Objective To construct and authenticate the lentiviral-mediated overexpression of mouse mitochondrial-targeted-8-oxoguanine DNA-glycosylase 1 (mito-OGG1) gene and the lentiviral-mediated short hairpin RNA (shRNA) down-regulation of OGG1 gene expression model in 661W cells.Methods Constructed the target plasmids,including pLenti-EF1a-EGFP-P2A-Puro-CMV-Mito-OGG1-3Flag (pLenti-OGG1-GFP) and pLKD-CMV-G&PR-U6-shRNA (pLKD-shRNA).293T cells were used to obtain green fluorescent protein (GFP)-tagged lentiviral vector of interest by using a second generation lentivirus packaging system.293T cells were also used for the virus titer estimation.The multiplicity of infection (MOI) of 661W cells was detected by fluorescence microscopy.A stable transfected cell line was screened by puromycin.Immunofluorescence was used to detect transfection efficiency and cytochrome C oxidase Ⅳ (COXⅣ)-OGG1 co-localization.OGG1 mRNA and protein expression levels were detected by real-time qantitative PCR (QPCR) and Western blot.Results Sequencing results showed that the inserted sequence in the over-expression plasmid was consistent with the mouse OGG1 (NM_010957.4) gene sequence in the gene library.The original lentiviral titer after packaging and purification was between 2.0× 107to 6.0× 107 TU/ml.The optimal MOI of 661W cells was 40,and puromycin with a concentration of 4.0 μg/ml successfully screened stable transformation.The transfection efficiency was up to 100％ after screening.Immunofluorescence demonstrated successful co-localization of OGG1 and COXⅣ.The relative expression levels of OGG1 mRNA in the blank control group,OGG1 group,overexpression control group,shRNA group and low expression control group were 1.000±0.000,41.581±12.206,0.888±0.056,0.239±0.121 and 1.081±0.083,and the relative expression levels of OGG1 protein were 1.029±0.153,1.657 ± 0.237,0.752 ± 0.143,0.471 ± 0.149 and 1.036 ± 0.185,respectively,with significant differences between them (F=44.654,30.948;both at P＜0.05),the relative expression levels of OGG1 mRNA and protein in the OGG1 group were significantly higher than those in the overexpression control group,the relative expression levels of OGG1 mRNA and protein in the shRNA group were significantly lower than those in the lower expression control group,with significant differences between them (all at P＜0.05).Conclusions The mitoOGG1 overexpression and OGG1 knockdown models of 661W cells are successfully constructed,which provides the preliminary experimental basis for follow-up study.

Objective: To investigate the correlation between the expression of E2F1 and growth arrest and DNA damage inducible protein 45g (GADD45g) in patients with acute myeloid leukemia (AML), and to explore whether GADD45g exerts its induction of DNA damage, cell apoptosis, senescence, cell cycle arrest and drug sensitivity in AML through inhibition of E2F1. Methods: A total of 32 cases of bone marrow specimens from patients initially diagnosed asAML in Hospital of Blood DiseasesAffiliated to ChineseAcademy of Medical Sciences from January 2013 to December 2016, were selected for this study; In addition, AML cell lines (U937, HL60, THP-1, Molm-13) were also collected for this study. The mRNAexpression of GADD45g and E2F1 in above mentioned specimens and cell lines by qPCR,andtheircorrelationwasalsoanalyzed.Thelentiviral vector over-expressing E2F1 (pLV-E2F1-RFP) was constructed to prepare recombinant lentivirus, which was then transfected Molm-13 and THP-1 cells that over-expressing GADD45g. Whether GADD45g exerts tumor inhibition effect on AML cells through inhibition of E2F1 was determined by comet assay, Annexin V/7AAD flow cytometry, β-galactosidase staining and PI staining flow cytometry etc. Results: The mRNA expression of GADD45g was negatively correlated with E2F1 in bone marrow of AML patients and AML cell lines (r=–0.663, P<0.01). Over-expression of GADD45g significantly inhibited the expression of E2F1 in AML cell lines (all P<0.01). Molm-13 and THP-1 cells that simultaneously over-expressing GADD45g and E2F1 were successfully constructed. Compared with the control group, the protein expressions of GADD45g and E2F1 in over-expression groups were significantly increased (all P<0.01). Compared with cells over-expressing GADD45g alone, simultaneous over-expression of both GADD45g and E2F1 significantly reduced the apoptosis, senescence and DNA damage (all P< 0.01), and rescued cell cycle arrest in Molm-13 and THP-1 cells (all P<0.01), thus further reduced the chemo-sensitivity of AML cells caused by GADD45g over-expression (all P<0.01). Conclusion: GADD45g exerts anti-tumor effect inAMLvia inhibition of E2F1.

