Indian medicinal plants are effective in the treatment and management of COVID-19

Indian medicinal plants are referred to as the“nectar of life”owing to their phytochemicals and bioactive complexes that are beneficial in treating diseases.Coronavirus disease 2019(COVID-19)is a global health issue without any proper medication.The indigenous plants of India can be exploited to control the precise signs of SARS-CoV-2.The Ministry of AYUSH(Ayurveda,Yoga and Naturopathy,Unani,Siddha,and Homeopathy)has advised routine usage of medicinal plants for COVID-19.Medicinal plants like Zingiber officinalis,Azadirachta indica,Ocimum sanctum,Nigella sativa,Withania somnifera,Curcuma longa,Piper nigrum,Allium sativum,Tinospora cordifolia,etc.have immunity-boosting,antiviral,antibacterial,antioxidant and anti-inflammatory actions that can suppress and treat symptoms of COVID-19.In vitro,in vivo as well as in silico validation,these phytochemicals can help us to manage and treat COVID-19 disease.This integration of traditional knowledge in the prophylaxis of corona infection and current skills validating it for the development of precise and powerful therapeutic approaches will more efficiently resolve different clinical aspects of COVID-19.The review focuses on both traditional and emergent methods to prevent and treat COVID-19 with various Indian medicinal plants along with their phytochemicals.

Regulation of cytokine signaling pathways by PIAS proteins

Cytokines activate multiple signal transduction pathways to regulate gene expression. STATs and NF-kB are two important families of transcription factors activated by cytokines. Abnormal regulation of STAT and NF-kB activities has been associated with human diseases. The protein inhibitor of activated STAT （PIAS） protein family has been proposed to interact with over 60 proteins, many of which are transcription factors involved in the immune system. PIAS proteins regulate transcription through several mechanisms, including blocking the DNA-binding activity of transcription factors, recruiting transcriptional co-repressors and promoting protein sumoylation. This article is to review the role of PIAS proteins in the regulation of STAT and NF-kB signaling pathways.

Autophagy Plays a Potential Role in the Process of Sea Cucumber Body Wall “Melting” Induced by UV Irradiation

The changes of tissue appearances and structures in the process of UV-induced ＂melting＂ for sea cucumber （Stichopus japonicus） body wall were studied. And the localization and determination of acid phosphatase （ACP）, Cathepsin B and Cathepsin L activities were also investigated. The results show that the connective tissue was damaged with many hollows emerging and the regular collagen bundles were broken apart into irregular fragments. Margination of condensed chromatin at the nuclear membrane was observed. Both Golgi＇s body and endoplasmic reticulure swelled, curled, and eventually double- or multi-lamellar vesicles were formed. A number of autophagic vesicles distributed in all through the whole cytoplasm. ACP becomes more active after UV irradiation. The activities of cathepsin B and cathepsin L increased in UV-treated sea cucumbers and both achieved their maximum under certain conditions, It indicates that autophagy plays a potential role in the ＂melting＂ process for sea cucumber body wall induced by UV irradiation.

Roles of CREB-binding protein (CBP)/p300 in respiratory epithelium tumorigenesis

CREB-binding protein （CBP） and its homologue p300 are transcriptional co-activators of various sequence-specific transcription factors that are involved in a wide array of cellular activities, such as DNA repair, cell growth, differentia- tion and apoptosis. Several studies have suggested that CBP and p300 might be considered as tumour suppressors, with their prominent role being the cross-coupling of distinct gene expression patterns in response to various stimuli. They exert their actions mainly via acetylation of histones and other regulatory proteins （e.g. p53）. A major paradox in CBP/ p300 function is that they seem capable of contributing to various opposed cellular processes. Respiratory epithelium tumorigenesis represents a complex process of multi-step accumulations of a gamut of genetic and epigenetic aberrations. Transcription modulation through the alternate formation of activating and repressive complexes is the ultimate converging point of these derangements, and CBP/p300 represents key participants in this interplay. Thus, illumination of their molecular actions and interactions could reveal new potential targets for pharmacological interventions in respiratory epithelium carcinogenesis.

