Long Non-Coding RNA Expression Profiles for the Characterization of Different Bladder Cancer Grade.

BACKGROUND/AIMS: Bladder cancer (BC) is one of the most frequent urologic tumors worldwide. However, long non-coding RNA(lncRNA) expression profiles in BC progression remain unclear. This study aimed to explore lncRNA expression profiles in different grades of bladder cancer and normal urothelium tissues. METHODS: We performed high-throughput sequencing in BC tissues of different grade and obtained the expression profiles of its lncRNAs. Then, aberrantly expressed lncRNAs were validated by quantitative reverse transcription polymerase chain reaction (RT-PCR). Gene Ontology (GO) and pathway analyses were used to investigate the potential function of these lncRNAs. Co-expresson network was constructed to explore the relationship between lncRNAs and target mRNAs. RESULTS: We identified 252 aberrantly expressed lncRNAs in high-grade BC while compared to low-grade BC, and 269 lncRNAs in high-grade BC while compared to normal urothelium. Notably, we found 33 overlapped lncRNAs. Subsequently, 7 lncRNAs were selected from the overlapped part and confirmed by RT-PCR. GO and pathway analyses showed that these dysregulated lncRNAs participated in cell migration, cell adhesion, as well as Ras signaling pathway. Co-expression network and The Cancer Genome Atlas (TCGA) data showed LUCAT1 and CCNB1 had positive relationship in regulating the progress of bladder cancer. CONCLUSION: Our findings revealed the significant role of lncRNAs in the development process of bladder cancer.

Effects of early enteral nutrition on T helper lymphocytes of surgical septic patients: A retrospective observational study.

The aim of this study was to investigate the effects of early enteral nutrition (EEN) on T helper lymphocytes and the subpopulations ratios of surgical septic patients.We performed a retrospective study including 107 eligible patients from February 2014 to December 2015. Patients were divided into EEN, delayed enteral nutrition (DEN), or total parenteral nutrition (TPN) group according to the duration before enteral feeding. Th1, Th2, Th17, and Treg lymphocyte percentages were collected on days 3, 7, and 14 after admission. The disease severity and clinical outcome variables were also recorded.The Th1, Th17 percentages, and Th1/Th2, Th17/Treg ratios of EEN group were significantly lower than those of DEN or TPN group on the 14th day after admission (P < .05). Compared with TPN, DEN might have a tendency to decrease the Th1 and Th17 percentages. EEN could improve the disease severity and clinical outcomes of septic patients, however, no difference on 28-day mortality was found between EEN and DEN group.EEN could improve the dysregulation of Th1/Th2 and Th17/Treg ratios during early stage of sepsis. Compared with DEN, EEN could improve the disease severity and clinical outcomes, but not decrease the 28-day mortality of surgical septic patients.

Risk factors and prognosis of hypoalbuminemia in surgical septic patients.

The aim of this study was to investigate the risk factors of hypoalbuminemia and effects of different albumin levels on the prognosis of surgical septic patients. We preformed a retrospective clinical study including 135 adult patients from September 2011 to June 2014. The albumin levels and severity markers were recorded during the first 48 h after enrollment, and logistic regression analyses were used to determine the risk factors. The outcomes of patients with different albumin levels were also compared. The acute physiology and chronic health evaluation II (APACHE II) score (OR 1.786, 95% CI [1.379-2.314], P < 0.001), C-reactive protein (CRP) (OR 1.016, 95% CI [1.005-1.027], P = 0.005), and blood lactate (OR 1.764, 95% CI [1.141-2.726], P = 0.011) were established as the independent risk factors of hypoalbuminemia in patients with surgical sepsis. The severity markers and outcomes of patients with albumin levels ≤20 g/L were significantly worse than that of 21-25 g/L and ≥26 g/L, whereas the latter two groups had similar prognosis. Every 1 g/L decrease of albumin level below the optimal cut-off (23 g/L) was associated with a 19.4% increase in hospital mortality and a 28.7% increase in the incidence of multiple organ dysfunction syndrome. In conclusion, APACHE II score (≥14.5), CRP (≥34.25 mg/L), and blood lactate (≥.35 mmol/L) were established as the independent risk factors of hypoalbuminemia in the early stage of surgical sepsis. Patients with baseline albumin level ≤20 g/L had worse prognosis than that of albumin level ≥21 g/L. Albumin levels were negatively correlated the prognosis of surgical sepsis when below about 23 g/L.

