To study the correlation between the spatial cognitive impairment and different subtypes of estrogen receptor α (ERα) of hippocampus in diabetic mice, we used alloxan (intraperitoneal injection) to induce type 1 diabetes in male Kunming mice and compared the spatial cognitive ability of the model mice with that of control mice through Morris water maze test. Meanwhile, using Western blot, we detected the protein expressions of ER-α36, ER-α66, caveolin-1, PKCα, cAMP-response element binding protein 2 (CREB2), and synaptophysin (Syn) in the hippocampus of the mice. The results showed that on the 3rd and 5th days of training, the ability of spatial learning and memory in the diabetic mice was significantly inferior to that of the control mice (P < 0.05). In the diabetic mice, the protein expressions of caveolin-1 and PKCα were decreased (P < 0.05), but ER-α66 expression was unaffected, while ER-α36 and CREB2 expressions were significantly increased (P < 0.05) compared with those of the control mice. The results suggest that abnormal expression of ER-α36 and related signal molecules may be important factors for diabetes-induced spatial cognitive impairment.
Animals ; Caveolin 1 ; metabolism ; Cognitive Dysfunction ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Diabetes Mellitus, Experimental ; physiopathology ; Estrogen Receptor alpha ; metabolism ; Hippocampus ; metabolism ; physiopathology ; Male ; Maze Learning ; Memory ; Mice ; Protein Kinase C-alpha ; metabolism ; Synaptophysin ; metabolism

OBJECTIVETo investigate the mechanism of ethanol-induced calcium overload in hepatocytes and the related role of store-operated calcium channels (SOCs).

METHODSHepG2 cells were treated an ethanol concentration gradient with or without intervention treatment with the extracellular calcium chelator EGTA or the SOCs inhibitor 2-aminoethoxydiphenyl borate (2-APB). Effects on cell viability were assessed by the CCK8 assay. Effects on leakage of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined by automatic biochemical analyzer measurements of the culture supernatants. Effects on cytoplasmic free Ca2+ concentration ([Ca2+]i) were accessed by detecting fluorescence intensity of the calcium indicator Fluo-3/AM with a flow cytometer. Effects on mRNA and protein expression levels of SOCs, stromal interacting factor 1 (STIM1), and calcium release-activated calcium channel protein 1 (Orai1) were evaluated by qPCR and western blotting.

RESULTSThe ethanol treatment produced dose-dependent reduction in cell viability (r = -0.985, P less than 0.01) and increases in leakage of ALT (F = 15.286, P less than 0.01) and AST (F = 39.674, P less than 0.01). Compared to untreated controls, the ethanol treatments of 25, 50, 100, 200 and 400 mM induced significant increases in [Ca2+]i level (1.25+/-0.36, 1.31+/-0.15, 1.41+/-0.18, 2.29+/-0.25, 2.58+/-0.19; F = 15.286, P less than 0.01). Both intervention treatments, EGTA and 2-APB, significantly reduced the 200 mM ethanol treatment-induced [Ca2+]i increase (2.32+/-0.08 reduced to 1.79+/-0.15 (t = 7.201, P less than 0.01) and 1.86+/-0.09 (t = 8.183, P less than 0.01) respectively). EGTA and 2-APB also increased the ethanol-treated cells' viability and reduced the ALT and AST leakage. The 200 mM ethanol treatment stimulated both gene and protein expression of STIM1 and Orai1, and the up-regulation effect lasted at least 72 h after treatment.

CONCLUSIONEthanol-induced dysregulation of SOCs may be an important molecular mechanism of ethanol-induced [Ca2+]i rise in hepatocytes and the related liver cell injury.

Calcium ; metabolism ; Calcium Channels ; metabolism ; Ethanol ; adverse effects ; Hep G2 Cells ; Hepatocytes ; drug effects ; metabolism ; Humans

OBJECTIVETo investigate the role of store-operated calcium channels (SOCs) in primary hepatocytes under conditions of calcium overload and ethanol-induced injury.

