Randomized controlled trial (RCT) are considered the "gold standard" for evaluating the causal effects of interventions on outcome measures. However, due to high research costs and ethical constraints, conducting RCT in clinical practice, especially in the surgical field, faces numerous challenges such as difficulties in subject recruitment, implementation of blinding, and standardization of interventions. In such cases, using real-world data to perform causal inference under the framework of target trial emulation (TTE), based on the principles of RCT design, helps to identify and reduce biases arising from design flaws in traditional observational studies, such as immortal time bias, confounding, selection bias, or collider bias. This approach can produce high-quality evidence comparable to that of RCT, thereby enhancing the clinical guidance value of real-world data studies. However, TTE has limitations, such as the inability to completely eliminate confounding, high quality requirements for source data, and the current lack of reporting standards. Therefore, researchers should be fully aware of these limitations to avoid making incorrect causal inferences. This article intends to provide an overview of the TTE framework, implementation points, application scope, application cases, and advantages and disadvantages of the framework.

Sepsis is a life-threatening organ dysfunction caused by host's dysfunctional response to infection, which is an important cause of health threat worldwide. Recent studies have found that the metabolic disorder of iron homeostasis is closely related to sepsis. The body has a highly accurate iron metabolism and regulation system to prevent iron deficiency or iron overload. The imbalance of intracellular iron homeostasis is now considered to be one of the important mechanisms of sepsis and may be a new target for clinical intervention.

Objective·To develope an auditory brainstem implant(ABI)vocoder based on cochlear implant(CI)vocoder characteristics and ABI electrode array topology,and to verify its reliability.Methods·An"n-of-m"coding strategy CI/ABI vocoder was constructed based on MATLAB.Within each frame,only the envelopes of the n channels with the highest energy were selected.The interaction coefficient(IC)(range:1?3),channel numbers(range:5?22),and electrode array topology(CI/ABI)were adjustable parameters,allowing for the synthesis of simulated speech.Psychoacoustic evaluation was employed,recruiting normal hearing subjects to perform closed-set simulated phoneme perception.The phoneme recognition accuracy(20 vowel questions/condition,11 consonant questions/condition)was compared with the corresponding conditions of CI and ABI from reference literature to determine the IC value of the vocoder and verify its reliability.Results·The vocoder successfully synthesized all test stimuli.In the closed-set CI-simulated speech recognition,the simulated vowel and consonant recognition accuracy for IC2 and IC3 conditions showed no significant difference compared to the accuracy reported in the CI reference literature(P＞0.05).The difference in vowel and consonant accuracy between IC2 and the literature was smaller than that between IC3 and the literature(vowel|d|=1.6%vs.20%,consonant|d|=8.4%vs.9.9%),thus determining the optimal interaction coefficient of this model as 2.Subsequently,when modifying the electrode array topology to ABI,it was found that the simulated phoneme recognition accuracy for a 16-channel ABI was significantly lower than that for the 16-channel CI group,consistent with the reported literature.The simulated vowel and consonant accuracy within the 5?8 channel range for ABI showed no significant difference(P＞0.05),also aligning with the trend reported in the literature.Conclusion·A CI/ABI vocoder based on"n-of-m"coding strategy is established and the optimal IC is determined.The established ABI encoder has been evaluated for high reliability through psychoacoustic experiments.It provides suitable technical means for validating ABI-specific coding strategies.

