This study aims to construct C57/B6-L and A549 cell lines that stably overexpress circu-lar RNA LAMP3(circLAMP3),laying the foundation for further research on the biological func-tions of circLAMP3.Total RNA was extracted and reverse transcripted into cDNA from C57/B6-L and A549 cells to amplify the full-length sequence of circLAMP3.Then,the fragments of cir-cLAMP3 were ligated into pLC5-ciR vector to obtain pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 recombinant plasmids.The lentiviruses expressing circLAMP3 were packaged on tran-sient transfected HEK293T cells.C57/B6-L and A549 cells were infected with lentiviruses to gen-erate cell lines overexpressing circLAMP3 through puromycin screening.To verify the overexpres-sion efficiency of circLAMP3 of cell lines,we performed the fluorescence microscopy,PCR amplifi-cation,quantitative PCR(qPCR),and Sanger sequencing experiments.The results indicated that the overexpression plasmids of pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 were suc-cessfully constructed.Strong green fluorescence was observed under a fluorescence microscopy.C57/B6-L and A549 cell lines showed a significant increase in the expression of circLAMP3 by PCR and qPCR methods.Sanger sequencing results showed that the junction site of circLAMP3 was correct.This study successfully constructed C57/B6-L and A549 cell lines overexpressing circLAMP3,providing biomaterials for further exploration of the biological function of circLAMP3 in influenza virus replication.

Objective : To investigate the differences in gut microbiota and metabolites between urban and rural middle school students and explore their significance in gut homeostasis , so as to establish a healthy lifestyle and diet for children. Methods : Fecal samples were collected from middle school students in Hefei ( n = 14) and Jixi county ( n = 18 , Southern Anhui) , aged 13. 0 - 13. 5 years. Stool samples were sequenced by 16S ribosomal DNA (LC⁃MS) , followed by bioinformatic analysis. Results : Lachnoclostridium and Anaerostipes were dominant in the urban students that had been reported to be associated with colorectal cancer, atherosclerosis , depression and other disorders. In the village children , Ruminococcaceae UCG⁃002 , Barnesiella and Eubacterium dominated. An increased proportion of these microbes were related to metabolism of bile acids , short⁃chain fatty acids , lipid and carbohydrate decomposition , and play an important role in maintaining immune balance and physiological function. Additionally , significant differences in gut metabolites of the two groups were noted , mainly in arachidonic acid metabolism , platelet activation , serotonin metabolism , vitamin absorption , primary bile acid metabolism and other pathways. Conclusion Adolescent students of urban and mountainous areas differ in gut microbiota and metabo⁃ lites. Rural children have a healthy bacterial flora and metabolites in guts due to a reasonable lifestyle and diet in comparison with the city children.

Eukaryotic translation initiation factor 4B (eIF4B) plays an important role in mRNA translation initiation, cell survival and proliferation in vitro, but the in vivo function is poorly understood. In this study, via various experimental techniques such as hematoxylin-eosin (HE) staining, flow cytometry, Western blotting, and immunohistochemistry, we investigated the role of eIF4B in mouse embryo development using an eIF4B knockout (KO) mouse model and explored the mechanism. We found that the livers, but not lungs, brain, stomach, or pancreas, derived from eIF4B KO mouse embryos displayed severe pathological changes characterized by enhanced apoptosis and necrosis. Accordingly, high expression of cleaved-caspase 3, and excessive activation of mTOR signaling as evidenced by increased expression and phosphorylation of p70S6K and enhanced phosphorylation of 4EBP1, were observed in mouse embryonic fibroblasts and fetal livers from eIF4B KO mice. These results uncover a critical role of eIF4B in mouse embryo development and provide important insights into the biological functions of eIF4B in vivo.
Animals ; Apoptosis/genetics* ; Caspase 3 ; Eosine Yellowish-(YS) ; Eukaryotic Initiation Factors/metabolism* ; Fibroblasts ; Hematoxylin ; Liver/metabolism* ; Mice ; Ribosomal Protein S6 Kinases, 70-kDa/genetics* ; TOR Serine-Threonine Kinases

