Hypoglycemic Effect of Radix Et Rhizoma Salviae Miltiorrhizae,Guixi Green Tea and Their Compatibility on Mice

[Objectives]To explore the hypoglycemic effect of Radix Et Rhizoma Salviae Miltiorrhizae,Guixi green tea and their compatibility on mice,so as to provide more options for the adjuvant treatment of diabetes and improve the comprehensive utilization value of Guixi green tea.[Methods]After being fed adaptively for 7 d,60 SPF-grade male mice were intraperitoneally injected with alloxouracil(180 mg/kg)to induce hyperglycemia model,from which 35 mice with a successful model were selected.They were randomly divided into 5 groups(Danshen group,green tea group,compatible group,positive control group and blank control group),with 7 animals in each group.The mice in all groups were administered according to the dose of 200 mg/kg once a day for 15 d.After the last administration,their body weight was measured on the 15 th day after fasting for 12 h,and their fasting blood glucose was measured by tail clipping.[Results]After modeling,the mice in each group showed typical hyperglycemia symptoms,namely obvious polydipsia,polyphagia,polyuria and weight loss.Compared with the blank control group,Danshen group and green tea group had obvious hypoglycemic effect(P<0.05),and compatible group had significant hypoglycemic effect(P<0.01).Compared with the blank control group,the weight loss of diabetic mice could also be inhibited in green tea group and compatible group(P<0.05).[Conclusions]Danshen,Guixi green tea and their compatibility had the effect of lowering blood sugar,among which the compatible group had the most obvious effect.

The Effect of Soil Enzymes and Polysaccharides Secreted by the Roots of Salvia miltiorrhiza Bunge under Drought,High Temperature,and Nitrogen and Phosphorus Deficits

Root exudates serve as crucial mediators for information exchange between plants and soil,and are an important evolutionary mechanism for plants’adaptation to environmental changes.In this study,15 different abiotic stress models were established using various stress factors,including drought(D),high temperature(T),nitrogen deficiency(N),phosphorus deficiency(P),and their combinations.We investigated their effects on the seedling growth of Salvia miltiorrhiza Bunge and the activities of Solid-Urease(S-UE),Solid-Nitrite Reductase(S-NiR),Solid-Nitrate Reductase(S-NR),Solid-Phosphotransferase(S-PT),and Solid-Catalase(S-CAT),as well as the contents of polysaccharides in the culture medium.The results showed that the growth of S.miltiorrhiza was inhibited under 15 stress conditions.Among them,13 stress conditions increased the root-shoot ratio.These 15 stress conditions significantly reduced the activity of S-NR,two combinations significantly improved the activity of S-NIR,they were synergistic stresses of high temperature and nitrogen deficiency(TN),and synergistic stresses of drought and nitrogen deficiency(DN)(p<0.05).The activity of S-UE was significantly improved under N,D,T,synergistic stresses of drought and high temperature(DT),DN,synergistic stresses of drought and phosphorus deficiency(DP),and synergistic stresses of high temperature,nitrogen,and phosphorus deficiency(TNP)stress conditions(p<0.05).Most stress combinations reduced the activity of S-PT,but D and T significantly improved it.(p<0.05).The N,DN,and TN stress conditions significantly reduced S-CAT activity.The P,DT,and synergistic stresses of drought,high temperature,and phosphorus deficiency(DTP)significantly decreased the total polysaccharide content of the soil(p<0.05).The research suggested that abiotic stress hindered the growth of S.miltiorrhiza and altered the behavior of root secretion.Roots regulated the secretion of several substances in response to various abiotic stresses,including soil nitrogen cycle enzymes,phosphorus transport-related enzymes,and antioxidant enzymes.In conclusion,plants regulate the utilization of rhizosphere substances in response to abiotic stresses by modulating the exudation of soil enzymes and polysaccharides by the root system.At the same time,soil carbon sequestration was affected by the adverse environment,which restricted the input of organic matter into the soil.

