ObjectiveTo investigate the inhibitory effect of Huoluo Xiaolingdan on triple negative breast cancer （TNBC） in mice through its regulation of TCF1⁺CD8⁺ stem cell-like T cells infiltration. MethodsA mouse model of TNBC was established and the mice were randomly divided into the model group， low-dose （3.9 g·kg^-1）， medium-dose （7.8 g·kg^-1） and high-dose （15.6 g·kg^-1） Huoluo Xiaolingdan groups， and anti-PD-1 antibody treatment group. Each group was given a dose of 0.01 mL·g^-1， while the model group and the anti-PD-1 treatment group were also given an equivalent volume of normal saline. The drug was administered for 21 days. In the anti-PD-1 antibody group， mice were intraperitoneally injected with 100 μg of mouse anti-PD-1 antibody twice a week， for a total of five injections. The tumor volume， survival time and tumor mass were measured at different time points. Hematoxylin-eosin （HE） staining was used to observe the histological changes of the tumor. The expression of CD8⁺T cells and TCF1⁺CD8⁺ stem-like T cells in tumor tissues was detected by immunohistochemistry and immunofluorescence staining. Flow cytometry was used to detect the difference of immune cell subsets in tumors and the expression difference of TCF1⁺CD8⁺ stem cell-like T cells in tumors and peripheral blood. The expression level of PD-L1 in tumor tissues was detected by Western blot. ResultsCompared with model group， the tumor volume and mass of in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group were decreased （P<0.05， P<0.01）. The median survival time of mice in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group was as follows： 27.00 days （95%CI， 0.45-2.65）， 31.00 days （95%CI， 0.32-1.89）， 34.00 days （95%CI， 0.40-2.33）， and 35.00 days （95%CI， 0.42-2.47）. All of them were higher than that of the model group ［24.50 days （95%CI， 0.37-10.5）］. Flow cytometry showed that compared with the model group， the proportion and number of infiltrating CD8⁺ T cells in tumor were increased in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group （P<0.05， P<0.01）， while the proportion of tumor regulatory T cells （Treg） and M2 macrophages decreased （P<0.05）. Compared with the model group， the proportion of IFN-γ⁺CD8⁺ T and GrzB⁺CD8⁺ T cells in tumors in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group was increased （P<0.01）， and the proportion of TCF1⁺CD8⁺ T cells in tumor and peripheral blood was also increased. Immunofluorescence staining further showed that the number of TCF1⁺CD8⁺ T cells in tumor tissues increased in low-， medium- and high-dose Huoluo Xiaolingdan groups. Western blot analysis showed no significant decrease in the PD-L1 protein expression in tumor tissues between the Huoluo Xiaolingdan groups and the model group. ConclusionHuoluo Xiaolingdan can inhibit TNBC in mice by increasing tumor infiltration of TCF1⁺CD8⁺ stem-like T cells， enhancing CD8⁺ T cell activity， and regulating immune cell subgroups such as M2 macrophages and Treg cells to enhance anti-tumor immunity. This study provides a theoretical basis for the clinical application of Huoluo Xiaolingdan in breast cancer treatment and combination therapy.

Avian coccidiosis, an acute parasitic disease that mainly harms chicks, is widely prevalent across the world, which poses a serious threat to poultry industry. Because of the single prophylactic formulations, veterinary clinical treatment of coccidiosis mainly relies on chemically synthesized agents, polyether ionophores and Chinese herbal medicines. The introduction of novel anticoccidial drugs is slow for a long period of time, and there is an increasing problem of anticoccidial drug resistance following long-term use, which has become an urgent problem to be solved in poultry industry. This review summarizes the levels of anticoccidial drug resistance across China from 2018 to 2023, and analyzes the resistance to various anticoccidial agents in coccidia. It is indicated that the overall prevalence of anticoccidial drug resistance is high in coccidia, and development of novel anticoccidial agents and products with reduced antibiotics use and alternatives of antibiotics is of an urgent need.