METHODSThe in vitro model of chronic ethanol-induced hepatocyte injury was established using primary hepatocytes isolated from Sprague-Dawley rats. Ethanol-induced changes (24, 48 and 72 h; 50, 100, 200, 400 and 800 mmol/L) in expression of the SOCs proteins stromal interaction molecule 1 (STIM1) and calcium release-activated calcium channel protein 1 (Oria1) were detected by qualitative PCR analysis (mRNA) and western blotting (protein). The possible role of these two SOCs proteins in the ethanol-induced extracellular calcium influx and related liver cell injury was determined by treating the cell system with various channel blockers (EGTA, La3+, and 2-APB). Cell viability was determined by MTT assay and cytosolic free calcium ion concentration was determined by flow cytometry.

RESULTSAfter 24 h of exposure to 0 (untreated) to 800 mM/L ethanol, the cell viability was reduced in a concentration-dependent manner. The 400 mmol/L concentration of ethanol decreased cell viability by 57.34% +/- 2.34%. and was chosen for use in subsequent experiments. Compared with the untreated control cells, the ethanol-treated cells showed significantly up-regulated mRNA and protein expression of both STIM1 and Orai1 at all times examined, suggesting that the ethanol-stimulated expression of STIM1 and Orai1 could persist for at least 72 h. The ethanol treatment induced increase in cytoplasmic calcium levels was significantly (and similarly) reduced by co-treatment with any of the three channel blockers.

CONCLUSIONChronic ethanol exposure can increase the expression of STIM1 and Orai1 in primary liver cells, suggesting that ethanol may increase extracellular calcium influx by up-regulating expression of these SOCs protein molecules, ultimately aggravating liver cell damage.

Animals ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Calcium Channels ; metabolism ; Calcium-Binding Proteins ; metabolism ; Cell Survival ; Cells, Cultured ; Ethanol ; adverse effects ; Hepatocytes ; drug effects ; metabolism ; Male ; Membrane Glycoproteins ; metabolism ; Primary Cell Culture ; Rats ; Rats, Sprague-Dawley ; Stromal Interaction Molecule 1

OBJECTIVETo analyze the clinical features and survival time in primary myelodysplastic syndromes (MDS) patients accompanied with immunological abnormalities.

METHODSThe clinical information, laboratory findings and survival time in 194 untreated primary MDS patients with complete immunological laboratory tests or a past history of autoimmune disease were analyzed retrospectively.

RESULTSThere were 37/194 cases (19.07%) with autoimmune abnormalities, including 16/194 (8.25%) with autoimmune disease and 21/194 asymptomatic cases (10.82%) with serologic immunological abnormalities only. There was significant differences in the distribution of age < 60 years old, female, CD4(+)T-cell/CD8(+)T-cell ration < 1 and trisomy 8 (P < 0.05) between the cases with autoimmune disease and without autoimmune abnormalities. The former had a higher 2-year OS, but there was no significance (P = 0.065). There was no significant differences in the distribution of age, MDS-subtype, IPSS risk groups, haemoglobin, absolute neutrophil count, platelets count, the severity of anemia and neutropenia, high level of serologic TNF, chromosomal abnormalities, cytogenetic risk groups and bone marrow cellularity (P > 0.05).

CONCLUSIONMDS patients with autoimmune disease are mainly female and younger than 60 years old, with high proportion of trisomy 8 and better prognosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Autoimmune Diseases ; complications ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; complications ; mortality ; Retrospective Studies ; Survival Analysis ; Young Adult

OBJECTIVETo evaluate the effect of surgical bypass and to explore the role of transcranial doppler (TCD) during the bypass operation.

