Objective To investigate the effects of exogenous hydrogen sulfide on brain edema and injury and their mechanisms. Methods Sixty male SD rats were randomly divided into a sham operation group, an ischemia-reperfusion group, a 30 ppm hydrogen sulfide group, and a 60 ppm hydrogen sulfide group (n =15 in each group; 1 ppm =1 mg/L). A model of focal cerebral ischemia for 2 h and reperfusion for 24 h was induced by middle cerebral artery occlusion. The neurological scores were observed after 24 h cerebral ischemia-reperfusion. The cerebral infarction volume, the degree of brain edema, and the changes of blood-brain barrier permeability were measured. Western blotting was used to detect the expressions of occludin and zonula occludens-1 protein (ZO-1) in ischemic penumbra. Results Compared with the ischemia-reperfusion group, the neurological function scores in the 30 ppm and 60 ppm hydrogen sulfide group significantly increased in a dose-dependent manner (al P <0.05) and the neurological deficits and infarct volume reduced (al P < 0.05), and the
brain edema aleviated (al P < 0.05). The content of Evans blue in the ischemic brain tissue in the ischemia-reperfusion group increased significantly compare with the sham operation group (0.74 ±0.14 μg/100 mg vs. 0.19 ±0.06 μg/100 mg; P <0.05). The content of Evans blue in the brain tissue in the 30 ppm hydrogen sulfide group (0.55 ±0.10 μg/100 mg ) and the 60 ppm hydrogen sulfide group (0.35 ±0.08 μg/100 mg ) decreased significantly compared with the ischemia-reperfusion group (al P < 0.05), among them the 60 ppm hydrogen sulfide group was significantly lower than the 30 ppm hydrogen sulfide group (P <0.05). Western blot analysis showed that expression levels of occludin in penumbra (0.621% ±0.101% vs.0.787% ±0.087% vs.0.453% ± 0.127%; P <0.05) and ZO-1 (0.602% ±0.118% vs.0.778% ±0.805% vs.0.426% ±0.146; P <0.05) in the 30 ppm and 60 ppm hydrogen sulfide groups increased significantly compared with the ischemia-reperfusion group, among them, the expression levels of occludin and ZO-1 in the 60 ppm hydrogen sulfide group were significantly higher than those in the 30 ppm hydrogen sulfide group (al P < 0.05). Conclusions Inhalation of exogenous hydrogen sulfide can significantly attenuate brain edema after ischemia-reperfusion in a dose dependent manner, reduce infarct volume, and improve neurological function.Their mechanisms may be associated with inhibiting the downregulated expressions of occludin and ZO-1 and maintaining the integrity of the blood-brain barrier.

Objective: To investigate the effect of RNA interference (RNAi) targeting AKT1 and PI3K P85 on the proliferation and invasion of breast carcinoma MCF-7 cells. Methods: The recombinant adenovirus expression vector, which contained short hairpin RNA (shRNA) targeting open reading frames of AKT1 and PI3K P85 (rAd5-siAKT1-siPI3K), was transfected into human breast carcinoma MCF-7 cells. AKT1 and PI3K P85 mRNA and protein expressions were detected by real-time PCR and Western blotting analysis. The expressions of PCNA, cyclinD1, and P53 were also detected by Western blotting analysis. The proliferation and apoptosis of MCF-7 cells were measured by MTT, flow cytometry and 2-dementinal and 3-dementional matrigel assay. Results: Recombinant adenovirus vector rAd5-siAKT1-siPI3K dramatically down-regulated AKT1 and PI3K P85 mRNA and protein expressions in MCF-7 cells; the downstream factors PCNA and cyclin D1 were also down-regulated, while P53 was up-regulated. Growth of MCF-7 cells was inhibited by over 50% in rAd5-siAKT1-siPI3K group as measured by MTT assay, and cell cycle was arrested in G_1/G_0 phase compared with untransfected and rAd5-siCtrl transfected groups. Cell growth on matrigel matrix showed normal cell shapes, while the cells in rAd5-siAKT1-siPI3K transfected group were detached from the matrix or grew in scattered clustering patterns, forming only small aggregates. Conclusion: shRNA targeting AKT1 and PI3K P85 can significantly down-regulate the expression of AKT1 and PI3K P85 in breast carcinoma MCF-7 cells, and inhibit the growth of MCF-7 cells in vitro.

