OBJECTIVETo observe the effect of Buyang Huanwu Decoction (BYHWD), a representative formula of qi benefiting blood activating method on aorta Rho associated coiled-coil forming protein serine/threonine kinase (Rhokinase, ROCK) and nuclear transcription factor kappa B (NF-κB) p65 mRNA expressions and levels of blood lipids in atherosclerosis (AS) model rats.

METHODSThe AS rat model was prepared by vitamin D3 and high fat diet. Totally 60 rats were randomly divided into 6 groups, i.e., the normal control group, the model group, the low dose BYHWD group (10 g/kg), the high dose BYHWD group (20 g/kg), the Simvastatin control group (0.6 mg/kg), and the BYHWD prevention group (10 g/kg), 10 in each group. After successful modeling all medication was intervened for 28 days. Expression levels oxidized low density lipoprotein (ox-LDL) were detected by ELISA. Levels of TG, TC, LDL-C, HDL-C were determined by enzyme method. Pathological changes of aortic tissue were observed under light microscope. mRNA expressions of Rho kinase and NF-κB p65 in aorta were detected by real time (RT) PCR.

RESULTSHigh fat diet and peritoneal injection of vitamin D3 could induce AS rat model. Typical atheromatous plaque formed in aorta of AS model rats. Compared with the normal control group, levels of TC, TG, LDL-C, and ox-LDL significantly increased in the model group, but the HDL-C level decreased (P < 0.01). Compared with the model group, levels of TC, TG, LDL-C, and ox-LDL all decreased, but HDL-C increased in low and high dose BYHWD groups, the Simvastatin control group, and the BYHWD prevention group (P < 0.05, P < 0.01). Compared with the low dose BYHWD group, above-mentioned indices were more obviously lowered in the high dose BYHWD group, the Simvastatin control group, and the BYHWD prevention group (P < 0.05). Compared with the normal control group, mRNA expression levels of Rho kinase and NF-κB p65 significantly increased in the model group (P < 0.01). Compared with the model group, mRNA expressions of Rho kinase and NF-κB p65 obviously decreased in low and high dose BYHWD groups, the Simvastatin control group, and the BYHWD prevention group (P < 0.01). Compared with the low dose BYHWD group, the two indicators were more obviously lowered in the high dose BYHWD group, the Simvastatin control group, and the BYHWD prevention group (P < 0.05). But there was no statistical difference in blood lipids levels, mRNA expression levels of Rho kinase or NF-κB p65 among the high dose BYHWD group, the Simvastatin control group, and the BYHWD prevention group (P >0. 05).

CONCLUSIONSBYHWD could down-regulate mRNA expression levels of Rho kinase and NF-κB p65, lower levels of blood lipids, and fight against AS. Suppressing Rho kinase pathway might be one of its mechanisms.

Animals ; Aorta ; Atherosclerosis ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Gene Expression ; drug effects ; Lipids ; Lipoproteins, LDL ; NF-kappa B ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Simvastatin ; Transcription Factor RelA ; metabolism ; rho-Associated Kinases ; metabolism

Objective To investigate the difference of hypoglycemic and hypolipidemic effects of Coptis deltoidea (Coptis chinensis) and its processed products on T2DM rats. Methods Type 2 diabetes mellitus induced by high sugar, high fat diet and low dose of streptozotocin. C. deltoidea were processed with yellow wine, ginger, and evodia rutaecarpa, respectively. The effects of C. deltoidea on fasting blood glucose (FBG), fasting insulin (FINS), glycosylated serum protein (GSP), pancreatic pathology, serum triglyceride (TG), total cholesterol (TC) and lipoprotein (HDL-C or LDL-C) and key protein and gene expression of SCAP\SREBP-1c pathway in liver of model rats were investigated. Result C. deltoidea and its processed products reduced the FBG and GSP in T2DM rats. The wine products significantly reduced FBG and GSP in model rats (P < 0.01). The hypoglycemic range was better than raw and ginger products. At hypolipidemic aspect, C. deltoidea and its processed products decreased the content of TG, TC, and LDL-C (P < 0.05, 0.01), increased the content of HDL-C in serum (P < 0.05, 0.01), and down-regulated the protein and gene expression of SCAP/SREBP-1c in liver, the insulin target organ. However, wine and evodia products were better than crude drug or ginger products. Conclusion C. deltoidea processed by yellow wine, ginger and evodia rutaecarpa is traditional method to improve the disadvantages of C. deltoidea with bitter cold nature and not easy to take for a long time. However, different processing methods have different effects. Clinical medication should be combied withTCM syndrome differentiation and simple hypoglycemic effect of wine product is appropriate; while wine and evodia products were better in hypoglycemic and lipid regulation aspects.

