Objective To elucidate the changes in the gut microbiota and molecular mechanism of huaier in enhancing the efficacy of oxaliplatin (OXA) chemotherapy in gastric cancer (GC). Methods The effects of huaier on the gut microbiota structure and intestinal barrier function in MKN45-bearing mice were analyzed by using 16S rRNA sequencing, RT-qPCR, and IHC. UPLC-MS and network pharmacology, as well as in vivo experimental approaches, were employed to investigate the key bioactive constituents of huaier systematically and elucidate the underlying mechanisms by which huaier exerts therapeutic effects against GC. The expression of the PI3K/AKT/SREBP pathway was detected in cancer cells and tissues via Western blot analysis. The safety profile of huaier combined with OXA was verified through serum biochemistry and HE-stained tissue section analyses. Results Huaier inhibited the PI3K/AKT/SREBP pathway by increasing the abundance of Akkermansia muciniphila, reduced lipid droplet deposition, and ultimately enhanced the sensitivity to OXA in the treatment of GC. Conclusion This study demonstrates the value of huaier in gastric cancer treatment by enhancing chemosensitivity and provides novel insights into its systemic therapeutic effects through molecular mechanisms and gut microbiota modulation.

Objective: To investigate the association between plant-based dietary patterns and gestational weight gain (GWG) among pregnant women with gestational diabetes mellitus (GDM), so as to provide the evidence for guiding the reasonable diet during pregnancy. Methods: GDM pregnant women who participated in the WeBirth project in Hangzhou Obstetrics and Gynecology Hospital were selected. Maternal age and pre-pregnancy body mass index (BMI) were collected. The Chinese version of Pregnancy Physical Activity questionnaire was used to assess the daily activity equivalent. The food frequency questionnaire was used to collect the frequency and amount of food intake in the last month before enrollment. The overall plant-based diet index (PDI), healthy plant-based diet index (HPDI), and unhealthy plant-based diet index (UPDI) were constructed based on food intake and grouped by quartiles. Multiple linear regression models were used to analyze the association between plant-based dietary patterns and GWG. Results: A total of 1 943 pregnant women with GDM, with a median age of 30.91 (interquartile range, 4.92) years. The median BMI of pre-pregnancy was 21.51 (interquartile range, 4.06) kg/m2. The medians of PDI, HPDI and UPDI were 32.42 (interquartile range, 4.60), 32.48 (interquartile range, 4.41) and 32.40 (interquartile range, 5.36), respectively. The median of GWG was 11.30 (interquartile range, 4.52) kg. Multiple linear regression analysis showed that PDI (Q3 group, β=0.674, 95%CI: 0.064-1.285; Q4 group, β=0.702, 95%CI: 0.098-1.306), UPDI (Q3 group, β=1.332, 95%CI: 0.771-1.894; Q4 group, β=1.115, 95%CI: 0.550-1.681) were positively associated with GWG after adjusting for age, pre-pregnancy BMI, daily activity equivalent and daily energy intake. No significant association was found between HPDI and GWG (all P>0.05). Conclusion UPDI was associated with a higher risk of GWG in pregnant women with GDM.

