The mammalian target of rapamycin (mTOR), is a serine/threonine protein kinase and belongs to the phosphatidylinositol 3-kinase (PI3K)-related kinase (PIKK) family. mTOR interacts with other subunits to form two distinct complexes, mTORC1 and mTORC2. mTORC1 coordinates cell growth and metabolism in response to environmental input, including growth factors, amino acid, energy and stress. mTORC2 mainly controls cell survival and migration through phosphorylating glucocorticoid-regulated kinase (SGK), protein kinase B (Akt), and protein kinase C (PKC) kinase families. The dysregulation of mTOR is involved in human diseases including cancer, cardiovascular diseases, neurodegenerative diseases, and epilepsy. Tissue damage caused by trauma, diseases or aging disrupt the tissue functions. Tissue regeneration after injuries is of significance for recovering the tissue homeostasis and functions. Mammals have very limited regenerative capacity in multiple tissues and organs, such as the heart and central nervous system (CNS). Thereby, understanding the mechanisms underlying tissue regeneration is crucial for tissue repair and regenerative medicine. mTOR is activated in multiple tissue injuries. In this review, we summarize the roles of mTOR signaling in tissue regeneration such as neurons, muscles, the liver and the intestine. Streptomyces hygroscopicus and called rapamycin [In the 1970s, a new antifungal, was discovered in soil samples on the Polynesian island of Rapa Nui, which was isolated from apamycin . Afterwaapamycin . During apamycin . Brown eapamycin . mTOR isapamycin . mTOR siapamycin . The ribapamycin . The mTOapamycin ,7. mTOR apamycin ,11,12. Tapamycin .Tissue damage caused by trauma, diseases, and aging, etc. can result in organ dysfunction. Afterward, tissue regeneration is critical for the restoration of organ functions and maintenance of homeostasis . In adulIn this review, we first briefly describe the structures, regulatory mechanisms, and physiological functions of mTORC1 and mTORC2. Then, we put our efforts toward summarizing the roles of mTOR signaling in the regeneration of neurons, muscles, the liver, and intestine. At the end, the development strategy of tissue-specific agonist or inhibitor of mTORC1 in regenerative medicine is discussed.RSK, both of which phosphorylate and inhibit TSC2 [mTOR is a serine/threonine protein kinase and a member of the PI3K-related kinase (PIKK) family, which forms the mTORC1 and mTORC2 complexes with other proteins ,18. The bit TSC2  , RAG GTPases  and Ragulator. Unlike other stimulators, mTORC1 activation by amino acids is independent of the TSC-Rheb signaling axis [The mTORC1 activation is closely related to the variation of amino acid concentrations. Different types of amino acid stimulate mTORC1 through different sensors. For example, cytosolic leucine, cytosolic arginine, and the lysosomal arginine are sensed by Sestrin2, CASTOR1 (Cellular Arginine Sensor for mTORC1) complex, and a candidate lysosomal amino acid sensor SLC38A9, respectively ,34,35,36ing axis   and 4E-BP1 . Then, tLike mTORC1, mTORC2 also contains mTOR and mLST8 subunits. But Raptor in mTORC1 is replaced by Rictor in mTORC2 . mTORC2 The mTORC2 mainly controls cell migration through phosphorylation of the AGC (protein kinase A/G/C) protein kinase family members such as PKCα , PKCδ 6, PKCξ 6, PKCγ anThe blood-brain barrier (BBB) is formed by endothelial cells, pericytes and astrocytes. These cells together form the neurovascular unit (NVU), which serves as an interface between the blood and the neural tissue. Impairment of BBB function is associated with neurodegenerative diseases . The braIn general, axons after injury do not spontaneously regenerate in adult mammalian CNS because of a diminished intrinsic regenerative capacity and extrinsic growth-inhibitory factors ,87,88. IAA can stimulate RGCs regenerative ability after optic nerve injury. This enhanced RGC regeneration is dependent on mTORC1 [Injuries to the optic nerves and spinal cord have been widely used to study axonal regeneration of the CNS. mTOR signaling exhibits distinct functions in the optic nerve and spinal cord regeneration . mTOR sin mTORC1 . Anothern mTORC1 . After on mTORC1 . Furthern mTORC1 ,109,110.Although studies above suggest that mTOR signaling promotes RGC regeneration, Li et al. showed that mTOR is a negative regulator of RGCs survival and activates astrocytes after optic nerve injury. In the rat retinal ischemia-reperfusion (I/R) injury model, neuroprotective effects of rapamycin can reduce the loss of RGCs after optic nerve injury. Thus, the neuroprotective effects of rapamycin supply a therapeutic method for optic neurodegeneration . OverallThe spinal cord connects the brain with the PNS. Spinal cord injuries (SCI) are divided into traumatic and non-traumatic aetiologies . MechaniMuscle can quickly regenerate to recover its function after nearly complete myofiber destruction. It is important to maintain the physiological homeostasis of muscle throughout life . Many stmtor or raptor in MuSCs effectively inhibits activation, proliferation, and differentiation of satellite cells, which impairs skeletal muscle regeneration [myogenin and stimulates MuSCs differentiation into myoblasts [rictor knockout in embryonic and adult satellite cells is ineffective in skeletal muscle regeneration [+ T cells rather than rictor−/− CD4+ T cells promote muscle regeneration in the Rag1−/− mice [In mice, conditional knockout (cKO) of neration ,136,137 neration ,136. Rapneration . S6K1 isneration . Ablatioyoblasts   , BaCl2 [2  and isch2 , some pa2 . This reneration . The IGFneration ,149. mTOreatment . Furtherreatment .Muscle-mass weakness and wasting are often caused by various pathological conditions, such as sarcopenia, diabetes and chronic obstructive pulmonary disease (COPD). On average, geriatric patients with sarcopenia lose 30% of their muscle mass and 35% of myofibers , and hav4)- and thioacetamide (TAA)-induced chemical liver injury. Remaining hepatocytes after PH or chemical liver injury accomplish liver regeneration via self-replication without the participation of progenitor cells [The liver possesses extraordinary regenerative capacity comparing to other internal organs ,168,169.or cells . Howeveror cells . After eor cells ,175 and or cells ,177 , one of the sirtuin family of class III NAD+-dependent histone deacetylase, delays G1 to S phase transition and impairs activation of the Akt/mTOR pathway during liver regeneration [let-7 deletion [A 2/3 hepatectomy in rodents promotes the release of cytokines and growth factors , which sreatment . mTOR acreatment   can also be important for liver regeneration ,179,200.In addition to endogenous liver regeneration, liver transplantation is also an effective way to restore liver mass and functions after liver failure or cirrhosis ,203. In + (escargot) intestinal stem cells (ISCs) and enteroblasts (EBs) are responsible for gut homeostasis and drive the regeneration following injury [Lgr5 marks the ISCs at the base of intestinal crypt which maintain the intestinal homeostasis and intestinal regeneration in mouse [Dll1+ secretory progenitor cells, Alpi+ enterocyte progenitor cells, and Lyz+ Paneth cells can be induced to ISC after specific ablation of Lgr5+ ISC [The small intestinal epithelium is a single cell layer with rapid self-renewing and strong regenerative capabilities. The mammalian intestinal epithelium mainly includes enterocytes, Goblet cells, enteroendocrine cells, and Paneth cells . In flieg injury ,209. Theg injury ,210. Alsg injury . Similarin mouse . Furthergr5+ ISC ,214,215.gr5+ ISC . In addigr5+ ISC . Taken tDrosophila and mammals. In inflammatory bowel disease (IBD) mouse models induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) or dextran sodium sulfate (DSS), mTORC1 is required for intestinal regeneration against IBD. The deficiency of Regnase-1 promotes the intestinal regeneration in the acute IBD via controlling the mTOR and purine metabolism [The intestinal epithelium, which has a powerful regenerative capacity after injury, provides a model to study ISCs, cancer, and intestinal regeneration . A few stabolism . Akt/mTOtabolism ,220. mTOtabolism . These stabolism . Besidestabolism .+ memory T cells. But in this process, sirolimus causes reversible oral ulcers, headaches, and cytopenia [In this review, we summarize the roles of mTOR in the regeneration of neurons, muscles, the liver, and the intestine, which are mainly mediated by mTORC1 rather than mTORC2 signaling. The mTORC1 signaling network may be disrupted in various diseases, such as ALS, AD, PD, COPD, sarcopenia, liver failure, and IBD, which is harmful to tissue regeneration ,225,226.ytopenia . The tis


Nemopilema nomurai is one of the most perilous organisms and leads to significant deleterious outcomes such as harm to the fishery, damage the coastal equipment, and moreover, its envenomation can be hazardous to the victims. Till now, the components of Nemopilema nomurai venom (NnV) are unknown owing to scant transcriptomics and genomic data. In the current research, we have explored a proteomic approach to identify NnV components and their interrelation with pathological effects caused by the jellyfish sting. Altogether, 150 proteins were identified, comprising toxins and other distinct proteins that are substantial in nematocyst genesis and nematocyte growth by employing two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI/TOF/MS). The identified toxins are phospholipase A2, phospholipase D Li Sic Tox beta IDI, a serine protease, putative Kunitz-type serine protease inhibitor, disintegrin and metalloproteinase, hemolysin, leukotoxin, three finger toxin MALT0044C, allergens, venom prothrombin activator trocarin D, tripeptide Gsp 9.1, and along with other toxin proteins. These toxins are relatively well characterized in the venoms of other poisonous species to induce pathogenesis, hemolysis, inflammation, proteolysis, blood coagulation, cytolysis, hemorrhagic activity, and type 1 hypersensitivity, suggesting that these toxins in NnV can also cause similar deleterious consequences. Our proteomic works indicate that NnV protein profile represents valuable source which leads to better understanding the clinical features of the jellyfish stings. As one of the largest jellyfish in the world, Nemopilema nomurai sting is considered to be harmful to humans due to its potent toxicity. The identification and functional characterization of its venom components have been poorly described and are beyond our knowledge. Here is the first report demonstrating the methodical overview of NnV proteomics research, providing significant information to understand the mechanism of NnV envenomation. Our proteomics findings can provide a platform for novel protein discovery and development of practical ways to deal with jellyfish stings on human beings.Nowadays, proliferation of jellyfish has become a severe matter in many coastal areas around the world. Jellyfish  N. nomurai jellyfish accidents have been reported in the coastal areas of China, Korea, and Japan, and life-threatening cases were also observed in humans [N. nomurai known as the giant jellyfish is one of the most dangerous species belonging to the Phylum Cnidaria, being their diagnostic feature presence of stinging organelles called nematocysts, located mostly on the tentacles of jellyfish [Over the recent decades, there is a huge expansion of jellyfish blooms worldwide, which cause severe damage to the fishery and disturb marine ecosystem . Since 1n humans . N. nomuellyfish ,4. On suellyfish ,6,7,8. Eellyfish ,10. In tIn many venomous creatures such as snakes, spiders, scorpions, and cone snails, proteomic approaches have been auspiciously exploited to purify and characterize their venom components ,12,13,14N. nomurai jellyfish have shown high 71 sequence similarity with those of other venomous and poisonous animals. Mainly, it