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.

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 establish an optimized method for isolation and purification of microglia from aged rat brain tissue, and the phenotype of microglia was detected by flow cytometry. Methods: With young rats(3 months old) as control, the brain tissues of aged rats were immediately processed into single cell suspensions by mechanical dissociation and enzymatic digestion using type IV collagenase. Microglia were isolated on Percoll gradients(30%-37%-70%). The cells were stained with fluorescence-labeled antibodies and the phenotype of microglia was detected by flow cytometry. Results: This study developed a method that enzymatic digestion and mechanical dissociation combined with density gradient centrifugation. More single cells could be obtained by using this method. And the survival rate of cells was more than 90%. The flow cytometric analysis showed that the expression of M1 microglia marker CD86 and MHC Ⅱ increased(P<0.01), and the expression of M2 microglia marker CD200R increased(P<0.01) in aged rats compared with that in young rats. Conclusion The use of type IV collagenase and mechanical digestion combined with density gradient centrifugation is good for isolating and purifying microglia from adult and aged rat brain tissue.

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 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.

The stimulator of interferon genes(STING),an integral adaptor protein in the DNA-sensing pathway,plays a pivotal role in the innate immune response against infections.Additionally,it presents a valuable therapeutic target for infectious diseases and cancer.We observed that fangchinoline(Fan),a bis-benzylisoquinoline alkaloid(BBA),effectively impedes the replication of vesicular stomatitis virus(VSV),encephalomyocarditis virus(EMCV),influenza A virus(H1 N1),and herpes simplex virus-1(HSV-1)in vitro.Fan treatment significantly reduced the viral load,attenuated tissue inflammation,and improved survival in a viral sepsis mouse model.Mechanistically,Fan activates the antiviral response in a STING-dependent manner,leading to increased expression of interferon(1FN)and interferon-stimulated genes(ISGs)for potent antiviral effects in vivo and in vitro.Notably,Fan interacts with STING,preventing its degradation and thereby extending the activation of IFN-based antiviral responses.Collectively,our findings highlight the potential of Fan,which elicits antiviral immunity by suppressing STING degra-dation,as a promising candidate for antiviral therapy.

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 investigate the effects of galectin-3(Gal3)on skin abscess development and activation of mast cells(MC)in mice infected with methicillin-resistant Staphylococcus aureus(MRSA).Methods Wild type mice and Gal3-knockout(Gal3-/-)mice,at 6～8 weeks of age,were divided into four groups:Wild type mice+PBS group,Wild type mice+MRSA group,Gal3-/-mice+PBS group,Gal3-/-mice+MRSA group,were subcutaneously injected with MRSA or the same volume of phosphate buffer saline,with five mice per group.The development and pathological changes of skin abscess were monitored and recorded.The bacterial load in skin tissues was compared,and the expression of associated cytokines,degranulation of MC,and the distribution of MC activation marker 5-hydroxytryptamine(5-HT)were detected.Results The skin of Wild type mice showed progressive abscesses after subcutaneous infection with MRSA,but the Gal3-/-mice showed smaller abscess areas.Compared to the Wild type mice+MRSA group,the Gal3-/-mice+MRSA group showed lower bacterial loading in the skin tissues(P＜0.01)and fewer infiltrating inflammatory cells with histopathological observation.The expression of cytokines,including IL-1β,TNF-α,IL-33,TGF-β,and IL-10,were significantly lower in Gal3-/-mice than Wild type mice(P＜0.05).The toluidine blue staining showed a large number of degranulated MCs in the skin tissues of the wild type mice+MRSA group,whereas only a few degranulated MCs were observed in the Gal3-/-mice+MRSA group.It was further found that the expression of 5-HT in Gal3-/-mice+MRSA group was significantly lower than that in wild-type mice+MRSA group with immunohistochemical staining.Conclusion Gal3 deficiency reduced the activation and degranulation of mouse skin MC after MRSA infection,resulting in changes to inflammatory responses and alleviating the severity of skin tissue abscesses.

The purpose of this study was to investigate the damage mechanism of pathogenic E.coli on mouse brain microvascular endothelial cells(BMEC cells)and mouse alveolar macrophages(MH-S cells),as well as the lung and brain of healthy mice.In this study,BMEC cells and MH-S cells were infected with pathogenic E.coli strains,and cell morphological changes were observed.Plate counting method was used to detect the adhesion and invasion ability of the strains to cells and the number of bacteria in the lungs and brains of mice.RT-qPCR was used to detect the ex-pression of TNF-α,IL-1β and IL-6 genes in cells and mouse organs at different time periods.West-ern blot was used to detect the expression of p-NF-κB,p-JAK2 and p-STAT3 proteins related to inflammation in cells and mouse organs after infection.The results showed that the cell culture medium of the infection group was turbid,the cell vision became dark and blurred,some cells shrank and died,and more fragments were produced.The adhesion rate and invasion rate of BMEC cells at 3 h were significantly lower than those at 6 h(P＜0.050),and the adhesion rate and inva-sion rate of MH-S cells at 3 h were significantly higher than those at 6 h(P＜0.010).Infected mice had a large area of swelling and bleeding in the brain,and the lungs had different degrees of swell-ing and bleeding.The bacterial load in the brain and lung was the highest at 12 h.Compared with the control group,the mRNA expression levels of IL-1β,IL-6 and TNF-α in the infection group were significantly increased at 3 h and 6 h(P＜0.050),and the mRNA expression levels of inflam-matory factors in BMEC cells and MH-S cells were the highest at 6 and 3 h,respectively.The mR-NA expression of inflammatory factors in the brain and lung of infected mice showed a trend of in-creasing first and then decreasing with time,with the highest expression at 12 h after infection.The expression levels of p-NF-κB protein in BMEC cells,MH-S cells,lung and brain tissues of mice in the infection group were significantly higher than those in the control group(P＜0.001),and the expression levels of p-JAK2 protein and p-STAT3 protein were significantly lower than those in the control group(P＜0.050).The above results showed that pathogenic E.coli could adhere and invade BMEC cells and MH-S cells,colonize in lung and brain tissues of mice,promote the expres-sion of NF-κB protein in cells and tissues,inhibit the expression of JAK2 protein and STAT3 pro-tein,and then stimulate cells and tissues to produce inflammatory response.