METHODSFrom March 2003 to February 2008, sixteen patients (4 male and 12 female) with mean age of 32 years old and mean disease course of 7.5 years were treated by surgical procedures. The main clinical presentations were dizziness, headache, vertigo, and visional dysfunction. Variated degree of artery stenoses in the 4 arteries (bilateral carotid and vertebral arteries) were revealed by color doppler ultrasonography and DSA. Eight patients underwent aorto-bi-subclavian arteries prosthetic graft bypass. Three patients underwent aorto-bi-subclavian artery prosthetic graft bypass and graft-unilateral carotid artery bypass with autologous great saphenous vein. Three patients underwent aorta-unilateral subclavian artery-unilateral carotid artery prosthetic graft bypass. Two patients underwent aorta carotid artery bypass with autologous great saphenous vein, of which one patient underwent aortocoronary artery bypass simultaneously. Cerebral blood supply was monitored in 14 patients with transcranial doppler. Unilateral subclavian carotid and femoral carotid artery shunt was used respectively to avoid cerebral ischemia during operation in 2 patients.

RESULTSSymptoms and signs of cerebral ischemia improved in all patients with effective rate of 100% apart from deflected tongue-protrusion in 3 patients which recovered in 2 weeks after operation. All patients survived and no symptoms recurred at the end of a 2.2 year's follow-up. Unfortunately, two patients developed aneurysm at the anastomosis within 4 years after operation.

CONCLUSIONSArterial reconstruction is an effective method for Takayasu's disease with severe cerebral ischemia. TCD monitoring plays an important role during the bypass operation. It can help to determine the revision of blood pressure and prevent postoperative brain reperfusion injury.

Adult ; Brain ; blood supply ; Brain Ischemia ; etiology ; surgery ; Cerebral Revascularization ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; Regional Blood Flow ; Takayasu Arteritis ; complications ; surgery ; Treatment Outcome ; Ultrasonography, Doppler, Transcranial ; Young Adult

OBJECTIVETo investigate the management of complicated, severe or recurrent Budd-Chiari syndrome.

METHODSFrom February 2004 to August 2007, 28 patients with complicated, severe or recurrent Budd-Chiari syndrome were treated. In this series, 16 patients relapsed after treated with percutaneous transluminal angioplasty or stent deployment, 2 cases relapsed after surgery; and the other 10 were under severe conditions and hard to treat, including malignancy of the inferior vena cava and right atrium. Meso-cavo-atrial shunt was carried out in 10 cases, meso-cavo-jugular shunt in 6 (capitis medusa was used in one case), cavoatrial shunt in 2 and cavo-jugular shunt in 1, mesocaval shunt in 2, and radical or extended radical correction in 7.

RESULTSOne patient (3.6%) died in 24 hours after operation. Graft infection occurred in 1 case. Excellent, good, fair, poor and death rate were 22.2%, 55.5%, 14.8%, 3.7% and 3.7%, respectively, the overall effective rate was 92.5%.

CONCLUSIONTo select personalized treatment according to the disease status brings hopes to difficult, severe, recurrent Budd-Chiari syndrome.

Adolescent ; Adult ; Blood Vessel Prosthesis Implantation ; Budd-Chiari Syndrome ; surgery ; Child ; Child, Preschool ; Critical Illness ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Portacaval Shunt, Surgical ; Recurrence ; Retrospective Studies ; Treatment Outcome

BACKGROUNDMany treatment options for lower limb ischemia are difficult to apply for the patients with poor arterial outflow or with poor general conditions. The effect of medical treatment alone is far from ideal, especially in patients with diabetic foot. A high level amputation is inevitable in these patients. This study aimed to explore the effect of transplantation of autologous bone marrow mononuclear cells on the treatment of lower limb ischemia and to compare the effect of intra-arterial transplantation with that of intra-muscular transplantation.

METHODSIn this clinical trial, 32 patients with lower limb ischemia were divided into two groups. Group 1 (16 patients with 18 affected limbs) received transplantation of autologous bone marrow mononuclear cells by intra-muscular injection into the affected limbs; and group 2 (16 patients with 17 affected limbs) received transplantation of autologous bone marrow mononuclear cells by intra-arterial injection into the affected limbs. Rest pain, coldness, ankle/brachial index (ABI), claudication, transcutaneous oxygen pressure (tcPO(2)) and angiography (15 limbs of 14 patients) were evaluated before and after the mononuclear cell transplantation to determine the effect of the treatment.