OBJECTIVES: To clarify the role of hippocampal glutamate system in regulating HPA axis in mediating the effect of electroacupuncture (EA) at the heart meridian for improving myocardial injury in rats with acute myocardial ischemia (AMI). METHODS: Male SD rats were randomized into sham-operated group, AMI group, EA group, and L-glutamic acid+EA group (n=9). Rat models of AMI were established by left descending coronary artery ligation, and EA was applied at the "Shenmen-Tongli" segment; the rats in L-glutamic acid+EA group were subjected to microinjection of L-glutamic acid into the bilateral hippocampus prior to AMI modeling and EA treatment. Cardiac functions of the rats were evaluated using echocardiography, and ECG and heart rate variation (HRV) were analyzed using PowerLab and LabChart. Pathological changes in the myocardial tissue was examined using HE staining, and serum levels of myocardial enzymes were detected with ELISA. Myocardial expressions of TH and GAP43 were detected with immunohistochemistry, and colocalization of VGLUT1, VGLUT2 and c-fos were observed using immunofluorescence staining; the expressions of VGLUT1, VGLUT2, NMDAR1 and NMDAR2B were detected using Western blotting. RESULTS: The rat models of AMI showed significantly decreased LVEF and LVFS and increased serum levels of myocardial enzymes in positive correlation with the HPA axis. Numerous TH- and GAP43-positive cells were observed in the hippocampus, where the expressions of NE and E, neurons colabeled with VGLUT1, VGLUT2 and c-fos, and expressions of VGLUT1, VGLUT2, NMDAR1, NMDAR2B and Glu increased significantly. All these changes were significantly improved by interventions with EA as compared with those in AMI and L-Glutamate+EA groups. CONCLUSIONS In rats with AMI, EA at the heart meridian can regulate excessive glutamate release in the hippocampus, thereby inhibiting HPA axis hyperactivity and reducing sympathetic nerve activity to protect the myocardial tissue.
Animals ; Electroacupuncture ; Male ; Rats, Sprague-Dawley ; Hippocampus/metabolism* ; Rats ; Glutamic Acid/metabolism* ; Myocardial Ischemia/physiopathology* ; Hypothalamo-Hypophyseal System/physiopathology* ; Pituitary-Adrenal System/physiopathology* ; Receptors, N-Methyl-D-Aspartate/metabolism*

Cadmium is one of the heavy metals with severe reproductive toxicity,whose half-life lasts 20 to 40 years.Cadmium could induce dysfunction of testis and epididymis for its significant accumulation in human testis,and the amount,motility parameters and morphology of sperm change abnormally.The adverse change could also extend to male offspring and cause the impairment of their reproductive system.There has been no clear mecha-nism of how cadmium induces dysfunction of male reproductive system,and treatment for the adverse influence on male reproductive system by cadmium has not yet been found.Therefore,this problem has been discussed in repro-ductive and environmental field for a long time.A number of previous investigations showed that cadmium could damage male fertility by followed pathways,including interfering with hormone secretion,inducing oxidative stress,activating inflammation and apoptosis,and causing energy metabolism disorder,etc.In order to enlighten new ideas for therapeutic targets of male reproductive function damage induced by cadmium,we systematically reviewed and summarized the findings of previous publications in this paper.

Objective To observe the effects of electroacupuncture(EA)on glutamate(Glu),metabotropic glutamate receptor 2/3(mGluR2/3)and apoptosis related proteins expression in hippocampus in rats with acute myocardial ischemia(AMI);To explore the mechanism of EA against AMI.Methods Totally 50 SD rats were randomly divided into sham-operation group,model group,EA group and inhibitor group,with 10 rats in each group.Except for the sham-operation group,the rats were treated with ligation at the left anterior descending coronary artery to establish AMI model.The rats in the EA group was treated with EA at"Shenmen"and"Tongli",30 minutes each time,once a day for 3 consecutive days.The rats in the inhibitor group were treated with injection of LY341459 via the lateral ventricle 30 min after modeling.HE staining was used to observe myocardial tissue morphology,and ELISA was used to detect Caspase-3 activity in myocardial tissue and Glu content in hippocampal tissue,immunofluorescence staining was used to detect mGluR2/3 expression in hippocampal tissue,TUNEL staining was used to detect apoptosis in hippocampal tissue cells,Western blot was used to detect the expressions of PI3K,Akt,and Caspase-3 protein in hippocampal tissue.Results Compared with the sham-operation group,the myocardial cells of the model group rats showed sparse and swelling with severe infiltration of inflammatory cells;the activity of Caspase-3 in myocardial tissue significantly increased,and the Glu content,positive expression of mGluR2/3,number of apoptotic cells in hippocampal tissue significantly increased(P<0.01),and the expressions of PI3K and Akt proteins in hippocampal tissue were significantly decreased,while the expression of Caspase-3 protein significantly increased(P<0.01).Compared with the model group,myocardial cell edema and inflammatory cell infiltration were reduced in the EA group and inhibitor group,the activity of Caspase-3 in myocardial tissue was significantly decreased,the Glu content,positive expression of mGluR2/3,and number of apoptotic cells in hippocampal tissue were significantly reduced(P<0.01),the expressions of PI3K and Akt proteins in hippocampal tissue significantly increased,while the expression of Caspase-3 protein significantly decreased(P<0.01).Conclusion EA can improve myocardial injury in AMI rats,and its mechanism may be related to activation of PI3K/Akt signaling pathway,inhibition of hippocampal mGluR2/3 overexpression,reduction of Glu accumulation,inhibition of apoptosis of hippocampal neurons and reduction of neurotoxicity.