Objective: To study the clinicopathological features, immunohistochemical phenotype, molecular changes, differential diagnosis and prognosis of eosinophilic solid and cystic renal cell carcinoma (ESC RCC). Methods: A total of 15 cases were selected from 2005 to 2019 at Nanjing Jinling Hospital,Nanjing University School of Medicine for clinicopathological and immunohistochemical analysis, 10 of which were subject to cancer-associated mutation analysis using targeted next-generation sequencing (NGS) panel. A literature review was also performed. Results: The patients′ ages ranged from 15 to 68 years (mean, 33 years). The male-to-female ratio was 1.1∶1.0. During a mean follow-up of 22 months, none of the patients developed tumor recurrence, progression or metastasis. Histologically, the tumors typically demonstrated solid and cystic architectures and the neoplastic cells contained voluminous eosinophilic cytoplasm with prominent granular cytoplasmic stippling. Immunohistochemically, tumor cells in all cases were immunoreactive for CK20. Signal pathway related protein mTOR and S6 were positive in 14/15 and 6/15 cases, respectively. Cathepsin K, Melan A and HMB45 were at least focally positive in 12/15, 6/15 and 2/15 cases, respectively. CK7 and CD10 showed focal immunostain positivity in some cases, while TFE3, TFEB, CA9 and CD117 were negative in all cases. NGS demonstrated TSC1/TSC2 mutations in all tested cases (10/10). Conclusions ESC RCC is a rare tumor that tends to occur in young patients with an indolent behavior. Diagnosis can be established by its distinct clinical and histopathologic findings, immunohistochemical phenotype and molecular genetics. The tumor may be considered as a new subtype of RCC.

Objective To investigate the clinical efficacy and application value of submucosal tunneling endoscopic resection(STER)for upper gastrointestinal submucosal tumors(SMTs). Methods A retrospective analysis was performed on the endoscopic and clinical data of 44 cases with SMTs who received STER from January 2015 to June 2016 in Chinese PLA General Hospital. Results The rate of complete resection was 88.6%(39/44). The operating time was 60.1±30.6 min. The hospitalization time was 10.1± 3.3 days. The rate of complications was 6.8%(3/44). The diagnosis of SMTs by pathology and endoscopic ultrasonography(EUS),the size of SMTs measured by EUS and ruler after STER,and the growing direction judged by EUS and CT were consistent. Conclusion STER for SMTs has a higher complete resection rate, shorter operating time and hospitalization time, and fewer complications. EUS combined with CT is an effective method for preoperative evaluation.

Half a century ago,it was confirmed that leukemia could be induced by retroviruses in some animals.There are numerous en-dogenous retroviruses in the human genome.However,retrovirus-related human adult T-cell leukemia is only endemic in some local areas. Recently,an extensive study on apolipoprotein B mRNA editing enzyme catalytic polypeptide 3(APOBEC3)family members has revealed the resistance mechanism in human to retrovirus-induced leukemia.The Epstein-Barr virus is widely spread in the human population and exists as a latent infection in the human body for a long time.It is closely associated with the occurrence and development of some lymphomas and solid tumors,indicating its tumorigenicity.The relationship between other viral infections and leukemia/lymphoma remains unclear. The interaction may be mediated by complex,multistep,and indirect networks,which need to be further illuminated.

Soft tissue sarcomas (STSs) are malignant tumors originating from mesenchymal tissues. They have lower morbidity rates than other cancers, which is only about 1% among adult malignancies. STSs consist of several subtypes and many of them are highly malignant. The 5-year survival rate is approximately 50%-60%. Investigation of STS pathogenesis demonstrated that epigenetic chang-es were closely related to the occurrence and development of STSs. Different from genetics changes, epigenetic changes are heritable and occur mainly through DNA methylation, histone modification, non-coding RNA regulation, and chromatin remodeling, all of which occur without changing the DNA sequence. Elucidation of the epigenetic mechanisms involved in STSs can help develop new clinical di-agnosis and treatment methods, and this article aims to review the recent advances in epigenetic research on STS.

