OBJECTIVE To establish a method for determining the content of 11 antioxidants and their degradation products in recombinant Exendin-4-FC fusion protein injection by HPLC. METHODS The protein was precipitated with saturated ammonium sulfate. After centrifugation, the supernatant was transferred to a C18 solid phase extraction cartridge activated by methanol. Then the cartridge was eluted with 4 mL of methanol and 5 mL of ethyl acetate respectively, and the eluent was diluted with methanol-ethyl acetate(2∶3) mixed solvent and passed through a 0.22 µm PTFE hydrophobic filter. It was analyzed by HPLC and quantified by external standard method. Chromatographic conditions: Kinetex® XB-C18 100Å (100 mm×4.6 mm, 2.6 µm)column, the detection wavelength was 230 nm, the column oven was 30 ℃, the injection volume was 5 µL and the flow rate was 0.4 mL·min–1, mobile phase was 0.1% formic acid-methanol(A)-0.1% formic acid aqueous solution(B), the running time was 45 min. RESULTS The 11 target substances showed a good linear relationship in the range of 2.5−35 μg·mL–1 with R2 ≥0.99. At three different concentration(25, 10, 5 μg·mL–1) of spiked samples, the average recovery rates of 11 antioxidants ranged from 88.1% to 106.5%, with RSDs in the range of 0.10%–9.05%. The RSDs of 6 repeatable samples was 2.01%–4.77%, which of 12 intermediate precision samples was 2.58%–9.75%. The positive/inverted samples of three batches of recombinant Exendin-4-FC fusion protein injection were detected at 0 month, 3 months and 6 months(25 ℃), and the results showed that there was no antioxidant and its degradation leaching in all batches of samples at different detection points. CONCLUSION The method has good specificity, high accuracy and precision, good solution stability, high durability and can be used for the content detection of antioxidants in drugs.

A high-altitude environment is characterized by low oxygen levels,low pressure,and low temperatures.Exposure to the plateau environment often causes damage to the body,leading to the occurrence of acute mountain sickness/chronic mountain sickness(AMS/CMS).Research indicates that acute or chronic exposure to the special environment can result in overall organ dysfunctions,such as those in the heart and gastrointestinal tract.The damage to the body caused by exposure to the plateau environment is closely related to acute and chronic hypoxia.Physiological maladjustment or disease is usually accompanied by changes in the structure of the gut microbiota.There have been reports on the correlations between the gut microbiota and bodily harm caused by high-altitude exposure.However,the specific types of bacteria involved and the mechanisms of action are still under investigation.This article reviews the intestinal tissue damage caused by low oxygen levels,immune activation,changes in microbial community structure,and differential metabolic products.The association and underlying mechanisms between bodily harm due to high-altitude exposure and the intestinal microbiota are also explored in hopes of stimulating new lines of thought related to the prevention and treatment of bodily harm caused by exposure to the plateau environment.

Objective To explore the effectiveness of high-definition diffusion weighted imaging(DWI)sequence combined with T1 WI-fat suppression(FS)dynamic contrast enhancement(DCE)sequence for preoperative T-stage of rectal cancer by using 3.0T MRI standardized scanning.Methods A retrospective analysis was conducted on MRI images of 57 patients with rectal cancer confirmed by pathology.Before surgery,the patients underwent 3.0T MRI standardized rectal cancer scan methods,including routine sequence,high-definition DWI sequence,and T1 WI-FS DCE sequence,etc.Then two experienced physicians evaluated the T-stage of preoperative rectal cancer through high-definition DWI(transverse and sagittal sections)and T1 WI-FS DCE sequences in the double-blind method.Using the postoperative pathological results of rectal cancer as the"gold standard",two sequences were combined to evaluate the accuracy,sensitivity,and specificity of rectal cancer T-stage.Results Among the 57 cases,there were 9 cases of upper rectal cancer,39 cases of middle rectal cancer,and 9 cases of lower rectal cancer.The accuracy rates of preoperative T-stage diagnosis for rectal cancer by two evaluator were both 85.7％(6/7)in T1 stage,88.2％(15/17)and 94.1％(16/17)in T2 stage,96.9％(31/32)and 93.8％(30/32)in T3 stage,and both 100.0％(1/1)in T4 stage.For evaluator 1,the sensitivity and specificity of the rectal cancer T-stage diagnosis were 96.1％and 83.3％,and for evaluator 2 were 94.1％and 83.3％,respectively.For rectal cancer MRI diagnosis,the accuracy rates and sensitivity were higher when combining the high-definition DWI sequence and T1 WI-FS DCE sequence,compared with a single high-definition DWI sequence or T1 WI-FS DCE sequence,and the difference was statistically significant.The average preoperative apparent diffusion coefficient(ADC)value of rectal cancer was compared between the corresponding postoperative pathological T1 to T4 stage groups,and the difference was statistically significant.Conclusion The combination of high-definition DWI sequence and T1 WI-FS DCE sequence improves the accuracy of rectal cancer T-stage,providing assistance for personalized clinical treatment.

