Development and validation of a nomogram for predicting lymph node metastasis in ductal carcinoma in situ with microinvasion: A SEER population-based study.

BACKGROUND: Ductal carcinoma in situ with microinvasion (DCIS-MI) is a special type of breast cancer. It is an invasive lesion less than 1.0 mm in size related to simple ductal carcinoma in situ (DCIS). Lymph node metastasis (LNM) in DCIS-MI often indicates a poor prognosis. Therefore, the management of lymph nodes plays a vital role in the treatment strategy of DCIS-MI. Since DCIS-MI is often diagnosed by postoperative paraffin section and immunohistochemical detection, to obtain the best clinical benefits for such patients, we aim to establish and verify a nomogram to predict the possibility of lymph node metastasis in DCIS-MI patients and help preoperative or intraoperative clinical decision-making. METHODS: A retrospective analysis of patients with DCIS-MI in the Surveillance, Epidemiology, and End Results (SEER) database from 2010 to 2019 was performed. The study cohort was randomly divided into a training cohort and a validation cohort at a ratio of 7:3. The risk factors were determined by univariate and multivariate logistic regression analyses in the training cohort, and a nomogram was constructed. The receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis (DCA) were used to evaluate the nomogram in the training set and validation set. An independent data cohort was obtained from the Shanghai Jiao Tong University Breast Cancer Database (SJTU-BCDB) for external validation. RESULTS: This study included 3951 female patients from SEER with DCIS-MI, including 244 patients with regional lymph node metastasis, accounting for 6.18% of the total. An independent test set of 323 patients from SJTU-BCDB was used for external validation. According to the multifactorial logistic regression analysis results, age at diagnosis, ethnicity, grade, and surgical modality were included in the prediction model. The areas under the ROC curves (AUCs) were 0.739 (95% CI: 0.702~0.775), 0.732 (95% CI: 0.675~0.788), and 0.707 (95%CI: 0.607-0.807) in the training, validation and external test groups, suggesting that the column line graphs had excellent differentiation. The calibration curves slope was close to 1, and the model's predicted values were in good agreement with the actual values. The DCA curves showed good clinical utility. CONCLUSION: In this study, we constructed accurate and practical columnar maps with some clinical benefit to predict the likelihood of lymph node metastasis in patients with postoperatively diagnosed DCIS-MI and provide a reference value for specifying treatment strategies.

[Anti-gastric cancer effects of Z Ajoene and its molecular mechanisms].

Objective: To investigate the effects of Z Ajoene on gastric cancer cell MGC-803 and its molecular mechanisms. Methods: The gastric cancer cells MGC-803 were treated with 0, 1, 5, 25 and 125 µmol/L Z Ajoene for 24 h, 48 h and 72 h, each with 3 replicate wells. The proliferation activity of MGC-803 cells was analyzed by MTS method, mitochondrial membrane potential was analyzed after JC-1 staining, nuclear type was observed after Hoechst 33342 staining, cytotoxicity was detected by LDH release method, and the apoptosis level and cell cycle were analyzed with flow cytometry. RT-qPCR and Western blot methods were used to evaluate the expression levels of P53, Caspase-3, RAS, ERK, BCL-2, AKT, mTOR and PI3K genes. At the same time, 4-week-old male BALB/C mice were randomly divided into 5 groups, 20 per group, and were subcutaneously inoculated with gastric cancer cell MGC-803 in the groin. Two days later, each group was injected with Z Ajoene at the doses of 0, 1, 5, 25 and 125 µmol/L, 0.1 ml/time, and was injected every other day. On the 20th day of the first injection of tumor cells, 10 mice in each group were killed, the tumor tissues were taken out and weighed. The survival period of the remaining mice was recorded and the effects of Z Ajoene on the growth and survival period of gastric cancer in tumor-bearing mice were observed. Results: After Z Ajoene treatment, the proliferation activity of MGC-803 cells was significantly inhibited and the apoptosis rate was significantly increased(P＜0.01). The transcription and expression levels of p53, Caspase-3 and BAX genes were significantly increased, while the transcription and expression levels of RAS, ERK1, BCL-2, AKT, mTOR and PI3K genes were decreased markedly(P＜0.01). The tumor inhibition experiments showed that the growth of the tumor could be inhibited and the survival time of the tumor-bearing animals could be greatly prolonged after Z Ajoene treatment(P＜0.01). Conclusion: Z Ajoene has therapeutic effects on gastric cancer, can inhibit the proliferation of gastric cancer cells and induce them apoptosis by regulating the expression of PI3K-AKT-mTOR and RAS-RAF-MEK-ERK signal pathways.

