Rehmanniae Radix Praeparata Improves Neurological Function of Ischemic Stroke Rats by Inhibiting Autophagy and Ferroptosis / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Rehmanniae Radix Praeparata on neurological function injury in ischemic stroke rats and explore its mechanism. MethodMale SD rats were randomized into sham operation， model， low- and high -dose （3.5 g·kg-1 and 7 g·kg-1） Rehmannia Radix Praeparata， and nimodipine （0.01 g·kg-1） groups. The rat model of middle cerebral artery occlusion （MCAO） was established with the modified suture occlusion method. Zea-Longa 5-point scoring was employed to evaluate the neurological function of rats. The cerebral infarction volume was detected by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Hematoxylin-eosin staining and Nissl staining were employed to observe the morphology and damage of the brain tissue. Meanwhile， the serum levels of lactate dehydrogenase （LDH）， oxidative stress-related indicators superoxide dismutase （SOD）， glutathione peroxidase 4 （GPX4）， and malondialdehyde （MDA）， and the iron （Fe） content in the brain tissue were determined. To explore the mechanism of Rehmanniae Radix Preparata in mitigating the neurological damage in ischemic stroke rats， Western blotting was employed to determine the expression levels of proteins in the ischemic brain tissue. The autophagy-associated proteins included autophagy effector （beclin-1）， microtubule-associated protein light chain 3 （LC3B）， and ubiquitin-binding protein p62 （p62）. The ferroptosis-associated proteins included transferrin （TF）， transferrin receptor 1 （TFR1）， ferritin heavy chain 1 （FTH1）， and ferropotin （FPN1）. The neurological function injury-associated proteins included brain-derived neurotrophic factor （BDNF） and tyrosine kinase receptor B （TrkB）. ResultCompared with the sham operation group， the model group showed increased neurological function score， cerebral infarction volume， and appearance of nuclear pyknosis and vacuole of cells in the cerebral cortex. In addition， the model group presented elevated levels of LDH， MDA， and Fe （P<0.01） and lowered levels of SOD and GPX4 （P<0.01）. Compared with the model group， Rehmanniae Radix Praeparata decreased the content of LDH， MDA， and Fe （P<0.05， P<0.01） and elevated the levels of SOD and GPX4 （P<0.05， P<0.01）. Compared with the sham operation group， the modeling promoted the expression of beclin-1，LC3B Ⅱ/Ⅰ， TF， and TFR1 and inhibited the expression of p62， FTH1， FPN1， BDNF， and TrkB （P<0.01）. The expression levels of these proteins were recovered after the treatment with Rehmanniae Radix Praeparata. ConclusionRehmanniae Radix Praeparata may inhibit ferroptosis and improve the neurological function in ischemic stroke rats by down-regulating the autophagy level in the brain tissue.

2,3-Seco and 3-nor guaianolides fromAchillea alpina with antidiabetic activity / 中国天然药物

In this study, we presented the isolation and characterization of eight novel seco-guaianolide sesquiterpenoids (1-8) and two known guaianolide derivatives (9 and 10), from the aerial part of Achillea alpina L.. Compounds 1-3 were identified as guaianolides bearing an oxygen insertion at the 2, 3 position, while compounds 4-8 belonged to a group of special 3-nor guaianolide sesquiterpenoids. The structural elucidation of 1-8, including their absolute configurations, were accomplished by a combination of spectroscopic data analysis and quantum electronic circular dichroism (ECD) calculations. To evaluate the potential antidiabetic activity of compounds 1-10, we investigated their effects on glucose consumption in palmitic acid (PA)-mediated HepG2-insulin resistance (IR) cells. Among the tested compounds, compound 7 demonstrated the most pronounced ability to reverse IR. Moreover, a mechanistic investigation revealed that compound 7 exerted its antidiabetic effect by reducing the production of the pro-inflammatory cytokine IL-1β, which was achieved through the suppression of the NLRP3 pathway.

