With the methods of light microscopic histochemistry and electron-microscopic cytochemistry, we studied the changes of lactate dehydrogenase (LDH) and succinic dehydrogenase (SDH) in the anoxic myocardium of the mice; to which normal saline (INSAM group) or 10％ aqueous extract of Ginsengradix rubra (IGRAM group) was injected before the anoxic experiment. The results were as follows:Ⅰ. The Light Microscopic Study:1. In the myocardial sections of the INSAM group, the reaction of SDH of myofibrillae are variegated and the fibrillae vary in thickness. Granular mitochondria are clearly stained and arranged in alignment in the intermyofibrillar spaces. The intercalated discs fail to be stained and their spaces widen. In the IGRAM group, the reactions of SDH are similar to that of the normal control. The enzyme reaction product in the myofibrillae appear as homogeneous diffuse blue precipitate of formazan, the cross striations of myofibrillae are clear and strings of mitochondria are visible.2. The reactions of LDH in the INSAM group reveals that myofibrillar staining is also lost in a length of myofibrilla, strings of mitochondria are clearly stained. The reaction in the IGRAM group is similar to that of the normal control. In the myofibrillae there is a homogenous continuous violet precipitate of formazan.Ⅱ. The Electron-Microscopic Study:1. The reaction of SDH in the INSAM group shows that the enzyme activity in the mitochondrial inner membrane and the cristae is stronger than that of the normal control. The increased activity is represented by the increase of electronic density or darkening and thickening of the membranes. However, the activity in the IGRAM group is simillar to that of the normal control but weaker in reaction.2. The reaction of LDH in the INSAM group reveals that the enzyme activity exhibited by membrane darkening is obviously stronger than that of the normal control but in the IGRAM group it shows clearly a weaker reaction.The results mentioned above suggest that Ginsengradix rubra might alleviate the metabolic disturbance caused by myocardial anoxia.

Humans ; Medicine, Chinese Traditional ; Nephrotic Syndrome ; diagnosis ; therapy

Objective To investigate the changes in Bcl-2-associated athanogene 1(BAG-1)expression,and the mechanism of nuclear translocation in rat alveolar macrophages(AMs)induced by lipopolysaccharide(LPS)and dexamethasone(Dex).Methods Primary culture AMs treated by LPS and Dex were divided randomly into three groups:6h group,2h group and 24h group.The BAG-1 expression in AMs was detected with Western blot.The interactions between BAG-1 and glucocorticoid receptor(GR)were detected with immune co-precipitation.The changes in GR expression in nuclear protein were evaluated with Western blotting after transfection of RNA interference recombinant plasmids(named psilencer 3.1-GR)targeting to GR gene.Results The expression of BAG-1L in total protein increased,and that of BAG-1S showed no changes.Only BAG-1L,with no BAG-1S,was detected in nuclear protein,and its expression increased gradually in 24h.Interaction between BAG-1L and GR was found in nucleolus after treatment.After transfection of plasmids psilencer 3.1-GR,the BAG-1L expression in nuclear protein decreased significantly compared with that of non-transfection group(P

Objective To construct cDNA library of alveolar macrophages after lipopolysaccharide (LPS) and dexamethasone (Dex) treatment for exploring the protein molecule interactive with glucocorticoid receptor (GR) in condition of inflammation. Methods After the cultured AMs were treated with LPS (10mg/ml) and Dex (10 -5 mol/L) for 4h, total RNA was extracted from AMs, then the cDNA was synthesized from total RNA of AMs and amplified using primers SMARTⅢ TM and CDSⅢoligo (dT) as the base of recombination. The purified PCR products as well as the linearized plasmid pGADT7-Rec were co-transformed into the competent yeast AH109. They were recombined by yeast homologous recombinase in the yeast cells and became the active cyclic plasmid. The transformed yeasts grew in the SD/-Leu plates. All the growing clones were harvested and then constituted the cDNA library. Furthermore, the bait pGBKT7-rGR transformed the yeast AH109 of library was constructed, and the protein molecule interactive with GR was screened. Results cDNA library of AMs was constructed with high multiplication and good capacity. 1.07?106 recombinants were obtained from the cDNA library. The amplified PCR fragments were between 0.3-1.5kb in size. One true positive clone, obtained by screening the cDNA library, was confirmed to be BAG-1 by sequencing and BLAST. Conclusion The yeast two-hybrid cDNA library of AMs was successfully constructed by Clontech SMART method; GR can interact with BAG-1 in yeast cells.

Objective To measure 95% effective doses (ED95) of propofol and etomidate by using up-down intravenous administration method, and compare the potency ratio and the anesthesia duration of them. Methods Twenty eight male SD rats were divided into two groups randomly: the propofol group and the etomidate group. Loss of righting reflex was regarded as the judgment index of unconsciousness. The dose-response curve was made according to the formula of Y=Ymin+(Ymax-Ymin)/ [1+10log(ED50-X)× m]. Values of ED95 of propofol and etomidate were calculated, and the anesthesia duration periods after administration of the two equivalent dose drugs were measured respectively. Results The values of ED95 were 9 mg/kg and 1.5 mg/kg for propofol and etomidate. The ED95 ratio for propofol and etomidate was 6. There was a significant difference in anesthesia duration between propofol group (465.6±18.5)s and etomidate group (233.7±9.3)s (P<0.05). Conclusion The anesthesia duration of propofol is longer than that of etomidate, taking the ED95 as equivalent dose.

