Objective·To establish a mouse model with dormant cancer and no obvious metastasis by combining the preimmune strategy with the mVenus-p27K-cell G0 phase indicator system,the DTR-HSV/TK suicide gene system,and the Luc2-tdTomato tracer system.Methods·The KPC1199 mouse pancreatic cancer cell line was transfected with the mVenus-p27K-cell G0 phase indicator system,the DTR-HSV/TK suicide gene system,and the Luc2-tdTomato tracer system to construct a stable expression cell line,KPC1199-PDL.After being cultured in the serum-free condition,KPC1199-PDL cells were sorted into mVenus(+)cells and mVenus(-)cells by flow cytometry,and the expression of G0 phase-related genes was verified by real-time fluorescence quantitative PCR(qPCR).Sensitivity of KPC1199-PDL cells to diphtheria toxin(DTX)and ganciclovir(GCV)was evaluated by CCK-8 assay.A transsplenic portal vein-hepatic metastasis model was constructed in wild-type C57BL/6 mice to validate the function of KPC1199-PDL cells in vivo by immunofluorescence technology.The KPC1199-PDL cells were injected subcutaneously into C57BL/6 mice,followed by in situ injection of DTX and GCV to ablate subcutaneous tumors 5 d later,to obtain preimmunized mice.The transsplenic portal vein-hepatic metastasis models were constructed in these mice.Bioluminescence imaging was used to evaluate subcutaneous tumor ablation and hepatic metastasis in the mice,and immunofluorescence assay was used to detect the distribution and dormant state of tumor cells in the livers of preimmunize mice.Results·The three tool systems were stably expressed in KPC1199-PDL cells,and their proliferative ability was not affected.In the serum starving condition,some KPC1199-PDL cells expressed the mVenus protein,indicating entry into the G0 phase;the mVenus(+)cells sorted out by flow cytometry exhibited significantly higher expression of G0 phase-related genes(all P<0.05)and significantly lower expression of the proliferation-related gene compared with mVenus(-)cells(P<0.05).The CCK-8 assay demonstrated high sensitivity of KPC1199-PDL cells to DTX and GCV.In vivo experiments confirmed that KPC1199-PDL cells could be effectively traced through tdTomato protein expression,and could indicate entry into the G0 phase through mVenus protein expression.Following subcutaneous tumor implantation and drug ablation,preimmunized mice were successfully obtained.In the subsequent transsplenic portal vein-hepatic metastasis model,no metastatic signals were observed in the liver by bioluminescence imaging,but single or small clusters of G0 phase tumor cells expressing both mVenus and tdTomato,not expressing the proliferation marker Ki67,were detected in liver tissue sections by immunofluorescence analysis.Conclusions·A recognizable and traceable dormant cancer model is constructed with the combination of the preimmune mouse model of pancreatic cancer,the mVeneus-p27K-indicator system,the DTR-HSV/TK suicide gene system,and the Luc2-tdTomato tracer system.

Humans ; Esophageal Squamous Cell Carcinoma/pathology* ; Esophageal Neoplasms/pathology* ; Carcinoma, Squamous Cell/pathology* ; Precancerous Conditions/pathology* ; Early Diagnosis

Objective:To investigate the impacts of conscious sedation and general anesthesia on the functional outcome after endovascular therapy (EVT) in elderly patients with acute ischemic stroke with large vessel occlusion (AIS-LVO).Methods:The clinical and imaging data of elderly patients with AIS-LVO (≥80 years) underwent EVT at the Affiliated Hospital of Nantong University from January 2020 to January 2023 were collected retrospectively. They were divided into conscious sedation group and general anesthesia group according to anesthesia modality, and divided into good outcome group (0-2 points) and poor outcome group (>2 points) based on the modified Rankin Scale score at 90 d after onset. The multivariate logistic regression analysis was used to investigate the impact of anesthesia modality on functional outcome after EVT. Results:A total of 77 elderly patients with AIS-LVO were enrolled, including 35 males (45.5%) and 42 females (54.5%); median age of 82.0 years (interquartile range, 80.0 to 84.0 years); the median baseline NIHSS score was 16.0 (interquartile range, 10.0-20.0). Conscious sedation was used in 21 cases (27.3%) and general anesthesia was used in 56 cases (72.7%); 17 (22.1%) had good outcome, while 60 (77.9%) had poor outcome. Compared with the general anesthesia group, the conscious sedation group had a longer procedure time (110.0 min vs. 89.0 min; P=0.049), but a higher rate of good outcome at 90 d (38.1% vs. 16.1%; P=0.038), a lower incidence of stroke-associated pneumonia (33.3% vs. 58.9%; P=0.045), and a lower proportion of patients who underwent tracheostomy after procedure (4.8% vs. 25.0%; P=0.046). Compared with the poor outcome group, the good outcome group had shorter procedure time (75 min vs. 99 min; P=0.033), lower incidence of stroke-associated pneumonia (29.4% vs. 58.3%; P=0.035), lower tracheotomy rate (0% vs. 25%; P=0.022), and a lower proportion of patients who received conscious sedation (47.1% vs. 21.7%; P=0.038). Multivariate logistic regression analysis showed that conscious sedation was an independent predictor of good outcome (odds ratio 0.090, 95% confidence interval 0.010-0.771; P=0.028). Conclusion:Conscious sedation may be more appropriate for elderly patients with anterior circulation AIS-LVO undergoing endovascular treatment.