Ephedra Herb extract ameliorates adriamycin-induced nephrotic syndrome in rats via the CAMKK2/AMPK/mTOR signaling pathway / 中国天然药物

This study aimed to investigate the effect and mechanisms of Ephedra Herb (EH) extract on adriamycin-induced nephrotic syndrome (NS), providing an experimental basis for the clinical treatment of NS. Hematoxylin and eosin staining, creatinine, urea nitrogen, and kidn injury molecule-1 were used to evaluate the activities of EH extract on renal function. The levels of inflammatory factors and oxidative stress were detected by kits. The levels of reactive oxygen species, immune cells, and apoptosis were measured by flow cytometry. A network pharmacological approach was used to predict the potential targets and mechanisms of EH extract in the treatment of NS. The protein levels of apoptosis-related proteins and CAMKK2, p-CAMKK2, AMPK, p-AMPK, mTOR and p-mTOR in the kidneys were detected by Western blot. The effective material basis of EH extract was screened by MTT assay. The AMPK pathway inhibitor (compound C, CC) was added to investigate the effect of the potent material basis on adriamycin-induced cell injury. EH extract significantly improved renal injury and relieve inflammation, oxidative stress, and apoptosis in rats. Network pharmacology and Western blot results showed that the effect of EH extract on NS may be associated with the CAMKK2/AMPK/mTOR signaling pathway. Moreover, methylephedrine significantly ameliorated adriamycin-induced NRK-52e cell injury. Methylephedrine also significantly improved the phosphorylation of AMPK and mTOR, which were blocked by CC. In sum, EH extract may ameliorate renal injury via the CAMKK2/AMPK/mTOR signaling pathway. Moreover, methylephedrine may be one of the material bases of EH extract.

Comparison of liposoluble constituents and study on in vitro antioxidant activities of different processed products of Huaizhong No. 1 Rehmannia glutionsa / 中国药房

OBJECTIVE To co mpare the difference of liposoluble constitue nts in different processed products of Huaizhong No.1 Rehmannia glutionsa （fresh R. glutionsa ，R. glutionsa and prepared R. glutionsa ），and to evaluate its in vitro antioxidant activity preliminarily. METHODS Liposoluble extracts were extracted from 3 processed products of R. glutionsa by Soxhlet extraction. Their constituents were analyzed by gas chromatography-mass spectrometry. The spectral library of NIST 98 system was used to automatically retrieve the mass spectrum information of components ，and the structures of compounds were identified in combination with relevant literature and by comparing with eight peak index and EPA/NIH library. Relative contents of the components were calculated by using peak area normalization method with Hewlett Packard software. The antioxidant activities of liposoluble constituents in 3 processed products of R. glutionsa were investigated by 1，1-diphenyl-2-picrylhydrazyl（DPPH）free radical scavenging. RESULTS A total of 79 liposoluble components were identified from different processed products of R. glutionsa，and 48，52 and 37 liposoluble compounds were identified from fresh R. glutionsa ，R. glutionsa and prepared R. glutionsa，respectively；their relative contents accounted for 92.69％，86.29％，92.89％ of the total components respectively. Among them ，there were 20 liposoluble compounds totally ，and their relative contents accounted for 88.73％，80.89％ and 85.87％ of liposoluble components in each processed product respectively ；they were mainly composed of fatty acids such as methyl linoleate，methyl palmitate and methyl oleate. In addition ，there were 18 unique liposoluble components in fresh R. glutionsa ， mostly terpenoids ；there were 17 and 6 unique liposoluble components in R. glutionsa and prepared R. glutionsa ，mostly alkanes. The results of antioxidant experiment showed that median scavenging concentrations of liposoluble components to DPPH limeng free radical were 0.756，0.660，0.758 mg/mL，respectively. CONCLUSIONS The common liposoluble components in different processed products of R. glutionsa are mostly acids；the unique liposoluble components in fresh R. glutionsa are mostly terpenoids ，and those of R. glutionsa and prepared R. glutionsa are mostly alkanes ；the liposoluble constituents possess in vitro antioxidant activities.

Ascending， Descending， Floating and Sinking Properties of Descurainiae Semen Lepidii Semen Based on Pulmonary Edema Model / 中国实验方剂学杂志