is composed of Phospholipase A2, Phospholipase D Li Sic Tox beta IDI, Serine protease, Putative Kunitz-type Serine protease inhibitor, Disintigrin and Metalloproteinase, Hemolysin, Leukotoxin, Three-finger toxin MALT0044C, allergens, Venom prothrombin activator trocarin D, Tripeptide Gsp 9.1 and Cell death abnormality protein 1 from Caenorhabditis elegans. Several toxin proteins shared homology with proteins from microorganisms such as RTX-III toxin determinant A from serotype 2 from Actinobacillus pleuropneumoniae, Shiga-like toxin 1 subunit A from Enterobacteria phage, Leukotoxin from Pasteurella haemolytica, Cell death abnormality protein 12 from Saccharomyces cerevisiae, and Pro-apoptotic serine protease nma111 from Neurospora crassa. Interestingly, it also includes non-toxic proteins during NnV profiling, such as PCNA-interacting partner, Fukutin, Cell death abnormality protein 1, UvrABC system protein A, Division control protein 7, and OTU domain-containing protein 7B.The identification and functional characterization of NnV components have been poorly characterized. Herein, we described the proteomic profile of the NnV. Our present study has successfully demonstrated the proteomic characterization of NnV by utilizing 2-DE and MALDI/TOF/MS. 2-DE gel electrophoresis, revealing the venom components ranging from PI 3–10 and molecular weights between 15–250 kDa, as shown in N. nomurai jellyfish venom proteolytic activity using various types of substrates such as gelatin, casein, and fibrin, as shown in N. nomurai jellyfish venom possesses higher gelatinolytic, caseinolytic, and fibrinolytic behavior. The gelatinolytic activities of NnV showed the greatest proteolytic activity as comparision to casein and fibrin zymography and displayed different protein banding pattern between 150–25 kDa. The majority of the gelatinolytic activities were inhibited in the presence of broad-spectrum metalloproteinase inhibitor , confirming that metalloproteinase-like enzymes are present in N. nomurai jellyfish venom. Furthermore, casein zymography showed strong enzymatic activity, which could be evaluated in between the range of 70–20 kDa. In fibrin zymography, weaker fibrinolytic activity was observed at 70–25 kDa. The caseinolytic and fibrinolytic activities disappeared in the presence of 1,10-phenanthroline. Despite this, we have performed 2-DE zymography under the non-reducing condition to determine the proteolytic patterns, as shown in Zymography assay was performed to identify the All identified proteins were subsequently classified into four ontologies according to their molecular functions, biological processes, cell components, and protein classes. In the molecular function classification, the most abundant protein category is of catalytic activity (46%) and binding (42%). Few proteins are associated with transporter activity (5%), receptor activity and structural molecular activity (3%) and translation regulator activity (1%). For the molecular function catalytic and binding activities designated the first rank, which can be related to the toxins components in NnV and hint towards strong toxicity. In the biological process category, the two significant groups include a cellular process (31%) and the metabolic process (28%). Followed by the response to stimulus (8%), developmental process (7%), biological regulation (5%), immune system process (5%), localization (5%), and multicellular organismal process (4%). The small number of proteins are assigned with biological adhesion (3%) and reproduction (1S%), as shown in Over the last few decades, jellyfish envenomation has turned into a universal health issue and gave rise to approximately 150 million envenomation cases yearly . JellyfiCrotalus durissus snake species, A Disintegrin and Metalloproteinase with thrombospondin motifs 10, A Disintegrin and Metalloproteinase with thrombospondin motifs 23 from Mus musculus, along with ATP-dependent Zinc Metalloprotease FtsH from Oenococcus oeni. In the present study, our proteomic analysis showed that metalloproteinase are the second most abundant components of NnV, comprising 21% of venom proteome, as shown in Metalloproteases are well known for hemorrhagic activities such as fibrinolysis, apoptosis, inhibit platelet aggregation, interfere with blood coagulation, cause pro-inflammatory activity and deactivate blood serine protease inhibitors . Our proNemopilema nomurai, Cyanea nozakii, Rhopilema esculenta, and Aurelia aurita contains various Metalloproteinases [Nemopilema nomurai might be responsible for swelling, inflammation, and dermonecrosis [Nemopilema nomurai were also reported to induce dermal toxicity in both the in-vivo and in-vitro animal model [Moreover, in our previous study, we have found that the venoms of the scyphozoan jellyfish species including teinases , which steinases . Therefonecrosis . Previoual model . In futuLoxosceles intermedia spider venom, 85/88 kDa Calcium-independent Phospholipase A2 and Phospholipase A-2-activating protein from Rattus norvegicus, Calcium-independent Phospholipase A2-gamma from Mus musculus. Phospholipase D LiSicTox-betaIDI is the novel member of loxosceles dermonecrotic toxin family, which has diverse biological activities such as dermonecrosis, increased vessel permeability, platelet aggregation, induce an inflammatory response, and cause mortality in animal models [Several phospholipases were also identified in NnV in current research, such as Phospholipase D LiSicTox-betaIDI, also known as LiRecDT5 homologous to PLA2 from l models ,24. The Vipera nikolskii, serine protease HtrA-like from Staphylococcus aureus (strain NCTC 8325), Pro-apoptotic serine protease nma111 from Neurospora crassa (strain ATCC 24698/74-OR23-1A/CBS 708.71/DSM 1257/FGSC 987). Snake venom Serine Protease Nikobin can acts on the hemostasis system of the prey by affecting several physiological processes that include blood coagulation, fibrinolysis, and blood pressure [Proteases constituted a vital portion of NnV counting 5% of the venom proteome . Serine pressure . AccordiPutative Kunitz-type Serine protease inhibitor is efficiently characterized in well-known venomous animals, for example snakes, spiders, scorpions, centipedes, cone snails, and sea anemone ,37,38,392+ channels [Other toxin protein types present in NnV are homologous to the three-finger toxin MALT0044C, hemolysins, leukotoxins, major pollen allergen Lol p 5a, Allergen Mag, venom prothrombin activator trocarin D, RTX-III toxin determinant A from serotype 2, and Shiga-like toxin 1 subunit A. Three-finger toxin MALT0044C belongs to the snake three finger toxin family, it has a predominant role in neurotoxicity by inducing peripheral paralysis and finally leads to respiratory arrest and death . The prochannels . TherefoS. meleagris [C. fleckeri venom, which share sequence homology to toxins related to 4 cubozoan jellyfish species [C. alata, CrTX-1 from C. rastoni, and CcTX-1 from C. capillata were also reported in previous studies [Various types of Hemolysins were reported in the venom of different kinds of jellyfish, including Ryncolin-2, Ryncolin-3, Neoverrucotoxin subunit beta, and Veficolin-1 were identified in the venom of jellyfish eleagris . Moreove species . Other h studies ,47,48,49 studies ,47,48,49Serratia marcescens. Hemolysin triggers the release of the inflammatory mediators, increase vascular permeability, cause edema formation, granulocyte accumulation, and finally contribute pathogenicity of Serratia species [Hemolysin transporter protein was also identified in NnV, which is homologous to ShlB of  species ,51. TherPasteurella Leukotoxins are exotoxins that attack host leukocytes, chiefly polymorphonuclear cells, by inducing cell rupture [2+, finally results in host cell rupture [Pasteurella Haemo-lytica-like sp. (strain 5943b), which may play a significant role in NnV envenomation and cause harmful consequences. rupture . The Leu rupture . We alsoLolium perenne and Allergen Mag (fragment) from Dermatophagoides farina. Lol p 5a is a major allergen of rye-grass pollen (Lolium perenne pollen), which mediate type I hypersensitivity, cause an allergic reaction in humans, hay fever, and triggers allergic asthma. House dust fly (Dermatophagoides farina) is the one of the major agent causing allergic ailment by way of bronchial, asthma, atopic dermatitis and rhinitis [Dermatophagoides farina, which binds to IgE and causes an allergic reaction in humans [Two types of allergens were also detected in the NnV proteomics known as Major pollen allergen Lol p 5a from rhinitis ,54. Allen humans ,56,57. ITropidechis carinatus, acts as a toxin component of the snake venom and disturbs the hemostatic system of prey [Tropidechis carinatus. Therefore, venom prothrombin activator trocarin D is one of the important toxic protein in NnV which might cause severe harmful consequences such as shock, cardiovascular instability, and sudden death. RTX-III toxin determinant A from serotype 2 and RTX-I toxin determinant A from serotypes 1/9 are an essential virulence factor for A. pleuropneumoniae and cause swine pleuropneumonia [Venom prothrombin activator trocarin D is the snake prothrombin activator from  of prey . It was  of prey . Fortunaneumonia .RTX-III PROTEIN are toxic to porcine lung macrophages and erythrocytes, and it possesses strong hemolytic activity and causes cytotoxicity in alveolar macrophages and neutrophils hence play a significant role in pathogenesis . NoteworIt was reported earlier that a Shiga-like toxin subunit A negatively regulates the translation process by catalytic inactivation of 60S ribosomal subunit and responsible for inhibiting protein synthesis . The shiCaenorhabditis elegans, was also identified in NnV, and Cell death abnormality protein 1 necessitates the engulfment of cells undergoing programmed cell death [Surprisingly, along with toxic proteins, we have also recognized many nontoxic proteins during NnV profiling, which plays an important role in signaling and other metabolic processes. Kinases itself constitute the major part of NnV, they hold 22% of known NnV proteins. Other proteins are 30% of total proteins found in NnV. OTU domain-containing protein 7B is one of the nontoxic proteins found in NnV. OTU domain-containing protein 7B is the negative regulator of the non-canonical NF-kappa-B pathway and displays the anticancer effect by suppressing the NF-kappa-B pathway in HCC cells . OTU domll death ,67. It all death . HoweverNemopilema nomurai is one of the largest jellyfish and its evenomation can lead to harmful consequences, even death. Till now the identification and functional characterization of NnV components have been poorly described and remain beyond our knowledge. This is the first report that is able to identify around 150 proteins in Nemopilema nomurai jellyfish venom, which include metalloproteinases, kinases, phospholipases, proteases, toxins, and allergens. Some of these components are considered to play an important role in the cardiovascular, hepatotoxic, hematological, cytotoxic, and allergenic effects of the venom. These findings are beneficial in recognizing the mechanism of jellyfish envenomation and provide valuable means to establish new methods or treatments to handle jellyfish stings. The overall comprehensive characterization of NnV, provide a promising approach to explore the rich source of bioactive toxin components as therapeutic agents in the future.Jellyfish Immobiline™ Drystrip , dithiothreitol (DTT), and iodoacetamide were bought from GE Healthcare life sciences . Acetonitrile (ACN), trifluoroacetic acid was purchased from Merck Chemicals, Darmstadt, Germany. Sequencing grade modified trypsin was purchased from Promega Corporation, Madison, WI, USA, formic acid from Acros Organics BVBA, Geel, Belgium. Other analytical grade chemicals and reagents were used and purchased from Sigma Aldrich Corporation .N. nomurai jellyfish specimens were collected from