Animals ; CCAAT-Enhancer-Binding Proteins ; analysis ; chemistry ; physiology ; Gene Expression Regulation ; Humans ; Liver ; chemistry ; metabolism ; Liver Diseases ; etiology ; metabolism

OBJECTIVETo study the effect of RAR-beta transfection plus treatment with the corresponding ligand ATRA on the proliferation and phenotype of platelet-derived growth factor (PDGF)-activated hepatic stellate cells (HSC).

METHODSPDGF-activated hepatic stellate cells of rats were transfected with eukaryotic expression vector pCMV-script-RAR-beta, which was verified by western blot. The proliferation of transfected HSC was assayed by BrdU incorporation as well as MTT methods. Their phenotype (alpha-SMA and desmin) was observed by immunocytochemistry assay with image analysis and RAR-beta protein expression was detected by western blot.

RESULTSTransfection of RAR-beta gene and treatment with ligand ATRA could increase the expression of RAR-beta protein for at least 144h and inhibit the proliferation and the expression of alpha-SMA and desmin in PDGF-activated HSC. Significant statistical differences were perceived comparing with sham-transfected, only-PDGF treated, non-ligand treated and irrelevant ligand-treated HSC.

CONCLUSIONSTransfected with RAR-beta gene as well as using related ligand ATRA could suppress the proliferation and reverse the activation phenotype of activated HSC.

Animals ; Blotting, Western ; Cell Division ; Liver ; cytology ; Phenotype ; Platelet-Derived Growth Factor ; pharmacology ; Rats ; Receptors, Retinoic Acid ; physiology ; Transfection ; Tretinoin ; pharmacology

OBJECTIVEThe expression of C/EBPalpha protein and mRNA during automatically activation process in primary cultures of HSCs were observed in order to explore its possible association with the proliferation and activation of HSCs.

METHODSImmunocytochemistry, Western blot and RT-PCR were used to evaluated the expression of C/EBPalpha protein and mRNA; as well as the expression of alpha-SMA, Desmin, MMP2, type I procollagen (alpha1). The eukaryotic vector harboring the full length cDNA of C/EBPalpha was transfected into activated HSC, then immunocytochemistry was applied to confirm the transfection and evaluate the effect of transfection on the proliferation of HSC by calculating the PCNA-positive cells. The morphological changes of HSC were observed by use of phase-contrast microscope.

RESULTSConstitutive expression of mRNA and protein of C/EBPalpha were detected in primarily cultured HSCs, and the protein was seen in both nuclei and cytoplasm with the latter being dominant. Their expression levels reached highest at day 2 of the culture, then decreased gradually when continually cultured to the day 4, 7, 10, on the other hand, the expression of alpha-SMA, MMP2 and ColI(alpha1) increased steadily. Transient transfection was verified by the fact that much more and stronger C/EBPalpha stain was observed in transfected HSCs than in void-vector transfected cells. In C/EBPalpha gene transfected HSCs, the number of PCNA-positive cells dramatically decreased compared with the void-vector transfected cells 24h after transfection. In addition, the C/EBPalpha gene transfected HSCs died 36 h after transfection, a few surviving cells became longer and thinner in morphology, however the void-vector transfected cells almost all remained alive.

CONCLUSIONSC/EBPalpha was likely involved in the HSCs activation, and over-expressed C/EBPalpha by transfection had inhibitory influence on the proliferation of cultured rat HSCs.

Animals ; CCAAT-Enhancer-Binding Protein-alpha ; genetics ; Cells, Cultured ; Collagen Type I ; genetics ; Liver ; cytology ; metabolism ; Male ; Matrix Metalloproteinase 2 ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo investigate the expression of phosphatase of regeneration liver-3(PRL-3) protein and its relationship with tumor invasion and metastasis in human colorectal carcinoma,and elucidate prognostic value.

METHODSImmunohistochemistry method was applied to detect the PRL-3 expression in the primary tumor specimens and paired paratumor normal tissues from 46 colorectal carcinoma patients, the adenoma tissues from 6 patients with colorectal adenoma, all the metastatic lymph nodes from 29 cases and the metastatic liver lesions from 6 cases. The relationship between PRL-3 expression and clinicopathologic parameters was analyzed and a survival curve was achieved according to Kaplan-Meier method.

