OBJECTIVETo construct mouse enhanced green fluorecence protein (EGFP) -peroxisome proliferator-activated receptor (PPAR)gamma2, and to detect EGFP-PPARgamma2 expression in infected mouse bone marrow mesenchymal stem cells (BMSC).

METHODSCut the fragment of PPARgamma2 from the expression plasmid pcDNA flag PPARgamma2, then cloned the gene fragment into pEGFP-C1 and pEGFP-N1 vector. Subsequently, subclone the fragment EGFP-PPARgamma2 from pEGFP-C1-PPARgamma2 into the shuttle plasmid DC315. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid DC315-EGFP-PPARgamma2 and large adenovirus helper plasmid pBHGlox deltaE1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus Ad-EGFP-PPARgamma2 was confirmed. Then it was propagated in HEK293 cells. After the BMSC were transfected for 72 h, adipogenic differentiation was demonstrated.

RESULTSHEK293 cells were transfected with the pEGFP-C1-PPARgamma2 or pEGFP-N1-PPARgamma2 in mediation of liposome. The former green fluorescence protein was better than the latter by fluorescence microscope. The recombinant plasmids were digested and identified. Western blot analysis showed the expression of EGFP-PPARgamma2 in vitro. EGFP-PPARgamma2 protein was detectable in the nucleus of BMSC.

CONCLUSIONThe recombinant adenovirus encoding EGFP-PPARgamma2 fusion protein was successfully constructed, which provided a basis for application of EGFP-PPARgamma2 gene to adenovirus-mediated gene therapy.

Adenoviridae ; Animals ; Bone Marrow Cells ; metabolism ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Mice ; PPAR gamma ; metabolism ; Recombinant Proteins ; metabolism ; Transfection

Objective:To investigate the role of metformin (MET) in inflammatory response in abdominal aortic aneurysm(AAA) formation induced by angiotensin Ⅱ.Methods:AAA models were established by Ang Ⅱ infusion in ApoE-/-mice.Thirty male ApoE-/-mice (8-10-week-old) were randomly and equally divided into four groups:Control group,AAA group and AAA+MET group.HE staining and immunohistochemical staining were used to analyze the inflammatory response.Real-time quantitative polymerase chain reaction (qPCR) was used to analyze the mRNA expression of intercellular cell adhesion molecule-1 (ICAM-1).Western blotting was used to analyze the activation of nuclear factor κB signaling (NF-κB).Results:The AAA incidence and mean maximum suprarenal aortic diameter of AAA group is larger than the AAA+MET group.Compared with the AAA grouP,the expression of ICAM-1 and activity of NF-κB is lower in AAA+MET group.Conclusion:Metformin can attenuate AAA formation partially by regulating macrophage infiltration through regulating the mRNA expression of ICAM-1 regulated by the activation of NF-κB signaling pathway.

BACKGROUND: Activation of CD40 pathway negatively regulates the therapeutic effect of endothelial progenitor cells (EPCs), while inhibition of this pathway can enhance the biological function of the cells. OBJECTIVE: To compare the therapeutic effects of CD40-silenced EPCs (EPCs shRNA-CD40) and conventional EPCs transplantation in an animal model of pulmonary hypertension, and to explore the interventional effect of astragalosides on EPCs transplantation in the treatment of pulmonary hypertension. METHODS: Ninety SD rats were randomly divided into five groups: normal control group (n=24), model group (n=24), lentivirus transfection group (n=18), conventional transplantation group (n=18) and astragaloside group (n=6). Except the normal control group, the remaining four groups were given monocrotaline to induce pulmonary hypertension. Rats in the lentivirus transfection and conventional transplantation groups were given intravenous injection of Lv-shRNA-CD40-transfected EPCs and conventional EPCs respectively at 7, 14, 21 days after modeling (n=6 at each time point). Rats in the astragaloside group were given daily intraperitoneal injection of 80 mg/(kg?d) astragaloside within 1-21 days after modeling, and then Lv-shRNA-CD40-transfected EPCs were intravenously injected at 21 days after modeling. Hemodynamics, plasma endothelin-1 level and right ventricular hypertrophy index were detected at 28 days after modeling. RESULTS AND CONCLUSION: (1) After modeling, right ventricular pressure, mean pulmonary arterial pressure and right ventricular hypertrophy index were all increased compared with the normal control group (P < 0.05), while these indices were then decreased significantly after EPCs transplantation (P < 0.05). With the increasing of transplantation time, there was an increasing trend in the right ventricular pressure, mean pulmonary arterial pressure and right ventricular hypertrophy index in the two EPCs transplantation groups, but this trend was not remarkable in the lentivirus transfection group. (2) After modeling, the level of endothelin-1 was increased significantly compared with the normal control group (P < 0.05), and then decreased after EPCs transplantation (P < 0.05). The level of endothelin-1 in the lentivirus transfection group was significantly lower than that in the conventional transplantation group at the same time point (P < 0.05). (3) A significant improvement in hemodynamics, plasma endothelin-1 level and right ventricular hypertrophy index was observed in the astragaloside group as compared with the lentivirus transfection group (P < 0.05). Given the above findings, CD40-silenced EPCs transplantation is more effective and durable than the conventional transplantation in the treatment of pulmonary hypertension, and moreover, astragaloside can enhance the therapeutic effect.

