Stroke is one of the leading causes of disability and mortality worldwide. Research into its mechanisms and the development of therapeutic strategies heavily rely on animal models that accurately replicate the pathological features of human disease. An ideal animal model for stroke should not only reproduce the neurological deficits and pathological changes observed in clinical patients but also demonstrate good reproducibility and translational value. This review focuses on the preparation and evaluation methods of ischemic stroke animal models. Firstly, it elaborates on the selection criteria, advantages, and disadvantages of experimental animals, including rodents (rats, mice) and non-rodents (non-human primates, miniature pigs, rabbits, zebrafish). Secondly, it provides a detailed overview of the modeling principles, key procedures, and application scopes for ischemic stroke models and hemorrhagic stroke models. Furthermore, the review summarizes advances in the applications of emerging technologies—including gene editing [e.g., clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing], multimodal imaging (e.g., two-photon microscopy, photoacoustic imaging), artificial intelligence, optogenetics, 3D bioprinting, organoid models, and multi-omics–in model optimization, precise assessment, and mechanistic investigation. Finally, based on a systematic analysis of relevant domestic and international literature from 2019 to 2024, this review discusses model selection strategies based on research objectives, a multidimensional evaluation system encompassing behavioral, imaging, and molecular pathological assessments, and envisions future directions involving technological integration to achieve model precision and individualization. This article aims to provide a comprehensive methodological reference to help researchers select appropriate animal models of stroke according to specific scientific questions.

Objective:To investigate the effect of early protein supplementation on the clinical outcomes of elderly ICU patients with critically ill.Methods:The study was a post-hoc analysis of a multicenter, cluster randomized controlled trial (NEED trial), which aimed to evaluate the impact of feeding protocol on nutritional implementation and outcomes in ICU patients. It was planned to include elderly patients aged ≥70 years from the NEED trial, and patients who had not started nutritional therapy by the Day 3 after enrolment, stayed in the ICU less than 7 days, missing the primary outcome were excluded. The primary outcome of this study was 28-day mortality of enrolment. Patients were categorized into Q1 (<0.6 g/kg/d), Q2 (0.6-0.83 g/kg/d), and Q3 (≥0.83 g/kg/d) groups according to the tertiles of protein supply. The log-rank test was used to compare the Kaplan-Meier survival curves for 28-day mortality. The associations between different protein groups and 28-day mortality were tested by Cox proportional hazards regression models. Subgroup analysis was conducted in patients with high (mNUTRIC score≥5) nutritional risk or patients with baseline acute kidney injury.Results:A total of 789 elderly (≥70 years) patients was included in the study, with a mean protein amount of 0.69 (0.53, 0.91) g/(kg·d) during days 3-7 after ICU admission, and mean protein amounts in the Q1 low-protein group, the Q2 medium-protein group, and the Q3 high-protein group were 0.46 (0.36, 0.53), 0.69 (0.63, 0.76), and 1.03 (0.91, 1.23) g/(kg·d), respectively. The results showed that the medium protein group associated with lower 28-day mortality compared to the high protein group, and the association between the medium protein group and lower 28-day mortality still held after controlling for possible confounders by Cox multivariate regression analysis. In the high-nutritional risk subgroup (mNUTRIC≥5), a significant association was also found between the medium protein group and lower 28-day mortality.Conclusions:Early high protein supply are not beneficial for elderly ICU patients by this large sample size post-hoc analysis, and medium protein supply associate with lower 28-day mortality compared with the high protein group. This study may provide a theoretical basis for the optimal dose of early protein supply in elderly ICU patients, as well as a reference for clinical implementation.