Objective:To explore the influence of deviation of the bolt in femoral neck system (FNS) on the short-term outcomes in young and middle-aged patients with displaced femoral neck fracture.Methods:A retrospective analysis was conducted of the 114 young and middle-aged patients with displaced femoral neck fracture who had been treated with FNS at Department of Orthopaedics, Suzhou Municipal Hospital from December 2019 to January 2023. Based on the postoperative measurements of the deviation of the bolt tip to the central axis of the femoral head and neck (W), the patients were divided into a central group (W≤20%) and a deviation group (W>20%). In the central group of 63 cases, there were 27 males and 36 females with a mean age of (46.4±8.0) years. In the deviation group of 51 cases, there were 20 males and 31 females with a mean age of (45.1±9.8) years. The 2 groups were compared in terms of weight-bearing time, fracture healing time, tip-apex distance, degree of femoral neck shortening, Harris Hip Score and EuroQol five-dimensional questionnaire-5L (EQ-5D-5L) utility value at the last follow-up, as well as complications and revision surgeries.Results:There was no statistically significant difference in the preoperative general information, auxiliary reduction or quality of fracture reduction between the 2 groups, showing comparability between groups ( P>0.05). There was no significant difference in the partial weight-bearing time between the 2 groups ( P>0.05). In the central group, the full weight-bearing time [15.0 (14.0, 16.0) weeks] and fracture healing time [14.0 (12.0, 15.0) weeks] were significantly shorter than those in the deviation group [16.0 (15.0, 19.0) weeks; 15.0 (13.0, 17.0) weeks], the tip-apex distance [(21.4±3.4) mm] was significantly shorter than that in the deviation group [(23.5±2.7) mm], the Harris Hip Score [(90.6±6.1) points] and EQ-5D-5L utility value [0.9 (0.8, 0.9)] at the last follow-up were significantly higher than those in the deviation group [(87.7±6.2) points; 0.9 (0.8, 0.9)], and the incidences of moderate and severe femoral neck shortening [25.4% (16/63)], avascular necrosis of the femoral head [0 (0/63)] and revision surgery [0 (0/63)] were significantly lower than those in the deviation group [66.7% (34/51), 7.8% (4/51), 9.8% (5/51)] (all P< 0.05). Conclusion:A closer positioning of the FNS bolt to the central axis of the femoral head and neck favors satisfactory short-term outcomes and a lower revision surgery rate in young and middle-aged patients with displaced femoral neck fracture.

Stroke is a cerebrovascular disease characterized by high rates of disability,recurrence and mortal-ity.It results primarily from cerebrovascular rupture or transient/permanent occlusion.Due to the complex pathological mechanism of stroke and the lack of early-stage biomarkers,effective treatment options remain limited.Circular RNAs(circRNAs)are stable and conserved endogenous long noncoding RNA that regulate the pathological process of stroke,in-cluding oxidative stress,inflammation,apoptosis and autophagy.In addition,the regulatory roles of circRNAs in stroke include ceRNA,DNA methylation and histone modification.This review provides a comprehensive overview of circRNA biogenesis,major functions and characteristics.Furthermore,it presents the latest research findings on circRNAs in the context of stroke pathology,with the aim of providing new insights and potential approaches for unraveling the underlying mechanism,diagnosis and treatment of such diseases.

Background::Hypothermia therapy has been suggested to attenuate myocardial necrosis; however, the clinical implementation as a valid therapeutic strategy has failed, and new approaches are needed to translate into clinical applications. This study aimed to assess the feasibility, safety, and efficacy of a novel selective intracoronary hypothermia (SICH) device in mitigating myocardial reperfusion injury.Methods::This study comprised two phases. The first phase of the SICH was performed in a normal porcine model for 30 minutes ( n = 5) to evaluate its feasibility. The second phase was conducted in a porcine myocardial infarction (MI) model of myocardial ischemia/reperfusion which was performed by balloon occlusion of the left anterior descending coronary artery for 60 minutes and maintained for 42 days. Pigs in the hypothermia group ( n = 8) received hypothermia intervention onset reperfusion for 30 minutes and controls ( n = 8) received no intervention. All animals were followed for 42 days. Cardiac magnetic resonance analysis (five and 42 days post-MI) and a series of biomarkers/histological studies were performed. Results::The average time to lower temperatures to a steady state was 4.8 ± 0.8 s. SICH had no impact on blood pressure or heart rate and was safely performed without complications by using a 3.9 F catheter. Interleukin-6 (IL-6), tumor necrosis factor-α, C-reactive protein (CRP), and brain natriuretic peptide (BNP) were lower at 60 min post perfusion in pigs that underwent SICH as compared with the control group. On day 5 post MI/R, edema, intramyocardial hemorrhage, and microvascular obstruction were reduced in the hypothermia group. On day 42 post MI/R, the infarct size, IL-6, CRP, BNP, and matrix metalloproteinase-9 were reduced, and the ejection fraction was improved in pigs that underwent SICH.Conclusions::The SICH device safely and effectively reduced the infarct size and improved heart function in a pig model of MI/R. These beneficial effects indicate the clinical potential of SICH for treatment of myocardial reperfusion injury.

