Animals ; Antineoplastic Agents ; therapeutic use ; Gene Silencing ; Genetic Therapy ; methods ; Humans ; MicroRNAs ; antagonists & inhibitors ; therapeutic use ; Neoplasms ; genetics ; therapy ; RNA Interference ; physiology ; RNA, Small Interfering ; antagonists & inhibitors ; therapeutic use ; RNA-Induced Silencing Complex ; metabolism

Congenital Hyperinsulinism ; diagnosis ; Humans ; Infant, Newborn ; Male

Purpose:To investigate the relationship between cell apoptosis and dephosphorylated RB protein in human breast cancer. Methods:In our work,human breast cell lines (MCF-7/S,the chemosensitive cell line and MCF-7/ADR,the chemoresistent cell line)were evaluated. Chemosensitivity of two cell lines was evaluated by the MTT colorimetric assay;the expressive levels of dephosphorylated RB protein were detected with immunocytochemistry. Apoptosis rates were determined by flow cytometry(FCM). Results:ADR inhibited proliferation of chemosensitive cell line MCF-7/S ,the 50% inhibition concentration (IC 50 ) was 0.128 ?g/ml;And IC 50 of MCF-7/ADR was 10.89 ?g/ml. The chemotherapeutic sensitivity of MCF-7/S was more than that of MCF-7/ADR by 86 times . Before treatment with ADR,phosphorylated RB protein was positive in two cell lines,but dephosphorylated RB protein was negative;After treatment of different concentration ADR,when the concentration of ADR was increased,expression of dephosphorylated RB protein elevated accordingly in MCF-7/S,but no significant change in MCF-7/ADR. Apoptosis and cell cycle was detected by FCM assays shows ADR induced apoptosis of MCF-7/S more than MCF-7/ADR(P0.05).

Objective To evaluate the security and biocompatibility of a new kind of absorbable high-strength PDLLA-CPP composite, which is used for internal fracture fixation. Methods According to the national standard of the procedure of appraise in food security and toxicology, micronucleus test of bone marrow cell, spermatic aberration test, chromosomal aberration test, and teratogenic test were measured with IP maximal tolerance dose of maceration extract from PDLLA-CPP composites. Meanwhile, chondrocyte culture with PDLLA-CPP composite was used to evaluate the effect of PDLLA-CPP composite on cartilage cell proliferation. Results Micronucleus test of bone marrow cell, spermatic aberration test, chromosomal aberration test, and teratogenic test were not affected by IP maximal tolerance dose of PDLLA-CPP maceration extract. PDLLA-CPP composite also had no cytotoxicity on cartilage cell proliferation. Conclusion PDLLA-CPP is a new and safe medical material with good biocompatibility and absorbability.

Objective To study the feasibility and effect of implantation of the biliary stent for treatment of biliary obstruction.Methods A retrospective analysis of the diagnostic and therapeutic procedure was done in 131 cases of the inpatients from Apr.2006 to Feb.2007.The 131 patients with biliary obstruction underwent successfully 138 cases/times implantation of biliary stent.The results of recession of jaudince and the rate of complication were evaluated.Results All patients were implanted biliary stent successfully.The serum total bilirubin,direct bilirubin,alkaline phosphatase,?-glutamyl transpeptidase decreased obviously in three days,and the difference was remarkable.The main complication was infection of biliary tract and pancreatitis.Conclusion Implantation of biliary stent is an effective management for biliary obstruction,especially for patients who have lost the chance of operation.

Isoliquiritigenin (ISL) is a flavonoid compound isolated from licorice. It possesses excellent antioxidant and anti-diabetic activities. This study aims to investigate the molecular mechanism underlying the alleviatory effect of ISL on energy metabolism imbalance caused by type 2 diabetes mellitus (T2DM). 8-week-old male C57BL/6J mice were used in in vivo experiments. The high-fat-high-glucose diet combined with intraperitoneal injection of streptozotocin was applied to establish T2DM animal model. All animal experiments were performed in accordance with the Institutional Guidelines of Laboratory Animal Administration issued by the Committee of Ethics at Beijing University of Chinese Medicine. HepG2 cells were used in in vitro experiments. Enzyme-linked immunosorbent assay (ELISA) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to examine the protein and mRNA levels of mitochondrial function-related targets. The levels of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) in HepG2 cells were measured by the flow cytometry. Additionally, the molecular docking of ISL and key target proteins was analyzed. It was found that ISL significantly inhibited the activity of mitochondrial respiratory chain complex I and increased the protein levels of uncoupling protein 2 (UCP2) in the livers of mice and HepG2 cells. It also obviously decreased the ROS levels and increased the MMP levels in cultured HepG2 cells. In addition, ISL promoted mitochondrial biogenesis by activating proliferator-activated receptor gamma co-activator 1α (PGC-1α) and enhanced mitophagy by upregulating Parkin. It also improved mitochondrial fusion by increasing the mRNA and protein levels of mitofusin 2 (MFN2). In conclusion, ISL alleviates energy metabolism imbalance caused by T2DM through suppression of excessive mitochondrial oxidative phosphorylation and promotion of mitochondrial biogenesis, mitophagy, and fusion.

