OBJECTIVE To study the effect of Qing Yi Tang on acute pancreatitis(AP),especially AP complicated with endotoxemia and its possible mechanism.METHODS Fourty Wistar rats were divided into 5 groups including group A(the control group),group B(the AP group),group C(the AP group treated with Qing Yi Tang),group D(the AP group treated with LPS) and group E(the AP group treated with LPS + Qing Yi Tang).Pathological damage of pancreatic tissue was scored with HE staining.The mRNA expression of TNF-? was measured with semi-quantitative RT-PCR,and activation of NF-?B was detected with flow cytometry(FCM) assay.RESULTS It was shown in results that the expression of TNF-? mRNA,activation of NF-?B and pathological damage of the group B were all obviously higher than those of the group A.After treated with LPS which might promote the activation of NF-?B,there was seen the further rise of the activation of NF-?B,expression of TNF-? mRNA and pathological damage.When Qing Yi Tang intervention was applied,the activation of NF-?B and the expression of TNF-? mRNA could be remarkably relieved,so did the pathological damage of pancreas.CONCLUSIONS Qing Yi Tang may be applied to decrease activation of NF-?B and the expression of TNF-? so as to treat AP or AP with endotoxemia.

Objective To evaluate diagnostic value of pit pattern analysis on detection of early colorectal carcinoma. Methods 4176 patients were examined with colonoscopy and had the mucosal lesions stained with 0.4% indigo carmine, and part of them observed with magnifying endoscope and stereomicroscope, then compared the mucosal crypt patterns (the pit patterns Kudo classification) with pathologic diagnosis. Results There were 955 protruded and flat lesions on the large intestine mucosa in 752 patients, and among them there are 14 early cancers, 209 advanced cancers, 76Ⅱa、Ⅱb、Ⅱc、Ⅱa+Ⅱc lesions. We also found 43 laterally spreading tumors (LST) ranging from 16 to 110 mm in diameter, 2 for pit Ⅱ,18 for pit Ⅲ L, 19 for pit Ⅳ, 1 for pit Ⅴ A, 1 for Ⅴ N. The pit pattern of the most non neoplastic lesions was type Ⅰ or Ⅱ, which is about 85.4% (303/355), and the type of the adenomas was type Ⅲ or Ⅳ, about 86.0% (504/586). All the invasive carcinomas'pit patterns were type Ⅴ and there were 8 for type Ⅴ (2 Ⅴ A, 6Ⅴ N) among 14 early carcinomas. Conclusion Pit pattern analysis is a very important tool to determine the nature of lesions, which helps to decide the kinds of later therapeutic intervention.

OBJECTIVETo investigate the cytotoxic effect of multi-walled carbon nanotubes (MWCNTs) on human liver L02 cells and its relevant mechanism.

METHODSMWCNTs, carboxyl modification MWCNTs (MWCNTs-COOH), and hydroxyl modification MWCNTs (MWCNTs-OH) were characterized by transmission electron microscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy. The carbon nanotubes at concentrations of 12.5, 25, 50, 100, and 200 μg/ml were incubated with human liver L02 cells for 24, 48 and 72 hours, respectively. The cell viability was evaluated by water soluble tetrazolium salts assay and the intercellular reactive oxygen species induced by the carbon nanotubes were detected by 2', 7'-dichlorodihydrofluorescein diacetate method.

RESULTSTransmission electron microscope showed that the average outside diameters (10 to 20 nm) and the average length (10 to 30 μm) of the three MWCNTs were similar. Scanning electron microscope indicated that the three MWCNTs had a similar surface topography. X-ray photoelectron spectroscopy demonstrated that the MWCNTs-COOH and MWCNTs-OH had relatively high peak areas at 289 and 286ev, respectively,indicating that they have been modified by carboxyl and hydroxyl groups,respectively. Water soluble tetrazolium salts assay showed that the MWCNTs-COOH was less cytotoxic when compared to MWCNTs which demonstrated to be slightly more cytotoxic than MWCNTs-OH. The capability to induce increase in intracellular reactive oxygen species was in the following order: MWCNTs > MWCNTs-COOH > MWCNTs-OH.

CONCLUSIONSModification of MWCNTs with carboxyl group and hydroxyl group improves the biocompatibility of MWCNTs to some extents. MWCNTs-COOH has better compatibility than MWCNTs at the low concentration,and MWCNTs-OH showed better compatibility than MWCNTs after 48 hours. Different mechanisms may be involved in the interaction between cells and the MWCNTs with different chemical surfaces.

Cell Survival ; drug effects ; Cells, Cultured ; Hepatocytes ; drug effects ; metabolism ; Humans ; Nanotubes, Carbon ; chemistry ; toxicity ; Reactive Oxygen Species ; metabolism

OBJECTIVETo study the morphologic changes of collagen type III glomerulopathy and to investigate the possible cellular origin for collagen III production.