Role of adipose-derived stromal cells in pedicle skin flap survival in experimental animal models

The use of skin flaps in reconstructive surgery is the first-line surgical treatment for the reconstruction of skin defects and is essentially considered the starting point of plastic surgery. Despite their excellent usability, their application includes general surgical risks or possible complications, the primary and most common is necrosis of the flap. To improve flap survival, researchers have used different methods, including the use of adiposederived stem cells, with significant positive results. In our research we will report the use of adipose-derived stem cells in pedicle skin flap survival based on current literature on various experimental models in animals.

Upgraded role of autophagy in colorectal carcinomas

Autophagy is a basic catabolic process closely asso-ciated with degradation of cellular components. The role of autophagy in colorectal cancer(CRC) remains controversial. The mechanism of autophagy has been identified as protecting mechanism against tumorige-nesis by isolation of damaged organelles or as cytopro-tective provides energy in hypoxic regions of CRC tumors. Mutations in proto-oncogenes, such as RAS and BRAF, have been associated with autophagy initiation through signaling pathways of BRAF/MEK/ERK and PI3 K/AKT/m TOR. A combination therapy of chemotherapeutic agents and autophagy inhibitors such as hydroxychloroquine or immunotherapy might represent a major step that could be evaluated as a putative novel therapeutic strategy in CRC patients.

Hierarchical and selective roles of galectins in hepatocarcinogenesis, liver fibrosis and inflammation of hepatocellular carcinoma

Hepatocellular carcinoma(HCC)represents a global health problem.Infections with hepatitis B or C virus,non-alcoholic steatohepatitis disease,alcohol abuse,or dietary exposure to aflatoxin are the major risk factors to the development of this tumor.Regardless of the carcinogenic insult,HCC usually develops in a context of cirrhosis due to chronic inflammation and advanced fibrosis.Galectins are a family of evolutionarily-conserved proteins defined by at least one carbohydrate recognition domain with affinity forβ-galactosides and conserved sequence motifs.Here,we summarize the current literature implicating galectins in the pathogenesis of HCC.Expression of"proto-type"galectin-1,"chimera-type"galectin-3 and"tandem repeat-type"galectin-4 is up-regulated in HCC cells compared to their normal counterparts.On the other hand,the"tandemrepeat-type"lectins galectin-8 and galectin-9 are downregulated in tumor hepatocytes.The abnormal expression of these galectins correlates with tumor growth,HCC cell migration and invasion,tumor aggressiveness,metastasis,postoperative recurrence and poor prognosis.Moreover,these galectins have important roles in other pathological conditions of the liver,where chronic inflammation and/or fibrosis take place.Galectin-based therapies have been proposed to attenuate liver pathologies.Further functional studies are required to delineate the precise molecular mechanisms through which galectins contribute to HCC.

Antioxidant role of heme oxygenase-1 in prehepatic portal hypertensive rats

AIM： To study the effect of bilirubin on the oxidative liver status and the activity and expression of heine oxygenase-1 （HO-1） in rat liver injury induced by prehepatic portal hypertension. METHODS： Wistar male rats, weighing 200-250 g, were divided at random into two groups： one group with prehepatic portal hypertension （PH） induced by regulated prehepatic portal vein ligation （PPVL） and the other group corresponded to sham operated rats. Portal pressure, oxidative stress parameters, antioxidant enzymes, HO-1 activity and expression and hepatic sinusoidal vasodilatation were measured. RESULTS： In PPVL rats oxidative stress was evidenced by a marked increase in thiobarbituric acid reactive substances （TBARS） content and a decrease in reduced glutathione （GSH） levels. The activities of liver antioxidant enzymes, superoxide dismutase （SOD）, catalase （CAT） and glutathione peroxidase （GSH-Px） were also diminished while activity and expression of HO-1 were enhanced. Administration of bilirubin （5μmol/kg body weight） 24 h before the end of the experiment entirely prevented all these effects. Pretreatment with Sn-protoporphyrin IX （Sn-PPIX） （100 μg/kg body weight, i.p.）, a potent inhibitor of HO, completely abolished the oxidative stress and provoked a slight decrease in liver GSH levels as well as an increase in lipid peroxidation. Besides, carbon monoxide, another heme catabolic product, induced a significant increase in sinusoidal hepatic areas in PPVL group. Pretreatment of PPVL rats with Sn-PPIX totally prevented this effect CONCLUSION： These results suggest a beneficial role of HO-1 overexpression in prehepatic portal hypertensive rats.