ObjectiveThe pulmonary edema （PE） model where the disease was located in the viscera was established according to the treatment principle that patients with the disease location inside should be treated with descending and sinking medicine， combined with changes in the disease tendency， to verify the scientificity of descending and sinking properties of Descurainiae Semen Lepidii Semen （SD）， and to preliminarily elucidate the scientific connotation of descending， ascending， floating and sinking of Chinese medicine. MethodSixty male SD rats were randomly divided into normal control group， model group （20 mg·kg-1）， positive drug group （dexamethasone， 0.075 mg·kg-1） and SD low （1.167 g·kg-1）， medium （2.334 g·kg-1）and high （4.668 g·kg-1） dose groups. The PE model was established by intrapleural injection of 1% carrageenan （2 mL·kg-1）. The evaluation indexes （lung autopsy， amount of pleural effusion， number of white blood cells， lung wet/dry weight ratio， lung water content and lung permeability） were tested to determine the optimal dose of SD decoction for intervention of PE. The relevant indexes of the five major systems （central nervous system， respiratory system， circulatory system， digestive system and urinary system） closely related to the body's Qi movement were detected and changes in the disease tendency in PE rats were analyzed， to determine the descending， ascending， floating and sinking properties of SD. In addition， histopathological changes were investigated by hematoxylin-eosin （HE） staining， and types and numbers of inflammatory cells and mediators were detected to preliminarily explore the mechanism of SD in improving PE. ResultCompared with the conditions in the normal control group， the amount of pleural effusion， number of white blood cells in pleural effusion， lung wet/dry weight ratio， lung water content and lung permeability were increased （P<0.01） in the model group， where the rats presented cough， dyspnea， shortness of breath and arched back， and a small number of them had wet nose and bubble liquid in nostrils. In the autopsy of the rats in the model group， the lungs were enlarged or accompanied by congestion and plenty of pink bubble liquid appeared at the trachea. Compared with the conditions in the model group， SD reduced the amount of pleural effusion， number of white blood cells in pleural effusion， lung coefficient， lung wet/dry weight ratio and lung water content （P<0.01）， and improved pulmonary edema symptoms such as damage， inflammation and infiltration around the lumen， thickening of the trachea， and accumulation of edema fluid， and the SD medium dose group had the best effect on the treatment of PE. In terms of respiratory system， compared with the normal control group， the model group had reduced latent time of cough and asthma （P<0.05， P<0.01） and elevated number of cough and wheezing （P<0.01）. The three SD groups had increased latent time of cough and asthma and decreased number of cough and wheezing （P<0.01）. In terms of urinary system， compared with the normal control group， the model group presented decreased urine volume. The SD low， medium and high dose groups had increased urine volume （P<0.05， P<0.01）， but they had no effect on perspiration. In terms of digestive system， compared with the conditions in the normal control group， the gastric residual rate and gastrin （GT） level were increased （P<0.05， P<0.01）， and the gastric emptying rate and small intestine transit rate were decreased （P<0.01）. The SD low dose group had elevated small intestine transit rate （P<0.01）， and the SD high and medium groups had enhanced gastric emptying rate and small intestine transit rate （P<0.01）， reduced gastric residual rate， lowered GT level to promote gastrointestinal movement and transportation （P<0.01）， and increased motilin （MTL） level to promote gastric emptying in rats （P<0.05， P<0.01）. In terms of circulatory system， compared with the normal control group， the model group displayed reduced left ventricular ejection fraction （LVEF）， left ventricular short axis shortening rate （LVFS） and cardiac output （CO） （P<0.01）， and elevated tendency of heart rate， systolic blood pressure （SBP， P<0.01） and diastolic blood pressure （DBP， P<0.05）. Compared with the model group， the SD low dose group increased LVEF and decreased DBP （P<0.05）， while the SD medium dose group increased LVEF， LVFS， CO and SBP （P<0.01） and decreased DBP （P<0.05）， and the SD high dose group increased LVFS （P<0.01） and decreased SBP （P<0.01） and DBP （P<0.05）. In terms of central nervous system， compared with the conditions in the normal control group， the standing times dropped in the model group （P<0.01）. SD reduced the movement distance， movement time， standing times and activity time in the center of the open field， and increased the rest time and activity time at the edge of the open field （P<0.05， P<0.01）. Moreover， compared with the normal control group， the model group had serious inflammatory infiltration around the lung lumen， thickened trachea with accumulated edema fluid， seriously damaged lung tissue， increased number of white blood cells and percentage of neutrophils in blood （P<0.01）， elevated percentage of monocytes， interleukin-4 （IL-4）， immunoglobulin E （IgE） level and reactive oxygen species （ROS） level in lung tissue （P<0.05，P<0.01）， and decreased IFN-γ in alveolar lavage fluid （P<0.01）. Compared with the model group， SD decreased the number of white blood cells， neutrophil accumulation， pulmonary congestion and interstitial edema， IFN-γ and IL-4 levels in alveolar lavage fluid and ROS level in lung tissue， and increased IgE level （P<0.05， P<0.01）. ConclusionSD had a significant improvement effect on PE model where the disease was located in the viscera. It could regulate the excretion of water by purgation， regulate Qi movement and expel Qi stagnation by descending and sinking lung Qi， and promote purification and descent of lung qi to make Qi movement downward. This indicated SD had the descending and sinking properties. The medium dose of SD decoction exerted the best effect， and its mechanism of action might be through regulating the neutrophil inflammatory response.