Objective To construct a culture system of endometrial organoids for infertility induced by non-immune or immune factors,and to compare the immune cytokines between them.Methods The samples were collected from infertility patients undergoing hysteroscopy in Department of Reproductive Medicine Center of The First Affiliated Hospital of Naval Medical University(Second Military Medical University).Endometrial tissues were obtained from patients with infertility caused by non-immune factors(n=3)and patients with repeated implantation failure(RIF)(n=5).The tissues were embedded in matrix glue for 3D culture after washing,digestion,re-suspension and plate attachment.The growth of endometrial organoids of the 2 groups was observed under inverted microscope;the expression of estrogen receptor,keratin and E-cadherin,which were specific endometrial markers,was detected by immunofluorescence staining;and the cytokines of the 2 groups were determined by enzyme-linked immunosorbent assay(ELISA)and the differences of cytokines between the 2 groups were observed.Results During the process of in vitro culture of endometrial organoids,the volume of organoids and the number of cells gradually increased.After 7-10 d of culture,the volume of organoids reached a stable state,and the shape gradually became a perfect circle.At the same time,the number of organoids from the infertility patients caused by non-immune factors was more than that from the infertility patients caused by RIF.Immunofluorescence staining showed the expression of endometrial related marker proteins estrogen receptor,keratin and E-cadherin,indicating the successful construction of endometrial organoids.ELISA results showed that the levels of interferon(IFN)-γ,interleukin(IL)-10,tumor necrosis factor(TNF)-α,IL-4 and TNF-α/IL-4 ratio between the 2 groups were significantly different(P＜0.05 or P＜0.01).There were no significant differences in the levels of TGF-β1 or IL-17,the ratios of IL-17/TGF-β1,IFN-γ/IL-10 or TNF-α/IL-10(all P＞0.05).Conclusion Endometrial organoids with proliferative ability from patients with non-immune infertility and RIF have been successfully cultured in vitro,which provides a new model for the basic research of immune infertility.

Objective To investigate the clinical significance of the expression of antioxidant 1 copper chaperone protein(ATOX1)in hepatocellular carcinoma(HCC)and its relationship with tumor proliferation,migration and invasion.Methods The expression of ATOX1 mRNA in HCC cancer tissue and normal liver tissue was analyzed using the Human Genome Atlas database.Immunohistochemical experiment was used to detect the expression of ATOX1 in 15 cases of HCC cancer tissue and adjacent tissue.Human HCC cell lines Hep3B and HepG2 were divided into the control group(NC),the ATOX1 knockdown group 1(si-ATOX1#1)and the ATOX1 knockdown group 2(si-ATOX1#2).The effects of ATOX1 knockdown on the malignant biological behavior of HCC cells were observed through CCK-8 cell proliferation experiment,scratch experiment and Transwell invasion experiments.A nude mouse xenograft tumor model was constructed to analyze the effect of ATOX1 knockdown on the quality and volume of transplanted tumors.Western blot assay was used to detect the relationship between ATOX1 and JAK2/STAT3 pathway protein expression.Results Bioinformatics analysis showed that expression of ATOX1 mRNA in HCC cancer tissue was higher than that in adjacent normal tissue(P＜0.05).The immunohistochemical staining results showed that the positive rate of ATOX1 protein was higher in HCC cancer tissue than that in adjacent tissue(93.33%vs.13.33%,P＜0.01).In vitro experimental results showed that siRNA knockdown of ATOX1 protein expression in Hep3B and HepG2 cells significantly reduced the proliferation,migration and invasion abilities of cancer cells(P＜0.05).In vivo experiments in mice showed that the volume and weight of subcutaneous xenograft tumors were significantly smaller in the sh-ATOX1 group than those in the sh-con group(P＜0.05).The expression levels of JAK2/STAT3 pathway-related proteins p-JAK2,p-STAT3,CyclinD1 and MMP2 were significantly lower in the subcutaneous transplanted tumor tissue of the sh-ATOX1 group than that of the sh-con group(P＜0.05).Conclusion ATOX1 can promote the proliferation,migration and invasion of HCC through JAK2/STAT3 pathway,which can potentially become a potential tumor marker and therapeutic target.

