Objective:To establish a digital reverse transcription polymerase chain reaction（dRT-PCR）detection method for hepatitis E virus（HEV）using nanoplates，and to provide technical reference for HEV monitoring in shellfish by combining virus enrichment pretreatment methods.Methods:The annealing temperature，primer and probe concentrations of HEV dRT-PCR were optimized，and the specificity of the method was evaluated；the sensitivity of this method for detecting HEV in water samples and oyster extracts was compared. The inhibition rate and recovery rate of HEV detection in artificially contaminated oyster samples were calculated，commercially available oyster samples were tested，and compare them with real-time fluorescence quantitative RT-PCR（qRT-PCR）method.Results:The optimized annealing temperature for HEV dRT-PCR was determined to be 60 ℃，and the final concentrations of primers and probes were 0.4 μmol/L，0.4 μmol/L，and 0.2 μmol/L，respectively，indicating good specificity. The sensitivity of both methods for detecting HEV RNA in water samples was higher than that in oyster extracts. The recovery rates of HEV in oyster specimens contaminated with HEV fecal suspension by dRT-PCR and qRT-PCR were 18.76% and 18.36%，respectively，with no statistically significant difference（ P>0.05）；the inhibition rates were 17.26% and 9.58%，respectively，with statistically significant differences（ P<0.05）；55 commercially available oyster samples were tested，and both methods detected HEV RNA positivity in the same sample. Conclusion:The dRT-PCR method established in this study，combined with “proteinase K digestion，PEG/NaCl precipitation，and chloroform/n-butanol extraction” pretreatment，has a good recovery effect on HEV in shellfish food containing a large amount of PCR inhibitors，and can achieve absolute quantification. It has certain application value in monitoring and risk assessment of HEV in shellfish food.

Objective:To establish a digital reverse transcription polymerase chain reaction（dRT-PCR）detection method for hepatitis E virus（HEV）using nanoplates，and to provide technical reference for HEV monitoring in shellfish by combining virus enrichment pretreatment methods.Methods:The annealing temperature，primer and probe concentrations of HEV dRT-PCR were optimized，and the specificity of the method was evaluated；the sensitivity of this method for detecting HEV in water samples and oyster extracts was compared. The inhibition rate and recovery rate of HEV detection in artificially contaminated oyster samples were calculated，commercially available oyster samples were tested，and compare them with real-time fluorescence quantitative RT-PCR（qRT-PCR）method.Results:The optimized annealing temperature for HEV dRT-PCR was determined to be 60 ℃，and the final concentrations of primers and probes were 0.4 μmol/L，0.4 μmol/L，and 0.2 μmol/L，respectively，indicating good specificity. The sensitivity of both methods for detecting HEV RNA in water samples was higher than that in oyster extracts. The recovery rates of HEV in oyster specimens contaminated with HEV fecal suspension by dRT-PCR and qRT-PCR were 18.76% and 18.36%，respectively，with no statistically significant difference（ P>0.05）；the inhibition rates were 17.26% and 9.58%，respectively，with statistically significant differences（ P<0.05）；55 commercially available oyster samples were tested，and both methods detected HEV RNA positivity in the same sample. Conclusion:The dRT-PCR method established in this study，combined with “proteinase K digestion，PEG/NaCl precipitation，and chloroform/n-butanol extraction” pretreatment，has a good recovery effect on HEV in shellfish food containing a large amount of PCR inhibitors，and can achieve absolute quantification. It has certain application value in monitoring and risk assessment of HEV in shellfish food.

Objective:To systematically and comprehensively retrieve studies on urethra care for patients with urinary catheter at home and abroad, so as to provide a reference for removing low-value care measures in the future.Methods:Arksey and O'?Malley scoping review method was used to systematically search the China National Knowledge Infrastructure, China Biology Medicine disc, WanFang Data, VIP, PubMed, Cochrane Library, CINAHL, Embase, and Web of Science. The search period was from database establishment to January 1, 2024. Literature was screened based on inclusion and exclusion criteria, and catheter care site, disinfection/cleaner type, frequency, institutional norms, and determinants were independently extracted by two researchers, where determinants were framed based on the theoretical domains framework.Results:A total of 21 papers were included. Urethra low-value care measures were reflected in the selection of nursing solutions that differed significantly from guideline recommendations and were inconsistently standardized across hospitals. Thirteen strategies were proposed to remove urethra low-value care measures. Factors that influenced the removal of low-value care behaviors included knowledge, reinforcement, environment and resources, social influences, personal roles and identities, outcome beliefs and memories, and attention and decision-making processes.Conclusions:Future researchers should further refine the evidence on urethra care and develop contextualized care for complex, multilevel healthcare settings to achieve continuous improvement and high quality care at low cost.