Optimization of Extraction Technology of Total Triterpenes from the Leaves of Cornus officinalis by Box-Behnken Design-response Surface Methodology / 中国药房

OBJECTIVE：To optimize the extraction technology for total triterpenes from the leaves of Cornus officinalis . METHODS：Based on the full swelling of the leaves of C. officinalis ，total triterpenes was extracted with heating reflux method. The effects of ethanol concentration ，liquid-solid ratio ，extraction time and extraction times on the contents of total triterpenes from the leaves of C. officinalis were investigated by single factor test. Using oleanolic acid as control ，the contents of total triterpenes were detected by UV spectrometry. On the basis of single factor test ，fixing the times of extraction a s 3 times，taking the contents of total triterpenes as response value ，using ethanol volume fraction ，solid-liquid ratio and extraction time as factors ， Box-Behnken design-response methodology was used to optimize the extraction technology of total triterpenes from the leaves of C. officinalis，and the optimized extraction technology was validated. RESULTS ：The optimal extraction technology of total triterpenes from the leaves of C. officinalis were as follows as ethanol concentration of 73％，liquid-to-material ratio of 38 ∶ 1（mL/g）， extraction time of 60 min. Results of 3 validation tests showed that the contents of total triterpenes from the leaves of C. officinalis were 6.92％，6.91％，6.84％；the average content was 6.89％（RSD＝0.63％），relative error of which with the predicted value （7.28％）was 5.36％. CONCLUSIONS ：The optimized technology is stable and reliable ，and can be used for the extraction of total triterpenes from leaves of C. officinalis .

Exploration on the therapeutic mechanisms and safety of blood components of convalescents without erythrocytes on severe acute respiratory syndrome and coronavirus disease 2019 / 中华危重病急救医学

Coronavirus disease 2019 (COVID-19) is sweeping the world with strong infectivity and high mortality, but by now, we still lack specific treatment. The leading cause of death from COVID-19 is failure to eliminate those viruses effectively. Cellular immunity plays a crucial role in the body's elimination of coronavirus, so it is necessary to study cellular immunity on the basis of plasma therapy. Blood components of convalescents without erythrocytes contains all the components of cellular immunity and humoral immunity in blood. Current studies had shown that all neutrophils, specific antibodies, interferons, platelets, specific effect cells and memory cells, play irreplaceable roles in the immune process of eradicating coronavirus. This article summarizes the infusion safety, therapeutic mechanism of all above components, and their effects on immunologic derangement and excessive inflammatory response, in order to provide an alternative reference for the treatment of COVID-19.

Study on Chemical Constituents of Dioscorea opposita Thunb / 世界科学技术-中医药现代化

This article was aimed to study the chemical constituents of Dioscorea opposita Thunb.The chemical constituents were isolated and purified by Diaion HP-20,Toyopearl HW-40,Sephadex LH-20,MCI Gel CHP-20,silica gel column chromatography and preparative HPLC,TLC,purification and isolation from Dioscorea opposita Thunb.The structures of isolated compounds were identified by thc physicochemical properties and spectral analysis.The result showed that 14 compounds were isolated from Dioscorea opposita Thunb.The chemical structures were elucidated as L-Tryptophane (1),Seguinosides F (2),1-methoxycarbonyl-β-carboline (3),Helichrysin A (4),Bungein A (5),Hydroquinone (6),Zarzissine (7),Cyclo-(Pro-Thr) (8),4-Hydroxy-3-methoxybenzyl alcohol (9),pyridine-3-carboxamide (10),Arbutin (11),Methyl syringate 4-O-β-D-glucopyranoside (12),L-Phenylalanine (13),1,2-benzenedicarboxylic acid,1,2-bis[2-(2-hydroxyethoxy) ethyl] ester (14).It was concluded that chemical compounds 1-14 were isolated for the first time from Dioscorea opposita Thunb.