RESULTSNo or weak PRL-3 protein expression was detected in normal colorectal mucosa and colorectal adenoma. In colorectal carcinoma tissues, PRL-3 expression was confirmed in 26 of 46 cases (56.5%) of primary colorectal carcinomas (with lymph node metastasis 63.0%, without lymph node metastasis 37.0%, P=0.001), 26 of 29 (89.7%) lymph node metastases, and 5 of 6 liver metastases. The expression of PRL-3 was assembled in the cytoplasm of carcinoma cells and more intensively on the cell membrane.Analysis of the relationship between PRL-3 expression and the clinicopathologic features showed that PRL-3 expression was closely associated with tumor stage (P=0.019), lymph node metastasis (P=0.026), but no relationship with age, sex, tumor size, degree of differentiation was founded (P<0.05). The mean follow-up time was 41.4 months and results showed that patients with positive expression of PRL-3 had a significantly poorer prognosis than those with negative PRL-3 expression group(P=0.032).

CONCLUSIONSPRL-3 protein plays a novel role in tumor progression and metastasis of colorectal carcinoma. PRL-3 can be expected to be a potential predictive biomarker for identifying the prognosis in colorectal carcinoma patients.

Adult ; Aged ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Liver Neoplasms ; metabolism ; secondary ; Liver Regeneration ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Neoplasm Staging ; Prognosis ; Protein Tyrosine Phosphatases ; metabolism

Abstract: Objective To investigate the epidemiological characteristics of pathogens causing bloodstream infection in hematology patients during treatment and to compare the effects of allogeneic hematopoietic stem cell transplantation (HSCT) on them, so as to provide evidence for the diagnosis and treatment of bloodstream infection. Methods A total of 292 cases with bloodstream infection in hematology wards of the PLA General Hospital were collected from 2017 to 2021, which were divided into HSCT group and N-HSCT group according to whether performed HSCT or not. The epidemiological characteristics and influence of pathogenic bacteria in blood stream infection were analyzed and compared between the two groups. Results A total of 362 strains of pathogenic bacteria were collected from 292 cases, including 106 strains in HSCT group (84 cases) and 256 strains in N-HSCT group (208 cases). Bloodstream infections were more common in acute myeloid leukemia (130/392, 44.52%), followed by non-Hodgkin's lymphoma (74/292, 25.34%). The rate of once bloodstream infection in HSCT group was higher than that in N-HSCT Group, but the rate of twice bloodstream infections in N-HSCT group was higher. Gram-negative Bacilli were the most common pathogens (56.08%), with Escherichia coli being absolutely dominant (109/362, 30.11%), followed by Klebsiella pneumoniae (39/362, 10.77%). Coagulase-negative staphylococci (CoNS) (107/362, 29.56%) were the most common Gram-positive cocci. The detection rate of fungi in HSCT group (10/106, 9.43%) was significantly higher than that in N-HSCT Group (3.52%). The drug resistance rate of the common pathogenic bacteria was at a high level, and there was a certain proportion of multi-drug resistant strains (except for Pseudomonas aeruginosa). The resistance rates of CoNS to penicillin, gentamicin, moxifloxacin, clindamycin and rifampicin in HSCT group were higher than those in N-HSCT Group. The resistance rate of Escherichia coli to piperacillin/tazobactam, cephalosporins and etapenem in HSCT group was significantly higher than that in N-HSCT group. Conclusions The pathogens of blood stream infection in hematology patients are complicated and various. It is difficult for clinical diagnosis and treatment to detect multiple infections and multiple pathogens. HSCT patients have a higher risk of fungal bloodstream infection and more multi-drug resistant strains detected. Therefore, the identification of bloodstream infection and multi-drug resistant strains associated with HSCT patients should prompt surveillance.

OBJECTIVETo explore the effects of surgical procedure combined with the intravascular minimal invasive technique for the treatment of deep vein thrombosis (DVT) of lower extremity.

