Objective To investigate the efficacy and influencing factors of programmed death-1(PD-1)inhibitors in neoadjuvant chemotherapy for triple-negative breast cancer(TNBC).Methods A total of 86 patients with TNBC who received neoadjuvant therapy in The Fifth Medical Center,PLA General Hospital between Jan.1,2018,and Jan.1,2024 and met the inclusion criteria were enrolled,and their clinicopathological data were collected.Based on the neoadjuvant treatment regimens,40 patients who received TP+PD-1 inhibitor(paclitaxel+carboplatin+pembrolizumab)were assigned to TP+PD-1 inhibitor group,and 46 patients who received TP(paclitaxel+carboplatin)were assigned to TP group.The efficacy and incidence of adverse events were compared between the 2 groups after 6 cycles of neoadjuvant therapy.According to the efficacy of neoadjuvant therapy,the patients were further categorized into pathological complete response(pCR)group and non-pCR group.Multivariate logistic stepwise regression analysis was performed to identify independent factors influencing neoadjuvant treatment efficacy.Patients were followed up until Dec.31,2024,and survival analysis was conducted using Kaplan-Meier method.Results There was no significant difference in the objective response rates between the TP+PD-1 inhibitor group and TP group after neoadjuvant therapy(95.0%[38/40]vs 91.3%[42/46],P=0.351].However,the pCR rate was significantly higher in the TP+PD-1 inhibitor group compared with the TP group(65.0%[26/40]vs 43.5%[20/46],P=0.047).There were no significant differences between the 2 groups in terms of disease-free survival,overall survival,or incidence of adverse events(all P＞0.05).Multivariate logistic stepwise regression analysis revealed that the expression of Ki-67 and treatment regimen were influencing factors of pCR after neoadjuvant therapy(odds ratio[OR]=3.382,95%confidence interval[95%CI]1.290-8.868,P=0.013;OR=2.524,95%CI 1.013-6.285,P=0.047).One case of distant metastasis and death occurred in the pCR group,while 8 cases of distant metastasis and 4 deaths occurred in the non-pCR group.The disease-free survival was significantly longer in the pCR group than in the non-pCR group(P=0.031),while the overall survival was similar between the 2 groups(P=0.087).Conclusion Compared with the 6-cycle TP regimen,the 6-cycle TP combined with PD-1 inhibitor regimen can improve the pCR rate in the neoadjuvant treatment of TNBC,with manageable adverse events,suggesting it may serve as a preferred option for TNBC neoadjuvant therapy.Ki-67 expression may serve as a predictive biomarker for achieving pCR.TNBC patients who achieved pCR have better disease-free survival than those who did not.

Lentiviral vectors(LVs)are key gene delivery tools for integrating target genes into the host genome,but they may also pose risks of insertional mutagenesis.The vector copy number(VCN)in cells is critical for determining the safety of gene modification.However,the reliability and accuracy of its quantification process are influenced by multiple factors.Developing cell reference materials with specific vector copy numbers represents a viable approach to enhance the reliability and consistency of measurement results,enabling quality control of the quantification process and traceability of outcomes.However,the preparation of such reference materials faces challenges in cell sample design,preparation protocols,and advanced quantification techniques.In this study,T lymphocyte cell line Jurkat-based reference materials with LV gene copy numbers of 1 and 2 copy/cell were developed.A high-precision duplex digital polymerase chain reaction(dPCR)method was established to quantify the LV gene and endogenous genes simultaneously.Additionally,the results of dPCR were cross-validated through next-generation sequencing and flow cytometric analysis.Ultimately,confocal microscopy characterization results showed that the developed cell reference materials had intact morphology.The quantification result of VCN-1 was(1.07±0.11)copy/cell,and that of VCN-2 was(2.09±0.21)copy/cell.These cell reference materials demonstrated compliance with stability and homogeneity requirements,and could be applied for quality control throughout the VCN measurement workflow and metrological traceability,improving the accuracy,comparability,and validity of copy number measurements.

