Purpose: Human epidermal growth factor receptor 2 (HER2)-low breast cancer has the potential to emerge as a distinct subtype. Several studies have compared the differences between HER2-low and HER2-0 breast cancers, but no consensus has been reached.Additionally, a biomarker to predict pathological complete response (pCR) rates in patients with HER2-low breast cancer remains to be identified. Methods: We collected data from 777 patients across three centers, stratifying them into HER2-low and HER2-0 groups. We compared differences in survival and pCR rates between the two groups and investigated potential biomarkers that could reliably predict pCR. Results: The study found that patients with HER2-0 breast cancer had higher pCR rates compared to patients with HER2-low tumors (289 patients [30.1%] vs. 475 patients [18.1%], p < 0.0001). Survival analysis showed no significant advantage for HER2-low tumors over HER2-0 breast cancers. Binary logistic analysis revealed that androgen receptor (AR) expression predicts poorer pCR rates in both the overall patient group and the HER2-0 breast cancer group (overall patients: odds ratio [OR], 0.479; 95% confidence interval [CI], 0.250–0.917; p = 0.026 and HER2-0 patients: OR, 0.267; 95% CI, 0.080–0.892; p = 0.032). In contrast, programmed death ligand 1 (PD-L1) expression was associated with more favorable pCR rates in the overall patient group (OR, 3.199; 95% CI, 1.020–10.037; p = 0.046). Conclusion There is currently insufficient evidence to classify HER2-low breast cancer as a distinct subtype. Our study revealed that AR expression, along with negative PD-L1 expression, contributes to lower pCR rates.

Purpose: Human epidermal growth factor receptor 2 (HER2)-low breast cancer has the potential to emerge as a distinct subtype. Several studies have compared the differences between HER2-low and HER2-0 breast cancers, but no consensus has been reached.Additionally, a biomarker to predict pathological complete response (pCR) rates in patients with HER2-low breast cancer remains to be identified. Methods: We collected data from 777 patients across three centers, stratifying them into HER2-low and HER2-0 groups. We compared differences in survival and pCR rates between the two groups and investigated potential biomarkers that could reliably predict pCR. Results: The study found that patients with HER2-0 breast cancer had higher pCR rates compared to patients with HER2-low tumors (289 patients [30.1%] vs. 475 patients [18.1%], p < 0.0001). Survival analysis showed no significant advantage for HER2-low tumors over HER2-0 breast cancers. Binary logistic analysis revealed that androgen receptor (AR) expression predicts poorer pCR rates in both the overall patient group and the HER2-0 breast cancer group (overall patients: odds ratio [OR], 0.479; 95% confidence interval [CI], 0.250–0.917; p = 0.026 and HER2-0 patients: OR, 0.267; 95% CI, 0.080–0.892; p = 0.032). In contrast, programmed death ligand 1 (PD-L1) expression was associated with more favorable pCR rates in the overall patient group (OR, 3.199; 95% CI, 1.020–10.037; p = 0.046). Conclusion There is currently insufficient evidence to classify HER2-low breast cancer as a distinct subtype. Our study revealed that AR expression, along with negative PD-L1 expression, contributes to lower pCR rates.

Purpose: Human epidermal growth factor receptor 2 (HER2)-low breast cancer has the potential to emerge as a distinct subtype. Several studies have compared the differences between HER2-low and HER2-0 breast cancers, but no consensus has been reached.Additionally, a biomarker to predict pathological complete response (pCR) rates in patients with HER2-low breast cancer remains to be identified. Methods: We collected data from 777 patients across three centers, stratifying them into HER2-low and HER2-0 groups. We compared differences in survival and pCR rates between the two groups and investigated potential biomarkers that could reliably predict pCR. Results: The study found that patients with HER2-0 breast cancer had higher pCR rates compared to patients with HER2-low tumors (289 patients [30.1%] vs. 475 patients [18.1%], p < 0.0001). Survival analysis showed no significant advantage for HER2-low tumors over HER2-0 breast cancers. Binary logistic analysis revealed that androgen receptor (AR) expression predicts poorer pCR rates in both the overall patient group and the HER2-0 breast cancer group (overall patients: odds ratio [OR], 0.479; 95% confidence interval [CI], 0.250–0.917; p = 0.026 and HER2-0 patients: OR, 0.267; 95% CI, 0.080–0.892; p = 0.032). In contrast, programmed death ligand 1 (PD-L1) expression was associated with more favorable pCR rates in the overall patient group (OR, 3.199; 95% CI, 1.020–10.037; p = 0.046). Conclusion There is currently insufficient evidence to classify HER2-low breast cancer as a distinct subtype. Our study revealed that AR expression, along with negative PD-L1 expression, contributes to lower pCR rates.