Effects of five processing methods on compositions and contents of fatty oils in Descurainiae Semen / 中成药

AIM To investigate the effects of frost-like powder,steaming,stir-frying with wine,stir-frying with salt-water and stir-frying with vinegar on compositions and contents of fatty oils in Descurainiae Semen.METHODS Descurainiae Semen was processed by five methods,respectively.The fatty oils were extracted from various processed products by petroleum ether,which were then derivatized.GC-MS was adopted in the qualitative identification and quantitative determination.RESULTS Except for frost-like powder,various processing methods could increase the extraction rate of fatty oils.Compared with raw product,the quantities of fatty oils in various processed products were decreased,together with the increased contents.The main compositions of obtained fatty oils were unsaturated fatty acids,whose contents in various processed products (except stir-frying with vinegar product) were higher than those in the raw product.CONCLUSION The effects of different processing methods on compositions and contents of fatty oils in Descurainiae Semen show obvious differences,among which the processing effect of stir-frying with vinegar is not satisfactory.

Chemical Constituents from Chemical Split Fractions of Seeds ofDescurainia sophia (L.) Webb. ex Prantl / 世界科学技术-中医药现代化

This article was aimed to study the chemical constituents of seeds of Descurainia Sophia (L.) Webb. ex Prantl., in order to lay the material foundation for further interpretation of seeds of D. Sophia and provide pharmacodynamic basis as well as the basis for attributing its nature and taste. The compounds were isolated and purified by Diaion HP-20, Toyopearl HW-40, MCI Gel CHP-20, ODS, Silica gel chromatography combining with Pre-HPLC. The structures were identified on the basis of spectral data and physicochemical properties. Twenty eight compounds were isolated and identified from 20% and 80% ethanol fraction. Thirteen compounds were identified from 20% ethanol fraction: kaempferol-3-O-β-D-glucopyranosyl-7-O-β-D-gentiobioside(1), quercetin-3-O-β-D-glucopyranosyl-7-O-β-D-gentiobioside (2), isorhamnetin-3-O-β-D-glucopyranosyl-7-O-β-D-gentiobioside (3), isorhamnetin-3,7-di-O-β-D-glucopyranoside (4), quercetin-3,7-di-O-β-D-glucopyranoside (5), kaempferol-3, 7-di-O-β-D-glucopyranoside (6), kaempferol-3-O-β-D-xylopyranosyl (1 → 2)-β-D-glucopyranoside (7), methyl sinapate (8), syringaldehyde (9), (S)-p- hydroxyphenyl lactate acid (10), (S)-2-hydroxy-phenylpropionic acid (11), scopoletin (12), sinapic acid (13). Fifteen compounds were identified from 80% ethanol fraction: isorhamnetin-3-O-β-D-glucopyranoside (14), quercetin-3-O-β-D-glucopyranoside (15), kaempferol-3-O-β-D-glucopyranoside (16), quercetin (17), kaempferol (18), isorhamnetin (19), syringic acids (20), quercetin-3-O-β-D-arabinopyranoside (21), quercetin-3-O-β-D-xylopyranoside (22), 6-O-[E]-Sinapoyl-(α- and β)-D-glucopyranoside (23), dimethyl (E, E)-4,4'-dihydroxy-3,3',5,5'-tetramethoxylign-7,7'-dien-9,9'-dioate (24), dimethylthomasidioate (25), 2-hydroxy-3-(1H-indol-3-yl) propanoic acid (26), 2-hydroxyl-3-(1H-indol-3-yl) propanoic acid methyl ester (27), 4'-O-methyl-dihydroquercetin (28). It was concluded that compounds 7-11 and 21-28 were isolated from seeds of D. sophia (L.) Webb. ex Prantl. for the first time.

Inlfuence on Substance and Energy Metabolism of Diabetic Mouse Model of Chemical Split Fractions ofMori Cortex / 世界科学技术-中医药现代化

