Mechanism of Flavonoids from Mulberry Leaves on Insulin Resistance and Inflammatory Response in Diabetic Mice / 中国实验方剂学杂志

ObjectiveTo observe the glucose-lowering， insulin resistance-improving， and anti-inflammatory effects of flavonoids from mulberry leaves （FML） and explore their underlying mechanism. MethodMale db/db mice aged 6-7 weeks were randomly divided into a model group， a high-dose FML group （1.00 g·kg·d-1）， and a low-dose FML group （0.50 g·kg-1·d-1）. C57BL mice of the same age were assigned to the normal group. After six weeks of intervention， fasting blood glucose （FBG）， serum fasting insulin levels （Fins）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， free fatty acid （FFA）， blood creatinine （SCr）， blood urea nitrogen （BUN）， and aspartate aminotransferase （AST） levels were measured， and the homeostasis model assessment of insulin resistance （HOMA-IR） was calculated. Superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， and catalase activities in the liver were measured. Morphological changes in the liver were assessed by hematoxylin-eosin （HE） staining. The protein expression of cyclooxygenase-2 （COX-2）， inducible nitric oxide synthase （iNOS）， and nuclear factor-κB （NF-κB） in the liver was detected by Western blot. ResultCompared with the model group， the high-dose and low-dose FML groups showed significant reductions in FBG， Fins， HOMA-IR， IL-6， TNF-α， and FFA levels （P<0.05， P<0.01）， and increased levels of SOD， GSH-Px， and catalase in the liver （P<0.05， P<0.01）. HE staining of the liver in the FML groups showed improved arrangement of hepatocytes， reduced inflammatory cell infiltration， and alleviated cellular steatosis compared with the model group. The protein expression of COX-2， iNOS， and NF-κB in the liver significantly decreased in the FML groups as compared with that in the model group （P<0.05， P<0.01）. ConclusionFML have glucose-lowering and insulin resistance-improving effect， which may be attributed to their regulation of the NF-κB pathway in the liver of diabetic mice， leading to the suppression of the release of COX-2， iNOS， and inflammatory cytokines， thereby improving the inflammatory state.

Nature-effect transformation mechanism of mulberry leaves and silkworm droppings based on chemical composition analysis / 中国中药杂志

Starting with the relationship between mulberry leaves and silkworm droppings as food and metabolites, this study systematically compared the chemical components, screened out differential components, and quantitatively analyzed the main differential components based on ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and UPLC-Q-TRAP-MS combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Moreover, the in vitro enzymatic transformation of the representative differential components was studied. The results showed that(1) 95 components were identified from mulberry leaves and silkworm droppings, among which 27 components only exist in mulberry leaves and 8 components in silkworm droppings. The main differential components were flavonoid glycosides and chlorogenic acids.(2) Nineteen components with significant difference were quantitatively analyzed, and the components with significant differences and high content were neochlorogenic acid, chlorogenic acid, and rutin.(3) The crude protease in the mid-gut of silkworm significantly metabolized neochlorogenic acid and chlorogenic acid, which may be an important reason for the efficacy change in mulberry leaves and silkworm droppings. This study lays a scientific foundation for the development, utilization, and quality control of mulberry leaves and silkworm droppings. It provides references for clarifying the possible material basis and mechanism of the pungent-cool and dispersing nature of mulberry leaves transforming into the pungent-warm and dampness-resolving nature of silkworm droppings, and offers a new idea for the study of nature-effect transformation mechanism of traditional Chinese medicine.

Efficacy and Safety of Mulberry Twig Alkaloids Tablet for Treatment of Type 2 Diabetes: A Randomized, Double-Blind, Placebo-Controlled Multicenter Clinical Study / 中国结合医学杂志

OBJECTIVE@#To assess the efficacy and safety of mulberry twig alkaloids (Sangzhi alkaloids, SZ-A) for treatment of type 2 diabetes in a randomized, double-blind, placebo-controlled multicenter clinical trial.@*METHODS@#A total of 200 patients were randomized to receive SZ-A (n=100) or placebo (n=100) for 16 weeks. The data analysis system for electronic data capture clinical trial central randomization system was used for randomization and dispensing of drugs. The primary outcome was the change in glycosylated hemoglobin (HbA1c) level. The secondary outcome included the proportions of cases with HbA1c <7.0% and HbA1c <6.5%, fasting blood glucose (FBG), postprandial blood glucose (PBG), area under curve for the PBG (AUC0-2h), body weight, and body mass index (BMI). Adverse events (AEs), severe adverse events (SAEs), treatment-related adverse events (TAEs), gastrointestinal disorders (GDs), blood pressure, routine blood tests, and liver and kidney function were monitored.@*RESULTS@#Compared with baseline, the change of HbA1c at week 16 was -0.80% (95% CI: -0.98% to -0.62%) and -0.09% (95% CI: -0.27% to 0.09%) in SZ-A group and placebo group, respectively. The proportion of patients with HbA1c <7% and <6.5% was higher in the SZ-A group than in the placebo group (46.8% vs. 21.6% and 29.9% vs. 10.8%). The observed values and changes in FBG, 1 h-PBG, 2 h-PBG, and AUC0-2h differed significantly between groups (P<0.001), but differences were not significant in body weight and BMI (P>0.05). The incidence rates of AEs, TAEs, and GDs differed significantly between groups (P=0.010, P=0.005, and P=0.006, respectively), whereas the incidence rates of SAEs showed no significant differences between groups (P=1.000).@*CONCLUSION@#SZ-A are effective and safe for treatment of type 2 diabetes. The protocol was registered in http://www.chictr.org.cn/showproj.aspx?proj=60117 (ChiCTR2000038550).