ObjectiveTo explore the possible mechanism of Chuanxiong （Rhizoma Chuanxiong） & Tianma （Rhizoma Gastrodiae） herbal pair in treating migraines based on AMP-activated protein kinase （AMPK）/transient receptor potential A1 channel （TRPA1） pathway. MethodsForty-eight healthy male SD rats were randomly divided into control group， model group， and Chuanxiong Tianma medication group， with 16 rats in each group. The control group and model group were given 10 ml/kg of normal saline by gavage， while the Chuanxiong Tianma medication group was given 0.675 g/kg of Chuanxiong Tianma herbal pair by gavage， once daily for 8 consecutive days in both groups. Migraime model was performed before the last administration， with subcutaneous injection of 10 ml/kg of normal saline in the control group， and subcutaneous injection of 10 ml/kg of nitroglycerin in the model group and Chuanxiong Tianma medication group. The Von Frey filament was used to measure the periorbital mechanical pain threshold of rats. The enzyme-linked immunosorbent assay （ELISA） was used to determine the levels of calcitonin gene-related peptide （CGRP） in rat serum and cerebrospinal fluid. The nitric oxide （NO） assay kit was used to determine the NO level in serum and cerebrospinal fluid. RT-PCR was usedto detect the mRNA expression levels of immediate-early genes in the trigeminal ganglion of rats （c-Fos）， CGRP， transient receptor potential V1 channel （TRPV1）， AMPK alpha subunit （PRKAA）， and TRPA1. Immunofluorescence was used to detect the number of c-Fos-positive cells in the trigeminal cervical complex （TCC） and the protein expression levels of phosphorylated AMPK （pAMPK） and TRPA1 in the trigeminal ganglion. ResultsCompared to those in the control group， the mechanical stimulation threshold and pAMPK protein expression in the model group decreased， while the levels of CGRP and NO in serum， c-Fos， CGRP， TRPV1 and TRPA1 mRNA levels in the trigeminal ganglion， TRPA1 protein expression， and the number of c-Fos-positive cells in the TCC significantly increased （P＜0.05）. Compared to those in the model group， the mechanical stimulation threshold and pAMPK protein expression in the Chuanxiong Tianma medication group significantly increased， while the levels of CGRP and NO in serum， c-Fos， CGRP， TRPV1 and TRPA1 mRNA levels in the trigeminal ganglion， TRPA1 protein expression， and the number of c-Fos-positive cells in the TCC significantly decreased （P＜0.05）. ConclusionChuanxiong Tianma herbal pair may improve migraine symptoms by regulating the AMPK/TRPA1 pathway in the trigeminal ganglion and increasing the mechanical pain threshold.

Objective: To analyze the trends of overweight and obesity in children and adolescents aged 7-18 in Hainan Province from 2005 to 2019, in order to provide a scientific basis for prevention strategies. Methods: Data were sourced from a total of 42 811 students in Hainan Province in 2005, 2010, 2014 and 2019 for physical fitness and health investigation, according to the criteria of Screening for Overweight and Obesity among School aged Children and Adolescents. The linear trend of overweight and obesity was compared by using the χ 2 test and χ 2 trend test, and with 2005 as the base period, the dynamic analysis was conducted to compare the growth and development rates of overweight and obesity. Results: The detection rates of overweight in children and adolescents aged 7-18 in Hainan Province were 4.7%, 5.1%, 6.7% and 10.8% in 2005, 2010, 2014 and 2019, respectively, while the detection rates of obesity were 2.7%, 2.4%, 2.5% and 6.8%, which showed an overall upward trend by year ( χ 2 trend =370.57 , 313.55, P <0.01). All differences in overweight and obesity detection rates across gender, ethnicity, and age groups were statistically significant for each year ( χ 2=9.56-290.95, P <0.01). And the detection rates of overweight and obesity increased linearly over year in different characteristic groups ( χ 2 trend =29.92-280.11, P <0.01). The annual growth rate of overweight detection rate was higher for girls (6.4%) than for boys (6.1%), higher for ethnic minorities (7.1%) than for Han (5.7%), and higher for the 16- 18 age group (8.4%) than for the 7-9 (3.6%), 10-12 (6.2%), and 13-15 age groups (7.7%). And the annual growth rate of obesity detection rate was higher for boys (8.1%) than for girls ( 5.7% ), higher for ethnic minorities (7.1%) than for Han ( 6.8% ), and higher for the 16-18 age group (77.2%) than for the 7-9 (2.8%), 10-12 (8.2%), and 13-15 (18.7%) age groups. Conclusions The overall prevalence of overweight and obesity among children and adolescents aged 7-18 in Hainan Province continues to increase from 2005 to 2019, with differences in genders, ethnic and ages. Interventions should be implemented to reduce the incidence of overweight obesity in children and adolescents.