Background Existing studies suggest that cadmium exposure is associated with the occurrence of cardiovascular disease (CVD), but the current epidemiological evidence is inconsistent. Objective To systematically evaluate the relationship between cadmium exposure and CVD through meta-analysis. Methods We systematically searched PubMed, Cochrane Library, Web of Science, CNKI, Wanfang and SinoMed databases to collect observational studies on the relationship between cadmium exposure and CVD in human population published until July 30, 2024. On the basis of following predetermined inclusion and exclusion criteria, the retrieved literature was systematically screened, and the basic information of the included research was extracted, including basic participant information, research outcomes, and data results. This study used the Newcastle-Ottawa Scale and a cross-sectional study evaluation protocol (11 items) recommended by the Quality of Health Care and Research Institutions of the United States for literature quality evaluation. Meta-analysis, subgroup analysis, sensitivity analysis, and assessment of publication bias of the data were conducted using Stata16.0 software. Result A total of 15 studies (18 datasets) were included, and the quality of all the studies was graded as medium or above. Among them, there were 10593 cases of CVD and 86801 controls. The results of meta-analysis showed that the standard mean difference (SMD) of the CVD case group was 0.44 and 95% confidence interval (95%CI) was (0.30, 0.58), and the cadmium exposure level of the CVD case group was higher than that of the control group (P<0.05). The subgroup analysis showed that the cadmium exposure levels in the CVD patients from the Americas (SMD=0.46, 95%CI: 0.29, 0.63) and Europe (SMD=0.14, 95%CI: 0.09, 0.19) were higher than those in the control group (P<0.05). The cadmium levels in blood (SMD=0.52, 95%CI: 0.31, 0.73) and urine (SMD=0.34, 95%CI: 0.16, 0.52) in the CVD case group were higher than those in the control group (P<0.05). In those cross-sectional studies (SMD=0.34, 95%CI: 0.23, 0.46), the cadmium exposure level in the CVD patients was higher than that in the control group (P<0.05). The sensitivity analysis showed robust meta-analysis results and the assessment of publication bias also showed no publication bias. Conclusion Cadmium exposure may increase the risk of cardiovascular disease after multiple studies were combined by meta analysis, suggesting that the scope and level of cadmium exposure in the population should be further controlled.

With the accelerating process of population aging, binocular vision disorders have become increasingly prevalent, posing new challenges for both social aging strategies and ophthalmic clinical practice. Non-strabismic binocular vision anomalies(NSBVAs)are notably prevalent among elderly populations; however, current research predominantly focuses on the abnormality rates derived from individual binocular vision clinical tests, while data on the actual prevalence based on comprehensive diagnostic criteria remain limited. This review synthesizes existing scientific literature to provide a concise yet comprehensive overview of the prevalence, risk factors, and clinical diagnostic methods of NSBVAs in pre-presbyopic and presbyopic populations, with an in-depth examination of age-related changes in binocular vision function among older adults. By integrating and comparing analyses of the correlations between age and various binocular vision parameters, this review aims to enhance understanding of age-related binocular vision dysfunction and establish a theoretical foundation for developing targeted diagnostic and therapeutic approaches. Ultimately, these insights could contribute to improved visual performance and life satisfaction in elderly individuals.