the Yellow Sea near the coast of Gunsan, South Korea. After taking out of water tentacles were dissected and transferred immediately in ice to the laboratory for further processing. Isolation of nematocysts was done using the previously described method [v/v) of cold sea water were added and at 4 °C put on shaker for one day. After that, the tentacle-free seawater was harvested and centrifuged at 1000× g for 5 min; then the nematocyst-rich pellet was washed thrice with fresh sea water. The remaining sediment tentacles were later autolyzed in fresh seawater at 4 °C for one day, as described above, and the autolysis process was repeated for 3–4 days. At last, the nematocysts that settled down were collected and seawater was used to wash them several times. Later, the nematocysts were centifuged at 500× g for 5 min, and pellets (nematocysts) were lyophilized and stored at −20 °C until further use.d method . In shorThe technique described by Carrette and Seymour was used to extract venom from the freeze-dried nematocysts  with sliw/v) CHAPS, 10 mg/mL DTT, 1% pharmalytes 3–10 and few grains of bromophenol blue). Then, protein samples were applied to immobiline TM Dry strip 18 cm, pH gradient (IPG) strips (pH 3–10) and passively rehydrated overnight at room temperature. The isoelectric focusing (IEF) was performed at a constant temperature of 20 °C using Ettan IPGphor system , with the following procedure: 50 V for 1:00 h, 200 V for 1:00 h, 500 V for 0:30 h, gradient 4000 V for 0:30 h, 4000 V for 1:00 h, gradient 10,000 V for 1:00 h, 10,000 V for 13:00 h, and 50 V for 3:00 h. Before the second dimension, the focused strips were firstly reduced with equilibration buffer (50 mM Tris-HCl (pH 8.8), 6 M urea, 30% glycerol, 2% SDS and 0.01% bromophenol blue containing 1% w/v DTT) for 15 min. For the second time, the gel strips were washed with the same equilibration buffer containing 2.5% w/v iodoacetamide for 15 min. The focused and equilibrated strips were inserted on top of the 12% SDS-PAGE and sealed with 0.5% w/v agarose gel. After that, electrophoresis was performed in a PROTEAN II xi cell gel electrophoresis unit. The gels were run at 10 mA/gel for 15 min for the initial migration and increased to 20 mA/gel at 20 °C for separation until the dye front reached the bottom of the gels. Silver staining was performed to visualize the protein spots as similar to the method reported by Mortz et al. [For two-dimensional gel electrophoresis, 500 µg of protein samples were resuspended in 2-DE sample buffer  for 10 min with shaking. Further, the gel pieces were dehydrated for 10 min by incubating in 100 µl of 100% acetonitrile and were then dried in a lypholizer equipment for 15 min. The reduction step was performed by adding 50 µL reduction solution (10 mM DTT in 100 mM ammonium bicarbonate) to the dried gel pieces and incubated for 45 min at room temperature. After that, the gel pieces were alkylated with 100 mM iodoacetamide in the dark for 45 min at room temperature. The gel pieces were dried, incubated in trypsin  at a final concentration of 2 ng/µL in 10 µL of 50 mM NH4HCO3 and incubated on ice for 45 min. The excessive liquid was removed, and the proteolysis of protein was performed by adding the exact amount of 50 mM NH4HCO3. After overnight digestion with trypsin at 37 °C, the tryptic peptide mixture was pooled with extraction buffer containing 100% acetonitrile and 50% trifluoroacetic acid and concentrated in a speed vacuum.In-gel digestion of the proteins was performed by the method described previously . The proThe peptide samples were mixed with 1 µL of HCCAs matrix solution (α-acyano-4-hydroxycinnamic acid) and 1 µL of extraction buffer. 1 µL peptide mixture was spotted onto a freshly cleaned MALDI/TOF plate and dried at room temperature. Mass spectra were measured by using Voyager-DE STR mass spectrometer .http://prospector.ucsf.edu), and Mascot (Matrix science http://www.matrixscience.com) in the Swiss-Prot databases. For the peptide search following parameters were considered carbamidomethylation of cysteines as a fixed modification, oxidation of methionine as a variable modification, peptide mass tolerance of ±30 ppm for the fragment ions, trypsin with one missed cleavage was allowed. Criteria used for protein identification includes a number of the matched peptide, the extent of sequence coverage and probability based Mowse score was considered before accepting the identification.Reflection/delayed extraction mode acquired the spectra. Monoisotopic peptide masses were chosen over a mass range of 800–3000 Da. Peptide mass fingerprinting (PMF) was performed to identify proteins using the MS-Fit program , 140 mM NaCl, 2.5 mM CaCl2, 2.4% human erythrocytes, and 2% egg yolk. After overnight incubation at 37 °C, the opaque zone indicated the presence of PLA2 [NnV was run under non-reducing conditions on 12% SDS-PAGE and stained with Coomassie Brilliant Blue R-250. Blood and egg yolk zymography method  with sli of PLA2 .w/v) CHAPS, 1% pharmalytes 3–10 and few grains of bromophenol blue) without DTT and the strips were rehydrated overnight, and the isoelectric focusing was performed as the method described above. After IEF the focused strips were washed with equilibration buffer (50 mM Tris-HCl (pH 8.8), 6 M urea, 30% glycerol, 2% SDS, and 0.01% bromophenol blue) for 15 min followed by electrophoresis. For 2-DE gelatin zymography 12% SDS-polyacrylamide gels were copolymerized with gelatin (1 mg/mL), and electrophoresis was performed at 4 °C. The gels were washed twice after electrophoresis for 30 min in 2.5% Triton X-100 to remove SDS. Triton X-100 traces were removed from the gels by washing with deionized water. After that, the gels were incubated in 20 mM Tris (pH 7.4), 0.5 mM calcium chloride at 37 °C for 16 h and stained with 0.125% coomassie blue. Clear zones of hydrolysis in the gel against blue background showed gelatin proteolytic activity.The NnV samples were prepared under non reducing conditions for 2-DE and gelatin 2-DE zymography. The 500 µg of protein samples were dissolved in 7 M urea, 2 M thiourea, 4% (http:www.pantherdb.org/) to classify regarding molecular function, biological process, protein class, and cellular components [The proteomic data was analyzed using the Panther classification system (mponents .


Eriobotrya japonica is traditionally used as an antipyretic, digestive, and diuretic agent. Its flowers possess free radical–scavenging, antioxidative, and hepatoprotective effects. We investigated the hepatoprotective potential of E. japonica leaf extract and its various fractions against hepatotoxicity in rats. Liver injury was stimulated by the oral administration of carbon tetrachloride . Male albino rats (n = 55) were distributed arbitrarily into 11 groups: Group I, normal control group; Group II, CCl4 (positive control group); Group III, CCl4 + silymarin; Groups IV and V, CCl4 + two doses of 250 and 500 mg/kg of the 80% methanolic extract of E. japonica leaves, respectively; Groups VI and VII, CCl4 + 250 mg/kg and 500 mg/kg of the ethyl acetate fraction, respectively; Groups VIII and IX, CCl4 + 250 and 500 mg/kg of the butanol fraction, respectively; and Groups X and XI, CCl4 + 250 and 500 mg/kg of the aqueous fraction of E. Japonica leaves, respectively. CCl4-treated rats that were given 250 or 500 mg/kg of the methanol extract of E. Japonica leaves, or its ethyl acetate, butanol, or aqueous fractions, had significantly lower levels of biochemical parameters such as alanine aminotransferase, aspartate transaminase, alkaline phosphate, total protein, gamma-glutamyl transferase, and bilirubin levels than those of the CCl4 positive group. However, the extract and fractions did not significantly affect lipid profiles. Thus, we conclude that Eriobotrya leaf extract and its fractions have a hepatoprotective effect against CCl4-induced hepatotoxicity in rats. Eriobotrya japonica, commonly known as loquat, an evergreen plant that belongs to the family Rosaceae.Medicinal plants and their extracts represent a rich source of crude medications that possess therapeutic properties. Indeed, the World Health Organization reports that various plant fractions and their dynamic constituents are utilized as traditional medicines by 80% of the world population . One sucEriobotrya japonica originated in southeast China, but it is now grown in India, Japan, Korea, and other countries. It is small in size, with narrow leaves that are light in color on the underside and darker green on the upper surface. The leaves are valuable for the treatment of incessant maladies such as dysmenorrhea, lower back pain, asthma, chronic bronchitis, asthma, phlegm, and headache [headache . In Japaheadache . E. japonica. Several of those compounds are active against viruses, inflammation, hyperglycemia, and tumors [ E. japonica leaves are tormentic acid, oleanolic acid, maslinic acid, ursolic acid, and corosolic acid [A variety of natural compounds such as tannins, megastigmane glycosides, triterpenes, sesquiterpenes, and flavonoids have been found in the leaves ofd tumors , 4–6. Splic acid . Previoulic acid , 9. E. japonica flowers possess free radical–scavenging, antioxidative, and hepatoprotective effects against mercuric chloride induced hepatotoxicity in rats [The hydroalcoholic extracts of in rats . E. japonica, we injected male rats with carbon tetrachloride (CCl4), an extremely lethal chemical compound that is commonly used for inducing hepatotoxicity in animal studies [ E. japonica leaves, as well as its ethyl acetate, butanol, and aqueous fractions, against CCl4-induced hepatotoxicity were investigated. In this way, we aimed to scientifically validate and support the conventional uses and previous findings of E. japonica. To explore the possible scavenging capacity and hepatoprotective effects of studies , 12. The E. japonica were collected from Wadi Houf, Helwan, Egypt, and taxonomically confirmed by Professor Ibrahim Elgarf, Professor of Taxonomy, Faculty of Science, Cairo University, Cairo, Egypt.In April 2016, the leaves ofDried shade leaves were powdered (1000 g) and extracted with 80% (v/v) methanol and then vacuum-concentrated under reduced pressure at 40 ± 1°C until dry. The residue was designated as methanolic extract (Er-1). A portion of Er-1 (70 g) was dissolved in water and fractionated successfully with hexane, chloroform, ethyl acetate (Er-2), and butanol (Er-3); the remaining aqueous solution was designated as Er-4.4, isoflurane, and other chemicals were procured from El-Gomhouria Company, Cairo, Egypt. Kits used for biochemical analysis of alkaline phosphate (ALP), alanine aminotransferase (ALT), aspartate transaminase (AST), total protein, gamma-glutamyl transferase (GGT), bilirubin, cholesterol, triglycerides, high density lipoprotein (HDL), malondialdehyde (MDA), nonprotein sulfhydryl groups (NP-SH), and total protein were obtained from the Gamma Trade Company for Pharmaceutical and Chemicals, Dokki, Egypt.Antioxidants, formalin, CCl4 (2 mL/kg) [Liver damage was induced by subcutaneous injections of CCl2 mL/kg) . ad libitum, and the rats were fed a basal diet for the entirety of the study. Afterward, 55 male rats  were randomly divided into 11 groups as follows. In Group I, rats were subcutaneously injected with paraffin oil and then marked as the normal control group. In Group II, rats were subcutaneously injected with CCl4 (2 mL/kg b.wt.) and then marked as the positive control group. In Group III, rats were subcutaneously injected with CCl4 (2 mL/kg b.wt.) and silymarin 10 mg/kg. Silymarin is the most used natural constituent for the healing of hepatic diseases worldwide due to its antifibrotic, anti-inflammatory, and antioxidant activities. Silymarin functions by stabilizing biological membranes and increasing protein synthesis [Rats were acclimated to animal laboratory conditions at 25°C, 55% humidity, and a 12 h:12 h light-dark cycle for seven days prior to testing. Water