Acetic Acid ; analysis ; Chromatography, Ion Exchange ; Workplace

Objective Investigate the Asi-antidiarrheal capsule's effect on gastrointestinal horrnones in blood plasma of thyroid hormone-induced diarrheic(Abbreviation:Hyperthyroid Diarrhea.,D)rats.Methods One hundred and twenty SD male rats about 8 weeks old were randomly divided according to their constitution into control group of 10 rats and thyroid hormone-induced diarrheic group of 110 rats.The control group rats welle hvaged with isotonic Na chloride 1 ml/d.Thyroid tablets were made with isotonic Na chloride into 40 g/L susnl. The such solution with 1 ml/d was intragastrically administered to each rat in thyroid hormone-induced diardleic group.After three weeks,blood was sampled from vena caudalis of each rat.FT4 were then detected in blood semm.Fourty-two thyroid hormone-induced diarrheic rats were screened based on FT3 and FT4 level in blood serum, wet stool and body weisht.Fourty thyroid hormone-induced diarrheic rats were stochastically re-divided into 5 groups with 8 in each.The physiological saline with 1 ml/d was given to blank group,1.94 g·-kg-1·d-1 Berberine capsule to positive control group,and 0.63,1.26,2.52 g·kg-1·d-1 to low-dose,moderate-dose and high-dose groups respectivelv. Intragastric administration of each group continued for 7 days.Venous blood was centrifuged before and after administration and underwent radioimmunoassay to observe the effect of Ashi-antidiarrheal capsule on motillty (MTL),gastrin(GAS),somatostatin(SS),vasoactive intestinal polypeptide(VlP)in blood plasma of thyroid homlone- induced diarrheic rats.Results①Weight of thyroid hormone-induced diarrheic rats decreased[(344.0±12.9)g], FT3[(4,58 ±0.70)mol/L]and FT4[(23.44±4.40)mol/L]increased,and weight of wet stool[(17.4±3.2)g] increased.Compared to control group[(386.0±1.8)g,(2.08±0.10)mol/L,(10.18±2.00)mol/L,(9.1±0.6)g], there was a statistical significance(t=6.85,9.80,7.66,7.18,P<0.01).②After treatment,high-dose Ashi-antidiarrheal group[(80.54 ±3.80)ng/L]and positive control group[(90.63 ±9.99)ng/L]blood plasma MTL, compared to pre-therapy[(204.27±17.69),(187.79±13.32)ng/L]was decreased,there wag a statistical significance (t=8.60,4.57,P<0.01)③GAS contentshad respectively decreased comparedtopre-therapy[(192.75±11.80), (193.09±3.81),(190.60±9.31),(196.33±18.13)ng/L]in positive control group[(56.06 ±6.36)ng/L],low- dose group[(90.88±4.18)ng/L],midst-dose group[(75.64±7.09)ng/L]and hish-dose group[(44.32±3.72) ng/L],except for blank group.There Wag a statistical significance(t=15.27,7.62,13.43,13.22,all P<0.01).The intm-group difference of MTL,GAS and VIP level had statistic signifieances before and after the treatment(F= 166.68,1503.53,216.68,P<0.01).Conclusion Ashi-antidiarrheal capsule Can significantly lower the level of MTL and GAS in blood plasma。And raise the level of VIP.

