This study aims to establish a method to determine the serum acetaminophen concentration based on diazo reaction, and apply it in the gastric emptying evaluation. Theoretically, acetaminophen could take hydrolysis reaction in hydrochloric acid solution to produce p-aminophenol, which could then take diazo reaction resulting in a product with special absorption peak at 312 nm. Then the serum acetaminophen concentration and recovery rate were calculated according to the standard curve drawn with absorbance at 312 nm. ICR mice were given a dose of acetaminophen (500 mg x kg(-1)) by gavage and the serum acetaminophen was dynamically measured through the diazo reaction. Besides, ICR mice were subcutaneously injected with the long-acting GLP-1 analog GW002 before the gavage of acetaminophen, and serum acetaminophen concentration was measured as above to study how GW002 could influence the gastric emptying. The data showed acetaminophen ranging from 0 to 160 μg x mL(-1) could take diazo reaction with excellent linear relationship, and the regression equation was y = 0.0181 x +0.0104, R2 = 0.9997. The serum acetaminophen was also measured with good linear relationship (y = 0.0045 x + 0.0462, R = 0.9982) and the recovery rate was 97.4%-116.7%. The serum concentration of acetaminophen reached peak at about 0.5 h after gavage, and then gradually decreased. GW002 could significantly lower the serum acetaminophen concentration and make the area under the concentration-time curve (AUC) decrease by 28.4%. In conclusion, a method for the determination of serum acetaminophen based on the diazo reaction was established with good accuracy and could be used in the evaluation of gastric emptying.
Acetaminophen ; blood ; pharmacokinetics ; Aminophenols ; Animals ; Gastric Emptying ; Mice ; Mice, Inbred ICR

Objective To observe the effect of flexible laryngeal mask (FLMA) in transnaso-sphenoidal microsurgery for pituitary adenoma on airway management and variation of stress response.Methods One hundred patients (71 males, 29 females, aged 18-65 years, BMI 21-28 kg/m2, ASA physical status Ⅰ or Ⅱ) undergoing transnaso-sphenoidal microsurgery for pituitary adenoma were randomly divided into two groups: the FLMA group (group F) and the reinforced endotracheal tube group (group T) using a random number table, 50 cases in each group.The plasma concentration of epinephrine and norepinephrine were measured before anesthesia induction (T0), at the time of inserting the FLMA or reinforced endotracheal tube (T1), 1 min (T2) and 5 min (T3) after insertion.The Berry scores of the preoperative and postoperative airway exposure by branchofiberoscope in group F were assessed.The time of removal of FLMA (endotracheal tube) and the occurrence of choking, laryngeal spasm, sore throat, hoarseness and other adverse reactions were recorded.Results The levels of epinephrine and norepinephrine were were significantly lower at T2 and T3 in group F than those in group T (P<0.05).There was no significant difference in airway Berry scores.The time of extubation was shorter in group F than that in group T [(9±3) min vs (17±6) min] (P<0.05).The incidence of choking (2% vs 22%) and sore throat (4% vs 30%) were significantly lower in group F than those in group T (P<0.05).Conclusion Compared with the reinforced endotracheal tube, FLMA can be applied safely and effectively to transnaso-sphenoidal microsurgery for pituitary adenoma, reduces stress respond associated with anesthesia and post-extubation complications, improves the recovery of patients.

ObjectiveTo understand the characteristics of mutations in BCR-ABL1 kinase domain mutation,these chronic myeloid leukemia (CML) and Ph positive acute lymphoblastic leukemia (ALL)patients who got imatinib treatment had poor effect.MethodsTotally 177 CML patients and 33 Ph( + )ALL patients were selected at Beijing Dao-Pei Hospital from Sep.2007 to Dec.2010.All of them were Chinese patients.Totally 243 bone marrow or peripheral blood specimens were collected from the patients,who had early effect,then resistance emergenced,or for more than 3 months of poor efficacy.Extracted total RNA from the specimens' nuclear cells,reversed transcription to cDNA.Amplified the whole span of BCRABL1 fusion kinase gene by nest PCR (from 242 to 493 amino acid coding sequence),used the type AB3130XL gene sequencing instrument determinate the gene sequence of ABL1 kinase region and then used the Variant Reporter V1.0 software to analyze the results of gene mutations.ResultsThirty-two kinds of different mutations were detected of ABL1 gene mutations,accounting for 34.2％ (83/243 cases).Among them,the T315I was 12％ (10/83),mutation rate was the highest,followed by Y253H was 11％ (9/83),G250E was 7％ (6/83),E255K was 7％ (6/83),M351T was 6％ (5/83),E459K was 5％ (4/83) ;Q252H,D276G,F317L,E355G,F359V,H396R were all 4％ (3/83).Three cases of insertion mutations were found,including 2 cases of 357-358insk,1 case of V304RfsX17.Seven patients had found existence two or more point mutations.The multiple drug resistance mutations might exist in the same leukemia clone.The same individual was not only contain common resistance mutations,but also rare point mutations,insertion mutations.The mutations might be lead to loss of kinase activity.ConclusionsUnder the imatinib drugs pressure,the ABL1 gene mutation in leukemia cells appears randomly,and results in different resistant clones.Different resistant clones can coexist in the same patients in vivo; resistant clones not only contain point mutations,but also contain inserted deletion mutations.