In order to explore the effects of Coptis detoxification powder on immune suppression and inflammatory response caused by heat stress in animals,Bama pigs were used as experimental animals to establish an animal model of heat stress,and the effects of Coptis detoxification powder on transcriptional activation of TLRs mRNA in PBMC induced by heat stress and its mediated ex-pression of inflammatory factors were studied in vivo.The animal model of heat stress was estab-lished by artificial climate greenhouse,Chinese medicine"Coptis detoxification powder"was pre-pared,and its inhibitory effect on heat stress and inflammation was studied by preventive and therapeutic administration.Real-time fluorescence quantitative PCR was used to determine the transcriptional changes of HSP70,TLRs and IL-17 mRNA,and ELISA was used to determine the dynamic changes of serum IL-2,IL-6 and TNF-α.Western blot and indirect immunofluorescence(IFA)were used to determine the nuclear translocation and protein expression of p65 phosphoryl-ated protein in NF-κβ inflammatory factor pathway.The transcription of PBMC TLRs mRNA and its mediated inflammatory factor expression and function in Bama pigs under heat stress were sys-tematically analyzed.The results showed that the experimental animal model of Bama pigs under heat stress was successfully established,and heat stress significantly up-regulated the mRNA tran-scription levels of HSP70,TLR2,TLR4 and inflammatory factor IL-17,resulting in significantly enhanced expressions of serum inflammatory factors IL-2,IL-6 and TNF-α.The NF-κβ pathway significantly promoted the level of p65 nuclear phosphorylation and the expression of cytoplasmic protein.The Chinese medicine"Coptis detoxification powder"could significantly reduce the upreg-ulation effect of heat stress conditions on the transcription levels of HSP70 and TLRs mRNA,thus inhibiting the mRNA transcription of inflammatory factor IL-17 and the expression of IL-2,IL-6 and TNF-α in serum,and significantly antagonizing the transcriptional activity of NF-κβ caused by heat stress.It showed good anti-heat stress effect.The anti-inflammatory effects of Chinese veteri-nary medicine prescription"Coptis detoxification powder",its pathway of action and its effectors were investigated,the expression and secretion levels of the regulatory factors related to the path-way associated with TLRs mRNA inhibition were analyzed,and the functional effects and mecha-nism of its clinical anti-heat stress and anti-inflammatory effects were clarified,which laid a data foundation for the promotion and application of this prescription and related mechanism research.

A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.
Animals ; Mice ; Peste-des-Petits-Ruminants/prevention & control* ; Antibodies, Monoclonal ; Reproducibility of Results ; Peste-des-petits-ruminants virus ; Antibodies, Viral ; Enzyme-Linked Immunosorbent Assay ; Goats

