OBJECTIVETo economically obtain the Abeta peptide for Alzheimer's disease (AD) research by expressing the Abeta peptide fused with the maltose binding protein (MBP) possessing high solubility in E.coli.

METHODSThe cDNA-coding sequence of Abeta peptide was modified by the addition of a BamH I site at the 5' end and a Hind III site at the 3' end using PCR. The modified sequence was ligated into the maltose-binding protein (MBP) fusion expression vector pMAL-c2 containing an thrombin cleavage site, which was transformed into competent E.coli DH5alpha cells. After identification of the single clones by PCR and DNA sequencing, the recombinant plasmid was transformed into E.coli TB1 and induced to express MBP-Abeta fusion protein. The expressed fusion protein was purified using amylose resin column and identified by SDS-PAGE and Western blotting.

RESULTSThe result of DNA sequencing verified the consistency between the inserted sequence and Abeta (1-42) sequence. SDS-PAGE electrophoresis showed that MBP-Abeta fusion protein was highly expressed in E.coli TB1, and Western blotting demonstrated that the purified fusion protein and the separated Abeta peptide could be recognized by specific anti-Abeta (22-35) antibody.

CONCLUSIONMBP-Abeta fusion protein highly expressed in E. coli TB1 cells with enhanced solubility and the separated Abeta peptide with good immunogenicity obtained may lend support to AD research.

Alzheimer Disease ; genetics ; Amyloid beta-Peptides ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Maltose-Binding Proteins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

Aged ; Biopsy, Needle ; methods ; Coal Mining ; Humans ; Lung ; pathology ; Lung Neoplasms ; complications ; diagnosis ; Middle Aged ; Pneumoconiosis ; complications

Adult ; Aged ; Cell Cycle Proteins ; analysis ; Coal Mining ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Immunohistochemistry ; Lung ; chemistry ; pathology ; Male ; Middle Aged ; Occupational Diseases ; metabolism ; Pneumoconiosis ; metabolism ; Tumor Suppressor Proteins ; analysis

OBJECTIVETo study the protective effects of salidroside on injury induced by hypoxia/hypoglycemia in cultured SH-SY5Y cell.

METHODApoptosis and intracellular free calcium concentration ([Ca2+]i) were measured by flow cytometry, morphological changes and neuronal necrosis were observed with fluorescence microscope, and the lactic dehydrogenate (LDH) release was measured by LDH kits.

RESULTHypoxia/hypoglycemia cultures for 2 hours induced neuronal apoptosis and necrosis. They were 18.59% (P < 0.01) and 4.94% (P < 0.01) respectively. There were morphological changes including chromatin condensation, nuclear fragmentation and formed apoptotic bodies after exposed to hypoxia/hypoglycemia for 2, 4, 6, 12 hours. After 2 hours of hypoxia/hypoglycemia, neuronal [Ca2+]i and the release of LDH were significantly increased. They were 8.46 nmol/L (P < 0.01) and 16.59% (P < 0.01) respectively. The effects were enhanced with the extending time of hypoxia/hypoglycemia. Salidroside might have significantly decreased the percentage of neuronal apoptosis and necrosis, reduced neuronal [Ca2+]i and the release of LDH. The effects of salidroside were strengthened with the increasing of Salidroside dosage.

CONCLUSIONSalidroside has effect of anti-neuronal apoptosis. This effect might be related to its function of decreasing intracellular free calcium concentration.

Apoptosis ; drug effects ; Calcium ; metabolism ; Cell Hypoxia ; drug effects ; Glucosides ; isolation & purification ; pharmacology ; Humans ; Hypoglycemia ; metabolism ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Phenols ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rhodiola ; chemistry

Objective To evaluate the characteristics of myocardial 18F-FDG imaging in pilots with regular aerobic exercise training. Methods Twenty seven healthy male pilots with regular aerobic exercise training were included in this study. The subjects were divided into fasting(n=17) or non-fasting group (n=10). Fluorine-18-labeled deoxyglucose and Tc-99m-sestamibi dual-nuclide myocardial imaging were obtained at rest and at target heart rate during bicycle ergometer test. The exercise and rest myocardial perfusion imaging were analyzed for myocardial ischemia presence. The myocardial metabolism imaging was analyzed with the visual semi-quantitative analyses model of seventeen segments. Results The secondary-extreme heart rate (195-age) was achieved in all subjects. There was no myocardial ischemia in all perfusion imaging. In the visual qualitative analyses, four myocardial metabolism imaging failed in the fasting group while one failed in the non-fasting group (P > 0.05 ). In the visual semi-quantitative analyses,myocardial metabolism imaging scores at rest or exercise in all segments were similar between two groups (P>0.05). In the fasting group, the myocardial metabolism imaging scores during exercise were significantly higher than those at rest in 6 segments (P < 0.05 ). In the non-fasting group, the scores of 3 exercise myocardial metabolism imaging were significantly higher than those at rest ( P < 0.05). Conclusion Satisfactory high-quality myocardial metabolism imaging could be obtained at fasting and exercise situations in subjects with regular aerobic exercise.