To develop a method for the determination of residual solvents inα-ketophenylalanine calcium by capillary gas chromatography. Methods:The residual solvents were separated on a DB-624(30 m × 0. 32 mm, 0. 25 μm) capillary chromato-graphic column with temperature programming. The column temperature was maintained at 40℃ for 1 min,and then raised to 180℃at a rate of 10℃·min-1 and maintained for 2 min. N2 was used as the carrier gas, and FID was used as the detector with temperature of 250 ℃. The injector temperature was 200 ℃ and the split ratio was 10∶1, and direct injection was adopted. Methanol, ethanol, ethyl acetate and tetrahydrofuran in α-ketoleucine calcium were detected using an external standard method. Results:The four solvents were separated completely. There was a good linear relationship between the peak area and the concentration of each solvent ( r=0. 997 2-0. 999 5). The average recovery of the four solvents was 95. 47%-100. 26%(RSD≤4. 7%, n=9). Conclusion:The method is rap-id, simple, accurate and sensitive, and can be used in the determination of residual solvents in α-ketophenylalanine calcium.

Objective To evaluate the effect of intrathecal (IT) NR2B antisense oligonucleotide (aNR2B) on cognitive function in morphine-dependent rats.Methods Male SD rats weighing 230-270 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital 60 mg/kg.IT catheter was placed at L3-4 interspace according to the technique described by Yang. Thirty rats in which IT catheter was successfully placed without any complication were randomly divided into 3 groups(n=10 each):control group (group C), morphine dependence group (group MD) and group aNR2B.Morphine dependence was induced in group MD and aNR2B by increasing doses of morphine for 6 days. The initial dose of morphine was 10mg/kg injected subcutaneously (SC) twice a day and was increased by 10 mg/kg.every other day.The final dose was 50mg/kg. Then morphine 30 mg/kg was administered SC once a day for 4 weeks. aNR2B 15 nmol was administered IT at 30 min before SC morphine every day in group aNR2B.In control group normal saline was administered instead of morphine. Morris water maze was used to assess the cognitive function at 0 (T0, baseline),1 and 3 weeks of morphine administration (T1,T2).The escape latency and the number of times the animals crossing the plateform were recorded. The animals were killed after the test and the hippocampus was isolated for determination of choline acetytransferase(ChAT)expression.Results There was no significant difference in the baseline escape latency and the baseline number of times the animals crossing the plateform at T0 among the 3 groups. The escape latency was significantly prolonged and the number of times the animals crossing the plateform decreased at T1 and T2 as compared with the baseline at T0 in group MD.The ChAT expression was significantly down-regulated in group MD as compared with control group. IT aNR2B significantly ameliorated cognitive dysfunction at T1 and T2 and increased ChAT expression in group aNR2B compared with group MD.Conclusion IT NR2B antisense oligonucleotide can attenuate cognitive dysfunction through up-regulation of ChAT expression in hippocampus in morphine-dependent rats.

The epithelial cells of human prostatic hyperplasia had been studied cytochemically for 5'-nucleotidase and thiamine pyrophosphatase activity with light and electron microscopy. The 5'-nucleotidase (5'-Nase) activities were present in the epithelial cells at lysosomes, Golgi vacuoles, vesicles, secretory vacuoles and free surface of plasmic membrane, the secretion material within the acinar lumina of the prostate gland also showed 5'-nucleotidase reaction product. The activity of 5'-nucleotidase has not markedly changed by the actinomyces globispores treatment, but slightly decreased after estrogen treatment. It was no enzyme reaction after treated by testectomy.Activities of thiamine pyrophosphatase (TPPase) were restricted to the Golgi area, and there were heavy deposits of reaction products in concave face of Golgi saccule. The activities of TPPase were not markedly changed by estrogen and actinomyces globispores treatments but none of enzyme activities could be seen after testectomy.

Objective: To investigate the associations of the mutation status of epidermal growth factor receptor (EGFR) with the factors including gender, age, smoking status, family history and histological type in patients with non-small cell lung cancer (NSCLC). Methods: From February 2008 to May 2012, 538 consecutive NSCLC patients went through the clinical testing for EGFR mutations in Shanghai Pulmonary Hospital were included in this study. Amplification refractory mutation system (ARMS) method was used to detect EGFR mutation status. The relationships between EGFR mutation and potential related factors were evaluated by using Logistic regression model. Results: The activating mutation of EGFR was found in 220 patients. EGFR mutation was more frequently detected in females (P < 0.001), patients with adenocarcinoma (P < 0.001), never-smokers (P < 0.001) and patients with family history of cancer (P = 0.031). Multivariate analysis revealed that pathologic type [odds ratio (OR) = 0.035)], smoking status (OR = 0.005) and family history of cancer (OR = 0.027) were independent predictive factors of EGFR mutation. Conclusion: The factors of pathologic type, smoking status and family history of cancer are independent predictive factors of EGFR mutation. Copyright © 2014 by TUMOR.

By means of pharmacohistochemical regimen method and image analysis system,the age-related changes of acetylcholinesterase activity of the substantia nigra neu-rons and the number and size(section area)of the AChE positive neurons in ratswere investigated quantitatively.The results showed that the number of AChE pos-itive neurons in the substantia nigra of the old rats decreased markedly,the rateand quantity of AChE synthesis in the perikaryon remarkable declined.The size(section area)of the AChE positive neurons also reduced with aging.These changeswith aging have never been studied in human or usual experimental animals,so thatsome new parameters were provided for the research field in experimental geronto-logy.In human and animal,the degeneration of the neurons in substantia nigra cau-sed by aging would disturb the balance between dopaminergic and acetylcholinergicsystem and hence interfere with the normal coordination of movement.