Objective To investigate the effect and mechanism of miR-27a-3p on nerve cell apoptosis induced by oxygen glucose deprivation/reoxygenation(OGD/R)through regulation of Rho family GTPase 3(Rnd3)expression.Methods PC12 neurons were cultured in vitro and reoxygenated for 3,6,9 h and 12 h after 2 h oxygen glucose deprivation.Cell viability,miR-27a-3p expression and Rnd3 mRNA expression were assessed at each time point and the optimal reoxygenation time point was screened.After transfection of miR-27a-3p Mimic,miR-27a-3p Inhibitor and their negative control,transfection of shRnd3 and its negative control,or co-transfection of shRnd3 and miR-27a-3p Inhibitor through lentivirus,CCK-8 assay was used to detect cell activity.The apoptosis rate of the cells was detected using flow cytometry.Expression of miR-27a-3p and Rnd3 mRNA was detected by RT-qPCR.Expression of apoptosis-related protein and Rnd3 protein was detected by Western blot.The dual luciferase reporter assay confirmed the targeting relationship between miR-27a-3p and Rnd3.Results Upregulation of miR-27a-3p increased cell viability,decreased total cell apoptosis rate,suppressed pro-apoptotic proteins Cleaved Caspase-3(C-caspase-3)and Bax,and promoted expression of anti-apoptotic protein Bcl-2(P<0.05);The opposite result was found when down-regulating miR-27a-3p.The double luciferase reporter gene assay showed that Rnd3 was the target gene of miR-27a-3p.Down-regulation of Rnd3 increased cell viability,decreased the total rate of apoptosis,suppressed the pro-apoptotic protein C-caspase-3,Bax,and promoted expression of the anti-apoptotic protein Bcl-2(P<0.05).However,miR-27a-3p Inhibitor reversed the protective effect of shRnd3.Conclusion miR-27a-3p alleviates OGD/R-induced damage to PC12 neurons by targeting Rnd3 to inhibit cell apoptosis.

Nanotechnology has emerged as an ideal approach for achieving the efficient chemo agent delivery. However, the potential toxicity and unclear internal metabolism of most nano-carriers was still a major obstacle for the clinical application. Herein, a novel "core‒shell" co-assembly carrier-free nanosystem was constructed based on natural sources of ursolic acid (UA) and polyphenol (EGCG) with the EpCAM-aptamer modification for hepatocellular carcinoma (HCC) synergistic treatment. As the nature products derived from food-plant, UA and EGCG had good anticancer activities and low toxicity. With the simple and "green" method, the nanodrugs had the advantages of good stability, pH-responsive and strong penetration of tumor tissues, which was expected to increase tumor cellular uptake, long circulation and effectively avoid the potential defects of traditional carriers. The nanocomplex exhibited the low cytotoxicity in the normal cells

Neoadjuvant chemotherapy (NACT) is a vital part of the systemic treatment to breast cancer. With the formation of consensuses on NACT, controversial perspectives on NACT have been widely discussed, especially in the fields of indication and therapeutic strategy. To define the indication of NACT, blind obedience to the results of clinical trials is not recommended. Instead, indications of NACT should be strictly controlled based on the targets of the clinical practice. Oriented by the early effectiveness of NACT, various chemotherapy or local therapeutics for different molecular subtypes of breast cancer should be conducted to the patients with unsatisfied effect. What′s more, the evolvement of precision medicine accelerates the research of drugs and helps to form an individualized NACT plan. After clarifying the controversial opinions towards NACT in breast cancer, controlling the indication and optimizing the therapeutic strategy will improve the survival of breast cancer patients.

Neoadjuvant chemotherapy (NACT) is a vital part of the systemic treatment to breast cancer. With the formation of consensuses on NACT, controversial perspectives on NACT have been widely discussed, especially in the fields of indication and therapeutic strategy. To define the indication of NACT, blind obedience to the results of clinical trials is not recommended. Instead, indications of NACT should be strictly controlled based on the targets of the clinical practice. Oriented by the early effectiveness of NACT, various chemotherapy or local therapeutics for different molecular subtypes of breast cancer should be conducted to the patients with unsatisfied effect. What′s more, the evolvement of precision medicine accelerates the research of drugs and helps to form an individualized NACT plan. After clarifying the controversial opinions towards NACT in breast cancer, controlling the indication and optimizing the therapeutic strategy will improve the survival of breast cancer patients.

