To study the compatible mechanisms and compatible proportion of Shaoyao Gancao decoction, the intestinal absorption of main ingredients in Shaoyao Gancao decoction SG11 (Baishao-Zhigancao 1: 1) , SG31 (Baishao-Zhigancao 3: 1), Baishao water decoction S and Zhigancao (G) were investigated and compared using in vitro everted intestinal sac model and in situ single pass intestinal perfusion (SPIP) model. The concentration of paeoniflorin (PF), liquiritin (LQ) and mono-ammonium glycyrrhizinate (GL) in test samples and samples of intestinal sac and intestinal perfusion was determined by HPLC. The intestinal absorptive amount and absorption parameters were calculated. Results showed that in the everted intestinal sac model, three ingredients could be absorbed by duodenum, jejunum and ileum, and the absorption in the jejunum was best for all 3 ingredients. The absorption rate of three ingredients in SG11 was significantly higher than that in single decoction (P < 0.05), but had no significant difference compared with SG31. In SPIP model, the absorption rate constant K(a), the apparent absorption coefficient P(app) and the absorption rate of three ingredients in SG11 were significantly higher than those in single decoction. Parameters of PF and GL in SG11 were significantly higher than those in SG31, but had no differences of LQ. It proved that the compatibility of Baishao and Zhigancao could improve the intestinal absorption of PF, LQ and GL. The absorption of each ingredient in SG11 was better than that in SG31.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; blood supply ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective To observe the change of capillary pericapillary cells in rats with myocardial infarction and the influence of supplementing qi and activating blood circulation herbs, and explore its mechanism of improving myocardial perfusion. Methods The rat model was established by ligaturing the left anterior descending coronary artery. On the base of ECG evaluation, successfully modeled rats were randomly divided into the model group, group treated with supplementing qi and activating blood circulation Chinese medicine (activating blood and supplementing qi group), group treated with Perindopril (Perindopril group), group treated with Tongxinluo Capsules (Tongxinluo group). The sham-operation group was taken as the control. There were totally 5 groups. The model group and the sham-operation group were treated with normal saline. The changes of myocardial capillary density (MCD) and number of pericapillary cells on the 7th, 28th day after medicinal administration were observed. Results On the 7th and 28th day, the MCD decreased significantly and the number of capillary pericapillary cells increased significantly in the model group compared with the sham-operation group (P<0.01). The MCD increased significantly in the activating blood and supplementing qi group, Perindopril group and Tongxinluo group compared with the model group (P<0.01). The number of pericapillary cells decreased significantly in the activating blood and supplementing qi group, Perindopril group and Tongxinluo group compared with the model group (P<0.01). Conclusion The supplementing qi and activating blood circulation herbs can improve regional myocardial blood supply by decreasing the number of pericytes and promoting regeneration of capillary.

Objective To determine the electrocardiographic (ECG) characteristics of myocardial infarction (MI) evolution in rats and the intervention effect of Chinese herbs, and to provide basis for the establishment of the criteria for ECG diagnosis and assessment of drug therapeutic effects of rats MI. Methods Totally 140 male SD rats were randomly divided into sham operation group, model group, replenishing qi group, activating blood group, replenishing qi and activating blood (1∶2) and (2∶1) group as well as Tongxinluo group, each group with 20 rats. The rat MI model was established by ligating the left anterior descending coronary artery. The treatment groups were administrered with corresponding drugs by gavage from the first day after operation. The sham operation group and model group were given the same volume of distilled water. The 12-lead ECGs were recorded before, immediately after, 1st and 2nd week after operation respectively. The voltage value of ST segment deviation, the time limit of QRS complex and the number of animals with pathologic Q wave were evaluated for statistical analysis. Results Model group showed the elevation of ST segment, significantly prolonged time limit of QRS complex (P<0.01), and no pathologic Q wave showed immediately after operation compared with sham operation group. And all above observational indexes reached the peak at 1st week and declined at 2nd week after operation. Compared with model group at 2nd week after operation, replenishing qi and activating blood (1∶2) group and (2∶1) group all presented remarkable dropping of ST segment, shortening in time limit of QRS complex and reduction in number of rats with pathologic Q wave, of which 2∶1 group showed the most (P<0.01). Activating blood group only displayed decreases in time limit of QRS and number of rats with pathologic Q wave (P<0.05), and no significant decline in ST segment. Replenishing qi group demonstrated no significant changes in above three observational indexes (P>0.05). Thus, we proposed the criterion for the ECG diagnosis of rats MI as well as the criterion for the ECG assessment of drug therapeutic effects of rats MI. Conclusion ECG can overall and sensitively evaluate the evolution and drug therapeutic effects of MI in rats, thus providing a relatively objective and available assessment method for the experimental studies of myocardial ischemic diseases.

