Objective To explore the occurrence risk of enteral nutrition intolerance and analyze its influencing factors in 302 elderly critically ill patients. Methods The clinical case data of elderly critically ill patients in department of elderly cadres of the hospital were retrospectively analyzed from January 2019 to January 2024. According to the occurrence of enteral nutrition intolerance or not, they were divided into occurrence group (n=156) and non-occurrence group (n=146). The risk of nutritional intolerance in elderly critically ill patients was evaluated by feeding intolerance risk assessment form, and the influencing factors of enteral nutrition intolerance were analyzed by multivariate logistic regression analysis. Results Among the 302 elderly patients with critical illness, 53.31% (161/302) had high risk of enteral nutrition intolerance, and 51.66% (156/302) had enteral nutrition intolerance. Multivariate logistic analysis revealed that CRP level>10mg/L, APACHE-II score≥20 points, Lac≥3mmol/L and hypoalbuminemia were risk factors in elderly critically ill patients (OR=1.806, 2.977, 8.232, 3.031, P=0.011, 0.001, 0.041, 0.047), and addition of dietary fiber was a protective factor for enteral nutrition intolerance (OR=1.652, P=0.037). Conclusion The risk of enteral nutrition intolerance is high in elderly critically ill patients. Lac level, CRP level, hypoalbuminemia, and APACHE-II score of patients are independent risk factors for enteral nutrition intolerance, and addition of dietary fiber is a protective factor. It is necessary to take targeted interventions for patients according to the above factors to minimize the occurrence of enteral nutrition intolerance.

Objective To examine the impact and partial mechanism of bupivacaine sustained-release drug(code PB21)in combination with low-dose dexamethasone(Dex)on the analgesic time of rats with knee osteoarthritis(KOA).Methods Using the techniques of anterior cruciate ligament transection and meniscus instability,a rat KOA model was created.After eight weeks,SD mice were split into three groups at random:a group for the model,one for Dex(50 μg),one for PB21(1.5 mg),and one for combined administration(1.5 mg PB21/50 μg Dex),with a control group that received a sham operation.The pain thresholds of KOA rats were measured using a Pres-sure Application Measurement(PAM)at different intervals before to delivery and 4,24,36,and 48 hours following administration;to gauge changes in discomfort,a CatWalk was used to assess the rats'average foot strength and maximum contact area before,four,twenty-four,and forty-eight hours after treatment.A portion of the rats were put to sleep at four,twenty-four,and forty-eight hours following the injection,and the joint synovium was removed for paraffin sectioning.Immunohistochemistry was used to identify the expression of GAP43 in the synovium,whereas immunofluorescence was used to identify the expression of CGRP in the same tissue.Results The average strength and maximum contact area of the foot and claw decreased(P＜0.01),and the pain threshold decreased(P＜0.01)in the model group compared to the sham operation group.The PB21+Dex group experienced a delayed pain threshold lowering time delay when compared to the PB21 and Dex treatment groups alone.Up to 48 hours lat-er,the combination administration group's average strength and maximum contact area of the foot paw remained ele-vated,and there was a statistically significant difference(P＜0.05)between the combined administration group and PB21 and Dex alone.GAP43 and CGRP expression levels in synovial tissue were detected.The results indica-ted that PB21 and Dex alone could lower protein expression levels at 4 and 24 h at the two time points,and that the PB21+Dex group could still significantly lower GAP43 and CGRP expression levels at 48 h.At the 48 h time point,the PB21+Dex group was statistically significant when compared to the PB21 and Dex alone administration group(P＜0.05).Conclusion In summary low dose dexamethasone can prolong the analgesic effect of PB21 on KOA rats,which is connected to reducing the expression of pain related proteins CGRP and GAP43.