Cross-regulation of the Nanog and Cdx2 promoters

The first cell fate choice in the mammalian embryo, the segregation of the inner cell mass （ICM） and trophectoderm （TE）, is regulated by the mutually antagonistic effects of the transcription factors, Oct4 and Cdx2, while the pluripotency factor, Nanog, is essential to specify the epiblast. We have analyzed the promoters of Nanog and Cdx2, and have found that these two transcription factors are likewise regulated reciprocally. Using an embryonic stem cell line with conditional TE differentiation, we show that Nanog overexpression suppresses the upregulation of TE markers, while Nanog knockdown upregulates the expression of TE markers. We further show that Nanog and Cdx2 bind to and repress each other＇s promoters. However, whereas Nanog knockout results in detectable Cdx2 expression in the ICM, we observe no overt disruption of blastocyst development, indicating that Nanog plays a subservient role to Oct4 in segregation of the ICM and TE.

Efficient differentiation of vascular smooth muscle cells from Wharton's Jelly mesenchymal stromal cells using human platelet lysate: A potential cell source for small blood vessel engineering

BACKGROUND The development of fully functional small diameter vascular grafts requires both a properly defined vessel conduit and tissue-specific cellular populations.Mesenchymal stromal cells(MSCs) derived from the Wharton's Jelly(WJ) tissue can be used as a source for obtaining vascular smooth muscle cells(VSMCs),while the human umbilical arteries(h UAs) can serve as a scaffold for blood vessel engineering.AIM To develop VSMCs from WJ-MSCs utilizing umbilical cord blood platelet lysate.METHODS WJ-MSCs were isolated and expanded until passage(P) 4. WJ-MSCs were properly defined according to the criteria of the International Society for Cell and Gene Therapy. Then, these cells were differentiated into VSMCs with the use of platelet lysate from umbilical cord blood in combination with ascorbic acid,followed by evaluation at the gene and protein levels. Specifically, gene expression profile analysis of VSMCs for ACTA2, MYH11, TGLN, MYOCD, SOX9,NANOG homeobox, OCT4 and GAPDH, was performed. In addition,immunofluorescence against ACTA2 and MYH11 in combination with DAPI staining was also performed in VSMCs. HUAs were decellularized and served as scaffolds for possible repopulation by VSMCs. Histological and biochemical analyses were performed in repopulated h UAs.RESULTS WJ-MSCs exhibited fibroblastic morphology, successfully differentiating into"osteocytes", "adipocytes" and "chondrocytes", and were characterized by positive expression(> 90%) of CD90, CD73 and CD105. In addition, WJ-MSCs were successfully differentiated into VSMCs with the proposed differentiation protocol. VSMCs successfully expressed ACTA2, MYH11, MYOCD, TGLN and SOX9. Immunofluorescence results indicated the expression of ACTA2 and MYH11 in VSMCs. In order to determine the functionality of VSMCs, h UAs were isolated and decellularized. Based on histological analysis, decellularized h UAs were free of any cellular or nuclear materials, while their extracellular matrix retained intact. Then, repopulation of decellularized h UAs with VSMCs was performed for 3 wk. Decellularized h UAs were repopulated efficiently by the VSMCs. Biochemical analysis revealed the increase of total hydroyproline and s GAG contents in repopulated h UAs with VSMCs. Specifically, total hydroxyproline and s GAG content after the 1 st, 2 nd and 3 rd wk was 71 ± 10, 74 ± 9 and 86 ± 8 μg hydroxyproline/mg of dry tissue weight and 2 ± 1, 3 ± 1 and 3 ± 1μg s GAG/mg of dry tissue weight, respectively. Statistically significant differences were observed between all study groups(P<0.05).CONCLUSION VSMCs were successfully obtained from WJ-MSCs with the proposed differentiation protocol. Furthermore, h UAs were efficiently repopulated by VSMCs. Differentiated VSMCs from WJ-MSCs could provide an alternative source of cells for vascular tissue engineering.