[Objective]To study the intervention effect of mild hypotherrnia on cerebral edema in rats after cerebral infarction.[Methods]Twelve male SD rats were randomized into control group (n--6) and mild hypotherma treatment group (n=6).The rat models of focal cerebral infarction were established with obstructing the middle cerebral artery of rats by suture emboli.Rats of the mild hypotherma treatment group were treated by mild hypotherm for 3 h after the models were established.MRI was performed at 6 h,and 1,3,5 and 7 d after the success of model making.Cerebral infarction volume,absorption rate of edema,and signal intensity ratios (SIRs) ofTIWl,T2WI,FLAIR and DWI sequencas in the infarction zones,and relative variance ratio of SIR (△SIR) were measured and calculated.[Results] In the mild hypotherma treatment group,the cerebral infarction volumes 3,5 and 7 d after the models were established were significantly decreased as compared with those in the control group (P＜0.05);the absorption rate of edema (the 7th d against the 3th d of infarction) in the the mild hypotherma treatment group was significantly higher than that in the control group (P＜0.05).Three,5 and 7 d after infarction,the SIRs of TIWI were all obviously higher than those in the control group,and the SIRs of T2WI and FLAIR were all significantly lower than those in the control group (P＜0.05).The SIR of DWI in the mild hypotherma treatment group was obviously lower than that in the control group 7 d after infarction (P＜0.05).The △SIRs of T2Wl,FLAIR and DWI in the mild hypotherma treatment group were significantly higher than those in the control group (P＜0.05).[Conclusion] Mild hypothermia has a significant inhibition effect on cerebral edema in rats after cerebral infarction.

The steps for installation and withdrawal of WYD2000 field surgical lamp were introduced.The failures and causes of broken cross-arm connector of WYD2000 field surgical lamp were analyzed.The problems of WYD2000 field surgical lamp in vulnerability to breaking and difficulty in maintenance were solved by designing and manufacturing a special maintenance tool and optimizing the materials and fixing mode of cross-arm connection.References were provided for main-tenance and improvement of WYD2000 field surgical lamp.[Chinese Medical Equipment Journal,2024,45(4):116-118]

Objective To observe the clinical efficacy of self-prescribed Guchong Decoction combined with Ferroids for the treatment of iron deficiency anemia (IDA)patients with unconsolidation of thoroughfare and conception vessels. Methods A total of 70 IDA patients with unconsolidation of thoroughfare and conception vessels were randomized into treatment group and control group,35 cases in each group. The control group was given oral use of Ferroids, and the treatment group was treated with self-prescribed Guchong Decoction combined with Ferroids. One month constituted a treatment course,and both groups were treated for 2 courses. Before and after treatment,we compared the parameters of blood routine test for red blood cells(RBC),hemoglobin (HGB), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), MCH concentration (MCHC),white blood cells(WBC)and platelets,as well as reticulocytes(Ret)and iron metabolism indexes including serum iron(SI),serum ferritin(SF),and total iron-binding capacity(TIBC). The adverse reaction was monitored during the treatment. After treatment,both western medicine clinical efficacy and traditional Chinese medicine (TCM)syndrome clinical efficacy of the two groups were evaluated. A 3-month follow-up was carried out after treatment to evaluate the recurrence rate. Results(1)Drop-out occurred in one case from the treatment group and in 2 cases from the control group because of short-term medication suspension.(2)After treatment for 2 months, western medicine clinical cure rate of the treatment group was 82.4％ and that of the control group was 60.6％, and TCM syndrome clinical cure rate of the treatment group was 94.1％ and that of the control group was 51.5％,the difference being significant(P < 0.05 or P < 0.01).(3)The levels of RBC,HGB,Ret, MCV, MCH, MCHC, and WBC of the two groups were obviously increased after treatment (P < 0.01), and PLT level was markedly decreased (P < 0.01)as compared with those before treatment,but the inter-group differences after treatment were insignificant (P > 0.05). (4)After treatment,iron metabolism indexes of SF and SI of the two groups were significantly increased (P < 0.01), and TIBC of the treatment group was significantly decreased (P < 0.01). The treatment group had stronger effect on increasing SF and SI and on decreasing TIBC than the control group (P < 0.05 or P < 0.01). (5)The incidence of adverse reaction and the recurrence rate in the treatment group were lower than those of the control group,the difference being significant (P< 0.01). Conclusion Guchong Decoction combined with Ferroids is effective for the treatment of IDA with unconsolidation of thoroughfare and conception vessels through relieving symptoms rapidly, reducing gastrointestinal adverse reaction,and decreasing recurrent rate.