This article was aimed to study the impact on substance and energy metabolism by chemical split fractions of Mori Cortex among hypoglycemic diabetic mouse model, in order to explain the new hypothesis of the science connotation in nature and flavor of traditional Chinese medicine (TCM). Male Kunming mice were intraperitoneally injected with a large dose of streptozotocin (STZ) (170 mg·kg-1) to establish type 1 diabetes mellitus mouse model. Medication was given consecutively for four weeks. The enzyme-linked immunesorbent assay (ELISA) was used to detect glucosekinase (GCK), glycogen phosphorylase (PYGL), pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (α-KGDHC), phosphoglycerate kinase (PGK), acetyl coenzyme A (acetyl-CoA), adenylate kinase (ADK), fumarase (FUM), cytochrome C reductase (CCR), cytochrome C oxidase (COX) and other indicators. Enzymatic detection was used to determine the content of ATP coenzyme (ATPs), the content and ratio of NAD and NADH, the content of myocardial cell Na+-K+ ATP enzyme, as well as the content of ATP and ADP. The results showed that in the model group, the expression of PYGL was increased; and the expressions of GCK and PDH were decreased. It prompted that the source of glucose increased and the expelling of glucose decreased. The glucose level was increased. The COX expression was reduced and the respiratory chain was blocked. It regulated oxidative phosphorylation and the substrate phosphorylation level. It upregulated the expression of CCR, ATPs, NAD+, PGK, α-KGDHC and ADK. However, the expression of FUM was decreased. The activity of Na+-K+ ATPase was decreased significantly. At last, the metabolic disorders appeared. Mori Cortex aqueous extracts and the chemical split fractions significantly increased the GCK and PDH level in substance metabolism among diabetic mice. The levels of PYGL, α-KGDHC, PGK and acetyl-CoA were decreased (P < 0.05, or P <0.01). Meanwhile, it increased ATP and FUM, myocardial cell Na+-K+ ATP enzyme, and COX level in the energy metabolism (P < 0.05). It decreased the level of NAD+, CCR and ATPs (P < 0.05, or P <0.01). It was concluded that both the aqueous extracts and chemical split fractions of Mori Cortex can effectively improve the substance and energy metabolism disorders of diabetic mouse model. This effect may be related to the cold nature of Mori Cortex.

Study on Chemical Constituents from Chemical Split Fractions ofMori Cortex / 世界科学技术-中医药现代化

This article was aimed to study the chemical constituents from the chemical split fractions of Mori Cortex. The compounds were isolated with Diaion HP-20, Toyopearl HW-40, Sephadex LH-20, MCI Gel CHP-20, Silica gel column chromatography and preparative HPLC. Structures of compounds were identified by physicochemical properties and spectral analysis. The results showed that 23 compounds were obtained. And their structures were identified. The 16 compounds were obtained from the 30% ethanol fraction as vanillic acid (1), 3,4-dimethoxyphenol (2), benzoic acid (3), syringic acid (4), kelampayoside A (5), p-hydroxyphenylpropionic acid (6), caffeic acid (7), hydroferulic acid (8), 6,7-dihydroxycoumarin (9), 5,7-dihydroxycoumarin (10), morin-7-O-β-D-glucopyranoside (11), liriodendrin (12), 2,3-trans-dihydromorin (13), 2,3-cis-dihydromorin (14), 2,3-trans-dihydroquercetin (15), 2,3-cis-dihydroquercetin (16). The 4 compounds were obtained from the 50% ethanol fraction as scopoletin (17), morin (18), kaempferol-7-O-β-D-glucopyranoside (19), umbelliferone (20). The 3 compounds were obtained from the 80% ethanol fraction as sanggenon R (21), cis-mulberroside A (22), resveratrol (23). It was concluded that compounds 2, 4-6, 11, 16, 19 were isolated from this plant for the first time.

Experimental Study on Immunomodulatory Effect of Ethanol Sediments of the seeds of Descurainia sophia(L.) Webb.ex Prantl / 世界科学技术-中医药现代化

This article was aimed to study the immunomodulatory effect of ethanol sediments of the seeds of Descurainia sophia(L.) Webb. ex Prantl. both in vitro and in vivo. The lymphocyte proliferation test in vitro was carried out to explore the effect of the ethanol sediments on the proliferation of T cell and B cell in the spleen of normal mice. And, the carbon clearance test, serum hemolysin test, and delayed-type hypersensitivity test were used to investigate the influence of fraction on non-specific immunity, humoral immunity and cellular immunity in the immunosuppressive mice induced by cyclophosphamide. Besides, the immunosuppressive model was used to evaluate the effect of fraction on immune organs and content of cellular factors in blood serum. The results showed that the ethanol sediments promoted Concanavalin A (Con A) induced T cell and Lipopolysaccharides (LPS) induced B cell (P < 0.01). It increased the carbon clearance index K, phagocytic index α, half value hemolysis (HC50), and swelling degree of auricula (P < 0.05 or P < 0.01). It reduced the body weight and atrophy of thymus and spleen index (P < 0.05 or P < 0.01). It increased the contents of IL-2, IFN-γ and IL-4 in serum in immunosuppressive mice (P < 0.05 or P < 0.01). It was concluded that ethanol sediments of the seeds of D. sophia(L.) Webb. ex Prantl. can boost the lymphocyte proliferation, protect the immune organs, and enhance the non-specific and specific immunity in immunosuppressive mice, which indicated that it had immune-promotion effect.