Objective:To explore the effect of 1-methyltryptophan (1-MT) on lipopolysaccharide (LPS)-induced permeability and apoptosis of human umbilical vein endothelial cells (HUVECs).Methods:HUVECs were treated with phosphate buffer saline (PBS, control group), 1 μg/mL LPS (LPS group), and LPS combined with 1 mmol/L 1-MT (1-MT group). The expression levels of the p120 concatemer (p120ctn), vascular endothelial (VE) cadherin, caspase-3, and DNA repair enzyme polyadenylate ribose polymerase-1 (PARP) after incubation at 8 h were detected using Western blot. The concentrations of kynurenine (Kyn) after incubation at 2, 4, 6, and 8 h were measured by high-performance liquid chromatography, and indoleamine2, 3-dioxygenase (IDO) activity was calculated. Comparisons among groups were performed using the LSD- t test. Results:Compared with the control group, the expression of caspase-3 [(74.01±7.91)% vs (157.14±7.63)%, P<0.01] and the concentration of Kyn were significantly up-regulated, while the expression of p120ctn [(49.12±2.15)% vs (37.61±1.80)%, P<0.01], VE-cadherin [(107.70±7.01)% vs (90.66±2.58)%, P=0.027], and PARP-1 [(67.95± 3.08)% vs (57.93±5.26)%, P=0.038] were significantly down-regulated, and IDO activity was significantly increased in the LPS group ( P<0.05). Compared with the LPS group, the expression of caspase-3 [(157.14±7.63)% vs (110.74±7.89)%, P<0.01] was significantly down-regulated, while the expression of p120ctn [(37.61±1.80)% vs (47.19±0.82)%, P<0.01], VE-cadherin [(90.66±2.58)% vs (107.27±9.89)%, P=0.029], and PARP-1 [(57.93±5.26)% vs (74.12±4.90)%, P=0.005] were significantly up-regulated, and the activity of IDO was significantly decreased over time in the 1-MT group ( P<0.05). No significant differences were observed between the PBS and 1-MT groups in the protein levels of p120ctn, VE-cadherin, and PARP-1 protein as well as Kyn concentration and IDO activity ( P>0.05), while the expression of caspase-3 was increased in 1-MT group ( P=0.001). Conclusions:LPS aggravates the permeability of HUVECs, which can be reversed by 1-MT via inhibiting IDO activity and reducing Kyn concentrations. Moreover, 1-MT can also reduce apoptosis, which may be via increasing the expression of PARP-1 and reducing the expression of caspase-3, thus protecting endothelial cells.

Long noncoding RNAs (lncRNAs) are a class of RNA molecules that are greater than 200 nt in length and do not have protein-coding capabilities or encode micropeptides only. LncRNAs are involved in the regulation of cell proliferation, differentiation, apoptosis and other biological processes, and are closely associated with the occurrence, recurrence and metastasis of a variety of malignant hematologic diseases. This article summarizes the function, regulatory mechanism and potential clinical application of lncRNAs in leukemia. In general, lncRNAs regulate the occurrence and development of leukemia and the multi-drug resistance in chemotherapy through epigenetic modification, ribosomal RNA transcription, competitive binding with miRNA, modulating glucose metabolic pathway, and activating tumor-related signaling pathway. Studies on lncRNAs provide new references for understanding the pathogenesis of leukemia, uncovering new prognostic markers and potential therapeutic targets, and addressing the problems of drug resistance and post-treatment recurrence in patients in clinical treatment of leukemia.
Cell Proliferation ; Humans ; Leukemia/genetics* ; MicroRNAs ; Neoplasms ; RNA, Long Noncoding/genetics*

Human acute leukemia (AL) is a clonal malignancy with abnormal hematopoietic stem cells. Clinically, AL is very difficult to cure due to its sudden onset and short course of disease progression. Previous studies have shown that eukaryotic initiation factor 4B (eIF4B) plays a critical role in the development of chronic leukemia. However, the involvement of eIF4B in human acute leukemia is still largely unknown. Therefore, we studied eIF4B function and its regulatory mechanism in human acute leukemia. We found that phosphorylation levels of eIF4B in acute leukemia cells were significantly reduced in response to treatment with either LY294002 (PI3K inhibitor), AKTi (AKT inhibitor) or SMI-4A (Pim inhibitor). Co-treatment with inhibitors targeting JAK/STAT5/Pim and PI3K/AKT/mTOR signaling dramatically promoted apoptosis of acute leukemia cells by downregulating eIF4B phosphorylation. Furthermore, in vitro and in vivo functional experiments showed that eIF4B played an important anti-apoptosis role in the acute leukemia cells by regulating the expression of anti-apoptotic proteins Bcl-2 and Bcl-XL. In contrast, silencing eIF4B inhibited the growth of acute leukemia cells as engrafted tumors in nude mice. Taken together, our results indicate the synergistic role of JAK/STAT5/Pim and PI3K/AKT/mTOR signaling pathways in regulating eIF4B phosphorylation in acute leukemia, and highlight eIF4B as a candidate therapeutic target for treatment of acute leukemia.
Animals ; Apoptosis ; Cell Line, Tumor ; Leukemia ; Mice ; Mice, Nude ; Phosphatidylinositol 3-Kinases/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; STAT5 Transcription Factor/metabolism*

Objective:To explore the psychological state and affected factors of elderly patients with hip fractures.Methods:A retrospective analysis of 156 elderly hip fracture patients(>65 years) admitted to the Department of Orthopaedics, the First Affiliated Hospital of Harbin Medical University from January 2016 to August 2019 was performed. General and psychological information were collected by questionnaire.General information included age, gender, education, whether surgery, length of stay.SCL-90, a self-assessment scale, was chosen as the psychological test to analyzed the elderly hip fracture patients′ psychological status during hospitalization and the norms of SCL-90 in Chinese which were established in 1986 were used as the control group. The prognostic factors were examined by univariate and multivariate analysis.Results:Somatization, interpersonal sensitivity, depression, anxiety, paranoid factor scores, and total scores of the elderly hip fracture patients were significantly higher than control group(all P=0.00).Univariate analysis and logistic regression analysis showed that non-surgery treatment and more than 10 days of hospitalization were independent prognostic factors that affected the psychological state of elderly hip fracture patients (all P=0.00). Conclusion:Elderly patients hospitalized with osteoporosis and hip fractures are prone to have negative emotional and psychological changes.The length of hospitalization and the choice of treatment can affect patients′ psychological state, suggesting that effective psychological intervention is necessary.

