ObjectiveTo establish background data for a 90-day feeding trial of SD rats to ensure the reliability of research data. MethodsBackground data from six independent 90-day feeding trials of SD rats conducted by the National Center for Safety Evaluation of Drugs from 2020 to 2023 were summarized. These studies involved a blank control group of 120 SPF-grade 4-week-old SD rats, with an equal number of males and females, which were only given standard full-nutrient pelleted rat feed. After the quarantine period, the animals were observed for an additional 90 days, followed by intraperitoneal injection of Zoletil (50 mg/mL) for anesthesia, blood sampling, euthanasia, and necropsy. By analyzing the data from the blank control group, a relevant background database for SD rats was established. ResultsBoth male and female rats exhibited steady weight gain, with a more pronounced increase in male rats. Within 90 days, the average body weight of male and female rats increased to over 500 g and 300 g, respectively. Three weeks later, the average daily food intake of male rats stabilized at approximately 25~28 g per rat, while that of female rats remained stable at approximately 16~19 g per rat. The food utilization rate of all animals gradually decreased from the first week of the experiment. In the white blood cell (WBC) differential count results, significant differences were observed in the counts of WBCs, neutrophils (Neut), lymphocytes (Lymph), and monocytes (Mono) between males and females (P<0.001). However, there were no significant differences in the percentages of neutrophil (%Neut), lymphocyte (%Lymph), and monocyte (%Mono) between the sexes (P>0.05). The average red blood cell count (RBC), hemoglobin concentration (HGB), hematocrit (HCT), platelet count (PLT), prothrombin time (PT), and activated partial thromboplastin time (APTT) were higher in male animals than in female animals (P<0.05). The average values of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine phosphokinase (CK), lactate dehydrogenase (LDH), glucose (GLU), and triglyceride (TG) in male rats were higher than those in female rats (P<0.05). The urinary pH range for male animals was 5.0 to 8.5, while for female animals it was 6.5 to 9.0. The majority of male animals had a urinary specific gravity lower than 1.020, and the majority of female animals had a urinary specific gravity lower than 1.015. The weights of various organs (excluding the adrenal glands and reproductive organs) in male animals were heavier than those in female animals (P<0.001), while the organ/body weight ratios (excluding the kidneys and reproductive organs) of female animals were higher than those of male animals (P<0.001). ConclusionThis study summarizes the background reference ranges for body weight, food intake, hematology, and serum biochemistry indicators in SPF-grade SD rats in the untreated control group from six 90-day feeding trials conducted by the National Center for Safety Evaluation of Drugs. It provides important reference data for related research. By summarizing the background and spontaneous histopathological changes in rats, this study aids in the standardization and normalization of subsequent research, as well as in the evaluation and analysis of abnormal results.