Research on Fraction-splitting Technology of Seeds ofDescurainia Sophia / 世界科学技术-中医药现代化

This study was aimed to establish the method for fraction-splitting of seeds of Descurainia sophia, in order to study the stability and unoverlapping property of fractions of seeds of D. sophia. The fraction-splitting of seeds of D. sophia were obtained through the combination of various methods including decoction, distillation, extraction, ethanol precipitation and gradient elution of macroporous adsorption resin. HPLC and chemometrics software were used to analyze the stability and unoverlapping property of the fractions of D. sophia. The results showed that the chemical components from seeds of D. sophia was divided into six fractions. HPLC data and chemometrics analysis showed good stability of technology. The fraction-splitting of seeds of D. sophia was unoverlapping. It complied with the research model of chemical constituents of seeds of D. sophia which can be split and combinatorial. It was concluded that the established method for splitting fractions of seeds of D. sophia had good stability and repeatability. Each splitting fraction uncrossed others.

Cloning, Sequence Analysis and Prokaryotic Expression ofGGPS Gene fromLepidium apetalum / 世界科学技术-中医药现代化

This study was aimed to clone the GGPS (geranylgeranyl pyrophosphate synthase) gene from Lepidium apetalum, to analyze its sequence, and to express the protein in E.coli expression system. Specific PCR cloning primers were designed for GGPS gene from Lepidium apetalum according to the full-length sequence from a previous transcriptome sequencing project. PCR amplification was performed with this primer pair on a leaf cDNA template. TA cloning, sequencing and sequence analysis were performed.GGPS gene from Lepidium apetalum was expressed in the E.coli expression system. The results showed that the full-lengthGGPS cDNA from Lepidium apetalum was 1 146 bp coding a protein of 381 amino acids. The LaGGPS protein had an isoprenoid synthase domain. According to a phylogenetic tree constructed with multiple alignment of GGPS protein sequences from various plant species, GGPS protein from Lepidium apetalum was the closest to Arabidopsis thaliana and Sinapis alba. The prokaryotic expression vectorpET-32a-LaGGPS was also constructed successfully. The protein was expressed in E.coli BL21 strain. It was concluded that the cloning and prokaryotic expression of LaGGPS gene provided a foundation for a follow-up research of its function with protein purification and activity analysis.

Study on Essential Oil in the Seeds ofDescurainia Sophia (L.) Webb ex Prantl by GC-MS Analysis / 世界科学技术-中医药现代化

The analysis of essential oil in seeds ofDeseurainia sophia provided an experimental basis for further research on essential oil activity test at the first time. Essential oil was extracted by steam distillation method. Analysis and identification were made by gas chromatography-mass spectrometry (GC-MS) technology in combination with retention indices. A total of 33 components in seeds ofD. sophia were detected by GC-MS and 28 compounds were identified by MS in combination with Kovats retention index. The compounds with high contents were as follows: 3-methylene-nonane (68.14%), 1,3-diazine (29.32%), 2-n-butylacrolein (0.58%), methyl nipecotate (0.43%), 4-oxo-butanenitrile (0.31%), 8-chloro-neoisol-ongifolene (0.25%) and so on. It was concluded that 28 volatile components were identified by GC-MS combined with retention indices. The total detected components were 99.91%. This method was able to improve the accuracy of qualitative detection results.

Experimental Study on Hypoglycemic Effect ofMori Cortex Aqueous Extracts and Chemical Split Fractions / 世界科学技术-中医药现代化

This article was aimed to study effects of aqueous extracts and all chemical split fractions ofMori Cortex on hypoglycemic effect of diabetic mice model. Intra-peritoneal injection of 170 mg·kg-1 streptozocin (STZ) was given to male Kunming mice to establish type I diabetes mode. Continuous administration of medication was given for 4 weeks. And then, indicators such as body weight, water intake, food intake, fasting plasma glucose (FBG), insulin, C-peptide, total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-c), and low density lipoprotein cholesterol (LDL-c) were detected. Pathological morphology of the liver and pancreas were observed by light microscopy. The results showed that high-dose group ofM. Cortex aqueous extracts can improve weight loss of type I diabetic mice, significantly reduce water intake and food intake, reduce FBG, TC, TG and LDL-c levels in different degrees (P<0.05 orP<0.01), and increase C-peptide and HDL-c levels (P<0.05 orP<0.01). When dosages of 30% ethanol fraction and fatty oil fraction were only about 1/2 and 1/4 ofM.Cortex aqueous extracts, we found that it can improve lipid disorder status,repair liver cells, and improve liver tissue damage. Its effect was superior to M. Cortex aqueous extracts. It was concluded thatM. Cortexaqueous extracts showed a better hypoglycemic effect. The effective component parts were 30% ethanol fraction and fatty oil fraction. Its hypoglycemic mechanism may be related to the promotion of insulin secretion, regulation of blood lipid disorders, as well as the protection of liver structure and function.