Progress in research into the mechanism of Radix salviae miltiorrhizae in treatment of acute pancreatitis

BACKGROUND: Acute pancreatitis (AP) is a common acute abdomen with high mortality, multiple complications and complicated causes. There is no effective therapy for AP. Radix salviae miltiorrhizae (Danshen), a traditional herbal medicine, has a low price and a wide range of clinical applications. It is effective to promote blood flow, eliminate stagnancy, and relieve pain. It is also found to be effective in treating AP. We reviewed the progress in research into the mechanism of Radix salviae miltiorrhizae in treatment of AP. DATA SOURCES: An english-language literature search was conducted using MEDLINE (1988-2005) on Radix salviae miltiorrhizae (Danshen) and acute pancreatitis. RESULTS: The mechanisms of Danshen in the treatment of AP include improvement of microcirculatory disturbances; elimination of oxygen free radicals; modulation of the metabolism of lipid inflammatory mediator; and blocking of calcium inflow and prevention of calcium overload.CONCLUSION: Danshen can effectively reduce the mortality and complications of AP.

Experimental Study on the Preventive Mechanism of Salviae Miltiorrhizae Against Atherosclerosis in Rabbits Models

Summary: The preventive mechanism of salviae miltiorrhizae (SM) against experimental atherosclerosis (AS) in rabbits models was investigated. The experimental AS rabbit models were reproduced by feeding the high cholesterol diet. The changes of atherosclerotic plaques in normal group, model group and SM treated group were observed. The levels of serum TG, TC, HDL-C and LDL-C were determined. The immunohistochemistry was used to detect the expression of Bcl-2, Bax and IL-6 proteins in atherosclerotic plaques. The results showed that the level of serum TG in SM treated group was significantly lower than in model group (P<0.01). Immunohistochemistry revealed that the expression of Bcl-2, Bax and IL-6 in model group was significantly higher than in normal group. In the SM group, the expression of Bcl-2 protein was up-regulated and that of Bax was down-regulated. It was suggested that SM could inhibit formation of AS in experimental rabbits. To decrease the expression of Bax and increase the expression of Bcl-2 protein may be one of the mechanisms of SM against atherosclerosis.

Salviae miltiorrhizae ameliorates cirrhosis and portal hypertension by inhibiting nitric oxide in cirrhotic rats

OBJECTIVE: To determine the effect of Salviae miltiorrhizae on cirrhosis and portal hyperiension by inhibiting nitric oxide synthase type Ⅱ (NOSⅡ) in rats. METHODS: Real time RT-PCR was used to detect the expression of NOSII mRNA. The enzymatic activity of nitric oxide synthase and the circulating levels of nitric oxide (NO), systemic and portal hemodynamics, and quantification of cirrhosis were measured with highly sensitive methods. Traditional Chinese medicine was utilized to treat cirrhotic rats and the function of NO was evaluated. Double-blind method was applied in the experiment constantly. RESULTS: The concentration of NO increased markedly at all stages of cirrhosis, and so did the enzymatic activity of NOS, antl the iNOSmRNA expressed greatly. Meanwhile the portal-venous-pressure (PVP), portal-venous-now (PVF) increased significantly. NO, Nos and iNOSmHNA were positively correlated to the quanlity of hepatic fihrosis. Salviae Miltiorrhizae significantly inhibited NO production and inhibited the expression of iNOSmRNA. CONCLUSIONS: The increased hepatic expression of NoSIⅡ is one of the important factors causing cirrhosis and portal hypertension. Salviae Miltiorrhizae significantly ameliorates cirrhosis and portal hypertension.

Effect of salviae miltiorrhizae and ligustrazine hydrochloride injection on axonal regeneration and nerve growth factor expression in a rat model of sciatic nerve injury

