Objective Triple-negative breast cancer(TNBC)poses a significant challenge for treatment efficacy.CD8+T cells,which are pivotal immune cells,can be effectively analyzed for differential gene expression across diverse cell populations owing to rapid advancements in sequencing technology.By leveraging these genes,our objective was to develop a prognostic model that accurately predicts the prognosis of patients with TNBC and their responsiveness to immunotherapy. Methods Sample information and clinical data of TNBC were sourced from The Cancer Genome Atlas and METABRIC databases.In the initial stage,we identified 67 differentially expressed genes associated with immune response in CD8+T cells.Subsequently,we narrowed our focus to three key genes,namely CXCL13,GBP2,and GZMB,which were used to construct a prognostic model.The accuracy of the model was assessed using the validation set data and receiver operating characteristic(ROC)curves.Furthermore,we employed various methods,including Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway,immune infiltration,and correlation analyses with CD274(PD-L1)to explore the model's predictive efficacy in immunotherapeutic responses.Additionally,we investigated the potential underlying biological pathways that contribute to divergent treatment responses. Results We successfully developed a model capable of predicting the prognosis of patients with TNBC.The areas under the curve(AUC)values for the 1-,3-,and 5-year survival predictions were 0.618,0.652,and 0.826,respectively.Employing this risk model,we stratified the samples into high-and low-risk groups.Through KEGG enrichment analysis,we observed that the high-risk group predominantly exhibited enrichment in metabolism-related pathways such as drug and chlorophyll metabolism,whereas the low-risk group demonstrated significant enrichment in cytokine pathways.Furthermore,immune landscape analysis revealed noteworthy variations between(PD-L1)expression and risk scores, Conclusion Our study demonstrates the potential of CXCL13,GBP2,and GZMB as prognostic indicators of clinical outcomes and immunotherapy responses in patients with TNBC.These findings provide valuable insights and novel avenues for developing immunotherapeutic approaches targeting TNBC.

Objective:To investigate the efficacy and safety of chemotherapy combined with inetetamab and pyrotinib (triplet therapy group), and capecitabine combined with pyrotinib (doublet therapy group) in second-line treatment for HER2-positive metastatic breast cancer patients who have failed on trastuzumab.Methods:A retrospective analysis was conducted on 30 HER2-positive metastatic breast cancer patients who underwent second-line treatment with a regimen of chemotherapy plus inetetamab and pyrotinib at Fujian Provincial Tumor Hospital between Jan. 1, 2020 and Dec. 31, 2023, and on 28 HER2-positive metastatic breast cancer patients treated with capecitabine plus pyrotinib during the same period. The study analyzed the differences in ORR (objective response rate), CBR (clinical benefit rate), PFS (progression free survival), and OS (overall survival) between the two groups and documented any adverse reactions observed during treatment.Results:The three-drug group exhibited a higher ORR rate than the two-drug group (60.0% vs. 39.2%, P>0.05), and a higher CBR rate (83.3% vs. 57.1%, P<0.05). Univariate analysis and multivariate Logistic regression analysis showed that group (three drug group compared to two drug group) was the only independent prognostic factor affecting median PFS in patients ( P<0.05). The median PFS of the three drug group was higher than that of the two drug group (19 months vs. 11 months, P<0.05), but by the end of follow-up, neither group had reached the median OS ( P>0.05) .The three-drug group and the two-drug group demonstrated 1-year OS rates of 85% and 85%, 2-year OS rates of 74% and 63%, and 3-year OS rates of 56% and 52%, respectively. The most frequent hematological toxicities in the three-drug group were leukopenia, neutropenia, and anemia, with diarrhea being the most common non-hematological toxicity. No significant differences were observed in adverse reaction profiles between the two groups ( P>0.05) . Conclusion:In the second-line treatment of HER2 positive metastatic breast cancer, chemotherapy + inetetamab + pyrotinib significantly prolonged the median PFS and increased the CBR compared with the current domestic standard capecitabine + pyrotinib regimen, and did not increase the incidence of adverse reactions, demonstrating higher efficacy and safety.