BACKGROUND: The effects of acupuncture have varied in different randomized controlled trials (RCTs), and there are many factors that influence treatment effect of acupuncture in different outcomes, with conflicting results. OBJECTIVE: To identify factors and their impact on the treatment effect of acupuncture in different outcomes. METHODS: Acupuncture RCTs were searched from 7 databases including Medline (PubMed), Embase, Cochrane Central Register of Controlled Trials, China National Knowledge Infrastructure, Wanfang Database, VIP Database, and China Biology Medicine disc between January 1st, 2015 and December 31st, 2019. Eligible studies must compare acupuncture to no acupuncture, sham acupuncture, or waiting lists, and report at least 1 patient-important outcome. A multi-level meta-regression was conducted using a 3-level robust mixed model and univariate analyses were performed for all independent variables, even those excluded from the multivariable model due to collinearities. We used thresholds of 0.2 and 0.4 for the difference of standardized mean differences (SMDs), categorising them as small (<0.2), moderate (0.2-0.4), or large (>0.4) effects. RESULTS: The pain construct analysis involved 211 effect estimates from 153 studies and 14 independent variables. High-frequency acupuncture treatment sessions produced larger effects compared to low-frequency sessions [large magnitude, the difference of adjusted SMDs 0.46, 95% confidence interval (CI) 0.07 to 0.84; P=0.02]. The non-pain symptoms construct analysis comprised 323 effect estimates from 231 studies and 15 independent variables. Penetrating acupuncture showed moderately larger effects when compared to non-penetrating acupuncture (0.30, 95% CI 0.06 to 0.53; P=0.01). The function construct analysis included 495 effect estimates from 274 studies and 14 independent variables. Penetrating acupuncture and the flexible acupuncture regimen showed moderately larger effects, compared to non-penetrating acupuncture and fixed regimen, respectively (0.40, 95% CI 0 to 0.80; P=0.05; 0.29, 95% CI 0.06 to 0.53; P=0.01). CONCLUSIONS High-frequency acupuncture sessions appear to be a more effective approach to managing painful symptoms. Penetrating acupuncture demonstrated greater effect in relieving non-painful symptoms. Both penetrating acupuncture type and flexible acupuncture regimen were linked to significant treatment effects in function outcomes. Future studies should consider the factors that are significantly associated with the effects of acupuncture in patient-important outcomes.
Humans ; Randomized Controlled Trials as Topic ; Acupuncture Therapy/methods* ; Pain ; Pain Management ; China

Recent clinical studies have shown that mutation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene in cancer cells may be associated with immunosuppressive tumor microenvironment (TME) and poor response to immune checkpoint blockade (ICB) therapy. Therefore, efficiently restoring PTEN gene expression in cancer cells is critical to improving the responding rate to ICB therapy. Here, we screened an adeno-associated virus (AAV) capsid for efficient PTEN gene delivery into B16F10 tumor cells. We demonstrated that intratumorally injected AAV6-PTEN successfully restored the tumor cell PTEN gene expression and effectively inhibited tumor progression by inducing tumor cell immunogenic cell death (ICD) and increasing immune cell infiltration. Moreover, we developed an anti-PD-1 loaded phospholipid-based phase separation gel (PPSG), which formed an in situ depot and sustainably release anti-PD-1 drugs within 42 days in vivo. In order to effectively inhibit the recurrence of melanoma, we further applied a triple therapy based on AAV6-PTEN, PPSG^@anti-PD-1 and CpG, and showed that this triple therapy strategy enhanced the synergistic antitumor immune effect and also induced robust immune memory, which completely rejected tumor recurrence. We anticipate that this triple therapy could be used as a new tumor combination therapy with stronger immune activation capacity and tumor inhibition efficacy.

Antibiotic-associated diarrhea(AAD)in children is a type of diarrhea that occurs after the use of antibiotics in children,and its pathogenesis is closely related to the intestinal flora.The medication of antibiotics can affect the metabolic function of the intestinal flora and the immune function of the body,and then leads to the occurrence of AAD.In the view of Chinese medicine,AAD in children is mainly involved the spleen,and the etiology of the disease is due to the weakness of the spleen and stomach of the body constitution together with the attack of the pestilential pathogen and the accumulation of drug toxin.The pathogenesis of ADD in children is characterized by spleen deficiency with predominant dampness,deficiency of spleen qi,and insufficiency of spleen yang.Spleen deficiency is the root cause of pediatric AAD,and spleen and intestinal flora have commonality,so the treatment of pediatric AAD can be performed from the perspective of the spleen.The treatment of pediatric ADD from the spleen follows the principle of strengthening and activating the spleen,and the regulation of the spleen for achieving the purpose of treating the disease from the root can be achieved by the methods of strengthening spleen and draining dampness,strengthening spleen and replenishing qi,and strengthening spleen and warming yang separately with the fundamental prescriptions of Shenlin Baizhu Powder,Sijunzi Decoction,and Fuzi Lizhong Pills.