METHODSLight microscopy, immunofluorescent staining, immunohistochemistry (for collagen I, III and IV and alpha-SMA) and electron microscopy studies on 3 renal biopsy cases of collagen type III glomerulopathy were performed.

RESULTSTwo cases presented with nephrotic syndrome, one of which was associated with systemic hypertension. The third case showed renal impairment and renal hypertension. None had any known family history of renal diseases. Light microscopy showed diffuse thickened glomerular basement membrane and expanded mesangium with deposition of weakly PAS-positive homogeneous material not associated with mesangial cell proliferation. Electron microscopy revealed massive collagen fiber deposits in the subendothelial spaces and mesangium. The mesangial cells also contained bundles of microfilaments in the subplasmalemmal regions. Immunohistochemically, the diffuse positivity for type III collagen corresponded to the homogeneous material seen under light microscopy. The staining for type I and IV collagens was negative. Alpha-SMA was expressed in many mesangial cells.

CONCLUSIONSThe diagnosis of collagen type III glomerulopathy can be made on the basis of detailed morphologic examination and ancillary investigations. It is possible that activated mesangial cells may be the cellular origin of collagen III.

Actins ; metabolism ; Adult ; Collagen Type III ; metabolism ; Female ; Glomerular Basement Membrane ; pathology ; ultrastructure ; Glomerulonephritis ; metabolism ; pathology ; Humans ; Male ; Mesangial Cells ; metabolism ; pathology ; Middle Aged

Objective To assess the impact of maximal fat oxidation intensity(FATmax) exercise on arterial stiffness in overweight/obesity young men.Method Thirty two overweight/obesity young men (BMI≥25 kg/m2) without the habit of doing exercises,were divided into an exercise group (n=16) and a control group (n=16) randomly.The exercise group completed a 12-week walk/run(FATmax intensity,3-5 times/week,40-60 min/time) intervention,while the control group maintained normal life style without regular exercise.The body composition,cardiopulmonary function (VO2max),C reactive proteins (CRP) and blood hemodynamic were measured,and the brachial-ankle pulse wave velocity(baPWV) was observed to assess the arterial stiffness before and after the 12-week intervention.Results Significant decrease was observed in the average weight,body mass index,body fat mass,body fat percentage,baPWV,CRP,platelet/lymphocyte ratio (PLR) and neutrophil/lymphocyte ratio (NLR) (P＜0.05),while significant increase was found in the average VO2max and HDL-C(P＜0.05) after the training.After the intervention,the change in baPWV(△) was positively associated with △ CRP(r=0.604),△ NLR(r=0.535) and △ WBC (white blood count) (r=0.406) (P＜0.05,n=32),but negatively correlated with △ VO2max (r=-0.660,P＜0.05,n=32).Conclusion FATmax intensity aerobic exercise is effective to downregulate the inflammation,and improve aerobic fitness and vascular function.However,the potential mechanism of arterial stiffness improvement following exercise needs further study.

Aim To prepare evodiamine butyryl deriva-tive (EBD) and evodiamine butyryl derivative-loaded solid lipid nanoparticles (EBDLN), and study its re-lease in vitro,and to investigate its in situ gastrointesti-nal absorption. Methods EBD was prepared by a one-step synthetized method, and then EBDLN was prepared by a film dispersion method. Dynamic dialy-sis was used to evaluate drug release in vitro,and sin-gle-pass gastrointestinal perfusion was employed to study the gastrointestinal absorption of EDM,EBD and EBDLN. Results In identical release media, there were identical drug release tendencies of EBD and EB-DLN, but the release rate of EBDLN was faster than EBD. Compared with EDM and EBD, the Kavalues and Pappvalues of EBDLN in every perfusion segment increased significantly. The Kaof EBDLN in stomach, duodenum, jejunum, ileum and colon was 110.14-fold,56.70-fold,51.23-fold,45.70-fold and 127.23-fold of free EDM respectively. The Pappvalue of EB-DLN was 9.74-fold, 4.48-fold, 3.82-fold and 11.3-fold of that of free EDM. Conclusion EBDLN has sustained effect and can enhance the gastrointestinal absorption of EDM and EBD.

Alveolar epithelial type II (AT II) cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells (fibroblasts). Progenitor cells are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A (SPA) suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase (rAAV-SPA-TK) system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells (H441) transfected with TK gene had higher sensitivity to ganciclovir (GCV) than SPA low expression cells (A549). In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell (AT II) niche in vitro and in vivo.