Objective: To understand the trends in malnutrition among primary and middle school students of Han nationality in Hainan Province from 2005 to 2019, so as to provide a basis for improving nutrition intervention measures for children and adolescents. Methods: A sample of 32 949 Han nationality primary and middle school students aged 7-18 years old in Hainan Province were investigated in 2005, 2010, 2014 and 2019 based on national survey on student physical fitness and health. The Malnutrition Screening Standard of Schoolage Children and Adolescents was used to screen malnutrition. Statistical analysis was performed using the χ2 test and the χ2trend test. Results: In the four surveys conducted during 2005 to 2019, the prevalence of malnutrition among primary and middle school students were 22.12%, 18.80%, 15.89% and 9.56%, respectively, with an increase of -12.56% and an average annual increase of -5.82%. The decreasing trend of malnutrition by year was statistically significant (χ2trend=600.72, P<0.01), and the proportion of emaciation type was the highest (8.87%-20.15%). The detection rates of malnutrition among all students aged 7 to 18 showed a decreasing trend from 2005 to 2019 (χ2trend=56.44, 60.04, 61.48, 42.49, 51.81, 50.81, 72.86, 101.34, 86.38, 24.81, 17.72, 10.38, P<0.01). From 2005 to 2019, the detection rates of malnutrition in boys were higher than that of girls (in 4 surveys), and that in rural students from 2005 to 2014 of 3 surveys were higher than that in towns (χ2=92.07, 35.16, 25.29, 29.98; 64.35, 4.26, 6.32, P<0.05). Conclusions The malnutrition of Han nationality primary and middle school students aged 7-18 years in Hainan Province show a trend of improvement year by year from 2005 to 2019, despite the overall high detection rate. Wasting is the most common type of malnutrition. The epidemic of malnutrition varies by age, sex and areas. Further targeted measures should be taken to strengthen intervention in the diet of primary and middle school students, to improve the nutritional status of children and adolescents.

Objective To investigate the risk factors of in-hospital mortality and establish a risk prediction model for patients receiving venoarterial extracorporeal membrane oxygenation(VA-ECMO).Methods We retrospectively collected the data of 302 patients receiving VA-ECMO in ICU of 3 hospitals in Guangdong Province between January,2015 and January,2022 using a convenience sampling method.The patients were divided into a derivation cohort(201 cases)and a validation cohort(101 cases).Univariate and multivariate logistic regression analyses were used to analyze the risk factors for in-hospital death of these patients,based on which a risk prediction model was established in the form of a nomogram.The receiver operator characteristic(ROC)curve,calibration curve and clinical decision curve were used to evaluate the discrimination ability,calibration and clinical validity of this model.Results The in-hospital mortality risk prediction model was established based the risk factors including hypertension(OR=3.694,95%CI:1.582-8.621),continuous renal replacement therapy(OR=9.661,95%CI:4.103-22.745),elevated Na2+ level(OR=1.048,95%CI:1.003-1.095)and increased hemoglobin level(OR=0.987,95%CI:0.977-0.998).In the derivation cohort,the area under the ROC curve(AUC)of this model was 0.829(95%CI:0.770-0.889),greater than those of the 4 single factors(all AUC<0.800),APACHE Ⅱ Score(AUC=0.777,95%CI:0.714-0.840)and the SOFA Score(AUC=0.721,95%CI:0.647-0.796).The results of internal validation showed that the AUC of the model was 0.774(95%CI:0.679-0.869),and the goodness of fit test showed a good fitting of this model(χ2=4.629,P>0.05).Conclusion The risk prediction model for in-hospital mortality of patients on VA-ECMO has good differentiation,calibration and clinical effectiveness and outperforms the commonly used disease severity scoring system,and thus can be used for assessing disease severity and prognostic risk level in critically ill patients.

