Objective To examine the genome-wide identification of PU. 1 binding sites and target genes before and after hemin-induced differentiation in K562 cells. Methods K562 cells were treated with hemin for 72 h to induce cell differentiation. High-throughput sequencing was performed following chromatin immunoprecipitation with PU. 1 antibody in untreated and hemin-treated cell groups. The binding sites of PU. 1 in K562 cells before and after differentiation were analyzed by bioinformatics. Gene annotation, gene function analysis, KEGG pathway analysis and target genes prediction were performed. Results The results showed that there were 3 107 and 1 897 binding sites of PU. 1 before and after differentiation respectively. Among them, 843 binding sites were identified in both untreated and differentiated K562 cells. KEGG pathway analysis showed that the NF-κB signaling pathway, growth hormone synthesis, secretion and transport pathway, and MAPK signaling pathway were the unique signaling pathways related to PU. 1 binding after differentiation. Totally, 1 210 PU. 1 binding sites changed during differentiation, corresponding to 179 target genes. Moreover, the PU. 1 target genes related to differentiation included globin genes, autophagy-related genes, microRNA and non-coding RNA, and transcription factors. Motif analysis showed that PU. 1 tended to bind a specific ACTTCC sequence. Conclusion The genomic binding sites of PU. 1 changed significantly after hemin-induced differentiation in K562 cells. Our study provided a basis for further research on the function of PU. 1 in hematopoietic cell

Objective To study the protein degradation mechanism of the small nucleolar ribonucleoprotein RRP9. Methods Tissue expression profiling was used to analyze the species-specific and tissue-specific expression of RRP9 in normal tissues of mice and humans, and the differential expression of RRP9 in normal colorectal tissues and colorectal cancer tissues. Cycloheximide (CHX), a protein synthesis inhibitor, was used to detect the half-life of RRP9 in colorectal cancer cells. After inhibiting the autophagy-lysosome pathway, the expression of RRP9 was detected to understand the effect of autophagy-lysosomal pathway on RRP9. Finally, immunofluorescence and co-immunoprecipitation were used to detect the interaction between LC3b and RRP9. Results RRP9 is a long-lived protein. Inhibition of autophagy-lysosomal pathway increased the level of RRP9. There was an interaction between RRP9 protein and LC3b in colorectal cancer cells. Conclusion The autophagy-lysosomal pathway regulates the degradation of RRP9 protein.

Objective To explore the effects of long non-coding RNA (lncRNA) -H19 on the proliferation, invasion and cell cycle of thyroid cancer cells. Methods The expression of lncRNA-H19 in cancer tissues, para-cancerous tissues, normal human thyroid epithelial cells (TEC) and thyroid cancer cells (KAT18, FTC133, BCPAP, TPC-1) were detected. The effects of silencing lncRNA-H19 on cell proliferation, invasion and cell cycle were detected. Results The lncRNA-H19 was highly expressed in thyroid cancer tissues and cell lines. The expression levels of lncRNA-H19 among thyroid cancer patients with different tumor sizes, N staging and TNM staging were significantly different (P< 0. 05). After silencing lncRNA-H19, the cell proliferation rate, clone formation rate, number of migrating cells and number of microvessel formation at 48 h and 72 h were significantly decreased (P< 0. 05), the expression levels of Ki67, PCNA, N-cadherin, Fibronectin, VEGF, MMP-9, MMP-14, cyclin A, CDK2, CDK4 and cyclin D1 proteins were significantly decreased (P< 0. 05), and the expression level of E-cadherin protein was significantly increased (P< 0. 05). Conclusion Silencing lncRNA-H19 could regulate the proliferation, invasion and cell cycle of FTC133 by regulating the expression of cell cycle related proteins.