BACKGROUND： Studies have shown that both salviae miltiorrhizae and ligustrazine can promote protein expression of nerve growth factor （NGF） and regeneration of peripheral nerve. OBJECTIVE： To verify the effect of salviae miltiorrhizae and ligustrazine hydrochloride injection on axonal regeneration and NGF protein expression in a rat model of sciatic nerve injury. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment was performed at the Laboratory of Traditional Chinese Medicine and the Institute of Bioengineering of Jinan University from July to December 2008. MATERIALS： Salviae miltiorrhizae and ligustrazine hydrochloride injection （containing 20 mg salviae miJtiorrhizae and 100 mg ligustrazine per 100 mL injection） was provided by Guizhou Baite Pharmaceutical, China; salviae miltiorrhizae and ligustrazine decoctions （containing 1 g raw drug per 1 mL decoction） were provided by Guangzhou Baiyunshan Factory for Traditional Chinese Medicine, China; rabbit-anti-rat NGF monoclonal antibody was provided by Beijing Biosynthesis Biotechnology, China. METHODS： A total of 80 healthy, male, Sprague Dawley rats were used to establish a sciatic nerve injury model via neurotomy, and were then randomly assigned to 4 groups： salviae miltiorrhizae and ligustrazine hydrochloride injection group （intraperitoneal injection of 35 mL/kg per day salviae miltiorrhizae and ligustrazine hydrochloride injection）, saIviae miltiorrhizae group （intragastric peffusion of 2 mL salviae miltiorrhizae）, ligustrazine group （intragastric peffusion of 2 mL ligustrazine）, and model group （intraperitoneal injection of 35 mL/kg per day saline）, with 20 rats in each group. Thereafter, rats in each group were then divided into 4 subgroups according to varying time points of 1, 2, 4, and 8 weeks post-surgery, with 5 rats in each subgroup. MAIN OUTCOME MEASURES： Axons were quantified using chromotrope 2R-brilliant green and silver staining combined with image analysis to calculate the axonal regeneration rate; NGF expression was detected using immunohistochemistry and Western blot analysis; toe interspace was measured by behavior at 4 and 8 weeks. RESULTS： With increasing time after sciatic nerve expression, and toe interspace gradually increased njury, the axonal regeneration rate, NGF protein At 4 and 8 weeks post-surgery, axonal regeneration rate and NGF protein expression were significantly increased in the injured tissue of the salviae miltiorrhizae and ligustrazine hydrochloride injection, salviae miltiorrhizae, and ligustrazine groups, compared with the model group （P 〈 0.05 or P 〈 0.01）, and toe interspace was remarkably enlarged （P 〈 0.05 or P 〈 0.01）, especially in the salviae miltiorrhizae and ligustrazine hydrochloride injection group. CONCLUSION： Salviae miltiorrhizae and ligustrazine hydrochloride injection promoted axonal regeneration and NGF protein expression in the injured sciatic nerve, and also enhanced neurofunctional recovery. Its effect was superior to salviae miltiorrhizae or ligustrazine alone.

Clinical study of adjuvant therapy on ischemic stroke with salviae miltiorrhizae and ligustrazine

Objective: To study the effect of adjuvant therapy with salviae miltiorrhizae and ligustrazine injection on neural function injury, degree of oxidative stress reaction, chemotactic factor and adhesion molecule in patients with ischemic stroke. <br> Methods: Patients with ischemic stroke admitted in our hospital from August 2013 to October 2015 were retrospectively analyzed and divided into conventional treatment (CT) group receiving conventional treatment and traditional Chinese medicine auxiliary (TCMA) group receiving adjuvant therapy with salviae miltiorrhizae and ligustrazine injection. After the treatment of the first month and the third month, serum was collected to detect the contents of nerve injury molecules, oxidative stress parameters, chemotactic factor and adhesion molecule. <br> Results: After the 4-week treatment, the contents of serum neuron specific enolase, S100 calcium-binding protein B, heart-type fatty acid binding protein, malondialdehyde, advanced oxidation protein products, 8-hydroxy-2'-deoxyguanosine, monocyte chemoattractant protein-1,CD40,CD40 ligand, vascular endothelial cadherin, soluble intercellular adhesion molecule 1 and soluble vascular adhesion molecule 1 in patients ofTCMA group were all significantly lower than those of CT group, and the contents of superoxidase dismutase, glutathione peroxidase and catalase were significantly higher than those of CT group. After the 8-week treatment, the contents of serum neuron specific enolase, S100 calcium-binding protein B, heart-type fatty acid binding protein, malondialdehyde, advanced oxidation protein products, 8-hydroxy-2'-deoxyguanosine, monocyte chemoattractant protein-1,CD40, CD40 ligand, vascular endothelial cadherin, soluble intercellular adhesion molecule 1 and soluble vascular adhesion molecule 1 in patients ofTCMA group were all significantly lower than those of CT group and the contents of superoxidase dismutase, glutathione peroxidase and catalase were significantly higher than those of CT group. <br> Conclusions: Adjuvant therapy with salviae miltiorrhizae and ligustrazine injection can alleviate the neural function injury, inhibit oxidative stress reaction and the generation of chemotactic factor and adhesion molecule in patients with ischemic stroke, which is an effective medicine for treating ischemic stroke.