Atrial fibrillation is the most common type of chronic cardiac arrhythmia. Catheter ablation and left atrial appendage occlusion are effective treatment methods for atrial fibrillation， but these procedures require anesthesia support. However， anesthetic drugs often cause side effects such as nausea， vomiting， involuntary movements， and respiratory depression. This paper presents a case of a successful one-stop procedure for cryoballoon ablation and left atrial appendage occlusion of atrial fibrillation performed entirely under acupuncture anesthesia. Thirty minutes before the procedure， acupuncture needles were inserted perpendicularly at bilateral Neiguan （PC 6）， Lieque （LU 7）， Ximen （PC 4） and （LU 6）. After obtaining the deqi （得气） sensation， an electroacupuncture device was connected， and electroacupuncture anesthesia was used for pain control throughout the procedure. The patient exhibited good tolerance and cooperation， with electroacupuncture anesthesia completely replacing intravenous anesthetics， ensuring the smooth completion of the surgery. Postoperative follow-up showed favorable outcomes.

Objective:To compare enrichment and nucleic acid detection method for hepatitis E virus (HEV) in simulated cola samples.Methods:Cola samples experimentally contaminated with HEV were enriched using positively charged filter membrane-direct lysis (Method 1), tangential flow ultrafiltration membrane-direct lysis (Method 2), and Method 3 and 4 (with the addition of a PCR inhibitor removal step on the basis of Method 1 and 2, respectively), and were assayed by real-time fluorescence quantitative RT-PCR(RT-qPCR), and the recoveries and inhibition rates were compared. Digital RT-PCR(RT-dPCR) and RT-qPCR were applied to detect the recovery of HEV in different medium and low concentrations of experimentally contaminated cola samples; and the inhibition rate and sensitivity of HEV RNA detection in different matrices.Methods 3 was selected for virus enrichment of 8 commercially available cola specimens, RT-qPCR and RT-dPCR for HEV RNA detection.Results:The HEV recoveries of method 3 and 4 (10.44% and 10.16%) were higher than those of method 1 and 2 (4.89% and 0.32%), and the differences were statistically significant ( P<0.05). The inhibition rates of method 3 and 4 were smaller than the inhibition rates of method 1 and 2. The recoveries of HEV in medium concentration artificially contaminated cola samples by RT-qPCR and RT-dPCR were 17.04% and 16.28%, respectively, and for low concentration artificially contaminated cola samples were 6.91% and 4.65%, respectively, and the differences in recoveries between the two assays at the same concentration were not statistically significant ( P=0.260, P=0.107 ); Cola matrix inhibits the detection of both RT-qPCR and RT-dPCR assays. Eight commercially available cola specimens were negative for HEV. Conclusions:Detection of HEV in cola beverages can be done by positively charged filter membrane-direct lysis + inhibitor removal (method 3) or tangential flow ultrafiltration membrane-direct lysis + inhibitor removal (method 4) enrichment, followed by RT-dPCR or RT-qPCR, with a high recovery of virus detection.

Objective:To systematically and comprehensively retrieve studies on urethra care for patients with urinary catheter at home and abroad, so as to provide a reference for removing low-value care measures in the future.Methods:Arksey and O'?Malley scoping review method was used to systematically search the China National Knowledge Infrastructure, China Biology Medicine disc, WanFang Data, VIP, PubMed, Cochrane Library, CINAHL, Embase, and Web of Science. The search period was from database establishment to January 1, 2024. Literature was screened based on inclusion and exclusion criteria, and catheter care site, disinfection/cleaner type, frequency, institutional norms, and determinants were independently extracted by two researchers, where determinants were framed based on the theoretical domains framework.Results:A total of 21 papers were included. Urethra low-value care measures were reflected in the selection of nursing solutions that differed significantly from guideline recommendations and were inconsistently standardized across hospitals. Thirteen strategies were proposed to remove urethra low-value care measures. Factors that influenced the removal of low-value care behaviors included knowledge, reinforcement, environment and resources, social influences, personal roles and identities, outcome beliefs and memories, and attention and decision-making processes.Conclusions:Future researchers should further refine the evidence on urethra care and develop contextualized care for complex, multilevel healthcare settings to achieve continuous improvement and high quality care at low cost.

Objective:To compare enrichment and nucleic acid detection method for hepatitis E virus (HEV) in simulated cola samples.Methods:Cola samples experimentally contaminated with HEV were enriched using positively charged filter membrane-direct lysis (Method 1), tangential flow ultrafiltration membrane-direct lysis (Method 2), and Method 3 and 4 (with the addition of a PCR inhibitor removal step on the basis of Method 1 and 2, respectively), and were assayed by real-time fluorescence quantitative RT-PCR(RT-qPCR), and the recoveries and inhibition rates were compared. Digital RT-PCR(RT-dPCR) and RT-qPCR were applied to detect the recovery of HEV in different medium and low concentrations of experimentally contaminated cola samples; and the inhibition rate and sensitivity of HEV RNA detection in different matrices.Methods 3 was selected for virus enrichment of 8 commercially available cola specimens, RT-qPCR and RT-dPCR for HEV RNA detection.Results:The HEV recoveries of method 3 and 4 (10.44% and 10.16%) were higher than those of method 1 and 2 (4.89% and 0.32%), and the differences were statistically significant ( P<0.05). The inhibition rates of method 3 and 4 were smaller than the inhibition rates of method 1 and 2. The recoveries of HEV in medium concentration artificially contaminated cola samples by RT-qPCR and RT-dPCR were 17.04% and 16.28%, respectively, and for low concentration artificially contaminated cola samples were 6.91% and 4.65%, respectively, and the differences in recoveries between the two assays at the same concentration were not statistically significant ( P=0.260, P=0.107 ); Cola matrix inhibits the detection of both RT-qPCR and RT-dPCR assays. Eight commercially available cola specimens were negative for HEV. Conclusions:Detection of HEV in cola beverages can be done by positively charged filter membrane-direct lysis + inhibitor removal (method 3) or tangential flow ultrafiltration membrane-direct lysis + inhibitor removal (method 4) enrichment, followed by RT-dPCR or RT-qPCR, with a high recovery of virus detection.