OBJECTIVETo investigate the value of iodine-131 therapy for hyperthyroidism complicated hyperthyroid heart disease(HHD) induced by Graves' disease or Plummer disease.

METHODSTotally 40 HHD cases who were confirmed in our department from 2009 to 2010 were enrolled in this study. All patients received serum thyroid hormones and associated antibodies tests, 12-lead electrocardiogram, and/or thyroid imaging before and after iodine-131 therapy to access the treatment effectiveness.

RESULTSAmong 31 patients with HHD due to Graves' disease and 9 due to Plummer disease, iodine-131 treatment resulted in euthyroidism in 15 and 5 patients and hypothyroid in 7 and 2 patients, while 9 and 2 remain hyperthyroid, respectively.Serum free triiodothyronine, free thyroxine, and thyroid-stimulating hormone were statistically significant(P<0.05) before and after iodine-131 therapy, while no significant difference for serum thyrotrophin receptor antibody, antithyroid peroxidase autoantibody, and anti-thyroglobulin antibody.Atrial fibrillation was the most common cardiac complication of hyperthyroidism(n=25, 62.5%) .The remission rate after iodine-131 treatment was 76.0%.

CONCLUSIONIodine-131 therapy can effectively and timely control hyperthyroid in HHD patients.

Adult ; Heart Diseases ; drug therapy ; etiology ; Humans ; Hyperthyroidism ; complications ; drug therapy ; Iodine Radioisotopes ; therapeutic use ; Middle Aged

Objective To observe the effects of Yukouning on immunologic function in recurrent aphthous ulcer (RAU) patients with brimming heat of heart and spleen (BHHS type).Methods The study was composed of 30 patients with RAU (BHHS type) and 30 healthy individuals.All patients with RAU (BHHS type) were divided into levomisole group and Yukouning group (n =15).Patients of levomisole group were give 50 mg of levomisole,bid,lasted two days.Patients of Yukouning group were give Yukouning decoction.The changes of the class of T-cells and the levels of immunoglobulins (IgA,IgG,IgM) and complement (C3,C4) in different groups were tested by flow cytometry (FCM) and single immunodiffusion method.Results The levels of CD8+,IgG,IgA,C3 and C4 in RAU (BHHS type) patients increased significantly as compared with those in healthy group (P ＜ 0.05).The level of CD4+ and the ratio of CD4+/CD8+ decreased significantly as compared with those in healthy group (P ＜0.05).Furthermore,the levels of CD8+,IgG,IgA,C3 and C4 in RAU (BHHS type) patients decreased significantly after different drug treatment,respectively (P ＜ 0.05).At the same time,the level of CD8+ and the ratio of CD4+ / CD8+ increased significantly after different drug treatment,respectively (P ＜ 0.05).But all the above-mentioned indexes had no significant differences between the two drug groups (P ＞ 0.05).After 1-year follow-up,the effective rate (both 93.3％) had no significant difference between the two groups (P ＞ 0.05).Conclusion Imbalance of immune function is correlated with RAU (BHHS type).Yukouning plays a therapeutic role by regulating cellular and humoral immunity in RAU (BHHS type) patients.