Objective:To construct a recombinant bioluminescent bacteriophage (HT7) targeting Escherichia coli, and evaluate its ability to identify Escherichia coli. Methods:Initially, pCRISPR-sg (1-10) and PFN-1000 plasmid strains were constructed by genetic engineering, and the most efficient small guild RNA (sgRNA) were screened by bilayer plate. By the gene editing technique, which comprised homologous recombination and clustered regularly interspaced short palin dromic repeats (CRISPR)-Cas system, the Nanoluc luciferase gene was integrated into the downstream non-coding region of 10A gene of T7 phage, to constructe the bioluminescent phage HT7 successfully. The difference of biological characteristics between HT7 phage and T7 phage was evaluated by plaque assay and liquid amplification assay. In addition, 51 strains of Escherichia coli, 20 strains of Klebsiella pneumoniae, 14 strains of Staphylococcus aureus, 6 strains of Enterococcus faecium, 5 strains of Enterococcus faecalis, 3 strains of Acinetobacter baumannii and 1 strain of Pseudomonas aeruginosa were collected and isolated to evaluate the limit of detection and specificity of HT7 phage. Results:Among the 10 CRISPR-targeted cleavage systems constructed, sgRNA8 exhibited the highest cleavage efficiency, with a cleavage rate of 0.18. After three rounds of recombination screening using the pCas9/pCRISPR/PFN-1000 triple-plasmid system, PCR validation yielded recombinant phage bands at 2 798 bp, indicating the successful construction of the HT7 phage. The recombinant phage showed significant differences in biological characteristics in terms of lysis efficiency ( P<0.001), one-step growth curve ( P=0.001), and infection multiplicity ( P=0.031). Both lysis burst time and log growth node were extended by 10 min, with the optimal infection multiplicity being 0.1. Clinical sample testing identified lysis of 6 strains of Escherichia coli within 4.5 h, while other strains remained unaffected, with detection of pathogenic bacteria below 10 CFU/ml. Conclusions:The developed pCas9/pCRISPR/PFN-1000 triple-plasmid editing system efficiently edits the bacteriophage genome. The constructed HT7 fluorescent bacteriophage enables the detection of Escherichia coli below 10 CFU/ml within 4.5 hours, demonstrating low detection limits and high detection specificity.

AIM:To investigate the effects of overexpressing α-Klotho(KL)in RAW264.7 cells stimulated by oxidative stress on the proliferation, migration, tube-formation and tight junction of human umbilical vein endothelial cells(HUVECs).METHODS:RAW264.7 cells were categorized into control, 4-hydroxynonenal(4HNE), and 4HNE+KL groups, with F4/80 expression assessed via immunofluorescence staining. Three groups of conditional media were prepared for HUVECs and culture divided into Mø-NC, Mø-4HNE, and Mø-4HNE+KL groups. Cell proliferation was evaluated using CCK8 assay, while scratch test and Transwell assays were employed to measure cell migration. Additionally, tube-formation assay was conducted to assess cell tubule formation, and Western blot assay was utilized to detect the protein expression levels of Claudin 5, Occludin and ZO 1.RESULTS:The results of immunofluorescence staining showed that the fluorescence intensity of F4/80 of RAW264.7 cells in the 4HNE group was significantly enhanced compared with the control group, while that of F4/80 in the 4HNE+KL group was significantly decreased compared with the 4HNE group(all P<0.05). The CCK8 assay results revealed a significant increase in the proliferation of HUVECs in the Mø-4HNE group compared with the Mø-NC group. Conversely, the proliferation of the Mø-4HNE+KL group exhibited a significant decrease compared with that in the Mø-4HNE group(all P<0.01). The results of scratch test and Transwell assays demonstrated a significant increase in the migration of HUVECs in the Mø-4HNE group compared with the Mø-NC group, while the migration of the Mø-4HNE+KL group exhibited a significant decrease compared with the Mø-4HNE group(all P<0.01). In the tube-formation assay, it was observed that the number of tubes formed by HUVECs in the Mø-4HNE group was significantly increased compared with the Mø-NC group, while that of tubes formed in the Mø-4HNE+KL group was significantly decreased compared with the Mø-4HNE group(all P<0.01). Additionally, the Western blot results revealed a significant decrease in the relative expression levels of Claudin 5, Occludin, and ZO 1 in the Mø-4HNE group compared with the Mø-NC group. Conversely, in the Mø-4HNE+KL group, there was a significant increase in the relative expression levels of Claudin 5, Occludin, and ZO 1 compared to the Mø-4HNE group(all P<0.01).CONCLUSIONS: KL inhibits the proliferation, migration, and tube-formation of HUVECs while enhancing the tight junction by changing the activation state of macrophages in the diabetic oxidative stress environment.