Objective To explore the possible mechanisms by which lidocaine inhibits the activity and cisplatin resistance of human breast cancer cells by down-regulating ATP binding cassette transporter G family member 2 (ABCG2) in vitro. Methods The effects of lidocaine on cell proliferation and cisplatin intervention were verified in MDA-MB-231 and MCF-7 cells by CCK-8, EdU and TUNEL staining. The effects of lidocaine on the expressions of ABCG2, apoptosis proteins and drug resistance proteins were detected by RT-PCR and Western blotting; the phosphorylation of PI3K/ Akt proteins was detected by Western blotting. The effects of ABCG2 overexpression on the proliferation and apoptosis of breast cancer cells after intervention with lidocaine and cisplatin were verified by CCK-8 and TUNEL. Results After intervention with 0. 5 mmol / L lidocaine, the survival rate of breast cancer cells and the number of EdU-positive cells were significantly decreased (P< 0. 05). The survival rate of cells and the Bcl-2 expression were significantly decreased, the apoptosis rate and the expression levels of cleaved-PARP, cleaved-caspase-3 and Bax were significantly increased in lidocaine group and cisplatin group when compared with the control group. These changes were more obvious in cisplatin + lidocaine group (P < 0. 05). After lidocaine treatment, the expression levels of ABCG2 mRNA and protein in cancer cells were decreased (P< 0. 05). The expression levels of ABCG2, P-gp, MRP1 and MRP2 proteins were significantly decreased in cisplatin + lidocaine group when compared with the control group (P< 0. 05). The survival rate of cells and the levels of p-PI3K/ PI3K and p-AKT / AKT were significantly decreased (P < 0. 05), while the apoptosis rate was significantly increased in lidocaine group as compared to cisplatin group (P< 0. 05). The survival rate of cells and the levels of p-PI3K/ PI3K and p-AKT / AKT were significantly increased (P < 0. 05 ), while the apoptosis rate was significantly decreased in ABCG2 overexpressed group compared to the empty plasmid group (P< 0. 05). Conclusion Lidocaine could inhibit the proliferation of breast cancer cells and increase their sensitivity to cisplatin by inhibiting ABCG2 expression and activation of PI3K/ AKT signaling pathways.

Objective To investigate the expression of c-Ski and its role in p38 regulation in the endothelial-to-mesenchymal transition ( EndMT) in human coronary artery endothelial cells, which may provide novel targets for prevention and treatment of myocardial fibrosis. Methods The EndMT model was constructed in HCAECs using TGF-β1. The expression of c-Ski and EndMT markers was detected by quantitative real-time PCR and Western blotting. Results c-Ski was down-regulated after TGF-β1 treatment in the HCAECs. Overexpression of c-Ski suppressed TGF-β1-induced EndMT in HCAECs. The overexpression of c-Ski down-regulated the expression of α-SMA and vimentin, and up-regulated the expression of CD31 in TGF-β1-induced HCAECs ( P < 0. 01 ). The treatment with p38 inhibitor SB203580 further promoted TGF-β1-induced-EndMT in HCAECs; p38 signaling was activated after overexpression of c-Ski. Use of p38 inhibitor SB203580 could partially rescue the inhibitory effect of c-Ski overexpression on TGF-β1-induced EndMT in HCAECs. Conclusion c-Ski suppressed the TGF-β1-induced EndMT of HCAECs via regulating p38 activation. 

Objective To identify the precise ABO blood type subgroup with gene sequencing and discuss the relationship between the serology and genotype in CisAB / B blood subgroup. Methods We screened ABO blood type of blood donors in the blood center. Samples were first examined with a routine serological method. Those with ambiguous results were further examined with genotyping to identify ABO subgroups. The exon 6, exon 7 and the flanking region of ABO gene as well as its promoter and enhancer region were sequenced by PCR. Mutations found in the PCR were further confirmed by haplotype sequencing method. Results Genotype of the two cases were both CisAB01 / B101. Serological test showed the two cases to be A2B and AxB subgroup. Sequences of the promoter region and enhancer region of the two cases were the same. Conclusion It is difficult to identify CisAB only by serological method. DNA sequencing combined with serological testing is able to precisely identify ABO blood subgroup. Individuals with the same genotype would show different serological characteristics.

Objective Although the epidemic of coronavirus disease 2019 (COVID-19) has been controlled in China, large-scale testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still needed. Therefore, it is necessary to establish a method that can detect the new coronavirus on a large scale and improve the sensitivity and accuracy of the new coronavirus detection. Methods The dilution of mixed multiple samples could cause the low copy number of the virus and result in the inaccurate detection. The nested PCR coupled with qRT-PCR was designed to improve the sensitivity and accuracy of detection. The copy number of the tested sample was greatly increased by external primer PCR. The N gene sequence recommended by the National Center for Disease Control and Prevention (CDC) was then detected by fluorescent quantitative PCR. Results The false-positive rate of detection was reduced, and the detection sensitivity was increased by 3 orders of magnitude on the original basis, and a single copy of the virus template could be stably detected. Conclusion This Nested-qRT-PCR can greatly improve the sensitivity of new coronavirus detection and provide a feasible method for large-scale detection of mixed multiple samples. 