Protein nitration and nitrosylation are essential post-translational modifications (PTMs) involved in many fundamental cellular processes. Recent studies have revealed that excessive levels of nitration and nitrosylation in some critical proteins are linked to numerous chronic diseases. Therefore, the identification of substrates that undergo such modifications in a site-specific manner is an important research topic in the community and will provide candidates for targeted therapy. In this study, we aimed to develop a computational tool for predicting nitration and nitrosylation sites in proteins. We first constructed four types of encoding features, including positional amino acid distributions, sequence contextual dependencies, physicochemical properties, and position-specific scoring features, to represent the modified residues. Based on these encoding features, we established a predictor called DeepNitro using deep learning methods for predicting protein nitration and nitrosylation. Using n-fold cross-validation, our evaluation shows great AUC values for DeepNitro, 0.65 for tyrosine nitration, 0.80 for tryptophan nitration, and 0.70 for cysteine nitrosylation, respectively, demonstrating the robustness and reliability of our tool. Also, when tested in the independent dataset, DeepNitro is substantially superior to other similar tools with a 7%-42% improvement in the prediction performance. Taken together, the application of deep learning method and novel encoding schemes, especially the position-specific scoring feature, greatly improves the accuracy of nitration and nitrosylation site prediction and may facilitate the prediction of other PTM sites. DeepNitro is implemented in JAVA and PHP and is freely available for academic research at http://deepnitro.renlab.org.
Amino Acid Sequence ; Amino Acids ; metabolism ; Deep Learning ; Humans ; Internet ; Neural Networks (Computer) ; Nitrosation ; Proteins ; chemistry ; metabolism ; Reproducibility of Results ; Software

Objectives: To investigate the compensation of upper first molar in adult skeletal class Ⅲ patients. Methods: The sample consisted of 48 skeletal class Ⅲ patients with orthodontic-surgical treatment from February 2011 to January 2015 in Peking University School and Hospital of Stomatology. They were allocated into 2 groups based on extraction, 28 patients in extraction group and 22 patients in non-extraction group. Dental casts were taken before and after treatment, and all the casts were scanned with a computer-assisted three-dimensional (3D) scanning system. Acquired 3D data were analyzed by Rapidform 2006. The tip and torque of the upper first molar were measured and analyzed. Results: The tip and torque of the upper first molar in the skeletal class Ⅲ patients after treatment had no difference between extraction group and non-extraction group (P=0.150, P=0.144). The torque of upper first molar in extraction group [-3.32° (-5.10°, 1.16°)] before treatment was bigger than that in non-extraction group [-6.94° (-13.06°, -0.42°)] (P=0.005), and the inclination of upper first molar in extraction group was more obvious. Conclusions In skeletal class Ⅲ patients, the upper first molar inclined buccally to compensate the discrepancy and extraction was usually necessary for decompensation.

Human herpes virus (HHV) spreads widely in a latent-infected way. HHV is divided into 3 kinds and 8 types. Herpes simplex caused by HHV-1 and HHV-2, varicella-zoster caused by HHV-3 and Epstein-Barr virus (EBV) known as HHV-4 and recognized firstly as human tumor virus, are well known in the field of virology. With the application of hematopoietic stem cell transplantation , cytomegalovirus (HHV-5) and HHV-6 have gained an increasing attention. The biological characteristic of HHV is latent infection in a long time and HHV can be reactivated in some immunocompromised individuals. Clarification of latent infection mechanism will help provide targets for therapy and lay a foundation for clinical treatment. This paper summarizes the clinical significance of HHV infection. Taking the mechanism of EBV latent infection as an example, the pathway and significance as well as strategy of co-evolution of organism and virus will be discussed in order to provide clues for prevention and therapy.

Objective This study aims to analyze midterm survival and to assess the midterm mortality following acute kidney failure (AKF) of patients undergoing surgical treatment of acute type A aortic dissection.Methods Clinical data of the patients who underwent Sun's procedure from February 2009 to March 2015 were collected.Patients with preoperative dialysis were excluded.Data including preoperative cardiovascular diseases,serum creatinine leve1 and CPB time were gathered.The patients were divided into the group of AKF and the group of non-AKF accounting to require dialysis.The group of AKF was follow-up by phone.Midterm mortality was studied with Cox analysis and midterm survival was estimated by Kaplan-Meier analysis.Results 524 patients underwent aortic surgeries which AKF was occurred in 51 (51/524,9.7％) patients.Hospital mortality was 23.5％ (12/51) in AKF group which seven death were strokes,three death were lower-extremity ischemia and two death were low cardio output.There were 2 patients dead with multi organ failure and paraplegia in first year.One reoperation patient because of anastomotic fistula was dead in fifth year.Only diabetes was independently associated with midterm mortality.Midterm survival for patients with AKF was 56％ during a median follow-up of 30.5 months.Survival was equal between the subgroups of Cr ＞ 200 mol/L and Cr ＜ 200 mol/L(P =0.741).Conclusion AKF after acute aortic dissection was a severe complication and the incidence was 9.7％.In patients with AKF,diabetes was associated with increased mortality across follow-up.Five years survival was 56％ not effected by preoperative Cr.