Gastric mucosal injury due to hemorrhagic reperfusion and efficacy of Salvia miltiorrhizae extract F and cimetidine

AIM: To observe the gastric mucosal injury caused by hemorrhagic shock and reperfusion and to compare the effect between Salvia miltiorrhizae extract F (SEF) and cimetidine (CI) on it.METHODS: A model of hemorrhage/reperfusion injury was produced by Itoh method. Wistar rats were randomly divided into three groups: 0.9% sodium chloride treatment group (NS group), SEF treatment group (SEF group), and CI treatment group (CI group). Saline, SEF and CI were injected respectively. The index of gastric mucosal lesions (IGML) was expressed as the percentage of lesion area in the gastric mucosa. The degree of gastric mucosal lesions was categorized into grades 0, 1, 2, 3. Atom absorption method was used to measure the intracellular calcium content. Radioimmunoassay was used to measure the concentrations of prostaglandins.RESULTS: IGML (%) and grade 3 (%) were 23.18±6.82, 58.44±9.07 in NS group, 4.42±1.39, 20.32±6.95 in SEF group and 3.74±1.56, 23.12±5.09 in CI group, and theabove parameters in SEF group and CI group decreased significantly (IGML: SEF vs NS, t = 6.712, P = 0.000＜0.01; CI vs NS, t= 6.943, P= 0.000＜0.01; grade 3: SEF vs NS,t= 8.386, P= 0.000; CI vs NS, t= 8.411, P= 0.000), but the grade 0 and grade 1 damage in SEF group (22.05±5.96, 34.12±8.12) and CI group (18.54±4.82, 30.15±7.12) were markedly higher than those in NS group (3.01±1.01, 8.35±1.95; grade 0: SEF vs NS, t= 8.434,P = 0.000＜0.01; CI vs NS, t = 7.950, P = 0.000＜0.01;grade 1: SEF vs NS, t= 8.422, P= 0.000＜0.01; CI vs NS,t = 8.448, P = 0.000＜0.01). The intracellular calcium content (μg/mg) in SEF group (0.104±0.015) and CI group (0.102±0.010) was markedly lower than that in NS group(0.131±0.019, SEF vs NS, t= 2.463, P= 0.038＜0.05; CIvs NS, t= 3.056, P= 0.017＜0.05). The levels (pg/mg) ofPGE2, 6-keto-PGF1αand 6-keto-PGF1α/TXB2 were 540±183, 714±124, 17.38±5.93 in NS group and 581±168, 737±102, 19.04±8.03 in CI group, 760±192, 1 248±158, 33.42±9.24 in SEF group, and the above parameters in SEF group markedly raised (PGE2: SEF vs NS, t = 2.282, P = 0.046 ＜0.05; SEF vs CI, t = 2.265, P = 0.047＜0.05; 6-ketoPGF1α: SEF vsNS, t= 6.583, P= 0.000＜0.000; SEF vsCI,t = 6.708, P = 0.000＜0.01; 6-keto-PGF1α/TXB2: SEF vs NS, t= 3.963, P= 0.003＜0.001; SEF vs CI, t= 3.243,P = 0.009＜0.01), whereas TXB2 level in SEF group (45.37±7.54) was obviously lower than that in NS group (58.28±6.74, t = 3.086, P = 0.014＜0.05) and CI group (54.32±6.89, t = 2.265, P = 0.047＜0.05). No significant difference was shown between NS group and CI group (PGE2: t= 0.414, P= 0.688＞0.05; 6-keto-PGF1α: t= 0.310,P = 0.763＞0.05; TXB2: t = 1.099, P = 0.298＞0.05; 6-keto-PGF1α/TXB2: t = 0.372, P = 0.718＞0.05).CONCLUSION: Both SEF and CI could inhibit reperfusion induced injury in gastric mucosa, but with different mechanisms. SEF could not only enhance the protective effect of gastric rnucosa, but also abate the injury factors, while CI can only abate the injury factors.