Objective To study the effect of Asi-antidiarrheal capsule on gastrointestinal goblet cell of thyroid hormone-induced diarrhea.Methods Total of 120 SD male rats aged about 8 weeks were randomly divided into 2 groups:control group(10 rats)and thyroid hormone-induced diarrheic group(110 rats).Rats in control group were lavaged with normal saline 1 ml/d.Thyroid tablets were partly desolved into normal saline forming a 40 mg/ml suspension.Rats in thyroid hormone-induced diarrheic group were given the thyroid suspension 1 ml/d to make thyroid hormone-induced diarrheic model.Serum FT3 and FT4 were tested.Fourty thyroid hormone-induced diarrheic rats were screened out according to serum FT3 and FT4 levels,body weight and wet stool.The fourty rats were randomly divided into 5 groups,8 rats in each group:positive control group,berberine group,low-dose,mediandose and high-dose groups.Normal saline of 1 ml/d was admnistered to diarrhea control group,1.94 g·kg-1·d-1 Berberine capsule was given to positive control group,and 0.63,1.26,2.52 g·kg-1·d-1 Asi-antidiarrheal capsule to low-dose,mediandose and high-dose groups,respectively.After sever days treatment,rats are executed.Duodenum,jejunum,ileum and colon were dissected,respectively.Histology observation and cell counting were carried out under light micmscopo on HE coloration.Cell counting unit was defined as:cell/high power field of vision (cells/hpf).Results In jejunum,the number of goblet cells in berberine group,mediandose group and high-dose group[(15.32±2.53),(20.24±1.24),(14.98±1.10)cells/hpf,respectively],were all lower than that of the diarrhea control group[(25.73±4.55)cells/hpf,all P<0.05]with an exception of low-dose group[(23.98±2.28)cells/hpf].The numbers of goblet cells in berberine control group,low-dose group,mediandose group and highdose group[(18.29±1.33),(20.61±2.12),(19.38±2.01),(16.34±1.55)cells/hpf,respectively]were all less than that of the control group[(23.36±3.10)cells/hpf,all P<0.05].The numbers of goblet cells of diarrhea control group and high-dose group were obviously lower than that of the low-dose group(all P<0.05)in jejunum and colon.The numbers of goblet cells of Duodenum and ileum were not significantly different between groups(F=2.81,2.67,all P>0.05).The numbers of goblet cells in the diarrhea control group increased markedly observed under microscope,but decreased following therapeutic treatment.Conclusions The numbers of goblet cells from jejunum and colon in thyroid hormone-induced diarrheic rats are increased significantly.Asi-antidiarrheal capsule can remarkably decrease the number of goblet cells in jejunum and colon,and reduce mucus secretion.

AIMTo study the feasibility of long-term potentiation(LTP) recording in the CA1 area of the rat in vivo with electrodes-binding technique.

METHODSAnesthetizing Wistar rats with urethane and fixing the animal on the stereotaxic device for acute surgery; implanting cannula into lateral cerebral ventricle; inserting self-made bound stimulating/recording electrodes into hippocampal CA1 area; recording basal field excitatory postsynaptic potential (fEPSP) and tetanus-induced long term potentiation (LTP).

RESULTSfEPSPs were reliably induced by using the stimulating/recording electrodes-binding technique, and the appearance rate of fEPSP was nearly 100%; basal fEPSP recording was very stable, lasting for long time enough to finish all experiment; high frequency stimulation (HFS) successfully induced LTP, which maintained more than three hours, the inductivity is about 67%; paired-pulse facilitation (PPF) recording was also stable; intracerebroventricular (i c v) injection of amyloid beta suppressed HFSinduced LTP evidently.

CONCLUSIONThe electrodes-binding technique for recording hippocampal LTP in vivo is quite simple and convenient. The experimental resource can be saved, and the rates of fEPSP appearance and LTP induction are kept high. Therefore, it is promising for this technique to be one electrophysiological auxiliary method in the research of learning and memory.

Animals ; Electric Stimulation ; methods ; Electrodes ; Excitatory Postsynaptic Potentials ; physiology ; Feasibility Studies ; Hippocampus ; physiology ; Long-Term Potentiation ; physiology ; Male ; Rats ; Rats, Wistar