OBJECTIVE: To observe the effects of Tongdu Tiaoshen acupuncture (for unblocking the obstruction in the governor vessel and regulating the spirit) on depression-like behavior and the hippocampal Endophilin A1/reactive oxygen species (ROS) pathway in the chronic unpredictable mild stress (CUMS) model rats, and explore the mechanism of this therapy for depression. METHODS: Forty-eight male SD rats of SPF grade were randomly divided into a normal group (n=12) and a modeling group (n=36). In the modeling group, CUMS was performed to establish depression model. The successfully-modeled rats were randomized into a model group, a Tongdu Tiaoshen acupuncture group (referred to as the acupuncture group), and a fluoxetine group, with 12 rats in each group. In the acupuncture group, "Baihui" (GV20), "Shenting" (GV24), "Shuigou" (GV26) and "Dazhui" (GV14) were stimulated with acupuncture. This intervention measure was delivered once a day, continuously for 6 days; it was discontinued on day 7 and was completed in 28 days. In the fluoxetine group, intragastric administration was done with fluoxetine solution (2.1 mg/kg), once a day, and for 28 consecutive days. Before and after modeling, and after intervention completion, the body mass, sucrose preference rate and the total distance of movement and the boxes of horizontal crossing in the open field experiment were observed in each group. After intervention, using HE staining, the hippocampal neuron morphology was observed; using Nissl staining, the hippocampal Nissl body number was counted. The hippocampal mitochondria was observed under transmission electron microscopy. The average fluorescence intensity of ROS in hippocampal was determined using flow cytometry. With Western blot method, the protein expression of Endophilin A1, growth associated protein 43 (GAP-43), and brain-derived neurotrophic factor (BDNF) in hippocampal was detected; and with RT-qPCR method, the mRNA expression of Endophilin A1, GAP-43, and BDNF was recorded. Using the immunofluorescence, the average fluorescence intensity of Endophilin A1, GAP-43, and BDNF in hippocampal tissue was determined. RESULTS: Compared with the normal group, in the model group, the body mass, sucrose preference rate, and the total distance of movement and the boxes of horizontal crossing in the open field experiment decreased (P<0.01); the hippocampal neuronal structure was unclear, the matrix was relatively loose, and the number of Nissl body decreased (P<0.01); mitochondrial structure was disarranged, the outer membrane was ruptured, mitochondrial cristae was irregular or missed; the average fluorescence intensity of ROS in hippocampal tissue, the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue increased (P<0.01), while the protein and mRNA expression of GAP-43 and BDNF and its average fluorescence intensity decreased (P<0.01). Compared with the model group, the acupuncture group and the fluoxetine group showed the increase in body mass, sucrose preference rate, the total distance of movement and the boxes of horizontal crossing in the open field experiment (P<0.05, P<0.01); the hippocampal neuronal structure became relatively clear, the matrix was relatively dense, and the number of Nissl body was elevated (P<0.01); mitochondrial structure got normal and disarranged slightly, the average fluorescence intensity of ROS in hippocampal tissue, the protein and mRNA expression and the average fluorescence intensity of Endophilin A1 in hippocampal tissue were reduced (P<0.01), while the protein and mRNA expression of GAP-43 and BDNF and the average fluorescence intensity rose (P<0.01, P<0.05). Compared with the fluoxetine group, the acupuncture group presented the increase in the average fluorescence intensity of ROS, the protein expression and the average fluorescence intensity of Endophilin A1, the protein expression of GAP-43 and the mRNA expression of BDNF (P<0.01, P<0.05), and the decrease of the protein expression and the average fluorescence intensity of BDNF, the mRNA expression of Endophilin A1, and the average fluorescence intensity of GAP-43 (P<0.01, P<0.05). CONCLUSION Tongdu tiaoshen acupuncture alleviates depression-like behaviors in CUMS model rats and protects hippocampal neurons, which may be related to suppressing Endophilin A1/ROS signaling pathway and attenuating oxidative stress reactions.
Animals ; Male ; Hippocampus/metabolism* ; Acupuncture Therapy ; Rats, Sprague-Dawley ; Rats ; Depression/psychology* ; Humans ; Reactive Oxygen Species/metabolism* ; Disease Models, Animal ; Acupuncture Points

Hypertrophic scar is a fibrous hyperplastic disorder that arises from skin injuries. The current therapeutic modalities are constrained by the dense and rigid scar tissue which impedes effective drug delivery. Additionally, insufficient autophagic activity in fibroblasts hinders their apoptosis, leading to excessive matrix deposition. Here, we developed an active microneedle (MN) system to overcome these challenges by integrating micromotor-driven drug delivery with autophagy regulation to remodel the scar microenvironment. Specifically, sodium bicarbonate and citric acid were introduced into the MNs as a built-in engine to generate CO₂ bubbles, thereby enabling enhanced lateral and vertical drug diffusion into dense scar tissue. The system concurrently encapsulated curcumin (Cur), an autophagy activator, and triamcinolone acetonide (TA), synergistically inducing fibroblast apoptosis by upregulating autophagic activity. In vitro studies demonstrated that active MNs achieved efficient drug penetration within isolated scar tissue. The rabbit hypertrophic scar model revealed that TA-Cur MNs significantly reduced the scar elevation index, suppressed collagen I and transforming growth factor-β1 (TGF-β1) expression, and elevated LC3 protein levels. These findings highlight the potential of the active MN system as an efficacious platform for autonomous augmented drug delivery and autophagy-targeted therapy in fibrotic disorder treatments.

Vascular damage plays a significant role in the onset and progression of Alzheimer's disease (AD). However, the precise molecular mechanisms underlying the induction of neuronal injury by vascular damage remain unclear. The present study aimed to examine the impact of fibrinogen (Fg) on tau pathology. The results showed that Fg deposits in the brains of tau P301S transgenic mice interact with tau, enhancing the cytotoxicity of pathological tau aggregates and promoting tau phosphorylation and aggregation. Notably, Fg-modified tau fibrils caused enhanced neuronal apoptosis and synaptic damage compared to unmodified fibrils. Furthermore, intrahippocampal injection of Fg-modified tau fibrils worsened the tau pathology, neuroinflammation, synaptic damage, neuronal apoptosis, and cognitive dysfunction in tau P301S mice compared to controls. The present study provides compelling evidence linking Fg and tau, thereby connecting cerebrovascular damage to tau pathology in AD. Consequently, inhibiting Fg-mediated tau pathology could potentially impede the progression of AD.
Animals ; tau Proteins/metabolism* ; Alzheimer Disease/metabolism* ; Fibrinogen/metabolism* ; Mice, Transgenic ; Mice ; Disease Models, Animal ; Memory Disorders/metabolism* ; Male ; Mice, Inbred C57BL ; Brain/metabolism* ; Hippocampus/metabolism* ; Protein Aggregation, Pathological/metabolism* ; Apoptosis ; Phosphorylation