OBJECTIVETo culture rat corpus cavernosum smooth muscle cells in vitro using a modified tissue culture method.

METHODSFifteen male rats were randomized into 3 equal groups, namely enzyme digestion group, tissue culture group, and modified tissue culture group. The penis of the rats was separated carefully and cut into small pieces, and seeded onto culture flasks and cultured in complete medium consisting of DMEM containing 20% fetal calf serum at 37 degrees C; in a humidified atmosphere with 5% carbon dioxide. The cells growth was observed under phase contrast microscope and the smooth muscle cell specific proteins alpha-SM-actin and desmin were identified immunohistochemically.

RESULTSThe alpha-SM-actin-positive cell rate was 96.3% in rat corpus cavernosum smooth muscle and 23.8% in the fibroblasts, and the corpus cavernosum smooth muscle contained 74.4% desmin-positive cells while the fibroblasts showed no desmin positivity. Significant difference was found in the positive cell rate for desmin among the 3 groups, with the highest positive cell rate occurred in modified tissue culture group.

CONCLUSIONDesmin may serve as a marker for identifying corpus cavernosum smooth muscle cells. The modified tissue culture method can result in highly purified corpus cavernosum smooth muscle cells with intact structure and functions.

Actins ; analysis ; Animals ; Biomarkers ; analysis ; Cell Proliferation ; Desmin ; analysis ; Male ; Muscle, Smooth ; cytology ; Penis ; cytology ; Rats ; Rats, Sprague-Dawley ; Tissue Culture Techniques

OBJECTIVETo investigate the expression of proline-rich tyrosine kinase-2 (Pyk2) in human primary colorectal carcinoma (CRC) and it's prognostic significance.

METHODSThe expression of Pyk2 was retrospectively examined with immunohistochemistry (IHC) in 108 tissues of primary CRC. The correlation of Pyk2 expression to prognosis and relevant clinical factors were analyzed.

RESULTSThe rate of Pyk2 low-expression in CRC was 56.5% (61/108). The expression of Pyk2 correlated significantly to the histological grade (P < 0.05) and the TNM stage (P < 0.05), while no correlation between Pyk2 expression and age, tumor size (P > 0.05). Patients with Pyk2 over-expression had significantly higher 5-year survival rate (66.0%) than those with Pyk2 low-expression (31.4%). Pyk2 expression, together with carcinoma histologic grade and TNM stage were prognostic factors to CRC on the multivariate analysis.

CONCLUSIONSPyk2 expression can be a prognostic factor to the CRC patients together with other predictors.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; enzymology ; pathology ; Female ; Focal Adhesion Kinase 2 ; metabolism ; Humans ; Male ; Middle Aged ; Prognosis

OBJECTIVETo explore the correlation between pathological characteristics of target organs and excess evil syndrome in IgA nephropathy.

METHODSData were collected in multicenter cooperation. Totally 266 IgA nephropathy patients were typed into exogenous wind-heat affection syndrome (49 cases), lower energizer damp-heat syndrome (100 cases), damp-phlegm syndrome (43 cases), and blood stasis syndrome (74 cases). Meanwhile, percutaneous renal biopsy was performed in all patients for Hass classification, Oxford classification, Katafuchi integral, and Jiang's classification methods. The correlation between excess evil syndrome and pathological index was analyzed.