Objective To analyze the expression of Keratin 17 (KRT17) in colorectal cancer (CRC) and its clinical prognostic value. Methods Bioinformatics methods were employed to analyze the expression of KRT17 in the CRC database and its prognostic value. Furthermore, the expression level of KRT17 protein in 95 pairs of CRC samples was evaluated by immunohistochemistry (IHC), and the relationships between the KRT17 expression and clinicopathological features and prognosis of patients with CRC were investigated. Results Bioinformatics analysis revealed that the expression level of KRT17 in CRC tissues was significantly higher than that in normal tissues. The expression of KRT17 was correlated with tumor pathological type, lymph node metastasis, and TNM stage, and its overexpression predicted poor overall survival (allP< 0. 05). IHC showed that the expression of KRT17 protein was significantly up-regulated in CRC tissues as compared with that in adjacent-tumor tissues (t = 45. 704, P< 0. 001), which was consistent with the results from bioinformatics analysis. High expression of KRT17 was significantly correlated with tumor grade, depth of tumor invasion, lymph node metastasis, TNM stage, lymphovascular invasion, and perineural invasion (all P< 0. 05). Moreover, the survival analysis demonstrated that the overall survival of CRC patients with high expression of KRT17 was significantly lower than that of CRC patients with low expression of KRT17 (χ 2 = 8. 510, P = 0. 004), and further univariate and multivariate analyses indicated that high expression of KRT17 was an independent risk factor for overall survival of CRC patients (HR: 2. 015, 95 % CI: 1. 219-4. 457, P = 0. 018). Conclusion KRT17 is significantly up-regulated in CRC tissues, and its high expression predicts tumor progression and poor prognosis. KRT17 is expected to become a promising biomarker for prognosis evaluation of CRC patients.

Objective To investigate the role and regulatory mechanismof kinesin family member 18B (KIF18B) in proliferation and migration of gastric cancer (GC) cells. Methods The expression of KIF18B in GC tissues and cells (MKN-28, AGS and HGC-27) was detected by using qRT-PCR and Western blotting. Cell proliferation was evaluated by CCK-8 and colony formation assays. Wound healing method was used to detect cell migration. The expression levels of E-cadherin, N-cadherin and MMP-3 were determined by Western blotting. Results The KIF18B expression was up-regulated in GC tissues and cells. Silencing KIF18B dramatically reduced the proliferation, colony formation capability and migration rate of AGS cells. The expression level of E-cadherin was increased, and the N-cadherin and MMP-3 expression levels were decreased. Moreover, the KIF18B knockdown significantly inhibited the CDCA8 expression but promoted the expression of p53 protein. Overexpression of CDCA8 could reverse the effects of KIF18B silencing on cell proliferation and migration. Conclusion KIF18B regulated the proliferation and migration of GC cells via the CDCA8 / p53 pathway.