ObjectiveThis study explores the methods of lung tissue extraction and fixation required for pathological studies of fetal rats, based on the unique physiological structure of fetal rat lung tissue and existing lung tissue fixation techniques for adult rats. MethodsSix pregnant adult SD rats at 20.5 days of gestation were subjected to cesarean section to obtain fetal rats. Four healthy fetal rats with similar body weight, vital signs, and respiratory status were selected from each pregnant rat, and they were randomly divided into the following groups using a random number table: direct lung infiltration group, lung infiltration group after intratracheal infusion, whole-body infiltration group of fetal rats, and whole-body infiltration group after intratracheal infusion of fetal rats. To systematically compare and analyze the anatomical morphology under different fixation methods, lung tissues from four groups of fetal rats were harvested, perfused, and fixed, and the gross morphology of lung tissues in each group was observed. Paraffin sections were prepared and stained with Hematoxylin-Eosin (H&E). The histological morphology of the whole lung, alveoli, and bronchi was further examined under optical microscopy. ResultsIn the direct lung infiltration group, the hilar structures were unclear, lung lobation was indistinct, the shape was irregular, lung cavities were small, and alveoli and bronchi were shrunken. In the lung infiltration group after intratracheal infusion, the hilar structures were clear, lobation was pronounced, the shape was regular, lung cavities were large, and alveoli and bronchi were full. Both the whole-body infiltration group and whole-body infiltration group after intratracheal infusion of fetal rats exhibited visible lungs, hearts, skins, and other organs. The lung tissues of both groups showed obvious lobulation, irregular shape, and damage at the margins of lung lobes. In the whole-body infiltration group, the thoracic cavities of the fetus were flattened, lung cavities were small, and alveoli and bronchi were shrunken. In the whole-body infiltration group after intratracheal infusion of fetal rats, the fetal thoracic cavities were full, lung cavities were large, and alveoli and bronchi were relatively full. ConclusionThe lung infiltration after intratracheal infusion method for fetal rat lung tissue fixation outperforms direct lung infiltration, whole-body infiltration of fetal rats, and whole-body infiltration after intratracheal infusion of fetal rats in terms of preservation of the lung tissue's original morphology, paraffin sectioning, staining, and pathological observation and analysis. The embedding, sectioning, and staining processes are also simple and save consumables. Therefore, intratracheal infusion followed by lung infiltration method is recommended for fixation in histopathological observation of fetal rat lung tissue.

Objective To observe the expression of adhesion molecule CD226 on the small intestinal group 3 innate lymphoid cells (ILC3) in mice. Methods The bioinformatics was used to analyze the expression of CD226 on murine ILCs. Small intestinal mucosal lamina propria lymphocytes (LPL) were isolated from wild-type C57BL/6J mice, and the expression of CD226 on ILC1 and ILC3 was detected by flow cytometry. A mouse model of dextran sulfate sodium (DSS)-induced colitis was constructed to observe the changes in the expression of CD226 on ILC3. Results Both ILC1 and ILC3 in the mice small intestine expressed CD226 molecules; the proportion of ILC3 was reduced, while the expression level of CD226 on ILC3 was increased in the colitis model. Conclusion CD226 is expressed on the small intestines of mice, and although the proportion of ILC3 decreases in the DSS-induced colitis, the expression of CD226 on ILC3 increases.