Objective: To analyze the effects of 12week highintensity fitness exercise on body composition,lipid metabolism and gut microbiota in obese adolescents, so as to provide references for improving the health levels of obese adolescents. Methods: From January to June 2023, 20 obese adolescents from Huaifeng Vocational and Technical School in Huaian City were recruited for the study. Participants were assigned to an exercise group (n=10) and a control group (n=10) for a 12week exercise intervention by random number table method, and both groups had the same diet during the intervention period. The exercise group engaged in three exercises every week, mainly consisting of moderate to highintensity aerobic exercise combined with highintensity intervals. In the first week, there was a 30 minutes of aerobic exercise, followed by 10 minutes of highintensity interval training in the total intervention time each week, and the rest of the time was aerobic exercise with a total intervention time of 60 minutes to maintain; the control group did not receive specific interventions. Body composition was measured using bioelectrical impedance analysis, and lipid levels were determined using an automatic biochemical analyzer. The expression levels of serum inflammatory factors were measured at baseline and after 12 weeks of intervention, and gut microbiota was analyzed using 16S rRNA gene sequencing. Statistical analysis was performed using t test and Chisquare test. Results: After 12 weeks of intervention, the levels of triglycerides (TG), and lowdensity lipoprotein cholesterol (LDL-C) in obese adolescents in the exercise group decreased from (1.7±0.6, 3.5±0.8) mmol/L to (0.9±0.3, 2.6±0.4) mmol/L, while highdensity lipoprotein cholesterol (HDL-C) increased from (1.1±0.2) mmol/L to (1.4±0.2) mmol/L; and serum interleukin-1 receptor antagonist(IL-1Rn) decreased from (8.4±1.6) to (4.5±0.4) ng/mL in the exercise group (t=7.34,2.49,-3.05,2.56, P<0.05). The α-diversity results showed that the Chao index (268.00±22.67) and Ace index (243.98±38.64) in the exercise group were higher than those in the control group (184.52±19.28, 171.43±23.33), and the differences were statistically significant (t=2.48, 2.53, P<0.05). The Shannon index (5.36±1.41) in the exercise group was higher than that in the control group (4.73±1.12), and the Simpson index (0.78±0.10) was lower than that in the control group (0.89±0.10), but the differences were not statistically significant (t=1.83, -2.10, P>0.05). The β-diversity results showed that the intergroup differences in gut microbiota structure between the exercise group and the control group were greater than the intragroup differences, and the differences in gut microbiota structure between the exercise group and the control group were statistically significant (R2=0.083，P<0.05). After intervention, there were significant differences in the relative abundances at the levels of phylum, class, genus, and species in gut microbiota among obese adolescents between the exercise group and the control group (P<0.05). Conclusion The 12week highintensity fitness exercise can alleviate obesity symptoms in obese adolescents through the gut microbiota-lipid metabolism pathway and improve mild chronic inflammatory status.

