ObjectiveTo explore the potential mechanism of Xiaoqinglong Decoction （小青龙汤， XD） in the treatment of allergic rhinitis. MethodsForty-five rats were randomly assigned to a control group， a model group， a loratadine group， low-， medium- and high-dose XD groups， and low-， medium- and high-dose Mahuang Decoction and Cang'erzi Powder （麻黄汤合苍耳子散， MDCP） groups. Except for the control group， rats were administered with ovalbumin （OVA） and aluminum hydroxide via intraperitoneal injection for 14 days to establish an allergic rhinitis model. After the 14th-day injection， nasal stimulation was continued with 20 μl of 10% OVA solution to maintain the model. Rats in the control group and the model group received 10 ml/（kg·d） of saline， whereas those in the loratadine group were administered with 0.9 mg/（kg·d） of loratadine. The low-， medium- and high-dose XD groups were administered XD at the dose of 2.7， 5.4， and 10.8 g/（kg·d）， respectively. The low-， medium- and high-dose MDCP groups were administered MDCP at the dose of 2.43， 4.86， and 9.72 g/（kg·d）， respectively. All treatments were administered by gavage once daily for 7 consecutive days. One hour after the final gavage， nasal symptom scores were recorded for all group of rats. The next day， serum levels of immunoglobulin E （IgE）， interleukin-4 （IL-4）， and interleukin-13 （IL-13） were measured. HE staining was used to observe the pathological morphology of the nasal mucosal tissue. Quantitative reverse transcription PCR （RT-qPCR） and Western Blot were performed to assess mRNA and protein expression of thymic stromal lymphopoietin （TSLP） and OX40 ligand （OX40L） in the nasal mucosa. ResultsCompared to the control group， total nasal symptom score in the model group significantly increased （P＜0.01）. HE staining revealed disrupted and adhered cilia， thickened basement membranes， and extensive inflammatory cell infiltration in the nasal mucosa. Serum levels of total IgE， IL-4， and IL-13， as well as TSLP and OX40L mRNA and protein expression in the nasal mucosa， were significantly elevated in the model group （P＜0.05 or P＜0.01）. Compared to the model group， the total nasal symptom scores in all drug intervention groups were significantly reduced； the serum total IgE levels in the loratadine group， the low- and medium-dose XD groups， and the low- and high-dose MDCP groups were significantly reduced； and the serum levels of IL-4 and IL-13 in the high-dose XD group and the high-dose MDCP group decreased （P＜0.05 or P＜0.01）. Nasal mucosal structure was improved. Except for the low-dose MDCP group， all other intervention groups showed a significant reduction in TSLP and OX40L mRNA expression in the nasal mucosa （P＜0.01）. All doses of XD and the medium- and high-dose MDCP groups significantly decreased the protein levels of TSLP and OX40L （P＜0.05）. The medium-dose XD group exhibited more improvement of nasal symptom scores and greater suppression of expression of TSLP and OX40L mRNA， and TSLP protein levels compared to the loratadine group （P＜0.05）. ConclusionXD may protect nasal mucosa of rats and alleviate allergic rhinitis by suppressing the TSLP/OX40L pathway， thereby attenuating Th2-mediated immune responses.