Effects of early enteral nutrition on immune function of severe acute pancreatitis patients.

AIM: To investigate the effects of early enteral nutrition (EEN) on the immune function and clinical outcome of patients with severe acute pancreatitis (SAP). METHODS: Patients were randomly allocated to receive EEN or delayed enteral nutrition (DEN). Enteral nutrition was started within 48 h after admission in EEN group, whereas from the 8(th) day in DEN group. All the immunologic parameters and C-reactive protein (CRP) levels were collected on days 1, 3, 7 and 14 after admission. The clinical outcome variables were also recorded. RESULTS: Sixty SAP patients were enrolled to this study. The CD4+ T-lymphocyte percentage, CD4+/CD8+ ratio, and the CRP levels in EEN group became significantly lower than in DEN group from the 7(th) day after admission. In contrast, the immunoglobulin G (IgG) levels and human leukocyte antigen-DR expression in EEN group became significantly higher than in DEN group from the 7(th) day after admission. No difference of CD8+ T-lymphocyte percentage, IgM and IgA levels was found between the two groups. The incidences of multiple organ dysfunction syndrome, systemic inflammatory response syndrome, and pancreatic infection as well as the duration of intensive care unit stay were significantly lower in EEN group than in DEN group. However, there was no difference of hospital mortality between the two groups. CONCLUSION: EEN moderates the excessive immune response during the early stage of SAP without leading to subsequent immunosuppression. EEN can improve the clinical outcome, but not decrease the hospital mortality of SAP patients.

TSLC1 expression discriminates cutaneous melanomas from dysplastic nevi.

Cutaneous melanoma is a malignant tumor of melanocytes that causes the majority of skin cancer-related deaths. However, sometimes, discrimination between dysplastic nevi and early melanomas is difficult, even for experienced pathologists. Besides histology, the silencing of tumor suppressor genes aids in the diagnosis of melanoma. We have shown previously that tumor suppressor in lung cancer 1 (TSLC1) is a tumor suppressor gene, and its silencing through aberrant promoter methylation is associated with the generation of cutaneous melanoma. To examine TSLC1 expression in melanocytic skin lesions to determine whether it can serve as a diagnostic marker in histologically questionable lesions. Cytoplasmic localization of the expression of the TSLC1 gene was detected by immunohistochemistry; the levels of TSLC1 mRNA and protein were detected by quantitative real-time reverse transcription-PCR and western blot, respectively. Using immunohistochemistry, the average TSLC1 expression levels in cutaneous melanomas decreased approximately 3.6-fold (n=20) as compared with dysplastic nevi (n=30) and 3.7-fold as compared with normal skin (n=25). The average expression levels of TSLC1 mRNA and protein in dysplastic nevi lesions and normal skin were significantly higher than the levels in cutaneous melanomas. No significant changes in TSLC1 mRNA and protein expressions were found between normal skin and dysplastic nevi. Our results show that a loss of TSLC1 frequently occurs in cutaneous melanoma, and indicate that it could serve as a diagnostic marker for cutaneous melanoma in histologically questionable lesions.

[Effect of acidic fibroblast growth factor on mitogen-activated protein kinase activity in small intestinal epithelium after ischemia/reperfusion injury in rat].