Objective:To explore the psychological state and affected factors of elderly patients with hip fractures.Methods:A retrospective analysis of 156 elderly hip fracture patients(>65 years) admitted to the Department of Orthopaedics, the First Affiliated Hospital of Harbin Medical University from January 2016 to August 2019 was performed. General and psychological information were collected by questionnaire.General information included age, gender, education, whether surgery, length of stay.SCL-90, a self-assessment scale, was chosen as the psychological test to analyzed the elderly hip fracture patients′ psychological status during hospitalization and the norms of SCL-90 in Chinese which were established in 1986 were used as the control group. The prognostic factors were examined by univariate and multivariate analysis.Results:Somatization, interpersonal sensitivity, depression, anxiety, paranoid factor scores, and total scores of the elderly hip fracture patients were significantly higher than control group(all P=0.00).Univariate analysis and logistic regression analysis showed that non-surgery treatment and more than 10 days of hospitalization were independent prognostic factors that affected the psychological state of elderly hip fracture patients (all P=0.00). Conclusion:Elderly patients hospitalized with osteoporosis and hip fractures are prone to have negative emotional and psychological changes.The length of hospitalization and the choice of treatment can affect patients′ psychological state, suggesting that effective psychological intervention is necessary.

At present, the application of the robot assisted surgery system in the surgical treatment of esophageal cancer is gradually emerging, and it is more and more widely used and recognized in the field of surgery. According to the domestic and foreign literatures, the robot has many advantages, and robotic assisted esophageal cancer surgery has been proved to be safe and effective, and its short-term efficacy is significantly better than thoracotomy. Other studies have shown that in long-term follow-up, the effect is comparable to video-assisted thoracoscopic surgery. In this paper, the author are systematically reviewed the development history of the robot assisted surgery system, the effect of robotic assisted esophagectomy on safety, surgical method, short-term efficacy and long-term prognosis. The traditional open surgery and thoracoscopic laparoscopic esophagectomy has been carried on the detailed comparison to provide some advice and theoretical basis for esophageal cancer surgery robot system.

Objective    To collect the data of esophageal foreign body patients, and to evaluate the clinical effects of two different surgical methods in our hospital. Methods    The clinical data of 294 patients who were treated in Gansu Provincal Hospital from January 2012 to June 2018 were analyzed retrospectively. The clinical data were collected and analyzed by SPSS 22.0. In order to to evaluate the efficacy of flexible esophagoscope (FE) and rigid esophagoscope (RE) in the treatment of esophageal foreign bodies.The patients were divided into two groups: a RE group including 118 patients with 62 males and 56 females at age of  6 (3-37) years and a FE group including 176 patients with 84 males and 92 females at age of 6 (3-59) years. Results    There was no significant difference in age, age stratification, gender and foreign body type between the two groups. There was a statistical difference in the initial clinical symptoms (P=0.041) or in esophageal foreign bodies position (P=0.037) between the two groups. The success rate of foreign body removal was similar between the two groups (P=0.632). The success rate was 88.9% (105/118) in the RE group, 87.5% (154/176) in the FE group. The operation time was significantly longer in the RE group than that in the FE group (10.8 ±17.4 min vs. 17.5±21.6 min, P<0.001). The postoperative hospitalization time in the RE groups was longer than that in the FE group (21.5 ±24.2 hours vs. 12.5 ±21.3 hours, P<0.05). There was a statistical difference in the incidence of postoperative complications between the two groups (P=0.034). In the RE group, the main complication was mucosal edema (15.3%). And the rate of bleeding was higher (15.9%) in the FE group. There were 30 patients (25.5%) in the RE group with minor postoperative complications versus the FE group with 40 patients (22.7%); and 1 patient (0.8%) in the RE group with severe complications versus the FE group with 5 paients (2.8%). Conclusion    Based on the analysis of this study, it is found that RE has higher safety. But the indications are strict, the professional requirements of the operator and the selection of patients are stronger. The FE is convenient to use, the operation crowd is wide, and the suitable crowd is wide. Therefore, for specific patients, after improving the relevant examination and preoperative evaluation of patients, clinicians need to choose appropriate surgical methods to ensure the success of the operation, and reduce the postoperative complications as far as possible.