Functional Inferences of Environmental Coccolithovirus Biodiversity

The cosmopolitan calcifying alga Emiliania huxleyi is one of the most abundant bloom forming coccolithophore species in the oceans and plays an important role in global biogeochemical cycling. Coccolithoviruses are a major cause of coccolithophore bloom termination and have been studied in laboratory, mesocosm and open ocean studies. However, little is known about the dynamic interactions between the host and its viruses, and less is known about the natural diversity and role of functionally important genes within natural coccolithovirus communities. Here, we investigate the temporal and spatial distribution of coccolithoviruses by the use of molecular fingerprinting techniques PCR, DGGE and genomie sequencing. The natural biodiversity of the virus genes encoding the major capsid protein （MCP） and serine palmitoyltransferase （SPT） were analysed in samples obtained from the Atlantic Meridional Transect （AMT）, the North Sea and the L4 site in the Westem Channel Observatory. We discovered nine new coccolithovirus genotypes across the AMT and L4 site, with the majority of MCP sequences observed at the deep chlorophyll maximum layer of the sampled sites on the transect. We also found four new SPT gene variations in the North Sea and at L4. Their translated fragments and the full protein sequence of SPT from laboratory strains EhV-86 and EhV-99B 1 were modelled and revealed that the theoretical fold differs among strains. Variation identified in the structural distance between the two domains of the SPT protein may have an impact on the catalytic capabilities of its active site. In summary, the combined use of ＇standard＇ markers （i.e. MCP）, in combination with metabolically relevant markers （i.e. SPT） are useful in the study of the phylogeny and functional biodiversity of coccolithoviruses, and can provide an interesting intracellular insight into the evolution of these viruses and their ability to infect and replicate within their algal hosts.

Functional characterization of(E)-β-caryophyllene synthase from lima bean and its up-regulation by spider mites and alamethicin

（E）-β-Caryophyllene is a sesquiterpene compound widely distributed in plants and functions in plant defence. However, little is known about the sequence and function of （E）-β-caryophyllene synthase in lima bean （Phaseolus lunatus）. Here, we report a new full-length cDNA （PICAHS） encoding （E）-β-caryophyllene synthase, a possible key enzyme of plant defence. The cDNA of PICAHS contains an open reading frame of 1 761 bp, encoding a protein of 586 amino acids with a predicted mass of 67.95 kDa. The deduced amino acid sequence shows 52% identity with sesquiterpene synthase MtCAHS of Med- icago truncatula. Based on phylogenetic analysis, PICAHS is classified as the terpene synthases （TPS）-a subfamily. The recombinant enzyme, expressed in Escherichia coil, catalysed the formation of a major product （E）-β-caryophyllene （82%） and a minor product a-humulene （18%） from farnesyl dJphosphate. Real-time quantitative PCR （qRT-PCR） analysis found that the PICAHS transcript was significantly up-regulated in leaves after treatment with spider mites and alamethicin （ALA）, suggesting its ecological function in plant defence.

Role of autophagy in cholangiocarcinoma:An autophagy-based treatment strategy

Cholangiocarcinomas(CCAs)are diverse biliary epithelial tumours involving the intrahepatic,perihilar and distal parts of the biliary tree.The three entirely variable entities have distinct epidemiology,molecular characteristics,prognosis and strategy for clinical management.However,many cholangiocarcinoma tu-mor-cells appear to be resistant to current chemotherapeutic agents.The role of autophagy and the therapeutic value of autophagy-based therapy are largely unknown in CCA.The multistep nature of autophagy offers a plethora of re-gulation points,which are prone to be deregulated and cause different human diseases,including cancer.However,it offers multiple targetable points for designing novel therapeutic strategies.Tumor cells have evolved to use auto-phagy as an adaptive mechanism for survival under stressful conditions such as energy imbalance and hypoxic region of tumors within the tumor microenvir-onment,but also to increase invasiveness and resistance to chemotherapy.The purpose of this review is to summarize the current knowledge regarding the interplay between autophagy and cholangiocarcinogenesis,together with some preclinical studies with agents that modulate autophagy in order to induce tumor cell death.Altogether,a combinatorial strategy,which comprises the current anti-cancer agents and autophagy modulators,would represent a positive CCA patient approach.