was suppliedynthesis . Therefo4 (2 mL/kg b.wt.) and Er-1 . In Groups VI and VII, rats were subcutaneously injected with CCl4 (2 mL/kg b.wt.) and Er-2 . In Groups VIII and IX, rats were subcutaneously injected with CCl4 (2 mL/kg b.wt.) and Er-3 . In Groups X and XI, rats were subcutaneously injected with CCl4 (2 mL/kg b.wt.) and Er-4 . Experiments were performed in Biology Lab of Medicinal Aromatic and Poisonous Plants Research Center College of Pharmacy King Saud University, Saudi Arabia, April-June 2018.In Groups IV and V, rats were subcutaneously injected with CClAt the end of four weeks, food (but not water) was withheld from all animals for 12 h. All rats were sacrificed with isoflurane. Blood samples were collected in clean centrifuge tubes via cardiac puncture. The samples were centrifuged at 3000 rpm for 15 min to separate the serum. The serum was carefully removed and transferred into lavender test tubes and solidified at 20°C until utilization for biochemical experiments.The livers of all test rats were divided into two; one half was immediately used for biochemical analysis, and the second half was subjected to histopathology examination.Biochemical parameters such as ALT, AST, GGT, bilirubin, and ALP were calculated with the help of Reflotron Plus analyzer and commercially available kits (Roche) . Commercμm thick segments were primed and stained with hematoxylin and eosin [All animals were euthanized under isoflurane anesthesia and then sacrificed. Small samples of liver, kidney, and testis were immediately sliced. The sections were fixed in 10% formol saline for 24 h and then bathed in tap water, dried in increasing concentrations of ethanol, cleaned in xylene, and inserted in paraffin wax (melting point of 55–60°C). Then, 6 nd eosin .Data are shown as the mean ± standard error. The level of statistical significance  among the groups was determined by one-way analysis of variance (ANOVA) followed by Dennett's multiple comparison test.4 group than in the control group. Those levels were considerably lower in the silymarin group than in the CCl4 group. Administration of 250 and 500 mg/kg of Er-1, Er-2, Er-3, and Er-4 in CCl4-treated rats significantly reduced the levels of ALT, AST, ALP, GGT, and bilirubin compared to those of the CCl4 positive control group . Pretreatment with silymarin at 10 mg/kg significantly reduced MDA to levels that nearly matched those of the control group. Although the MDA levels of CCl4-treated rats that were preadministered low dose (250 mg/kg) Er-1 did not significantly differ from those of the CCl4 positive control group, the MDA levels of CCl4-treated rats in preadministered high dose (500 mg/kg) of Er-1 were significantly lower than those of the CCl4 positive control group (P < 0.001). Postreatment of CCl4-treated with low- or high dose Er-2 significantly reduced MDA levels as compared to those of the CCl4 positive control group (P < 0.001). Similarly, treatment of CCl4-treated rats with low- or high dose Er-3 significantly reduced MDA levels as compared to those of the CCl4 positive control group (P < 0.001). Whereas rats treated with low dose Er-4 did not have significantly lower MDA levels than those in the CCl4 positive control group, those treated with high dose Er-4 experienced a significant reduction in MDA levels (MDA levels indicated that lipid peroxidation was significantly higher in the CClA levels .4-treated rat's preadministered low (250 mg/kg) or high (500 mg/kg) doses of Er-1 or Er-4, there was insignificant recovery of NP-SH levels as compared to those of the CCl4 group. However, samples of Er-2 or Er-3 at doses of 250 or 500 mg/kg caused significant increases in NP-SH levels as compared to those of the CCl4 positive control group  Er-1, the hepatocytes did not completely recover; a portion of their livers had mild cellular swelling and moderate hepatocyte necrosis (4 and presupplemented with high dose (500 mg/kg) of Er-1 had normal hepatic lobules, but their hepatocytes were not normal  of Er-2. Their liver slices revealed slight cellular swelling and moderate focal necrosis of hepatocytes (4-treated rats supplemented with high dose (500 mg/kg) of Er-2 showed normal hepatic lobules and normal hepatocytes  of Er-3 showed abnormal hepatocytes, with mild cellular swelling and focal necrosis that is associated with inflammatory infiltration  of Er-4 showed distorted architecture of the hepatic lobules that is associated with mild hepatocytic swelling (4-treated rats that were administered high dose (500 mg/kg) of Er-4, the scale of liver injury was miniscule and hepatic lobules were normal; however, fatty changes were observed  bind covalently to macromolecules and induce peroxidative degradation of membrane lipids of endoplasmic reticuli rich in polyunsaturated fatty acids. This process leads to excessive formation and accumulation of lipids in tissues such as the liver. In addition, lipids from peripheral adipose tissue are translocated to the liver for accumulation [4 treatment reduced HDL levels and raised serum cholesterol, triglyceride, and LDL levels, which possibly reflects impairment of the liver cells' ability to metabolize lipids and transform cholesterol to bile acid for excretion. Eriobotrya japonica extract and its various fractions attenuated those effects.Studies have shown that trichloromethyl radicals of CClmulation , 24. In 4 raised the levels of SGPT, SGOT, ALP, and bilirubin in rat serum. Such enzymes are normally present at higher levels in cell cytoplasm than in the serum. Hepatopathy causes release of AST, ALT, and bilirubin into the bloodstream in proportion to the extent of liver damage [4-treated rats indicated cellular damage and loss of cell membrane function [4-induced elevation of SGPT, SGOT, ALP, and bilirubin levels. The effects of the leaf formulations were possibly due to the presence of flavonoids.Our results showed that CClr damage . In our function . This waStudies , 27 have4 causes disruption and disassociation of polyribosomes on the endoplasmic reticulum, which leads to reduction of protein biosynthesis [4 caused depletion of total proteins, which indicates liver toxicity [4-induced total protein depletion, administration of Er-1 or Er-4 did not.CClynthesis . In our toxicity . Whereas4 positive control group had significantly lower NP-SH levels than did the normal control group. Administration of 10 mg/kg silymarin significantly reduced NP-SH levels below those of the CCl4 positive control group. The recovery of the NP-SH levels after treatment with silymarin was closer to that observed in the normal control group. Furthermore, whereas administration of Er-2 or Er-3 significantly attenuated the CCl4-induced reduction of NP-SH levels, administration of Er-1 or Er-4 did not  [Many researchers suggested the proposed mechanism is reducing proinflammatory mediators or increasing anti-inflammatory cytokine (such as IL-10) discharges are significant mechanisms for the anti-inflammation effects of loquat extracts , 18–21.  (NF-κB) , 22–24 a (NF-κB) , 22, 23, (NF-κB) , 35–39The relative levels of hepatoprotection afforded by the extract and fractions were as follows: Er-2 > Er-3 > Er-4 > Er-1.4-induced pathological changes in the liver. Er-2 and Er-3 are more effective at protecting the liver from damage than Er-1 and Er-4 and recommended for further bioscreening natural products.The above results suggest that Er-1, Er-2, Er-3, and Er-4 have hepatoprotective activity and that such activity is conferred through their ability to reduce the lipid peroxidation rate, promote antioxidant effects, and inhibit CCl


Context:Scutellaria baicalensis Georgi (Lamiaceae) is a popular medicinal plant. Its roots are used as the famous traditional Chinese medicine Huang-Qin, which is recorded in Chinese Pharmacopoeia, European Pharmacopoeia, and British Pharmacopoeia.Objective: This review comprehensively summarizes research progress in phytochemistry, pharmacology, and flavonoid biosynthesis of S. baicalensis.Methods: English and Chinese literature from 1973 to March 2018 was collected from databases including Web of Science, SciFinder, PubMed, Elsevier, Baidu Scholar (Chinese), and CNKI (Chinese). Scutellaria baicalensis, chemical constituents, phytochemistry, biological activities, and biosynthesis were used as the key words.Results: A total of 126 small molecules (1–126) and 6 polysaccharides have been isolated from S. baicalensis. The small molecules can be classified into four structural types, namely, free flavonoids, flavonoid glycosides, phenylethanoid glycosides, and other small molecules. Extracts of S. baicalensis and its major chemical constituents have been reported to possess anti-viral, anti-tumor, anti-bacterial, antioxidant, anti-inflammatory, hepatoprotective, and neuroprotective activities. Key steps in the biosynthetic pathways of Scutellaria flavonoids have also been summarized.Conclusions: This article could be helpful for researchers who are interested in the chemical constituents, bioactivities, biosynthesis, and clinical applications of S. baicalensis. Scutellaria L. (Lamiaceae) are perennial herbs with around 360 species in the world. Many of these species have medicinal uses , one of the most popular traditional Chinese medicines  in around 200 AD. In ancient Chinese language, the character ‘Qin’ means ‘herb for hemostasis’, and ‘Huang’ means yellow color , and is mainly used to treat sore throat.In China, S. baicalensis could be helpful for researchers, manufacturers, and policymakers to obtain a holistic view of this important herbal medicine.Despite the popular clinical use of Huang-Qin, scientific evidences are not adequate to identify the effective chemical components responsible for the versatile biological activities. The quality control of Huang-Qin crude drugs and related patent drugs still needs to be improved, and the medicinal potential of many bioactive compounds of this plant has yet to be explored. A comprehensive review of Scutellaria genus or S. baicalensis , and CNKI (Chinese). Scutellaria baicalensis, chemical constituents, phytochemistry, biological activities, and biosynthesis were used as the key words.Several review articles are available on the 1–126) and 6 polysaccharides have been isolated from S. baicalensis Georgi , 2 flavonols (43–44), 9 flavanones (45–53), 1 flavonol (54), 1 chalcone (55), and 1 biflavonoid (56). The most abundant ones are baicalein (1), wogonin (27), and oroxylin A (5). Wogonin is the first free flavonoid isolated from S. baicalensis, and its structure was established in 1930  is the most abundant compound of S. baicalensis. As the first pure compound reported from this plant, baicalin was originally reported by G. Bargellini in 1919 , O-glucuronides (73–89), and C-glycosides (90–100).Baicalin , kaempferol 3-O-β-d-glucoside (67), and 72 are exceptions. Compound 72 is an acylated anthocyanin containing two glucosyl residues at C-3 and C-5, and contributes to the blue (or purple) color of the flowers  and wogonoside (76) are the most abundant ones. For majority of these compounds, the glucuronyl group is linked to 7-OH, except for 79 (8-OH) and 80 (2′-OH).While glucuronides are not as prevalent as glucosides in plant secondary metabolites, C-glycosides were reported from S. baicalensis in 1994 . Their structures were mainly determined by NMR spectroscopic analysis. Unlike O-glycosides, the sugar residues are not easily hydrolyzed to identify their forms and stereo-configurations. Structures for some C-glycosides need to be further confirmed.The first two S. baicalensis. The aglycones are usually conjugated with a glucosyl group, which are further substituted with a rhamnosyl residue (Rha), or acylated with a caffeoyl (Caf) or feruloyl (Fer) group.A total of nine phenylethanoid glycosides have been reported from S. baicalensis include three steroids (109–111), one diterpene (112), five amides (113-117), and nine phenolic compounds (118–126). The