Isoliquiritigenin (ISL) is an active chalcone compound isolated from licorice. It possesses anti-inflammatory and anti-oxidative activities. In our previous study, we uncovered a great potential of ISL in treatment of type 2 diabetes mellitus (T2DM). Therefore, this study aims to reveal the mechanism underlying the alleviatory effects of ISL on T2DM-induced glycolipid metabolism disorder. High-fat-high-sugar diet (HFD) combined with intraperitoneal injection of streptozotocin (STZ) were used to establish T2DM mice model. All animal experiments were carried out with approval of the Committee of Ethics at Beijing University of Chinese Medicine. HepG2 cells were used in in vitro experiments, and sodium palmitate (SP) was applied to establish insulin resistance (IR) model cells. The effects of ISL on body weight, fasting blood glucose levels, and pathological changes in the livers of mice were examined. Enzyme-linked immune sorbent assay (ELISA) and real-time quantitative PCR (RT-qPCR) were applied to detect the regulatory effects of ISL on key targets involved in glucolipid metabolism. Additionally, molecular docking and analytical dynamics simulation methods were used to analyze the interaction between ISL and key target protein. The results indicate that ISL significantly downregulates the transcriptional levels and inhibits the activities of key enzymes involved in gluconeogenesis, including pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK), and fructose-1, 6-bisphosphatase (FBP). It also downregulates the transcriptional and protein levels of hepatocyte nuclear factor 4α (HNF4α) and cAMP response element binding protein (CREB), the two transcriptional factors involved in gluconeogenesis. Thus, ISL inhibits hepatic gluconeogenesis in T2DM mice. In addition, ISL reduces total cholesterol (TC) and triglyceride (TG) levels in the livers of T2DM mice. Moreover, ISL downregulates the mRNA levels of lipogenesis genes and upregulates those of genes involved in fatty acid oxidation, lipid uptake, and lipid export. In conclusion, ISL suppresses hepatic gluconeogenesis, promotes lipolysis, and restrains lipogenesis in T2DM mice, thereby improving the abnormal glycolipid metabolism caused by T2DM.

The aim was to construct and identify the mammalian expression vector of pCAG-IRES-SHIP-GFP and to detect whether it could express in human acute leukemia cell line K562.The cDNA fragment of SHIP obtained by RT-PCR was inserted into pCAG-IRES-GFP.The recombinant plasmid was confirmed by restriction enzyme digesiton,PCR and DNA sequecing.pCAG-IRES-SHIP-GFP was transfected into K562 cells with lipofectamine 2000.The expression of SHIP was determined by GFP fluorescence and Western blot analysis.FQ-PCR was used to quantitate SHIP mRNA.The expression of p-Akt,Akt were determined by Western blot.PI were tested by flow cytometry and MTT to verify whether exogenous SHIP could inhibit proliferation of K562 cells.The results showed that the correct constrution of the recombinant plasmid pCAG-IRES-SHIP-GFP has been shown by restriction enzyme digestion,PCR and DNA sequencing.pCAG-IRES-SHIP-GFP could express SHIP protein in K562 cells.The K562 cells viability after transfected with SHIP gene droped down.Western blot analysis showed that phospha-Akt308 and Akt473 were reduced to 38.7% and 68% respectively.It was concluded that the vector of pCAG-IRES-SHIP-GFP has been successfully constructed and it can be expressed in K562 cells.The expression of exogenous SHIP gene can lead to apoptosis of K562 cells by down-regulating the p-Akt expression.What found here might be one of the mechanisms involved in the pathogenesis of leukemia.

A BBB co-culture cell model consisting of rat brain microvascular endothelial cells (BMEC) and astrocytes (AS) was established to study the effect of Angelica dahurica coumarins on the transport behavior of puerarin across blood-brain barrier (BBB) in vitro and in vivo. The barrier function of this model was evaluated by measuring the transendothelial resistance, phenol red permeability and BBB related protein expression. The permeability assay and western blot methods were performed to study the effects of Angelica dahurica coumarins on the BBB permeability and the expression of BBB related protein. The animal experiment protocols in this study were approved by the Animal Ethics Committee of Xi'an Jiaotong University (Animal Ethics No.: 2021-1329). The results showed that the established BMEC/AS co-culture model could be used to evaluate drug transport across BBB in vitro. After combined with Angelica dahurica coumarins, the transport capacity of puerarin was significantly increased in vitro and in vivo. Additionally, Angelica dahurica coumarins enhanced BBB permeability and inhibited the protein expression of P-glycoprotein (P-gp), zonula occludens-1 (ZO-1) and occludin. Angelica dahurica coumarins might increase BBB permeability by inhibiting the expression of P-gp and tight junction protein, thereby increasing the content of puerarin in brain tissue.

Objective To study the clinical outcomes of robotic extended thymectomy and thoracoscopic extended thymectomy for thymoma patients with myasthenia gravis compared with conventional median sternotomy extended thymectomy.Methods The clinical data of thymoma patients with myasthenia gravis treated by extended thymectomy between June 2013 and June 2016 were retrospectively reviewed.The clinical outcome parameters were compared according to surgical approach.Results 41 thymoma patients with myasthenia gravis,8 cases underwent robotic extended thymecotmy,11 cases underwent thoracoscopic extended thymectomy and 20 underwent median sternotomy extended thymectomy.The resected extension included tumor,thymus tissue and adipose tissue in anterior mediastinum.There were no significant differences between robotic group and thoracoscopic group regarding operative time,blood loss,chest tube duration,hospital stay,postoperative complications and postoperative myasthenic crisis (P ＞ 0.05).The blood loss of robotic group and thoracoscopic group was significantly lower than that in median sternotomy group(P ＜ 0.05).The chest tube duration of thoracoscopic group was significantly shorter than that in median sternotomy group(P ＜0.05).The effective rates of MG after extended thymectomy in robotic group,thoracoscopic group and sternotomy group was 65.0％ 、69.2％ 、62.5％ respectively and there was no significant difference (P ＜ 0.05).Conclusion Robotic thymectomy and thoracoscopic thymecotomy are both minimal invasive surgery approach with less bleeding for thymoma patients with myasthenia gravis.The clinical outcomes of robotic thymectomy and thoracoscopic thymecotomy are similar.