Although the impairing effects of beta-amyloid (Aβ) protein on synaptic plasticity and cognitive function have been widely reported, the mechanisms underlying the neurotoxicity of Aβ are still not well known. The present study observed the effects of intracerebroventricular (i.c.v.) injection of both Aβ(23-35) and genistein (a specific tyrosine kinase inhibitor at high concentration) on the hippocampal long-term potentiation (LTP) in the CA1 region, and investigated its possible protein tyrosine kinase (PTK) mechanism. Male Wistar rats were surgically prepared for acute LTP recordings in vivo. Two parallel bond electrodes for stimulating and recording were simultaneously inserted into the right hippocampus of rats. The field excitatory postsynaptic potentials (fEPSPs), paired-pulse facilitation (PPF) and high-frequency stimuli (HFS)-induced LTP were recorded by delivering test stimuli, paired pulses and HFS to the Schaffer-collateral/commissural pathway. The results showed that: (1) i.c.v. injection of Aβ(23-35) did not affect the baseline synaptic transmission, but significantly suppressed the HFS-induced LTP, with a decreased average amplitude of fEPSPs [(129.2+/-6.7)% in 10 nmol Aβ(23-35) group; (110.6+/-8.6)% in 20 nmol Aβ(23-35) group; P<0.01] at 1 h post-HFS when compared to that in the control group [(163.1+/-8.1)%]; (2) Similarly, i.c.v. injection of genistein (200 nmol) did not change the basic synaptic transmission, but significantly suppressed HFS-induced LTP, with the similar average amplitude of fEPSPs [(114.0+/-7.2)%] at 1 h post-HFS to that in 20 nmol Aβ(23-35) group; (3) Co-application of Aβ(23-35) (20 nmol) and genistein (200 nmol) caused no additive suppression of LTP, and the average amplitude of fEPSPs was (113.0+/-8.8)% at 1 h post-HFS, showing no significant difference when compared with that in Aβ(23-35) or genistein alone groups (P>0.05); (4) There was no significant change in the PPF following genistein and Aβ(23-35) alone or co-injection (P>0.05). These experimental results indicate that i.c.v. injection of Aβ(23-35) can significantly suppress the HFS-induced LTP in the CA1 area of rat hippocampus in vivo, implying that the Aβ deposited in the brain of patients with Alzheimer's disease may impair the function of learning and memory by suppressing the hippocampal LTP. The facts that the extent of inhibition of Aβ(23-35) and genistein on LTP was similar and no further potentiation of the suppression was observed when Aβ(23-35) and genistein were co-applied suggest that PTK is probably involved in the Aβ-induced suppression of hippocampal LTP.
Amyloid beta-Peptides ; pharmacology ; Animals ; CA1 Region, Hippocampal ; drug effects ; enzymology ; Excitatory Postsynaptic Potentials ; Genistein ; pharmacology ; Long-Term Potentiation ; Male ; Neuronal Plasticity ; Peptide Fragments ; pharmacology ; Protein Kinase Inhibitors ; pharmacology ; Protein-Tyrosine Kinases ; metabolism ; Rats ; Rats, Wistar ; Synaptic Transmission

AIMTo establish separation methods of five sulfonamides by using capillary high performance liquid chromatography(mu-HPLC) and electrochromatography. The effect of mobile phase varies such as methanol content, pH, buffer solution concentration and voltage on their chromatographic behavior and electroosmesis flow was investigated. Capillary electrochromatography (CEC) was compared with mu-HPLC at the same condition.

METHODSStationary phase was ODS, mobile phase was methanol and 2 mmol.L-1 H3PO4 buffer solution (pH 3.0-7.0), voltage was 0- -15 kV, flow rate was 10 microL.min-1, pressure was approximately 70 MPa and UV detection wavelength was 254 nm.

RESULTSSeparations on base line have been respectively accomplished for five sulfonamides by mu-HPLC with mobile phase of methanol-2 mmol.L-1 H3PO4 buffer solution (30:70) at pH 5.0 in 67 min, and CEC with the same mobile phase at -5 kV voltage in 25 min.

CONCLUSIONElectroosmesis flow of CEC decreased with the increase in methanol content, buffer solution concentration, increased with the increase in voltage and increase slightly with the increase in pH of mobile phase. Retention values (k) of solutes to be examined decreased with increasing methanol content of mobile phase in mu-HPLC and CEC. Retention values (k) of solutes increased slightly with increasing buffer solution concentration, decreased with increasing voltage in CEC. Trimethoprim(TMP) decreased obviously with increasing voltage in CEC. The effect of pH of mobile phase on retention values (k) was more complex. Five sulfonamides were separated at the same mobile phase condition by mu-HPLC and CEC. And separation speed of CEC was much faster than that of mu-HPLC. CEC was very fit for rapid separation of sulfonamides.