Objective:To investigate the efficacy and safety of chemotherapy combined with inetetamab and pyrotinib (triplet therapy group), and capecitabine combined with pyrotinib (doublet therapy group) in second-line treatment for HER2-positive metastatic breast cancer patients who have failed on trastuzumab.Methods:A retrospective analysis was conducted on 30 HER2-positive metastatic breast cancer patients who underwent second-line treatment with a regimen of chemotherapy plus inetetamab and pyrotinib at Fujian Provincial Tumor Hospital between Jan. 1, 2020 and Dec. 31, 2023, and on 28 HER2-positive metastatic breast cancer patients treated with capecitabine plus pyrotinib during the same period. The study analyzed the differences in ORR (objective response rate), CBR (clinical benefit rate), PFS (progression free survival), and OS (overall survival) between the two groups and documented any adverse reactions observed during treatment.Results:The three-drug group exhibited a higher ORR rate than the two-drug group (60.0% vs. 39.2%, P>0.05), and a higher CBR rate (83.3% vs. 57.1%, P<0.05). Univariate analysis and multivariate Logistic regression analysis showed that group (three drug group compared to two drug group) was the only independent prognostic factor affecting median PFS in patients ( P<0.05). The median PFS of the three drug group was higher than that of the two drug group (19 months vs. 11 months, P<0.05), but by the end of follow-up, neither group had reached the median OS ( P>0.05) .The three-drug group and the two-drug group demonstrated 1-year OS rates of 85% and 85%, 2-year OS rates of 74% and 63%, and 3-year OS rates of 56% and 52%, respectively. The most frequent hematological toxicities in the three-drug group were leukopenia, neutropenia, and anemia, with diarrhea being the most common non-hematological toxicity. No significant differences were observed in adverse reaction profiles between the two groups ( P>0.05) . Conclusion:In the second-line treatment of HER2 positive metastatic breast cancer, chemotherapy + inetetamab + pyrotinib significantly prolonged the median PFS and increased the CBR compared with the current domestic standard capecitabine + pyrotinib regimen, and did not increase the incidence of adverse reactions, demonstrating higher efficacy and safety.

Objective : To investigate the serum levels of antibodies against SARS-CoV-2 after inoculation of an inactivated SARS-CoV-2 vaccine, so as to provide insights into the evaluation of the vaccine immunogenicity. Methods : In this single-arm Objective performance criteria trial, residents aged 18 to 59 years and inoculated with an inactivated SARS-CoV-2 vaccine in Xihu District, Hangzhou City from October to December of 2020 were selected using a cluster sampling method. Blood samples were collected prior to inoculation, 14 and 28 days post-inoculation of the first dose, and 28 days post-inoculation of the second dose. Serum levels of anti-SARS-CoV-2 IgM and IgG antibodies were detected using the magnetic particle-based chemiluminescence immunoassay. The seroconversion of antibodies and dynamic changes of antibody levels were analyzed. Results : Totally 310 participants were enrolled, including 133 subjects on day 14 post-inoculation of the first dose, 97 subjects on day 28 post-inoculation of the first dose and 254 subjects on day 28 post-inoculation of the second dose. The seroconversion rates of anti-SARS-CoV-2 IgG antibody were 6.02%, 28.87% and 98.43%, and the median IgG antibody levels were 1.76 ( interquartile range, 3.25 ), 5.69 ( 9.95 ) and 52.05 ( 47.60 ) AU/mL ( P<0.05 ), respectively, while the seroconversion rates of anti-SARS-CoV-2 IgM antibody were 9.02%, 11.34% and 12.99%, and the median IgG antibody levels were 1.89 ( 3.28 ), 2.06 ( 4.71 ) and 2.65 ( 4.01 ) AU/mL ( P>0.05 ), respectively. In addition, higher serum levels of anti-SARS-CoV-2 IgG and IgM antibodies were detected post-inoculation relative to pre-inoculation ( P<0.05 ), and higher serum IgG antibody levels were found in subjects aged 18 to 39 years than in those aged 40 to 59 years ( P<0.05 ). Conclusions Inoculation of two doses of the inactivated SARS-CoV-2 vaccine achieves a high immunogenicity among residents aged 18 to 59 years 28 days post-inoculation, and the anti-SARS-CoV-2 IgM antibody is detectable in some residents following inoculation of the first dose.

Objective:To detect the serum levels of SARS-CoV-2-specific IgM and IgG antibodies in patients infected with SARS-CoV-2 and recipients of inactivated vaccine in different periods for understanding their variation patterns in vivo. Methods:Chemiluminescence immunoassay was used to detect the levels of SARS-CoV-2-specific IgM and IgG antibodies in 144 serum samples of 44 COVID-19 patients, 381 serum samples of 118 asymptomatic infected cases and 398 serum samples of 273 inactivated vaccine recipients collected at different periods. The results were statistically analyzed together with basic characteristics and vaccination status.Results:The positive rates of IgM antibody in COVID-19 patients, asymptomatic infected cases and inactivated vaccine recipients were 52.27% (23/44), 23.73% (28/118) and 14.29% (39/273). The positive rate of IgM antibody was higher in COVID-19 patients than in asymptomatic infected cases and vaccine recipients (χ 2=12.106, P=0.001; χ 2=34.755, P<0.001). The positive rates of IgG antibody in the three populations were 100.00% (44/44), 97.46% (115/118) and 98.81% (166/168), and the differences were not statistically significant (χ 2=2.944, P=0.229). In COVID-19 patients, the concentration of IgM antibody in <40 years old group was lower than that in ≥40 years old group (Waldχ 2=6.609, P=0.010), and the concentration of IgG antibody in patients with vaccination was higher than that in patients without vaccination (Waldχ 2=12.402, P<0.001). In asymptomatic infected cases, the concentration of IgG antibody was higher in people with vaccination than in those without vaccination (Waldχ 2=4.530, P=0.033). In SARS-CoV-2 vaccine recipients, the concentration of IgG antibody in <40 years old group was higher than that in ≥40 years old group (Waldχ 2=9.565, P=0.002). Dynamic analysis of antibody levels showed that from week 1 to week 9, the concentrations of IgM and IgG antibodies in COVID-19 patients were higher than those in asymptomatic infected cases and vaccine recipients. Conclusions:The concentrations of IgM and IgG antibodies in COVID-19 patients were higher than those in asymptomatic infected cases and inactivated vaccine recipients. COVID-19 patients aged ≥40 years had higher level of IgM antibody. COVID-19 patients and asymptomatic infected cases who had received vaccination had higher concentration of IgG antibody. Inactivated vaccine showed good immunogenicity after whole course of immunization, and the IgG antibody level in <40 years old group was higher.

