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

OBJECTIVETo find out the mechanisms of redifferentiation and reversion of malignant human gastric cancer cells induced by ascorbic acid.

METHODSHuman gastric cancer cells grown in the laboratory were used. The Trypan blue dye exclusion method was used to determine the cell doubling time. The electrophoresis rate and colonogenic potential were the indices used to measure the rate of redifferentiation. The content of malondialdehyde (MDA) was measured using the thiobarbituric acid (TBA) method. The activities of superoxide dismutase (SOD), catalase (CAT) and the content of H202 were evaluated by spectrophotography.

RESULTSSix mmol/L ascorbic acid was used as a positive control. Human gastric cancer cells were treated with 75 microm hydrogen peroxide, which alleviated many of the malignant characteristics. For example, the cell surface charge obviously decreased and the electrophoresis rate dropped from 2.21 to 1.10 microm x s(-1) x V(-1) x cm(-1). The colonogenic potential, a measure of cell differentiation, decreased 90.2%. After treatment with ascorbic acid, there was a concentration- and time-dependent increase in hydrogen peroxide (H202) and the activity of superoxide dismutase (SOD). However, the activity of catalase (CAT) resulted in a concentration- and time-dependent decrease. SOD and 3-amino-1,2,4-triazole (AT) exhibited some effects, but there were statistically significant differences between the SOD and AT group and the H202 group.

CONCLUSIONSAscorbic acid induces growth inhibition and redifferentiation of human gastric cancer cells through the production of hydrogen peroxide.

Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Cell Differentiation ; drug effects ; Humans ; Hydrogen Peroxide ; metabolism ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; Tumor Cells, Cultured

Objective To systemically evaluate the effect of protective analgesia on preventing phantom limb pain(PLP)after amputa-tion.Methods Published articles from the earliest date available to June,2017 were recalled from Cochrane Library,PubMed,Embase,Web of Science,OVID,and Science Direct to collect prospective studies using protective analgesia in perioperative period to prevent PLP after amputation.Two reviewers screened literatures referring to studies according to the inclusion and exclusion criteria,and assessed the quality of them.Data of general information and incidence of PLP in the follow-up period were extracted and analyzed with RevMan 5.3 software. Results Six studies were included with a total of 256 patients in the one-month follow-up period including 127 cases in the protective anal-gesia group(group P)and 129 cases in the control group(group C),a total of 232 patients in the six-month follow-up period including 114 cases in group P and 118 cases in group C,and a total of 118 patients in the twelve-month follow-up period including 58 cases in group P and 60 cases in group C.The incidence of PLP were lower in group P than those in group C in the one-month follow-up period(RD=-0.21, 95%CI[-0.38,-0.04],Z=2.47,P=0.01)and in the six-month follow-up period(RD=-0.28,95%CI[-0.52,-0.05],Z=2.37,P=0.02),and it was not significant in the twelve-month follow-up period(RD=-0.20,95%CI[-0.48,0.09],Z=1.35,P=0.18).Conclusion Protective analge-sia in perioperative period can prevent against PLP after amputation in the recent time,however,it needs further observation in long-term.

BACKGROUNDAtrial AutoCapture™ (ACap™) was a new technological development that confirmed atrial capture by analyzing evoked response (ER) with a new method - paced depolarization integral ER detection - and optimized energy output to changes in the stimulation threshold. The purpose of this study was to evaluate the clinical performance of ACap™ function.

METHODSThis was a prospective, observational, nonrandomized two-center study. Between November 2008 and August 2014, 102 patients were enrolled from two different institutions. Data were collected by case report forms at enrollment, hospital discharge, and in-office follow-ups scheduled at 1, 2, 3, 6, and 12 months postimplantation.

RESULTSAmbulatory ACap™ function started to become available for 20.6% of patients at 1 day, then progressed to 30.4% at 7 days, 38.6% at 1 month, 41.6% at 2 months, 47.5% at 3 months, 53.5% at 6 months, and 63.4% at 1 year. The cause of the unsuccessful attempts to perform ACap™ threshold was ER/polarization <2:1. Availability for SD, BND, and HOCM indications had shown better results than AVB indication. For SD indication cases, feasibility was significantly better for SD with paroxysmal atrial fibrillation (pAF) than SD without pAF (78.4% vs. 35.0% at 1 year, n = 71, P< 0.001). At each stage of the clinical follow-ups, there had been a strict correlation between ACap™ measurements and those conducted manually with P 0.001 (n = 299).

CONCLUSIONSIt has been concluded that ACap™ function was safe and effective to confirm atrial threshold and reduce energy output automatically. ACap™ function is unavailable for some patients at early stages of the implantation; however, availability has been progressively increasing during follow-up.