Anti-Infective Agents ; isolation & purification ; Buffers ; Chromatography, High Pressure Liquid ; methods ; Chromatography, Micellar Electrokinetic Capillary ; methods ; Hydrogen-Ion Concentration ; Sulfonamides ; isolation & purification ; Trimethoprim ; isolation & purification

OBJECTIVETo explore the application value of detection of Hepcidin together with indicator of iron overload on clinical diagnosis and treatment of MDS with iron overload by measuring Hepcidin and iron load indices of transfusion dependent myelodysplastic syndrome (MDS) patients.

METHODSEnzyme-linked immunosorbent assay (ELISA), radioimmunoassay and colorimetry were used to determine the Hepcidin, serum ferritin (SF) and serum iron (SI) levels of 106 serum samples from 68 cases of transfusion dependent MDS patients, 30 serum samples of MDS patients without transfusion and 60 serum samples of controls.

RESULTSFor MDS group, Hepcidin level in blood transfusion < 9 U subgroup was significantly higher than that in control group \[(583 ± 50) µg/L vs (175 ± 35) µg/L\] and there was a strong positive correlation between Hepcidin levels and SF (r = 0.976), but no correlation between Hepcidin and SI (r = 0.284); Both Hepcidin and SF level in transfusion 9 ∼ 24 U subgroup was significantly higher than those in control group \[(665 ± 80) µg/L vs (175 ± 35) µg/L; (1445 ± 275) µg/L vs (112 ± 26)µg/L\]; whereas for SI level, there was no difference between transfusion 9 ∼ 24 U subgroup and the control group. Hepcidin did not correlate with SF or SI; For blood transfusion > 24 U group, all of Hepcidin, SF and SI levels were higher than those in control groups \[(703 ± 64) µg/L vs (175 ± 35) µg/L; (2587 ± 352) µg/L vs (112 ± 26)µg/L; (20 ± 4) µg/L vs (14 ± 4) µmol/L\], Hepcidin negatively correlated with SF and SI (r = -0.536; r = -0.456). Hepcidin levels of RARS patients were significantly lower than RAEB patients \[(260 ± 40) µg/L vs (442 ± 51) µg/L\], and there was no significant difference between RARS group and control group regardless of the number of blood transfusion.

CONCLUSIONBoth Hepcidin and SF levels in MDS patients regardless of transfusion dependent or not, or the number of blood transfused were higher than those of normal controls, the increase of Hepcidin can not synchronize with the increase of SF level due to the increased blood transfusion, when blood transfusion > 24 U, Hepcidin level showed a negative relationship with SF and SI, reflecting the decreased ability of Hepcidin to inhibit body iron absorption during the increase of blood transfusion, which finally would lead to iron overload. We can predict the occurrence of iron overload in transfusion dependent MDS patients by dynamic monitoring concentration of Hepcidin.

Adult ; Aged ; Aged, 80 and over ; Antimicrobial Cationic Peptides ; blood ; Blood Transfusion ; Female ; Ferritins ; blood ; Hepcidins ; Humans ; Iron ; blood ; Iron Overload ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; therapy

Adult ; Chylous Ascites ; therapy ; Female ; Fibrin Tissue Adhesive ; therapeutic use ; Humans ; Middle Aged ; Parenteral Nutrition, Total ; Somatostatin ; therapeutic use