A Preliminary Study for the Suppressive Effect of Mulberry Leaf Powder-containing Foods on the Postprandial Blood Glucose Level / 日本補完代替医療学会誌

A crossover study was conducted to evaluate suppressive effect of a commercially available green juice (Katuna-Aojiru;Egao Co., Ltd.) containing mulberry leaf powder as the main ingredient on postprandial hyperglycemia. The blood glucose and iAUC (0-120min) values after loading cooked white rice were significantly lower with the consumption of Katuna-Aojiru than with the consumption of water. Katuna-Aojiru is effective in controlling postprandial blood glucose.

Total Flavonoids of Mulberry Leaves Improves Liver Lipid Metabolism in Type 2 Diabetic Rats by Regulating PPAR-α/CPT-1 Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the medicinal effect of total flavonoids of mulberry leaves on regulating liver lipid metabolism disorder in diabetes mellitus type 2 （T2DM） rats， and the mechanism based on liver peroxidase proliferators activate receptors-α （PPAR-α） and carnitine palmityl transferase-1 （CPT-1） proteins. MethodTotal flavonoids of mulberry leaves were extracted and purified by ethanol extraction + macroporous resin purification and then identified. T2DM rat model was induced by high fat diet （HFD） + streptozocin（STZ）method. Rats with blood glucose ≥ 11.1 mmol·L-1 were divided into three administration groups with the high dose （300 mg·kg-1）， medium dose （150 mg·kg-1）， and low dose （75 mg·kg-1） of total flavonoids of mulberry leaves for 8 weeks， respectively， to observe the weight and blood glucose of the rats. The pathological changes of rat livers were observed by hematoxylin-eosin （HE） staining. Biochemical method was used to detect the levels of total cholesterol （TC）， triglyceride （TG）， low density lipoprotein-cholesterol （LDL-C）， and high density lipoprotein-cholesterol （HDL-C） of blood lipid metabolism in rats. The messenger ribonucleic acid （mRNA） and protein expressions of PPAR-α and CPT-1 were determined by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultAfter 8 weeks of intervention of total flavonoids of mulberry leaves， compared with the control group， the food intake， liver index， and fasting blood glucose of rats in the model group increased significantly （P<0.01）. Compared with the model group， the food intake， fasting blood glucose， and liver index of rats in the administration groups decreased significantly （P<0.01）. The results of HE staining showed that the liver tissue structure of rats in the control group was complete and there was no obvious abnormality. The model group showed vacuolar degeneration and inflammatory infiltration of hepatocytes of rats. There was no obvious abnormality in the liver structure of rats in the administration groups. The results of blood lipid showed that compared with the control group， the levels of TC， TG， and LDL-C increased significantly （P<0.01）， but the level of HDL-C decreased significantly （P<0.01） in the model group. Compared with the model group， the levels of TC， TG， and LDL-C decreased significantly （P<0.05， P<0.01）， whereas the level of HDL-C increased significantly （P<0.01） in the administration groups. The results of Real-time PCR showed that compared with the control group， the mRNA expression of PPAR-α and CPT-1 of rats in the model group decreased significantly （P<0.01）. Compared with the model group， the mRNA expressions of PPAR-α and CPT-1 of rats in the high-dose group increased significantly （P<0.01）. The results of Western blot showed that compared with the control group， the protein expressions of PPAR-α and CPT-1 of rats in the model group decreased significantly （P<0.01）. Compared with the model group， the protein expressions of PPAR-α and CPT-1 of rats in the high-dose group increased significantly （P<0.05， P<0.01）. ConclusionTotal flavonoids of mulberry leaves can effectively reduce blood glucose and improve liver lipid metabolism disorder in T2DM rats. The total flavonoids of mulberry leaves could regulate lipid metabolism and play a hypoglycemic role by activating and regulating PPAR-α and CPT-1 proteins and promoting oxidative decomposition of fatty acids.