METHODSAt the curse of disease from six hours to ninety days, one hundred and two patients with DVT including one hundred and three lower extremities had received surgical procedure and intravascular minimal invasive treatment.

RESULTSThere were not procedure-related morbidities in 102 cases, and symptoms disappeared, all procedures were successful based on angiography. The detecting head for the intravascular ultrasound ablation was entered to inferior vena cava (IVC) in 74 cases (78%), Forgarty catheter was entered to IVC in 21 cases (21%), the stenosis in the confluence of the common iliac vein and IVC was dilated by sacculus rotundus catheter in 89 cases (88%), including 9 patients underwent percutaneous transluminal stenting. One hundred and two patients followed up for twenty months, follow-up by angiography showed no restenosis in 91 cases, restenosis in ilio-femoral vein in 1 cases, and thrombus recontouring in 4 cases, as well as 6 cases died caused by primary disease.

CONCLUSIONSurgical procedure combined with the intravascular minimal invasive technique is a safe and effective therapeutic method for DVT.

Adult ; Aged ; Aged, 80 and over ; Angioplasty, Balloon ; Catheter Ablation ; methods ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Lower Extremity ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Stents ; Ultrasonic Therapy ; Venous Thrombosis ; surgery ; therapy

OBJECTIVETo evaluate the effects of antiarrhythmic peptide (AAP10) on ventricular arrhythmias in rabbits with healed myocardial infarction (OMI).

METHODSThirty rabbits were randomly divided into three groups (n = 10 each): Sham group, left thoracotomy was performed without coronary ligation; OMI group and OMI + AAP10 group, the circumflex coronaries were ligated. Three months post operation, the electrophysiological and antiarrhythmic effects of AAP10 were assessed in the arterially perfused rabbit left ventricular wedge preparation. Sham and OMI group were perfused with Tyrode's solution and OMI + AAP10 group was perfused with Tyrode's solution + AAP10 (80 nmol/L). Transmembrane action potentials were recorded simultaneously from endocardium and epicardium together with a transmural ECG by use of 2 separate intracellular floating microelectrodes. The stimulus-response-interval (SRI) of the epicardium and the incidence of ventricular tachycardia (VT) were observed. Whole heart and left ventricular weights, the left ventricular thickness at infarct border zone were measured.

RESULTSWhole heart and left ventricular weights as well as the left ventricular thickness at the infarct border zone significantly increased post infarction. VT was induced in 8 out of 10 rabbits in OMI group and in 2 out of 10 rabbits in OMI + AAP10 group (P < 0.05). SRI was also significantly shortened in OMI + AAP10 group compared to OMI group [SRI-1: (20.59 +/- 0.79) ms vs. (28.71 +/- 0.55) ms; SRI-2: (30.42 +/- 0.74) ms vs. (38.67 +/- 0.49) ms, all P < 0.01]. However, the action potential morphology and duration were similar between OMI and OMI + AAP10 groups.

CONCLUSIONThe antiarrhythmic peptide (AAP10) can increase gap junctional intercellular conductance without affecting the action potential morphology and duration and decrease the incidence of inducible ventricular tachycardia.

Animals ; Arrhythmias, Cardiac ; etiology ; prevention & control ; Male ; Myocardial Infarction ; complications ; physiopathology ; Oligopeptides ; pharmacology ; Rabbits ; Random Allocation

AIM: To explore the neuroprotective effect of fasudil combined with bone marrow -derived neural stem cells ( BM-NSCs) on the mice with experimental autoimmune encephalomyelitis ( EAE).METHODS: Female C57BL/6 mice (8~10 weeks old, n=32) were immunized with myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) to establish chronic EAE model .The mice were randomly divided into control ( ddH2 O ) group, fasudil group , BM-NSCs group , and fasudil+BM-NSCs group .The clinical score and body weight were recorded every other day .The expression of neurotrophic factors was determined by immunofluorescence staining .RESULTS:In comparison with ddH2O group, fasud-il combined with BM-NSCs delayed onset and ameliorated severity of EAE .The numbers of brain-derived neurotrophic fac-tor, glial cell-derived neurotrophic factor , nerve growth factor , neurotrophin-3 and ciliary neurotrophic factor positive cells in fasudil group, BM-NSCs group and fasudil +BM-NSCs group were all increased in various extents .In particularly, the expression of these neurotrophic factors in fasudil +BM-NSCs group was significantly higher than that in the mice treated with fasudil or BM-NSCs alone (P<0.01).CONCLUSION:Fasudil combined with BM-NSCs promotes the expression of neurotrophic factors and improves microenvironment of central nervous system , thus playing a positive role in neural restora-tion and regeneration through a synergistic and superimposed effect .

