High resolution two-dimensional polyacrylamide gel electrophoresis, followed by computer-assisted analysis, was applied to investigate the fat body of silkworm, Bombyx mori. 722 spots were obtained by silver stain from 18 cm, pH 3-10 gel. Most of them were distributed in the area from 15 kD to 90 kD with pH 4 - 8. The matrix-assisted laser desorption ionization time of flight mass spectrometry were applied for identifying the major spots of the 2D map. A total of 41 spots were excised to identify by a combination of MALDI-TOF MS after digested with trypsin. The result showed among the 34 proteins identified, plenty of them were involved in metabolism and immunity. Additionally, Heat shock proteins, 30K proteins and Actin were also detected in the fat body of silkworm. The result will provide a useful tool for understanding the role of fat body in silkworm.
Adipose Tissue ; chemistry ; metabolism ; Animals ; Bombyx ; chemistry ; genetics ; Electrophoresis, Gel, Two-Dimensional ; Proteome ; analysis ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Bletilla striata has been used as traditional Chinese medicine for several centuries. In recent years, the quality and quantity of wild B. striata plants have declined sharply due to habitat deterioration and human over-exploitation. Therefore, it is of great urgency to evaluate and protect B. striata wild plant resource. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to assess the level and pattern of genetic diversity in twelve populations of B. striata. The results showed a high level of genetic diversity (PPB = 90.48%, H = 0.349 4, I = 0.509 6) and moderate genetic differentiation among populations (G(st) = 0.260 9). Based on the unweighted pair-group method with arithmetic average (UPGMA), twelve populations gathered in three clusters. The cluster 1 included four populations. There are Nanjing, Zhenjiang, Xuancheng and Hangzhou. The seven populations which come from Hubei Province, Hunan Province, Jiangxi Province and Guizhou Province belonged to the cluster 2. The cluster 3 only contained Wenshan population. Moreover, Mantel test revealed significant positive correlation between genetic distances and geographic distances (r = 0.632 9; P < 0.000 1). According to the results, we proposed a series of conservation consideration for B. striata.
China ; Genetic Markers ; Genetic Variation ; Genetics, Population ; Orchidaceae ; genetics ; Phylogeny ; Plants, Medicinal ; genetics

We separated proteins of the middle silk gland through high resolution two-dimensional polyacrylamide gel electrophesis, and identified the high expressional proteins using MALDI-TOF-MS and analyzed the PMF in protein database by GPMAW( General Protein/Mass Analysis for Windows) . The protein database was forecasted through silkworm genome database. More than 500 spots were obtained from each gel by silver stain and more than 100 protein spots were obtained from gel by Coomassie brilliant blue stain. Most of them were distributed in the area from 15kD to 90kD with pH 3.5-7. Among the 25 Coomassie brilliant blue stained spots identified by MALDI-TOF-MS, more than 60% have relatively strong signal. Comparing with the result of using Mascot, the method using PMF database of silkworm not only can identify some known proteins, but also can identify some unknown proteins that have been forecasted in silkworm genome database. Accordingly, we founded a complete set of method that fit for researching proteome of silkworm.
Amino Acid Sequence ; Animals ; Bombyx ; metabolism ; Databases, Protein ; Electrophoresis, Gel, Two-Dimensional ; Hydrogen-Ion Concentration ; Insect Proteins ; analysis ; chemistry ; Molecular Sequence Data ; Molecular Weight ; Proteome ; analysis ; chemistry ; Proteomics ; methods ; Silk ; metabolism ; Software ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Adult ; Aged ; Breast Neoplasms ; complications ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Leptin ; metabolism ; Lymph Nodes ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Receptors, Leptin ; metabolism

AIMTo learn the condition of the memory and the visual space of the patient who has suffered from the infarction in basal ganglia region, and to analyze its impact on and the characteristics of the patients' cognition.

METHODSBy testing respectively on 21 subjects who were initially infarcted in basal ganglia with single focus on one side, and 21 healthy volunteers with corresponding age, gender and educational background with Rey-Osterrieth Complex Figure Scale (Rey), Clinical Memory Scale (CMS), Hospital Anxiety-Depression Scale (HAD), National Institute of Health Stroke Scale (NIHSS), Oxford Handicap Scale (OHS), Barthel Index (BI), and Aural Event Related Evoked Potential (AERP) as well.

RESULTSCompared with the control group, the patient group got higher scores of anxiety and depression in HAD), which showed absolutely statistical significance; they demonstrated longer reaction time in AERP, which also showed statistical significance; and in CMS. they displayed inferior performance in the free picture recall and the memory quotient (MQ), which again had statistical significance. However, compared with the control group, the patient group got scores with no significance in copy, immediate recall and delayed recall in Rey.

CONCLUSIONThe infarction in basal ganglia region with single focus on one side may impact on the patient's executive function, memory function and emotion. But no impact is showed on patient's visual space function.