Discussion on Material Basis of Different Parts of Mulberry with Different Effects Based on Molecular Connectivity Index / 中国实验方剂学杂志

ObjectiveTo explore the material basis for the difference in the efficacy of different parts of mulberry based on molecular connectivity index （MCI）. MethodBy referring to the relevant literature at home and abroad and traditional Chinese medicine systems pharmacology database and analysis platform （TCMSP） database， the chemical composition database of mulberry-source medicinal materials was established. Venn analysis was carried out on the components among mulberry-source medicinal materials. The components in the database were divided into 10 categories， and the composition information was analyzed. According to MCI value， all components of mulberry-source medicinal materials were divided into different groups. The angle cosine method was used to calculate the MCI similarity. The average MCI values of the common component group from 0-8 orders and CI of mulberry-source medicinal materials were calculated. ResultThe components with high similarity such as （+）-cycloolivil， 1′-methoxy-2′-hydroxydihydromollugin， kuwanon， morusin and 1-deoxynojirimycin were selected as potential pharmacodynamic components. Mulberry-source medicinal materials could be divided into five component groups. The similarity between component groups and total components was 0.760-0.999， and the similarity between component groups was 0.248-0.999. In Mori Ramulus， Mori Folium， Mori Cortex and Mori Fructus， the average MCI values of their flavonoids from 0-8 orders were 4.57， 4.59， 6.41， 4.24， respectively. The average MCI values of alkaloids from 0-8 orders were 2.65， 4.55， 2.58， 2.78， respectively. The average CI values from 0-8 orders were 5.51， 5.49， 5.44 and 2.88， respectively. ConclusionIt is preliminarily concluded that there are differences in the flavonoids and pathways of hypoglycemic effects between Mori Cortex and the other three mulberry-source medicinal materials. The MCI values of alkaloids from 0-8 orders in Mori Folium and Mori Fructus were higher， but their inhibitory activity of α-glucosidase were lower than those of Mori Ramulus and Mori Cortex. The structural characteristics of the total components of Mori Fructus represented by CI were quite different from the other three mulberry-source medicinal materials.

Research progress in mulberry leaf polysaccharide in treating diabetic nephropathy / 中国药理学与毒理学杂志

Diabetic nephropathy (DN) is one of the most common complications of diabetes. It is an important cause of diabetes disability and death. DN is a systemic metabolic syndrome. In its pathogenesis, the interaction of various cell activities and a large number of cytokine biological activities, the activation of signal pathways and so on are involved in the development of DN. At present, the clinical treatment of DN is mainly Western medicine, but it has limitations such as strong toxicity, high side effects and poor compliance. Therefore, the discovery of natural anti-DN substances has also become an important means to treat DN. Mulberry leaves are the dry leaves of Morus alba L. It is not only a tradi?tional Chinese medicine, but also a dual-purpose medicinal material for medicine and food. It has the effects of dispelling wind and clearing heat, cooling blood and brightening eyes, tonifying and so on. Mulberry leaf polysaccharide (MLP) is a kind of high molecular compound in mulberry leaves. It has many pharmacological effects, such as hypoglycemic, antiox?idant, anti-stress, anti-virus and so on. Therefore, the pharmacological effects of mulberry leaf polysaccharides on dia?betic nephropathy are reviewed in this paper, so as to provide references for further research and application. The patho?genesis of DN is complex, and the mechanism of renal injury has not been completely clarified. The current studies believe that DN is closely related to heredity, abnormal glucose metabolism, abnormal lipid metabolism, microcirculation disorder, cytokine action, oxidative stress and so on. Relevant studies show that the pharmacological effects of mulberry leaf polysaccharide in the prevention and treatment of DN mainly include: ① Effect on transforming factor-β1 (TGF-β1):TGF-β1 has become an important cytokine involved in the formation of renal fibrosis by regulating cell proliferation and differentiation and the production of extracellular matrix (ECM). MLP can significantly inhibit TGF-β1 protein, and then inhibit the synthesis of extracellular matrix by renal interstitial fibroblasts and inhibit the realization of fibrosis.②Effect on insulin receptor substrate (IRS-1): IRS-1 is an important signal molecule at the beginning of IR signal transduction. The decrease of IRS-1 gene expression or the decrease of expression can affect the effective transmission of IR signal and lead to the development and deterioration of diabetes. MPL can significantly increase the expression of IRS-1 mRNA in liver tissue of DN rats, so as to prevent and treat DN. ③ Effect on the expression of resistin protein in adipose tis?sue. Resistin is a secretory polypeptide derived from adipose tissue and is specifically expressed in white adipose tissue and is closely related to type 2 diabetes mellitus (T2DM). Experimental studies show that MLP can effectively reduce the expression of resistin protein in white adipose tissue of T2DM rats, indicating that MLP may reduce the level of IR by inhibiting the expression of resistin in adipose tissue, thereby reducing the insulin resistance state of T2DM rats, so as to achieve the goal of treating diabetes.④Effect on adiponectin receptor 1 (AdipoR1):adiponectin can improve insulin resistance, reduce blood glucose and lipid. AdipoR1 is mainly expressed in skeletal muscle and kidney. Studies have shown that AdipoR1 is closely related to the occurrence and development of DN. The results showed that MLP could reduce the blood glucose and blood lipid level and up regulate the expression of AdipoR1 mRNA in DN rats, suggesting that MLP may delay the occurrence and development of DN. This article reviewed the pharmacological effects of mulberry leaf polysaccharides on diabetic nephropathy, and provided a useful basis for further development and utilization of mul?berry leaf polysaccharides in the treatment of DN.