amides are conjugates of isobutyl amine and organic acids, and were isolated from a water extract by Xu et al. -glucopyranose units, 8.3% of which are substituted with an α-glucopyranose unit at C-6 , and tentatively characterized 132 compounds in Huang-Qin by ultra-high performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry analysis (UHPLC/Orbitrap-MS)  has been widely used to characterize the chemical constituents in herbal extracts. A number of reports are available on chemical analysis of S. baicalensis within 20 min  and its major compounds such as baicalin, baicalein, and wogonin. These results were reported by different research groups, the investigations were conducted using different experimental models, and thus the results were difficult to be compared or summarized. Recently, our group isolated 28 compounds from this herb, and evaluated their anti-H1N1 viral, cytotoxic, and Nrf2 activation activities  murine model, oral administration of a water extract  led to 66% reduction of xenograft tumor  production via suppression of COX-2 (cyclooxygenase-2) expression and arachidonic acid release from cell membranes. The total free flavonoid extract  could also significantly reduce tumor size by 25.5% in A549 human lung cancer xenografted mice, via induction of growth arrest in S phase and inhibition of DNA synthesis , baicalein (1), wogonin (27), and wogonoside (76) are the major bioactive constituents responsible for the anti-tumor activities, with IC50 values of 10–50 μM against most tested cancer cell lines in vitro  could inhibit MDA468 breast cancer xenografts by 40%, the effect of which was comparable to that of the positive drug cisplatin (5 mg/kg). It could upregulate DDIT4 (DNA-damage-inducible transcript 4) expression, which mediated the inhibition of mTOR   inhibitors to potentiate the activities of anti-tumor drugs, such as the Bcl-2 (B-cell lymphoma 2) family inhibitor ABT-263. The combination of wogonin  and ABT-263 remarkably promoted tumor regression in human T-cell leukemia xenografted mice, but wogonin did not exhibit significant effects when used alone  could inhibit tumor growth of T47D and MDAMB-231 breast cancer xenografts by up to 88% without significant toxicity in athymic nude mice  , DENV on Vero cells (86.59–95.19 μg/mL), and H1N1 and seasonal influenza A viruses on MDCK cells (14.16–41.49 μg/mL)  showed significant effects in preventing death, prolonging survival time, inhibiting lung consolidation, and reducing the viral titers in the lung in BALB/c mice infected with the influenza A/FM1/1/47 (H1N1) virus. The effects were comparable to lamivudine. The mechanism could be inhibition of neuraminidase activity and modulation of the immune system  could reduce plasma duck hepatitis B virus (DHBV) DNA level in the liver of DHBV-infected ducks with an ED50 of 5 mg/kg, via inhibition of DHBV DNA polymerase and thus reducing the relaxed circular and linear forms of DHBV DNA  could remarkably inhibit influenza virus A/PR/8/34 (APR8) by reducing the replication of APR8 in MDCK cells, through inhibition of the fusion of the virus with endosome/lysosome membrane at early stage and inhibition of the budding of the progeny virus from the cell surface  in patients aged >1 year, rapidly relieving fever, attenuating oral lesions and rashes, and improving nervous system involvement. This result was derived from a multi-center and retrospective analysis , Moloney murine leukemia virus, and Rous-associated virus type 2 , including Streptococcus sanguis II, S. salivarius, Actinomyces viscosus, A. naeslundii, A. odontolyticus, two strains of Capnocytophaga, Bacteroides melaninogenicus ss intermedius, B. gingivalis, Fusobacterium nucleatum, and Actinobacillus actinomycetemcomitans  could protect mice from staphylococcal pneumonia caused by 43, 5,7,2′,6′-tetrahydroxyflavanonol, 40 mg/kg for one time, i.v.) could protect mice against a lethal challenge with heat-killed Escherichia coli 35218, increasing the survival rate from 0% to 60% via neutralization of LPS (lipopolysaccharide) and reduction of proinflammatory cytokines  showed potent anti-inflammatory activities in the zymosan-induced mice air-pouch, reducing NO production from 30 to 5 μM, through the down-regulation of IKKαβ (IκB kinase αβ) and NF-κB activation via suppression of c-Raf-1/MEK1/2 and MAPK phosphorylation  showed anti-inflammatory effects in double-stranded RNA (dsRNA)-induced macrophages by inhibiting NO, cytokines, chemokines, and growth factors via the endoplasmic reticulum stress-CHOP/STAT pathway  could protect cardiomyocytes in vitro from moderate hypoxia, ischemia/reperfusion, and antimycin A exposure, decreasing cell death from 47–49% to 23–26% by scavenging ROS (reactive oxygen species)   could improve rat act in the Morris water assay, reducing search error to about 50% in the chronic cerebral hypoperfusion and the LPS infusion models , and H2O2 induced toxicity in vitro  could significantly promote mouse hippocampal HT22 cell survival by 50% after injury induced by iodoacetic acid , CCl4, and LPS-induced rat or mouse hepatotoxity, by inhibiting cytokine, COX-2, iNOS (nitric oxide synthases), and NF-κB  and AST (aspartate transaminase) levels from 226 to 110 U/l and from 607 to 197 U/l, respectively  levels  pathway, and the PI3K/AKT signaling  could protect against the hyperglycemia-induced cardiovascular malformation during chick embryo development, decreasing the high incidence of cardiac bifida from 32% to 16%, by reducing ROS production and regulating SOD (superoxide dismutase), GSH-Px (glutathione peroxidase), and GABABaicalein could promote new blood vessel formation, attenuate cardiac remodeling and endothelium dysfunction against angiotensin II or myocardial ischemia reperfusion injury, via inhibition of AKT/mTOR, ERK1/2, NF-κB, and calcineurin sgnaling pathways in mice or chicks  could significantly increase bone mineral density by 12–18%, and improve bone trabecula microstructure of weightlessness induced osteoporosis rats via the osteogenic differentiation enhancement effect  could reduce physical symptoms associated with knee osteoarthritis in patients after 500 mg/d treatment for 1 week.The extract of S. baicalensis and baicalein  could reduce gastrointestinal dysfunction in ritonavir-treated rats  to form cinnamic acid. Interestingly, the subsequent biosynthetic steps were different for flavones in the aerial parts and in the roots (p-coumaroyl CoA ligase (4CL), chalcone synthase (CHS), chalcone isomerase (CHI), and flavone synthase (FNSII-1) to form apigenin  may participate in the biosynthesis of wogonin, though no OMT has been reported yet. Among the biosynthetic enzymes, SbCLL-7, SbCHS-2, FNSII-2 and F8H are expressed preferentially in the roots. Functions of these genes have been validated by RNAi in hairy roots of S. baicalensis and overexpression in transgenic Arabidopsis.The flavonoids in he roots . For theO-glucuronides and O-glucosides of S. baicalensis. SbUBGAT showed O-glucuronyltransferase activities for various flavones, and may take part in the biosynthesis of glucuronides like baicalin and wogonoside  have been extensively studied. Recently, our group reported the comprehensive correlations of chemicals and bioactivities of another popular herbal medicine Gan-Cao , and discovered a number of promising bioactive natural products (Ji et al. S. baicalensis, many post-modification enzymes have yet to be characterized, including those responsible for the hydroxylation, methylation, and glycosylation reactions.


The proteins IFITM1, IFITM2, and IFITM3 are host effectors against a broad range of RNA viruses whose roles in classical swine fever virus (CSFV) infection had not yet been reported. We investigated the effect of these proteins on CSFV replication in mammalian cells. The proteins were overexpressed and silenced using lentiviruses. Confocal microscopy was used to determine the distribution of these proteins in the cells, and immunofluorescence colocalization analysis was used to evaluate the relationship between IFITMs and the CSFV endosomal pathway, including early endosomes, late endosomes, and lysosomes. IFITM1, IFITM2, or IFITM3 overexpression significantly inhibited CSFV replication, whereas protein knockdown enhanced CSFV replication. In porcine alveolar macrophages (PAMs), IFITM1 was mainly located at the cell surface, whereas IFITM2 and IFITM3 were mainly located in the cytoplasm. Following CSFV infection, the distribution of IFITM1 changed. IFITM1, IFITM2, and IFITM3 colocalization with Lamp1, IFITM2 with Rab5 and Rab7, and IFITM3 with Rab7 were observed in CSFV-infected cells. Collectively, these results provide insights into the possible mechanisms associated with the anti-CSFV action of the IFITM family. Flaviviridae family and the Pestivirus genus [Classical swine fever virus (CSFV) causes disease in domestic pigs that affects the livestock industry with serious socioeconomic implications. ,2. CSFV us genus . CSFV isus genus ,5. Interus genus  and induus genus . Althougus genus ,10,11,12IFN-inducible transmembrane proteins (IFITMs) have been shown to have antiviral effects in viral infections, especially of enveloped viruses such as Ebola virus, influenza A virus, West Nile virus, hepatitis C virus, and dengue virus ,15,16,17IFITM1 interacts with CD81 and occludin, which are co-receptors of HCV, to disrupt HCV entry . IFITM2 In light of these data and given that IFITMs act as inhibitors in viral infection, we aimed to determine the effect of IFITMs on CSFV replication. In addition, our goal in the current work was to examine the distribution of IFITMs in PAMs and whether IFITMs affected CSFV infection.Human embryonic kidney cells  were cultured in Dulbecco’s minimal essential medium  supplemented with 10% fetal bovine serum . Porcine alveolar macrophages  were cultured in RPMI 1640 medium (Gibco) supplemented with 10% FBS. The Shimen strain of CSFV was obtained from the Control Institute of Veterinary Bio-products and Pharmaceuticals . Experiments involving CSFV were standardized according to the Laboratory Biosafety Manual and strictly performed in the P3 biosafety laboratory.IFITM1, IFITM2, IFITM3, and Mx1 , specific primers were designed  to generate CMV-IFITMs and cloned into pEGFP-C1 to generate C-IFITMs, a vector that expressed the IFITM fusion protein and enhanced green fluorescent protein (EGFP). One pair of short hairpin RNA (shRNAs) directed at IFITM1, IFITM2, and IFITM3 and the scrambled shRNA lentivirus (shN) control was designed with the RNAi Designer website (http://rnaidesigner.thermofisher.com/). shRNA was inserted into pCDH-U6-GreenPuro (pCDH-U6). The vectors were transfected into PAMs using Turbofect  according to the manufacturer’s instructions.g. Lentivirus titers were estimated in HEK-293T cells. Each of the lentiviruses was inoculated into 10 groups of cells in 96-well plate with a 10-fold dilution series (10−1–10−10), with the CMV vector or shN used as a negative control. The infected cells were detected with a fluorescence inversion microscope , and positive cells manifested by green fluorescence were counted after culture in DMEM with 2% FBS for 48 h. Lentivirus titers were expressed as 50% tissue culture infectious dose (TCID50) mL−1.CMV vector containing IFITMs  or pCDH-U6 vectors carrying IFITM shRNA lentiviruses (shIFITM) were serially co-transfected into HEK-293T cells using three ancillary plasmids, pGAG, pREV, and pVSV-G, by Turbofect. DMEM with 2% FBS was used to culture the transfected cells for 16 h, and then the medium was replaced with DMEM containing 10% FBS, 4.0 mM L-glutamine , 0.01 mM cholesterol , 0.01 mM L-α-phosphatidylcholine (Sigma), and 1:1000 diluted Chemically Defined Lipid (Invitrogen) followed by another 48 h of incubation. Three types of lentivirus  in the cell supernatant were collected by centrifugation at 1500× Four lentiviruses (CMV-IFITMs and shIFITMs), for which titers were sufficiently high for inoculation of PAMs with polybrene , were added to the culture medium to enhance the infection rate. The medium was refreshed at 8 h after infection of lentivirus, followed by another 48 h of incubation. Positive cells were selected with puromycin (Thermo Fisher Scientific). Empty CMV and shN served as negative controls. To survey the positive cells that exhibited green fluorescence, a fluorescence inversion microscope (Nikon) was used. The success of overexpression and knockdown of IFITMs was determined by western blot.Cell samples were mixed with radioimmunoprecipitation assay lysis and extraction buffer and lysed on ice. A small portion of the sample was used for quantitative determination using the BCA Protein Assay Kit . Residual protein samples were used for electrophoresis with 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene difluoride membranes . Following blocking in 5% non-fat milk, the membranes were incubated with antibodies including mouse anti-Flag monoclonal antibody , rabbit anti-IFITM1 polyclonal antibody , rabbit anti-IFITM2/3 PAb (Cusabio), mouse anti-GFP MAb , and mouse anti-β-actin MAb (CWBIO) at 4 °C overnight. After washing, peroxidase-conjugated goat anti-mouse (or rabbit) IgG secondary antibodies (CWBIO) were incubated with the membrane at room temperature for 2 h. Using chemiluminescence buffer, the protein bands were exposed.According to the manufacturer’s instructions, cell viability assay in IFITM-overexpression and -knockdown cell lines was performed using Cell Counting Kit-8 (CCK-8) .