ObjectiveEstablish an animal model of preeclampsia in goats, collect data on various physiological indicators and maternal biological characteristics of the disease model to compare with clinical feature of the disease in humans, provide reference for the prevention and treatment of preeclampsia in humans.Methods Twenty-three goats bred in Chongming district were divided into three groups: Control group, no surgical procedure was performed on animals of this group; sham group, ewes in this group underwent the sham operation on the 100±5th day of gestation, and only the uterine artery was exposed and dissociated; surgical group, a silver vascular clamp was clipped on one side of the uterine body artery of the ewe to narrow the inner diameter of the artery at the same gestation period (100±5) days. Heart rate and hindlimb blood pressure were continuously monitored in control and surgical ewes from 100 to 140 d of gestation, and blood flow data within the lateral branches of the uterine arteries of ewes in the sham group were collected using a hemodynamometer in combination with a hemodynamic probe and an animal physiological signal collector, as well as changes in blood flow within the uterine arteries in the lateral branches of the uterine arteries of the surgical group before and after placement of vascular clips in the surgical ewes. At the expected date of delivery, jugular vein blood was taken from ewes for routine blood test, creatinine, urea nitrogen, and blood ion analysis; urine was also collected from ewes in each group for analysis of urinary protein and urinary creatinine. All experimental groups were subjected to cesarean section on the 140±5th day of gestation in ewes, and then liver, kidney, uterus and placenta tissues were taken from ewes in each group and stained with HE for pathological observation. Results After 15 minutes of preeclampsia modeling surgery, blood flow volume remained stable in the vessel stenosis segment and the volume differential was relatively reduced in comparison to the control group and sham group (P＜0.05，P＜0.01). Compared with the control group, the ewes in the surgical group showed prenatal changes such as increased serum osmolality, decreased hemoglobin, increased blood glucose and urea nitrogen values, as well as increased levels of calcium, sodium, and chloride ions (all P<0.05) and proteinuria, with urinary creatinine and urinary protein-creatinine ratios were significantly higher than those in the control group and sham group (all P<0.05). The elastic lamina of the uterine body arteries on the operated side of the animals in the surgical group was thicker than that on the opposite side, but the structure was loose. The placenta on the operated side showed pathological changes such as cell interstitial swelling and inflammatory cell infiltration. The above physiological index characteristics were more consistent with the clinical features of human preeclampsia disease.Conclusion In this experiment, we successfully constructed a goat preeclampsia model and obtained data on relevant physiological indexes of this model, which further verified the correlation between preeclampsia disease and uterine artery lesions.

Objective:To analyze the molecular epidemiology and virulence characteristics of polymyxin-resistant Klebsiella pneumoniae ( K. pneumoniae). Methods:From 2011 to 2016, 1 376 strains of K. pneumoniae were isolated from various clinical specimens of hospitalized patients in First Affiliated Hospital of Wenzhou Medical University. Agar dilution method was used to screen out the polymyxin-resistant strains.Polymerase chain reaction (PCR) was used to detect the genes related to polymyxin resistance, and real-time fluorescence quantitative PCR was used to detect the relative mRNA expression level of drug resistant genes. Pulsed-field gel electrophoresis, multilocus sequence typing and Galleria mellonella larvae infection model were performed to analyze the molecular epidemiological and virulent characteristics. Results:A total of 14 strains (1.02%) of polymyxin-resistant K. pneumoniae were detected among 1 376 K. pneumoniae isolates. Subsequent sequencing identified mutations leading to amino-acid changes (K2E, F28C) in MgrB of 10 isolates and D150G in PhoQ of nine isolates, and genes such as mcr and crrB were not detected in all drug-resistant strains. Compared with standard strains, the relative expression levels of pmrH and pmrD mRNA of these drug resistant strains were increased. Analysis of the molecular epidemiology indicated that the 14 drug-resistant strains were divided into nine clones. Galleria mellonella larvae infection model revealed polymyxin-resistant K. pneumoniae isolates had higher virulence. Conclusions:Polymyxin-resistant K. pneumoniae has mutations in mgrB and phoQ genes, and mgrB mutation may play a key role in the change of virulence profiles. The homology among the polymyxin-resistant K. pneumoniae stains in this study is low.