OBJECTIVE: To investigate the effect of acidic fibroblast growth factor (aFGF) on p44/p42 mitogen-activated protein kinase (p44/p42 MAPK, extracellular signal-regulated protein kinase, ERK1/2) activity in small intestinal epithelium in rat after ischemia/reperfusion (I/R) injury and its relation with the change in small intestinal epithelium in rat after I/R injury as well as the change in proliferation of epithelial cells. METHODS: Superior mesenteric artery (SMA) was occluded to produce ischemia of the intestine for 45 minutes followed by reperfusion to reproduce I/R injury. One hundred and thirty-two Wistar rats were randomly divided into sham-operation group, I/R group, aFGF treatment group (4 microg of aFGF was injected into jugular vein after reperfusion), and PD98059 (antagonist of ERK 1/2) pretreatment group (7.5 microg of PD98059 was injected via tail vein before ischemia and 4 microg of aFGF was injected via jugular vein after reperfusion). The activity of ERK1/2 and proliferation rate of small intestinal epithelial cells were determined before ischemia and 15 minutes, 30 minutes, 1 hour, 2 hours, 6 hours, 12 hours and 24 hours after reperfusion. RESULTS: The activity of ERK1/2 in small intestinal epithelium was higher in aFGF treatment group than in I/R control group, and the proliferation rate in small intestinal epithelial cells was significantly enhanced in aFGF treatment group. PD98059, the specific inhibitor of ERK1/2, inhibited ERK1/2 activity and reduced the proliferation rate of small intestinal epithelial cells in aFGF treatment group. CONCLUSION: aFGF can promote small intestinal epithelial cell proliferation in I/R injury rats, and this may be related to activation of ERK1/2 in small intestinal epithelium.

[Protective effects of acidic fibroblast growth factor on intestinal ischemia/reperfusion in rats].

OBJECTIVE: To observe the change in hepatic and renal functions, change in the plasma D-lactate level, and the expression of proliferating cell nuclear antigen (PCNA) after intestinal I/R injury, so as to explore the effects of reconstructive human acid fibroblast growth factor(aFGF) on intestinal I/R injury in rats. METHODS: One hundred and twenty-six Wistar rats were divided into sham-operated, ischemia (45 minutes) plus reperfusion, reconstructive human aFGF treatment (2, 4, 8 microg aFGF) and wild type aFGF(2, 6, 12, and 24 hours, respectively) groups. Hepatic and renal functions and the levels of plasma D-lactate were determined and the expression of PCNA was assessed. RESULTS: Compared with all other groups, bowel barrier function and hepatic and renal functions showed most marked deterioration in sham-operated group. The damages were less marked in reconstructive human aFGF group compared with other groups 24 hours after ischemia/reperfusion of the intestine, and the protective effect was best shown when 4 microg of aFGF was given. The trend of expression of PCNA was similar to that of changes in D-lactate level. CONCLUSION: Wild type reconstructive human aFGF treatment significantly improves the outcome of ischemia/reperfusion injury to the intestine, and the effect is dose-dependent.

Inhibition of p38 mitogen-activated protein kinase may decrease intestinal epithelial cell apoptosis and improve intestinal epithelial barrier function after ischemia- reperfusion injury.

AIM: To investigate the role of p38 mitogen-activated protein kinase in rat small intestine after ischemia-reperfusion (I/R) insult and the relationship between activation of p38 MAPK and apoptotic cell death of intestine. METHODS: Ninety Wistar rats were divided randomly into three groups, namely sham-operated group (C), I/R vehicle group (R) and SB203580 pre-treated group (S). In groups R and S, the superior mesenteric artery (SMA) was separated and occluded for 45 min, then released for reperfusion for 0.25, 0.5, 1, 2, 6, 12 and 24 h. In group C, SMA was separated without occlusion. Plasma D-lactate levels were examined and histological changes were observed under a light microscope. The activity of p38 MAPK was determined by Western immunoblotting and apoptotic cells were detected by the terminal deoxynucleotidyl transferase (TdT)-mediated dUDP-biotin nick end labeling (TUNEL). RESULTS: Intestinal ischemia followed by reperfusion activated p38 MAPK, and the maximal level of activation (7.3-fold vs sham-operated group) was reached 30 min after I/R. Treatment with SB 203580, a p38 MAPK inhibitor, reduced intestinal apoptosis (26.72+/-3.39% vs 62.50+/-3.08% in I/R vehicle, P<0.01) and decreased plasma D-lactate level (0.78+/-0.15 mmol/L in I/R vehicle vs 0.42+/-0.17 mmol/L in SB-treated group) and improved post-ischemic intestinal histological damage. CONCLUSION: p38 MAPK plays a crucial role in the signal transduction pathway mediating post-ischemic intestinal apoptosis, and inhibition of p38 MAPK may attenuate ischemia-reperfusion injury.