Animals ; Mice ; Colitis/chemically induced* ; Immunity, Innate ; Intestine, Small ; Lymphocytes ; Mice, Inbred C57BL

ObjectiveTo investigate the possible mechanism of electroacupuncture at "Neiguan" （PC6） and "Zhongwan" （RN12） in the treatment of functional dyspepsia （FD）. MethodsSPF male SD rats were randomly divided into model group， Neiguan （PC6） group， Zhongwan （RN12） group， and Neiguan-Zhongwan （PC6-RN12） group， with 8 rats in each group. Except for the normal group， the other groups of rats were cogavaged with 0.1% sucrose iodoacetamide solution combined with small platform standing training to establish FD rat models. After successful modeling， the rats in the normal group and model group were tied up for 30 min/d for 7 days； the Neiguan group， Zhongwan group， and Neiguan-Zhongwan group were treated with electroacupuncture intervention at "Neiguan" （PC6）， "Zhongwan" （RN12）， and "Neiguan" - "Zhongwan" （PC6-RN12） acupoints， respectively， using continuous wave， frequency of 2 Hz， current intensity of 1 mA， 30 min/d， and continuous intervention for 7 days. The general condition of rats in each group was observed. After treatment， the body weight and food intake of rats were measured， and the gastric emptying rate was calculated； HE staining was performed on the gastric antrum tissue of rats to observe the histopathologic changes； the expressions of calcitonin gene-related peptide （CGRP） and Ghrelin protein in gastric antrum were detected by Western Blot； the metabolites in gastric antrum tissues were detected by liquid chromatography-mass spectrometry （LC-MS）， and differential metabolites were screened by correlation analysis， then Metabo Analyst 5.0 and KEGG databases were used for metabolic pathway analysis. ResultsUnder light microscope， the gastric antrum structure was complete and the glands were abundant. No obvious inflammation and edema were found in gastric mucosa. Compared with the normal group， the body weight， food intake， and gastric emptie rate of rats in model group decreased， the expression of Ghrelin protein decreased and the expression of CGRP protein increased in gastric antrum tissue （P＜0.01）. Compared with model group， the body weight， food intake， and gastric emptance rate of rats in Neiguan group， Zhongwan group and Neiguan-Zhongwan group all increased， CGRP protein expression decreased in Neiguan group， and Ghrelin protein expression increased and CGRP protein expression decreased in Zhongwan group and Neiguan-Zhongwan group （P＜0.01 or P＜0.05）. Compared with Neiguan-Zhongwan group， Ghrelin protein expression decreased and CGRP protein expression increased in Neiguan group and Zhongwan group （P＜0.05 or P＜0.01）. Metabolomics results showed that compared with normal group， the content of metabolites adenosine diphosphate ribose， adenosine monophosphate and adenosine diphosphate in gastric antrum tissue of model group decreased； compared with model group， the contents of adenosine diphosphate， adenosine diphosphate and citicoline in Neiguan group increased， the contents of nicotine adenine dinucleotide， cytidine diphosphate ethanolamine and citicoline in Zhongwan group increased， and the contents of adenosine diphosphate， cytidine diphosphate and citicoline in Neiguan-Zhongwan group increased （P＜0.05 or P＜0.01）. Compared with model group， the main metabolic pathways of different metabolites in PC6-RN12 group were glycerophospholipid metabolism， taurine and hypotaurine metabolism， purine metabolism， starch and sucrose metabolism. ConclusionElectroacupuncture at “Neiguan” and “Zhongwan” acupoints can effectively regulate gastrointestinal motility and improve FD symptoms in FD rats， and the effect is better than that of "Neiguan" or "Zhongwan" acupoints alone. The mechanism may be related to the influence of related metabolites on energy metabolism， glucose metabolism and nucleotide metabolism， thereby regulating gastrointestinal motility hormones.