Objective To study the effect of total flavonoids (TF) of Melastoma dodecandrum on insulin resistance and renal damage in rats with diabetic nephropathy (DN). Methods Ten of 60 rats were randomly selected as normal group, and the remaining 50 rats were intraperitoneally injected with streptozotocin to establish the DN model and finally 43 DN models were successfully established. The 40 rats with DN were selected and randomly divided into four groups (n = 10 each), including DN model group, low-dose TF [150 mg / (kg·d)] group, middle-dose TF [300 mg / (kg·d)] group and high-dose TF [600 mg / ( kg·d)] group. On the 2nd day after the establishment of the rat DN models, the animals in TF-treated groups were given TF at different doses by gavage, and those in control group and model group were given the same volume of normal saline. The treatments lasted 30 d, and then renal tissues were removed for pathological analysis. The laboratory indicators including fasting plasma glucose (FPG), fasting insulin (FINS), serum creatinine (Scr), blood urea nitrogen (BUN), uric acid (UA) and microalbumin ( mAlb) were determined. The insulin resistance index (HOMA-IR) and the sensitivity index (ISI) were calculated. Results The levels of FPG, FINS, Scr, BUN, UA and mAlb in DN model group were significantly higher than those in control group (P< 0. 05). The FPG, FINS and HOMA-IR were significantly reduced (P < 0. 05) while ISI was significantly increased in TF-treated groups as compared with those in DN model group (P < 0. 05), and the changes of HOMA-IR and ISI were increased significantly with the increase of TF dose (P < 0. 05). The levels of Scr, BUN, UA and mAlb were significantly lowered in a dose dependent manner in TF-treated groups compared to those in DN model group (P < 0. 05). Conclusion TF of Melastoma dodecandrum can effectively improve the IR, decrease the blood glucose, alleviate the pathological damage of renal tissues and promote the recovery of renal function of DN rats.

Objective To investigate the effects of Reduning injection on lung function injury and immune disorder induced by Klebsiella pneumoniaein young mice. Methods The mice were randomly divided into 5 groups: control group, model group and low-, medium-and high-dose Reduning injection groups. HE staining was used to detect the pathology of lung tissues. Lung histopathological score and W/ D ratio were used to assess lung injury. The expression of Ki67 and Survivin mRNA was detected by RT-PCR. The expression of Caspase-3, Caspase-9, TGF-β and α-SMA proteins were detected by Western blot. Masson staining was used to determine pulmonary fibrosis. The contents of iNOS and IL-10 in lung tissue and peripheral blood were determined by ELISA. Results In Reduning injection groups relative to the KPN group, the lung injury and pulmonary fibrosis were significantly improved; the lung tissue score and W/ D ratio were markedly decreased (P < 0. 05) . The expression levels of Ki67 and Survivin mRNA were significantly increased (P< 0. 05). The expression levels of Caspase-3, Caspase-9, TGF-β and α-SMA protein were profoundly decreased (P< 0. 05). The iNOS content in tissues and peripheral blood was significantly decreased ( P < 0. 05). The IL-10 content was obviously increased (P < 0. 05). Conclusion Reduning injection can alleviate the lung function injury and immune disorder caused byKlebsiella pneumoniaein young mice.

Sepsis is a common cause of death in intensive care units worldwide. Due to the complexity of this immune syndrome, there is an urgent need to develop new therapeutic strategies. Aquaporins (AQPs) are involved in multiple physiological functions in sepsis, and their expression is regulated to varying degrees. AQPs could be used as drug targets or biomarkers for sepsis treatment, as they are key regulators in sepsis. Further studies on the regulation mechanisms of AQPs may help identify potential therapeutic targets for sepsis.

Ischemia-reperfusion injury often occurs after skin flap transplantation, and may lead to partial or complete necrosis of the skin flap. However, the pathogenesis of flap ischemia reperfusion injury remains unclear. Previous studies imply that its pathogenesis may be related to cell apoptosis, inflammatory response, tissue microcirculation disorder, and cell oxidative damage. The treatments for reducing the flap ischemia-reperfusion injury in previous studies include the use of ischemic preconditioning, drug intervention, reduction of inflammatory response, transplantation of stem cells or stem cell-based gene therapy, extracorporeal shock wave, antioxidant response, and inhibition of cell apoptosis. This article reviews the recent experimental researches on skin flap ischemia-reperfusion injury treatment and their clinical applications, which aims to provide theoretical basis and new ideas for the treatment of skin flap ischemia-reperfusion injury.

Treacher Collins syndrome (TCS, OMIM number 154500) is an autosomal dominant genetic disease that results in congenital craniofacial malformation. The pathogenesis of TCS is complex. Treacle ribosome biogenesis factor 1 (TCOF1) is an original pathogenic gene of TCS. In this review, we summarized the structure and function of TCOF1 and its protein Treacle, as well as the mutations of TCOF1 in TCS. Moreover, we listed the molecular role of TCOF1 in several signal pathways which are implicated in the development of TCS. The pathways involve telomere replication, DNA damage response (DDR) and p53 pathway. Finally, the prospects of targeting TCOF1 for TCS treatment were discussed.