Animals ; Epitopes, T-Lymphocyte ; immunology ; Equine Infectious Anemia ; immunology ; virology ; Horses ; Infectious Anemia Virus, Equine ; immunology ; physiology ; T-Lymphocytes, Cytotoxic ; immunology ; physiology

OBJECTIVETo investigate the effect of Buyang Huanwu Decoction (BYHWD) and the different combinations of its ingredients on neurogenesis following ischemic stroke in rats.

METHODSThe model rats of ischemic stroke was established by blocking cerebral media artery with electrocoagulation through craniectomy, and electric stimulation, given from 24 h after blocking, 2 h daily for 15 successive days. They were divided into four groups, Group A treated with saline, Group B treated with BYHWD, Group C treated with BYHWD but earthworm subtracted, and Group D treated with Danggui Buxue Decoction (DGBXD). The expression of 5-bromodeoxyuridine (BrdU) in cerebral tissue was determined by immunohistochemical method.

RESULTSLarge amount of BrdU immunoreactive cells presented in the hippocampal region of rats in Group B and C, densely arranged, partial in cluster, with the figure significantly different to that in Group A (P < 0.01), and the amount in the ischemic side was significantly more than that in the opposite side (P < 0.05). While comparing between Group A and D, the amount of BrdU immunoreactive cells in the hippocampal region showed insignificant difference (P > 0.05).

CONCLUSIONBYHWD has a effect in promoting neurogenesis better than DGBXD.

Animals ; Bromodeoxyuridine ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hippocampus ; drug effects ; metabolism ; pathology ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; drug therapy ; Male ; Neurons ; drug effects ; metabolism ; pathology ; Neuroprotective Agents ; pharmacology ; therapeutic use ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVESTo investigate the effects of FAK-related non-kinase (FRNK) on the apoptosis of hepatic stellate cells (HSC) in vitro and on the extracellular signal-regulated kinase (ERK) signal transduction pathway.

METHODSHSC were stimulated by fibronectin (FN), and then they were transfected with FAK-related non-kinase (FRNK) plasmids mediated by cationic liposome. The apoptosis of FRNK-induced HSC was examined by annexin-V/propidium iodide double-labeled flow cytometry (FCM), gel electrophoresis and transmission electron microscopy. Levels of FRNK, FAK, p-FAK (Tyr397), ERK1 and p-ERK in HSC were assayed by Western blot on the protein level, and by RT-PCR on the mRNA level.

RESULTSThe expression of FRNK was enhanced after FRNK plasmids were transiently transfected into the HSC. The apoptotic rate of the HSC exposed to FRNK plasmids for 48 h was higher than that in the non-FRNK plasmid group (25.37%+/-1.92% vs 9.28%+/-1.05%, P less than 0.01), and was accompanied by a significantly higher activity of caspase-3 both in the protein and in the mRNA levels [(264.17+/-12.60 vs 185.82+/-9.69), P less than 0.01; (4.19+/-0.48 vs 1.07+/-0.27), P less than 0.01]. After exposure of HSC to FRNK plasmids, compared with the non-FRNK plasmid group, the expressions of p-FAK, ERK1 and p-ERK in protein and mRNA levels were lower; on the contrary, compared with the control group, the expressions of p-FAK, ERK1 and p-ERK in the FN group were higher.

CONCLUSIONThe expression of FRNK was enhanced and the phosphorylation of FAK was inhibited after FRNK was transiently transfected into HSC in vitro. FRNK induces apoptosis of HSC. FAK-ERK signal transduction pathway perhaps is involved in the process.

Apoptosis ; Cell Line ; Cell Proliferation ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hepatic Stellate Cells ; metabolism ; pathology ; Humans ; Protein-Tyrosine Kinases ; genetics ; metabolism ; RNA, Messenger ; genetics ; Signal Transduction

OBJECTIVETo determine the effects of Jiji decoction (Traditional Chinese Medicine) on the cognitive function and oxidative stress in mice with vascular dementia (VD) induced by cerebral ischemia/reperfusion.

METHODSThirty-two mice were randomly divided into nonnal group (n = 8), sham group (operation, but no cerebral ischemia/reperfusi6n, n = 8), model group (vascular dementia model induced by cerebral ischemia/reperfusion, n = 8), and Jiji decoction-treated group (vascular dementia model plus treatment with Jiji decoction, n = 8). Fourteen days of treatment after operation, the cognitive behavior was measured in step-through test, spatial probe test and platform test. Afterwards, to assess the levels of oxidative stress, the activity of superoxide dismutase(SOD) and content of malonaldehyde (MDA) in brain of these mice were measured.