ObjectiveTo explore the potential mechanism of Xiaoqinglong Decoction （小青龙汤， XD） in the treatment of allergic rhinitis. MethodsForty-five rats were randomly assigned to a control group， a model group， a loratadine group， low-， medium- and high-dose XD groups， and low-， medium- and high-dose Mahuang Decoction and Cang'erzi Powder （麻黄汤合苍耳子散， MDCP） groups. Except for the control group， rats were administered with ovalbumin （OVA） and aluminum hydroxide via intraperitoneal injection for 14 days to establish an allergic rhinitis model. After the 14th-day injection， nasal stimulation was continued with 20 μl of 10% OVA solution to maintain the model. Rats in the control group and the model group received 10 ml/（kg·d） of saline， whereas those in the loratadine group were administered with 0.9 mg/（kg·d） of loratadine. The low-， medium- and high-dose XD groups were administered XD at the dose of 2.7， 5.4， and 10.8 g/（kg·d）， respectively. The low-， medium- and high-dose MDCP groups were administered MDCP at the dose of 2.43， 4.86， and 9.72 g/（kg·d）， respectively. All treatments were administered by gavage once daily for 7 consecutive days. One hour after the final gavage， nasal symptom scores were recorded for all group of rats. The next day， serum levels of immunoglobulin E （IgE）， interleukin-4 （IL-4）， and interleukin-13 （IL-13） were measured. HE staining was used to observe the pathological morphology of the nasal mucosal tissue. Quantitative reverse transcription PCR （RT-qPCR） and Western Blot were performed to assess mRNA and protein expression of thymic stromal lymphopoietin （TSLP） and OX40 ligand （OX40L） in the nasal mucosa. ResultsCompared to the control group， total nasal symptom score in the model group significantly increased （P＜0.01）. HE staining revealed disrupted and adhered cilia， thickened basement membranes， and extensive inflammatory cell infiltration in the nasal mucosa. Serum levels of total IgE， IL-4， and IL-13， as well as TSLP and OX40L mRNA and protein expression in the nasal mucosa， were significantly elevated in the model group （P＜0.05 or P＜0.01）. Compared to the model group， the total nasal symptom scores in all drug intervention groups were significantly reduced； the serum total IgE levels in the loratadine group， the low- and medium-dose XD groups， and the low- and high-dose MDCP groups were significantly reduced； and the serum levels of IL-4 and IL-13 in the high-dose XD group and the high-dose MDCP group decreased （P＜0.05 or P＜0.01）. Nasal mucosal structure was improved. Except for the low-dose MDCP group， all other intervention groups showed a significant reduction in TSLP and OX40L mRNA expression in the nasal mucosa （P＜0.01）. All doses of XD and the medium- and high-dose MDCP groups significantly decreased the protein levels of TSLP and OX40L （P＜0.05）. The medium-dose XD group exhibited more improvement of nasal symptom scores and greater suppression of expression of TSLP and OX40L mRNA， and TSLP protein levels compared to the loratadine group （P＜0.05）. ConclusionXD may protect nasal mucosa of rats and alleviate allergic rhinitis by suppressing the TSLP/OX40L pathway， thereby attenuating Th2-mediated immune responses.

Background/Aims: Lenvatinib resistance remains a critical barrier in advanced hepatocellular carcinoma (HCC) therapy. However, the underlying mechanisms and strategies for reversing resistance remain incompletely understood. Methods: Integrated transcriptomics of lenvatinib-resistant patient tumors and an acquired-resistance murine model identified a novel macrophage subpopulation. Functional validation employed CRISPR-SAM screening, conditioned medium (CM) assays, subcutaneous/orthotopic xenografts, patient-derived organoids (PDOs), and patient-derived xenografts (PDXs). Mechanistic studies included ChIP-qPCR, co-immunoprecipitation, and pharmacologic targeting. Clinical relevance was assessed in a retrospective cohort. Results: Resistant HCC exhibited significant enrichment of a COLEC12high TAM subset , which correlated with poor survival and treatment response. These TAMs secreted neuregulin-1 (NRG1) , activating HER2/HER3-AKT signaling in tumor cells to drive cancer stemness and lenvatinib resistance. Mechanistically, in TAMs COLEC12 sequestered STAT1 in the cytoplasm, preventing its phosphorylation, and thereby derepressing STAT3-mediated NRG1 transcription. Depletion of NRG1 reversed the stemness phenotypes and resensitized tumors to lenvatinib both in vitro and in vivo. Clinically, high NRG1 expression predicted an inferior lenvatinib response and shorter survival. Crucially, the bispecific anti-HER2/HER3 antibody zenocutuzumab restored lenvatinib efficacy in PDOs, PDXs, and murine models. Conclusions Our work establishes the COLEC12high TAM/NRG1 axis as a master regulator of therapeutic resistance and identifies NRG1 as a predictive biomarker, providing a clinically actionable strategy to overcome lenvatinib resistance in HCC.