Objective:To observe the effects of acupuncture and moxibustion at Feishu(BL13)on inflammatory responses and intestinal short-chain fatty acids(SCFAs)in rats with asthma. Methods:Fifty-six Sprague-Dawley rats were randomly divided into a normal group(16 rats)and a modeling group(40 rats).Rats in the modeling group were subjected to establishing asthma models using ovalbumin(OVA).Model evaluation was conducted using 4 rats from each group.The remaining rats that successfully developed asthma were then randomly divided into a model group,an acupuncture group,and a moxibustion group,with 12 rats in each group.Rats in the acupuncture group received acupuncture treatments,and those in the moxibustion group received moxibustion treatments,both at Feishu(BL13)for 30 min.Following the treatments,the rats were exposed to atomization excitation with a 1%OVA solution for 20 min daily for 14 consecutive days.At the end of the experiment,inflammatory markers in the rats'peripheral blood were analyzed using a biochemical method.In addition,inflammatory cells in the bronchoalveolar lavage fluid(BALF)were counted using Wright-Giemsa staining.The lung tissue of rats was examined under a light microscope after staining with hematoxylin-eosin to observe morphological or pathological changes.Furthermore,real-time fluorescence quantitative polymerase chain reaction was utilized to measure the mRNA expression of inflammatory factors in the lung tissue.Lastly,the concentration of SCFAs in the rat's feces was determined using gas chromatography-hydrogen flame ionization. Results:The levels of eosinophils(Eos),neutrophils(Neu),and lymphocytes(Lym)in the peripheral blood,as well as Eos and Neu in the BALF,and the expression of interleukin(IL)-4,IL-5,IL-13,IL-33,and thymic stromal lymphopoietin(TSLP)mRNAs in the lung tissue were all found to be significantly increased(P<0.05 or P<0.01);the lung tissue structure displayed severe injuries;the levels of acetic acid,propionic acid,isobutyric acid,butyric acid,and valeric acid in the feces decreased significantly in the model group(P<0.05 or P<0.01).Compared with the model group,the peripheral blood levels of Eos,Neu,and Lym,as well as Eos in the BALF,and the mRNA expression levels of IL-4 and IL-5 in the lung tissue decreased significantly in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).This reduction was accompanied by alleviated pathological damage in the lung tissue.Additionally,there were significant increases in the levels of acetic acid,propionic acid,isobutyric acid,and butyric acid in the feces in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).In the acupuncture group,the expression levels of Lym in the BALF and IL-13 mRNA in the lung tissue decreased significantly(P<0.05 or P<0.01).In the moxibustion group,the mRNA expression levels of IL-33 and TSLP in the lung tissue also reduced significantly(P<0.05 or P<0.01).Furthermore,the level of valeric acid in the feces increased notably in the moxibustion group(P<0.01).Compared with the acupuncture group,it was found that the mRNA levels of IL-5 and IL-13 in the lung tissue,as well as the acetic acid level in the feces,were significantly higher in the moxibustion group(P<0.05 or P<0.01). Conclusion:Both acupuncture and moxibustion were effective in reducing abnormal inflammation and regulating intestinal SCFAs in asthma model rats.Acupuncture demonstrated superiority in inhibiting pro-inflammatory factors,particularly IL-5 and IL-13,while moxibustion exhibited better regulation on intestinal metabolites SCFAs,especially acetic acid.

Background and purpose:Esophageal carcinoma(ESCA)is one of the malignant tumors with high mortality rate,and the underlying mechanism of its development is largely unknown.CDC20 plays an important role in tumorigenesis,and its dysregulated expression is closely related to tumor occurrence and development.The expression of CDC20 is increased in a variety of tumors,and knocking down CDC20 can inhibit tumor cell proliferation.NLRP3 is the main component of the inflammasome,and inflammasome is also closely related to tumor occurrence and development.Here,our study aimed to investigate whether CDC20 promotes the proliferation of ESCA cells through NLRP3 and its regulatory mechanism.Methods:The expression levels of CDC20 and NLRP3 genes in ESCA patients were analyzed using The Cancer Genome Atlas(TCGA)detabase and GTEx public database.We collected clinical and pathological data and tissues from 80 ESCA patients at the First Affiliated Hospital of Xinxiang Medical College,and detected the protein expression of NLRP3 in ESCA patients through immunohistochemistry staining.This study was approved by the Ethics Committee of the First Affiliated Hospital of Xinxiang Medical College(Number:EC-021-137).We studied the effects of knocking down CDC20 and NLRP3 gene on the proliferation ability of esophageal squamous cell carcinoma cells EC9706 and KYSE150 using short hairpin RNA(shRNA)technology.Co-immunoprecipitation(Co-IP),proteasome inhibitors and ubiquitination experiments were used to detect whether CDC20 interacts with NLRP3,and to elucidate whether CDC20 regulates NLRP3 expression through the ubiquitination pathway.This study was approved by the Ethics Committee of the First Affiliated Hospital of Xinxiang Medical College(Number:EC-021-137).Results:The TCGA database analysis showed that the expression levels of CDC20 and NLRP3 mRNA were significantly higher in the cancer tissues of ESCA patients than in the adjacent tissues.The immunohistochemistry results further showed that compared with adjacent tissues,the protein expression levels of CDC20 and NLRP3 were increased in ESCA tissues.Knocking down CDC20 and NLRP3 genes inhibited the proliferation of ESCA cells.Co-IP,proteasome inhibitors and ubiquitination experiments confirmed that CDC20 interacted with NLRP3 through its leucine-rich repeat(LRR),and CDC20 stabilized its expression by promoting NLRP3 ubiquitination.Conclusion:CDC20 and NLRP3 are upregulated in ESCA tissues,and CDC20 stabilizes their expression through ubiquitination of NLRP3,promoting ESCA cell proliferation.This suggests that CDC20 and NLRP3 may be potential diagnostic targets for ESCA.