BACKGROUND: Representative formulae of Qi-Benefiting and Blood-Activating Chinese medicinal herbs Bu Yang Huan Wu Tang (BYHWT) can improve microcirculation and hypoxia, decrease fibrinogen, resist platelet aggregation, reduce blood lipid, dilate blood vessel, resist thrombosis, and reduce neonatal plaque rupture, and has a multi-target, multi-channel anti-atherosclerotic effect.
OBJECTIVE: To observe the influence of representative formulae of Qi-Benefiting and Blood-Activating Chinese medicinal herbs BYHWT on aorta Rho kinase, tissue factor, matrix metal oproteinase-2, matrix metal oproteinase-9 mRNA expression in atherosclerosis models, and to explore the action mechanism of BYHWT on anti-atherosclerosis.
METHODS: Vitamin D3 and high fat diet induced atherosclerosis in rats. 60 rats were randomly divided into normal control group, model group, 10 g/kg BYHWT therapy group, 20 g/kg BYHWT therapy group, simvastatin control group and BYHWT prevention group (n=10). At 28 days after model establishment, the expression level of Rho kinase, tissue factor, matrix metal oproteinase-2, matrix metal oproteinase-9 mRNA expression and blood lipid level were detected.
RESULTS AND CONCLUSION: (1) High fat diet and vitamin D3 could induce atherosclerosis in rats. Atherosclerotic plaque formed in the model group. (2) Rho kinase, tissue factor, matrix metal oproteinase-2, matrix metal oproteinase-9 mRNA expression and blood lipid were significantly lower in the normal control group, 20 g/kg BYHWT therapy group, simvastatin control group and BYHWT prevention group than in the model group (P < 0.01). Above indexes were significantly lower in the 10 g/kg BYHWT therapy group than in the model group (P < 0.05). No significant difference in above indexes was detected among BYHWT prevention group, 20 g/kg BYHWT therapy group and simvastatin control group (P > 0.05). (3) Results indicated that Qi-Benefiting and Blood-Activating BYHWT can resist atherosclerosis, down-regulate the expression of Rho kinase, tissue factor, matrix metal oproteinase-2, and matrix metal oproteinase-9 mRNA expression, which may be one of the mechanisms of anti-atherosclerosis.

OBJECTIVETo observe the pathological changes of renal tissue in the rats with paraquat (PQ) poisoning as well as the serum creatinine (SCr) levels and expression levels of hypoxia-inducible factor-1α (HIF-1α) and transforming growth factor-β (TGF-bgr;) in renal tissue at different time points after PQ poisoning, and to investigate the association of HIF-1α with renal injury after PQ poisoning.

METHODSForty-eight healthy male Sprague-Dawley rats were randomly divided into control group (n = 6) and PQ group (n = 42). The control group was given a single dose of 1 ml saline by gavage; the PQ group was given a single dose of 1 ml PQ (50 mg/kg), which was prepared by diluting 20% raw liquid of PQ with saline, by gavage. The PQ group was further divided into 2, 6, 12, 24, 48, 72 and 120 h PQ subgroups (n = 6 for each subgroup) to be examined at 2, 6, 12, 24, 48, 72, or 120 h after gavage. Their arterial blood was collected for blood gas analysis as well as blood lactic acid (BLA) and SCr measurement; renal sections were subjected to HE staining; the protein expression of HIF-1α and TGF-β in renal tissue was measured by Western blot.

RESULTSThe BLA level and SCr level began to rise at 6h after poisoning. Compared with the control group, the 6, 12, 24, 48, 72 and 120 h PQ subgroups had significantly increased BLA and SCr levels (P < 0.05); the 72 and 120 h PQ subgroup showed hypoxemia (P < 0.05). The protein expression of HIF-1α in PQ group increased significantly at 6h and reached the peak level at 72 h, with a significant difference from that in the control group at 6, 12, 24, 48, 72, and 120 h (P < 0.05). The protein expression of TGF-β in PQ group began to rise at 24 h, reached the peak level at 72 h, and declined at 120 h, with a significant difference from that in the control group at 24, 48, and 72 h (P < 0.05). The protein expression of HIF-1α was positively correlated with SCr level (r = 0.9308, P = 0.0008), uncorrelated with arterial partial pressure of oxygen (r = -0.6996, P = 0.0534), and positively correlated with BLA level (r = 0.9483, P = 0.0003). The pathological changes of renal tissue mainly included the degeneration and necrosis of renal tubular epithelial cells, which worsened as the time went on and appeared less severe at 120 h.