An in vitro blood-brain barrier (BBB) model is critical for enabling rapid screening of the BBB permeability of the drugs targeting on the central nervous system. Though many models have been developed, their reproducibility and renewability remain a challenge. Furthermore, drug transport data from many of the models do not correlate well with the data for in vivo BBB drug transport. Induced-pluripotent stem cell (iPSC) technology provides reproducible cell resources for in vitro BBB modeling. Here, we generated a human in vitro BBB model by differentiating the human iPSC (hiPSC) line GM25256 into brain endothelial-type cells. The model displayed BBB characteristics including tight junction proteins (ZO-1, claudin-5, and occludin) and endothelial markers (von Willebrand factor and Ulex), as well as high trans-endothelial electrical resistance (TEER) (1560 Ω.cm ± 230 Ω.cm) and γ-GTPase activity. Co-culture with primary rat astrocytes significantly increased the TEER of the model (2970 Ω.cm to 4185 Ω.cm). RNAseq analysis confirmed the expression of key BBB-related genes in the hiPSC-derived endothelial cells in comparison with primary human brain microvascular endothelial cells, including P-glycoprotein (Pgp) and breast cancer resistant protein (BCRP). Drug transport assays for nine CNS compounds showed that the permeability of non-Pgp/BCRP and Pgp/BCRP substrates across the model was strongly correlated with rodent in situ brain perfusion data for these compounds (R = 0.982 and R = 0.9973, respectively), demonstrating the functionality of the drug transporters in the model. Thus, this model may be used to rapidly screen CNS compounds, to predict the in vivo BBB permeability of these compounds and to study the biology of the BBB.

Objective To investigate the appropriate age of primary and secondary school students for cardiopulmonary resuscitation (CPR) training.Methods A total of 437 students aged 9-15 years at 3 to 6 grade in the primary schools or 1 to 2 grade in the secondary schools were selected from 2 Yuyao primary and secondary schools by stratified random sampling between March 2017 and January 2018.The numbers of students with the age of 9,10,11,12,13,14 and 15 y were 61,62,66,64,63,63 and 58,respectively.All students received chest compression training provided by Yuyao emergency department People's Hospital according to the 2015 Cardiopulmonary Resuscitation Guidelines.The training included 30 min theoretic teaching and 6 min practice in the simulator.The quality of chest compression performed by students was assessed;the depth,rate,position and retention of chest compression were recorded.Results The mean depth of chest compression in the students aged 9-15 years was 3.8,4.1,4.6,5.1,5.2,5.6 and 5.6 cm,respectively;the accuracy rate was 24.6％(14/61),25.8％ (16/62),50.2％ (33/66),70.5％ (45/64),79.4％(50/63),88.9％(56/63) and 91.4(53/58),respectively.Compared with the students aged 9-11 years,the mean depth of chest compression was significantly increased and accuracy rate was significantly improved in the students aged 12-15 years (Compared with 9-y students,t=-8.936,-9.502,-10.640 and-11.370;x2=35.019,47.599,63.013 and 65.671;compared with 10-y students,t=-6.927,-8.179,-10.70 and-11.047;x2=24.977,35.967,50.916 and 52.727;compared with 1 1-y students,t=-3.095,-4.177,-6.785 and-6.995;x2=5.586,12.114,22.786 and 24.870;all P＜0.05).The mean rate of chest compression was 110-116/min and its accuracy rate was 86.4％-95.2％;the accuracy rate of chest compression position was 90.9％-96.8％ in all students,there were no significant differences among the 7 groups.The mean retention rate of chest compression in the 7 groups was 81.3％(122/150),67.3％(101/150),64.7％ (94/150),48.0％(72/150),48.7％(73/150),33.3％(50/150) and 27.3％(41/150),respectively.Compared with the students aged 9-11 years,the mean retention rate of chest compression was significantly decreased in the students aged 12-15 years (compared with the 9-y students,x2=36.472,35.179,70.64 and 119.92;compared with 10-y students,x2=11.483,10.728,34.682 and 72.150;compared with 11-y students,x2=6.528,5.927,25.855 and 59.11;all P＜0.05).Correlation analysis showed that the depth (r=0.96,0.89,0.91 and 0.86;P＜0.01) and retention rate (r=-0.99,-0.90,-0.93 and-0.86;all P＜0.01) of chest compression were significantly associated with the age,body weight,height and body mass index of students.Conclusion The students with an age of 12 years or more are able to effectively perform chest compression;thus,12 years and above might be the appropriate age for CPR training.