Combined enzymatic hydrolysis and herbal extracts fortification to boost in vitro antioxidant activity of edible bird's nest solution / 中草药·英文版

Objective: Edible bird's nest (EBN) is a popular traditional tonic food in Chinese population for centuries. Malaysia is one of the main EBN suppliers in the world. This study aims to explore the best strategy to boost the antioxidant potential of EBN solution. Methods: In this study, the raw EBN (4%, mass to volume ratio) was initially enzymatic hydrolyzed using papain enzyme to produce EBN hydrolysate (EBNH), then spray-dried into powdered form. Next, 4% (mass to volume ratio) of EBNH powder was dissolved in ginger extract (GE), mulberry leaf extract (MLE) and cinnamon twig extract (CTE) to detect the changes of antioxidant activities, respectively. Results: Results obtained suggest that enzymatic hydrolysis significantly reduced the viscosity of 4% EBN solution from (68.12 ± 0.69) mPa·s to (7.84 ± 0.31) mPa·s. Besides, the total phenolic content (TPC), total flavonoid content (TFC), total soluble protein, DPPH scavenging activity and ferric reducing antioxidant power (FRAP) were substantially increased following EBN hydrolysis using papain enzyme. In addition, fortification with GE, MLE and CTE had further improved the TPC, TFC, DPPH scavenging activity and FRAP of the EBNH solution. Among the samples, MLE-EBNH solution showed the most superior antioxidant potential at (86.39 ± 1.66)% of DPPH scavenging activity and (19.79 ± 2.96) mmol/L FeSO

Submerged fermentation improves bioactivity of mulberry fruits and leaves / 中草药·英文版

Objective: Mulberry (Morus spp.) fruits and leaves have been proven to possess nutraceutical properties. Due to its fast and easy growing characteristics, mulberry fruits (MF) and leaves (ML) potentially emerge as a great source of functional foods. This study aims to enhance bioactivities (antioxidant, anti-inflammation, and hypoglycemic activity) of MF and ML via submerged fermentation using bacteria (Lactobacillus plantarum TAR 4), yeast (Baker's yeast and red yeast) and fungi (Tempeh and Tapai starter). Methods: In this study, 25% (mass to volume ratio) of MF and ML were fermented (48 h) with 1% (mass to volume ratio) of different microbial cultures, respectively. Effects of different fermentations on MF and ML were determined based on the changes of total phenolics (TPC), flavonoids (TFC), anthocyanins, total sugar, DPPH activity, ferric reducing antioxidant power (FRAP), albumin denaturation inhibition activity (ADI), anti-lipoxygenase activity and α-amylase inhibition activity (AI). Results: Generally, ML had higher AI than MF. However, MF exhibited higher DPPH, FRAP and anti-lipoxygenase activity than ML. After all forms of fermentation, DPPH and AI activity of MF and ML were increased significantly (P < 0.05). However, the effects of fermentation on TPC, FRAP, ADI and anti-lipoxygenase activity of MF were in contrast with ML. TPC, FRAP and anti-lipoxygenase activity of ML were enhanced, but reduced in MF after fermentation. Although the effects exerted by different microorganisms in MF and ML fermentation were different, the bioactivities of MF and ML were generally improved after fermentation. Fermentation by Tempeh starter enhanced TPC (by 2-fold), FRAP (by 2.3-fold), AI (at 10% increment) and anti-lipoxygenase activity (by 5-fold) of ML, whereas Tapai fermentation effectively enhanced the DPPH (at 17% increment) and ADI (by 2-fold) activity of MF. Conclusion: Findings of this study provide an insight into the future process design of MF and ML processing into novel functional foods.

Comparative analysis and research on composition of mulberry-sourced medicinal materials by fingerprint division total statistical moment method and information entropy / 中国中药杂志

The present work is to analyze the HPLC fingerprints of mulberry-sourced materials(Mori Ramulus, Mori Folium, Muri Cortex, Mori Fructus) using the fingerprint division total statistical moment method and information entropy, and to study the diffe-rences of the chemical components and the overall characteristics of the imprinting template in different parts of mulberry-sourced medicinal materials, so as to provide the basis for finding the effective substances in response to "homologous and different effect" of mulberry(Morus alba). The fingerprints of 24 batches of mulberry-related materials, such as Mori Ramulus, Mori Folium, Muri Cortex, Mori Fructus, were established, and the similarities and differences of the fingerprints were analyzed by calculating the division total statistical moment parameters and information entropy. The AUC_T, MCRT_T, VCRT_T and H values of 24 batches of mulberry-sourced materials were less than 0.05, with significant difference. Among them, all samples showed absorption peaks within 3-11, and 20-24 min, indicating that the four types had the identical or similar chemical composition in the same time period. After 34 min, none of the four types showed absorption peaks. Greater VCRT_T value of the fingerprints of the four kinds was observed at the retention time ranges of 3-4, 16-18, 25-27, and 31-32 min, indicating that the components of the four kinds were significantly different in these time periods; and VCRT_T value of the mulberry was significantly higher than that of the other three kinds of medicinal materials at the retention time windows of 3-4 and 15-17 min; the VCRT_T value of the mulberry white skin was significantly higher at the time windows of 8-10 and 28-30 min; the VCRT_T value of all four kinds was significantly higher within 21-23 min, indicating that the four herbs contain the same or similar components in the chromatogram during this period, but there may be significant differences between the content and the proportion. In addition, the information entropy of mulberry branches is the largest at 7-12, 23-27 min, and that of mulberry fruits is the largest at 2-8 min, which indicates that the components of mulberry branches and mulberry fruits respond greatly in the corresponding period of time, which is also the main peak period of their chemical components. For the chemical components and corresponding efficacy here. The results showed that there are significant differences in the components and contents of mulberry-sourced medicinal materials. The division total statistical moment and information entropy of the total amount of segments can be used to analyze the differences in the components of "homology and different effects", which could provide a more comprehensive analysis method for the determination of quality markers.