OBJECTIVE： To establish a method for simultaneous determination of 4 triglyceride anti-tumor components in Coix lacryma seed oil. METHODS： HPLC-ELSD was adopted. The determination was performed on Inertsil ODS-3 C18 column with mobile phase consisted of acetonitrile-isopropanol （57 ∶ 43， V/V） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃， and the sample size was 10 μL. Evaporative light scattering detector was used， the drift tube temperature was 70 ℃， and the gas flow rate was 2 L/min. Using glycerol trioleateas internal standard， relative correction factors （RCF） of linolein trilinolein， 1，2-linoleic acid-3-palmitic acid glyceride and 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride were calculated respectively. The contents of above 3 components in C. lacryma seed oil were calculated by RCF. The contents of 4 components in C. lacryma seed oil were determined by external standard. The results of content determination by quatitative analysis of multi-components by single marker （QAMS） were compared with external standard method. RESULTS： The linear ranges were 0.15-4.50 μg for linolein trilinolein， 0.15-4.50 μg for 1，2-linoleic acid-3-palmitate， 0.35-10.50 μg for 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride， 0.35-10.50 μg for glycerol trioleate （r≥0.999 5）. The limits of quantification were 0.13， 0.06， 0.07， 0.12 μg. The limits of detection were 0.04， 0.02， 0.02， 0.03 μg， respectively. RSDs of precision， stability， and repeatability tests were less than 2.0％（n＝6）. The average recoveries were 95.43％-102.67％（RSD＜2.0％， n＝6）. Average RCFs of linolein trilinolein， 1，2- linoleic acid-3-palmitic acid glyceride and 1-palmitic acid-2- oleic acid-3-linoleic acid glyceride were 0.31， 0.88， and 1.21， respectively. RCFs reproducibility was perfect under different experiment conditions. There was no significant difference in results of content determination between QAMS and external standard method （P＞0.05）. CONCLUSIONS： The method is simple， rapid， accurate and reliable. It is used for simultaneous determination of linolein trilinolein， 1， 2-linoleic acid-3-palmitate， 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride and glycerol trioleateas in C. lacryma seed oil.

A new analytical method has been developed for the simultaneous determination of CrⅢand CrⅥusing on-line sample pretreatment valve-switching ion chromatography. The organic matrix in leather was removed by using a reverse-phase column as the pretreatment column. Before injection, EDTA was added into sample solution to react with the CrⅢto form anion which could absorb visible light strongly. After injection, the ions separated by the pretreatment column were received in a collection loop. Then the ions were delivered into an analytical column and separated. CrⅥ then was derived with the derivatization reagent 1, 5-diphenylcarbazide ( DPC) , and detected together with CrⅢ-EDTA complex by a UV-Vis detector. Under the optimum conditions, the linear range of the method for CrⅢ and CrⅥ was 0. 3-10 mg/L (r=0. 9991) and 0. 05-2 mg/L ( r = 0. 9992 ), whereas detection limits ( S/N = 3 ) were 80. 78 μg/L and 6. 67 μg/L, respectively. The recoveries were in the range of 88. 7%-108. 5% with the relative standard deviations for retention time and peak area less than 3%. The method could be applied to determine CrⅢ and CrⅥ in leather and cloth effectively and quickly.

OBJECTIVETo establish homozygous transgenic mouse strain expressing human tau isoform with P301L mutation.

METHODSFive transgenic mice expressing human tau isoform with P301L mutation were obtained by microinjection into male nuclei. Homozygote and hemizygote were identified by PCR and real-time fluorescent quantitative PCR.

RESULTSNinety five homozygous transgenic mice were selected, and the results indicated that homozygous transgenic mice were superior to hemizygote in simulating the changes of biological characteristics.

CONCLUSIONExogenous gene tau is able to stably transmit to next generation and the combination of SYBR Green real-time fluorescent quantitative PCR with the traditional mating is a fast, reliable and economical way to screen homozygous and hemizygous transgenic mice.

Animals ; Female ; Homozygote ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microinjections ; Mutation ; tau Proteins ; genetics