RESULTSData from step-through test indicated that the escaping latency of Jiji decoction-treated group was prolonged and the error counts were decreased significantly ( P <0.01) compared with those of model group. Data from spatial probe test indicated that the time of entering darkroom, the time of climbing height and the time of entering bright room in Jiji decoction-treated group were shortened and the counts of climbing height were increased (P < 0.05-0.01) significantly compared with those of model group. Data from platform test showed that the escaping latency of Jiji decoction-treated group was prolonged significantly (P < 0.01) compared with that of model group. Compared with normal and sham group, the activity of SOD was decreased and the content of MDA was increased in model group significantly (P < 0.01). Compared with those of model group, the levels of SOD and MDA in Jiji decoction-treated group were improved significantly (P < 0.01).

CONCLUSIONJiji decoction could improve cognitive function of VD mice. Its mechanism might be related with the inhibition of oxidative stiess in the brain.

Animals ; Brain ; drug effects ; metabolism ; Cerebral Infarction ; physiopathology ; Cognition ; drug effects ; Dementia, Vascular ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Malondialdehyde ; metabolism ; Medicine, Chinese Traditional ; Mice ; Oxidative Stress ; drug effects ; Reperfusion Injury ; drug therapy ; Superoxide Dismutase ; metabolism

To analyze the characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy (DN) and its compliance with traditional Chinese medicine (TCM)symptom, for the sake of providing a basis for clarifying the rules of TCM syndrome differentiation in DN. Adopting the traditional epidemiological retrospective method, thirty-eight TCM syndromes and urinary protein with medium or low molecular weight, as well as urinary enzyme, including 24 h urinary protein (Upro), urinary albumin( UAlb), urinary retinal binding protein( URBP), urinary cystatin C (UCysC), urinary N-acetyl-beta-D-glucosaminidase (UNAG), were collected from 108 patients with stage III DN, and a multiple factor regression analysis between them was conducted. As the results, the levels of Upro, UAlb, URBP, UCysC, and UNAG were increased in 108 patients with stage III DN. Qi-Yin deficiency type was the major type. The level of UAlb in patients with Qi-Yin deficiency type was significantly higher than those without Qi-Yin deficiency type (P < 0.05). The elevation of Upro with the factors as swift digestion with rapid hungering, lassitude and lack of strength, weakness of waist and knees was complied, the elevation of UA1b with the factors as dry mouth with desire to drink, the elevation of URBP with the factors as numbness of extremities, shortness of breath, the elevation of UCysC with the factors as clear urine in large amounts, and the elevation of UNAG with the factors as frequent micturition, were complied respectively. In conclusion, for 108 stage III DN patients. The increase in urinary protein spectrum including UAlb, URBP, UCysC, and UNAG is the major characteristic. Shen and Pi are the major organs related to the appearance of urinary protein; Pi-Shen deficiency is the basic pathogenesis. The level of UAlb is taken as one of the objective syndrome factors for Qi-Yin deficiency type. The levels of UNAG and UCysC are possibly the objective syndrome factors for Shen-Qi deficiency type.
Diabetic Nephropathies ; complications ; diagnosis ; urine ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Proteinuria ; complications ; urine ; Qi ; Regression Analysis ; Yin-Yang

In clinic, some urinary protein makers can dynamically and noninvasively reflect the degree of renal tubular injury in patients with diabetic nephropathy (DN). These urinary biomarkers of tubular damage are broadly divided into two categories. One is newfound, including kidney injury molecule-1 (Kim-1), neutrophil getatinase-associated lipocalin (NGAL), liver-type fatty acid-binding protein (L-FABP) and cystatin C (CysC); the other one is classical, including beta2 microglobulin (beta2-MG), retinal binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (NAG). It is reported that, the increases in urinary protein markers are not only closely related to the damage of tubular epithelial cells in DN patients, but also can be ameliorated by the treatment with Chinese herbal compound preparations or Chinese herbal medicine. Recently, although urinary proteomics are used in the protein separation and identification, the traditional associated detection of urinary protein markers is more practical in clinic. At present, it is possible that the associated detection of urinary biomarkers of glomerular and tubular damages may be a feasible measure to reveal the clinical significance of urinary protein markers in DN patients and the interventional effects of Chinese herbal medicine.
Biomarkers ; urine ; Diabetic Nephropathies ; complications ; drug therapy ; urine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; Proteinuria ; complications

OBJECTIVETo observe the preventive effect of rats' serum containing Buyang Huanwu Decoction (BYHWD) on cultured cardiomyocyte apoptosis of neonatal rat induced by means of 24 hrs hypoxia and 4 hrs reoxygenation, and to investigate its mechanism concerned with nitric oxide (NO).

