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

Animals ; Arginine Vasopressin ; metabolism ; Female ; Hippocampus ; drug effects ; metabolism ; Lead ; toxicity ; Learning ; drug effects ; Memory ; drug effects ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the utility of flow cytometry (FCM) in diagnosis and subclassification of non-Hodgkin lymphoma (NHL). Methods The samples of lymph nodes biopsy from 59 cases clinically suspected of NHL were detected by flow cytometry; and clonal lymphocytes and their immunophenotypes were identified analyzed. The concordance between the results of flow cytometry and histopathology was analyzed. Results Among the 59 cases, flow cytometry was able to identify aberrant clonal lymphocytes in 24 of 28 NHL cases identified by histopathology, the neoplastic lymphocytes ranged from 4.28 % to 89.10 %; 23 cases were diagnosed as B-NHL and 1 case was diagnosed as T-NHL. Compared with histopathology, the accuracy of FCM was 85.71% in diagnosis of NHL. The specificity and sensitivity of FCM was 100 % and 92% in diagnosis of B-NHL. The accuracy of flow cytometry immunophenotyping in classification of 24 cases of NHL was consistent with that of histopathology. Conclusion Flow cytometry could be an ancillary technique in diagnosis of NHL by identifying aberrant clonal lymphocytes, and enable identification of B-NHL subtype.

Objective To investigate the clinical value of multi-slice spiral CT (MSCT)in the diagnosis of focal pulmonary ground-glass opacity nodules(fGGO)of 1 cm or less.Methods The MSCT examination data of 95 patients with subcentimeter fGGO was analyzed and compared with pathology results.The different pathological types of fGGP lesion size,internal solid component size,mixed type ground glass nodules (mGGO)proportion and pleural sag,lesion shape,grave leaves,burr,cavitation and the boundary conditions were compared. According to the relationship of the lesions and the surrounding blood vessels,3 types were classified,and the relationship of fGGO and blood vessel were analyzed.Results The minimal lesion was 0.41 cm of 95 cases and the maximum was 0.99 cm.There was statistically signifi-cant difference in the lesion size,solid component sizes,mGGO and lobulation occupied the percentage of the group before infiltrating and the adenocarcinoma group (P <0.05),and there was no statistically significant difference in pleural indentation,lesion shape,burr,vacuoles and boundary conditions (P >0.05).The difference was statistically significant in the leaflet occupied the percentage of the benign group com-pared with the adenocarcinoma group,and the differences of the remaining features were not statistically significant compared with the adeno-carcinoma group(P >0.05).According to the relstionship between fGGO and the vessels,patients in the benign group were type Ⅰ or typeⅡ,type Ⅲ was found in 10 cases of the group before infiltrating (25.6%),and type Ⅲ in 17 cases of the adenocarcinoma group (39.5%), the proportion of type Ⅲ of the group before infiltrating and the adenocarcinoma group was higher than that of the benign group (P <0.05). Conclusion The qualitative diagnosis of pulmonary fGGO is difficult,and the relationship between the morphology of the lesions under the MSCT and the surrounding vessels have some value for the diagnosis of the lesions.

Objective To investigate the clinical pathologic significance of lymphangiogenesis in in colorectal cancer. Methods New lymphatic-specific markers D2-40 was used immunohistochemically to detect the lymphatic vessel density(LVD) in the intratumoural and peritumoral areas, and in normal tissue from 96 cases of colorectal cancer, which were analyzed with clinical pathologic parameters of those colorectal cancer. Results Significandy higher LVD was found in the intratumoural area(14.5±2.4), when compared with normal(5.9±1.1)and peritumoural areas(6.7±1.2) (P<0.01). LVD of the peritumoural area was higher than normal area (P< 0.01). However, peritumoural LVD was associated with both depth of invasion and liver metastasis (r=0.71,0.78 P<0.05), but not associated with tumour size, macroscopic type and lymph-node metastasis (P>0.05). Intratu-moural LVD was not correlated with tumour size, macroscopic type, the depth of invasion,lymph-node metastasis, and liver metastasis(P>0.05). Conclusion Lymphangiogenesis in the peritumoural area may be helpful in evalution of liver metastasis and prognosis.