Adult ; Aged ; Basal Ganglia ; pathology ; Brain Infarction ; pathology ; physiopathology ; psychology ; Case-Control Studies ; Evoked Potentials, Auditory ; Female ; Humans ; Male ; Middle Aged ; Neuropsychological Tests

Objective To evaluate the effect of myocardial viability on left ventricular function after elective revascularization in patients with myocardial infarction by 99Tcm-MIBI and 18F-FDG dual-isotope simultaneous acquisition (DISA) myocardial perfusion-metabolic imaging. Methods Ninety-one patients clinically confirmed of myocardial infarction underwent DISA imaging. Based on the results of echocardiography, the patients were divided into heart failure group (group A) and normal cardiac function group (group B). After PCI, left ventricular function was measured by echocardiography in 1, 3 and 6 months. The t-test and χ2-test were used to compare the difference between the two groups using SPSS 13.0. Results The average number of diseased segments by myocardial perfusion imaging was 9.8±3.5 and 5.4±2.6 in groups A and B, respectively (t=6.87, P<0.01). The average number of diseased segments by myocardial metabolic imaging was 7.5±3.4 and 4.6±2.8 in groups A and B, respectively (t=4.46, P<0.01). There were 173 segments with viable myocardium (173/458: 37.8%) in group A and 188 segments with viable myocardium (188/307: 61.2%) in group B (χ2=40.61, P<0.001). The summed perfusion score (SPS), summed metabolism score (SMS) and summed difference score (SDS=SMS-SPS) were 28.43±11.86 vs 21.36±9.54, 20.17±8.52 vs 15.19±5.74 and 0.39±3.17 vs -12.72±4.55, respectively in groups A and B (t=3.15, P<0.01; t=3.32, P<0.01; t=15.59, P<0.01). The mean change of LVEF (ΔLVEF) and the mean change of left ventricular end-diastole dimension (ΔLVEDd) of the patients with more than 4 viable myocardial segments in group A were significantly more than those in group B( (12.81±2.62)% vs (5.90±1.91)%, t=16.33, P<0.001; (-13.13±4.20) mm vs (-7.75±2.31) mm, t=6.86, P<0.001). However, the ΔLVEF and ΔLVEDd of the patients with less than 4 viable myocardial segments in group A were significantly less than those in group B (t=3.25, P<0.01; t=4.92, P<0.001). Conclusion The amount of viable myocardium in infarct myocardium is an important factor for left ventricular function recovery after elective revascularization.

Purpose To observe the effect of Necrostatin-1 (Nec-1),a necroptosis inhibitor,on the inflammation in unilateral ureter obstruction mice.Methods Male C57BL/6J mice were randomly divided into sham operation group,unilateral ureter obstruction operation group and UUO + Nec-1 treatment group,and the mice were sacrificed at 7th day after operated.Scr and BUN were measured.The pathological changes in the kidney were observed by HE staining.Immunohistochemistry and Western blot were used to detect the protein expression of necroptosis-related indicators RIP1,RIP3,MLKL and inflammatory cytokines TNF-α,IL-1β and MCP-1.Results Compared with sham operation group,the expression of RIP1,RIP3 and MLKL protein increased in the renal tissue of UUO mice,accompanied with increased expression of TNF-α,IL-1β and MCP-1.Nec-1 treatment significantly decreased above-mentioned protein expression in UUO mice,and also reduced renal interstitial inflammation and renal tubal injury according to HE staining.Scr and BUN levels suggested improved renal function.Conclusion Nec-1 could relieve the inflammatory reaction in renal tissue of the UUO mice by inhibiting necroptosis,which may be a new target for the treatment of secondary inflammation.

Objective To study the levels of noradrenaline, epinephrine, angiotensin Ⅱ (Ang Ⅱ) and rennin in acute phase of intracerebral hemorrhage (ICH) and their association with the blood pressure regulatory mechanisms in spontaneously hypertensive rats. Methods ICH was induced in spontaneously hypertensive rats by injection of collagenase into the caudate nucleus. Before and at 1, 3,7 and 14 days after ICH, the plasma and hypothalamus noradrenaline and epinephrine levels were determined with high-performance liquid chromatography, and the Ang Ⅱ levels and renin activity were examined by radioimmtmoassay. Results The plasma noradrenalin and epinephrine levels and hypothalamus noradrenalin level increased significantly after ICH, reaching the peak levels at 24 h (4.31± 0.97 μg/L, 8.55±2.41 μg/L, and 919±128 ng/g; P=0.011, 0.015, and 0.005, respectively) and recovered the preoperative levels till 7-14 days later. The peak levels of plasma rennin and Ang Ⅱ and hypothalamus 035, 0.036, and 0.024, respectively), and were lowered to the preoperative levels 7-14 days after ICH. One day after ICH, the blood pressure of the rats showed a significant increase (P=0.000) followed by obvious reduction on day 3 and recovered the preoperative level 7-14 days after ICH. Conclusion In spontaneously hypertensive rats, the blood pressure regulatory mechanisms following acute ICH can be closely associated with stress-induced activation of the sympathoadrenomedullary system and rennin-angiotensin system.