BACKGROUND:The implant osseointegration rate of patients with diabetes is low,and the failure rate is high,which seriously affects the quality of life.It is urgent to improve the implant osseointegration of patients with diabetes by effective means to elevate the success rate.Exploring the effect of berberine on the osteogenic differentiation of bone marrow mesenchymal stem cells under a high-glucose environment and its specific mechanism will provide effective theoretical support for solving the above problems. OBJECTIVE:To explore the effect of natural extract berberine on the osteogenic differentiation of rat bone marrow mesenchymal stem cells under the high-glucose microenvironment. METHODS:Bone marrow mesenchymal stem cells of SD rats were cultured by the whole bone marrow adherence method.CCK-8 assay was used to detect the effects of different concentrations of berberine on the proliferation of bone marrow mesenchymal stem cells under the high-glucose environment and to screen out the optimal berberine concentration.The expressions of Runx2 and Osx were detected by alkaline phosphatase activity,alicarin red staining and PCR to determine the effect of berberine on osteogenic differentiation of bone marrow mesymal stem cells under the high-glucose environment.To further explore the underlying mechanism,we introduced the AMPK-specific inhibitor Dorsomorphin and used a DCFH-DA reactive oxygen species fluorescent probe to examine reactive oxygen species levels.The p-AMPK expression was also determined by western blot assay. RESULTS AND CONCLUSION:(1)10 μmol/L was the optimal concentration of berberine to promote bone marrow mesenchymal stem cell proliferation.(2)Alberberine promoted alkaline phosphatase viability of bone marrow mesenchymal stem cells and mineralized nodule formation in a high-glucose microenvironment.(3)Alberberine promoted the expression of Runx2 and OSx in a high-glucose microenvironment.(4)Alberensine effectively inhibited the reactive oxygen species level of bone marrow mesenchymal stem cells in a high-glucose environment.(5)The effects of berberine on promoting bone marrow mesenchymal stem cell osteogenesis and inhibition of reactive oxygen species were reversed by the AMPK inhibitor.(6)Berberine activated AMPK and promoted p-AMPK expression.(7)The above results indicate that berberine(10 μmol/L)promotes the osteogenic differentiation of bone marrow mesenchymal stem cells in a high-glucose environment by activating AMPK and reducing intracellular reactive oxygen species levels.

BACKGROUND:Proximal humerus internal locking system locking plate is the most commonly used fixation method for proximal humeral fractures,but its failure rate is still high in clinical practice.Reasonable screw placement is an important factor affecting the stability of internal fixation. OBJECTIVE:To investigate the distribution of humeral head screws in the treatment of proximal humeral fractures with proximal humerus internal locking system plate and its influence on internal fixation failure. METHODS:From January 2017 to December 2021,data from patients with proximal humeral fractures undergoing proximal humerus internal locking system plate in Third Affiliated Hospital of Guangzhou University of Chinese Medicine were retrospectively analyzed.A total of 124 patients were enrolled,including 16 males and 108 females,at the age of≥60 years.According to whether there was internal fixation failure after operation,they were divided into normal group(n=101)and internal fixation failure group(n=23).The patient's age,gender,fracture type,the integrity of the medial column,plate height,neck-shaft angle,whether the talus screw was inserted,and the number of humeral head screws,were collected.The humeral head was divided into eight zones according to the postoperative digital radiography anteroposterior film,and the distribution characteristics of the screws in each zone were collected,and the heat map of the screw distribution was drawn. RESULTS AND CONCLUSION:(1)There were no significant differences between the two groups in age,gender,fracture type,the integrity of the medial column,plate height,neck-shaft angle,whether to insert talus screws,and the number of humeral head screws(P>0.05).(2)The heat map showed that the humeral head screws were evenly distributed in the normal group,mainly scattered in zones 4,6,and 7.However,the screw distribution in the internal fixation failure group was not uniform,mainly concentrated in zones 4 and 6.In addition,in the ideal area of talus screws(7/8 zone),there were significantly more screws in the normal group than in the internal fixation failure group.(3)It is indicated that in the treatment of proximal humeral fractures with proximal humerus internal locking system plate,the uniform distribution of humeral head screws is the key to ensuring the internal fixation effect.A reasonable distribution of humeral head screws helps to improve the treatment effect and the success rate of internal fixation.