[Protective effects of modified recombinant human acidic fibroblast growth factor on small intestine after ischemia/reperfusion injury in rats].

OBJECTIVE: To explore the protective effect of modified recombinant human acidic fibroblast growth factor (rhaFGF) on small intestinal after ischemia/reperfusion (I/R) injury in rats. METHODS: The clamp on the superior mesenteric artery (SMA) was removed after clamping it for 45 minutes to replicate I/R injury of the intestine in the rat. Rats were then divided randomly into sham operation group, normal saline treatment group and rhaFGF treatment group, in which the rats of the normal saline treatment group were injected 0.1 ml of normal saline and that of the rhaFGF group were given 4 microg of rhaFGF immediately after reperfusion. The content of D-lactate in the plasma was determined and the changes in intestinal pathology were observed. At the same time, the rates of apoptosis of intestinal epithelial cells were assessed 2, 6, 12 and 24 hours after I/R, and compared to the sham operation group. RESULTS: The plasma content of D-lactate in the saline treatment group at 6 hours after I/R reached (0.34+/-0.09) mg/L and was significantly higher than that in the rhaFGF treatment group((0.23+/-0.07)mg/L, P<0.05). It was shown histologically that the intestinal structures were damaged more seriously in saline treatment group than in rhaFGF group. The apoptosis rates in the saline treatment group and rhaFGF group were elevated significantly, peaking at 12 hours after I/R injury((62.8+/-1.7)% in saline group and (42.5+/-2.6)% in rhaFGF treatment group), then the rate began to fall. There was statistically significant difference between the two groups at 12 hours after I/R injury. CONCLUSION: rhaFGF can reduce content of D-lactate in the plasma and rate of apoptosis of epithelial cells in the intestine after I/R injury, thus it seems to afford a protective effect on the small intestine after I/R injury in rats.

[Effects of reconstructive human acidic fibroblast growth factor and wild type acidic fibroblast growth factor on skin cell proliferation].

OBJECTIVE: To investigate the effects of reconstructive human acidic fibroblast growth factor (aFGF) and wild type aFGF on skin cell proliferation in rat. METHODS: Neonatal rat skin (area of 2 mmx2 mm) was cultured in Dulbecco's modification of Eagle's medium containing reconstructive human aFGF and wild type aFGF, respectively. The concentrations of aFGF were 1 microg/L, 10 microg/L, and 100 microg/L. After being cultured for 4 days, the area of skin was measured. RESULTS: After treatment with two different growth factors in three different concentrations (1 microg/L, 10 microg/L and 100 microg/L) for 4 days, the areas of skin in three reconstructive human aFGF groups were 1.4, 1.5 and 1.3 fold of that of control, respectively and the areas of three wild type aFGF groups were 1.5, 3.2 and 1.6 fold of that of control, respectively, while the area of skin in the control group was (2.96+/-1.12) mm(2). In comparison with those of other groups, the skin area of 10 microg/L wild type aFGF group was significantly increased (P<0.05). CONCLUSION: Reconstructive human aFGF confers less impact on cutaneous cell growth. The capability of wild type aFGF to induce cutaneous cell proliferation is much greater than that of reconstructive human aFGF.

[Effects of acidic fibroblast growth factor on intestinal mucosal damage due to ischemia/reperfusion].

OBJECTIVE: To investigate the effect of exogenous acidic fibroblast growth factor (aFGF) on ischemia/reperfusion (I/R) injury of the intestine in rat, and to explore the mechanism of its protective effect. METHODS: The model of rat intestinal I/R injury was replicated by clamping the superior mesenteric artery (SMA) for 45 minutes followed by reperfusion. Seventy-eight Wistar rats were divided randomly into sham-operation group, intestinal I/R control group and aFGF treated group. In sham-operation group, SMA were separated but not occluded. Animals were sacrificed, samples of blood and tissue from the intestine were obtained 45 minutes after ischemia in sham-operation group and 0.5, 1, 2, 6, 12, and 24 hours after reperfusion in the other groups. Plasma D-lactate level was measured and rat survival rate was noted. Histological changes in intestinal tissue were observed under light microscope. RESULTS: The results showed that rat survival rate in aFGF treatment group was higher than that of intestinal I/R control group (P<0.05), and plasma D-lactate levels were markedly lower in aFGF group than that of intestinal I/R control group (P<0.05). Improvement in pathological changes was observed at 1, 2, 6, 12, and 24 hours after the reperfusion in aFGF treatment group compared with intestinal I/R control group. CONCLUSION: These data suggest that aFGF has a protective effect on intestinal I/R damage, and it may be related to the mitogenic effects and some hormone-like activities of aFGF.