−1–10−10) and incubated for 24 h or 48 h. The negative control was generated by culture with medium without CSFV. All samples were fixed by 1:1 organic fixing liquid (acetone: methanol) at –20 °C for 20 min and permeabilized by 0.1% Triton X-100 at 4 °C for 20 min. After blocking with 5% skimmed milk, permeabilized cells were probed with positive serum against CSFV. Rabbit anti-pig IgG-fluorescein isothiocyanate (FITC) antibody (Sigma) was used following three washes with phosphate-buffered saline (PBS). Immunofluorescence was detected using a fluorescence inversion microscope (Nikon). Negative controls for background staining levels were prepared with mock cells. The viral titers were expressed as TCID50 mL−1.The virus titers of CSFV in the cellular supernatant were estimated by IFA. PAMs were inoculated with the supernatant of infected cells in a 96-well plate. Ten groups were prepared with a ten-fold dilution series , anti-Rab7 PAb (Santa Cruz Biotechnology), and anti-Lamp1 PAb , respectively, and probed using an Alexa Fluor 594 AffiniPure goat anti-rabbit IgG (H+L) antibody . The cells were stained with DAPI (Beyotime) at 25 °C for 10 min and imaged by laser scanning confocal microscopy .The cells were transfected with C1-IFITM1, -IFITM2, and -IFITM3 in 35-mmt-test was used for all statistical analyses. Data are presented as the mean ± standard deviation (SD) of three independent experiments. p-values < 0.05 were considered to indicate statistical significance.Student’s To analyze the effect of IFITM overexpression on CSFV replication, PAMs that stably expressed IFITM1, -2, or -3 were generated by a lentiviral packaging system. PAMs were transfected with empty plasmid (CMV) as a negative control. The cell viability assay suggested that the growth and viability of CMV-IFITM1, CMV-IFITM2, CMV-IFITM3, and CMV cells were similar to those of PAMs (approximately 100%) A. Green IFITM1, IFITM2, and IFITM3. PAMs were transfected with shIFITM or shN. A cell viability assay was performed in the cell lines. The results showed that the growth and viability of shN and shIFITM cells were similar to those of the control , 18 hpt, and 24 hpt  for 24 h. The mRNA and protein levels of IFITM1 were detected by RT-qPCR and western blot, respectively. The results showed that IFITM1 expression was significantly inhibited by CSFV infection at MOIs of 1 and 3 A,C. A siAfter we demonstrated that CSFV inhibits IFITM expression, we examined whether CSFV affected IFITM distribution. Because cellular localization of IFITM1, IFITM2, and IFITM3 had not been reported in PAMs, we constructed three plasmids  with IFITM1, IFITM2, and IFITM3 fused to EGFP to facilitate detection, respectively. Protein expression of IFITM1, IFITM2, and IFITM3 was confirmed by western blot A. PAMs tTo determine the mechanism of anti-CSFV activity by IFITMs, we examined whether CSFV attachment and entry could be obstructed by overexpression of IFITMs in PAMs, with regard to the successive steps of CSFV infection. To analyze viral binding, PAMs were incubated with CSFV at 4 °C to achieve viral attachment but impair viral entry. The excess virus was then washed away. CSFV genomic RNA collected at this point represented CSFV bound to the cell surface. As shown in It has been reported that CSFV entry into PK-15 cells and PAMs depends on endocytosis. In addition, both Rab5 and Rab7 are required for endocytosis and subsequent productive CSFV infection ,32. GiveMost ISGs promote innate immunity against viral infection and contribute to the antiviral activity of IFNs ,34. AmonIFITM1, IFITM2, and IFITM3, and the efficiency was analyzed in conditions of IFITM overexpression. We determined that overexpression of IFITM1, IFITM2, and IFITM3 in PAMs inhibited replication of CSFV at the mRNA level, reducing the viral titer. In contrast, CSFV replication was enhanced in lentivirus-mediated stable knockdown of IFITMs.Previous research of the swine IFITM family revealed that IFITM1, IFITM2, and IFITM3 share high amino acid homology ,22. Thusrns interacts with membrane-associated heparin sulfate or laminin receptor to mediate CSFV attachment [Although the mechanism of IFITM antiviral activity remains unclear, we confirmed that all three IFITMs influence CSFV entry. These findings are consistent with the inhibition of entry of vesicular stomatitis virus, West Nile virus, and dengue virus by IFITMs ,36. Envetachment ,38. Surptachment . Howevertachment . Rab5 antachment ,40,41,42tachment ,32. Rab5tachment . In thisIn viral infections, such as the dengue virus or influenza A virus, IFITM1 at the cell surface or in the early endosomal pathway suppresses virus entry, whereas IFITM3 prevents virus entry in the late endosomal pathway ,45. FurtCharacterization of the distribution of IFITM1, IFITM2, and IFITM3 in PAMs and localization patterns in the endosomal pathway upon overexpression were partly coincident with previous research in human cells, which revealed that IFITM1 is located at the cell surface and in early endosomes, while both IFITM2 and IFITM3 are located in late endosomes and lysosomes ,25. In tTaken together, we illustrated an important role for IFITMs in the inhibition of CSFV infection and revealed a link between the IFITM protein family and CSFV endosomal pathway. We postulated that IFITMs localize to distinct membranes and likely modify the membrane structure or alter endosomal physiology to impair viral membrane fusion. This will be the subject of our future studies.The exact mechanisms of anti-CSFV action by IFITM1, IFITM2, and IFITM3 warrant further elucidation. However, the evidence presented here provides insight into the close relationship between IFITM anti-CSFV action and the endosomal pathway, which is necessary for CSFV replication. Thus, IFITM1, IFITM2, and IFITM3 are considered potential anti-CSFV effectors that protect the host. This study improved our understanding of the antiviral action of IFITMs in CSFV infection and may contribute to the development of new anti-CSFV treatments.


In spite of significant advancements and success in antiretroviral therapies directed against HIV infection, there is no cure for HIV, which scan persist in a human body in its latent form and become reactivated under favorable conditions. Therefore, novel antiretroviral drugs with different modes of actions are still a major focus for researchers. In particular, novel lead structures are being sought from natural sources. So far, a number of compounds from marine organisms have been identified as promising therapeutics for HIV infection. Therefore, in this paper, we provide an overview of marine natural products that were first identified in the period between 2013 and 2018 that could be potentially used, or further optimized, as novel antiretroviral agents. This pipeline includes the systematization of antiretroviral activities for several categories of marine structures including chitosan and its derivatives, sulfated polysaccharides, lectins, bromotyrosine derivatives, peptides, alkaloids, diterpenes, phlorotannins, and xanthones as well as adjuvants to the HAART therapy such as fish oil. We critically discuss the structures and activities of the most promising new marine anti-HIV compounds. HIV targets immune cells and impairs the human defense against pneumonia, tuberculosis, and shingles as well as certain types of cancer -1,6-naphthyridine skeleton, namely 3-(phenetylamino)demethyl(oxy)aaptamine  and in peripheral blood mononuclear cells with EC50 of 0.032 µM. It was concluded that HIV-1 LTR NF-κB response elements are required for a bengamide A-mediated inhibition of LTR-dependent gene expression [Bengamide A 31, , haliclopression .34, Stachybotrys chartarum MXH-X73, was discovered to be a HIV-1 RT inhibitor, which showed inhibitory effects on the wild type (EC50 8.4 µM) and five NNRTI-resistant HIV-1 strains  [Phenylspirodrimane, stachybotrin D 34,  isolated 6.2 µM) .Dictyota pfaffii from Northeast Brazil: 10,18-diacetoxy-7-hydroxy-2,8(17)-dolabelladiene, 10,18-diacetoxy-7-hydroxy-2,8(17)-dolabelladiene, 10,18-diacetoxy-7-hydroxy-2,8-dolabelladiene, named dolabelladienols A–C  without a cytotoxic effect . Additionally, they investigated the virucidal effect of these diterpenes and their potential use as microbicides. Dolastane-diterpenes 38 and 40 showed a potent effect on HIV-1 infectivity, whereas no virucidal effect was observed for secodolastane diterpene 39, demonstrating another mechanism of antiretroviral activity. Therefore, the authors suggested a potential use of marine dolastanes 38 and 40 as microbicides that could directly inhibit virus infectivity and possibly act before the virus penetrates the target cells [De Souza Barros et al. tested marine dolastanes 38, 40,  and secoactivity . They obet cells .Dictyota spp has also been evaluated as a potential microbicide against HIV-1 in tissue explants. Namely, Stephens et al. examined the 8,10,18-trihydroxy-2,6-dolabelladiene ,11(S)-dolabella-3(E),7(E),12(18)-triene from Caribbean octocoral Eunicea laciniata [42–44, Although dolabellane diterpenes of brown alga aciniata . OxygenaEcklonia cava has shown an enhanced HIV-1 inhibitory effect [45, ′′′-dieckol inhibited the cytopathic effects of HIV-1, including HIV-1 induced syncytia formation, lytic effects, and viral p24 antigen production. Furthermore, 8,4′′′-dieckol inhibited an HIV-1 entry and RT enzyme with the inhibition ratio of 91% at a concentration of 50 μM.Phlorotannins are tannin derivatives made from several phloroglucinol units linked to each other in different ways. Phlorotannins contain phenyl linkage (fucols), ether linkage , phenyl and ether linkage (fucophloroethols), and dibenzodioxin linkage (eckols) ,107. So y effect ,113. Kar46, Aspergillus fumigates with fourteen other natural products [50 of 0.36 and 0.26 μM, respectively. A molecular docking study confirmed that PXA might bind to either CCR5 or CXCR4 to prevent HIV entry into target cells. Therefore, PXA, as a CCR5/CXCR4 dual-coreceptor antagonist, may be seen as a new potential lead product type for the development of anti-HIV therapeutics.Recently, for the first time, xanthone dimer was identified as a potential anti-HIV-1 agent . Xanthonproducts . However47, 48, HAART therapy can cause severe side effects, e.g., insulin resistance, lipoatrophy, dyslipidemia, and abnormalities of fat distribution. Therefore, finding an adequate