Mulberry polysaccharides improves the chemotherapy of liver cancer ascites tumor-bearing mice by regulating the PI3K/AKT/mTOR pathway / 中国临床药理学与治疗学

AIM: To study the synergistic and attenuating effects of mulberry polysaccharides on the chemotherapy of liver cancer ascites tumor-bearing mice by regulating the PI3K/AKT/mTOR pathway. METHODS: Ninety SPF male Kunming mice were randomly divided into normal group, model group, cyclophosphamide group, mulberry polysaccharide group and mulberry polysaccharide + cyclophosphamide group. Liver cancer ascites tumor-bearing mice were prepared, and each administration group was given 30 mg/kg of cyclophosphamide or (and) 200 mg/kg of mulberry polysaccharide and administered orally. The normal group and the model group were administrated with 10 mL/kg saline. The tumor suppression rate, liver, spleen and other indexes, tumor tissue VEGF and inflammatory factor content, and PI3K/AKT/mTOR pathway-related protein expression were observed in mice.RESULTS:Cyclophosphamide group, mulberry polysaccharide group and mulberry polysaccharide + cyclophosphamide group mice body weight, tumor mass, tumor tissue VEGF, TNF-α, IL-6, PI3K, AKT and mTOR phosphorylation levels were lower than the model group, and the level of IL-1β was higher than the model group; mulberry polysaccharide + cyclophosphamide group mice were observed with body weight, tumor inhibition rate, tumor tissue VEGF, TNF-α, IL-6, PI3K, AKT and The mTOR phosphorylation level higher than the mulberry polysaccharide group and cyclophosphamide group, and the tumor mass and IL-1β level were lower than the mulberry polysaccharide group and cyclophosphamide group (P<0.05). CONCLUSION: Morus alba polysaccharide combined with cyclophosphamide can effectively inhibit tumor proliferation of liver cancer ascites tumor-bearing mice and exert attenuating effect. The mechanism may be related to the down-regulation of PI3K /AKT/mTOR pathway expression.

Fermentation of mulberry leaves with Cordyceps militaris enhanced anti-adipogenesis activity in 3T3-L1 cells through down-regulation of PPAR-? pathway signaling

Objective: To establish an efficacious and efficient fermentation method of enhancing the anti-adipogenesis effect of mulberry (Morus alba) leaves using Cordyceps militais. Methods: Dried mulberry leaves, dried mulberry leaves with 50% raw silkworm pupa and raw silkworm pupa were fermented with Cordyceps militais for 4 weeks at 25 °C, after which the dried mulberry leaves and fermented product were extracted with 70% ethanol and subjected to high performance liquid chromatography (HPLC). The contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid were determined. We then used the 3T3-L1 cells to investigate whether extracts of fermentation enhanced anti-adipogenesis activity in vitro. Results: HPLC showed that fermentation changed the contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid. Furthermore, fermented dried mulberry leaves with 50% raw silkworm pupa had a better efficacy of anti-adipogenesis than dried mulberry leaves, fermented dried mulberry leaves and fermented silkworm pupa and inhibited triglycerides accumulation and glucose consumption. Additionally, fermented dried mulberry leaves with 50% raw silkworm pupa inhibited PPAR-? signaling. Conclusions: Fermentation with Cordyceps militaris enhanced anti-adipogenesis efficacy of mulberry leaves.

