Preliminary Studies on the Tissue Culture of Cannabis sativa L.(Industrial hemp)

The internodes of the new cultivar Long-ma No.1 of Cannabis sativa L.（Industrial hemp） were used as explants for tissue culture. The paper studied the key factors of industrial hemp tissue culture, such as the physiological state of aseptic seedlings, the selection and concentration of plant growth regulators and so on.Hemp seed disinfection used 75% alcohol for 2 min and sterilized in 1‰ Hg Cl2 for 5min. The best combinations of plant growth regulators were 1.0 mg/L 6-BA and 0.5mg/L NAA for the induction of callus, and the best combinations of hormones were1.0 mg/L KT and 0.5 mg/L NAA for differentiation rate of adventitious bud.

Study on the Anti-inflammatory and Repairing Effects of Cannabis Sativa Leaf Extract by Epikutis Model Substitution

To explore the anti-inflammatory effect of Cannabis sativa leaf extracts and their repair to damaged skin,a 3D epidermal model(Epikutis®)was used.During the experiment,after epithelial cell surface was pretreated with SLS for 24 h,the anti-inflammatory effect and repair effect of Cannabis sativa leaf extract were observed by testing tissue viability and the representative pro-inflammatory factors such as IL-1α,TNF-αand IL-8.The tissue viability was detected by the MTT test.The results showed that the Cannabis sativa leaf extract effectively inhibited the secretion of inflammatory factors IL-1α,TNF-αand IL-8,and the tissue viability of the skin model was significantly improved after using the sample.The Cannabis sativa leaf extract has a repairing effect on skin barrier damage by inhibiting skin inflammatory reaction.

A Comparative Study on Hemp(Cannabis sativa)Essential Oil Extraction Using Traditional and Advanced Techniques

A comparative study of Cannabis sativa(Hemp)essential constituents obtained by using Supercritical Fluid Extraction(SCFE),Steam Distillation(SD)and Hydrodistillation(HD)is presented here.The optimized extraction temperatures were 130,110and 50℃for hydrodistillation,steam distillation and supercritical fluid extraction respectively.The essential oil of C.sativa was analyzed by using Gas chromatography mass spectrometry(GC-MS).A total of 33,30and 31components have been identified in HD,SD and SCFE respectively.Yield of essential oil using SCFE(0.039%)was more than HD(0.025%)and SD(0.035%)extraction respectively.The main component of sesquiterpenes obtained by hydrodistillation at 130℃with their percentages included caryophyllene(40.58%),trans-α-bergamotene(5.41%),humulene(10.97%),cis-β-farnesene(8.53%)and monoterpenes includedα-pinene(2.13%),d-limonene(6.46%),p-cymol(0.65%)and cineole(2.58%)respectively.The main component of sesquiterpenes obtained by SD steam distillation at110℃including caryophyllene(38.60%)trans-α-bergamotene(4.22%),humulene(10.26%),cis-β-farnesene(6.67%)and monoterpenes includedα-pinene(3.21%),d-limonene(7.07%),p-cymol(2.59%)and cineole(3.88%)whereas the more percentages of major components were obtained by SCFE at 50℃included caryophyllene(44.31%),trans-α-bergamotene(6.79%),humulene(11.97%)cis-β-farnesene(9.71%)and monoterpenes includedα-pinene(0.45%),d-limonene(2.13%)p-cymol(0.19%)and cineole(1.38%)respectively.We found yield/efficiency,chemical composition,quality of the essential oils by supercritical fluid extraction superior in terms of modern,green,saving energy and a rapid approach as compared to traditional techniques.

Modified Atmospheric Packaging and Its Effect on Postharvest Cannabis Quality

Cannabis sativa L. is used as fiber, food, and medicine in several countries. Though it is illegal for recreational use in most of the world, there are some countries that have legalized production and sale. There is a lot of research on production of cannabis, but less so on storage technologies. Cannabis contains several high value compounds, such as cannabinoids and terpenoids, that are susceptible to degradation via light, temperature, and oxygen. Several studies have explored temperature and light, and industry has adjusted accordingly. However, less is known about oxygen-induced degradation. Biochemical studies have demonstrated oxidative degradation of high value compounds, and many producers use some form of modified atmospheric packaging (MAP) for storage. However, the efficacy of MAP is unclear. The objective of this paper is to review our current understanding of MAP in postharvest cannabis storage and identify avenues where additional research is needed.