Protective effect of shen-fu on myocardial ischemia-reperfusion injury in rats.

The present study used in vivo rat heart to investigate (1) whether Shen-Fu (SF), a traditional Chinese formulation comprising Radix Ginseng (RG) and Radix Aconitum Carmichaeli (AC), is protective against myocardium damage due to ischemia-reperfusion injury, and (2) whether the cardioprotective effect of SF is related to scavenging of hydroxyl radicals. The model of ischemia-reperfusion injury was established by ligation of left anterior descending coronary artery for 60 minutes followed by reperfusion for 240 minutes in anesthetized rats. The size of infarction and the pathologic changes of myocardium were observed. Lactate dehydrogenase (LDH) and creatine kinase (CK) in serum, the amounts of malondialdehyde (MDA) and superoxide dismutase (SOD) in myocardium were measured at the end of the reperfusion period. Pretreatment groups with SF (10 mg/kg), RG (9 mg/kg) and AC (1 mg/kg) inhibited the rise in MDA and LDH as well as CK, increased SOD activity, reduced the size of infarction, and improved the pathologic changes of myocardium during ischemia-reperfusion compared with the control group. The effect of SF is better than that of RG and AC. These results indicate that SF, RG and AC protect obviously myocardium against damage due to ischemia-reperfusion in rats. The cardioprotective effect of SF injection may be in part related to scavenging of hydroxyl radicals or inhibition of lipid peroxidation. SF is more effective than its separated herbal extracts prepared from RG and AC.

Gonadectomy affects brain derived neurotrophic factor in rats after chronic constriction nerve injury.

AIM: To assess the effect of gonadectomy on brain derived neurotrophic factor (BDNF) expression in neuropathic pain. METHODS: Using chronic constriction injury (CCI) model, we detected BDNF mRNA in dorsal root ganglion and protein content in spinal cord by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay respectively. The time point we chose was post CCI operation d 0, 3, 7, 14, and 21. RESULTS: After CCI surgery, BDNF mRNA in ipsilateral DRGs was upregulated and reached its maximum on post operation d 7. BDNF protein level in ipsilateral spinal cord was also increased and reached its maximum on post operation d 14. The magnitude of this increase in gonadectomy (GDX) rats was significantly smaller than the GDX-sham rats at each time point. CONCLUSION: Gonadectomy reduced the BDNF increment after CCI surgery. Estrogen may affect nociceptive processing by its effect on BDNF.

[Effects of acidi fibroblast growth factor on hepatic and renal functions after intestinal ischemia/reperfusion injury].

OBJECTIVE: To investigate the change in hepatic and renal functions parameters after intestinal ischemia-reperfusion (I/R) injury, and to explore the effects of acidic fibroblast growth factor (aFGF) on hepatic and renal functions after intestinal I/R injury in rats. METHODS: Seventy-eight Wistar rats were divided into four groups, which are sham-operated (C) group, ischemia (45 minutes) plus reperfusion (R), reconstructive human aFGF treatment (rhF), and wild type aFGF treatment (wtF) groups. The animals were sacrificed at 2, 6, 12 and 24 hours, respectively. Hepatic and renal functions were analyzed. RESULTS: In comparison with those in group C, the hepatic and renal functions were damaged in group R, rhF and wtF decreased. Treatment with rhF and wtF markedly abated the hepatic and renal dysfunction. The desquamation of intestine villi and infiltration of inflammation cells in the submucosa were observed in all groups. CONCLUSION: Hepatic and renal functions are damaged after intestinal I/R injury. Treatment with rhF and wtF could protect against multiple organ dysfunction associated with intestinal ischemia-reperfusiun injury.