The application of Mulberry eye drops on xerophthalmia in perimenopausal female rabbit model / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the therapeutic efficiency of Mulberry eye drops on dry eye in perimenopausal female rabbit model. <p>METHODS: Totally 36 female New Zealand white rabbits were selected and operated with bilateral ovariectomy and fed for 60d postoperatively to make perimenopausal female rabbit dry eye model(36 eyes, all right eyes). And 24 of them were randomly divided into control group and experimental group(12 for each): the control group used PBS, the experimental group used Mulberry eye drops, the other 12 did not use any eye drops. The Schirmer I tests(SⅠt)and corneal fluorescein(FL)were made, and the tear total protein content, amylase activity, lactoferrin and lysozyme contents and confocal scanning microscopy were performed in two groups before treatment and 2, 4, 6wk after treatment. <p>RESULTS: There was no significant difference in SⅠt,FL scores,total proteins,lysozyme,lactoferrin contents and amylase activity between two groups at pre-therapy(all <i>P</i>>0.05).After 2, 4 and 6wk of treatment, there were significant changes in SⅠt and FL scores in the two groups(<i>P</i><0.05). There were no significant changes in lysozyme content, lactoferrin, amylase activity and total tear protein in the experimental group compared with those before treatment(all <i>P</i>>0.05), while there were significant changes in lysozyme content, lactoferrin, amylase activity and total tear protein in the control group(all <i>P</i><0.05). At 2, 4 and 6wk after treatment, there were differences in the scores of SⅠt and FL, lysozyme content, lactoferrin, amylase activity and total protein content in tear between the two groups(<i>P</i><0.05). At 6 wk after treatment, the mean number of corneal basal cells and inflammatory cells in control group were 4436±289mm2 and 321±91mm2,respectively,which in experimental group were 3219±223mm2 and 36±11mm2, respectively,there were statistical differences between two groups( all <i>P</i><0.05). After 6wk treatment, there were no change of corneal nerve bending and less density in control group while the nerve fiber bending and density decreased significantly in experimental group,there were statistical differences between two groups(all <i>P</i><0.05).<p>CONCLUSION: Mulberry eye drops has significantly therapy effects on the dry eye induced by decreased androgen and it has a definite clinical application value.

Study on Protection Effect of Mulberry Anthocyanin- 3-glucoside on Epilepsy Model Mice and the Effect of Hippocampal BDNF/TrkB Pathway / 中国药房

OBJECTIVE：To study the protection ef fects of mulberry anthocyanin- 3-glucoside on epilepsy model mice and the effect of hippocampal brain derived neurotrophic factor （BDNF）/tyrosine kinase B （TrkB）pathway. METHODS ：Totally 120 C57BL/6 mice were randomly divided into normal group ，model group ，single medication group （mulberry anthocyanin- 3- glucoside），agonist combination group（mulberry anthocyanin- 3-glucoside+TrkB agonist LM 22B-10），with 30 mice in each group. single medication group and agonist combination group were given mulberry anthocyanin- 3-glucoside 600 μg/kg intragastrically once a day ，for consecutive 6 weeks. The agonist combination group was given LM22B-10（5 mg/kg）via the lateral ventricle once a day at 6th week. Normal group and model group were given constant volume of normal saline intragastrically. After last medication，except for normal group ，other groups were given lithium chloride-pilocarpine to establish epilepsy model. After modeling，10 mice in each group were taken to record the latency ，frequency and duration of spontaneous recurrent epilepsy ， observed for 6 hours a day for 4 weeks；EEG was recorded on the 14th，28th and 36th day after modeling ，and the abnormal frequency of EEG in 1 h was counted . On the 6th day of modeling ，other 10 mice in each group were taken to detect the serum calcium level ，and the remaining 10 mice in each group were taken to detect the expressions of BDNF mRNA and protein in the hippocampus. RESULTS ：Compared with normal group ，latency，frequency and duration of spontaneous recurrent epilepsy and the times of abnormal brain wave on the 14th，28th and 36th day after modeling were increased significantly in model group （P＜ 0.05）. The serum calcium level ， mRNA and proteinexpression of BDNF in hippocampus were increased E-mail：wangfang7699@126.com significantly （P＜0.05）. Compared with model group ，the latency，frequency，duration of spontaneous recurrent epilepsy and the times of abnormal brain wave on the 28th and 36th day after modeling were decreased significantly in single medication group（P＜0.05），while serum calcium level ，mRNA and protein expression of BDNF in hippocampus were decreased significantly （P＜0.05）. Compared with single medication group ，the latency，frequency and duration of spontaneous recurrent epilepsy and the times of abnormal brain wave on the 28th and 36th day after modeling were increased significantly in agonist combination group （P＜0.05），while serum calcium level ，mRNA and protein expressions of BDNF in hippocampus were increased significantly （P＜0.05）. CONCLUSIONS ：Mulberry anthocyanin- 3- glucoside has a good protection effect on epilepsy model mice ，the mechanism of which may be associated with inhibiting the activation of hippocampal BDNF/TrkB pathway.

Fermentation of mulberry leaves with Cordyceps militaris enhanced anti-adipogenesis activity in 3T3-L1 cells through down-regulation of PPAR-? pathway signaling

Objective: To establish an efficacious and efficient fermentation method of enhancing the anti-adipogenesis effect of mulberry (Morus alba) leaves using Cordyceps militais. Methods: Dried mulberry leaves, dried mulberry leaves with 50% raw silkworm pupa and raw silkworm pupa were fermented with Cordyceps militais for 4 weeks at 25 °C, after which the dried mulberry leaves and fermented product were extracted with 70% ethanol and subjected to high performance liquid chromatography (HPLC). The contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid were determined. We then used the 3T3-L1 cells to investigate whether extracts of fermentation enhanced anti-adipogenesis activity in vitro. Results: HPLC showed that fermentation changed the contents of cordycepin, pelargonidin, chlorogenic acid, iso-quercetin and caffeic acid. Furthermore, fermented dried mulberry leaves with 50% raw silkworm pupa had a better efficacy of anti-adipogenesis than dried mulberry leaves, fermented dried mulberry leaves and fermented silkworm pupa and inhibited triglycerides accumulation and glucose consumption. Additionally, fermented dried mulberry leaves with 50% raw silkworm pupa inhibited PPAR-? signaling. Conclusions: Fermentation with Cordyceps militaris enhanced anti-adipogenesis efficacy of mulberry leaves.