Objective To explore the role of mitochondrial autophagy in ovarian inflammation associated with poly-cystic ovary syndrome(PCOS)based on the NOD-like receptor thermoprotein domain-related protein 3(NLRP3)pathway.Methods Human ovarian granulosa cell line SVOG was treated with 25 nmol/L dihydrotestosterone(DHT)for 24 h to establish PCOS cell model.SVOG cells were transfected with adenovirus carrying NLRP3(Ad-NLRP3)and negative vector(Ad-EV)or NLRP3 shRNA(sh-NLRP3)and negative control(sh-NC)to overex-press or knockdown NLRP3.Mito-Tracker staining and GFP-LC3 staining were used to evaluate mitochondrial auto-phagy in cells.TUNEL staining,JC-1 staining and Mito-SOX staining were used to analyze the apoptosis,mito-chondrial membrane potential and mitochondrial-derived superoxide production.32 female BALB/c mice were ran-domly divided into three groups:control(Con)group,DHEA group,DHEA+sh-NC group and DHEA+sh-NL-RP3 group,with 8 mice in each group.Except the control group,all other groups treated mice with dehydroepi-androsterone(DHEA)to establish PCOS mouse model.DHEA+sh-NLRP3 group and DHEA+sh-NC group were administrated with sh-NLRP3 or sh-NC encapsulated in lentivirus at a concentration of 1 x 109 TU/ml via tail vein injection.The ultrastructure of mitochondria in ovarian tissue of mice in each group was observed by transmission e-lectron microscope.Results Compared with DHT+sh-NC group,the level of NLRP3 of SVOG cells in DHT+sh-NLRP3 group decreased(P＜0.05).The co-location of GFP-LC3 and mitochondria in SVOG cells in DHT+sh-NLRP3 group was higher than that in DHT+sh-NC group(P＜0.05).Compared with DHT+sh-NC group,the number of TUNEL positive cells and Mito-SOX fluorescence density of SVOG cells in DHT+sh-NLRP3 group de-creased,and the ratio of polymer JC-1 to monomer JC-1 increased(P＜0.05).Compared with Con+Ad-EV group,the level of NLRP3,the number of TUNEL-positive cells and the fluorescence density of mito-SVOG in Con+Ad-NLRP3 group increased(P＜0.05),and the co-location level of GFP-LC3 and mitochondria decreased;the ratio of polymer JC-1 to monomer JC-1 decreased(P＜0.05).Compared with the control group,TUNEL positive cells,relative ROS intensity and percentage of damaged mitochondria in the ovarian tissue of mice in DHEA group increased(P＜0.05).Compared with DHEA+sh-NC group,TUNEL positive cells,relative ROS intensity and percentage of damaged mitochondria in DHEA+SH-NLRP group decreased(P＜0.05).Conclusion Inhibition of mitochondrial autophagy induced by activation of NLRP3 leads to mitochondrial dysfunction and promotes mito-chondrial-related apoptosis in GCs.Knockdown of NLRP3 is beneficial to mitochondrial homeostasis and improves the resistance of GCs to oxidative stress injury,thus promoting the recovery of PCOS.