Metabolic dysfunction-associated fatty liver disease (MAFLD), characterized by fatty acid overload, secondary chronic inflammation, and fibrosis, has become the most prevalent chronic liver disease globally. While no effective pharmacotherapy exists for MAFLD, mitigating inflammatory responses represents a promising approach to preventing the progression from steatosis to severe steatohepatitis. The NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, which detects endogenous danger and stress signals, has emerged as a significant target for inflammatory disease treatment, as transcriptional inactivation of its components demonstrates the therapeutic potential for MAFLD. Natural products targeting NLRP3 inflammasome activation have shown promising efficacy in MAFLD therapy. This review synthesizes the current understanding of NLRP3 inflammasome activation and therapeutic targets for NLRP3 homeostasis. Additionally, natural products reported to inhibit NLRP3 inflammasome for MAFLD improvement are categorized according to their mechanisms of action. The review also addresses limitations and future directions regarding natural products targeting NLRP3 inflammasome in MAFLD treatment. Enhanced understanding of NLRP3 inflammasome activation mechanisms in MAFLD and the identification of novel natural products supported by mechanistic research will significantly advance MAFLD treatment.
Humans ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology* ; Inflammasomes/metabolism* ; Biological Products/therapeutic use* ; Animals ; Fatty Liver/immunology*

The pathogenesis and progression of myocardial fibrosis(MF)are modulated by diverse cytokines and signaling pathways.Studies in recent years have revealed an extremely close association between MF evolution and programmed cell death(PCD)modali-ties,including apoptosis,autophagy,pyroptosis,necrosis,ferroptosis,and cuproptosis.This article reviews the characteristics of MF and distinct PCD modalities,with particular emphasis on cutting-edge discoveries elucidating PCD in the field of MF,hoping to pro-vide a theoretical-level reference basis for the in-depth exploration and clinical intervention of MF.

Objective To investigate the differences in acute mountain sickness between individuals en-tering the plateau for the first time and those with multiple plateau experiences,and to examine the impact on psychological stress.Methods A total of 2 337 young plateau-acclimatized individuals from an acclimatization rotation area during November 2023 to January 2024 were selected as the study subjects.Those entering the plateau for the first time were classified into Group A(n=425),while those with previous plateau experience(at least one prior visit)were classified into Group B(n=1 912).Differences were analyzed between the two groups in terms of general characteristics,as well as acute mountain sickness(AMS)scores,Self-Rating De-pression Scale(SDS)scores,Symptom Checklist-90(SCL-90)scores,and various vital signs at the following time points:before entering the plateau(T0),on the 1st day(T1),4th day(T2),7th day(T3),and 10th day(T4)after entering the plateau.Results Group A was younger than Group B,with a statistically significant difference(P<0.05).At T1,T2,and T3,AMS scores in Group A were significantly higher than those in Group B(P<0.05).At T4,there was no statistically significant difference in AMS scores between the two groups(P>0.05).At T1,T2,and T3,blood oxygen saturation was lower and heart rate higher in Group A than in Group B,with statistically significant differences(P<0.05).After high-altitude acclimatization,no statistically significant differences in blood oxygen saturation or heart rate were observed between the groups at T4(P>0.05).At T1,there were no statistically significant differences in systolic or diastolic blood pres-sure between the groups(P>0.05).At T2,both systolic and diastolic blood pressure were higher in Group A than in Group B(P<0.05).At T3 and T4,after high-altitude acclimatization,systolic blood pressure de-creased in both groups but remained higher in Group A than in Group B(P<0.05).On the SDS scale,men-tal-emotional and somatic symptom scores were significantly higher in Group A than in Group B(P<0.001).On the SCL-90,all nine symptom factor scores were higher in Group A than in Group B(P<0.05).Conclusion Indi-viduals entering high-altitude areas for the first time experience more severe acute mountain sickness and more pronounced psychological stress.Therefore,enhanced monitoring,psychological counseling,and tailored pre-vention and treatment strategies should be implemented.