ObjectiveTo observe the clinical efficacy of electroacupuncture in treatment of simple obesity patients and its effects on the metabolism-inflammation network. MethodsSeventy-two patients with simple obesity were randomly divided into a electroacupuncture group and a sham electroacupuncture group， with 36 patients in each group. In the electroacupuncture group， conventional acupuncture was applied at Zhongwan （CV 12）， bilateral Tianshu （ST 25）， bilateral Daheng （SP15）， Qihai （CV 6）， Guanyuan （CV 4）， bilateral Zusanli （ST 36） and Fenglong （ST 40）. The sham electroacupuncture group used placebo needle at same acupoints as the electroacupuncture group. Both groups were given sparse-dense waves with a frequency of 2/10 Hz， 30 minutes per session， 3 times a week， for a total of 8 weeks of treatment. The changes of human body parameters including body weight， body mass index （BMI）， waist circumference， and body fat percentage （BFP）， metabolic factors including serum fasting blood glucose （FBG）， triglyceride （TG）， total cholesterol （TC）， high density lipoprotein （HDL）， low density lipoprotein （LDL）， leptin， and ghrelin， and inflammatory factors including interleukin-6 （IL-6） and tumor necrosis factor α （TNF-α） were compared before and after treatment. Correlations between changes in leptin， ghrelin and inflammatory factors were further analyzed. ResultsWaist circumference was significantly reduced after treatment in both groups compared with baseline （P＜0.05）， and the post-treatment waist circumference was lower in the electroacupuncture group than in the sham electroacupuncture group （P＜0.05）； body weight， BMI and BFP decreased significantly after treatment compared to pretreatment levels in the electroacupuncture group （P＜0.05）. Serum leptin， IL-6 and TNF-α declined significantly after treatment in both groups versus baseline （P＜0.05）； the electroacupuncture group had lower post-treatment levels of leptin， IL-6 and TNF-α as well as larger pre-post changes than the sham group （P＜0.05）； ghrelin increased markedly after treatment in both groups （P＜0.05）， with higher post-treatment ghrelin and greater pre-post variation observed in the electroacupuncture group （P＜0.05）. In the electroacupuncture group， changes in ghrelin were negatively correlated with IL-6 and positively correlated with TNF-α （P＜0.01）； in the sham electroacupuncture group， ghrelin changes were negatively correlated with both IL-6 and TNF-α， whereas leptin changes were positively correlated with the two inflammatory markers （P＜0.01）. ConclusionElectroacupuncture can effectively improve clinical manifestations， as well as regulate serum metabolic and pro-inflammatory factors in patients with simple obesity， and discrepant correlation patterns of metabolism-inflammation are observed between the two groups.

