bred by spontaneous mutation were introduced into inbred BALB/cAShjh mice by homozygous mutation introgression, and the mice were bred into BALB/cA.Cg.SHJH^hr (abbreviated as C.Cg.SHJH^hr) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJH^hr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJH^hr mice according to the method of GB/T 14927.1-2008. Finally, whole genome exon sequencing was utilized to detect the mutated genes in this mouse. ResultsFrom May 2018 to March 2022, a total of 10 generations of backcross-intercross were conducted to complete the construction of the C.Cg.SHJH^hr mouse line. Among the 90 SNPs loci detected, except for rs13484115 and rs13484116, all the other loci had the same genotype as the recipient mice BALB/cAShjh. The results of biochemical marker gene detection showed that all the 14 loci of the mouse were the same as those of the recipient mouse. Whole genome exon sequencing found that the mouse had 109 site mutations compared with the recipient mouse strain, including 71 synonymous mutations, 1 stopgain, 37 missense mutations, and 20 genes involved in protein sequence alterations (including the reported Hr gene). ConclusionC.Cg.SHJH^hr mice were created. Through exon sequencing and genetic analysis, three Hr mutated genes and associated mutated genes that mainly cause phenotypic variations were identified, which provides a basis for expanding the application of C.Cg.SHJH^hr mice in biomedical research.

Objective:To establish a three-dimensional spheroid expansion model of human glioma stem cells with spontaneous sphere forming and multilineage differentiation potential in vitro and investigated the contents of the proteins and lipids and their secondary components, so as to lay a foundation for further study of sphere metabolism. Methods:Human glioma stem cells GSC23 and SU3 were cultured in serum-free stem cell culture medium, respectively, and the cell spheres were harvested for about 2-3 weeks. After fixation in paraformaldehyde solution, dehydration, paraffin embedding, and sectioning, glioma-associated marker proteins were detected by immunohistochemical staining, and the protein and lipid and their secondary components contents in the spheroid tissues were analyzed by Raman imaging. One-way ANOVA was used to compare the protein, lipid and phenylalanine contents in the large sphere, medium sphere and small sphere groups.Results:Both stem cells were able to form stem cell expansion spheres resembling solid tumors within the culture dish. Immunohistochemical staining showed that the regular marker proteins of glioblastoma multiforme, CD133, nestin, epidermal growth factor receptor (EGFR), S100, Olig2, p53, Ki-67, glial fibrillary acidic protein (GFAP), vimentin, CXC chemokine receptor 4 (CXCR4), and CD34, were all expressed. Raman imaging revealed that the constructed expanded spheres of human glioma stem cells contained protein (2 930, 1 685 and 1 586 cm -1), lipid (2 845 and 1 444 cm -1), phenylalanine (1 003 cm -1) amide Ⅲ (1 250 cm -1), while there were no significant differences of protein, lipid and phenylalanine contents among the large sphere, medium sphere and small sphere groups ( P>0.05). Conclusions:We have successfully established an expanded spheroid model of human glioma stem cells in vitro, which not only exhibits the topographical characteristics and unlimited expansion ability of three-dimensional solid tumors, but also has the ability to stably store metabolically obligatory energy sources of tumor cells such as proteins and lipids, and is expected to serve as a promising tool for human glioma research in vitro.

Objective:To establish a three-dimensional spheroid expansion model of human glioma stem cells with spontaneous sphere forming and multilineage differentiation potential in vitro and investigated the contents of the proteins and lipids and their secondary components, so as to lay a foundation for further study of sphere metabolism. Methods:Human glioma stem cells GSC23 and SU3 were cultured in serum-free stem cell culture medium, respectively, and the cell spheres were harvested for about 2-3 weeks. After fixation in paraformaldehyde solution, dehydration, paraffin embedding, and sectioning, glioma-associated marker proteins were detected by immunohistochemical staining, and the protein and lipid and their secondary components contents in the spheroid tissues were analyzed by Raman imaging. One-way ANOVA was used to compare the protein, lipid and phenylalanine contents in the large sphere, medium sphere and small sphere groups.Results:Both stem cells were able to form stem cell expansion spheres resembling solid tumors within the culture dish. Immunohistochemical staining showed that the regular marker proteins of glioblastoma multiforme, CD133, nestin, epidermal growth factor receptor (EGFR), S100, Olig2, p53, Ki-67, glial fibrillary acidic protein (GFAP), vimentin, CXC chemokine receptor 4 (CXCR4), and CD34, were all expressed. Raman imaging revealed that the constructed expanded spheres of human glioma stem cells contained protein (2 930, 1 685 and 1 586 cm -1), lipid (2 845 and 1 444 cm -1), phenylalanine (1 003 cm -1) amide Ⅲ (1 250 cm -1), while there were no significant differences of protein, lipid and phenylalanine contents among the large sphere, medium sphere and small sphere groups ( P>0.05). Conclusions:We have successfully established an expanded spheroid model of human glioma stem cells in vitro, which not only exhibits the topographical characteristics and unlimited expansion ability of three-dimensional solid tumors, but also has the ability to stably store metabolically obligatory energy sources of tumor cells such as proteins and lipids, and is expected to serve as a promising tool for human glioma research in vitro.

Objective:To investigate the bacterial composition and antimicrobial resistance profile of clinical isolates from bloodstream infections in China.Methods:The clinical bacterial strains isolated from blood culture were collected during January 2020 to December 2020 in member hospitals of Blood Bacterial Resistant Investigation Collaborative System (BRICS). Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical Laboratory Standards Institute(CLSI, USA). WHONET 5.6 was used to analyze data.Results:During the study period, 10 043 bacterial strains were collected from 54 hospitals, of which 2 664 (26.5%) were Gram-positive bacteria and 7 379 (73.5%) were Gram-negative bacteria. The top 10 bacterial species were Escherichia coli (38.6%), Klebsiella pneumoniae (18.4%), Staphylococcus aureus (9.9%), coagulase-negative Staphylococci (7.5%), Pseudomonas aeruginosa (3.9%), Enterococcus faecium (3.3%), Enterobacter cloacae (2.8%), Enterococcus faecalis (2.6%), Acinetobacter baumannii (2.4%) and Klebsiella spp (1.8%). The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococcus aureus were 27.6% and 74.4%, respectively. No glycopeptide- and daptomycin-resistant Staphylococci were detected. More than 95% of Staphylococcus aureus were sensitive to rifampicin and SMZco. No vancomycin-resistant Enterococci strains were detected. Extended spectrum β-lactamase (ESBL) producing Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis were 48.4%, 23.6% and 36.1%, respectively. The prevalence rates of carbapenem-resistance in Escherichia coli and Klebsiella pneumoniae were 2.3% and 16.1%, respectively; 9.6% of carbapenem-resistant Klebsiella pneumoniae strains were resistant to ceftazidime/avibactam combination. The prevalence rate of carbapenem-resistance in Acinetobacter baumannii was 60.0%, while polymyxin and tigecycline showed good activity against Acinetobacter baumannii. The prevalence rate of carbapenem-resistance of Pseudomonas aeruginosa was 23.2%. Conclusions:The surveillance results in 2020 showed that the main pathogens of bloodstream infection in China were gram-negative bacteria, while Escherichia coli was the most common pathogen, and ESBL-producing strains declined while carbapenem-resistant Klebsiella pneumoniae kept on high level. The proportion and the prevalence of carbapenem-resistant Pseudomonas aeruginosa were on the rise slowly. On the other side, the MRSA incidence got lower in China, while the overall prevalence of vancomycin-resistant Enterococci was low.