Objective:To investigate the characteristics of cognitive frailty and its influencing factors in older patients with chronic heart failure.Methods:In this cross-sectional study, 300 older patients hospitalized for chronic heart failure in a tertiary hospital in Shandong Province between September 2021 and September 2022 were selected.A general information questionnaire, the mini-nutritional assessment scale-short form(MNA-SF), the athens insomnia scale(AIS), the ulca loneliness scale, the geriatric depression scale-5 item version(GDS-5), and the social support rating scale(SSRS)were used for assessment and influencing factors were identified by univariate and Logistic regression analysis.Results:Among 300 older patients with chronic heart failure, the prevalence of cognitive frailty was 75.3%(226 cases). Logistic regression analysis showed that age between 70-79 years( OR=0.543, 95% CI: 0.299-0.987), education level( OR=3.644, 95% CI: 1.780-7.461), weekly intellectual activity( OR=2.168, 95% CI: 1.082-4.334)and loneliness( OR=1.101, 95% CI: 1.032-1.175)were factors influencing cognitive frailty in older patients with chronic heart failure. Conclusions:The prevalence of cognitive frailty in older patients with chronic heart failure is high, and age, education level, weekly intellectual activity and loneliness are influencing factors, with education level having the greatest impact on older patients with chronic heart failure.

Objectives:To present the epidemiological characteristics of stroke incidence and stroke-related mortality among the whole population in national cardiovascular disease surveillance areas from 2015 to 2019. Methods:Data of stroke incidence and stroke-related mortality from 2015 to 2019 were collected from the China Registry of Cardiovascular Events(China RACE),which was established in 2014,covering 100 counties(cities,districts)in 31 provinces in China.The Joinpoint model was used to analyze the annual percentage changes(APC)and trends of stroke incidence rate.The age-standardized incidence rate(ASIR)was calculated using the Seventh National Census data as the standard population.With annual reported stroke events and stroke-related deaths,the mortality to incidence ratio(M/I)were examined. Results:From 2015 to 2019,an increase of 9.41%(APC=2.12%,95%CI:1.43%-2.82%,Ptrend<0.01)resulted in the overall stroke crude incidence rate(CIR)of 468.48/100 000 in 2019 among the whole population,with relatively higher in male and in rural area.The more sharply elevating of CIR appeared in males(11.26%[APC=2.53%,95%CI:1.83%-3.24%,Ptrend<0.01])rather than in females(7.26%[APC=1.63%,95%CI:0.81%-2.46%,Ptrend<0.01]).Meanwhile,the general ASIR decreased 7.47%(APC=-1.72%,95%CI:-3.23%--0.20%,Ptrend<0.05),reaching 523.82/100 000 in 2019.The females generally showed significant descending trend(9.56%[APC=-2.27%,95%CI:-3.99%--0.52%,Ptrend<0.05]),as well as more reduction than that in the males(15.82%vs.11.40%)in urban area.The crude incidence rate of stroke increased with age.From 2015 to 2019,the CIR in 45-49 age group increased 12.48%(APC=3.18%,95%CI:1.67%-4.72%,Ptrend<0.01),compared with an reduction of 15.76%(APC=-4.39%,95%CI:-7.63%--1.04%,Ptrend<0.05)in 80-84 age group.Over the monitoring years,the overall M/I was 0.19,with an age-specific U-shaped distribution.The lowest of M/I(0.10)appeared in those aged 50-54 and 55-59,while the highest(0.45)detected in those aged 85 and over.The M/I of all age in urban areas were consistently lower than that in rural areas. Conclusions:Stroke incidence burden increased from 2015-2019 in the national surveillance areas in China,along with the unfavorable geographic diversity and age-specific divergence.Further efforts are required to improve health care covering all ages and regions in China to reduce the incidence of stroke and stroke-related mortality.

