Objective To explore the feasibility of long non-coding RNAs (lncRNAs) as biomarkers for radiation-induced intestinal injury. Methods Mice were exposed to 15 Gy of ⁶⁰Co γ-rays to the abdominal area. The pathological changes in intestinal tissues were analyzed at 72 h post-irradiation to confirm the successful establishment of the radiation-induced intestinal injury model. Real-time quantitative PCR was conducted to detect the expression of candidate radiation-responsive lncRNAs in the jejunum, jejunal crypts, colon tissues, and plasma of irradiated mice. Human intestinal epithelial cell line HIEC-6 and human colon epithelial cell line NCM460 were exposed to 0, 5, 10, and 15 Gy of ⁶⁰Co γ-rays. The expression levels of candidate lncRNAs were measured at 4, 24, 48, and 72 h post-irradiation to observe their changes with the irradiation dose. Results Pathological analysis showed that abdominal irradiation with 15 Gy successfully established an acute radiation-induced intestinal injury mouse model. Real-time quantitative PCR showed that Dino, Lncpint, Meg3, Dnm3os, Trp53cor1, Pvt1, and Neat1 were significantly upregulated following the occurrence of radiation-induced intestinal injury (P < 0.05). Among them, Meg3 and Dnm3os in mouse plasma were significantly upregulated (P < 0.05), while Gas5 was significantly downregulated (P < 0.05). In HIEC-6 and NCM460 cells, the expression levels of DINO, MEG3, DNM3OS, and GAS5 showed dose-dependent patterns at certain time points (P < 0.05). Conclusion The lncRNAs encoded by MEG3, DNM3OS, and GAS5 in intestinal epithelial cells are responsive to ionizing radiation. Consistent differential expression changes were detected in mouse plasma and intestinal tissues, indicating their potential as biomarkers for radiation-induced intestinal injury.

Objective To develop a highly sensitive and rapid nucleic acid detection method for the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2). Methods We designed,developed,and manufactured an integrated disposable device for SARS-CoV-2 nucleic acid extraction and detection.The precision of the liquid transfer and temperature control was tested.A comparison between our device and a commercial kit for SARS-Cov-2 nucleic acid extraction was performed using real-time fluorescence reverse transcription polymerase chain reaction(RT-PCR).The entire process,from SARS-CoV-2 nucleic acid extraction to amplification,was evaluated. Results The precision of the syringe transfer volume was 19.2±1.9 μL(set value was 20),32.2±1.6(set value was 30),and 57.2±3.5(set value was 60).Temperature control in the amplification tube was measured at 60.0±0.0 ℃(set value was 60)and 95.1±0.2 ℃(set value was 95)respectively.SARS-Cov-2 nucleic acid extraction yield through the device was 7.10×106 copies/mL,while a commercial kit yielded 2.98×106 copies/mL.The mean time to complete the entire assay,from SARS-CoV-2 nucleic acid extraction to amplification detection,was 36 min and 45 s.The detection limit for SARS-CoV-2 nucleic acid was 250 copies/mL. Conclusion The integrated disposable devices may be used for SARS-CoV-2 Point-of-Care test(POCT).

OBJECTIVE To explore the effects of SMAD2/3 expression on radiotherapy and chemotherapy sensitivity in nasopharyngeal carcinoma(NPC)cells.METHODS The expression of SMAD2/3 was detected by immunofluorescence staining in the tissues of NPC,Western blot and qRT-PCR were used to detect the expression of SMAD2/3 in four human NPC cell lines(CNE-2,5-8F,Hone1,HK-1)and two human normal nasopharyngeal epithelial cells(HNEpC,NP69).The CNE-2 cell line(shSMAD2/3)and its control cell line(shSMAD2/3-NC)were constructed by lentiviral transfection.The transfected cells were treated with 4 Gy irradiation,10 μmol/L cisplatin chemotherapy and 4 Gy irradiation plus 10 μmol/L cisplatin chemotherapy respectively.The cell proliferation activity was detected by colony formation assay,the ability of cell migration was detected by scratch test,the ability of cell invasion was detected by Transwell test,and the phosphorylation level of PI3K/AKT/mTOR pathway was detected by Western blot.Then,PI3K agonist 740 Y-P was added to the experiment.RESULTS SMA2D/3 were highly expressed in NPC tissues and cell lines(all P<0.05).Compared with shSMAD2/3-NC cells,shSMAD2/3 cells were more sensitive to radiotherapy and chemotherapy,and their proliferation,migration and invasion were significantly inhibited(all P<0.05).The phosphorylation of PI3K/AKT/mTOR pathway was significantly decreased in shSMAD2/3 cells(all P<0.05).740 Y-P significantly increased the phosphorylation levels of the PI3K/AKT/mTOR pathway in the shSMAD2/3 group cells,effectively reversing the inhibition of cell viability by SMAD2/3 knockdown,and reducing the sensitivity of cells to radiotherapy and chemotherapy(all P<0.05).CONCLUSION SMAD2/3 is highly expressed in nasopharyngeal carcinoma,and down-regulation of SMAD2/3 expression increases the of nasopharyngeal carcinoma cell lines to radiotherapy and chemotherapy by reducing the phosphorylation levels of the PI3K/AKT/mTOR pathway.