diet and supplementation to lower the negative effects of the HAART combination therapy is desirable . Fish oiRecently, He et al. analyzed the influence of DHA on the locomotor activity in ethanol-treated HIV-1 transgenic rats . The pre49, 50, 51, H. fuscoatra exhibited a 92–98% reactivation efficiency of the latent HIV-1 relative to SAHA  and EC50 of 9.1, 39.6 and 6.3 µM [Resorcyclic acid lactones, namely radicicol 49,  pochonind 6.3 µM . The reaTruncatella angustata named truncateols O-V [52 and 53, 50 values of 39.0 μM and 16.1 μM, respectively. These compounds could be considered as new anti-HIV lead compounds due to lower cytotoxicity (CC50 > 100 μM) in comparison with the positive control efavirenz (CC50 = 40.6 μM).A team of researchers led by Zhao isolated new isoprenylated cyclohexanols from the sponge-associated fungus eols O-V . In vitrMarine organisms have been acknowledged as a precious source of bioactive compounds that may provide novel anti-HIV structures or lead structures for structural optimization. A large amount of evidence from scientific research confirmed a high biological potential of these compounds to treat serious diseases, including infective ones. Some of the marine-derived bioactive compounds discovered much earlier have emerged with novel properties and potential applications after a decade or two. Isolation and structural elucidation of compounds from marine organisms is not an easy task and still carries challenges. Identification of all the compounds is a daunting task, especially with regards to complex structural motifs that may be present in a single marine extract. Taxonomic knowledge is still insufficient to enable unambiguous species classification that can result in the false prediction of chemical constituents and hamper structural analysis. Furthermore, a temporal lag between the discovery, chemical characterization, and associated pharmacological activities is quite common, and the majority of marine metabolites are usually tested for anticancer activity, whereas anti-HIV and other possible biological effects are neglected or mostly not performed due to a lack of funding. Targeted assays and in vivo analyses are similarly performed only for some of the potential candidates, while the translation into clinical trials remains very limited. Thus, the financial gap is certainly a relevant factor contributing to the slow drug development process in this area. In particular, the development of anti-HIV compounds, which act by mechanisms that differ from existing antivirals, requires a well-designed and focused approach to studying the mode of action. Libraries should be created for specifically defined crude extracts, their corresponding simplified fractions as well as for pure compounds for a well-balanced natural product discovery program. Additionally, there exist but few publications in which scientists have tried to modify known compounds of marine origin to improve their bioactivity. We are, however, continuously witnessing advancements in the deep-sea exploration technology, sampling strategies, genome sequencing, genome mining, genetic engineering, chemo-enzymatic synthesis, nanoscale NMR structure determination, and development and optimization of suitable fermentation strategies to ensure a continued supply of unique bioactive compounds from the oceans. Therefore, the grounds have been met for a broad, international effort based on scientific collaboration that would rely on well-equipped infrastructure and human resources as a prerequisite for a full advancement in the field and development of new drug candidates for the pharmaceutical market in the future.


Until now, more than 30 members of inositol phosphatases have been identified in plants, which are classified intofive families, including inositol polyphosphate 5-phosphatases (5PTases), suppressor of actin (SAC) phosphatases, SAL1 phosphatases, inositol monophosphatase (IMP), and phosphatase and tensin homologue deleted on chromosome 10 (PTEN)-related phosphatases. The current knowledge was revised here in relation to their substrates and function in response to abiotic stress. The potential mechanisms were also concluded with the focus on their activities of inositol phosphatases. The general working model might be that inositol phosphatases would degrade the InsP3 or phosphoinositides, subsequently resulting in altering Ca2+ release, abscisic acid (ABA) signaling, vesicle trafficking or other cellular processes.Inositol signaling is believed to play a crucial role in various aspects of plant growth and adaptation. As an important component in biosynthesis and degradation of  Myo-inositol  and its derivative metabolites are ubiquitous in all eukaryotes as both lipids and soluble compounds playing important roles in stress responses, development, and many other processes . DA. DA2 canP C (PLC) channel . DAG can channel . Those i tissues . InsP6 hPx-InsPs . NotablyArabidopsis thaliana. These plant inositol phosphatases have a broad function in development and adaptation by altering the IP or PI signaling pathways. The general information of those Arabidopsis proteins was listed in Among the processes of inositol phosphate (IP) and the phosphoinositide (PI) signaling pathway, dephosphorylation is catalyzed by specific inositol phosphatases on various substrates . Until nArabidopsis, 21 in rice, and 39 in soybean [3 and InsP4); group II the water-soluble inositol polyphosphates and the membrane-bound phosphoinositide; group III, InsP4 and PtdInsP3 with a 3-position phosphate group; and group IV only phosphoinositide. Similar as the mammalian counterparts, plant 5PTases also have various substrate specificities. The substrates have been identified by biochemical evidences for twelve of the fifteen Arabidopsis 5PTases, including Group I, Group II, and Group IV 5PTases P3 pathway and alter abscisic acid (ABA) signaling, Ca2+ release, and reactive oxygen species (ROS) production [The 5PTases family is the biggest family of the mentioned inositol phosphatases, containing 15 members in  soybean . 5PTases soybean . Group I 5PTases . Since soduction ,20,21.Arabidopsis [Arabidopsis SAC phosphatases have a ubiquitous expression pattern, except for AtSAC6 which is only expressed in flowers under normal growth condition. Their expression was not altered by treatment with phytohormones  [AtSAC6 has been identified to be induced by salt stress, indicating it would be involved in salt response [sac9 mutants exhibit a constitutive stress response with highly up-regulated stress-induced genes and over-accumulation of ROS [2, PtdInsP2, and PtdIns4P, in addition to having a possible role in vesicle trafficking [The SAC phosphatases are polyphosphoinositide phosphatases, containing the enzymatic SAC domain . There abidopsis . Most Arand ABA) . When twresponse . Besidesn of ROS . Though fficking ,25,26.Arabidopsis [AtPTEN2a and AtPTEN2b were up-regulated at transcriptional level, but not at protein level under salt and osmotic stress [Comparing to 5PTases and SAC phosphatases, there are fewer members in the SAL, IMP, and PTEN families and most of them behave as bifunctional enzymes . AtSAL1 bidopsis . IMP andbidopsis ,66. The bidopsis ,68,69, wbidopsis . The trac stress , suggest3 is believed to be vital in the termination of IP3, consequently altering Ca2+ oscillations, ABA signaling, and other stress-related pathways. The transgenic Arabidopsis plants overexpressing mammalian type I (group I) inositol polyphosphate 5-phosphatase (InsP 5-ptase) exhibited increased drought tolerance with less water loss [3 and IP6 were decreased in the transgenic lines as expected, thus attenuating ABA induction and Ca2+ signal transduction. The stomata were less responsive to the inhibition of opening by ABA and more sensitive to ABA-induced closure. Furthermore, the microarray data showed that dehydration-responsive element-binding protein 2A (DREB2A), encoding a drought-inducible ABA-independent transcription factor, and the DREB2A-regulated genes were induced in the InsP 5-ptase transgenic plants, suggesting the drought tolerance is mediated via the DREB2A-dependent pathway [The capacity of 5PTases hydrolyzing IPter loss . The con pathway .3 signaling, thus altering of ABA pathway and Ca2+ release, which is believed to be vital in stress tolerance [For plant 5PTases, it is common to take a role in the degradation process of inositol phosphate or phosphoinositide, terminating the IPolerance ,21,34,39olerance ,36,41.At5PTase7 or At5PTase9 increased salt sensitivity and the overexpression plants increased salt tolerance [At5PTase7 or At5PTase9 reduced ROS production in the Arabidopsis roots after 10 to 15 min after salt treatment. Additionally, the expression of salt-responsive genes, such as RD29A and RD22, was not induced highly in both mutants as in the wild-type under salt stress [At5PTase7 or At5PTase9 would disturb ROS production and salt-responsive gene expression, probably hampering the subsequent rescue signal transduction. Interestingly, the At5PTase9 mutants appeared to have a better ability to resistant osmotic stress. Meanwhile, the At5PTase9 mutants decreased Ca2+ influx and fluid-phase endocytosis [The T-DNA insertion mutants of olerance ,36. Mutat stress ,36. It sAt5TPase13 showed a reduction of root growth under limited nutrient conditions and germination rates in response to sugar stress, along with ABA insensitivity [At5ptase13 mutants accumulate less IP3 in response to sugar stress [2+ to regulate PHOYOTROPIN1 signaling under blue light [At5TPase13 is one of the four At5TPases (At5TPase12-15), which contain the plant specific WD40 repeats ,44. The sitivity . The yeasitivity . The genr stress . Again, 2+ signaling, ABA responses, ROS generation, vesicle trafficking, and possible connection with other phytohormones [Besides, several transcriptional analyses showed that the expression of multiple At5PTases is greatly up- or down-regulated in response to a series of abiotic stresses, such as cold, osmotic, salt, drought, oxidative, and heat ,36,71. Chormones ,71, it cArabidopsis and also well-known as FIERY1 (FRY1), has dual enzymatic activity of inositol phosphatase and nucleotidase, which play a role in both inositol signaling and nucleotide metabolite [3, thus affecting the subsequent steps in a similar pattern of 5PTases, which we have discussed above.AtSAL1, identified as a homologue of the yeast HAL2 in tabolite ,48. AtSAtabolite ,63,72,73AtSAL1 could increase lithium tolerance in yeast by modifying Na+ and Li+ effluxes [GmSAL1, could alleviate salinity stress in tobacco BY-2 cells [AtSAL1 would cause more sensitivity to salt, osmotic, and cold stress in Arabidopsis [Atsal1 mutant, hos2 with a single amino acid substitution exhibited as more resistant to lithium and salt stress [AtSAL1 or ectopic expression of GmSAL1 could not enhance salt tolerance in Arabidopsis [AtSAL1 would enhance drought and cadmium resistance in Arabidopsis, suggesting it would be a negative regulator of stress tolerance [SAL1, by inserting the META motif from black yeast Aureobasidium pullulans, ApHal2, improved salt and drought tolerance in Arabidopsis [It seems the effects of AtSAL1 on stress responses are controversial. Ectopic expression of effluxes . Ectopic-2 cells . Mutatiobidopsis ,72. Howebidopsis ,74. Lossolerance ,63. Exprbidopsis . It seem3 signaling. The Atsal1 mutants increase IP3 accumulation and the expression levels of ABA and stress genes, including RD29A, cold-specific CRT-binding factor 2 (CBF2), and CBF3 [GmSAL1 leads to a reduction of IP3 accumulation and suppression of the ABA-induced stomatal closure [2+ release and modulate the auxin pathway by IP3 signaling in plant development [3 signaling as for 5PTases. Maybe further investigation will supply evidence that AtSAL1 takes a role in Ca2+ release and its downstream signaling in response to abiotic stress as well. Secondly, AtSAL1 also regulates the