Objectives:The present study aims to investigate the incidence and mortality feature of acute myocardial infarction(AMI)from 2015 to 2019 in China by utilizing national registry data. Methods:Data of AMI incidence and mortality in the surveillance area during 2015 to 2019 were abstracted from China Registry of Acute Cardiovascular Event(China RACE),which was established in 100 counties from 31 provincial regions in China.Incidence rate,age standardized incidence rate(ASIR)and mortality to incidence ratio(M/I)was estimated in AMI cases.A Joinpoint regression was executed and annual percent change(APC)was examined to identify trends in incidence. Results:From 2015 to 2019,a total of 257 686 acute myocardial infarction incidence and 149 169 deaths were registered.The annual incidence rate of AMI in 2019 was 82.76 per 100 000.Over the study period,the incidence rate of AMI increased by 6.05%for men(APC=1.30%,95%CI:0.56%to 2.02%)but decreased by 11.80%for women(APC=-3.10%,95%CI:-4.54%to-1.68%),resulting a steady trend for AMI crude incidence rate for the overall population.The overall ASIR of AMI declined by 16.59%(APC=-4.32%,95%CI:-5.32%to-3.34%)from 113.68 per 100 000 in 2015 to 94.82 per 100 000 in 2019.The ASIR of AMI declined by 11.04%(APC=-2.72%,95%CI:-3.78%to-1.67%)for men,23.96%(APC=-6.56%,95%CI:-8.57%to-4.58%)for women,12.57%(APC=-3.08%,95%CI:-6.01%to-0.08%)for the urban areas,and 19.24%(APC=-5.18%,95%CI:-10.19%to 0.03%)for rural areas respectively.The incidence rate of AMI increases gradually with age in both men and women.The incidence of AMI in urban men of 35-44 and 45-54 year age groups increased by 77.16%(APC=13.52%,95%CI:3.29%to 24.57%)and 26.36%(APC=5.71%,95%CI:-0.95%to 12.68%)over time.However,the incidence of AMI fell in the population above 65 year old,by 26.58%(APC=-6.68%,95%CI:-11.98%to-1.01%),19.85%(APC=-5.64%,95%CI:-11.57%to 0.65%)and 14.53%(APC=-4.44%,95%CI:-7.75%to-1.04%)in the 65-74 year age,75-84 year age and≥85year age groups respectively from 2015 to 2019.The mortality to incidence ratio of AMI was 0.58 over time,higher in women than in men,and higher in rural areas than in urban areas.The M/I ratio of AMI decreased from 0.62 in 2015 to 0.52 in 2019(APC=-4.28%,95%CI:-5.75%to-2.83%).There was a declined trends in M/I of AMI in urban residents of both male and female,and in the rural male residents(all P<0.05),while a steady trend in the rural female residents(P>0.05). Conclusions:The overall incidence of AMI remains steady during 2015 to 2019 in the national surveillance areas in China.Yet,downward trends in elder and female residents and increased trend in middle-aged urban males in AMI incidence are observed.The mortality of AMI in these period are age,sex and urban-rural dependent.Targeted mitigation strategies on AMI prevention and treatment need to be strengthened to reduce its incidence and mortality.