Objective This study aimed to develop an experimental animal platform for evaluating the feasibility and safety of intelligent acupuncture robots and to lay the foundation for further research.Methods Six 2-month-old Guangxi Bama miniature pigs were used as experimental subjects for acupuncture verification after anesthesia.First,manual acupuncture verification was carried out.Six acupoints were selected for each experimental animal and the needles were left for 20 min after the lifting,inserting,and twisting manipulation.Before and after controls were included.The experiment was carried out for 28 days,and each experiment was conducted once every 2 days for a total of 10 times.After verification of manual acupuncture,a point 10 mm from each of the six selected acupoints was selected,with a total of 12 points,and acupuncture operations were carried out on the experimental animals using the intelligent acupuncture module of the acupuncture robot at different frequencies and angles,to further verify the stability and feasibility of the animal platform.Results Routine safety-related blood indicators and blood biochemistry indicators after the procedure were normal and stable compared with those before the procedure.The average heart rate of the animals was 124 beats/min,the average blood pressure was 87/36 mmHg,and the average body temperature of was 36℃at a room temperature of 25℃,with no significant change in body temperature during and after the experiment.On the basis of this experimental platform,acupuncture manipulation using the intelligent acupuncture module of the acupuncture robot was completed successfully,with no abnormalities related to acupuncture such as bending,breaking,or stagnation of needles during the experimental process,and the experimental animals showed no obvious abnormalities.Conclusions This study established a stable experimental animal platform for evaluating the feasibility and safety of acupuncture carried out by intelligent acupuncture robots,based on the existing experimental method of miniature pigs.These result lay a foundation for further research related to the use of intelligent acupuncture robots.

Spinocerebellar ataxias are a high clinically and genetically heterogeneous group of neurodegenerative disorders, usually belongs to autosomal dominant hereditary cerebellar ataxia. Spinocerebellar ataxia type 5 (SCA5) is one of the extremely rare subtypes and caused by heterozygous mutation of SPTBN2 gene. A case of infant-onset SCA5 patient is reported, mainly manifested as global developmental delay, ataxia and dysarthria, carrying the heterozygous missense variant c.1438C>T (p. Arg480Trp) in the SPTBN2 gene. This mutation may have an important impact on functional regions of the β-Ⅲ spectrin, leading to the occurrence of disease.