The prevalence of obesity-associated conditions raises new challenges in clinical medication. Although altered expression of drug-metabolizing enzymes (DMEs) has been shown in obesity, the impacts of obese levels (overweight, obesity, and severe obesity) on the expression of DMEs have not been elucidated. Especially, limited information is available on whether parental obese levels affect ontogenic expression of DMEs in children. Here, a high-fat diet (HFD) and three feeding durations were used to mimic different obese levels in C57BL/6 mice. The hepatic expression of five nuclear receptors (NRs) and nine DMEs was examined. In general, a trend of induced expression of NRs and DMEs (except for and ) was observed in HFD groups compared to low-fat diet (LFD) groups. Differential effects of HFD on the hepatic expression of DMEs were found in adult mice at different obese levels. Family-based dietary style of an HFD altered the ontogenic expression of DMEs in the offspring older than 15 days. Furthermore, obese levels of parental mice affected the hepatic expression of DMEs in offspring. Overall, the results indicate that obese levels affected expression of the DMEs in adult individuals and that of their children. Drug dosage might need to be optimized based on the obese levels.

Objective:To evaluate the effects of exogenous biliverdin (BV) on the expression of Litaf in PC12 cells subjected to oxygen-glucose deprivation and restoration (OGD/R).Methods:PC12 cells were seeded in a 96-well cell culture plate at a density of 1×10 4 cells/well for 3 days and were divided into 3 groups ( n=18 each) by a random number table method: control group (group C), OGD/R group, and biliverdin group (BV group). Group C was incubated in a 37 ℃ incubator (95% air+ 5%CO 2) for 6 h. To establish the OGD/R model, cells were incubated with sugar-free medium in a 37 ℃ incubator (95% air+ 5%CO 2) for 2 h, and the medium was then replaced with normal medium and cells were continuously incubated in a 37 ℃ incubator (95% N 2+ 5% CO 2). In BV group, 2 μg/ml biliverdin was added immediately after oxygen-glucose restoration.Cells in 6 wells in each group were selected at 6 h of restoration for determination of the expression of Litaf protein and mRNA (by real-time polymerase chain reaction) and tumor necrosis factor-alpha (TNF-α) concentration (by enzyme-linked immunosorbent assay). Results:Compared with group C, the expression of Litaf protein and mRNA was significantly up-regulated, and TNF-α concentration in supernatant was increased in group OGD/R ( P<0.05). Compared with group OGD/R, the expression of Litaf protein and mRNA was significantly down-regulated, and TNF-α concentration in supernatant was decreased in group BV ( P<0.05). Conclusion:The mechanism by which exogenous biliverdin reduces OGD/R damage to PC12 cells is related to inhibiting up-regulated expression of Litaf and alleviating the inflammatory responses.

Objective:To evaluate the effect of propofol anesthesia on autophagy in hippocampal neurons of newborn rats.Methods:Thirty-nine healthy Sprague-Dawley rats, aged 7 days, weighing 10-12 g, were divided into 3 groups ( n=13 each) using a random number table method: control group (group C), fat emulsion group (group F) and propofol group (group P). Normal saline 8 ml/kg was intraperitoneally injected for 5 consecutive days in group C. Medium-/long-chain fatty emulsion injection 8 ml/kg was intraperitoneally injected for 5 consecutive days in group F. Medium-/long-chain propofol injection 80 mg/kg was intraperitoneally injected for 5 consecutive days in group P. Five rats were sacrificed on 1st day after the end of propofol anesthesia, and hippocampal tissues were taken for determination of the expression of microtubule-associated protein 1 light chain 3B (LC3B) and Beclin-1 (by Western blot). The remaining rats in each group underwent the Morris water maze test on 19th day after the end of propofol anesthesia (30 days after birth), and the escape latency, percentage of time of staying at the target quadrant and the number of crossing the original platform were recorded. Results:Compared with group C, no significant change was found in the expression of hippocampal LC3B and Beclin-1, escape latency, percentage of time of staying at the target quadrant, and the number of crossing the original platform in group F ( P>0.05), and the expression of hippocampal LC3B and Beclin-1 was significantly up-regulated, the escape latency was prolonged, percentage of time of staying at the target quadrant was decreased, and the number of crossing the original platform was decreased in group P ( P<0.05 or 0.01). Conclusion:The mechanism by which propofol anesthesia causes long-term cognitive dysfunction may be related to promoting autophagy in hippocampal neurons of newborn rats.