Objective:To investigate the epidemic characteristics and trends of major natural focus diseases and vector-borne infectious diseases in Yantai City, and provide a basis for formulating reasonable prevention and control strategies.Methods:The descriptive analysis was conducted on the epidemic data of natural focal diseases and vector-borne infectious diseases in Yantai City collected from the Infectious Disease Surveillance and Reporting Management System, a subsystem of the China Disease Control and Prevention Information System, covering the period from January 1, 2019, to December 31, 2023.Results:From 2019 to 2023, a total of 2 746 cases of 12 kinds of natural focus diseases, vector-borne infectious diseases and zoonosis were reported in Yantai City. The top four diseases reported in incidence rate were severe fever with thrombocytopenia syndrome (SFTS, 3.13/100 000), brucellosis (2.57/100 000), hemorrhagic fever with renal syndrome (HFRS, 1.09/100 000), and scrub typhus (0.73/100 000). There were statistically significant differences in the reported incidence rate of the four major natural focus diseases and vector-borne infectious diseases between different years (χ 2 = 92.55, 34.88, 132.33, 39.95, P < 0.001). The four major natural focal diseases and vector-borne infectious diseases had reported cases throughout the year. SFTS cases peaked in June, accounting for 28.80% (320/1 111). Brucellosis cases were predominantly reported from March to August, accounting for 65.54% (599/914). HFRS cases peaked in November, accounting for 38.66% (150/388), while scrub typhus cases peaked in October, accounting for 69.88% (181/259). The top three districts/cities with the highest incidence of the four major natural focus diseases and vector-borne infectious diseases were Laizhou City (585 cases), Zhaoyuan City (377 cases), and Haiyang City (358 cases). The incidence rates of SFTS in women was higher than that in men (χ 2 = 13.92, P < 0.001), and the incidences rates of brucellosis and HFRS in men were higher than those in women (χ 2 = 123.09, 73.32, P < 0.001). The cases of the four diseases were mainly farmers, accounting for 76.06% (845/1 111), 78.12% (714/914), 81.44% (316/388), and 85.71% (222/259), respectively. Conclusions:From 2019 to 2023, the reported incidence rate of the four major natural focus diseases and vector-borne infectious diseases in Yantai City presents an overall upward trend, with obvious seasonal, regional and demographic characteristics. Targeted prevention and control measures should be developed for high-risk areas and populations based on the characteristics of different diseases.

Objective To investigate the predictive value of preoperative ultrasound in combination with neutrophil-to-lymphocyte ratio(NLR),calcitonin(Ctn),and carcinoembryonic antigen(CEA)for cervical lymph node metastasis in patients with papillary thyroid carcinoma.Methods A total of 103 patients diagnosed with papillary thyroid carcinoma(PTC)who were admitted to the hospital between October 2021 and October 2024 were selected as the case group.Among them,34 patients with cervical lymph node metastasis confirmed by surgical and pathological examination were assigned to the metastatic group,and 69 patients without cervical lymph node metas-tasis were assigned to the non-metastatic group.Additionally,103 patients with benign thyroid nodules admitted during the same period were enrolled as the control group.Clinical data,ultrasonographic features,and serum levels of NLR,Ctn,and CEA were compared between the metastatic and non-metastatic groups.The predictive value of ultrasonographic features and the combined detection of NLR,Ctn,and CEA levels for cervical lymph node metastasis in PTC was evaluated using receiver operating characteristic(ROC)curve analysis.Results Compared to the control group,the case group exhibited a higher proportion of patients with microcalcification and grade 3 blood flow,as well as elevated levels of NLR,Ctn,and CEA(P<0.05).Similarly,compared to the non-metastatic group,the metastatic group showed a higher proportion of patients with microcalcification and grade 3 blood flow,along with increased levels of NLR,Ctn,and CEA(P<0.05).The metastatic group tested positive,whereas the non-metastatic group tested negative.The area under the curve(AUC)for ultrasound features(micro-calcification,blood flow classification)combined with NLR,Ctn,and CEA levels in diagnosing cervical lymph node metastasis in papillary thyroid carcinoma was higher than that of individual indicators(P<0.05).Conclusions Preoperative ultrasound combined with the assessment of NLR,Ctn,and CEA levels demonstrates significant predictive value for cervical lymph node metastasis in papillary thyroid carcinoma.