Endophytic fungi harbored in Cannabis sativa L.:diversity and potential as biocontrol agents against host plant-specific phytopathogens

The objective of the present work was isolation,phylogenetic characterization,and assessment of biocontrol potential of endophytic fungi harbored in various tissues(leaves,twigs,and apical and lateral buds)of the medicinal plant,Cannabis sativa L.A total of 30 different fungal endophytes were isolated from all the plant tissues which were authenticated by molecular identification based on rDNA ITS sequence analysis(ITS1,5.8S and ITS2 regions).The Menhinick’s index revealed that the buds were immensely rich in fungal species,and Camargo’s index showed the highest tissue-specific fungal dominance for the twigs.The most dominant species was Penicillium copticola that could be isolated from the twigs,leaves,and apical and lateral buds.A detailed calculation of Fisher’s log series index,Shannon diversity index,Simpson’s index,Simpson’s diversity index,and Margalef’s richness revealed moderate overall biodiversity of C.sativa endophytes distributed among its tissues.The fungal endophytes were challenged by two host phytopathogens,Botrytis cinerea and Trichothecium roseum,devising a dual culture antagonistic assay on five different media.We observed 11 distinct types of pathogen inhibition encompassing a variable degree of antagonism(%)on changing the media.This revealed the potential chemodiversity of the isolated fungal endophytes not only as promising resources of biocontrol agents against the known and emerging phytopathogens of Cannabis plants,but also as sustainable resources of biologically active and defensive secondary metabolites.

GC Determination of Active Cannabinoids in Cannabis of Chinese Origin and Survey of Their Contents

A method of GC determination using tetracosane as the internal standard for active cannabinoids cannabidiol.△~9-tetrahydrocannabinol and cannabinol of cannabis is described.The method is simple and accurate and has been applied to investigate the contents of active cannabinoids in cannabis of Chinese origin.Information was obtained from provinces and autonomous region of China. Three pure cannabinoids used as the reference substances were extracted and isolated successfully from cannabis.

Overcoming the Bell‐Shaped Dose‐Response of Cannabidiol by Using Cannabis Extract Enriched in Cannabidiol

Cannabidiol (CBD), a major constituent of Cannabis, has been shown to be a powerful anti-inflammatory and anti-anxiety drug, without exerting a psychotropic effect. However, when given either intraperitoneally or orally as a purified product, a bell-shaped dose-response was observed, which limits its clinical use. In the present study, we have studied in mice the anti-inflammatory and anti-nociceptive activities of standardized plant extracts derived from the Cannabis sativa L., clone 202, which is highly enriched in CBD and hardly contains any psychoactive ingredients. In stark contrast to purified CBD, the clone 202 extract, when given either intraperitoneally or orally, provided a clear correlation between the anti-inflammatory and anti-nociceptive responses and the dose, with increasing responses upon increasing doses, which makes this plant medicine ideal for clinical uses. The clone 202 extract reduced zymosan-induced paw swelling and pain in mice, and prevented TNFα production in vivo. It is likely that other components in the extract synergize with CBD to achieve the desired anti-inflammatory action that may contribute to overcoming the bell-shaped dose-response of purified CBD. We therefore propose that Cannabis clone 202 (Avidekel) extract is superior over CBD for the treatment of inflammatory conditions.