OBJECTIVE: To reveal the effect of foods with different natures on cold or hot syndrome and gastrointestinal bacterial community structure in mice. METHODS: Forty-five 6-week-old male ICR Kunming mice of clean grade were divided into 5 groups, 9 per group, including the control (CK), hot nature herbs (HM), Hong Qu glutinous rice wine (RW), tea rice wine (TW), and cold nature herbs (CM) groups. Distilled water or corresponding herbs were administered to mice (0.01 mL/g body weight) in the 5 groups by gastric infusion respectively, once daily for 28 d. Appearance, behavior, and serum biochemical indicators, including 5-hydroxytryptamine (5-HT), thyroid stimulating hormone (TSH), noradrenaline (NE), cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), the hot nature index, as well as the gastrointestinal bacterial community structure were analyzed in all groups after treatment. RESULTS After supplementation for 28 d, CM and TW mice showed different degrees of cold syndrome, and HM and RW mice showed different degrees of hot syndrome. Compared with the HM and RW mice, the TSH, NE, cAMP levels and hot nature indices in the CM and TW mice were significantly decreased and 5-HT and cGMP levels were significantly increased (P<0.05). There was no obvious change in appearance or behavior in CK mice. Results of clustering analysis showed that the gastrointestinal bacterial community structures were highly similar in TW and CM mice as well as in RW and HM mice, and that they were from the same branch, respectively, when the distance was 0.02. The key microbes associated with cold syndrome were Lachnospiraceae uncultured, Lactococcus, etc., and the key microbes associated with hot syndrome were S24-7 norank, Ruminococcaceae uncultured, etc. CONCLUSION: The interventions with different nature foods could change cold or hot syndrome in mice, leading to changes in gastrointestinal bacterial community structure.

Intestinal organoids, also named "mini-guts", reconstitute sophisticated three-dimensional architecture recapitulating diversified intestinal epithelial cell types and physiology, which is driven by the proliferative and self-assembling characteristics of crypt stem cells. The initiation of organoids study relies on the identification of Lgr5+ crypt stem cells from different intestinal segments and the key role of EGF, Wnt, BMP/TGF-β, Notch signal pathways within the microenvironment during the cultivation process. Besides constituting polarized crypt-villus structures, these "mini-guts" exhibit various effective functions of intestinal epithelium. Since 2009 when the culture system of small intestinal organoids was established by Sato et al, intestinal organoids excel conventional intestinal models depending on genetical mutation in multiple aspects and thus have become the hotspot among the research on intestinal diseases. Combined with genomics, material science and engineering, "mini-guts" have been widely applied to the research on intestinal development, intestinal transport physiology, epithelial barrier, pathogen-host interaction and the study on cystic fibrosis, infectious diarrhea, ulcerative colitis, Crohn's disease, intestinal cancer, etc. In this review, we summarize the new insights introduced by organoid into the research on intestinal diseases, and related research advances and applications.
Humans ; Intestinal Mucosa ; Intestinal Neoplasms ; Intestines ; Organoids ; Stem Cells ; Tumor Microenvironment

Background:Renal cell carcinoma (RCC) is frequently associated with paraneoplastic inflammatory syndrome (PIS). This study aimed at exploring the connections between the survival rate and specific gene alterations and the potential mechanism.

Methods:We retrospectively studied 69 surgical RCC cases from August 2014 to February 2016, including 18 cases of clear cell RCC (ccRCC) demonstrating elevated pretreatment serum C-reactive protein (CRP, Group A). Twelve of the 18 cases were symptomized with febrile episode. We also selected 49 cases of ccRCC with normal pretreatment CRP (Group B). Using 22 microsatellite markers, we compared the incidence of loss of heterozygosity (LOH) between Group A and Group B. All statistical tests are two-sided.

Results:The 3p LOH was common in both Group A (89%) and Group B (92%). The frequency of 14q LOH in Group A (16 of 18) was higher than Group B (4 of 49, χ= 40.97 P < 0.0001). The 3p and 14q LOH were the characteristics of ccRCC with elevated acute phase reactants, including PIS, regardless of the presence of metastasis. On the contrary, 14q LOH was a rare genomic alternation in advanced-staged ccRCC without PIS. The overall survival of patients with elevated CRP (33.3%) was lower than its counterparts (6.1%, hazard ratio=1.852, P < 0.0001) in Kaplan-Meier curve.

Conclusions:The results imply that the disruption of a 14q gene(s) might result in not only the inflammatory manifestations in the tumor host but also the poor survival rate as well. The isolation of the gene(s) on 14q might be a vital goal in the treatment of PIS-associated RCC.