Genetic characterization of silkworm (Bombyx mori L. ) strains (Lepidoptera: Bombicidae) with different geographical originon the basis of isozyme markers

The aim of the present study was to assess genetics variations within and among strains with different geographical origin, belonging to Bulgarian germplasm bank of mulberry silkworm (Bombyx moriL.)and to establish their relationships using isoenzyme markers. Polyacrilamide gel electrophoresis (PAGE) was used to study the isoenzymes of nonspecific esterases (EST), malate dehydrogenase (MDH) and acid phosphatase (ACP) from haemolymph, phosphoglucomutase (PGM) and hexokinase (HK) from silk glands and alkaline phosphatase (ALP) from midgut of mulberry silkworm (Bombyx moriL.). Variability was evident in all of these enzyme systems among twelve strains from Bulgarian germplasm resources of B. mori.Total of nine lociwere detected. All of them (100%) were polymorphic. “Null” alleles in four lociwere determined. Intra-and inter-strain polymorphism was obtained. The degree of polymorphism ranged from 0% to 77.80%. Low levels of observed heterozygosity in comparison with the expected one have been calculated in almost all of strainsas well as deviations from Hardy-Weinberg equilibrium in almost all analyzed loci, resultingof excess of homozygotes. The value of FSTwas 0.4903. The dendrogram constructed with the values of genetic distance manifests that Romanian strain Cislau Tokay is formed one main clade while the rest strains studied (from Bulgaria, Japan, China, Vietnam, Spain, Syria and Egypt) are formed the other clade withsubclades. The genetic data of the tested strains from different geographical regions, would be used for identifying suitable parents for breeding programs with a view to yield improvement in this species with economic significance.

Effects of the mulberry leaf extract on depression-like behavior in mice / 国际药学研究杂志

Objective: To investigate the antidepressant effect and mechanism of the extract of mulberry leaf(EML). Methods: Male BALB/c mice were randomly assigned to receive EML(100, 200 and 400 mg/kg)or vehicle for 7 days or single administration by intragastric(ig)route. The effect of EML on spontaneous activity in mice was evaluated using the open field test. The anti-depressant activity of EML was evaluated using the tail suspension test and forced swimming test. After single ig administration of EML (100, 200 and 400 mg/kg), the effects of EML on the function central monoaminergic nervous system were evaluated using the 5-hydroxy-L-tryptophan(5-HTP)induced head-twitch test, yohimbine toxicity potentiation test and reserpine test in mice. Results: In the behavioral despair model, the continuous administration of EML(100 mg/kg)for 7 days significantly reduced the immobility time in the tail suspension test and forced swimming test. Compared with the vehicle group, the inhibitory rates of immobility time were 35.5% and 41%(P<0.05, P<0.01)in the tail suspension test and forced swimming test, respectively. The single treatment with EML(400 mg/kg)also significantly reduced the immobility time in the tail suspension and forced swimming test, and the inhibitory rates of immobility time were 29.1% and 35.3%, respectively(P<0.05). The results of spontaneous activity test showed that EML had no excitatory or inhibitory effect on the central nervous system in mice. In the 5-HTP induced head-twitch test in mice, the single treatment with EML(100 mg/kg)significantly increased the number of head-twitches in mice. In the yohimbine toxicity potentiation test, the single treatment with EML(100, 200 and 400 mg/kg)had no significant effect on the mortality rate in yohimbin-treated mice. In the reserpine test, compared with the model group, the single treatment with EML(400 mg/kg)antagonized reserpine induced ptosis (P<0.05)and had no significant effect on the decrease in rectal temperature and akinesia. Conclusion: EML showed antidepressant effect, and the action mechanism was likely related to the enhancement of the function of serotonergic nervous system.

Role of growth regulators and plant water extracts on the productivity and water use efficiency of wheat genotypes under limited water supply / Papel dos reguladores de crescimento e dos extratos de plantas sobre a eficiência da produtividade e do uso da água dos genótipos de trigo sob abastecimento de água limitada

Empowerment of wheat genotypes by application of growth regulators, compatible solutes and plant extracts under water restriction is an important strategy for getting sustainable yield. This study aimed to evaluate the effects of drought stress on the growth and yield of wheat genotypes and also monitor and compare the role of ABA, SA as well as moringa and mulbery leaf water extracts in improving drought tolerance of wheat genotypes. The work was performed at the research area of the Faculty of Agricultural Sciences, Ghazi University, Dera Ghazi Khan, Pakistan. Three wheat cultivars Aas-2011, Faisalabad- 2008 and Triple dwarf-1 were subjected to drought stress (skipping the irrigation at grain filling stage). The wheat genotypes were subjected to treatments viz., T1 i.e. All normal irrigation without application of abscisic acid (ABA), salicylic acid (SA), moringa (MLE) and mulberry leaf water extract (MBLE), T2 i.e. skipping the irrigation at grain filling stage and application of 2µM ABA, T3 i.e. skipping the irrigation at grain filling stage and application of 10 m mol SA, T4 i.e. skipping the irrigation at grain filling stage and application of 15% MLE and T5 i.e. skipping the irrigation at grain filling stage and application of 10% MBLE. The experiment was laid out in Randomized Complete Block Design with factorial arrangement and repeated three times. From this study it is concluded that Aas-2011 shown best result under drought condition by applying growth regulators and plant water extracts.