Objective:To evaluate our novel path based on anatomical division of the anterior lateral wall of calcaneus and the sustentaculum tali for precise sustentacular screw placement in the surgical treatment of calcaneal fractures of Sanders types Ⅱ and Ⅲ.Methods:The anterior lateral wall of the calcaneus was divided into the anterior-superior zone S 1, the anterior-inferior zone S 2, the posterior-superior zone S 3 and the posterior-inferior zone S 4 for demarcation of the screw insertion points by our method of Four Zones, and into the front, middle and rear sections by our method of Three Sections for demarcation of the screw target points. The specimens were scanned by CT and modeled by Mimics. On the 3D virtual model of the calcaneus, one screw was placed from each zone of the anterior lateral wall of the calcaneus to the sustentaculum tali body. The screw placement target for S 1 and S 2 was the medial intersection point P 1 of the front and middle sections of the sustentaculum tali, and that for S 3 and S 4 was the medial intersection point P 2 of the middle and rear sections of the sustentaculum tali. It was observed whether the screws were placed in the bone channel. A total of 72 patients were included who had been admitted to Department of Orthopaedics, Suqian Hospital Affiliated to Xuzhou Medical University for calcaneal fractures of Sanders types Ⅱ and Ⅲ from January 2017 to January 2021. They were divided into an anatomical division group and a 3D printing group according to their screw placement method for the sustentaculum tali. In the anatomical division group of 32 patients subjected to screw placement based on our anatomical division, there were 25 males and 7 females, aged from 24 to 60 years; in the 3D printing group of 40 patients subjected to screw placement assisted by 3D printing, there were 31 males and 9 females, aged from 25 to 58 years. The disparities between the parameters of sustentacular screw placement and the actual values were compared in the anatomical division group, and the total number of screws, screws on average, distribution of screws, and accuracy of screw placement were compared between the 2 groups. Results:All the screws which were virtually placed in the specimens of the calcaneus from S 1 and S 2 to P 1 and from S 3 and S 4 to P 2 passed through the bony channel, with no perforation into the tarsal sinus. There was no significant difference in the general date between the anatomical division group and the 3D printing group, showing they were comparable ( P > 0.05). In the anatomical division group, a total of 52 screws were placed to the sustentaculum tali with an average of (1.63 ± 0.48) screws per patient, and 2 screws were placed in 20 patients, yielding an accuracy rate of screw placement of 92.3% (48/52). There were no statistically significant differences between the parameters and the actual values of screw placement in the anatomical division group ( P > 0.05). In the 3D printing group, a total of 63 screws were placed to the sustentaculum tali with an average of (1.58 ± 0.49) screws per patient, and 2 screws were placed in 23 patients, yielding an accuracy rate of screw placement of 93.7% (59/63). There were no significant differences in the above comparisons between the anatomical division group and the 3D printing group ( P > 0.05). Conclusion:In the surgical treatment of calcaneal fractures of Sanders types Ⅱ and Ⅲ, the sustentacular screw placement based on our anatomical division of the anterior lateral wall of the calcaneus and the sustentaculum tali can lead to similar clinical accuracy as 3D printing-assisted screw placement does.