ObjectiveTo explore the possible mechanism of moxibustion at "Feishu （BL13）" and "Tianshu （ST25）" in treatment of bronchial asthma. MethodsA total of 48 SD rats were randomly divided into normal group （n=12） and modeling group （n=36）. The bronchial asthma rat model was established by sensitization with ovalbumin （OVA） injection and aerosol provocation. Thirty-two successfully modeled rats were further randomly divided into four groups including model group， Feishu group， Tianshu group， and Feishu-Tianshu group， with 8 rats in each group. Rats in the normal group and the model group were tied and fixed without intervention， while those in the Feishu group received moxibustion at bilateral of "Feishu （BL13）" for 30 minutes； rats in the Tianshu group received moxibustion at bilateral "Tianshu （ST25）" for 30 minutes， and those in the Feishu-Tianshu group received moxibustion at both "Feishu （BL13）" and "Tianshu （ST25）" bilaterally for 15 minutes each. One hour after the intervention， 1% OVA solution was aerosolized for 20 minutes in all groups except the normal group， which was given the same volume of 0.9% sodium chloride solution instead of OVA solution for aerosol stimulation. The above interventions were performed once daily for 14 days. Behavioral observations were performed after modeling and during the interventions. The samples were collected 24h after the last intervention. HE and Masson staining were used to observe pathological morphological changes of lung tissues， and the percentage of collagen fiber deposition area was counted. The levels of leukocyte differentiation antigen11b （CD11b）， leukocyte differentiation antigen 40 （CD40）， leukocyte differentiation antigen 86 （CD86）， and programmed death ligand 2 （PD-L2） in serum， as well as the expression of interleukin-8 （IL-8）， interleukin-11 （IL-11）， interleukin-27 （IL-27） in lung tissue， were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression of matrix metalloproteinase-9 （MMP-9）， matrix metalloproteinase inhibitor 1 （TIMP-1） and transforming growth factor β （TGF-β） proteins in lung tissue. The content of six short-chain fatty acids （SCFAs） including acetic acid， propionic acid， isobutyric acid， butyric acid， valeric acid， and hexanoic acid in feces was detected by gas chromatography-mass spectrometry. ResultsCompared to the normal group， the rats in the model group gradually showed mental depression or restlessness， dull hair， slow activity， reduced food intake， unformed stool， accompanied by symptoms of shortness of breath and wheezing. The pathological results showed severe abnormalities in lung tissue structure in the model group， including extensive infiltration of inflammatory cells around the bronchi， thickening of the airway smooth muscle layer， and substantial deposition of collagen fibers. Significant increases were observed in the levels of serum CD11b， CD40， CD86， and PD-L2， levels of IL-8， IL-11， and IL-27 in the lung tissue， as well as protein expression levels of MMP-9， TGF-β， and TIMP-1 in lung tissue， while the fecal levels of acetic acid， propionic acid， isobutyric acid， butyric acid， valeric acid， and n-caproic acid significantly decreased （P＜0.05 or P＜0.01）. Compared to those in the model group， the spirit， hair， activity， drinking and eating condition， shortness of breath， and wheezing symptoms of rats in the Feishu group， Tianshu group， and Feishu-Tianshu group were improved； the stool was basically formed， and the pathological morphology of lung tissue were improved； the levels of serum CD11b， CD40， CD86 and PD-L2， the levels of IL-8 and IL-27 in the lung tissue， the percentage of collagen fiber deposition area， and the TGF-β protein expression notably decreased； content