CONCLUSIONThe HIF-1α expression in renal tissue increases significantly in the early stage of PQ poisoning, which is associated with increased BLA and SCr levels and causes upregulated expression of TGF-β that promotes renal fibrosis.

Animals ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Kidney ; metabolism ; pathology ; Male ; Paraquat ; toxicity ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; metabolism

BACKGROUNDLeukocyte adhesion deficiency type 1 (LAD-1) is a rare, autosomal recessive inherited immunodeficiency disease characterized by recurrent severe bacterial infection, impaired pus formation, poor wound healing, associated with the mutation in the CD18 gene responsible for the ability of the leucocytes to migrate from the blood stream towards the site of inflammation. Correct and early diagnosis of LAD-1 is vital to the success of treatment and prevention of aggressive infections. The purpose of this study was to collect the clinical findings of the disease and to identify the genetic entity.

METHODSCD18 expression in the peripheral blood leukocytes from the patient, his parents and normal control was measured with flow cytometry. The entire coding regions of the CD18 gene were screened with direct sequencing genomic DNA.

RESULTSCD18 expression level on this patient's leukocyte surface was significantly decreased, with normal level in control group, his father and mother. Gene analysis revealed that this patient had a homozygous c.899A > T missense mutation in exon 8 of CD18 gene, causing the substitution of Asp to Val at the 300 amino acid. His parents were both heterozygous carriers while no such mutation was found in 50 normal controls.

CONCLUSIONThis study disclosed a novel point mutation Asp 300 Val located in a highly conserved region (HCR) of CD18 and confirmed the heterogeneity of the mutations causing LAD-1, indicating it was quite beneficial to establish correct and early diagnosis in children with severe LAD-1.

Asian Continental Ancestry Group ; CD18 Antigens ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Flow Cytometry ; Humans ; Leukocyte-Adhesion Deficiency Syndrome ; etiology ; genetics ; Male ; Point Mutation ; genetics ; Polymerase Chain Reaction

The aim of this study was to observe the change in angiotensin II receptor subtype 1 (AT(1)) autoantibody during the development of renovascular hypertension (RVH). The Goldblatt renovascular hypertension model was established by the two-kidney one-clip method, and a synthetic peptide corresponding to amino acid sequence 165-191 of the second extracellular loop of the AT(1)-receptor was used as the antigen. Sera AT(1)-receptor autoantibody was detected by SA-ELISA. It was shown that two weeks after operation both the frequency of occurrence and the titre of autoantibodies to AT(1)-receptor were significantly increased as compared with the pre-treatment control. The increase in autoantibodies lasted several weeks and then decreased gradually to the pre-clipping level at 12 weeks. It is suggested that autoimmune mechanisms are involved in the pathogenesis of renovascular hypertension and the AT(1) autoantibodies may be one of the mechanisms leading to cardiac hypertrophy.
Animals ; Autoantibodies ; blood ; Disease Models, Animal ; Hypertension, Renovascular ; blood ; immunology ; Kidney ; physiopathology ; Rats ; Receptor, Angiotensin, Type 1 ; immunology ; metabolism

AIM AND METHODSThe effects of losartan (after operation 2 week to 10 week, 5 mg/kg d ig) on generation of AT1R-AA in sera were observed during development of hypertension in rats. The renovascular hypertension (RVH) model was established by two-kidney one-clip method, a synthetic peptide corresponding to amino acid sequence 165-191 of the second extracellular loop of the angiotensin II-1 receptor (AT1R) was used as antigen, SA-ELISA were used to examine sera AT1R autoantibody (AT1R-AA).

RESULTSThe frequencies and titres of AT1R-AA after operation one week rats were significantly increased (P < 0.05). The treatment with losartan not only inhibited structural and functional changes, but also the frequencies and titres of AT1R-AA was significantly lower (P < 0.05) than RVH group.