AIM: To discuss the protective effects and possible mechanisms of 17β-estradiol on human retinal pigment epithelial ( RPE) cells induced by high glucose. ·METHODS: RPE cells were cultured and divided into four groups according to randomized controlled method:blank control group:the cells were treated with 5. 5mmol/L routine glucose medium for processing; high glucose group: cells were treated with 100mmol/L glucose for 12h;17β-estradiol low concentration group: after treated with 10 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h; 17β-estradiol high concentration group: after treated with 100 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h. Cell viability were tested by MTT colorimetric detection. Cells apoptosis were detected by Hochest33258 staining. Intracellular reactive oxygen species( ROS) level were detected by H2 DCFDA staining. Expression of CAT, SOD and MDA were tested by colorimetric detection. · RESULTS: RPE cell activity decreased with the concentration of glucose increased; 17β-estradiol inhibited high glucose-induced cell viability decrease in RPE cells, decreased the apoptosis rate of RPE cells and intracellular ROS generation; besides, 17β-estradiol significantly increased the expression of CAT, SOD and decreased the expression of MDA in RPE cells. ·CONCLUSION: The 17β-estradiol effectively inhibited high glucose -induced RPE cells damage, which provide reliable experimental basis for the treatment of injuries in RPE cells.

An anti-alpha-synuclein (alpha-SYN) monoclonal antibody produced in our laboratory was used to investigate the effect of repeated acute hypoxic treatments on the expression of alpha-SYN in the mouse cerebral cortex. Western blot analysis showed that the expression levels of alpha-SYN in the cortex changed accordingly upon hypoxic exposure times, as that the alpha-synuclein level significantly increased after the first hypoxic exposure and then dropped down to the background level after the fourth hypoxic exposure. Immunohistochemical staining revealed that the alpha-SYN-immunopositive substance was localized not only in the nerve endings, but also within the nuclei of some neurons. The cell density of the neurons with alpha-SYN immunopositive nuclei was increased significantly after the first hypoxic exposure but returned back to control levels after the fourth hypoxic exposure. Our results indicate that both of the alpha-SYN expression level in the brain and the number of the neurons with alpha-SYN positive nuclei are affected by the repeated acute hypoxic treatments and that this modification is hypoxic time-dependent. The mechanism and the physiological significance underlying these changes need to be further investigated.
Animals ; Brain ; blood supply ; Brain Ischemia ; metabolism ; Cerebral Cortex ; metabolism ; Ischemic Preconditioning ; Mice ; Mice, Inbred BALB C ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Neurons ; metabolism ; Phosphoproteins ; biosynthesis ; genetics ; Random Allocation ; Synucleins ; alpha-Synuclein

OBJECTIVEThe aim of this study was to evaluate the value of sixteen-detector row computed tomography angiography (CTA) for the assessment of coronary artery bypass graft (CABG).

METHODSSixty-two consecutive patients undergoing coronary artery bypass grafting were recruited. Among them, 6 patients were excluded from the study due to unfavorable control of heart rate. A total of 56 patients with 152 coronary artery bypass grafts (internal mammary artery, n = 48; saphenous venous grafts, n = 104) were examined by computed tomography angiography (CTA) with sixteen-detector row CT and by conventional invasive coronary angiography (CAG). All CT procedures were performed with retrospective electrocardiogram gating method. The patients' mean heart rate was 58 +/- 6 beats/minute. 120 ml of Visipaque 320 were continuously injected with the rate of 4.0 ml/sec during the procedure. The patency and the stenosis of coronary artery bypass grafts were evaluated by two experienced readers.

RESULTSAll the coronary artery bypass grafts were visualized by CTA, and all the proximal bypass anastomoses and 71% of the distal bypass anastomoses were also visualized by CTA. Furthermore, 29 occlusions and 13 significant stenoses of coronary artery bypass grafts were detected by CTA. The comparison of the results between CTA and CAG showed that among all the 42 occluded and stenosed coronary artery bypass grafts detected by CTA, 34 were confirmed by CAG; among all the 110 normal coronary artery bypass grafts detected by CTA, 108 were confirmed by CAG. There were 8 false positive and 2 false negative findings, resulting in a sensitivity of 94%, a specificity of 95%, a positive predictive value of 86%, and a negative predictive value of 99%.

CONCLUSIONSixteen-detector row CTA technology may provide a reliable visualization and higher diagnostic accuracy of coronary artery bypass grafts lesions. This technique can be used as a noninvasive procedure for the diagnosis of suspected coronary artery bypass grafts dysfunction.