Objective: To evaluate the effects of ferroptosis in hippocampal neurons on cognitive dysfunction in rats with sepsis-associated encephalopathy (SAE) and its potential molecular mechanisms. Methods: ① Screening experiment of SAE modeling conditions: 42 healthy male Sprague-Dawley (SD) rats were divided into normal saline (NS) control group (n = 6) and lipopolysaccharide (LPS) 5, 15, 30 mg/kg groups (each n = 12) according to the random number table method. The SAE modeling conditions were determined by survival and the changes in mean arterial pressure (MAP) and heart rate (HR) within 72 hours, the percentage of stiffness status, the levels of serum inflammatory factors including interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), neuron specific enolase (NSE, a marker of neuronal injury), serum iron and lactic acid (Lac) contents, and the morphological changes in CA1 of hippocampus after 72 hours. ② Deferoxamine (Def) intervention experiment: according to the results of screening experiments, 28 healthy male SD rats were divided into NS control group (n = 8), SAE group (n = 10) and Def+SAE group (n = 10) according to the random number table method. In the Def+SAE group, 100 mg/kg Def was injected intraperitoneally 12 hours before the modeling, once every 12 hours, with a total of 7 times; the rats in the NS control group and SAE group were injected with the same amount of NS. Then the cognitive function of rats was evaluated by fear conditioning test for the percentage of stiffness status; serum IL-6, TNF-α and NSE levels were determined by enzyme-linked immunosorbent assay (ELISA); the levels of serum Lac, serum iron and hippocampal malondialdehyde (MDA) and iron contents were determined by chemical colorimetric; the protein expressions of nuclear factor E2 related factor 2 (Nrf 2), glutathione peroxidase 4 (GPX4) and NAPDH oxidase 1 (NOX1) in hippocampus were determined by Western Blot; morphological changes in hippocampal CA1 were observed after hematoxylin and eosin (HE) staining. Results: ① Compared with the NS control group, intraperitoneal injection of 15 mg/kg LPS could significantly reduce the MAP and HR as time prolonged, and the reduction was most significant at 72 hours. The 72-hour survival rate was significantly reduced and cognitive function was impaired. The levels of serum IL-6, TNF-α, Lac and NSE were increased while the serum iron content was decreased significantly. The morphology of vertebral cells in hippocampal CA1 was irregular and some of the cells were obviously vacuolated. In the LPS 5 mg/kg group, there were no significant changes in vital signs, inflammation, organ function or cognitive dysfunction, while the symptoms of septic shock were apparent in the LPS 30 mg/kg group. Therefore, SAE model was reproduced by intraperitoneal injection of 15 mg/kg LPS for 72 hours. ② Compared with the NS control group, the percentage of stiffness in the SAE group was significantly reduced. The levels of serum IL-6, NSE and hippocampal MDA, iron were significantly increased. The serum iron contents and hippocampal Nrf 2 and GPX4 protein expressions were significantly reduced, while the hippocampal NOX1 protein expression was significantly increased. The morphology of vertebral cells in hippocampal CA1 was irregular and the cytoplasm was deeply stained. The results indicated that the level of oxidative stress in the hippocampus of SAE rats was increased, the neuron degenerations were obvious, and the cognitive function of rats were impaired. Compared with the SAE group, the percentage of stiffness in the Def+SAE group was significantly increased [(63.4±6.4)% vs. (47.6±6.0)%, P < 0.05]. The levels of serum IL-6, NSE, iron and hippocampal MDA, iron were significantly reduced [serum IL-6 (ng/L): 73.14±8.31 vs. 99.86±12.37, serum NSE (μg/L): 3.67±0.51 vs. 5.92±0.79, serum iron (mg/L): 68.43±8.12 vs. 134.60±15.63, hippocampal MDA (mol/g): 4.62±0.90 vs. 6.62±0.84, hippocampal iron (μg/g): 155.32±17.86 vs. 221.54±27.54, all P < 0.05]. The hippocampal protein expressions of Nrf 2 and GPX4 were significantly increased [Nrf 2/β-actin: 0.41±0.07 vs. 0.18±0.03, GPX4/β-actin: 0.74±0.09 vs. 0.40±0.06, all P < 0.05] while the hippocampal NOX1 protein expression was significantly reduced (NOX1/β-actin: 0.62±0.08 vs. 1.11±0.16, P < 0.05). The vertebral cells was significantly improved as compared with the SAE group. These findings showed that the oxidative stress level in hippocampus of the Def+SAE group was reduced, neuron degeneration was significantly alleviated, and the cognitive function of the rats was significantly improved. Conclusions The cognitive function of rats with SAE was significantly impaired, the hippocampal neurons were obviously damaged and ferroptosis was increased. Def pretreatment could significantly reduce iron deposition and ferroptosis in hippocampal neurons of SAE rats and improve cognitive dysfunction, which may be related to activation of Nrf 2/GPX4 signaling pathway.