Simultaneous Determination of Four Phenolic Acids in Radix Salviae Miltiorrhizae Formula Granules by QAMS

[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenolic acids,salvianolic acid B,tanshinol,lithospermic acid and rosmarinic acid in Radix Salviae Miltiorrhizae by HPLC and conduct methodological investigation.[Methods]Salvianolic acid B,with moderate retention time,good resolution,great response value and easy availability was selected as the internal standard,and the relative retention values and relative correction factors between tanshinol,rosmarinic acid,lithospermic acid and salvianolic acid B were established.The results of QAMS were compared with those of external standard method to verify the accuracy and applicability of QAMS.[Results]There were no significant differences between the calculated values of QAMS and the measured values of external standard method.The relative deviation was less than 3%,and the relative correction factors obtained from the experiment were credible.[Conclusions]QAMS can be used to determine the contents of phenolic acids in Radix Salviae Miltiorrhizae formula granules.

The Inhibitory Effect of Radix Salviae Miltiorrhizae on Hypoxic Structural Remodeling of Intra－acinarPulmonary Arteries

The inhibitory effect of Radix Salviae Miltiorrhizae (RSM) on hypoxic structural remodeling of intra-acinar pulmonary arteries (IAPA) was observed by light and electron microscopy and morphometry. It was found that RSM can not only dilate IAPA and relieve the hypoxic injuries to endothelia cells,but also inhibit the active muscularization of IAPA in the hypoxic animals, suggesting that RSM plays a very important role in inhibiting structural remodeling of IAPA and pulmonary hypertension.

Study on Optimization of Ethanol Reflux Extraction of Phenolic Acids from Salvia miltiorrhiza

The extraction technology of phenolic acid compounds from Salvia miltiorrhiza by ethanol reflux was studied. In this experiment, salvianolic acid B standard was used to make the standard curve. Single factor experiment and orthogonal experiment were used to study the extraction of different ethanol concentrations, reflux times and material-to-liquid ratios. The OD value of salvianolic acid compounds was measured with a spectrophotometer. The extraction rate of phenolic acid compounds under different extraction conditions was calculated through a regression equation, so as to obtain the optimal conditions for the ethanol reflux extraction process of Salvia miltiorrhiza. The experimental data can provide a reference for the ethanol reflux extraction process of salvianolic acids in the industry. According to the experiment, the extraction rate of phenolic acids in Salvia miltiorrhiza was the highest when the ethanol concentration was 60%, the reflux time was 1.5 hours, and the ratio of material-to-liquid was 1:10.

EFFECT OF RADIX SALVIAE MILTIORRHIZAE ON GROWTH OF ISOLATED CELLS FROM EMBRYONIC CHICKEN FRONTAL BONE CULTURED IN VITRO (A HISTOCHEMICAL STUDY) Ⅱ.THE DEVELOPMENT AND MATURATION OF OSTEOBLAST-LIKE CELLS

The isolated osteoblast-like cells from embryonic chicken frontal bone werecultured in vitro and histochemical methods adopted to observe the effect of RadixSalviac Miltiorrhizae (RSM) on proliferation, differentiation, and osteogenic capacity ofthese cells. It was found that: 1. The mitosis and proliferation of the osteoblast-like cellscould be accelerated by RSM, resulting in increased density of the cells in RSM groupas compared with the control. 2. After 48 h, the pseudopodia stretched out and drew backactively in osteoblast-like cells in RSM group. Small particles produced in the cells weresecreted through exocytosis to the extracellular medium. However, in the control group,the capacity to form and secrete these particles was limited. These particles showed posi-tive Alcian blue staining in Alcian blue-Sirius red reaction, so they were acidmucopolysaccharide particles. 3. The osteoblast-like cells could secrete vesicular particles 3micra in diameter. These vesicular particles could be stained with Alcian blue in earlystage, then they could be stained with Sirius red, and finally by Alizarin red S. Thesevesicular particles could aggregate and fuse around the cell colonies, forming bonenodules and bone flakes. The quantity and volume of the bone nodules and flakes inRSM group were larger than in the control group. 4. The bone nodules and flakes couldbe labeled vitally with tetracycline, and show strong yellow fluorescence under thefluorescence microscope. Therefore, these substances were the newly formed bone sub-stances.