To establish a recombinase aided amplification ( RAA) assay for Plasmodium, we use the RAA assay to screen Plasmodium. Universal primers and probe were designed based on the specific fragment of the 18S rDNA gene of Plasmodium. We screened the primers and detected the specificity of the method. We screened four better amplification primers. RAA was performed at 37℃ in a short time (40 minutes) with good specificity. The RAA assay for Plasmodium detection appears to be applicable in ports.

BACKGROUND: T cells are believed to play an important role in anti-infection, anti-tumor and immune function. However, the mechanism underlying the differentiation and development remains poorly understood. OBJECTIVE: To investigate the distribution of T cells in nude mice that are jointly transplanted human thymus and cord blood and the reconstruction of the immune function. METHODS: Thirty Balb/c nu/nu nude mice were randomly divided into two groups: an experimental group and a control group. In the experimental group, human thymus tissue was transplanted into the renal capsule of nude mice. Two weeks later, freshly isolated human cord blood CD34+ cells suspension was back perfused into the nude mice via the vein. In the control group, CD34+ cells transplantation was performed directly without thymus transplantation. After 60 days of breeding, the immune function of nude mice was detected in two groups. RESULTS AND CONCLUSION: Human thymus tissue in the renal capsule of nude mice survived and expressed CD3 and HLA-DR molecule. In the experimental group, CD3+ cells which distributed in the form of dots were observed in the mouse spleen. The proportion of CD3+, CD4+, CD8+, and CD4+CD25+ cells were significantly higher in the experimental group than in the control group. The nude mice from the experimental group rejected human gastric cancer BGC823 cells, while those from the control group did not. These findings demonstrated that combined human thymus and CD34+ cell transplantation allow nude mice to acquire T cell-mediated cellular immune function and possess the ability of anti-tumor.

OBJECTIVETo detect expression of p-glycoprotein (P-gp), multidrug resistance-associated protein (MRP), and glutathione S-transferase (GST-pi), and to evaluate its clinical significance in neuroblastoma (NB).

METHODSSP immunohistochemical technique was used to investigate expression of P-gp, MRP, and GST-pi in 70 cases of NB.

RESULTSThe frequency of expression of P-gp, MRP, and GST-pi was 61.4%, 38.6%, and 51.4%, respectively. The coexpression rate of P-gp and MRP, P-gp and GST-pi, MRP and GST-pi, P-gp, MRP and GST-pi was 32.9%, 35.7%, 27.1%, and 24.3%, respectively. Significant positive correlation was observed between P-gp and MRP expression (P = 0.001), and between MRP and GST-pi expression (P = 0.012), but no correlation was found between P-gp and GST-pi expression. The expression of P-gp and MRP was higher in tumors from patients over 1 year old compared with those less than 1 year old at diagnosis (P = 0.01, 0.018, respectively). MRP expression was higher in tumors from the metastatic than the non metastatic groups (P = 0.015). All tested proteins showed significant relationship to the differentiation of the tumor (P = 0.006, 0.000, 0.019, respectively), but no correlation was found to the stage of NB or sex of the patients. MRP expression was significantly related to the reduction of both median survival time and the two-year cumulative survival (P = 0.02). In contrast, P-gp and GST-pi expression had no correlation with survival.

CONCLUSIONSThe intrinsic multidrug resistance of NB involves the combined effects of P-gp, MRP, and GST-pi. MRP expression may be an important parameter in predicting the prognosis of patients with NB.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; Adolescent ; Child ; Child, Preschool ; Female ; Gene Expression ; Glutathione S-Transferase pi ; Glutathione Transferase ; biosynthesis ; Humans ; Immunohistochemistry ; Infant ; Isoenzymes ; biosynthesis ; Male ; Multidrug Resistance-Associated Proteins ; biosynthesis ; Neuroblastoma ; metabolism ; mortality ; Survival Rate