Objective:To assess the trends in case-fatality rates for acute myocardial infarction (AMI) in China from 2015 to 2019.Methods:This study employed a population-based surveillance. Data from the China Registry of Acute Cardiovascular Event (China RACE) were utilized, including AMI cases reported by Grade Ⅱ and Grade Ⅲ hospitals at the disease surveillance sites across China from January 1 st 2015 to December 31 st 2019. The 28-day mortality outcome for reported AMI events was obtained by linking to the national death certificate registry system. The study analyzed the overall and age-standardized case-fatality rates, as well as their annual percent change (APC), during the study period, stratified by gender, age, and region. Results:The overall 28-day case fatality rate for AMI was 28.97% (22 532/77 764) from 2015 to 2019. The age-standardized case-fatality rate for AMI declined significantly from 37.53% in 2015 to 18.58% in 2019, with an APC of -14.33% ( P=0.018). We observed a significant downward trend in case-fatality rates of AMI in both genders (both P<0.05). Among males, the case-fatality rate decreased more steeply in younger males compared to elder counterparts. The most marked decreases were seen in males aged<35 years and 35 to 44 years, with APC of -27.63% ( P=0.007) and -22.65% ( P=0.004), respectively. In females, we observed a relatively stable decrease in case-fatality across age groups. The age-standardized case-fatality rate of AMI in eastern and central China decreased significantly from 2015 to 2019, with the APC of -19.22% ( P=0.006) and -15.62% ( P=0.032) respectively. However, the age-standardized case-fatality rate of AMI in western China remained stable ( P=0.227). Conclusions:The prognosis of AMI has considerably improved from 2015 to 2019 in China, regardless of ages and gender. Inequality in case-fatality rates among geographic regions highlights the need for targeted strategies in AMI prevention in western regions.

Objective:To evaluate changes in operational effectiveness after the implementation of ambulatory surgical management in pars plana vitrectomy (PPV).Methods:A retrospective clinical study. 17 528 surgeries in 10 895 eyes of 10 895 patients who underwent minimally invasive PPV on an ambulatory and/or inpatient basis at Tianjin Medical University Eye Hospital from August 2015 to June 2023 were included in this study. Among them, 5 346 eyes in 5 346 cases were male; 5 549 eyes in 5 549 cases were female. The age ranged from 0 to 95 years, with the mean age of (57.74±13.15) years. 6 381 surgeries in 3 615 eyes from August 2015 to December 2018 (the initial period of day surgery) were used as the control group; 11 147 surgeries in 7 280 eyes from January 2019 to June 2023 (the expanded period of day surgery) were used as the observation group. According to the management mode of ambulatory surgery, the observation group was subdivided into the decentralized management group (January 2019 to December 2020) and the centralized management group (January 2021 to June 2023), with 2 905 and 4 375 eyes and 4 646 and 6 501 surgeries, respectively. Changes in the percentage of day surgery, average hospitalization days, and average unplanned reoperation rate were compared. The Mann-Whitney U test was used to compare numerical variables between groups; the chi-square test or Fisher's exact test was used to compare categorical variables. Results:The number of cases of daytime PPV performed in the observation group and control group was 7 852 (70.44%, 7 852/11 147) and 24 (0.38%, 24/6 381) cases, respectively, and the average hospitalization days were 1 (1) and 5 (3) d. Compared with the control group, the observation group had a significantly higher percentage of day surgery ( χ2=8 051.01) and a considerably lower mean hospitalization day ( Z=4 536 844.50), and the differences were statistically significant ( P<0.000 1). The mean hospitalization days in the decentralized and centralized management groups were 2 (3) and 1 (0) d, respectively, and unplanned reoperations were 34 (0.73%, 34/4 646) and 171 (2.63%, 171/6 501) eyes, respectively. Compared with the decentralized management group, average hospitalization days was significantly lower ( Z=1 436.94) and unplanned reoperation rate was significantly higher ( χ2=54.10) were significantly lower in the centralized management group, both of which were statistically significant ( P<0.000 1). Conclusion:PPV ambulatory management model can significantly reduce the average hospitalization day, but also results in higher rates of unplanned reoperations.