: Objective: To investigate the efficacy of fistula stent made by 3D printing technique in the treatment of enteroatmospheric fistula. Methods: A descriptive case series study was carried out. INCLUSION CRITERIA: (1) patients with open abdomen; (2) patients with enteroatmospheric fistula. EXCLUSION CRITERIA: (1) patient with two or more fistulas; (2) distal obstruction; (3) bowel stenosis over 50%. According to above criteria, 17 EAF patients admitted to the General Surgery Department of Jinling Hospital from June 2019 to January 2020 were retrospectively included in study. Based on the intestinal radiography, CT reconstruction and finger exploration, the size of fistula, the diameter of the intestinal tube and the angle of the intestinal lumen around the fistula were assessed. The 3D printing fistula stent was designed and established based on estimated data, and then placed through the fistula. OUTCOME MEASUREMENTS (1) success rate of stent implantation; (2) outflow of intestinal contents after implantation; (3) tolerated exercise time; (4) receiving definite operation time for intestinal fistula; (5) time to recovery of enteral nutrition. The t-test was used to compare the outflow amount of intestinal content before and after the stent implantation and the tolerated exercise time. The changes of the outflow amount of intestinal content and tolerated exercise time were analyzed by repeated measurement ANOVA. Results: Seventeen EAF patients with open abdomen included 13 males and 4 females. All the patients successfully received intestinal fistula stent implantation. Gastrointestinal angiography 2 days after implantation showed that the digestive tract was unobstructed, and the stent was successfully kept in place until definite surgery. No stent implantation-related adverse reactions were found in patients undergoing definite intestinal fistula surgery. The average outflow amount of intestinal fluid within 7 days after implantation decreased from (702.7±198.9) ml/d to on the first day after implantation (45.8±22.4) ml/d on the 7th day(F=10.380, P<0.001). The ambulatory time and exercise time of patients continued to increase after stent implantation. The average tolerated exercise time within 14 days after stent implantation increased from (9.1±3.8) min/d to (106.9±21.8) min/d (F=41.727, P<0.001). Within 120 days after stent implantation, 15 patients successfully underwent definite surgery for intestinal fistula and reconstruction of abdominal wall. Patients needed a median (IQR) of 3 (2, 5) days to recover enteral nutrition. The average time from stent placement to surgery was (87.2±17.6) days. Two patients died of severe abdominal infection with multiple organ failure. Conclusion: 3D printing fistula stent can significantly and the outflow of intestinal contents and the difficulty of nursing, and help to restore enteral nutrition and rehabilitation exercise as soon as possible in EAF patients with open abdomen.
Abdominal Wall ; Female ; Humans ; Intestinal Fistula/surgery* ; Male ; Printing, Three-Dimensional ; Retrospective Studies ; Stents ; Treatment Outcome

Itch is an unpleasant sensation that provokes the desire to scratch. While acute itch serves as a protective system to warn the body of external irritating agents, chronic itch is a debilitating but poorly-treated clinical disease leading to repetitive scratching and skin lesions. However, the neural mechanisms underlying the pathophysiology of chronic itch remain mysterious. Here, we identified a cell type-dependent role of the anterior cingulate cortex (ACC) in controlling chronic itch-related excessive scratching behaviors in mice. Moreover, we delineated a neural circuit originating from excitatory neurons of the ACC to the ventral tegmental area (VTA) that was critically involved in chronic itch. Furthermore, we demonstrate that the ACC→VTA circuit also selectively modulated histaminergic acute itch. Finally, the ACC neurons were shown to predominantly innervate the non-dopaminergic neurons of the VTA. Taken together, our findings uncover a cortex-midbrain circuit for chronic itch-evoked scratching behaviors and shed novel insights on therapeutic intervention.
Mice ; Animals ; Gyrus Cinguli/physiology* ; Pruritus/pathology* ; Mesencephalon ; Cerebral Cortex/pathology* ; Neurons/pathology*

BACKGROUND: Prospective analyses have yet to identify a consistent relationship between sleep duration and the incidence of gastrointestinal (GI) cancers. The effect of changes in sleep duration on GI cancer incidence has scarcely been studied. Therefore, we aimed to examine the association between baseline sleep duration and annual changes in sleep duration and GI cancer risk in a large population-based cohort study. METHODS: A total of 123,495 participants with baseline information and 83,511 participants with annual changes in sleep duration information were prospectively observed from 2006 to 2015 for cancer incidence. Cox proportional-hazards models were used to calculate hazard ratios (HRs) and their confidence intervals (CIs) for GI cancers according to sleep duration and annual changes in sleep duration. RESULTS: In baseline sleep duration analyses, short sleep duration (≤5 h) was significantly associated with a lower risk of GI cancer in females (HR: 0.31, 95% CI: 0.10-0.90), and a linear relationship between baseline sleep duration and GI cancer was observed (P = 0.010), especially in males and in the >50-year-old group. In the annual changes in sleep duration analyses, with stable category (0 to -15 min/year) as the control group, decreased sleep duration (≤-15 min/year) was significantly associated with the development of GI cancer (HR: 1.29; 95% CI: 1.04-1.61), especially in the >50-year-old group (HR: 1.32; 95% CI: 1.01-1.71), and increased sleep duration (>0 min/year) was significantly associated with GI cancer in females (HR: 2.89; 95% CI: 1.14-7.30). CONCLUSIONS Both sleep duration and annual changes in sleep duration were associated with the incidence of GI cancer.
Cohort Studies ; Female ; Gastrointestinal Neoplasms/etiology* ; Humans ; Incidence ; Male ; Middle Aged ; Proportional Hazards Models ; Prospective Studies ; Risk Factors ; Sleep