BACKGROUND:Matrigel is the best material for the culture of tumor organoids,but matrigel alone is not enough to simulate the mechanical environment of tumor growth in vitro.Although the introduction of sodium alginate material can improve the stiffness of the hydrogel based on matrigel,its mechanical properties of hydrogel are difficult to maintain stability in long-term culture.OBJECTIVE:To introduce a small amount of calcium ions into the medium of breast cancer organoids and to observe its maintenance effect on the long-term mechanical properties of the matrigel-sodium alginate hydrogel.METHODS:(1)Sodium alginate composite hydrogels with low,medium,and high stiffness were prepared by introducing different mass concentrations(0,2.5,and 5 mg/mL)of sodium alginate into the constant mass concentration(5 mg/mL)of matrigel.The mechanical properties of hydrogels were measured regularly by rheometer.(2)Human triple negative breast cancer cells MDA-MB-231 were resuspended in hydrogel pre-gels with different stiffness.After gelling,breast cancer organoid factor medium containing(or without)calcium ions was added for breast cancer organoid culture.At a set time point,rheometer was used to regularly measure the effect of calcium ion introduction on the mechanical properties of hydrogel.The morphologic changes of breast cancer organoids were observed under optical microscope.Rate of breast cancer organoids forming into pellets was calculated on day 13.After 7 days of breast cancer organoid culture,different concentrations of the chemotherapy drug docetaxel(0.1,1,10,and 100 nmol/L)were added for intervention for 6 days.Cell viability was detected and the semi-inhibitory concentration of docetaxel,IC50,was calculated.RESULTS AND CONCLUSION:(1)The introduction of sodium alginate effectively improved the mechanical strength of the composite hydrogel.(2)With the extension of breast cancer organoid culture time,the mechanical strength of hydrogels decreased.On day 13 of culture,the mechanical properties of medium and high stiffness hydrogels in the culture environment containing calcium ions were significantly higher than those in the culture environment without calcium ions(P<0.05).There was no significant change in the mechanical properties of low stiffness hydrogels in the two cultures(P>0.05).In long-term culture(13 days),breast cancer organoids changed from round to spindle shape with the decrease of hydrogel mechanical properties in the medium and high stiffness hydrogel groups.After the introduction of calcium ions,the morphology of breast cancer organoids did not change with the extension of culture time in the two groups.The introduction of calcium ions in the culture environment had no effect on the pellet formation rate of breast cancer organoids in the low stiffness hydrogel group,but could improve the pellet formation rate of breast cancer organoids in the medium and high stiffness hydrogel groups.(3)In the culture environment without calcium ions,the cell viability of breast cancer organoids decreased with the increase of docetaxel concentration,and there was no significant difference in IC50 among the three hydrogel groups(P>0.05).In the culture environment containing calcium ions,the cell viability of breast cancer organoids decreased with the increase of docetaxel concentration.The cell viability of breast cancer organoids in the medium and high stiffness hydrogel groups was stronger than that in the low stiffness hydrogel group,and the IC50 was higher than that in the low stiffness hydrogel group(P<0.05).(4)The results showed that the mechanical properties of the matrigel-sodium alginate hydrogel could be maintained by introducing calcium ions into the breast cancer organoid culture system.

Objective:To assess performance advantages of voxel-less optimization(VoLO)algorithm of CyberKnife-based S7 treatment plan system for the optimization of stereotactic body radiation therapy(SBRT)for liver cancer.Methods:The case data of 20 patients with hepatocellular carcinoma from Chinese PLA General Hospital during June 2022 and April 2023 were retrospectively selected,which included 10 patients with large hepatocellular carcinoma and 10 patients with small hepatocellular carcinoma.All patients adopted respectively sequential optimization(SO)and VoLO to conduct optimization for plan.The optimized quality of plan and execution efficiency of two kinds of algorithms were assessed,and the influences of different tumor volumes also were considered.The planed quality assessment included dosimetric parameters of the target region and organ at risk(OAR).The assessment parameters of execution efficiency included the numbers of monitor units(MUs),nodes and beams,and estimated treatment time.Paired t-test method was adopted to analyze quality of plan and treatment efficiency.Results:On the aspect of the dose of target region,for small hepatocellular carcinoma,the conformity index(CI)value(1.08±0.05)of target region of VoLO algorithm was significantly better than(1.17±0.06)of SO algorithm(t=4.631,P<0.05).The gradient index(GI),coverage rate and dose by 95%(D95%)of VoLO algorithm were better than those of SO algorithm,while the differences were not significant(P>0.05).According to the defined standards of liver surgery,for large hepatocellular carcinoma,the differences in CI,GI,coverage rate and D95%of target region between two kinds of algorithms were significant(t=3.337,4.238,-3.359,-3.311,P<0.05),respectively.On the aspect of dosimetry for OAR,for the target region of large hepatocellular carcinoma,the differences of liver Dmean and D700 cm3 between two kinds of algorithms were significant(t=4.114,3.415,P<0.05).However,for small hepatocellular carcinoma,there was no significant statistical difference in dosimetry parameters of OAR between two kinds of algorithms(P>0.05).The execution efficiency of the plan of VoLO group was obviously higher than that of SO group,and the differences of MU number,node number,beam number and estimated treatment time between two groups were significant(t=12.661,4.423,5.024,9.487,P<0.05).Conclusion:The quality of VoLO plan is significantly better than that of SO,which has a significant improvement in execution efficiency of treatment.For the cases of large hepatocellular carcinoma with more complexity,the VoLO optimization shows better advantages on the aspect of dose on target region,and protection for normal liver.