CONCLUSIONIt is suggested that the losartan significantly inhibits generation of the AT1R-AA.

Animals ; Autoantibodies ; biosynthesis ; blood ; Hypertension, Renovascular ; blood ; immunology ; Losartan ; pharmacology ; Male ; Rats ; Rats, Wistar ; Receptors, Angiotensin ; immunology

Using two-kidney one-clip renal hypertensive (2K1C group), stress-induced hypertensive (neural group), DOCA-salt treated hypertensive (DOCA group) and spontaneously hypertensive rats (SHR group), to investigate the change in AT(1A)-receptor autoantibodies (AT(1A)-AAs) during the development of the four types of hypertension. The biological activities of AT(1A)-AAs were examined. It was shown that the frequency of occurrence and titres of AT(1A)-AAs increased significantly during the development of hypertension. In the four hypertensive groups studied, the occurrence of AT(1A)-AAs was most prominent in SHR, 2K1C and neural groups. The biological effects of AT(1A)-AAs were shown to increase the beating frequency of cultured neonatal myocardial and vascular contractile tension. It is suggested that autoimmune mechanisms are involved the pathogenesis of different types of hypertension and the AT(1A)-AAs may be one of the mechanisms leading to cardiac hypertrophy.
Animals ; Autoantibodies ; blood ; Desoxycorticosterone ; administration & dosage ; Hypertension ; classification ; etiology ; immunology ; physiopathology ; Hypertension, Renovascular ; immunology ; physiopathology ; Male ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; immunology ; Stress, Physiological ; physiology

OBJECTIVETo evaluate accuracy of Propex, Raypex 5, Root ZX electronic apex locator in positions of the perforation with different irrigations in the root canal.

METHODSPerforation lengths were measured with Propex, Raypex 5, Root ZX electronic apex locators in 19 extracted human teeth embedded in model after simulate perforation preparation by ultrasonic ET40 and compared with the actual canal length measurements taken before embedding the teeth in model. Measurements were taken with the different canal contents. RESULTS; Propex, Raypex 5, and Root ZX could locate the positions of the perforation with different irrigations in the root canal. The differences between actual root canal perforation length and measured root canal perforation length of same electronic apex locators in different irrigations were no statistically significant (P > 0.05). Meanwhile, the differences between actual root canal perforation length and measured root canal perforation length of three kinds of electronic apex locators in same irrigations were not statistically significant (P > 0.05).

CONCLUSIONPropex, Raypex 5, and Root ZX electronic apex locators can detect perforation accurately.

Dental Pulp Cavity ; Humans ; In Vitro Techniques ; Molar ; Odontometry ; Root Canal Therapy ; Tooth Apex ; Tooth Root

The study was designed to explore the alteration of intracellular calcium concentration ([Ca²⁺]i), induced by transient receptor potential melastatin 8 (TRPM8) channel-specific agonist menthol, in pulmonary arterial smooth muscle cells (PASMCs) between control and pulmonary hypertensive (PH) rats. PH rat models were established by means of chronic hypoxia (CH) and monocrotaline (MCT) injection, respectively. PASMCs from control and PH rats were cultured. The change of [Ca²⁺]i in PASMCs induced by menthol, and the effect of TRPM8 channel-specific antagonist BCTC on the change of [Ca²⁺]i, were observed. Cellular localization of TRPM8 was examined by using immunohistochemistry. Results showed that menthol increased [Ca²⁺]i in the control PASMCs both in Ca²⁺ -normal and Ca²⁺ - free Tyrode's solutions, and at the same time BCTC could inhibit these two kinds of elevations. Compared with the control group, elevations of [Ca²⁺]i were decreased notably in CH- and MCT-pretreated PASMCs superfused with 2 mmol/L Ca²⁺ - or 0 Ca²⁺ -Tyrode's solutions. Immunohistochemical localization experiments showed that the whole PASMCs were dyed brown except for the nucleus. This study verified that TRPM8 exists both in membrane and sarcoplasmic reticulum of PASMCs. In addition, CH- and MCT-pretreatment could independently down-regulate the Ca²⁺ influx and Ca²⁺ release mediated by TRPM8 channel.
Animals ; Calcium ; metabolism ; Cells, Cultured ; Menthol ; pharmacology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Pulmonary Artery ; cytology ; Rats ; Sarcoplasmic Reticulum ; metabolism ; TRPM Cation Channels ; metabolism