OBJECTIVE:To establish a method for determining plasma concentrations of tanshinone ⅡA and use it for pharma-cokinetics study of tanshinone ⅡA in Beagle dogs. METHODS:HPLC was performed on the column of Phenomenex Luna C18, with the mobile phase of water(5 mmol/L of ammonium acetate+0.05% H3PO4)-acetonitrile at flow rate of 1.0 ml/min;the detec-tion wavelength was 270 nm,column temperature was 40 ℃ and the volume was 20 μl. 9 Beagle dogs were randomly divided into tanshinone ⅡA low,medium and high dose groups(2,4 and 8 mg/kg),blood was respectively taken from forelimb venous blood to determine the plasma concentration before and after 2,5,10,15,20,30,45,60,75,90 and 120 min of iv administration. DAS 2.0 software was used to calculate the pharmacokinetic parameters. RESULTS:The linear range of tanshinoneⅡA was 0.097 5-12.50 μg/ml (r=0.999 8);the RSDs of precision and stability tests were all less than 10%;the method recovery was 100.4%-107.2%,and extraction recovery was 90.2%-92.3%. The t1/2αin tanshinoneⅡA low,medium and high dose groups were respective-ly(1.01±0.27),(1.03±0.46)and(1.51±0.65)min;t1/2β were respectively(16.25±4.78),(22.42±9.32)and(24.45±12.02) min;AUC0-120 min were respectively(150.88±45.25),(305.44±92.55)and(643.67±178.27)μg·min/ml;and CL were respective-ly(12.01±4.36),(12.78±5.06)and(12.17±5.41)ml/(min·kg). CONCLUSIONS:The precision,stability and recovery of the method are all in line with the determination requirements of biological samples,and tanshinone ⅡA showed a good linear relation-ship with dose in Beagle dogs in AUC0-120 min.

In this study, we evaluated the development potential of caprine mammary gland epithelial cells (CMGECs) after transfection and nuclear transfer into enucleated, ovulated oocytes. We first isolated CMGECs from udders of lactating goats which were transfected with expression plasmid for human lacterrin and selected by G418. Then we chose sixteen neomycin resistant lines and induced them with prolactin for the expression of human lactoferrin checked by Western blotting. The donor cells, expressing human lactoferrin of 75 kD, were fused and activated with enucleated ovulated oocytes. Pronuclear-stage reconstructed embryos were transferred into the oviducts of 16 recipient goats. There were fourteen (87.5%), thirteen (81.3%), and ten (62.5%) pregnancies confirmed pregnant by ultrasound on Day 30, 60, and 90, respectively. Three recipients carried the pregnancies to term and delivered one goat each. Nested PCR-RFLP analysis confirmed that all of the kids were clones of the donor cells. These results demonstrated that CMGECs after transfection remain totipotent for nuclear transfer.
Animals ; Cloning, Organism ; methods ; Epithelial Cells ; cytology ; Female ; Goats ; Humans ; Lactoferrin ; biosynthesis ; Mammary Glands, Animal ; cytology ; Nuclear Transfer Techniques ; Pregnancy ; Transfection

Ferroptosis is a form of programmed cell death driven by iron-dependent lipid peroxidation and is closely associated with a wide range of biological processes, such as aging, immunity, and cancer. Tumor multidrug resistance, especially resistance to apoptosis, has prompted the urgent search for a new antitumor treatment option. Remarkable progress has been made in the study of the role of ferroptosis in antitumor, especially the interaction with immune cells. Studying immunotherapy based on ferroptosis pathways has become a new direction in antitumor research. In this work, we review the role of ferroptosis in immune cells and antitumor immunotherapy to provide new ideas for ferroptosis-mediated antitumor immunotherapy.

Objective:To establish a mortality risk prediction model of severe bacterial infection in children and compare it with the pediatric early warning score (PEWS), pediatric critical illness score (PCIS) and pediatric risk of mortality score Ⅲ (PRISM Ⅲ).Methods:A total of 178 critically ill children were selected from the PICU of the Children's Hospital of Nanjing Medical University from May 2017 to June 2022. After obtaining the informed consent of the parents/guardians, basic information such as sex, age, height and weight, as well as indicators such as heart rate, systolic blood pressure and respiratory rate were collected from all children. A standard questionnaire was used to score the child 24 h after admission to the PICU. The children were divided into the survival and death groups according to their survival status at 28 d after admission. A mortality risk prediction model was constructed and nomogram was drawn. The value of the mortality risk prediction model, PEWS, PCIS and PRISM in predicting the risk of death was assessed and compared using the receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC).Results:Among the 178 critically ill children, 11 cases were excluded due to severe data deficiencies and hospitalization not exceeding 24 h. A total of 167 children were included in the analysis, including 134 in the survival group and 33 in the death group. A mortality risk prediction model for children with severe bacterial infection was constructed using pupillary changes, state of consciousness, skin color, mechanical ventilation, total cholesterol and prothrombin time. ROC curve analysis showed that the AUCs of mortality risk prediction model was 0.888 ( P<0.05). The AUCs of PEWS, PCIS and PRISM Ⅲ in predicting death in children with severe bacterial infection were 0.769 ( P< 0.05), 0.575 ( P< 0.05) and 0.759 ( P< 0.05), respectively. Hosmer-Lemeshow goodness-of-fit test showed the best agreement between risk of death and PEWS predicted morbidity and mortality and actual morbidity and mortality (χ 2 = 5.180, P = 0.738; χ 2 = 4.939, P = 0.764), and the PCIS and PRISM Ⅲ predicted mortality rates fitted reasonably well with actual mortality rates (χ 2= 9.110, P= 0333; χ 2 = 8.943, P= 0.347). Conclusions:The mortality risk prediction model for predicting the death risk has better prognostic value than PEWS, PCIS and PRISM Ⅲ for children with severe bacterial infection.

Objective To evaluate the efficacy and further analyze the application prospects of the combined multitarget fecal FIT-DNA assay in the early screening of colorectal cancer. Methods Subjects were selected from a population attending the Inner Mongolia Medical University Hospital. Each subject underwent a combined multi-target fecal FIT-DNA test (experimental group), a serum tumor marker test and enteroscopy (control group). The pathological results were used as the gold standard to evaluate the efficacy of novel fecal molecular testing techniques for colorectal cancer screening with timely intervention given to screen positive individuals. Results The data of 115 individuals were analyzed. Serum tumor markers test had a sensitivity of 63.2% (43/68) and a specificity of 74.5% (35/47). The enteroscopy had a sensitivity of 97.1% (66/68) and a specificity of 80.7% (38/47); the combined multitarget fecal FIT-DNA test had a sensitivity of 89.7% (61/68) and a specificity of 87.2% (41/47). Conclusion The sensitivity and specificity of multitarget fecal FIT-DNA combined detection are better than those of serum tumor marker detection. Although its sensitivity is lower than enteroscopy, its operation is simpler and can be tested at home.