OBJECTIVE: To evaluate the effects of ferroptosis in hippocampal neurons on cognitive dysfunction in rats with sepsis-associated encephalopathy (SAE) and its potential molecular mechanisms. METHODS: (1) Screening experiment of SAE modeling conditions: 42 healthy male Sprague-Dawley (SD) rats were divided into normal saline (NS) control group (n = 6) and lipopolysaccharide (LPS) 5, 15, 30 mg/kg groups (each n = 12) according to the random number table method. The SAE modeling conditions were determined by survival and the changes in mean arterial pressure (MAP) and heart rate (HR) within 72 hours, the percentage of stiffness status, the levels of serum inflammatory factors including interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), neuron specific enolase (NSE, a marker of neuronal injury), serum iron and lactic acid (Lac) contents, and the morphological changes in CA1 of hippocampus after 72 hours. (2) Deferoxamine (Def) intervention experiment: according to the results of screening experiments, 28 healthy male SD rats were divided into NS control group (n = 8), SAE group (n = 10) and Def+SAE group (n = 10) according to the random number table method. In the Def+SAE group, 100 mg/kg Def was injected intraperitoneally 12 hours before the modeling, once every 12 hours, with a total of 7 times; the rats in the NS control group and SAE group were injected with the same amount of NS. Then the cognitive function of rats was evaluated by fear conditioning test for the percentage of stiffness status; serum IL-6, TNF-α and NSE levels were determined by enzyme-linked immunosorbent assay (ELISA); the levels of serum Lac, serum iron and hippocampal malondialdehyde (MDA) and iron contents were determined by chemical colorimetric; the protein expressions of nuclear factor E2 related factor 2 (Nrf2), glutathione peroxidase 4 (GPX4) and NAPDH oxidase 1 (NOX1) in hippocampus were determined by Western Blot; morphological changes in hippocampal CA1 were observed after hematoxylin and eosin (HE) staining. RESULTS: (1) Compared with the NS control group, intraperitoneal injection of 15 mg/kg LPS could significantly reduce the MAP and HR as time prolonged, and the reduction was most significant at 72 hours. The 72-hour survival rate was significantly reduced and cognitive function was impaired. The levels of serum IL-6, TNF-α, Lac and NSE were increased while the serum iron content was decreased significantly. The morphology of vertebral cells in hippocampal CA1 was irregular and some of the cells were obviously vacuolated. In the LPS 5 mg/kg group, there were no significant changes in vital signs, inflammation, organ function or cognitive dysfunction, while the symptoms of septic shock were apparent in the LPS 30 mg/kg group. Therefore, SAE model was reproduced by intraperitoneal injection of 15 mg/kg LPS for 72 hours. (2) Compared with the NS control group, the percentage of stiffness in the SAE group was significantly reduced. The levels of serum IL-6, NSE and hippocampal MDA, iron were significantly increased. The serum iron contents and hippocampal Nrf2 and GPX4 protein expressions were significantly reduced, while the hippocampal NOX1 protein expression was significantly increased. The morphology of vertebral cells in hippocampal CA1 was irregular and the cytoplasm was deeply stained. The results indicated that the level of oxidative stress in the hippocampus of SAE rats was increased, the neuron degenerations were obvious, and the cognitive function of rats were impaired. Compared with the SAE group, the percentage of stiffness in the Def+SAE group was significantly increased [(63.4±6.4)% vs. (47.6±6.0)%, P < 0.05]. The levels of serum IL-6, NSE, iron and hippocampal MDA, iron were significantly reduced [serum IL-6 (ng/L): 73.14±8.31 vs. 99.86±12.37, serum NSE (μg/L): 3.67±0.51 vs. 5.92±0.79, serum iron (mg/L): 68.43±8.12 vs. 134.60±15.63, hippocampal MDA (mol/g): 4.62±0.90 vs. 6.62±0.84, hippocampal iron (μg/g): 155.32±17.86 vs. 221.54±27.54, all P < 0.05]. The hippocampal protein expressions of Nrf2 and GPX4 were significantly increased [Nrf2/β-actin: 0.41±0.07 vs. 0.18±0.03, GPX4/β-actin: 0.74±0.09 vs. 0.40±0.06, all P < 0.05] while the hippocampal NOX1 protein expression was significantly reduced (NOX1/β-actin: 0.62±0.08 vs. 1.11±0.16, P < 0.05). The vertebral cells was significantly improved as compared with the SAE group. These findings showed that the oxidative stress level in hippocampus of the Def+SAE group was reduced, neuron degeneration was significantly alleviated, and the cognitive function of the rats was significantly improved. CONCLUSIONS The cognitive function of rats with SAE was significantly impaired, the hippocampal neurons were obviously damaged and ferroptosis was increased. Def pretreatment could significantly reduce iron deposition and ferroptosis in hippocampal neurons of SAE rats and improve cognitive dysfunction, which may be related to activation of Nrf2/GPX4 signaling pathway.
Animals ; Cognitive Dysfunction ; Ferroptosis ; Hippocampus ; Male ; NF-E2-Related Factor 2/metabolism* ; Neurons ; Rats ; Rats, Sprague-Dawley ; Sepsis ; Sepsis-Associated Encephalopathy ; Signal Transduction ; Tumor Necrosis Factor-alpha

