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Objective:To explore the effect and mechanism of Guizhitang with different proportions of Cinnamomi Ramulus and Paeoniae Alba Radix on overactive cardiac sympathetic nerves in salt-sensitive hypertensive rats. Method:Randomly divide the forty male 6-week-old salt-sensitive hypertensive rats into five groups： the normal control group， the model group， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶2 group，and the Cinnamomi Ramulus and Paeoniae Alba Radix 2∶1 group， each group has 8 animals， the normal control group was fed with low-salt feed， and the remaining four groups were fed with 8% high-salt feed. After 4 weeks of feeding， gastric feeding was started. Give both the normal control group and model group saline and the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group， the 1∶2 group，and the 2∶1 group， were given Guizhitang aqueous solution at 4.0， 5.5 and 5.5 g·kg-1， respectively. Continuous gavage intervention was held for 4 weeks. IITC multi-channel non-invasive sphygmomanometer was used to detect changes of systolic blood pressure before and after treatment in rats. Left ventricular anterior wall end-diastolic thickness （LVAWd） and interventricular septal diastolic thickness （IVSd） were detected by echocardiography. Hematoxylin-eosin（HE）staining and Masson staining were used to observe the myocardial morphological changes of rats in each group， Western blot was used to detect the expression of nerve growth factor （NGF）， growth-associated protein 43 （GAP43） and tyrosine hydroxylase （TH） protein. Result:After 4 weeks of intervention with Guizhitang， compared with the normal control group， the blood pressure， LVAWd and IVSd of the model group were significantly increased， and the expressions of NGF， TH and GAP43 protein were significantly increased （P<0.01）. HE and Masson staining results showed that the model group had myocardial cell edema， a large number of inflammatory cell infiltration， myocardial fiber hyperplasia and disordered arrangement， and a large amount of collagen deposition could be seen in the intercellular substance. Compared with model group， the systolic blood pressure of rats in each Guizhitang group increased slowly， and the expression of NGF， TH and GAP43 protein decreased （P<0.05，P<0.01），the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group has the best effect. The results of echocardiography shows that the 1∶1 Guishao group could reduce LVAWd and IVSd levels （P<0.05，P<0.01）， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶2 group could reduce IVSd level （P<0.05）， there was no statistical difference in LVAWd， there was no statistical difference in LVAWd and IVSd in 2∶1 group. In terms of myocardial morphology， each group of Guizhitang can reduce cell edema and inflammatory cell infiltration， reduce myocardial fiber hyperplasia and collagen deposition， and improve the disorder of myocardial fiber arrangement. Among them， the 1∶1 group has the best effect. Conclusion:Guizhitang can inhibit the overactive activation of cardiac sympathetic nervous system，reduce the extent of myocardial fibrosis， inflammatory infiltration and myocardial hypertrophy， and protect salt-sensitive hypertension rats， whose mechanism may be related to regulating the expression of heart NGF.Among them， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group is better than the 1∶2 and 2∶1 group in reducing myocardial fibrosis， inflammatory infiltration and myocardial hypertrophy.
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<b>Objective::To observe the effect of Guizhitang with different proportions of Cinnamomi Ramulus and Paeoniae Alba Radix on the expressions of transforming growth factor-<italic>β</italic><sub>1</sub>(TGF-<italic>β</italic><sub>1</sub>)/Smads signaling pathway and interleukin-10(IL-10), IL-6 and tumour necrosis factor-<italic>α</italic>(TNF-<italic>α</italic>)related inflammatory cytokines in salt-sensitive hypertensive rats, in order to explore the mechanism of Guizhitang in improving myocardial fibrosis in salt-sensitive hypertensive rats. <b>Method::Totally 40 male 6-week-old salt-sensitive rats were randomly divided into 5 groups: the normal control group, the model group, the 1∶1(RC/peony)Guishao group, the 1∶2 Guishao group, and the 2∶1 Guishao group, with 8 in each group. The normal control group was fed with normal salt diet, while the other four groups were fed with high-salt diet. After 4 weeks of feeding, the rats were given intragastric administration, the normal control group and the model group were given the same amount of normal saline, and the 1∶1 Guishao group, the 1∶2 Guishao group and the 2∶1 Guishao group were given 4.0, 5.5, 5.5 g·kg<sup>-1</sup> of Guizhitang by gavage for 4 weeks. Blood pressure was measured once a week, left ventricular end systolic diameter(LVESD), left ventricular end diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF) and left ventricular short axis shortening fraction (LVFS) were detected by using echocardiogram. The pathological changes of myocardial morphology were observed by htoxylin eosin(HE)and Masson staining. The expressions of type Ⅰ and Ⅲ collagen in myocardial tissue of each group was detected by immunohistochemistry. The mRNA expression levels of IL-10, IL-6 and TNF-<italic>α</italic> in myocardial tissue of each group were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR). The protein expression levels of TGF-<italic>β</italic><sub>1</sub>, <italic>α</italic>-smooth muscle actin(<italic>α</italic>-SMA), Smad2, Smad3 and Smad7 in myocardial tissue of each group were detected by Western blot. <b>Result::Compared with the normal control group, the blood pressure was increased in the model group at 8-15 weeks, LVESD, LVEDD were increased in the model group, while LVFS, LVEF were decreased in the model group. The collagen volume fraction was increased, immunohistochemistry showed the expression levels of type Ⅰ and Ⅲ collagen were increased, mRNA expression levels of IL-10, IL-6 