Objective To study the expressions of YKL-40 and IL-1β in the cartilage of knee osteoarthritis(KOA),and analysis the possible relationship between YKL-40,IL-1β in KOA.Methods Thirty-eight patients diagnosed with KOA were selected in this hospital as observation group.Then select 30 articular cartilage cases in the same period due to knee injury knee joint examina tion or treatment of knee joint fracture surgery patients,referred to as the control group.According to KOA radiation and arthroscopic grading standards,the observation group was divided into 16 cases of mild group,10 cases of moderate group,11 cases of severe groups.Compare the expression levels of all patients including YKL-40,interleukin-1β (IL-1β),IL 6,tumor necrosis factor-α (TNF-α),while recording Mankin score and cell mortality.Results Expression of YKL 40,IL-1β,IL-6,TNFα,Mankin scores and rate of cell death about observation group were significantly higher than control group,the indicators above in mild,moderate,severe group showed a trend of rising(all P＜0.05).And the expression levels of YKL-40 and IL-1β were significantly positive correlation (r=0.738,P=0.000).In addition, The expression levels of YKL-40 had relation with IL 6 (r=0.819,P=0.000),TNF-α (r=0.871,P=0.000) and Mankin score (r=0.832,P 0.000),cells mortality deposit (r=0.832,P=0.000).Conclusion Expression levels of YKL-40 and IL-1β were significantly increased in cartilage of knee osteoarthritis patients,and there showed a significant positive correlation between YKL-40 and IL-1β.

Objective To study the effect of the extract of Chrysanthemum Indicum(CI)on the expression of tumor necro-sis factor-α (TNF-α) and neutrophil function in chronic bronchitis (CB)rats and to explore the therapeutic mechanism for chronic bronchitis. Methods The extract of CI was prepared. Wistar rats were randomly divided into blank control group, model group, Chuanxiling group, and low-, middle-and high-dose CI extract groups. Rat model of CB was established by intratracheal injection with low-dose lipopolysaccharide. After drug intervention, the expression of TNF-α in rat serum and bronchoalveolar lavage fluid was detected by ELISA method. Phagocytic function of the neu-trophil and respiratory burst of the rats were measured by flow cytometry (FCM). Results Compared with the blank con-trol group, the phagocytic function of the neutrophil, respiratory burst of the rats, and the expression of TNF-α in rat serum and bronchoalveolar lavage fluid of the model group were increased significantly. CI extract significantly decreased the abnormal rising of the above indexes. Conclusion Down-Regulation of TNF-α expression, and decrease of the neutrophil phagocytic function and rat respiratory burst may be one of the therapeutic mechanisms of Chrysanthemum In-dicum extract for the treatment of CB.

【Objective】To study the effect of the specific p38 mitogen-activated protein kinase(p38 MAPK) inhibitor SB203580 on apoptosis of cerebellar granule neurons induced by low potassium.【Methods】Apoptosis was induced by switching the cultured cerebellar granule neurons from a culture medium containing K+ 25 mmol*L-1 to a medium containing K+ 5 mmol*L-1 (cLK).Fragmentation of DNA was analyzed using agarose gel eletrophoresis.SAPK/JNK activity was measured by SAPK/JNK assay kit.【Results】Low potassium resulted in apoptosis as characterized by morphological and biochemical features,but the specific p38 MAPK inhibitor SB203580 improved the survival of cerebellar granule neurons cultured in cLK medium by blocking apoptosis in a concentration-dependent manner.The expression and phosphorylation of c-Jun increased and the activity of c-Jun N-terminal protein kinase (JNK) elevated when cerebellar granule neurons were cultured in cLK medium.But when the cerebellar granule neurons cultured in cLK medium were exposed to 25 μmol*L-1 SB203580,the expression and phosphorylation of c-Jun and the activity of JNK were both decreased evidently.【Conclusions】These results indicate that SB203580 inhibits the activation of JNK and phosphorylation of c-Jun,and therefore protects granule neurons from apoptosis induced by low potassium.

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

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