METHODSMyocyte apoptosis was detected by flow cytometry and ELISA with Annexin V-PI double labeled method. The lactate dehydrogenase (LDH) releasing level was measured with ultraviolet spectrophotometer. The NO concentration was determined by modified Yu method and the concentration of thiobarbituric acid response substance (TBARS) was tested by Ohkawa method.

RESULTSBYHWD contained rats' serum could significantly prevent cardiomyocyte from apoptosis induced by hypoxia and reoxygenation. After hypoxia-reoxygenation, the NO, LDH and TBARS levels in the supernatant of cultured liquid treated with BYHWD were significantly lower than those in non-treated cultured liquid, the effect of BYHWD was dose-dependent.

CONCLUSIONBYHWD can prevent cardiomyocytes from apoptosis induced by hypoxia and reoxygenation, its mechanism might be related with oxygen free radical and NO scavenging produced during the hypoxia-reoxygenation process.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Myocytes, Cardiac ; cytology ; drug effects ; Nitric Oxide ; metabolism ; Oxygen ; pharmacology ; Phytotherapy ; Rats ; Rats, Wistar

Glomerular hypertrophy is the main pathological characteristic in the early stage of diabetic nephropathy (DN), and its regulatory mechanism is closely related to mammalian target of rapamycin (mTOR) signaling pathway activity. mTOR includes mTOR complex 1 (mTORC1) and mTOR complex 2(mTORC2), in which, the upstream pathway of mTORC1 is phosphatidylinositol-3-kinase (PI3K)/serine-threonine kinase(Akt)/adenosine monophosphate activated protein kinase(AMPK), and the representative signaling molecules in the downstream pathway of mTORC1 are 4E-binding proteins(4EBP) and phosphoprotein 70 S6Kinase(p70S6K). Some Chinese herbal extracts could improve cell proliferation via intervening the expressions of the key molecules in the upstream or downstream of PIK/Akt/mTOR signaling pathway in vivo. As for glomerular mesangial cells(MC) and podocyte, mTOR plays an important role in regulating glomerular inherent cells, including adjusting cell cycle, energy metabolism and matrix protein synthesis. Rapamycin, the inhibitor of mTOR, could suppress glomerular inherent cell hypertrophy, cell proliferation, glomerular basement membrane (GBM) thickening and mesangial matrix deposition in model rats with DN. Some Chinese herbal extracts could alleviate glomerular lesions by intervening mTOR signaling pathway activity in renal tissue of DN animal models or in renal inherent cells in vivo and in vitro.
Animals ; Diabetic Nephropathies ; drug therapy ; enzymology ; genetics ; pathology ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertrophy ; drug therapy ; enzymology ; genetics ; pathology ; Kidney Glomerulus ; drug effects ; metabolism ; pathology ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

OBJECTIVETo explore the effect of Buyang Huanwu decoction (BHD) drug serum on rat's in vitro cultured cerebral cortical neuron apoptosis induced by hypoxia, and on the expression of p53 and p21 genes in hypoxia process.

METHODSThe model of hypoxia neuron apoptosis was established adopting Daniel method and treated with BHD drug serum. The neuron apoptosis rate was determined by flow cytometry with propidium iodide staining, the p53 and p21 gene expression was tested by immunohistochemical method with flow cytometry.

RESULTSBHD could significantly inhibit the neuron apoptosis induced by hypoxia and down-regulate the expressions of p53 and p21 genes.

CONCLUSIONBHD shows inhibition on neuron hypoxia apoptosis and down-regulating of the p53 and p21 gene expression is one of its mechanisms.

Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Male ; Neurons ; pathology ; Oncogene Protein p21(ras) ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Serum ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

Background and Objective:It has been proven that Ezrin protein may interact with E-cadherin protein and take part in metastasis of tumor cells.This study was to investigate the expressions of Ezrin and E-cadherin in esophageal squamous cell carcinoma(ESCC)and their relationship with the clinicopathologic factors,and to analyze their diagnostic values for ESCC.Method:The expression of Ezrin and E-cadherin in 72 specimen of ESCC and the para-cancer normal squamous epithelium was detected using tissue array with SP immunohistochemistry.Their correlations with the clinicopathologic factors were analyzed statistically.Results:The positive rate of Ezrin was significantly higher in ESCC than in para-cancer normal squamous epithelium (90.7%vs.46.0%,P＜0.001);the positive rate of E-cadherin was significantly lower in ESCC than in para-cancer normal squamous epithelium (27.6%vs.97.4%.P＜0.001).Ezrin expression was related to the invasiveness and lymph node metastasis of ESCC(P＜0.05);E-cadherin expression was related to the differentiation and lymph node metastasis of ESCC(P＜0.05).The high expression of Ezrin was related to the low expression of E-cadherin(P＜0.05).Conclusion:The activation of Ezrin and the absence of E-cadherin contributes to the tumorigenesis and metastasis of ESCC.