RESULTSFour syndrome types were correlated with their Hass levels (r = 0. 341, P <0. 01). Affection of exogenous wind-heat syndrome was correlated with segmental proliferation of endothelial cells and damaged active lesions of segmental capillary loops. Lower-energizer damp-heat syndrome was associated with Hass III level, destroying active lesions of capillary loops, segmental proliferation of endothelial cells, glomerular segmental lesions, focal interstitial infiltration of inflammatory cells, focal interstitial fibrosis and tubular atrophy. Blood stasis syndrome was associated with Hass IV level, glomerular sclerosis, segmental glomerulosclerosis (S)/adhesion, mesangial hypercellularity (M), angiohyalinosis, multi-foci interstitial infiltration of inflammatory cells, multi-foci interstitial fibrosis and tubular atrophy. Phlegm-damp syndrome had higher proportions of Hass I and III levels, but with no association with other pathological parameters.

CONCLUSIONSExcess evil syndrome was associated with partial pathological characteristics of IgA nephropathy. It could reflect pathological damage degree of target organs, activities, chronic lesions, and prognosis of IgA nephropathy to certain extent. Correlated pathological characteristics and its evolution could indicate excess evil syndrome types and their evolution rules.

Capillaries ; Fibrosis ; Glomerulonephritis, IGA ; pathology ; Glomerulosclerosis, Focal Segmental ; Humans ; Kidney Glomerulus ; Medicine, Chinese Traditional ; Prognosis ; Syndrome

OBJECTIVETo investigate the method for culturing corpus cavernosum smooth muscle cells (CCSMs) derived from diabetic rats with erectile dysfunction (ED) for the study of ED caused by diabetes.

METHODSCCSMs were isolated from the corpus cavernosum of diabetic rats with ED and cultured using a modified method of adherent tissue culture. The cultured cells were identified by immunohistochemistry and the cell morphology and proliferation were observed.

RESULTSThe primary culture of CCSM was performed successfully, and the cells were seen to migrate from the small tissue pieces 3 days later, reaching nearly confluence in 16-18 days. A typical "hill-valley" growth pattern was noted in the cell passaging. Immunohistochemical staining for alpha-smooth muscle actin (alpha-SM-actin) and desmin yielded positive results in the cells.

CONCLUSIONThe modified method for adherent tissue culture is convenient and reliable in establishing the in vitro cell culture model of CCSMs from diabetic rats with ED, and the cultured CCSMs display a faster proliferation than normal CCSMs. No obvious differences in the cell morphology can be found between diabetic and normal CCSMs under light microscope.

Animals ; Cell Culture Techniques ; Cells, Cultured ; Diabetes Mellitus, Experimental ; complications ; pathology ; physiopathology ; Erectile Dysfunction ; etiology ; pathology ; physiopathology ; Male ; Models, Biological ; Myocytes, Smooth Muscle ; cytology ; physiology ; Penile Erection ; Penis ; cytology ; Rats ; Rats, Sprague-Dawley

Objective:To preliminarily verify the cross talk between tissue factor/active coagulation factor Ⅶ (TF/FⅦa) and epidermal growth factor receptor (EGFR) pathways in human colon cancer cells in culture.Methods:FⅦa was treated to HT-29 (KRAS-wild type) and LoVo (KRAS-mutant) colon cancer cells to activate TF/F Ⅶa pathway,qRT-PCR and Western blot were used to detect the expressions of amphiregulin (AREG) and epiregulin (EREG),ligands of EGFR on mRNA and protein levels,respectively.After knocking down expression of TF by TF-targeted siRNA transfection,FⅦa was treated and mRNA expressions of AREG and EREG were detected to see whether the FⅦa-induced effects were dependent on TF.Expressions of mRNA of TF and FⅦwere detected by qRT-PCR following the activation of EGFR pathway by treatment with epidermal growth factor (EGF) to HT-29 and LoVo cells.Results:After TF/FⅦa pathway was activated,for HT-29 cells,expressions of AREG (on mRNA level) and EREG (both on mRNA and protein level) were significantly down-regulated versus those of control group,gene expressions of AREG and EREG were 0.55 ± 0.09 vs.0.99 ± 0.09,0.67 ± 0.10 vs.1.02 ± 0.02,protein expressions of EREG were 0.54 ± 0.09 vs.1.04 ± 0.13,all P ＜ 0.05.For LoVo cells,expressions of AREG (both on mRNA and protein level) and EREG (on protein level) were significantly up-regulated versus those of control group,gene expression of AREG were 1.87 ± 0.39 vs.0.93 ± 0.23,protein expressions of AREG and EREG were 3.09 ±0.73 vs.1.11 ±0.21,1.53 ±0.19 vs.0.97 ± 0.23,all P ＜0.05.The regulating effect of AREG and EREG mRNA expression by FⅦa in HT-29 and LoVo cells could both be partly blocked by knocking down TF expression.For HT-29 cells,activation of EGFR pathway induced no significant TF mRNA expression,F Ⅶ mRNA expression was not detected.However,for LoVo cells,activation of EGFR pathway induced significantly higher mRNA expressions of both TF and FⅦ,expressions were 1.53 ± 0.23 vs.1.00 ± 0.23,53.20 ± 6.08 vs.1.00 ± 0.15,all P ＜0.05.Conclusion:In colon cancer cell LoVo,when activated,TF/FⅦa pathway and EGFR pathway could interact through upregulating the other pathway's effectors,and mutant KRAS might play a critical role in the two pathways'cross talk.