Objective To investigate the relationship between the levels of type Ⅲ procollagen(PCⅢ)and heme-binding protein(HPX)in peripheral blood,blood lipid and liver fibrosis in patients with nonalcoholic steatohepatitis(NASH).Methods Totally 126 patients with NASH admitted to the hospital from January 2021 to December 2023 were selected as NASH group,and they were divided into 3 subgroups according to the degree of liver fibrosis:no liver fibrosis group(22 cases),mild group(63 cases),moderate and severe group(41 cases).In addition,150 healthy subjects in the same period were selected as the control group.Enzyme-linked immunosorbent assay(ELISA)was used to detect and compare the levels of PC Ⅲ and HPX in periph-eral blood of all groups,and Pearson correlation analysis was used to explore the correlation between peripher-al blood PC Ⅲ and HPX levels and related indexes of blood lipids in NASH patients.The predictive value of peripheral blood PC Ⅲ and HPX for liver fibrosis in NASH patients was evaluated by receiver operating char-acteristic(ROC)curve.Results The levels of PC Ⅲ and HPX,as well as total cholesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)in NASH group were higher than those in control group,and the levels of high density lipoprotein cholesterol(HDL-C)were lower than those in control group,and the differences were statistically significant(P＜0.05).Pearson correlation analysis showed that the lev-els of PCⅢ and HPX in peripheral blood of NASH patients were positively correlated with TC,TG and LDL-C(P＜0.05),while negatively correlated with HDL-C(P＜0.05).The levels of PC Ⅲ and HPX in peripheral blood of the non-fibrosis group were lower than those of the mild and moderate severe groups,and the mild group was lower than that of the moderate and severe group,and the differences were statistically significant(P＜0.05).The area under the curve(AUC)of peripheral blood PC Ⅲ and HPX in predicting liver fibrosis in NASH patients were 0.757 and 0.861,respectively,and the cut-off values were 135.51 ng/mL and 804.86 mg/L,respectively.The AUC of the combined prediction for liver fibrosis was 0.905,which was higher than that predicted by the two alone.Conclusion The level of PC Ⅲ and HPX in peripheral blood is closely related to the level of blood lipids and the degree of liver fibrosis in NASH patients,which can be used as potential bi-omarkers to predict liver fibrosis in NASH patients.

Protein polypeptides are a class of bioactive substances that play a crucial role in maintaining the stability of various functions of the organism.Rapid and accurate detection of their levels could help in the diagnosis of diseases,the monitoring of drug therapy and the research and development of medicines,which is of great significance in the fields of clinical medicine,biology and pharmacy.Conventional protein polypeptides detection methods,such as Western blotting and enzyme-linked immunosorbent assay,still have problems like low sensitivity or difficulty in determining more than 1 analyte simultaneously.Liquid chromatography-mass spectrometry(LC-MS)has the advantages of specificity,sensitivity and high throughput.However,low ionization efficiency of macromolecules and strong biological matrix effects limit the feasibility of direct detection of protein polypeptides by LC-MS,which has led to the development of signal conversion and amplification strategies based on LC-MS technology.In this review,we summarized the research progress of sample pretreatment methods and signal conversion and amplification strategies for quantification of protein polypeptides in vivo based on LC-MS in recent years,providing a reference for developing specific high-sensitivity detection methods for low-abundance protein polypeptides in complex samples based on LC-MS technology.

Objective To explore the diagnostic value of alkaline phosphatase on the surface membrane of neutrophils(mNAP)in bloodstream infections(BSI)of Gram-negative bacteria(GNB)and Gram-positive bacteria(GPB).Methods A total of 418 patients diagnosed with BSI at Donghai People's Hospital from January 2022 to December 2023 were enrolled in the study.Based on the results of Gram staining from positive blood cultures,the patients were divided into GNB bacteremia(n=329)and GPB bacteremia(n=89).Additionally,35 hospitalized patients with systemic inflammatory response syndrome(SIRS)during the same period were selected as the control group.Their clinical data,routine laboratory test results,blood cultures and venous blood samples were collected,and the expression levels of mNAP were detected by flow cytometry.The receiver operating characteristics(ROC)curve was used to evaluate the diagnostic performance of mNAP for BSI of GNB and GPB.Results The expression levels of mNAP in the GPB infection,GNB infection and SIRS groups were 9 588(5 677,11 343)AB/C,16 616(11 853,22 035)AB/C,and 5 738(2 613,9 178)AB/C,respectively,and the difference among them was statistically significant(H=43.95,P＜0.000 1).Further pairwise comparisons showed that the expression levels of mNAP in the GNB infection group were significantly higher than those in the GPB infection group(U=203.0,P＜0.000 1)and the SIRS group(U=445.0,P＜0.000 1).Meanwhile,the expression levels of mNAP in the GPB infection group were significantly higher than that in the SIRS group(U=583.0,P＜0.000 1).The area under the ROC curve(AUCROC)of mNAP for predicting the BSI of GNB was 0.91(95%CI:0.85-0.96).When the cut-off value was 10 820 AB/C,its sensitivity and spe-cificity were 80.00%and 88.57%,respectively.The AUCROC of mNAP for predicting the BSI of GPB was 0.69(95%CI:0.55-0.83).When the cut-off value was 10 859 AB/C,its sensitivity and specificity were 33.00%and 88.13%,respectively.Conclusion The di-agnostic efficiency of mNAP for the BSI of GNB is significantly higher than that for the BSI of GPB,which may become a novel biologi-cal marker for distinguishing the BSI of GNB and GPB.