EFFECT OF RADIX SALVIAE MILTIORRHIZAE ON THE PROCOAGULANT AND FIBRINOLYTIC ACTIVITIES INDUCED BY ENDOTOXIN-TREATED BOVINE ENDOTHELIAL CELLS

Effect of endotoxin, and endotoxin plus Radix Salviae Miltiorrhizae （RSM） on bovine aortic endothelial cells （BAECs） for the production of tissue factor （TF）, plasminogen activator （PA） and plasminogen activator inhibitor （PAI） activities as well as prostacyclin （PGI2） content were studied. Stimulation of BAECs with endotoxin （1 μg/ ml） increased the expression of cell surface TF activity, the secretions of PAI activity and PGI2 content. However, PA activity in endotoxin-treated groups didn’t change significantly in comparison with that of the control. RSM could stimulate the secretion of PA activity and prevent the increase in TF and PAI activities from endotoxin-treated BAECs. However, there was no significant difference in PGI2 production between endotoxin-and endotoxin+RSM-treated BAECs. These results indicated that increased procoagulant function and decreased fibrinolytic activity of endothelial cells （ECs） might be one of the mechanisms of DIC in gram-negative septicemia and RSM would be a very important thug in the prevention and the treatment of DIC in gram-negative sepsis.

EFFECTS OF RADIX SALVIAE MILTIORRHIZAE AND ITS COMPONENT "DANSHENSU" ON THE PRODUCTION OF PA, PAI, PGI_2 AND EXPRESSION OF THROMBOMODULIN BY BOVINE ENDOTHELIAL CELLS IN CULTURE

The effects of Radix Salviae Miltiorrhizae and its component "DANSHENSU" on the production of PA, PAI, PGI2 and expression of thrombomodulin by cultured bovine endothelial cells were studied. 6-Keto-PGF1α was measured with RIA. PA, PAI and thrombomodulin were measured with chromosenic substrate S2390 and S2238 respectively. The results showed that Radix Salviae Miltiorrhizae could promote PA activity and PGI2 production by bovine endothelial cell （BEC）. It could inhibt activity of PAI secreted by BEC. Its component "DANSHENSU" had the same effects. In addition, Radix Salviae Miltiorrhizae could also increase thrombomodulin activity on the surface of BEC, but "DANSHENSU" did not.

THE EFFECT OF RADIX SALVIAE MILTIORRHIZAE ON THE INHIBITORY ACTIVITY OF VASCULAR SMOOTH MUSCLE CELLS TO THE PLASMINOGEN ACTIVATORS SECRETED BY ENDOTHELIAL CELLS

Cultured porcine endothelial cells (EC) produce and secrete plasminogenactivators (PA). If the serum free media incubated by vascular smooth muscle cells(SMC-CM) were mixed with the same media incubated by endothelial cells (EC-CM),the PA activities of the latter decreased significantly. Cocultivation of EC with SMC alsoresulted in a significant decrease (70.7%) of PA activities produced by EC. Sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of SMC-CMfollowed by reverse fibrin autography demonstrated that the PA inhibitor had a molecularweight of 49000-62000. In this study we also investigated the effect of a Chinese herbalmedicine-Radix Salviae Miltiorrhizae (RSM) on the inhibitory activity of SMC. The re-sults showed that RSM significantly decreased the inhibitory activity of SMC against thePA secreted by EC.