OBJECTIVETo investigate the clinical diagnosis, treatment and immunohistochemical characteristics of rectal stromal tumors.

METHODSImmunohistochemical expression of CD117 was retrieved in 20 cases of mesenchymal tumors of the rectum. And we analyzed the immunohistochemical characteristics, clinical data of rectal stromal tumors, and the results retrospectively.

RESULTSSixteen cases of rectal stromal tumors, 3 cases of leiomyosarcomas and 1 case of schwannoma were diagnosed. Histologically, 2 cases of GISTs were classified as benign, 1 as borderline and 13 as malignant. All of rectal stromal tumors (100%) were strongly positive for CD117, and 14 cases (88%) positive for CD34. The demographic profile of rectal stromal tumors showed a male predominance with average age of 60 years old. The main symptoms were urinary retention, constipation and abdominal pain. 14 cases were positive in digital rectal examination. The recurrence rate of local and radical resection in malignant stromal tumors was 4/4 and 3/6. 1, 3, 5-year survival rates were 89%, 64%, 48%, respectively. After operation mean survival time of was 47 months.

CONCLUSIONSThe specific GIST constituted the majority of mesenchymal tumors in rectum. It usually showed malignant biological behavior. Invasion and recurrence were common. Earlier diagnosis and radical resection had better prognosis. Periodically following up can help to detect the recurrence timely.

Adult ; Aged ; Female ; Gastrointestinal Stromal Tumors ; diagnosis ; pathology ; surgery ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Rectal Neoplasms ; diagnosis ; pathology ; surgery ; Retrospective Studies

BACKGROUNDStudies have shown that abnormal activation of the sonic hedgehog pathway is closely related to tumorigenesis in central nervous system. This study aimed to investigate the role of the sonic hedgehog signaling pathway in the occurrence of brainstem and supratentorial glioma.

METHODSReal-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry were used to detect the expression of sonic hedgehog-related components in 5 specimens of normal brain tissue, 10 of grade II brainstem glioma, and 10 of grade II supratentorial glioma. The significance of differences between two groups was determined using the Mann-Whitney U test or the two-sample test according to the results of normality distribution tests.

RESULTSThe mRNA expression levels of sonic hedgehog-related genes were higher in brainstem astrocytomas than in supratentorial astrocytomas and normal brain tissue. The level of protein patched homolog 1 (PTCH1) was significantly higher in brainstem astrocytomas than in supratentorial astrocytomas and normal brain tissue (P < 0.01). Immunohistochemistry semi-quantitative analysis was consistent with the qRT-PCR result that PTCH1 expression was increased significantly in brainstem astrocytomas at the protein level (P < 0.05).

CONCLUSIONSEnhanced PTCH1 expression and activation of the sonic hedgehog pathway are involved in brainstem glioma. This may be related to the difference in malignant biological behavior between brainstem and hemispheric glioma, and could be an ideal therapeutic target in brainstem glioma.

Adolescent ; Adult ; Astrocytoma ; genetics ; metabolism ; Brain Stem Neoplasms ; genetics ; metabolism ; Female ; Glioma ; genetics ; metabolism ; Hedgehog Proteins ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Patched Receptors ; Patched-1 Receptor ; Real-Time Polymerase Chain Reaction ; Receptors, Cell Surface ; genetics ; metabolism ; Signal Transduction ; genetics ; physiology ; Supratentorial Neoplasms ; genetics ; metabolism ; Young Adult