OBJECTIVE: To investigate the relationship between malnutrition and delirium and its effect on prognosis in elderly patients with severe pneumonia undergoing invasive mechanical ventilation. METHODS: A prospective observational study was conducted. Patients with severe pneumonia aged ≥ 60 years old who underwent invasive mechanical ventilation admitted to department of critical care medicine of the Second People's Hospital of Lianyungang from January 2021 to December 2022 were enrolled. The confusion assessment method (CAM) was used to evaluate the delirium of the patients in intensive care unit (ICU). The score of CAM ≥ 1 was defined as delirium. Mini nutritional assessment short-form (MNA-SF) was used to assess the nutritional status of patients, and MNA-SF score ≤ 7 was defined as malnutrition. Patients were divided into delirium group and non-delirium group according to whether delirium occurred. The differences in clinical indicators, length of ICU stay, duration of mechanical ventilation and wake-up time after drug withdrawal were compared between the two groups. After 28 days of short-term follow-up, the patients were divided into death group and survival group, and the differences in the incidence of delirium and malnutrition between the two groups were compared. Binary multivariate Logistic regression analysis was used to screen the risk factors for delirium in elderly patients with severe pneumonia undergoing invasive mechanical ventilation. Kaplan-Meier survival curve was used to analyze the effect of delirium on prognosis. RESULTS: A total of 132 elderly patients with severe pneumonia undergoing invasive mechanical ventilation were enrolled, of whom 98 survived and 34 died within 28 days, with a mortality of 25.76%. The incidence of malnutrition and delirium in the death group was significantly higher than that in the survival group (61.76% vs. 37.76%, 64.71% vs. 26.53%, both P < 0.05), and the MNA-SF score was significantly lower than that in the survival group (6.32±1.80 vs. 8.72±2.23, P < 0.01). Procalcitonin (PCT), interleukin-6 (IL-6) and blood lactic acid (Lac) in the death group were significantly higher than those in the survival group [PCT (μg/L): 4.47 (2.69, 10.39) vs. 2.77 (1.28, 5.94), IL-6 (ng/L): 204.08 (126.12, 509.85) vs. 120.46 (60.67, 290.99), Lac (mmol/L): 5.14 (2.75, 8.60) vs. 3.13 (2.16, 4.30), all P < 0.05], and the wake-up time after drug withdrawal was significantly longer than that in the survival group (minutes: 33.94±8.51 vs. 28.92±7.03, P < 0.01). Among 132 elderly patients with severe pneumonia undergoing invasive mechanical ventilation, 48 patients had delirium during ICU stay, and 84 patients did not have delirium. The incidence of delirium was 36.36%. The 28-day mortality in the delirium group was significantly higher than that in the non-delirium group (45.83% vs. 14.29%, P < 0.01), and the MNA-SF score was significantly lower than that in the non-delirium group (6.46±1.77 vs. 9.05±2.15, P < 0.01), the length of ICU stay, duration of mechanical ventilation, and wake-up time after drug withdrawal were also significantly longer than those in the non-delirium group [length of ICU stay (days): 13.40±9.59 vs. 10.06±7.81, duration of mechanical ventilation (hours): 197.06±89.80 vs. 138.81±82.30, wake-up time after drug withdrawal (minutes): 35.85±7.01 vs. 26.99±6.12, all P < 0.05]. Binary multivariate Logistic regression analysis showed that malnutrition [odds ratio (OR) = 7.527, 95% confidence interval (95%CI) was 2.585-21.917], Lac (OR = 5.345, 95%CI was 1.733-16.483), wake-up time after drug withdrawal (OR = 6.653, 95%CI was 2.021-21.904) were independent risk factors for delirium during ICU stay in elderly patients with severe pneumonia undergoing invasive mechanical ventilation (all P < 0.01). Kaplan-Meier survival analysis showed that the 28-day cumulative survival rate of patients in the delirium group was significantly lower than that in the non-delirium group (54.17% vs. 85.71%), and the difference was statistically significant (Log-Rank test: χ² = 16.780, P < 0.001). CONCLUSIONS The risk factors for delirium in elderly patients with severe pneumonia undergoing invasive mechanical ventilation during ICU stay include malnutrition, Lac, and wake-up time after drug withdrawal. The occurrence of delirium is closely related to poor prognosis.
Aged ; Humans ; Middle Aged ; Respiration, Artificial ; Interleukin-6 ; Pneumonia ; Intensive Care Units ; Delirium/etiology* ; Procalcitonin ; Prognosis ; Malnutrition ; Retrospective Studies

Alzheimer's disease （AD） is a neurodegenerative disorder characterized by mechanisms including excessive deposition of β amyloid （Aβ）， neuroinflammatory responses， and hyperphosphorylation of microtubule-associated protein （Tau protein）. Currently， there are no effective clinical treatments available for AD. Traditional Chinese herbal drugs have gained attention for their potential to exert anti-AD effects through multi-component， multi-target approaches. As a traditional Chinese herbal drug with over 600 years of clinical use， Geranii Herba has substantial medicinal potential and wide application prospects. Geranium medicinal plants contain chemical components such as tannins， flavonoids， organic acids， and volatile oils. Research has indicated that various tannin compounds found in Geranii Herba possess pharmacological activities like enhancing learning and memory abilities， and improving cognitive function. These effects are linked to mechanisms involving anti-Aβ effects， Tau protein regulation， antioxidation， anti-inflammation， and inhibition of acetylcholinesterase activity. These compounds can act through multiple pathways and targets to inhibit neurodegenerative changes in neurons， thus effectively preventing and treating neurodegenerative diseases like AD. Based on relevant literature， this study focused on reviewing various tannin components in Geranium plants and their role in preventing and treating AD and identified potential drug components for treating neurodegenerative diseases such as AD by exploring the tannin components and their mechanisms in Geranium plants， thereby providing a theoretical foundation and research direction for further development and clinical application.