This study is to evaluate the effects of the metformin (Met) on β cell function of diabetic KKAy mice. Female diabetic KKAy mice selected by insulin tolerance test (ITT) were divided randomly into two groups. Con group was orally administered by gavage with water, Met group with metformin hydrochloride at a dose of 0.2 g x kg(-1) for about 12 weeks. ITT and glucose tolerance tests (OGTT) were determined. Beta cell function was assessed by hyperglycemic clamp. Pancreatic biochemical indicators were tested. The changes of gene and protein expression in the pancreas and islets were also analyzed by Real-Time-PCR and immunostaining. Met significantly improved glucose intolerance and insulin resistance in KKAy mice. Fasting plasma glucose and insulin levels were also decreased. In addition, Met markedly increased glucose infusion rate (GIR) and elevated the Ist phase and maximum insulin secretion during clamp. It showed that Met decreased TG content and iNOS activities and increased Ca(2+) -Mg(2+)-ATPase activity in pancreas. Islets periphery was improved, and down-regulation of glucagon and up-regulated insulin protein expressions were found after Met treatment. Pancreatic mRNA expressions of inflammation factors including TLR4, NF-κB, JNK, IL-6 and TNF-α were down-regulated, p-NF-κB p65 protein levels also down-regulated by Met. And mRNA expressions of ion homeostasis involved in insulin secretion including SERCA2 and Kir6.2 were up-regulated by Met. Met increased SIRT5 expression level in pancreas of KKAy mice under the hyperglycemic clamp. These results indicated that chronic administration of Met regulated pancreatic inflammation generation, ion and hormone homeostasis and improved β cell function of diabetic KKAy mice.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; drug therapy ; Down-Regulation ; Female ; Glucose Tolerance Test ; Homeostasis ; Inflammation ; drug therapy ; Insulin ; secretion ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; Interleukin-6 ; metabolism ; Metformin ; pharmacology ; Mice ; NF-kappa B ; metabolism ; Pancreas ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism

[Summary] The relationship between the state of cognition and hypertension in 155 type 2 diabetic patients was studied.The results showed that significant difference in the state of cognition was found in patients with or without hypertension,so as to those with poorly or well controlled hypertension (P＜0.05).Systolic blood pressure,pulse pressure,mean arterial blood pressure,and duration of hypertension were negatively correlated with multiple cognitive domains,suggesting that proper control of blood pressure may have a protective effect on cognitive function in type 2 diabetic patients.

OBJECTIVETo study the effect of Mudan Granule (MD) on the glucose metabolism and beta cell function in monosodium glutamate (MSG) induced obese mice with insulin resistance (IR).

METHODSMSG obese mice were induced by subcutaneous injecting MSG (4 g/kg for 7 successive days in neonatal ICR mice). Forty MSG mice with IR features were recruited and divided into four groups according to body weight, fasting blood glucose, triglyceride (TG), total cholesterol (TC), and the percentage of blood glucose decreased within 40 min in the IR test, i.e., the model group (Con), the low dose MD group, the high dose MD group, and the Metformin group (Met). Besides, another 10 ICR mice were recruited as the normal control group (Nor). The water solvent of 2.5 g/kg MD or 5 g/kg MD was respectively administered to mice in the low dose MD group and the high dose MD group. Metformin hydrochloride was given to mice in the Met group at 0.2 g/kg body weight. Equal dose solvent distilled water was administered to mice in the Nor group and the Con group by gastrogavage, once per day. All medication was lasted for 15 weeks. Insulin tolerance test (ITT) and oral glucose tolerance test (OGTT) were performed after 6 weeks of treatment. Beta cell function was assessed by hyperglycemic clamp technique. The morphological changes in the pancreas were evaluated by hematoxylin-eosin (HE) staining. Changes of iNOS, NF-kappaB p65, and p-NF-kappaB p65 in the pancreas were tested.

RESULTSCompared with the Nor group, the blood glucose level, AUC, and fasting blood insulin, ONOO-contents, iNOS activities, and the expression of iNOS, NF-kappaB p65 subunit, pNF-kappaB p65 subunit obviously increased; decreased percentage of blood glucose within 40 min in ITT, glucose infusion rate (GIR), Clamp 1 min insulin, and Max-Insulin obviously decreased in the Con group (P < 0.05, P < 0.01). Compared with the Con group, the aforesaid indices could be improved in the Met group (P < 0.05, P < 0.01). In the low dose MD group, AUC, iNOS activities, and the expression of iNOS and p-NF-kappaB p65 subunit obviously decreased; percentage of blood glucose within 40 min in ITT and GIR obviously increased (P < 0.05, P < 0.01). In the high dose MD group, AUC, ONOO-contents, iNOS activities, and the expression of iNOS, NF-kappaB p65 subunit, and p-NF-KB p65 subunit obviously decreased; percentage of blood glucose within 40 min in ITT, Max-Insulin, and GIR obviously increased (P < 0.05, P < 0.01).