Protective effect of ultrashortwave versus radix salviae miltiorrhizae on brains of rats with cerebral ischemia-reperfusion injury

BACKGROUND： HOW to control the effect of oxygen-derived free radicals on development of cerebral injury and cerebral edema is a key factor for treating cerebral ischemia-reperfusion injury. OBJECTIVE： To observe and compare the protective effects, synergistic action and mechanisms of ultrashortwave （USW） and radix salviae miltiorrhizae （RSM） on the focal cerebral ischemia-reperfusion injuries in rats. DESIGN： Randomized controlled animal study SEI-FING： Department of Rehabilitation Medicine, First Hospital affiliated to China Medical University MATERIALS： A total of 160 healthy Wistar rats of both genders and aged 18-20 weeks weighing 250-300 g of clean grade were selected in this study. 5 mL/ampoule RSM injection fluid was produced by the First Pharmaceutical Corporation of Shanghai （batch number： 011019, 0.01 mug）. The USW therapeutic device was produced by Shanghai Electronic Device Factory with the frequency of 40.68 MHz and the maximal export power of 40 W. The first channel of power after modulation was 11 W. METHODS： The experiment was carried out in the Rehabilitation Medicine Department of the First Hospital affiliated to China Medical University from May 2002 to January 2003. Focal ischemia-reperfusion model was established in rats by reversible right middle cerebral artery occlusion with filament. Right cerebral ischemia was for 2 hours and then with 24 hours reperfusion. The scores of neurological deficits were evaluated by 0 to 4 scales. After surgery, 64 successful rats models were divided into four groups according to digital table： control group, USW group, RSM group and RSM ＋ USW group with 16 cases in each group. Rats in control group were intraperitoneally injected with the same volume of saline （0.1 mL/g）; rats in USW group were given small dosage of USW on head for 10 minutes at 6 hours after reperfusion; rats in RSM group were intraperitoneally injected with 0.01 mL/g RSM solution at 30 minutes before reperfusion; rats in RSM ＋ USW group were intraperitoneally injected with 0.01 mL/g RSM parenteral solution at 30 minutes before reperfusion and given small dosage of USW on head for 10 minutes once at 6 hours after reperfusion; sixteen rats in sham operation group did not receive any treatment. All 80 rats were taken brains at 24 hours after reperfusion to measure wet and dry weights to calculate water content： Cerebral water content （%） = （1-dry/wet weight） × 100%. Superoxide dismutase （SOD） activity was measured by hydroxylamine method and malondialdehyde （MDA） content was measured by TBA photometric method. MAIN OUTCOME MEASURES ： Cerebral water content, SOD activity and MDA content RESULTS： All 160 rats except 80 failing in modeling were involved in the final analysis. （① The cerebral water content of left hemisphere made no significant difference （P 〉 0.05）. The cerebral water content of right hemisphere in the control group and the three treatment groups was obviously higher than that of the sham operation group [（81.26±0.77）%, （79.74±0.68）%, （79.76±0.81）%, （79.61±0.79）%, （77.43±0.61）%, P 〈 0.05]. The cerebral water content of right hemisphere in the three treatment groups was obviously lower than that of the control group （P〈 0.05）. There was no significant difference among the three treatment groups （P 〉 0.05）. ② Compared with the control group, SOD activity （right） of the control group decreased obviously （P 〈 0.05）, while MDA content increased obviously （P 〈 0.05）. SOD activity in the three therapeutic groups increased obviously, while MDA content decreased obviously （P 〈 0.05）; there was no significant difference among the three treatment groups （P 〉 0.05）. CONCLUSION： ① USW and RSM therapy have neuroprotective effects against focal cerebral ischemia-reperfusion injuries by means of decreasing cerebral water content and MDA and increasing the activity of SOD. ② Synergistic action was not observed between these two therapeutic methods.