ion homeostasis via the IP3 pathway. Ectopic expression of AtSAL1 could modify Na+ and Li+ effluxes in yeast for lithium and salt tolerance [GmSAL1-transgenic BY-2 cells could compartmentalize more Na+ in vacuolar for protection from salt stress [3, accumulated in the Atsal1 mutants, when suffering stresses [AtSAL1-deficient mutants have been found to attenuate endoplasmic reticulum (ER) stress under cadmium stress [The molecular mechanism of AtSAL1 in stress responses seems to be complicated for its multiple effects in various cellular processes. First, AtSAL1 would regulate stress tolerance and ABA responses via IPand CBF3 . On the  closure . Furtherelopment ,55. It solerance . GmSAL1-t stress . Additiostresses ,52,53,65stresses . AtSAL1 m stress . But no m stress .Arabidopsis have also been characterized as multi-functional enzymes involved in inositol, ascorbate, and histone biosynthesis [Oryza sativa L.), chickpea (Cicer arietinum L.), soybean (Glycine max), barley (Hordeum vulgare), and Medicago truncatula [Arabidopsis [Arabidopsis IMP on stress tolerance yet. Only some authors have tried assays in chickpea and rice suggesting that IMPs also function in response to abiotic stress [IMPs were first identified in tomato to play a role in inositol synthesis with high sensitivity to lithium . Their hynthesis ,59,66, suncatula ,69,75. Tbidopsis . Chickpebidopsis . But fewc stress ,68,69. BArabidopsis IMP and IMP activity is increased in chickpea seedlings under abiotic stresses, including salt, cold, heat, dehydration, and paraquat. It is consistent with the results of the transcript analyses by qRT-PCR, which showed that CaIMP is induced under abiotic stress and ABA treatment [CaIMP-transgenic Arabidopsis plants exhibited enhanced tolerance to abiotic stress, whereas the IMP-deficient Arabidopsis mutants increased the sensitivity to stress during seed germination and seedling growth. The inositol content and ascorbate content of the CaIMP-overexpressing lines are higher than the wild-type and the vector control, suggesting CaIMP would improve the plant tolerance to stress through both metabolic pathways [CaIMP [CaIMP are different between the drought-tolerant and drought-susceptible accessions, suggesting this variation might regulate phytic acid contents in plants, thus conferring drought tolerance in chickpea [Biochemical evidence demonstrated that CaIMP contains the same enzyme activity as reatment . The CaIpathways . Associas [CaIMP . The trachickpea . In anotchickpea .OsIMP is significantly upregulated by cold and ABA treatment by transcript analyses [OsIMP in tobacco improved cold tolerance. The transgenic plants contained more inositol content, less hydrogen peroxide (H2O2), and less malondialdehyde (MDA), with increased antioxidant enzyme activities under normal and cold stress conditions [OsIMP would modulate the antioxidant enzymes’ activities to conquer cold stress.Rice analyses . The pronditions . It sugg3. Components involved in the inositol pathways have been noted for their general roles in stress tolerance. This article focused on the knowledge about inositol phosphatases, which are considered to be more important in the degradation pathway of IP3 signaling, and their function in plant responses to abiotic stress.Substantial evidences demonstrate inositol phosphates, phosphoinositides, and the related inositol signaling play a crucial role in various life processes of development and environmental adaptation in plants ,4,6,7,123 or phosphoinositides, thus facilitating Ca2+ release from cellular stores and affecting ABA or other phytohormones’ pathways. For their effects on lipid-bound phosphoinositides, several enzymes have been proved to be involved in vesicle trafficking. For most of the inositol phosphatases, the existed evidences could only support part of the model. There are also some other puzzles. Since phytic acid (InsP6) could also serve as a signaling molecule to regulate Ca2+ release [Arabidopsis. Study from other plants is relatively rare. Do these inositol phosphatases take a universal role in all plants under abiotic stress? Hopefully, more exploration will expand our understanding about inositol phosphatases.Around 30 members of inositol phosphatases from five families have been identified. Their functions and mechanisms are still largely unknown. Biochemical and physiological data, especially those from analytical techniques, have delineated their substrates and the affecting signals. Moreover, the genetic evidences elucidate the genes’ function and how to pass the signals. In general, loss-in-function of inositol phosphatases usually cause the accumulation of IPacid InsP could al


Dalbergia cultrata, Eriosema chinense, Erythrina suberosa, Millettia pendula, Sesbania grandiflora, Tadehagi triquetrum, Andrographis echioides, Barleria cristata, Justicia gendarussa, Premna integrifolia, Vitex trifolia, Acacia pennata, Cassia auriculata, Croton oblongifolius and Glycomis pentaphylla. Investigation of the phytochemical constituents, biological and pharmacological activities of the selected medicinal plants is reported. This study aims at providing a collection of publications on the species of selected medicinal plants in Myanmar along with a critical review of the literature data. As a country, Myanmar appears to be a source of traditional drugs that have not yet been scientifically investigated. This review will be support for further investigations on the pharmacological activity of medicinal plant species in Myanmar.Medicinal plants are a reservoir of biologically active compounds with therapeutic properties that over time have been reported and used by diverse groups of people for treatment of various diseases. This review covers 15 selected medicinal plants distributed in Myanmar, including  Natural products, especially those derived from plants, have been used to help mankind sustain human health since the dawn of medicine. Traditional medicine has been in existence since time immemorial and has been well accepted and utilized by the people throughout history. Since ancient times, plants have been an exemplary source of medicines. Plant-derived medicinal products have attracted the attention of scientists around the world for many years due to their minimum side effects and positive effects on human health.In the pharmaceutical landscape, plants with a long history of use in ethnomedicine can be a rich source of substances for the treatment of various ailments and infectious diseases. Medicinal plants are considered a repository of numerous types of bioactive compounds possessing varied therapeutic properties. The vast array of therapeutic effects associated with medicinal plants includes anti-inflammatory, antiviral, antitumor, antimalarial, and analgesic properties.According to the World Health Organization (WHO), a variety of drugs are obtained from different medicinal plants and about 80% of the world’s developing population depends on traditional medicines for their primary health care needs . MyanmarWe previously reported  the tradA total of 15 medicinal plants, which belong to seven families and 15 genera were recorded in this review. The results are summarized in Until now, few sources of information are available for some medicinal plants in the literature, and the sources of information in the literature make evident that additional new investigations and scientific reports are necessary.D. cultrata, locally known as Yin-daik, is widely distributed in Myanmar , fl, flA. ec82) (83) , echioidide (86) ,58, and ide (87)  and a chby GC-MS .82), and 5,7,8-trimethoxyflavone (98) isolated from A. echioides are reported to possess potent anti-inflammatory activity, with IC50 values of 37.6 ± 1.2 μM and 39.1 ± 1.3 μM, respectively, and to suppress NO inhibition activity . Be. Be100),is plant . Biologiis plant ,75, and is plant ,77. TherJ. gendarussa (Family: Acanthaceae) is a shade-loving, quick-growing, evergreen plant mostly found in moist areas. In Myanmar, it is commonly known as pha-wa-net and distributed in Bago, Kachin, Mandalay, Shan, Taninthayi and Yangon. J. gendarussa is native to China and is distributed widely across India, in tropical and subtropical areas of Asia, Sri Lanka and Malaysia -8-C-β-glucopyranoside (160), together with isorhamnetin 3-O-α-l-rhamno-pyranoside (161), kaempferol 3-O-α-l-rhamnopyranosyl-(1→4)-β-d-glucopyranoside (162), and isovitexin (163) , (2S)-7,4-dihydroxyflavan (4β→8)-catechin (188) [inhibition %: 33.5 ± 2.9 (p < 0.01) at 30 μM], (2S)-7,4-dihydroxyflavan (4β→8)-gallocatechin (189) [inhibition %: 28.2 ± 4.7 (p < 0.01) at 30 μM], and (−)-epigallocatechin (192) [inhibition %: 24.4 ± 3.1 (p < 0.01) at 30 μM], isolated from C. auriculata displayed hepatoprotective effects against cytotoxicity induced by D-GalM in primary cultured mouse hepatocytes quinoline, skimmianine and arborinine , f, f219) a F (224) , carbazoborinine , three p228–230)  and two  B (232)  were iso species  and deritaphylla . In 1966taphylla  and follle (240)  from the0) [241) , glycozo1) [242)  from thelycolone , three faplopine  were isotaphylla .G. pentaphylla showed antimicrobial, antioxidant and cytotoxic effects [G. pentaphylla, showed significant inhibitory activity towards the growth of crown gall tumors produced by Agrobacterium tumefaciens in a potato disc bioassay [G. pentaphylla showed hepatoprotective activity against paracetamol-induced hepatotoxicity in Swiss albino mice [G. pentaphylla ethanol extract in reducing the proliferation of Hep3 B cells [227), glycosmisines A (231) and B (232) displayed significant the cytotoxic activity against human cancer A549, HepG-2 and Hun-7 cell lines, showing with an IC50 values of 56.05, 43.68 and 57.01 μM for A549 cell line, 60.06, 50.30 and 62.89 μM for HepG-2 cell line and 73.16, 30.60 and 62.87 μM for Huh-7 cell line, respectively [G. pentaphylla ethanolic crude extract showed a significant antioxidant activity [The methanolic extract of the leaves and stems of  effects . The acrbioassay . The metino mice . Streeji B cells . Biscarbectively . G. pentactivity . AccordiErythrina suberosa might have potential as a future anticancer therapeutic based on the literature. The plants, Croton oblongifolius, Eriosema chinense, Premna integrifolia and Erythrina suberosa, their phytochemical constituents and biological activities are extensively investigated. Studies on the plants of Acacia pennata, Cassia auriculata, Glycomis pentaphylla and Tadehagi triquetrum have reported the presence of several bioactive compounds whose biological and pharmacological applications can further be investigated. The chemical and pharmacological action of Vitex trifolia have mainly emphasized on fruits, however, the chemical constituents of other parts of this plant and some traditional uses are still missing. On the other hand, the biological activities of the various extracts of Sesbania grandiflora, and Andrographis echioides have been reported scientifically but there have been a few reports on chemical constituents. There are few phytochemistry and biological studies of Dalbergia cultrata and Millettia pendula, Barleria cristata and Justicia gendarussa; Acacia pennata and Cassia auriculata, therefore, they seem to be interesting for investigation of new pharmaceuticals.In this review, we have systematically summarized the structures and properties of 242 compounds isolated from 15 species of selected Myanmar medicinal plants. Among the 15 selected medicinal plants that have been described in this review, A vast reservoir of selected species remains untapped in terms of phytochemical constituents, as well as pharmacology, and this is the research gap for future studies. Further investigations of endemic plants in Myanmar and their phytochemical constituents are necessary to completely understand the molecular mechanisms of their action in vivo and in vitro and to assure the plant extracts are safe for human use.