Objective To investigate the effects of galectin-3(Gal3)on autophagy in high glucose-induced human telomerase reverse transcriptase-immortalized retinal pigment epithelial(hTERT-RPE)cells.Methods The hTERT-RPE cells cultured in vitro were randomly divided into a normal group(group C),a high glucose group(group HG),a high glu-cose+si-NC group(group HG+si-NC)and a high glucose+si-Gal3 group(group HG+si-Gal3).Reverse transcrip-tion polymerase chain reaction(RT-PCR)was used to detect the relative mRNA expression levels of Gal3,intercellular ad-hesion molecule(ICAM)-1,interleukin(IL)-6 and tumor necrosis factor(TNF)-α in hTERT-RPE cells of each group.The expression levels of IL-1 and IL-6 in the supernatant of hTERT-RPE cells were measured by enzyme-linked immunosorbent assay(ELISA).The expression level of reactive oxygen species(ROS),the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in hTERT-RPE cells were analyzed by dichlorofluorescin diacetate(DCFH-DA)staining,the colorimetric method and the microplate method,respectively.The protein expression levels of Gal3,the ratio of microtubule associated protein 1 light chain 3(LC3)type Ⅱ to type Ⅰ(LC3 Ⅱ/LC3 Ⅰ),Beclin 1,and autophagy-associat-ed 5(ATG5)and 7(ATG7)in hTERT-RPE cells from each group were detected by Western blot analysis.After the hTERT-RPE cells in each group were transfected with double fluorescent mRFP-GFP-LC3 plasmids alone and with RFP-LAMP1 and GFP-LC3 plasmids jointly,the changes of autophagy flow in hTERT-RPE cells were detected by laser confocal microscopy.Results Compared with the group C,the group HG showed an increase in the expression of Gal3,IL-1,IL-6,TNF-α,ICAM-1,P62,ROS and MDA content(all P＜0.05).However,the expression of LC3 Ⅱ/LC3 Ⅰ,Beclin1,ATG7,ATG5,the SOD activity,and the number of red(autolysosomes)and yellow fluorescence spots(autophagosomes)of the double fluo-rescent mRFP-GFP-LC3 plasmid were all lower in the group HG than those in the group C(all P＜0.05).There was no sig-nificant difference in the number of yellow fluorescent spots(autolysosomes)co-located by RFP-LAMP1 and GFP-LC3 plas-mids between groups C and HG(P＞0.05).There were no significant differences in the expression levels of the above-mentioned indexes in hTERT-RPE cells between groups HG and HG+si-NC(all P＞0.05).The expression levels of Gal3,IL-1,IL-6,TNF-α,ICAM-1,P62,ROS and MDA content in hTERT-RPE cells of the group HG+si-Gal3 were lower than those of the group HG+si-NC(all P＜0.05).Compared with those in the group HG+si-NC,the expression levels of LC3 Ⅱ/LC3 Ⅰ,Beclin1,ATG7,ATG5,SOD activity,and the number of red and yellow fluorescent spots of the double fluo-rescent mRFP-GFP-LC3 plasmid were increased in the group HG+si-Gal3(all P＜0.05).There was no significant differ-ence in the number of yellow fluorescent spots co-located by RFP-LAMP1 and GFP-LC3 plasmids between groups HG+si-GaL3 and HG+si-NC(P＞0.05).Conclusion Gal3 is significantly elevated in hTERT-RPE cells induced by high glu-cose,resulting in impaired autophagy flow.It produces a direct regulatory effect on the formation of autophagosomes,and plays a highly active role in oxidative damage and expression of inflammatory factors in hTERT-RPE cells induced by high glucose.