Objective To explore the mechanism of quercetin in the treatment of breast cancer with depressive features using network pharmacology and animal experiments.Methods Network pharmacology and bioinformatics methods were used to predict the key targets and mechanisms of quercetin in the treatment of breast cancer with depressive characteristics.The predicted results were verified by animal experiments.A mouse model of breast cancer with depressive characteristics was constructed,and quercetin intervention was performed after grouping.The depression of mice was evaluated by open field test.The tumor volume and tumor mass were measured.The expression of Ki-67 in tumor tissue was detected by immunohistochemical staining.The expressions of tumor necrosis factor α(TNF-α),interleukin 6(IL-6),p53,Caspase-3 and B-cell lymphoma/leukemia 2(Bcl-2)in tumor tissue were detected by Western Blot.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)method was used to detect the apoptosis of tumor cells.Results In the breast cancer model group with depressive characteristics,the total movement distance in the open field test and the ratio of residence time in the central area of the open field test were decreased,the tumor volume and tumor mass were significantly increased,and Ki-67 expression level in the tumor tissue was significantly increased,the expression levels of TNF-α,IL-6,p53 and Caspase-3 in the tumor tissue were decreased and the expression level of Bcl-2 was increased,as well as the rate of TUNEL positive cells was decreased,the differences being statistically significant compared with the control group(P＜0.01 or P＜0.001).Compared with the model group,the above indexes were significantly reversed in the quercetin group(P＜0.01 or P＜0.001).Conclusion Quercetin can effectively inhibit the progression of breast cancer with depressive characteristics in mice,and its mechanism is related to the regulation of TNF,IL6,TP53,CASP3,BCL2 and other targets to promote tumor cell apoptosis.

Objective:To analyze and explore the effects of Huayu Jiedu Decoction on the malignant biological characteristics of multiple myeloma(MM)cells and its molecular mechanism,so as to provide experimental basis and theoretical basis for the alternative therapy of anti-MM in traditional Chinese medicine.Methods:Different concentrations of Huayu Jiedu Decoction were used to intervene myeloma U266 cells.The changes of cell proliferation activity were detected by CCK-8 assay,apoptosis was detected by Annexin V/PI double staining flow cytometry,and apoptosis and protein expression of related signaling pathways were detected by Western blot.Real-time quantitative PCR was used to detect mRNA expression changes of high mobility group protein B1(HMGB1),CXC chemokine receptor 4(CXCR4)and interleukin-6(IL-6).Results:Huayu Jiedu Decoction inhibited the proliferative activity of U266 cells and induced their apoptosis in a concentration and time dependent manner(r=-0.713,r=-0.827).After treatment with Huayu Jiedu Decoction for 48 h,the expressions of anti-apoptotic protein Bcl-2 and survivin were down-regulated,while the expression of pro-apoptotic protein Bax was up-regulated,and the phosphorylation level of TLR4/NF-κB signaling pathway was inhibited.After intervention of Huayu Jiedu decoction,the expressions of HMGB1 and IL-6 mRNA were significantly decreased,while the expression of CXCR4 was not significantly decreased.Conclusion:Huayu Jiedu Decoction can inhibit the proliferative activity of U266 cells and induce programmed death.Its molecular mechanism may be related to regulating the expression of apoptotic proteins,inhibiting the activation of TLR4/NF-κB pathway and down-regulating the expression of HMGB1 and IL-6 mRNA.