Objective: In order to understand the current situation and influencing factors of teaching quality in a Medical University in Yunnan, thus improving the teaching quality of the teachers. Methods: The self-made evaluation forms for teachers' teaching quality which include 9 first-level indicators were adopted. In December 2016, a survey was conducted on some students from grade 2013 to 2016 about the teachers who gave them lectures from September to December 2016, involving 7 different majors, 23 teachers and 18 courses. SPSS 21.0 was used for data analysis. Enumeration data were described by frequencies and percentages. Univariate and multivariate Logistic regression analyses were used for satisfaction. Results: There were 1 542 valid questionnaires collected in this survey. Univariate Logistic regression analysis showed that there were 6 factors related to the overall satisfaction of teachers' classroom teaching quality, which were "whether the courses are professional (OR=1.430)", "curriculum difficulty (OR=1.731)", "curriculum progression (OR=1.620)", "learning time after class (OR=1.141)", "interest before class (OR=1.565)", and "interest after class (OR=2.291)", respectively. Conclusion "courses are professional", "courses are too easy", and "the progression of courses is slow " are favorable factors for high satisfaction degree, while "courses are set up by the departments that the students belong", "learning time used after class is too short", and "interest for study is decreased after class" may be unfavorable factors for high satisfaction degree.

Objective: To evaluate the in vitro antibiotic effects of colistin combined with meropenem, levofloxacin or fosfomycin against colistin-heteroresistant Acinetobacter baumannii (A.baumannii). Methods: A total of 576 A. baumannii clinical isolates were collected from the First Affiliated Hospital of Wenzhou Medical University from 2014 to 2015. Minimal inhibitory concentrations (MICs) of colistin against A. baumannii were detected by broth dilution method. Colistin-heteroresistant A. baumannii isolates were screened using population analysis profiles (PAPs). MICs of colistin combined with meropenem, levofloxacin or fosfomycin, and the four drugs used alone against colistin-heteroresistant A. baumannii were detected by checkerboard method and broth dilution method. Fractional inhibitory concentration index (FICI) was calculated to evaluate antibiotic effects. Results: None of the 576 A. baumannii isolates was resistant to colistin as indicated by the broth dilution method. Nine colistin-heteroresistant A. baumannii isolates were identified using PAPs. Compared with the MICs of colistin used alone, the MICs of colistin used in combination with meropenem, levofloxacin or fosfomycin against colistin-heteroresistant isolates were all decreased. Colistin-meropenem combination showed synergistic (55.6%), additive (33.3%) and indifferent effects (11.1%), but no antagonistic effect. Colistin-levofloxacin combination showed synergistic (55.6%), additive (22.2%) and indifferent effects (22.2%), but no antagonistic effect. Colistin-fosfomycin combination showed synergistic (77.8%) and additive (22.2%) effects, but no indifferent or antagonistic effect. Conclusion In vitro use of colistin in combination with meropenem, levofloxacin or fosfomycin has synergistic and additive antibacterial effects against colistin-heteroresistant A. baumannii. Combinations of colistin-meropenem and colistin-levofloxacin have fewer indifferent effects and no antagonistic effect.