Aged ; Coronary Angiography ; methods ; Coronary Artery Bypass ; Coronary Artery Disease ; diagnostic imaging ; surgery ; Coronary Restenosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed

OBJECTIVETo summarize the characteristics and treatment of burn casualties of yellow phosphorus explosion, so as to share the experiences in emergency treatment.

METHODSBy analyzing the data related to this accident, the characteristics of the injury and experiences of treatment for mass burn casualties from yellow phosphorous explosion were summarized.

RESULTSEighty-one patients, 72 males and 9 females, were injured in a yellow phosphorus explosion. The mean age of the patients was 24 +/- 13 years old (5-42 y). The mean total burn surface area was (9 +/- 11)% [(0.4% - 70.0%))] TBSA, and the mean burn surface area of III degrees/IV degrees was (7 +/- 10)% [(0.4% - 60.0%)] TBSA. Most of the patients showed the symptoms and signs of phosphorus poisoning. Among all the patients, 27 cases (33.3%) showed hepatic dysfunction, 15 cases (18.5%) had renal dysfunction, 42cases (51.9%) showed electrolytes disorders. Among the 8 patients with burn surface area over 10% TBSA and less than 20% TBSA, high levels of cardiac enzymes were found in 6 cases, anaemia in 7 cases (3 with progressive anaemia), asphyxia occurred in 1 case 48 hours after burn, and in 1 case complicated with stress ulcer. Escharectomy and skin grafting were performed within four days after burn in 72 patients. All the patients survived, some of them showed impaired hand function and hypertrophic scar, and partial finger amputation was done in 3 patients.

CONCLUSIONYellow phosphorus explosion produces deep burn injuries in surrounding people especially in exposed parts such as head, hand and so on. Adequate organization of medical resources for emergency treatment, early debridement, and accelerating excretion of phosphorus are the key points for the successful rescue of mass casualties.

Accidents, Occupational ; Adolescent ; Adult ; Burns, Chemical ; etiology ; therapy ; Child ; Child, Preschool ; Combined Modality Therapy ; Female ; First Aid ; Humans ; Male ; Phosphorus ; adverse effects ; Young Adult

BACKGROUNDIntravenous urography (IVU) combined with add-on CT (IVU-CT) can help to provide more diagnostic information for determining the localization and nature of ureteral abnormalities with less irradiation dose. This study aimed to determine the value of IVU-CT for diagnosis of ureteral diseases, where IVU is insufficient to determine the diagnosis.

METHODSTwo hundred and eighty patients underwent IVU for suspected ureteral disorders, which identified a definite diagnosis in 184 cases and was insufficient for definite diagnosis in 96 cases designated as indeterminate diagnosis. Subsequently 90 patients (six patients declined CT) with indeterminate diagnosis consented to undergo immediate or delayed helical CT scan. The CT data were transferred to the workstation for post-processing, and the cost and mean effective dose for each imaging method were calculated and compared indirectly.

RESULTSOf the 90 indeterminate diagnosis cases, diagnosis was determined in 86 cases by IVU-CT with a diagnostic accordance rate of 95.6%, while 184/280 (65.7%) had diagnosis determined by IVU alone. There was a significant difference between IVU and IVU-CT in the determination of the diagnosis of ureteral diseases (c(2) = 36.4, P < 0.05). The cost of IVU equals to 1/8 - 1/9 of that for CT urography (CTU), and the cost of IVU-CT is as much as 1/3 of CTU. CTU results in the highest mean effective dose, approximately nine times that for IVU and three times that for IVU-CT.

CONCLUSIONIVU-CT provides valuable information for the localization and diagnosis of ureteral abnormalities and may be considered as an efficient, cost-effective and low-dose diagnostic technique in this setting.

Adolescent ; Adult ; Aged ; Child ; Cost-Benefit Analysis ; Female ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; economics ; methods ; Ureter ; diagnostic imaging ; Ureteral Diseases ; diagnosis ; Urography ; economics ; methods ; Young Adult

OBJECTIVETo clone the recombinant human islet neogenesis-associated protein (rhINGAP) gene for its secretory expression in Pichia pastoris.

METHODSINGAP gene was amplified with PCR and inserted between Xho I and EcoR I downstream sites of the alpha factor of the recombinant plasmid alpha/pUC18. The fusion gene of alpha factor and INGAP was subsequently inserted between BamH I and EcoR I sites of the plasmid pPIC9K of P. pastoris. After confirmation with restriction enzyme digestion and sequencing, the positive recombinant plasmid that integrated INGAP gene was linearized with Sal I digestion and transformed into the yeast host strain GS115 through electroporation. The yeast transformants that harbored the INGAP gene with high copies were selected with the auxotroph medium and G418, followed then by PCR verification of the positive transformants, from which the expression of recombinant human INGAP was induced with methanol as the only carbone source. The antigenic activity of the desired protein was then detected using Western blotting and enzyme-linked immunosorbent assay (ELISA).