The development of new biomarkers or therapeutic targets for cancer immunotherapies requires deep under-standing of T cells. To date, the complete landscape and systematic characterization of long noncoding RNAs (lncRNAs) in T cells in cancer immunity are lacking. Here, by systematically analyzing full-length single-cell RNA sequencing (scRNA-seq) data of more than 20,000 libraries of T cells across three cancer types, we provided the first comprehensive catalog and the functional repertoires of lncRNAs in human T cells. Specifically, we developed a custom pipeline for de novo transcriptome assembly and obtained a novel lncRNA catalog containing 9433 genes. This increased the number of current human lncRNA catalog by 16％and nearly doubled the number of lncRNAs expressed in T cells. We found that a portion of expressed genes in single T cells were lncRNAs which had been overlooked by the majority of previous studies. Based on metacell maps constructed by the MetaCell algorithm that partitions scRNA-seq datasets into disjointed and homogenous groups of cells (metacells), 154 signature lncRNA genes were identified. They were associated with effector, exhausted, and regulatory T cell states. Moreover, 84 of them were functionally annotated based on the co-expression networks, indicating that lncRNAs might broadly participate in the regulation of T cell functions. Our findings provide a new point of view and resource for investigating the mechanisms of T cell regulation in cancer immunity as well as for novel cancer-immune biomarker development and cancer immunotherapies.

Coronavirus disease 2019(COVID-19),which is caused by SARS-CoV-2,varies with regard to symptoms and mortality rates among populations.Humoral immunity plays critical roles in SARS-CoV-2 infection and recovery from COVID-19.However,differences in immune responses and clinical features among COVID-19 patients remain largely unknown.Here,we report a database for COVID-19-specific IgG/IgM immune responses and clinical parameters(named COVID-ONE-hi).COVID-ONE-hi is based on the data that contain the IgG/IgM responses to 24 full-length/truncated proteins corresponding to 20 of 28 known SARS-CoV-2 proteins and 199 spike protein peptides against 2360 serum samples collected from 783 COVID-19 patients.In addition,96 clinical parameters for the 2360 serum samples and basic information for the 783 patients are integrated into the database.Furthermore,COVID-ONE-hi provides a dashboard for defining samples and a one-click analysis pipeline for a single group or paired groups.A set of samples of interest is easily defined by adjusting the scale bars of a variety of parameters.After the"START"button is clicked,one can readily obtain a comprehensive analysis report for further interpretation.COVID-ONE-hi is freely available at www.COVID-ONE.cn.

The bone morphogenetic protein (BMP) family is an important factor in the regulation of cell ular life activities and in the development of almost all tissues. BMP-mediated signaling plays an important role in tooth root development, which is a part of tooth development. Epithelial and mesenchymal interactions are involved in tooth root development, but the BMP signaling pathway has a different effect on tooth root development in epithelial and mesenchymal. This review summarizes the advances of BMP signaling in tooth root development.
Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein 7 ; Bone Morphogenetic Proteins ; physiology ; Odontogenesis ; Signal Transduction ; Tooth ; Tooth Root ; growth & development

The polarity of ameloblasts and odontoblasts is crucial for their differentiation and function. Polarity-related molecules play an important role in this process. This review summarizes the process of polarity formation of ameloblasts and odontoblasts and their related regulators.
Ameloblasts ; Cell Differentiation ; Odontoblasts

OBJECTIVE: Identification of new risk factors is needed to improve prediction of adverse outcomes in patients with three-vessel disease (TVD). The present study aimed to evaluate the prognostic values of serum chloride and sodium levels in patients with TVD. METHODS: We used data from a prospective cohort of consecutive patients with angiographically confirmed TVD. The primary endpoint was all-cause death. Cox proportional hazard regression was used to analyze the relationship of serum chloride and sodium levels with long-term outcomes of TVD patients. RESULTS: A total of 8,318 participants with available serum chloride and sodium data were included in this analysis. At baseline, patients in the low tertiles group of serum chloride level (⪕ 102.0 mmol/L) or serum sodium level (⪕ 139.0 mmol/L) had more severe disease conditions. During a median follow-up of 7.5-year, both low serum chloride level and low serum sodium level were found to be associated with an increased risk for mortality in univariate analysis. However, when both parameters were incorporated into a multivariate model, only low serum sodium level remained to be an independent predictor of all-cause death (hazard ratio: 1.16, 95% confidence interval: 1.01-1.34, P = 0.041). Modest but significant improvement of discrimination was observed after incorporating serum sodium level into the Synergy between percutaneous coronary intervention (PCI) with Taxus and Cardiac Surgery score. CONCLUSION Serum sodium level is more strongly associated with long-term outcomes of TVD patients compared with serum chloride level. Low serum sodium level is an independent risk factor for mortality, but only provides modest prognostic information beyond an established risk model.
Aged ; China ; epidemiology ; Chlorides ; blood ; Coronary Artery Disease ; blood ; diagnosis ; mortality ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Prospective Studies ; Sodium ; blood