Objective To investigate the clinical feasibility of using gemstone spectral image (GSI)technique to reduce the individual dose of contrast agent in performing lower extremity angiography.Methods Among the consecutive 75 patients receiving lower extremity CTA examination,60 patients were enrolled in this study.According to the scanning method and contrast dosage,the 60 patients were prospectively and randomly divided into the conventional CT routine-dose group (n=20),GSI routine-dose group (n=20) and GSI low-dose group (n=20).In the conventional CT routine-dose group and GSI routine-dose group,the dosage of contrast medium was 2 ml/kg and the injection rate was 3.5 ml/s.Scanning with 120 kVp and reconstruction with filter back projection (FBP) technology were used in the conventional CT routine-dose group,while scanning with spectrum mode and reconstruction with 50％ adaptive statistical iterative reconstruction (ASiR) technology were used in the GSI routine-dose group.In the GSI low-dose group,the dosage of contrast medium and the injection rate were 1.6 ml/kg and 2.8 ml/s respectively,and scanning with spectrum mode and reconstruction with 50％ ASiR technology were adopted.The target vessel CT values of three sets of images were determined,the contrast-to-noise ratio (CNR),the background noise (BN),the dose length product (DLP),the effective dose (ED),the iodine intake dose and the injection rate were calculated and compared between each other among the three groups.Single factor variance analysis and Bonferroni correction method were used to statistically analyze the results.By using Kruskal-Wallis test,the subjective scores of three sets of images were compared between each other among the three groups.Results According to exclusion criteria,60 patients were finally enrolled in this study.The images of GSI routine-dose group had the best CT value,CNR and BN (P＜0.01),but there was no statistically significant difference in BN between GSI routine-dose group and GSI low-dose group (P＞0.05).Both the CT value and CNR,except anterior tibial artery (P=0.162 and P=0.376 respectivcly),in the GSI low-dose group were higher than those in the conventional CT routine-dose group (P＜0.05).The ED values of both GSI groups were lower than that of the conventional CT routine-dose group (P＜0.01),although no statistically significant difference in ED value existed between the two GSI groups (P＞0.05).The iodine intake dose and injection rate of the GSI lowdose group were strikingly lower than those of the conventional CT routine-dose group and the GSI low-dose group (both P＜0.01).The subjective scores of three sets of images evaluated by two physicians were 82,95,90 points and 80,96,89 points respectively,the differences were statistically significant (H=14.954,P＜0.01;H=17.726,P＜0.01).GSI routine-dose group had the best image quality,and the image quality of GSI low-dose group was superior to that of conventional CT routine-dose group (P＜0.05).Conclusion Compared with conventional spiral CT scanning,gemstone optimal monochromatic imaging technique can obtain more optimal CNR and image quality,meanwhile,the radiation dose can be remarkably reduced while the imaging quality of lower extremity angiography can meet the requirements of making clinical diagnosis,moreover,individual intake of iodine can be reduced.

Objective To evaluate the feasibility and safety of prostatic artery embolization (PAE)via transradial approach in treating prostatic hyperplasia.Methods The clinical data of 18 patients with prostatic hyperplasia,who received C-arm CT-guided PAE via left or right radial artery access,were retrospectively analyzed.The following indexes were recorded:arterial spasm and injury of upper limbs,incidence of puncture point bleeding,postoperative radial artery pulse and congestion,blood supply and nerve injury of fingers,the surgical success rate,incidence of perioperative cerebral vascular complications,operation time,radiation dose and clinical curative effect.Results Among the 18 patients,PAE via left radial artery access was employed in 14,and PAE via right radial artery access was performed in 4.Bilateral PAE was carried out in 16 patients,and only unilateral PAE was able to be successfully accomplished in 2 patients as the prostatic artery opening of the other side was tortuous with stenosis.After PAE,decreased radial pulse was observed in one patient and ultrasound examination revealed decreased blood flow.The operation time ranged from 96 min to 245 min.The radiation dose received by the patient varied from 2435 mGy to 4958 mGy with a mean of (3342±156) mGy,which was not significantly different from the radiation dose received by the patients who underwent PAE via femoral artery access during the same study period (P=0.1167).Conclusion In treating prostatic hyperplasia,PAE by using transradial approach is clinically safe and technically feasible.