and TNF-<italic>α</italic> were increased, the protein expression levels of TGF-<italic>β</italic><sub>1</sub>, Smad2, Smad3 and <italic>α</italic>-SMA were increased, whereas the protein expression of Smad7 was decreased (<italic>P</italic><0.01). Compared with the model group, the blood pressure rise of each group of Guizhitang was delayed in 12-15 weeks, LVESD and LVEDD were decreased in Guizhitang group (<italic>P</italic><0.01), LVFS, LVEF were increased in Guizhitang group (<italic>P</italic><0.01), the collagen volume fraction was decreased in Guizhitang group (<italic>P</italic><0.01), and the expressions of type Ⅰ and Ⅲ collagen were decreased in Guizhitang group (<italic>P</italic><0.01). At the same time, the mRNA expression of IL-10 was increased in Guizhitang group (<italic>P</italic><0.05, <italic>P</italic><0.01), the mRNA expressions of IL-6 and TNF-<italic>α</italic> were decreased in Guizhitang group (<italic>P</italic><0.01), and the protein expressions of TGF-<italic>β</italic><sub>1</sub>, Smad2, Smad3 and <italic>α</italic>-SMA were decreased in Guizhitang group (<italic>P</italic><0.05, <italic>P</italic><0.01), and the protein expression of Smad7 was increased in Guizhitang group (<italic>P</italic><0.01). Compared with the 2∶1 Guishao group, the effect of the 1∶1 Guishao group in improving the above indicators was more obvious (<italic>P</italic><0.05, <italic>P</italic><0.01). <b>Conclusion::Guizhitang with different proportions of Ramulus Cinnamomi and Poeny can alleviate the degree of myocardial fibrosis in salt-sensitive hypertensive rats. The mechanism may be related to the regulation of TGF-<italic>β</italic><sub>1</sub>/Smads signaling pathway and the reduction of inflammatory response. Besides, the 1∶1 Guishao group showed the optimal effect in reducing inflammation and improving myocardial fibrosis.
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Objective: To observe the expression and significance of fibronectin and metalloproteinase-3 (MMP-3) in patients with ankylosing spondylitis (AS). Methods: A total of 30 AS patients in our hospital and 30 healthy volunteers were selected in our study. Fibronectin and MMP-3 were measured and compared between these two groups. The AS group received sulfasalazine 2 g daily for 3 months. Bath ankylosing spondylitis disease activity index, bath ankylosing spondylitis functional index, bath ankylosing spondylitis metrology index, erythrocyte sedimentation rate and C-reactive protein were compared before and after treatment. Pearson's linear-correlation analysis was used to determine relationships between parameters. Results: Totally 28 patients in the AS group completed the study. Fibronectin and MMP-3 in peripheral blood of AS patients were evidently higher than that in the normal control group (P<0.05). After treated by sulfasalazine, the level of expressing Fibronectin and MMP-3 significantly decreased compared with baseline values (P<0.05). Pearson's linear-correlation analysis showed that serum fibronectin and MMP-3 level had a positive correlation with bath ankylosing spondylitis disease activity index global assessment, spine pain, night pain, general pain, erythrocyte sedimentation rate and C-reactive protein (P<0.05). Conclusions: The expression of fibronectin and MMP-3 in AS patients were significantly higher than that in the normal control group, and they all decreased significantly after treatment. It indicated that both fibronectin and MMP-3 were correlated closely with the onset of AS.
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Objective:To observe the expression and significance of fibronectin and metalloproteinase-3 (MMP-3) in patients with ankylosing spondylitis (AS).Methods:A total of 30 AS patients in our hospital and 30 healthy volunteers were selected in our study. Fibronectin and MMP-3 were measured and compared between these two groups. The AS group received sulfasalazine 2 g daily for 3 months. Bath ankylosing spondylitis disease activity index, bath ankylosing spondylitis functional index, bath ankylosing spondylitis metrology index, erythrocyte sedimentation rate and C-reactive protein were compared before and after treatment. Pearson's linear-correlation analysis was used to determine relationships between parameters.Results:Totally 28 patients in the AS group completed the study. Fibronectin and MMP-3 in peripheral blood of AS patients were evidently higher than that in the normal control group (P<0.05). After treated by sulfasalazine, the level of expressing Fibronectin and MMP-3 significantly decreased compared with baseline values (P<0.05). Pearson's linear-correlation analysis showed that serum fibronectin and MMP-3 level had a positive correlation with bath ankylosing spondylitis disease activity index global assessment, spine pain, night pain, general pain, erythrocyte sedimentation rate and C-reactive protein (P<0.05).Conclusions:The expression of fibronectin and MMP-3 in AS patients were significantly higher than that in the normal control group, and they all decreased significantly after treatment. It indicated that both fibronectin and MMP-3 were correlated closely with the onset of AS.
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Objective: To identify the authenticity of Bupleuri Radix in the market of Hebei province. Methods: In this research, the ITS2 sequence of 28 samples of Bupleuri Radix that were bought from the market in Hebei province was amplified by PCR, and the sequence of seven samples was downloaded from GenBank. Aligner CodonCode software was used for sequence alignment, the MEGA6.0 analysis was used to calculate the K2P distance, and the mutation sites of each sequence was analyzed. Results: Among the 28 Bupleuri Radix samples, there were 21 samples for Bupleurum chinese, 3 for B.smithii, and 4 for the false. Conclusion: ITS2 sequence can be used as an effective method to identify the authenticity of Bupleuri Radix, it is great value to the medicine market standard and the clinical medication safety.