RESULTS AND CONCLUSIONThe recombinant expression plasmid INGAP/pPIC9K was successfully constructed, and 3 positive transformants were obtained. The expressed protein showed good antigenic activity as confirmed by Western blotting and ELISA.

Antigens, Neoplasm ; genetics ; metabolism ; Biomarkers, Tumor ; genetics ; metabolism ; Blotting, Western ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Humans ; Islets of Langerhans ; metabolism ; Lectins, C-Type ; genetics ; metabolism ; Pancreatitis-Associated Proteins ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; metabolism

BACKGROUNDLaparoendoscopic single-site surgery radical prostatectomy (LESS-RP) is a challenging urological procedure and needs to be further evaluated. This study was undertaken to illustrate the safety and initial results of pure LESS-RP with conventional available instruments.

METHODSA prospective clinical database was established in September 2010 to assess the outcome following the introduction of LESS-RP at our institution. By June 2012, six procedures had been performed. The mean patient age was 74.7 (74.0 - 76.0) years. The mean body mass index (BMI) was 23.8 (19.5 - 32.2) kg/m(2). The LESS-RP was preformed through an extra-peritoneal approach using single port access with QudaPort, 0° lens 5 mm flexible tip video-laparoscope and available conventional laparoscopic instruments. Parameters assessed were operative time, estimated blood loss, intra-operative complications, drainage time, postoperative pain score (visual analogue pain scale (VAPS), 0 - 10), pathological results, and postoperative prostate specific antigen (PSA) levels.

RESULTSLESS-RP was completed for all six cases without additional trocar placement or conversion to standard laparoscopic or open radical prostatectomy. The mean operative times were 252.5 (190.0 - 305.0) minutes, estimated blood loss was 300 (100 - 500 ml). There was no documentable intraoperative complication. The mean wound-drainage time was 5.2 (2.0 - 7.0) days, and the first postoperative day VAPS was 0.8 (0 - 3.0). Final pathological staging was pT2aN0M0 in four cases and pT2cN0M0 in two cases. Surgical margins were negative for all cases. The one-month post-operative PSA was less than 0.02 ng/ml in each case. All patients were continent without pad usage at 1 year postoperatively.

CONCLUSIONThe technique of pure LESS-RP is feasible and early outcomes are acceptable.

Aged ; Humans ; Laparoscopy ; methods ; Male ; Prospective Studies ; Prostate-Specific Antigen ; blood ; Prostatectomy ; methods ; Prostatic Neoplasms ; blood ; surgery ; Treatment Outcome

OBJECTIVETo compare the differences of the clinical effects, side effects and treatment-related cost between two kinds of negative-pressure wound therapy (NPWT).

METHODSForty-four inpatients with acute, subacute, and chronic wounds were divided into simplified NPWT group (A group) and conventional NPWT group (B group) according to the random number table. Wounds of patients in A group were treated with gauze + continuous suction with hospital central negative pressure (-10.64 kPa) for 24 hs; wounds of patients in B group were treated with sponge + interrupted suction with a purpose-designed suction appliance (-16.63 kPa) for 24 hs. Gross wound condition, treatment time, survival rates of skin graft and flap, changes of bacterial species on wound, treatment cost, and ratio of side effects between two groups were compared.

RESULTSThere was no significant difference between A and B groups in respect of gross wound condition, treatment time [A group (29 +/- 12) d, B group (26 +/- 13) d, P > 0.05], changes of bacterial species, survival rates of skin graft [A group (98 +/- 4)%, B group (98 +/- 4)%, P > 0.05] and flap (A group 98%, B group 100%, P > 0.05). Treatment cost of A group yen(374 +/- 134) was obviously lower than that of B group yen(9825 +/- 4956) (P < 0. 01), while more side effects were observed in A group (33.3%) than that in B group (5.0%) (P < 0.05).

CONCLUSIONSBoth simplified NPWT and NPWT with purpose-designed appliance can effectively improve wound healing. The simplified method may cause many side effects and has a potential risk of inciting nosocomial infection, but it can be conveniently employed with a low cost. In contrast, the cost of using purpose-designed appliance should be cut down to meet the aim of generalization.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; methods ; Wound Healing ; Young Adult