CONCLUSIONMD could significantly improve IR and functional disorder of 3 cells in MSG obese mice, which might be associated with lowering inflammatory reaction in the pancreas.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Insulin Resistance ; Insulin-Secreting Cells ; drug effects ; metabolism ; Male ; Metformin ; pharmacology ; Mice ; Mice, Inbred ICR ; Mice, Obese ; Obesity ; chemically induced ; metabolism ; Pancreas ; cytology ; drug effects ; Sodium Glutamate

OBJECTIVETo observe the effects of kaempferol and quercetin on the activity of cytochrome P450 in rat hepatocytes.

METHODSPrimarily cultured rat hepatocytes were exposed to kaempferol or quercetin in concentrations of 0.1, 1, 10 micromol/L for 12 h, 24 h and 48 h. Hepatocytes CYP isoemzymes-erythromycin N-demethylase (ERND) and aminopyrine N-demethylase (ADM) activities were determined by Nash methods. Erythromycin (10 micromol/L) was used as positive control and DMSO(0.1%) as solvent control.

RESULTSKaempferol and quercetin inhibited ENRD activity in a dose-and time-dependent manner. In dose-response study, the ENRD activities in kaempferol (0.1,1 and 10 micromol/L) treated groups were (0.088+/-0.008), (0.074+/-0.006) and (0.041+/-0.003)micromol/(mg.min(-1)), respectively. ENRD activity in quercetin treated groups at the same concentrations were (0.082+/-0.007), (0.063+/-0.007) and (0.034+/-0.005) micromol/(mg.min(-1)), respectively. In time-courses study, the ENRD activity exposed to 10 micromol/L kaempferol or quercetin for 12 h and 48 h were (0.053+/-0.006) and (0.037+/-0.007) micromol/(mg.min(-1)), or (0.067+/-0.005) and (0.032+/-0.004) micromol/(mg.min(-1)). ADM activity was inhibited only by kaempferol in 10 mol/L at 24 h, but was not significantly altered by quercetin at any concentration tested.

CONCLUSIONIn the present condition, kaempferol and quercetin act as potential CYP3A4 inhibitors as they can significantly inhibit ENRD in primarily cultured rat hepatocytes.

Aminopyrine N-Demethylase ; metabolism ; Animals ; Carcinoma, Hepatocellular ; enzymology ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Dose-Response Relationship, Drug ; Hepatocytes ; metabolism ; Kaempferols ; pharmacology ; Liver Neoplasms ; enzymology ; pathology ; Quercetin ; pharmacology ; Rats ; Tumor Cells, Cultured

OBJECTIVETo explore relationship between HBV DNA level and peripheral blood follicular helper T lymphocyte (Tfh) in patients with chronic hepatitis B (CHB) and its significance.

METHODSHBV DNA levels of 179 cases of CHB patients with positive HBV DNA, positive HBeAg and positive human leukocyte antigen(HLA)-A2 were tested with real time fluorescent quantitative PCR. Tfh and HBV specific CTL were tested with flow cytometry. IL-21 was also tested. 179 cases of CHB patients were divided into group A and group B based on HBV DNA levels, 86 cases in group A, HBV DNA levels were 10(4)-10(5) copies/ml, 93 cases in group B, HBV DNA levels were 10(6)-10(7) copies/ml. Above testing indexes of the two groups were compared.

RESULTSHBV DNA levels of group A were (4.85 +/- 0.37) log10 copies/ml, HBV DNA levels of group B were (6.83 +/- 0.31 ) log10 copies/ml, t = 27.31, P < 0. 001; Tfh of group A was (5.96 +/- 1.59)%, higher than that of group B (3.71 +/- 2.15)%, t = 4.92, P < 0.01; IL-21 of group A was (42.61 +/- 15.11)ng/L, higher than that of group B (14.91 +/- 3.15) ng/L, t = 8.62, P < 0.01; HBV specific CTL of group A was (0.36 +/- 0.08)%, higher than that of group B (0.18 +/- 0.06)%, t = 19.99, P < 0.001.

CONCLUSIONSerum HBV DNA level of CHB patients is related to the level of peripheral blood Tfh level: patients with low HBV DNA level have high Tfh level, high IL-21 level and high HBV specific CTL level. Patients with high HBV DNA level have low Tfh level, low IL-21 level and low HBV specific CTL level. The mechanism of baseline HBV DNA level affecting anti-viral therapy may be related to Tfh level.

Adult ; CD4 Lymphocyte Count ; DNA, Viral ; blood ; genetics ; Female ; HLA-A2 Antigen ; immunology ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; blood ; immunology ; virology ; Humans ; Interleukins ; immunology ; Male ; T-Lymphocytes, Helper-Inducer ; cytology