Study on Diuretic Effect ofMori Cortex Aqueous Extracts and Chemical Split Fractions / 世界科学技术-中医药现代化

This study was aimed to investigate the diuretic effect ofMori Cortex and to identify effective fractions inM. Cortex. Metabolic cages were used to firstly observe the diuretic effect ofM. Cortex aqueous extracts on rats. Medications were continuously given for 5 days to screen for the best dosage of diuretic effect. Picric acid assays were used to detect creatinine levels in serum and urine after 5-day medication. Then, the diuretic effect of each chemical split fraction inM. Cortex was studied in order to indentify the effective parts. The results showed that after administration ofM. Cortex aqueous extracts for 3 to 5 days, compared with the control group, there was a significant diuretic effect on rats (P<0.05 orP<0.01). And the medium-dose ofM. Cortex aqueous extracts showed the best effect (P<0.01). However,M. Cortex aqueous extracts had no significant effect on creatinine levels in serum and urine. Assays of diuretic effect of chemical split fractions ofM.Cortex indicated that compared with the control group, 30% ethanol fraction and fatty oil fraction had the best diuretic effect (P<0.01). It was concluded thatM.Cortex aqueous extracts had a significant diuretic effect. And the chemical fractions contributed mostly to this effect were mainly existed in the 30% ethanol fraction and fatty oil fraction.

Evaluation of Estrogenic Effects ofCoicis Semen Aqueous Extracts / 世界科学技术-中医药现代化

This study was aimed to investigate the estrogenic effects ofCoicis Semen in order to preliminarily discuss the mechanism. Mouse uterine weight test and MCF-7 cell proliferation assay were used to evaluate the estrogenic effects ofC. Semen. Reporter gene assays were adopted to explore the action mechanism ofC. Semen. In reporter gene assay, HEK293 cells were co-transfected with pERE-TAL-luc, pβgal-Control, pCXN2-hERα, or pCXN2-hERβ. And the expression of reporter gene luc was controlled by ERE. Mouse uterine weight test showed that compared with the control group, the aqueous extracts ofC. Semen can increase the uterus index of premature female mice (P<0.01). It can significantly promote the proliferation of MCF-7 cells in the medium without estrogen (P<0.01). The reporter gene controlled by ERE technology showed thatwhen mediated by ERα or ERβ respectively, the normalized luciferase activity of aqueous extracts ofC. Semen was significantly higher than activity of the control group (P< 0.05 orP< 0.01). It was concluded that the aqueous extracts ofC.Semen can increase the uterus index of premature female mice and promote the proliferation of MCF-7 cells in the medium without estrogen. We found the estrogenic effects ofC. Semen for the first time. And the estrogenic effects ofC. Semen were mainly mediated by ERβ.

Protection effect of amentoflavone in Selaginella tamariscina against TNF-alpha-induced vascular injury of endothelial cells / 药学学报

This study is to observe the protection effect of amentoflavone (AMT) in Selaginella tamariscina against TNF-alpha-induced vascular inflammation injury of endothelial cells. On the basis of TNF-alpha induced human umbilical vein endothelial cell, observe the influence of AMT on endothelial active factor, the contents of SOD and MDA, the protein expression of vascular endothelial adhesion molecules and inflammatory factor; study the effect of its common related signal pathways such as NF-kappaB; research the effect of AMT against TNF-a induced human umbilical vein endothelial cell injury by means of MTT, ELISA, Western blotting and the cell immunofluorescence. The results showed that AMT could increase the content of NO and decrease the levels of VCAM-1, E-selectin, IL-6, IL-8 and ET-1; enhance the activity of SOD, reduce the content of MDA; downregulate the protein expressions of VCAM-1, E-selectin, NF-kappaBp65 and up-regulate IkappaBalpha, attenuate the NF-kappaBp65 transfer to cell nucleus. AMT has the effect of protect vascular endothelial and maybe via the signal pathway of NF-kappaB to down-regulate the inflammation factor and oxidative damage factor of downstream.