Molecular basis of the first cell fate determination in mouse embryogenesis

Through proliferation and differentiation, a single cell, the zygote, can give rise to a complex organism composed of many types of cells. Up to the eight-cell embryo stage, the blastomeres are morphologically identical and distributed symmetrically in the mammalian embryo. Functionally, in some species, they are all totipotent. However, due to the compaction of blastomeres and the asymmetrical cell division at the late phase of the eight-cell embryo, the blastomeres of the morula are no longer identical. During the transition from morula to blastocyst, blastomeres differentiate, resulting in the first cell fate decision in embryogenesis, namely, the segregation of the inner cell mass and the tropheetoderm. In this review, we will discuss the regulatory mechanisms essential for the cell fate choice during blastocyst development, including transcriptional regulation, epigenetic regulation, mieroRNAs, and signal transduction.

Diversity and selection of the continuous-flowering gene, RoKSN, in rose

Blooming seasonality is an important trait in ornamental plants and was selected by humans.Wild roses flower only in spring whereas most cultivated modern roses can flower continuously.This trait is explained by a mutation of a floral repressor gene,RoKSN,a TFL1 homologue.In this work,we studied the origin,the diversity and the selection of the RoKSN gene.We analyzed 270 accessions,including wild and old cultivated Asian and European roses as well as modern roses.By sequencing the RoKSN gene,we proposed that the allele responsible for continuous-flowering,RoKSN copia,originated from Chinese wild roses(Indicae section),with a recent insertion of the copia element.Old cultivated Asian roses with the RoKSN copia allele were introduced in Europe,and the RoKSN copia allele was progressively selected during the 19th and 20th centuries,leading to continuous-flowering modern roses.Furthermore,we detected a new allele,RoKSN A181,leading to a weak reblooming.This allele encodes a functional floral repressor and is responsible for a moderate accumulation of RoKSN transcripts.A transient selection of this RoKSN A181 allele was observed during the 19th century.Our work highlights the selection of different alleles at the RoKSN locus for recurrent blooming in rose.

HPLC-DAD analysis and antifungal effect of Hyptis martiusii Benth(Lamiaceae) against Candida strains

Objective: To evaluate the anti Candida activity of Hyptis martiusii decoction and its major compound, caffeic acid alone or in the presence of fluconazole, as well as their cytotoxic effect. Methods: The decoction was characterized using high performance liquid chromatography coupled with diode array detector. For the antifungal activity, the minimum inhibitory concentration(MIC) and the potential effect of the decoction with the fluconazole were evaluated by microdilution method using 96-well microtiter trays. The osmotic fragility test was performed using erythrocytes under saline stress. All tests were performed in triplicate. Results: The chemical characterization of the decoction was performed by high performance liquid chromatography and revealed the presence of seven compounds, including caffeic acid as major constituent. The antifungal tests demonstrated that both decoction(DHm) and caffeic acid obtained from Hyptis martiusii presented MIC and MFC ≥4096 μg/mL against Candida albicans and Candida tropicalis strains. However, in the presence of fluconazole, DHm and caffeic acid presented IC_(50) of 2.60 and 2.53 μg/mL respectively, demonstrating significant synergistic effects against Candida strains. The modulator activity of DHm might be due to the presence of caffeic acid. Moreover, DHm and caffeic acid did not cause significant hemolytic effects, indicating that they present low cytotoxicity. Conclusions: These data indicate that DHm potentiates the activity of the fluconazole, without enhancement of the toxicity, encouraging further toxicological, pharmacological and phytochemical studies to provide consistent evidence of the potential of this plant to be used in drug development.