Animal infectious diseases pose a serious and continuing threat to the animal health and cause huge economic losses throughout the world. Vaccination is one of the most effective solutions to prevent and control animal infectious diseases. With the development of biotechnologies and the need for disease prevention and control, the focus of vaccine research has been shifted to the development of safe, efficient, broad-spectrum, low-dose and marker vaccines. Novel vaccines capable of inducing high levels of both humoral and cellular immune responses are promising to provide more efficient protection against animal infectious diseases. This minireview summarizes the development, applications, advantages and disadvantages of new-concept animal vaccines emerging in recent years, including mucosal vaccines, long-acting and fast-acting vaccines, chimeric vaccines, nanoparticle vaccines, and so on. Furthermore, we discuss future directions of the vaccines, in order to provide new insights for animal vaccine development.
Animals ; Communicable Diseases ; Immunity, Cellular ; Nanoparticles ; Vaccination ; Vaccines

OBJECTIVEObesity has been shown to be an indicator of poor prognosis for patients with primary breast cancer. The aim of this study was to clarify the effect of obesity on Chinese women with breast cancer.

METHODSThis is a retrospective analysis of 1699 breast cancer patients. We evaluated the effect of body mass index (BMI) on disease-free survival (DFS) and overall survival (OS) in these patients. BMI was obtained before surgery. Obesity was defined as a BMI ≥ 24. Kaplan-Meier analysis and Log rank test were employed to perform survival analysis. The impact of different characteristics on survival was assessed by using Cox proportional-hazards regression model.

RESULTSIn total 635 (37.4%) patients were obese, while 1 064 (62.6%) were non-obese. Comparing the tumor characteristics in the two groups, the BMI ≥ 24 group showed a higher rate of older age (P < 0.001), postmenopausal status (P < 0.001), increased risk of lymph node metastasis (P = 0.001) and less chances of accepting breast conservation surgery (P = 0.012). The median follow-up time was 16 months, and the estimated 16-months DFS was 98.1% for non-obese and 95.0% for obese patients (P = 0.007), the estimated 16-months OS was 99.4% for non-obese and 98.4% for obese patients (P = 0.004). The multivariate analysis indicated that obesity is an independent prognostic factor for OS and DFS in breast cancer patients.

CONCLUSIONSOur findings suggest that obesity is associated with a poorer outcome in Chinese female patients with breast cancer.

Body Mass Index ; Breast Neoplasms ; epidemiology ; China ; epidemiology ; Disease-Free Survival ; Female ; Humans ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Obesity ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Survival Analysis

Objective Obesity has been shown to be an indicator of poor prognosis for patients with primary breast cancer.The aim of this study was to clarify the effect of obesity on Chinese women with breast cancer.Methods This is a retrospective analysis of 1699 breast cancer patients.We evaluated the effect of body mass index (BMI) on disease-free survival (DFS) and overall survival (OS) in these patients.BMI was obtained before surgery.Obesity was defined as a BMI≥24.Kaplan-Meier analysis and Log rank test were employed to perform survival analysis.The impact of different characteristics on survival was assessed by using Cox proportional-hazards regression model.Results In total 635 ( 37.4%) patients were obese, while 1 064 (62.6%) were non-obese.Comparing the tumor characteristics in the two groups, the BMI≥24 group showed a higher rate of older age ( P<0.001) , postmenopausal status ( P<0.001) , increased risk of lymph node metastasis ( P=0.001) and less chances of accepting breast conservation surgery ( P=0.012) . The median follow-up time was 16 months, and the estimated 16-months DFS was 98.1%for non-obese and 95.0%for obese patients ( P=0.007) , the estimated 16-months OS was 99.4%for non-obese and 98.4%for obese patients(P=0.004).The multivariate analysis indicated that obesity is an independent prognostic factor for OS and DFS in breast cancer patients.Conclusions Our findings suggest that obesity is associated with a poorer outcome in Chinese female patients with breast cancer.