OBJECTIVEThe pathophysiology of beta-thalassemia is the imbalance of the alpha and non-alpha globin chain which leads to a series of clinical symptoms of hemolytic anemia. Scientists continuously try to explore gene-activated drugs to increase the level of non-alpha globin chain or decrease the level of alpha globin chain in the treatment of beta-thalassemia. To probe into the effects on globin-gene expression of meisoindigo (Me) in cultured erythroid cells derived from peripheral blood, so as to provide the theoretical basis for applying Me in the treatment of beta-thalassemia.

METHODSBy using the two-step liquid culture of erythroid progenitor cells and reverse transcription polymerase chain reaction (RT-PCR), and by using alpha mRNA as an inner control, the level of gamma mRNA and beta mRNA in cultured erythroid cells derived from peripheral blood of 11 patients with severe beta-thalassemia and 6 normal volunteers were measured under the effect of different concentration (2.5 micro mol/L, 5 micro mol/L and 10 micro mol/L) of Me.

RESULTS(1) No statistic significance was found in the ratio of beta/alpha mRNA by Me in cultured cells from both normal individuals and beta-thalassemia. (2) Me can significantly increase the ratio of gamma/alpha mRNA and (beta + gamma)/alpha mRNA (that is non-alpha/alpha mRNA) in cultured cells from normal individuals and beta-thalassemia. The ratio of gamma/alpha mRNA was increased 0.31 - 0.45 times and the ratio of non-alpha mRNA/alpha mRNA increased 0.21 - 0.32 times in Me induced cells from normal individuals. No significant result was observed among the different concentrations of Me (2.5 micro mol/L, 5 micro mol/L and 10 micro mol/L) in normal individuals. With the increasing of Me concentrations, the ratios of gamma/alpha mRNA and alpha/alpha mRNA were increased in cultured cells from beta-thalassemia. The ratio of gamma/alpha mRNA was increased 0.33 - 1.17 times and the ratio of non-alpha/alpha mRNA increased 0.25 - 0.89 times in Me induced cells from beta-thalassemia. There was no significant difference between the concentrations of 2.5 micro mol/L and 5 micro mol/L concentration in beta-thalassemia. However, there was significant difference between the concentrations of 10 micro mol/L and the concentrations of 2.5 micro mol/L and 5 micro mol/L in beta-thalassemia. (3) The increase of the ratio of gamma/alpha mRNA and non-alpha/alpha mRNA in beta-thalassemia was higher than that in normal individual with induction by Me with a higher concentration (10 micro mol/L).

CONCLUSIONMe can raise the ratio of gamma/alpha mRNA and non-alpha/alpha mRNA in cultured erythroid cells derived from peripheral blood of both normal individual and beta-thalassemia in the level of transcription, which can improve the imbalance of the alpha and non-alpha globin chain. So Me has a latent value in the therapy of beta-thalassemia.

Cells, Cultured ; Child ; Child, Preschool ; Erythroid Precursor Cells ; drug effects ; metabolism ; Female ; Gene Expression ; drug effects ; Globins ; genetics ; Humans ; Indoles ; pharmacology ; Infant ; Male ; RNA, Messenger ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the effect of long-term power frequency electromagnetic field (50 Hz) exposure on the proliferation and apoptosis of human lens epithelial cells (SRA01/04 cells).

METHODSSRA01/04 cells in the exponential growth phase were exposed or sham-exposed to power frequency electromagnetic field (50 Hz, 2.3 mT) for 2 hours per day, 5 days every week. After 11 weeks of exposure, the cells were collected; the cell morphology was observed under a microscope, the cell viability was measured by MTT assay, the cell cycle and apoptosis were examined by flow cytometry, and the protein expression levels of cyclin D and proliferating cell nuclear antigen (PCNA) were determined by western blot.

RESULTSCompared with the sham-exposed SRA01/04 cells, most exposed cells became rounded and more stereoscopic, and heterochromatin gathered near the nuclear membrane in some exposed cells. The MTT assay showed that the viability of exposed cells was significantly increased compared with that of the sham-exposed cells (P < 0.05). Long-term power frequency electromagnetic field exposure led to significantly increased number of cells in S phase (P < 0.05), and the proliferation index was significantly higher in the exposed cells than in the sham-exposed cells (P < 0.05). There was no significant difference in apoptotic rate between the exposed cells and sham-exposed cells (P > 0.05). The exposed cells had significantly higher protein expression levels of cyclin D and PCNA than the sham-exposed cells (P < 0.05).

CONCLUSIONLong-term power frequency electromagnetic field exposure can promote cellular proliferation and change cell cycle in SRA01/04 cells, but it has no marked effect on the apoptosis of SRA01/04 cells.

Apoptosis ; Cell Line ; Cell Proliferation ; Cyclin D1 ; metabolism ; Electromagnetic Fields ; adverse effects ; Environmental Exposure ; adverse effects ; Epithelial Cells ; cytology ; Humans ; Lens, Crystalline ; cytology ; Proliferating Cell Nuclear Antigen ; metabolism