Implication of gut microbiome in immunotherapy for colorectal cancer

Colorectal cancer(CRC) constitutes the third most frequently reported malignancy in the male population and the second most common in women in the last two decades. Colon carcinogenesis is a complex, multifactorial event, resulting from genetic and epigenetic aberrations, the impact of environmental factors, as well as the disturbance of the gut microbial ecosystem. The relationship between the intestinal microbiome and carcinogenesis was relatively undervalued in the last decade. However, its remarkable effect on metabolic and immune functions on the host has been in the spotlight as of recent years. There is a strong relationship between gut microbiome dysbiosis, bowel pathogenicity and responsiveness to anti-cancer treatment;including immunotherapy. Modifications of bacteriome consistency are closely associated with the immunologic response to immunotherapeutic agents. This condition that implies the necessity of gut microbiome manipulation. Thus, creatingan optimal response for CRC patients to immunotherapeutic agents. In this paper, we will review the current literature observing how gut microbiota influence the response of immunotherapy on CRC patients.

Antiproliferative Activity of Acerogenin C, a Macrocyclicdiarylheptanoid, on PDGF-Induced Human Aortic Smooth Muscle Cells Proliferation

Platelet-derived growth factor (PDGF)-BB is one of the most potent factors in the development and progression of various vascular disorders, such as atherosclerosis and restenosis. PDGF is a major stimulant for vascular smooth muscle cells (VSMCs) proliferation via the mitogenesis signaling pathway. In the present study, we investigated the effect of acerogenin C, a macrocyclicdiarylhep-tanoid, on PDGF-BB-stimulated human aortic smooth muscle cells (HASMCs) proliferation. Acer-ogenin C significantly inhibited PDGF (20 ng/mL)-BB-induced [3H]-thymidine incorporation into DNA at concentrations of 0.1, 1 and 10 μM without any cytotoxicity. Acerogenin C also blocked PDGF-BB-stimulated phosphorylation of PLCγ1 and Akt but had no effect on extracellular signal-regulated kinase 1/2 (ERK1/2) and PDGF beta-receptor (Rβ) activation. In addition, acerogenin C (0.1 - 10 μM) induced cell-cycle arrest in the G1 phase, which was associated with the down-regulation of cyclins and the up-regulation of p27kip1. These results suggest that acerogenin C blocks PDGF-BB-stimulated HASMCs proliferation via G0/G1 arrest in association with the induction of p27kip1 and the suppression of PLCγ1 and phosphatidylinositol 3-kinase (PI3-K)/Akt signaling pathways. Furthermore, acerogenin C may be used for prevention and treatment of atherosclerosis during restenosis after coronary angioplasty.

Anti-inflammatory effects ofα-humulene andβ-caryophyllene on pterygium fibroblasts

AIM:To investigate the anti-inflammatory effects of the sesquiterpenesα-humulene andβ-caryophyllene on pterygium fibroblasts.METHODS:Primary cultures of pterygium fibroblasts were established.Third passage pterygium fibroblasts were exposed toα-humulene andβ-caryophyllene separately and together.The cell viability was assessed by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide(MTT)assay at 12,24,48,and 72h after exposure.The levels of the inflammatory cytokines interleukin(IL)-1β,IL-6,IL-8,tumor necrosis factor(TNF)-αand IL-10 in the conditioned culture medium were assessed by enzymelinked immunosorbent assay(ELISA)at 12,24 and 48h after exposure.Data were statistically analyzed using Friedman repeated measures analysis of variances on ranks.RESULTS:The 25μmol/Lβ-caryophyllene induced significant decrease in the IL-6 production by pterygium fibroblasts 48h after the exposure(P=0.041).The levels of IL-1β,IL-8,IL-10,and TNF-αwere very low and had no statistically significant variations af ter exposure toα-humulene,β-caryophyllene,or both compounds together.CONCLUSION:The exposure to 25μmol/L ofβ-caryophyllene significantly reduce the production of IL-6 by pterygium fibroblasts after 48h.This sesquiterpene may be a potential alternative adjuvant agent for the treatment of pterygium.