O fortalecimento de genótipos de trigo pela aplicação de reguladores de crescimento, solutos compatíveis e extratos vegetais sob restrição hídrica é uma importante estratégia para obtenção de produção sustentável. Trilha de campo foi realizada na área de pesquisa da Faculdade de Ciências Agrárias, Universidade de Ghazi, Dera Ghazi Khan, Paquistão. Três cultivares de trigo Aas-2011, Faisalabad-2008 e Triple anão-1 foram submetidas a estresse hídrico (pulando a irrigação no estágio de enchimento de grãos). Os genótipos de trigo foram submetidos a tratamentos, T1, ou seja, irrigação normal sem aplicação de ácido abscísico (ABA), ácido salicílico (SA), moringa (MLE) e extrato de água de amoreira (MBLE), T2¬, pular a irrigação em estágio de enchimento de grãos e aplicação de ABA 2µM, T3 ou seja, ignorando a irrigação no estágio de enchimento de grãos e aplicação de 10 m mol SA, T4 ou seja, ignorando a irrigação no estágio de enchimento de grãos e aplicação de 15% MLE e T5 ou seja, ignorando a irrigação no enchimento de grãos estágio e aplicação de 10% MBLE. O experimento foi exposto no delineamento de blocos completos casualizados com arranjo fatorial e repetido três vezes. A partir deste estudo conclui-se que Aas-2011 apresentou melhor resultado sob condição de seca, aplicando reguladores de crescimento e extratos de água de plantas.

Air-drying temperature changes the content of the phenolic acids and flavonols in white mulberry (Morus alba L.) leaves / Temperatura de secagem ao ar altera o teor de ácidos fenólicos e flavonóis em folhas de amoreira branca (Morus alba L.)

ABSTRACT: The white mulberry leaves are typically available on the market in dried or encapsulated form. It was assumed in the study that appropriate drying of leaves of the white mulberry is significant for obtaining intermediate products with high content of compounds having anti-oxidative activity. The purpose of the study was to determine the influence of the temperature of mulberry leaves air drying on the content of phenolic acids and flavonols. It has been determined that the content of these compounds in the leaves depended on the drying temperature. Drying at 60 °C favored release of phenolic acids and flavonols from complexes and/or formation of new compounds. Their total content was 22% higher than in leaves dried at 30 °C. Drying at 90 °C reduced the phenolic acid and flavonol content by 24%. The most favorable drying temperature was 60 °C.

RESUMO: As folhas da amoreira branca estão normalmente disponíveis no mercado em forma seca ou encapsulada. Assumiu-se no estudo que a secagem adequada das folhas da amora branca é importante para a obtenção de produtos intermediários com alto teor de compostos com atividade antioxidante. O objetivo do estudo foi determinar a influência da temperatura de secagem de ar de folhas de amoreira sobre o teor de ácidos fenólicos e flavonóis. Foi determinado que o conteúdo destes compostos nas folhas dependia da temperatura de secagem. Secagem a 60 °C favoreceu a liberação de ácidos fenólicos e flavonóis a partir de complexos e / ou formação de novos compostos. Seu teor total foi 22% superior ao das folhas secas a 30 °C. A secagem a 90 °C reduziu o teor de ácido fenólico e flavonol em 24%. A temperatura de secagem mais favorável foi de 60 °C.

Function analysis of anthocyanidin synthase from Morus alba L. by expression in bacteria and tobacco

Background: Flavonoids are a kind of important secondary metabolite and are commonly considered to provide protection to plants against stress and UV-B for a long time. Anthocyanidin synthase (ANS), which encodes a dioxygenase in the flavonoid pathway, catalyzes the conversion of leucoanthocyanidins to anthocyanidins, but there is no direct evidence indicating that it provides tolerance to stress in plants. Results: To investigate whether ANS can increase tolerance to abiotic stress, MaANS was isolated from mulberry fruits and transformed into tobacco. Our results suggested that the bacterially expressed MaANS protein can convert dihydroquercetin to quercetin. Overexpression of MaANS remarkably increased the accumulation of total flavonoids in transgenic lines and anthocyanins in corollas of flowers. Transgenic lines showed higher tolerance to NaCl and mannitol stress. Conclusions: These results indicated that MaANS participates in various dioxygenase activities, and it can protect plants against abiotic stress by improving the ROS-scavenging ability. Thus, this alternative approach in crop breeding can be considered in the improvement of stress tolerance by enriching flavonoid production in plants