An alkaline catalase has been purified and characterized from a slightly halophilic and alkaliphilic bacterium Bacillus sp. F26. The purification was performed with a four step procedure consisting of ammonium sulfate precipitation, ion exchange, gel filtration and hydrophobic interaction chromatography, and finally achieved a 58.5-fold-purifying over the crude extract. The purified catalase was composed of two identical subunits with a native molecular mass of 140 kD. The native enzyme showed the typical Soret band appearing at 408 nm. The pyridine hemochrome spectrum indicated the presence of protoheme IX as the prosthetic group. The apparent Km value for enzyme activity on H2O2 was calculated to be 32.5 mmol/L. The activity of this catalase was not reduced by dithionite but was strongly inhibited by cyanide, azide, and 3-amino-1,2,4-triazole (the specific inhibitor of monofunctional catalase). No peroxidase activity of this enzyme was detected when using o-dianisidine, diaminobenzidine (DAB) and p-phenylenediamine as electron donor. Moreover, the N-terminal sequence of this catalase exhibited substantial similarity to the monofunctional catalase subgroup rather than catalase-peroxidase or Mn-catalase one. Therefore, we characterize the purified catalase as a monofunctional catalase. Besides, this monofunctional catalase was thermosensitive and its activity exhibited pH-independent over pH 5-9 but showed a sharp maximum at pH 11. An activity half-life of approximately 49 h was measured when the enzyme was incubated at 20 degrees C and pH 11. To our knowledge, pH 11 is the most alkaline condition for optimum catalysis and enzyme stability among the catalases reported up to now. Furthermore, this monofunctional catalase also showed excellent halo-alkali-stability with a half-life of approximately 90 h at 0.5 mol/L NaCl and pH 10.5. On the other hand, so far as we know, the characterized catalase is the first dimeric monofunctional catalase from alkaliphiles and is also the first monofunctional catalase derived from a natural soda lake, which could partially reflect the oxidative stress response in the corresponding environment.
Bacillus ; enzymology ; Bacterial Proteins ; chemistry ; isolation & purification ; Catalase ; chemistry ; isolation & purification ; Enzyme Stability ; Hydrogen-Ion Concentration ; Temperature

The amyloid β-protein (Aβ)-induced disturbance of intracellular calcium homeostasis has been regarded as the final route whereby Aβ insults neurons. However, the mechanism of Aβ-induced Ca(2+) overloading is still unclear so far. Especially, it remains to be clarified whether nicotinic acetylcholine receptors (nAChRs) are involved in the Aβ-induced elevation of intracellular calcium concentration ([Ca(2+)](i)). In the present study, we observed the effects of Aβ fragments 25-35 (Aβ(25-35)) and 31-35 (Aβ(31-35)) on [Ca(2+)](i) in primary cultured rat cortical neurons using laser-scanning confocal calcium imaging technique, and investigated its probable cholinergic mechanism. The results showed that: (1) Both Aβ(25-35) and Aβ(31-35) induced similar and significant [Ca(2+)](i) elevation in a concentration-dependent manner, and no statistical difference was found between the effects of both peptides; (2) The reverse peptide of Aβ(31-35), i.e. Aβ(35-31), had no effect on [Ca(2+)](i) elevation; (3) Mecamylamine (MCA), a non-specific nAChRs antagonist, significantly and dose-dependently blocked the [Ca(2+)](i) elevation induced by Aβ(25-35) or Aβ(31-35) (4) Dihydro-β-erythroidine (D-β-E), a specific α4β2 subtype nAChRs antagonist, also significantly inhibited the [Ca(2+)](i) elevation induced by Aβ(25-35) and Aβ(31-35), but the effect was weaker than the effect of MCA at the same concentration. These results indicate that Aβ(31-35) may be a shorter active sequence in full length of Aβ molecule, and the overactivation of nAChRs, including α4β2 subtype, may be, at least partly, responsible for the Aβ-induced elevation of [Ca(2+)](i) in cultured rat cortical neurons. Thus, the present study suggests a new potential target of Aβ in the brain, and provides a new insight into the mechanisms by which Aβ impairs the cognitive function in Alzheimer's disease.
Amyloid beta-Peptides ; chemistry ; Animals ; Calcium ; metabolism ; Cells, Cultured ; Neurons ; metabolism ; Peptide Fragments ; chemistry ; Rats ; Receptors, Nicotinic ; metabolism