Acute hepatotoxicity of multimodal targeted imaging contrast agent NaLuF4:Gd,Yb,Er-PEG/PEI-FA in mice.

In the past few decades, upconversion nanoparticles (abbreviated as UCNPs) have been more widely applied in the biomedical fields, such as in vitro and in vivo upconversion fluorescent bioimaging, photodynamic therapy, biological macromolecular detection, imaging mediated drug delivery and so on. But meanwhile, there is still not much research on the acute toxicity of upconversion nanoparticles in vivo, such as acute hepatotoxicity. In this work, we studied the in vivo biodistribution and acute hepatotoxicity of multimodal targeted contrast agent NaLuF4:Gd,Yb,Er-PEG/PEI-FA nanoprobe, which were synthesized by the solvothermal method and modified with Polyethylene glycol (PEG), Polyetherimide (PEI), folic acid (FA) on the surface. The acute hepatotoxicity in mice was systematically assessed after tail vein injection of different concentration of UCNPs. The results showed that NaLuF4:Gd,Yb,Er-PEG/PEI-FA nanoparticles with an average diameter of 44.5 ± 10.4 nm, and three typical upconversion fluorescence emission bands at 520 nm, 540 nm and 660 nm under the excitation of 980 nm laser. In vivo distribution experiments results demonstrated that approximately 87% of UCNPs injected through the tail vein accumulate in the liver. In the acute hepatotoxicity test, the intravenously injection dose of UCNPs was 10, 40, 70 and 100 mg/kg, respectively. The body weight, blood routine, serum biochemistry, histomorphology and liver oxidative stress were detected and observed no significant acute hepatotoxicity damage under the injection dose of 100 mg/kg. In conclusion, NaLuF4:Gd,Yb,Er-PEG/PEI-FA nanoprobes are safe and reliable, and have potential applications in the field of tumor targeted multimodal imaging.

[Anti-hepatoma effects of Smac analogue Birinapant and its related molecular mechanism].

OBJECTIVE: To investigate the effects of Birinapant on hepatocellular carcinoma cells and its related molecular mechanisms. METHODS: Human hepatocellular carcinoma cells QGY-7701 were treated with 0, 1, 5, 25 and 125 nmol/L Birinapant for 24, 48 and 72 hours respectively, each experiment 3 wells.The proliferation activity of cells, the apoptosis levels, the cells nuclear type, the mitochondrial membrane potential, the transcription and expression levels of genes and the cytotoxicity of Birinapant were analyzed.At the same time, 4-week-old male BALB/C mice were randomly divided into 5 groups, with 20 mice in each group.The mice were inguinal injected with QGY-7701 cells, and then subcutaneous injected with Birinapant (concentrations ranging from 0, 1, 5, 25, 125 µg/kg) in each group after two days, once every other day.On 18th day since first Birinapant injection, 10 mice were killed in each group to weigh tumor tissue and survival time was recorded from the remaining 10 mice.The effects of Birinapant on the growth of the tumor and the survival time of tumor-bearing mice were observed. RESULTS: Compared with the negative control (NC) group, the proliferation activity of QGY-7701 was inhibited significantly after Birinapant treatment and the apoptosis levels were increased significantly (P<0.01).The cell mitochondrial membrane potential was decreased and the karyotype was changed (P<0.01).At the same time, the transcription and expression levels of genes cellular inhibitor of apoptosis protein 1(cIAP-1), cellular inhibitor of apoptosis protein 2(cIAP-2), ras, raf, mek and erk were significantly decreased (P<0.01), while the expression levels of caspase-3 and caspase-9 genes were up-regulated (P<0.01).Compared with the model group (MG), the growth of the tumor was inhibited significantly and the survival time of the tumor-bearing mice was prolonged after Birinapant treatment (P<0.01). CONCLUSIONS: Birinapant can inhibit the expression of cIAP-1, cIAP-2 and the proteins of Ras-Raf-MEK-ERK signal pathways, so as to activate the mitochondria mediated endogenous apoptosis pathway.Birinapant shows a certain inhibitory effect on liver cancer.