Alveolar epithelial type II (AT II) cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells (fibroblasts). Progenitor cells are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A (SPA) suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase (rAAV-SPA-TK) system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells (H441) transfected with TK gene had higher sensitivity to ganciclovir (GCV) than SPA low expression cells (A549). In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell (AT II) niche in vitro and in vivo.
Antiviral Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cells, Cultured ; Dependovirus ; genetics ; Dose-Response Relationship, Drug ; Electrophoresis, Polyacrylamide Gel ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Ganciclovir ; pharmacology ; Gene Expression Regulation, Neoplastic ; Genes, Transgenic, Suicide ; genetics ; Genetic Vectors ; genetics ; Humans ; In Situ Nick-End Labeling ; Luciferases ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics ; Pulmonary Alveoli ; cytology ; metabolism ; Pulmonary Surfactant-Associated Protein A ; genetics ; metabolism ; Thymidine Kinase ; genetics ; metabolism

PURPOSE: Lapatinib is a candidate drug for treatment of trastuzumab-resistant, human epidermal growth factor receptor 2 (HER2)–positive gastric cancer (GC). Unfortunately, lapatinib resistance renders this drug ineffective. The present study investigated the implication of forkhead box O1 (FOXO1) signaling in the acquired lapatinib resistance in HER2-positive GC cells. MATERIALS AND METHODS: Lapatinib-resistant GC cell lines (SNU-216 LR2-8) were generated in vitro by chronic exposure of lapatinib-sensitive, HER2-positive SNU-216 cells to lapatinib. SNU-216 LR cells with FOXO1 overexpression were generated by stable transfection of a constitutively active FOXO1 mutant (FOXO1A3). HER2 and MET in SNU-216 LR cells were downregulated using RNA interference. The sensitivity of GC cells to lapatinib and/or cisplatin was determined by crystal violet assay. In addition, Western blot analysis, luciferase reporter assay and reverse transcription–polymerase chain reaction were performed. RESULTS: SNU-216 LR cells showed upregulations of HER2 and MET, but downregulation of FOXO1 compared to parental SNU-216 cells. FOXO1 overexpression in SNU-216 LR cells significantly suppressed resistance to lapatinib and/or cisplatin. In addition, FOXO1 negatively controlled HER2 and MET at the transcriptional level and was negatively controlled by these molecules at the post-transcriptional level. A positive crosstalk was shown between HER2 and MET, each of which increased resistance to lapatinib and/or cisplatin. CONCLUSION: FOXO1 serves as an important linker between HER2 and MET signaling pathways through negative crosstalks and is a key regulator of the acquired lapatinib resistance in HER2-positive GC cells. These findings provide a rationale for establishing a novel treatment strategy to overcome lapatinib resistance in a subtype of GC patients.
Blotting, Western ; Cell Line ; Cisplatin ; Down-Regulation ; Drug Resistance ; Gentian Violet ; Humans ; In Vitro Techniques ; Luciferases ; Parents ; Receptor, Epidermal Growth Factor ; Receptor, ErbB-2 ; RNA Interference ; Stomach Neoplasms ; Transfection ; Up-Regulation

OBJECTIVE: To investigate the effect and molecular mechanism of lncRNA X-inactive specific transcript (XIST) on the proliferation and apoptosis of acute myeloid leukemia cells KG1a. METHODS: Forty-one patients with acute myeloid leukemia from January 2017 to May 2019 treated in Beijing Aerospace Center Hospital were collected, as well as 20 patients who conformed to the international standard of iron deficiency anemia as control group. KG1a cells were divided into pcDNA group, pcDNA-XIST group, pcDNA-XIST+miR-NC group, and pcDNA-XIST+miR-196b group. Real-time fluorescence quantitative PCR was used to detect the expressions of XIST and miR-196b, CCK-8 was used to detect cell activity, flow cytometry was used to detect cell cycle and apoptosis, Western blot method was used to detect the protein expressions of cleaved-caspase3, pro-caspase3, Bax, and Bcl-2, and dual luciferase report experiment was used to detect the targeting relationship between XIST and miR-196b. RESULTS: The expression level of lncRNA XIST in bone marrow cells in the AML group was significantly lower than that in the iron deficiency anemia group (P<0.001). Compared with pcDNA group, the expression level of lncRNA XIST, proportion of cells in G₀/G₁ phase, apoptosis rate, and the expression levels of cleaved-caspase3 and Bax in the pcDNA-XIST group of KG1a cells were significantly increased (all P<0.001), while the expression level of miR-196b, cell viability, the proportion of S-phase cells, and the expression levels of pro-caspase3 and Bcl-2 were significantly decreased (all P<0.001). Compared with pcDNA-XIST group, the cell activity, proportion of S-phase cells, and the expression levels of pro-caspase3 and Bcl-2 in the pcDNA-XIST+miR-196b group were significantly increased (all P<0.001), while the proportion of cells in the G₀/G₁ phase, apoptosis rate, and the expression levels of cleaved-caspase3 and Bax decreased (all P<0.001). CONCLUSION Overexpression of lncRNA XIST can inhibit the proliferation of acute myeloid leukemia cells KG1a and promote apoptosis by down-regulating the expression of miR-196b.
Anemia ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Leukemia, Myeloid, Acute/metabolism* ; MicroRNAs/metabolism* ; Proto-Oncogene Proteins c-bcl-2 ; RNA, Long Noncoding/genetics* ; Sincalide ; bcl-2-Associated X Protein