BACKGROUND:Matrigel is the best material for the culture of tumor organoids,but matrigel alone is not enough to simulate the mechanical environment of tumor growth in vitro.Although the introduction of sodium alginate material can improve the stiffness of the hydrogel based on matrigel,its mechanical properties of hydrogel are difficult to maintain stability in long-term culture.OBJECTIVE:To introduce a small amount of calcium ions into the medium of breast cancer organoids and to observe its maintenance effect on the long-term mechanical properties of the matrigel-sodium alginate hydrogel.METHODS:(1)Sodium alginate composite hydrogels with low,medium,and high stiffness were prepared by introducing different mass concentrations(0,2.5,and 5 mg/mL)of sodium alginate into the constant mass concentration(5 mg/mL)of matrigel.The mechanical properties of hydrogels were measured regularly by rheometer.(2)Human triple negative breast cancer cells MDA-MB-231 were resuspended in hydrogel pre-gels with different stiffness.After gelling,breast cancer organoid factor medium containing(or without)calcium ions was added for breast cancer organoid culture.At a set time point,rheometer was used to regularly measure the effect of calcium ion introduction on the mechanical properties of hydrogel.The morphologic changes of breast cancer organoids were observed under optical microscope.Rate of breast cancer organoids forming into pellets was calculated on day 13.After 7 days of breast cancer organoid culture,different concentrations of the chemotherapy drug docetaxel(0.1,1,10,and 100 nmol/L)were added for intervention for 6 days.Cell viability was detected and the semi-inhibitory concentration of docetaxel,IC50,was calculated.RESULTS AND CONCLUSION:(1)The introduction of sodium alginate effectively improved the mechanical strength of the composite hydrogel.(2)With the extension of breast cancer organoid culture time,the mechanical strength of hydrogels decreased.On day 13 of culture,the mechanical properties of medium and high stiffness hydrogels in the culture environment containing calcium ions were significantly higher than those in the culture environment without calcium ions(P<0.05).There was no significant change in the mechanical properties of low stiffness hydrogels in the two cultures(P>0.05).In long-term culture(13 days),breast cancer organoids changed from round to spindle shape with the decrease of hydrogel mechanical properties in the medium and high stiffness hydrogel groups.After the introduction of calcium ions,the morphology of breast cancer organoids did not change with the extension of culture time in the two groups.The introduction of calcium ions in the culture environment had no effect on the pellet formation rate of breast cancer organoids in the low stiffness hydrogel group,but could improve the pellet formation rate of breast cancer organoids in the medium and high stiffness hydrogel groups.(3)In the culture environment without calcium ions,the cell viability of breast cancer organoids decreased with the increase of docetaxel concentration,and there was no significant difference in IC50 among the three hydrogel groups(P>0.05).In the culture environment containing calcium ions,the cell viability of breast cancer organoids decreased with the increase of docetaxel concentration.The cell viability of breast cancer organoids in the medium and high stiffness hydrogel groups was stronger than that in the low stiffness hydrogel group,and the IC50 was higher than that in the low stiffness hydrogel group(P<0.05).(4)The results showed that the mechanical properties of the matrigel-sodium alginate hydrogel could be maintained by introducing calcium ions into the breast cancer organoid culture system.