RESULTSTwo patients died from heart failure. The improvement of rest pain was seen in 76.5% (13/17) of group 1 and 93.3% (14/15) of group 2. The improvement of coldness was 100% in both groups. The increase of ABI was 44.4% (8/18) in group 1 and 41.2% (7/17) in group 2. The value of tcPO(2) increased to 20 mmHg or more in 20 limbs. Nine of 15 limbs which underwent angiography showed rich collaterals. Limb salvage rate was 83.3% (15/18) in group 1 and 94.1% (16/17) in group 2. There was no statistically significant difference in the effectiveness of the treatment between the two groups.

CONCLUSIONSTransplantation of autologous bone marrow mononuclear cells is a simple, safe and effective method for the treatment of lower limb ischemia, and the two approaches for the implantation, intra-muscular injection and intra-arterial injection, show similar results.

Aged ; Aged, 80 and over ; Blood Gas Monitoring, Transcutaneous ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Female ; Humans ; Ischemia ; therapy ; Leg ; blood supply ; Leukocytes, Mononuclear ; transplantation ; Male ; Middle Aged ; Transplantation, Autologous

OBJECTIVETo summarize therapeutic efficacy of vascular reconstruction in treating infrapopliteal arterial occlusion.

METHODSRetrospective analysis of vascular reconstruction of lower extremity was made in 56 cases suffering from popliteal arteries or 3 branches of popliteal arteries (anterior tibial, posterior tibial, peroneal artery) between July 2001 and August 2005 in our hospital. According to the level of lower extremity arterial occlusion, a composite grafts which consisting of a combined proximal PTFE prosthesis grafts with autogenous vein grafts were used to establish the sequential vascular reconstruction for multilevel and multistage arterial occlusive disease.

RESULTSThe mean follow-up period after operation was 17 months. The primary graft patency rates in this series was 67.3%, the secondary graft patency rate was 78.8%.

CONCLUSIONSThe treatment of composite sequential bypass is a practical means for multilevel arterial occlusive disease suffering from femoral-infrapopliteal arteries, which effectively resolves the insufficiency supply of autogenous vessel grafts and the problem of bad patency rate for vascular reconstruction with prosthetic grafts alone in arterial occlusion suffering from infrapopliteal arteries.

Aged ; Aged, 80 and over ; Arterial Occlusive Diseases ; surgery ; Blood Vessel Prosthesis Implantation ; Female ; Follow-Up Studies ; Humans ; Leg ; blood supply ; Male ; Middle Aged ; Popliteal Artery ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Saphenous Vein ; transplantation ; Treatment Outcome ; Vascular Surgical Procedures ; methods

Adult ; Aged ; Alcohol Drinking ; China ; ethnology ; Fatty Liver, Alcoholic ; etiology ; Female ; Humans ; Male ; Middle Aged ; Surveys and Questionnaires

OBJECTIVETo study the relationship between the expression of Bax, Bcl-2 proteins, and apoptosis in radiation compound wound healing of rats.

METHODSApoptosis, Bax and Bcl-2 proteins were estimated by in situ terminal labeling (TUNEL) and immunohistochemical methods.

RESULTS(1) Changes of the apoptosis in wound healing showed three typical characteristics: early occurrence, high frequency and delayed disappearance after radiation to rats when compared with those of simple wound group, which might be an important reason for radiation-induced delayed wound healing. (2) The expression of Bax protein increased evidently with the increment of apoptosis and showed a good corresponding relationship with the apoptotic frequency in the process of wound healing. While the expression of Bcl-2 protein decreased obviously as the apoptosis reached a maximum and showed increasing tendency up to normal level when the apoptosis decreased distinctively.

CONCLUSIONSBax and Bcl-2 proteins play an important role in the apoptotic regulation of radiation compound wound healing in rats.

Animals ; Apoptosis ; radiation effects ; Female ; Gamma Rays ; Immunohistochemistry ; Proto-Oncogene Proteins ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Skin ; pathology ; radiation effects ; Wound Healing ; genetics ; radiation effects ; bcl-2-Associated X Protein