Characteristics of Salvia miltiorrhiza methylome and the regulatory mechanism of DNA methylation in tanshinone biosynthesis

Salvia miltiorrhiza is a model medicinal plant with significant economic and medicinal value.Its roots produce a group of diterpenoid lipophilic bioactive components,termed tanshinones.Biosynthesis and regulation of tanshinones has attracted widespread interest.However,the methylome of S.miltiorrhiza has not been analysed and the regulatory mechanism of DNA methylation in tanshinone production is largely unknown.Here we report single-base resolution DNA methylomes from roots and leaves.Comparative analysis revealed differential methylation patterns for CG,CHG,and CHH contexts and the association between DNA methylation and the expression of genes and small RNAs.Lowly methylated genes always had higher expression levels and 24-nucleotide sRNAs could be key players in the RdDM pathway in S.miltiorrhiza.DNA methylation variation analysis showed that CHH methylation contributed mostly to the difference.Go enrichment analysis showed that diterpenoid biosynthetic process was significantly enriched for genes with downstream overlapping with hypoCHHDMR in July_root when comparing with those in March_root.Tanshinone biosynthesis-related enzyme genes,such as DXS2,CMK,IDI1,HMGR2,DXR,MDS,CYP76AH1,2OGD25,and CYP71D373,were less CHH methylated in gene promoters or downstream regions in roots collected in July than those collected in March.Consistently,gene expression was up-regulated in S.miltiorrhiza roots collected in July compared with March and the treatment of DNA methylation inhibitor 5-azacytidine significantly promoted tanshinone production.It suggests that DNA methylation plays a significant regulatory role in tanshinone biosynthesis in S.miltiorrhiza through changing the levels of CHH methylation in promoters or downstreams of key enzyme genes.

Chemical constituents and pharmacological activities of Salvia miltiorrhiza

Salvia miltiorrhiza Bunge(S.miltiorrhiza),a perennial plant of the genus Salvia,is widely used in traditional folklore medicine.Previous chemical research on this plant contains diterpenoid quinones,phenolic acids,polysaccharides and other compounds.The pharmacological investigation of S.miltiorrhiza has shown that it has various pharmacological activities,such as cardiovascular system protection,anti-inflammatory,anti-oxidant,anti-tumor,liver protection,and neuroprotection activities.This research tends to give an overview of the main chemical constituents and pharmacological effects of S.miltiorrhiza,aiming to reveal its potential value and provide reference for its further development.

TREATMENT OF 40 CASES OF CHRONIC ECZEMA WITH POINT-INJECTION OF COMPOUND RADIX SALVIAE MILTIORRHIZAE INJECTION

The authors have used point-injection of compound Radix Salviae Miltiorrhizae Injection to treat 40 cases of chronic eczema and achieved a satisfactory therapeutic effect. The total effective rate was 95%.

不同垄作方式和种植密度对丹参产量与主要成分含量的影响

为探究不同栽培方式和生长时期对丹参根与地上部产量和主要活性成分含量的影响,本研究以紫花丹参为试验材料,采用小垄单行[三种密度:H1(8万株·hm^(-2))、H2(10万株·hm^(-2))、H3(13.3万株·hm^(-2))]与大垄双行[三种密度:H4(8万株·hm^(-2))、H5(10万株·hm^(-2))、H6(13.3万株·hm^(-2))]两种垄作方式,动态取样,测定并分析不同垄作方式、种植密度和生长时期的单株重、产量、主要活性成分含量及积累规律。结果表明,大垄双行丹参单株干重、产量、根的主要活性成分含量平均值均高于小垄单行。根的单株干重和产量的最佳处理分别为H5(48.09 g)、H4(5986.69 kg·hm^(-2)),均在11月19日最高;地上部单株干重和产量的最佳处理均为H4(50.34 g、6712.75 kg·hm^(-2)),均在10月14日最高。根中主要活性成分总含量(总丹参酮+总黄酮+总酚酸)在两种垄作方式下均呈“升-降-升”的变化趋势,最佳处理为H4(295.4 mg·g^(-1)),在9月7日最高;叶、茎中(总黄酮+总酚酸)总含量的最佳处理分别为H4(238.42 mg·g^(-1))、H6(122.43 mg·g^(-1)),均在10月14日最高;丹参酮ⅡA、丹参酮I、隐丹参酮和丹酚酸B四种活性成分总含量的最佳处理为H6(63.23 mg·g^(-1))。综上,大垄双行栽培方式优于小垄单行。本研究结果为丹参的合理种植及根与地上部综合开发利用提供了理论依据。