OBJECTIVE To evaluate the economics of trastuzumab deruxtecan versus the physician-selected chemotherapy （TPC） regimen in the second-line treatment of advanced breast cancer with epidermal growth factor receptor 2 （HER-2） low expression from the perspective of the Chinese healthcare system. METHODS Based on the data of DESTINY-Breast04 clinical trial， the dynamic Markov model was constructed. The time frame of the model simulation was 10 years， and the cycle was 3 weeks. Taking cost， quality-adjusted life year （QALY） and incremental cost-effectiveness ratio （ICER） as the model output indicators， the discount rate of 5% was applied， and 3 times China’s per capita gross domestic product （GDP） in 2023 was taken as the willingness-to-pay （WTP） threshold value. Cost-utility analysis was used to evaluate the economics of the two treatment regiments in the hormone receptor-positive cohort and all patient cohorts， and uncertainty analysis was used to verify the robustness of the basic analysis result. RESULTS The results of the basic analysis showed that compared with the TPC regimen， the ICER value of trastuzumab deruxtecan regimen edu.cn were 1 045 655.76 and 906 404.99 yuan/QALY in the hormone receptor-positive cohort and all patients， respectively， both exceeding the WTP threshold （268 074 yuan/QALY）. The results of single factor sensitivity analysis showed that progression-free survival utility value， the price of trastuzumab deruxtecan and progression disease utility had a significant influence on the model results. The results of probability sensitivity analysis showed that when the WTP threshold was 3 times China’s per capita GDP in 2023， the probability of economic viability of trastuzumab deruxtecan was 0. The results of scenario analysis showed that when the patient assistance program for trastuzumab deruxtecan was considered， the probability of trastuzumab deruxtecan regimen being economical was 0. However， when the price of trastuzumab deruxtecan was reduced by 70%， the probability of its being cost-effective was significantly increased to 82.80%. CONCLUSIONS At a WTP threshold of 3 times China’s per capita GDP in 2023， the trastuzumab deruxtecan regimen is not cost-effective compared to TPC regimen for the second-line treatment of advanced breast cancer with HER-2 low expression. Reducing the price of trastuzumab deruxtecan by region can improve its cost-effectiveness.

Objective:To explore the difference of inflammatory factor imbalance between adolescent and adult patients with depression.Methods:A total of 30 adolescent and 30 adult patients with depression, and 30 adolescent and 30 adult healthy controls were included from January 2022 to August 2023. Interleukin-6 (IL-6), interleukin-17 (IL-17), transforming growth factor-beta1(TGF-β1), interleukin-10(IL-10) and Maresin-1(MaR1) level were detected by enzyme-linked immunosorbent assay. 24-item Hamilton depression scale (HAMD-24) was used to assess the severity of depression in all depressed patients. SPSS 26.0 statistical software was used for t-test, covariance analysis, Spearman analysis and multivariate binary logistic regression, and the predictive value of selected inflammatory factors in depression was evaluated by receiver operating characteristic(ROC) curve. Results:(1)In adolescent group, the levels of IL-6 ((64.000±38.632) pg/mL), IL-17((239.132±49.757) pg/mL), and TGF-β1((737.267±328.447)pg/mL) in patients with depression were higher than those in control group((32.396±16.330)pg/mL, (214.954±42.326)pg/mL, (454.542±297.194)pg/mL, all P<0.05), while the level of MaR1((21 381.301±3 946.011)pg/mL) was significantly lower than that in control group((30 130.138±10 278.999)pg/mL)( P<0.001). The level of IL-17 was positively correlated with the total score of HAMD-24 ( r=0.429) and the course of disease ( r=0.571), the level of IL-10 was negatively correlated with body weight factor score ( r=-0.384), and the levels of TGF-β1 was negatively correlated with anxiety/somatization factor score ( r=-0.449)(all P<0.05) in adolescent patients with depression.MaR1( B=0.000 1, OR=0.999 8, AUC=0.794, P<0.05) was an independent risk factor for adolescents depression.(2)In adult depression group, the levels of IL-6, IL-17, IL-10, TGF-β1 and MaR1 were higher than those in adult control group(all P<0.05). The level of TGF-β1 in adult depression group was negatively correlated with the total score of HAMD-24 ( r=-0.427), the score of anxiety/somatization factor ( r=-0.368), the score of blocking factor ( r=-0.405), and the score of hopelessness factor ( r=-0.398).The level of MaR1 was positively correlated with the age of onset of disease ( r=0.425)(all P<0.05) in adult patients with depression.MaR1( B=0.000 4, OR=1.000 3, AUC=0.874, P<0.001) and IL-6( B=0.040, OR=1.040 7, AUC=0.779, P<0.05) were independent risk factors for adult depression.The AUC of IL-6 combined with MaR1 was 0.938. Conclusion:There are differences in the underlying mechanism of immune imbalance between adolescent and adult patients with depression.MaR1 may be a diagnostic biomarker for depression in adolescents and adults.