OBJECTIVETo evaluate the effects of rapamycin (RPM)-loaded poly (lactic-co- glycolic) acid (PLGA) nanoparticles (NPs) on the proliferation, distribution of cell cycle, and expression of p27 protein in human umbilical arterial vascular smooth muscle cell (HUASMC) in vitro.

METHODSThe primarily culture model of HUASMC was successfully established by explant-attached method in vitro. The cells were administrated with different doses of RPM, and RPM-PLGA NPs were observed as treat groups compared with PLGA NPs and M231-SMGs medium cultured group. The effect of RPM-PLGA NPs on proliferation of HUASMC was assessed using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) colorimetry method. The influences of RPM-PLGA NPs on the cell cycle and cellular growth kinetics of HUASMCs were tested by flow cytometry. The effect of RPM-PLGA NPs on the expression of p27 protein of HUASMCs was assessed through an immunohistochemical method.

RESULTSCompared with the control group, the proliferation of HUASMCs was inhibited by 50 microg/L and higher concentration of RPM-PLGA NPs in a dose-dependent manner (P < 0.05). The numbers of cells entering cell cycle of S/G2/M phases were significantly lower in RPM-PLGA NPs and RPM treated groups. Histologically, the expression of p27 were up-regulated in 500 microg/L RPM-PLGA NPs and 100 microg/L RPM treated group (all P < 0.01 ) when compared with the control group.

CONCLUSIONSRPM-PLGA NPs has a similar effects as RPM in inhibiting the growth of in vitro cultured HUASMC. It can remarkably suppress the expression of in vitro cultured HUASMC p27 protein, arrest its cell cycle at G1/S phase, and inhibit its proliferation.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Drug Carriers ; Humans ; Lactic Acid ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Nanoparticles ; Polyglycolic Acid ; Sirolimus ; administration & dosage ; pharmacology ; Umbilical Arteries ; cytology

OBJECTIVETo explore whether delayed umbilical cord clamp timing of newborn can improve iron stores of infant period and growth and development.

METHODSMother-infant pairs were randomly assigned to early clamping (94 cases, < 15 s after delivery) and delayed clamping (64 cases, 1 min after delivery) by draw lots, and followed up until 4 months postpartum. Infant hematological status, iron status, the level of growth and development of infants after 4 months were measured respectively. Transcutaneous bilirubin at the third day after delivery was also measured.

RESULTSAt 4 month age, the median of serum ferritin and mean of corpuscular volume value in delayed group were 87.30 µg/L and (79.62 ± 4.13) fl, significantly higher than the values in early group (64.3 µg/L, (78.21 ± 4.38) fl), respectively (Z = -2.36, t = 2.23, both P values < 0.05). The hematocrit value was (33.59 ± 2.48)%, higher than that in early group (32.76 ± 2.69)% (t = 1.95, P = 0.05). There was no statistically significant difference at other iron nutrition indicators and infants' weight and body length at 4 month (P > 0.05). Under the different cut-off values (hemoglobin (Hb) < 105 g/L and Hb < 110 g/L, respectively), the prevalence of anemia in delay and early clamping group were 6.25% (4/64), 21.86% (14/64), and 12.77% (12/94), 34.04% (32/94), respectively (both P values > 0.05).

CONCLUSIONDelayed umbilical cord clamp timing until 1 min can improve iron stores of breastfed infants at 4 month; there is no significant adverse effects to growth.

Adult ; Breast Feeding ; Child Development ; Female ; Humans ; Infant ; Iron, Dietary ; Male ; Nutritional Status ; Pregnancy ; Umbilical Cord ; Young Adult