of IL-11 and MMP-9 in the lung tissue and protein expression of T1MP-1 in Feishu group and Feishu-tianshu group significantly decreased； content of six SCFAs in the Feishu-Tianshu group increased （P＜0.05 or P＜0.01）. Compared to those in the Feishu group， the percentage of collagen fiber deposition area and TIMP-1 protein expression in lung tissue in the Feishu-Tianshu group significantly decreased， while the fecal levels of acetic acid and butyric acid notably increased （P＜0.05 or P＜0.01）. Compared to those in the Tianshu group， the serum level of CD40 in the Feishu-Tianshu group was significantly reduced， and the percentage of collagen fiber deposition area， the content of IL-11， and the protein expressions of MMP-9， TGF-β and TIMP-1 in the lung tissue notably decreased， while the fecal levels of acetic acid， propionic acid， and butyric acid significantly increased （P＜0.05 or P＜0.01）. ConclusionMoxibustion at "Feishu （BL13）" and "Tianshu （ST25）" exhibits a favorable therapeutic effect on airway remodeling in bronchial asthma rats， and the combined application of "Feishu （BL13）" and "Tianshu （ST25）" acupoints demonstrates a synergistic effect. The mechanism may be related to the regulation of intestinal SCFAs content， influencing the differentiation of immune cells， and reducing airway inflammation.

OBJECTIVE To evaluate the cost-effectiveness of clopidogrel versus aspirin monotherapy regimens for secondary prevention of ischemic stroke and to provide economic evidence and reference for clinical medication and decision-making. METHODS Based on the CAPRIE trial， a Markov model was constructed； the probabilities of risk events， health utility values， and costs of risk event management were obtained from relevant literature. The cycle length was 6 months， and the time horizon was 10 years. A discount rate of 5% per year was applied. The primary outcomes were total costs， quality-adjusted life-years （QALYs）， and incremental cost-effectiveness ratio （ICER）. Cost-utility analysis was performed for above 2 regimens by using TreeAge Pro software. The one-way sensitivity analysis， probabilistic sensitivity analysis and scenario analysis were conducted to validate the robustness of the analyses. RESULTS Compared with the aspirin regimen （325 mg/d of CAPRIE trial dose）， the ICER values of clopidogrel regimen for secondary stroke prevention for 10 years， 20 years and 30 years were 4 284.06， 4 201.20 and 3 986.78 yuan/QALY， respectively， which were E-mail：liuxiaoyanrj@sjtu.edu.cn all less than the willing-to-pay （WTP） threshold of one time 。 China’s per capita gross domestic product （GDP） in 2021. E-mail：scilwsjtu-wb@yahoo.com Compared with the aspirin regimen （clinically recommended dose in China， 100 mg/d）， the ICER values of clopidogrel regimen for stroke secondary prevention for 10 years， 20 years and 30 years were 58 238.27， 42 164.72 and 36 164.77 yuan/QALY， respectively， which were all less than WTP threshold. When comparing with aspirin regimen of 325 mg/d， results of one-way sensitivity analysis showed that the cost of clopidogrel and aspirin， probability of the first recurrence of ischemic stroke were sensitive factors of model. Results of probabilistic sensitivity analysis showed that when WTP was set at one time GDP per capita in China in 2021， clopidogrel had a probability of being cost- effective of about 66.5%. Results of scenario analysis showed that neither changing the time horizon to 10， 20 or 30 years nor using different doses of aspirin （50， 100， 150， 200 or 250 mg/d） would not alter any conclusions. CONCLUSIONS Compared with aspirin monotherapy， clopidogrel monotherapy is more cost-effective for secondary prevention of ischemic stroke.