Biological hydroxyapatite(BHA)is widely used in the treatment of clinical bone defects due to its good biocompatibility and osteoconductivity.The clinical application of mateiral size is based on the principle of bone defect area adaptation,which contributes to diversity of BHA sizes.However,different sizes correspond to different hierarchi-cal levels of bone biomimicry.As the size changes,the bone biomimicry hierarchy evolves accordingly and influences the process of bone repair and regeneration through osteo-coagulo-immunomodulation,leading to unstable bone graft outcomes.Therefore,this paper reviews the size effect of clinical BHA,analyzes the multilevel structure of natural bone,proposes the evolution of bone biomimetic hierarchy triggered by the size of BHA,and further analyzes the size-media-ted osteo-coagulo-immunomodulation.Based on the hierarchical levels of bone and its osteo-coagulo-immunomodula-tion effect,we provide a new understanding of the biolog-ical principle of the size effect of biomaterials and a theo-retical basis for the basic research and clinical application of different size BHA materials.

Objective To summarise and analyse the qualitative studies on the factors that influence patient involvement in decision-making for the initial administration of insulin for the patients with Type II diabetes,from the perspectives of patients and healthcare staff in order to provide a reference to promote patient involvement in decision-making.Methods Systematic searches were conducted across databases,such as CINAHL,Cochrane Library,EMBASE,PubMed,Web of Science,PsycINFO,China National Knowledge Infrastructure(CNKI),Wanfang Data,VIP,and SinoMed,for qualitative studies on the factors that affect patient involvement in initial insulin decision-making for the patients with Type II diabetes.The search was limited to articles from the inception of the databases to 30th September,2023.Quality of the included studies was assessed using the Joanna Briggs Institute(JBI)evidence-based healthcare centre for qualitative research quality assessment tool.The results were integrated using a synthesising integration method.Results A total of 19 articles were included,yielding 20 study results,which were categorised into 7 themes of patient decision-making related values,patient role preferences in decision-making,condition of patient,the role of healthcare staff in patient participation in decision-making,professional quality of healthcare staff,relationship between patient and healthcare staff,and the support from a medical institution.The data were ultimately integrated into 4 overarching themes of patient personal factors,healthcare staff factors,patient-staff interaction factors and medical institution factors.Conclusion The involvement of the patients with Type II diabetes in the decision-making for the initial administration of insulin is influenced by patients themselves,healthcare staff and medical institutions.It requires efforts of multiple parties:not only with the patients actively participate in decision-making,but also with the healthcare staff and institutions who provide effective decision supports.

Objective:To compare the detection method of hepatitis E virus (HEV) in simulated water samples, and to provide a reference for the detection of HEV in water.Methods:HEV fecal suspension was added to tap water or distilled water simulated water samples, and pretreatment was carried out by electropositive filter-organic eluent elution method (Method 1) to compare the extraction effect of the three nucleic acid extraction kits, A, B, and C. The simulated water samples were pre-treated by Method 1, 2 (electropositive filter-direct lysis method), 3 (tangential-flow ultrafiltration membrane-organic eluent elution method), and 4 (tangential-flow ultrafiltration membrane-direct lysis method) for pretreatment, A kit for nucleic acid extraction, Real time RT-PCR method for detection and comparison of the recovery rate; comparison of the recovery rate of different concentrations of HEV in simulated water samples; comparing the inhibitory effects of inhibitors in tap water samples on real time RT-PCR; and detection of HEV in different batches of tap water specimens.Results:Kit A nucleic acid extraction was better; the recoveries of method 1, 2, 3 and 4 were 7.31%, 39.88%, 6.85% and 64.88%, respectively, which showed a statistically significant difference in the recoveries ( F=114.069, P<0.001). The recoveries of method 4 with the addition of high, medium and low concentrations of HEV were 65.26%, 42.76% and 32.79%, respectively. The inhibition of all four pre-treatment method was less than 75%, which meets the requirements of ISO (15216-2∶2019). Twenty tap water specimens were tested for HEV and the result were negative. Conclusions:This study showed that the two membranes better recovered in combination with direct lysis, respectively; Methods 4 had a higher recovery in the detection of HEV in small volumes of distilled or tap water, but it was limited by the volume of water samples, turbidity, and so on. Suitable method can be selected for different water quality and laboratory conditions.