OBJECTIVEStroke is a complex disorder caused by a combination of genetic and environmental factors. Epidemiological studies have provided evidence of genetic influence on the development of human stroke. However, genetic changes which contribute to the development of stroke are not well known. This study was designed to gain a deep insight into that aspect.

METHODSUsing cold-stimuli plus high-salt intake as environmental risk factors, the authors established a hypertension model in rats, which produced a complication of stroke. Then, they used the suppression subtractive hybridization(SSH) technique to identify the differential genes that specifically expressed in total cerebrum tissue of the rats in stroke group. A comparison was made between two populations, namely the control group and stroke group.

RESULTSBy the use of SSH approach, a total of 576 clones were generated in this study from two subtractive libraries, among them 456 clones were usable and were analyzed. Genes for metabolism transcripts in stroke group were shown to be up-regulated (P<0.01). Mitochondrial transcripts were observed in a high rate of 26.5%.

CONCLUSIONThe findings suggested that mitochondrial genes should induce an increased sensitivity to stroke through the changes of gene expressions. Mitochondrial genes probably play important roles in the causes and effects of stroke.

Animals ; Brain ; metabolism ; pathology ; DNA, Mitochondrial ; genetics ; Gene Expression Regulation ; Male ; Mutation ; Rats ; Rats, Wistar ; Stroke ; etiology ; genetics ; pathology

OBJECTIVETo study the effect of L-alanyl-L-glutamine (Ala-Gln) on the levels of insulin-like growth factor-1 (IGF-1) and IGF-1 receptor (IGF-1R) in the intestinal tissues of low-birth-weight (LBW) newborn rats with hypoxia/reoxygenation-induced intestinal injury.

METHODSPregnant rats were fed with or without smoking. The rats born by those fed without smoking were included in group A; for the rats born by those fed with smoking, normal-birth-weight rats were included in group B, and LBW rats were randomly divided into control group (group C), hypoxia/reoxygenation (H/R) group (group D), and Ala-Gln group (group E). Each group consisted of 24 newborn rats. The rats in groups D and E received H/R treatment twice a day for three consecutive days to establish an intestinal injury model; the rats in group E were intraperitoneally injected with Ala-Gln (10 ml/kg) before daily H/R treatment, while those in groups C and D were given an equal dose of normal saline by intraperitoneal injections. On days 4, 7, and 10 after birth, 8 rats were sacrificed in each group to collect intestinal tissues. The IGF-1 levels in intestinal tissues were measured using ELISA, and IGF-1R levels were measured by immunohistochemistry.

RESULTSThere were no significant differences in IGF-1 and IGF-1R levels between groups A and B at all time points. The levels of IGF-1 and IGF-1R in group C kept increasing, were higher than those in other groups on day 7 (P<0.05), and reached a normal level on day 10, without significant differences compared with those in groups A and B. Group D had significantly lower IGF-1 and IGF-1R levels than group C at all time points (P<0.05). The levels of IGF-1 and IGF-1R in group E were lower than those in group C on days 4 and 7 (P<0.05), but they increased to approximately the levels in group C and were significantly higher than those in group D on day 10.

CONCLUSIONSIntrauterine and postnatal hypoxia may induce intestinal injury in LBW newborn rats, and parenteral administration of high-dose Ala-Gln can reduce hypoxia-induced intestinal injury. Therefore, Ala-Gln has a protective effect against hypoxia-induced intestinal injury.

Animals ; Birth Weight ; Dipeptides ; pharmacology ; Female ; Hypoxia ; metabolism ; Insulin-Like Growth Factor I ; analysis ; Intestines ; chemistry ; Male ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Receptor, IGF Type 1 ; analysis