Objective:To investigate the effects of Curcumol on the malignant biological characteristics of acute myeloid leukemia (AML)cells and its molecular mechanism,and to provide theoretical and experimental evidence for the anti-leukemia treatment of traditional Chinese medicine.Methods:After the AML cell lines HL-60 and KG-1 cells were treated different concentrations of with Curcumol.The proliferation activity of cells was detected by CCK-8 method,and the expression changes of apoptotic proteins and PI3 K/AKT signaling pathway proteins were detected by Western blot. Real-time quantitative fluorescence polymerase chain reaction (RT-qPCR ) was used to detect the expression of Caspase family mRNA.Results:Curcumol could inhibit the proliferation and induce apoptosis of HL-60 and KG-1 cells,promote apoptosis by up-regulating the expression of Bax and down-regulating the expression of Bcl-2 protein (P<0.05).When Curcumol interferes with HL-60 and KG-1 cells,it can also induce programmed cell death of AML by inhibiting PI3 K/AKT signaling pathway.In addition,after the intervention of Curcumol,the expression of Caspase 3,Caspase 6,Caspase 8 and Caspase 9 were up-regulated in HL-60 cells (P<0.05 ),the expression of Caspase 3,Caspase 8 and Caspase 9 were significantly up-regulated in KG-1 cells (P<0.01),while the expression of Caspase 6 was weakly affected (P<0.05 ),but low concentration of Curcumol (<60 μg/ml)had no effect on the expression of Caspase 6 in KG-1 cells (P>0.05).Conclusion:Curcumol may mediate the programmed death of AML cells by inhibiting the PI3K/AKT signaling pathway,affecting the expression of Bcl-2 family proteins,and promoting the activation of core members of Caspase family,so as to play an anti-leukemia role.

Objective To investigate the risk factors of in-hospital mortality and establish a risk prediction model for patients receiving venoarterial extracorporeal membrane oxygenation(VA-ECMO).Methods We retrospectively collected the data of 302 patients receiving VA-ECMO in ICU of 3 hospitals in Guangdong Province between January,2015 and January,2022 using a convenience sampling method.The patients were divided into a derivation cohort(201 cases)and a validation cohort(101 cases).Univariate and multivariate logistic regression analyses were used to analyze the risk factors for in-hospital death of these patients,based on which a risk prediction model was established in the form of a nomogram.The receiver operator characteristic(ROC)curve,calibration curve and clinical decision curve were used to evaluate the discrimination ability,calibration and clinical validity of this model.Results The in-hospital mortality risk prediction model was established based the risk factors including hypertension(OR=3.694,95%CI:1.582-8.621),continuous renal replacement therapy(OR=9.661,95%CI:4.103-22.745),elevated Na2+ level(OR=1.048,95%CI:1.003-1.095)and increased hemoglobin level(OR=0.987,95%CI:0.977-0.998).In the derivation cohort,the area under the ROC curve(AUC)of this model was 0.829(95%CI:0.770-0.889),greater than those of the 4 single factors(all AUC<0.800),APACHE Ⅱ Score(AUC=0.777,95%CI:0.714-0.840)and the SOFA Score(AUC=0.721,95%CI:0.647-0.796).The results of internal validation showed that the AUC of the model was 0.774(95%CI:0.679-0.869),and the goodness of fit test showed a good fitting of this model(χ2=4.629,P>0.05).Conclusion The risk prediction model for in-hospital mortality of patients on VA-ECMO has good differentiation,calibration and clinical effectiveness and outperforms the commonly used disease severity scoring system,and thus can be used for assessing disease severity and prognostic risk level in critically ill patients.