Preventive effects of cannabis on neurotoxic and hepatotoxic activities of malathion in rat

Objective: To investigate the effect of Cannabis sativa extract on the development of neuroand hepato-toxicity caused by malathion injection in rats. Methods: The extract of Cannabis sativa was obtained from the plant resin by chloroform treatment. Δ~9-Tetrahydrocannabinol content of the extract(20%) was quantified using gas chromatography–mass spectrometry. The doses of cannabis extract were expressed as Δ~9-tetrahydrocannabinol content of 10 or 20 mg/kg. Malathion(150 mg/kg) was intraperitoneally administered followed after 30 min by the cannabis extract(10 or 20 mg/kg, subcutaneously). Rats were euthanized 4 h later. Malondialdehyde(MDA), reduced glutathione(GSH), nitric oxide and paraoxonase-1(PON-1) activity were determined in brain and liver. Brain 5-lipoxygenase and butyrylcholinesterase(BChE) activity were measured as well. Histopathological examination of brain and liver tissue was also performed. Results: Compared to controls, malathion resulted in increased oxidative stress in brain and liver. MDA and nitric oxide concentrations were significantly increased(P<0.05) and GSH significantly decreased with respect to control levels(P<0.05). Malathion also significantly inhibited PON-1 and BChE activities but had no effect on brain 5-lipoxygenase. Brain MDA concentrations were not altered by cannabis treatment. Cannabis at 20 mg/kg, however, caused significant increase in nitric oxide and restored the GSH and PON-1 activity. Brain BChE activity significantly decreased by 26.1%(P<0.05) after treatment with 10 mg/kg cannabis. Cannabis showed no effect on brain 5-lipoxygenase. On the other hand, rats treated with cannabis exhibited significantly higher levels of liver MDA, nitric oxide and PON-1 activity compared with the malathion control group. Rats treated with only malathion exhibited spongiform changes, neuronal damage in the cerebral cortex and degeneration of some Purkinje cells in the cerebellum. There were also hepatic vacuolar degeneration and dilated and congested portal vein. These histopthological changes induced by malathion in brain and liver were reduced to great extent by cannabis administration at 20 mg/kg. Conclusions: Our data suggest that acute treatment with cannabis alleviates the malathion-induced brain and hepatic injury in rats possibly by maintaining the levels of GSH and PON-1 activity.

Acetylcholinesterase,butyrylcholinesterase and paraoxonase 1 activities in rats treated with cannabis,tramadol or both

Objective:To investigate the effect of Cannabis sativa resin and/or tramadol,two commonly drugs of abuse on acetylcholinesterase and butyrylcholinesterase activities as a possible cholinergic biomarkers of neurotoxicity induced by these agents.Methods:rats were treated with cannabis resin(5,10 or 20 mg/kg)(equivalent to the active constituent A'-tetrahydrocannabinol),tramadol(5,10 and 20 mg/kg) or tramadol(10 mg/kg)combined with cannabis resin(5,10 and 20 mg/kg) subcutaneously daily for 6 weeks.Acetylcholinesterase(AChE) and butyrylcholinesterase(BChE) activities were measured in brain and serum.We also measured the activity of paraoxonase-1(PONl) in serum of rats treated with these agents.Results:(i) AChE activity in brain increased after 10-20 mg/kg cannabis resin(by 16.3%-36.5%).AChE activity in brain did not change after treatment with 5-20 mg/kg tramadol.The administration of both cannabis resin(5,10 or 20 mg/kg) and tramadol(10 mg/kg) resulted in decreased brain AChE activity by 14.1%,12.9%and 13.6%,respectively;(ii) BChE activity in serum was markedly and dose-dependenlly inhibited by cannabis resin(by 60.9%-76.9%).BChE activity also decreased by 17.6%-36.5%by 10-20mg/kg tramadol and by 57.2%-63.9%by the cannabis resin/tramadol combined treatment;(iii)Cannabis resin at dose of 20 mg/kg increased serum PONl activity by 25.7%.In contrast,tramadol given at 5,10 and 20 mg/kg resulted in a dose-dependent decrease in serum PON1 activity by 19%,36.7%,and 46.1%,respectively.Meanwhile,treatment with cannabis resin plus tramadol resulted in 40.2%,35.8%,30.7%inhibition of PONl activity compared to the saline group.Conclusions:these data suggest that cannabis resin exerts different effects on AChE and BChE activities which could contribute to the memory problems and the decline in cognitive function in chronic users.

Market Status and Development of Cannabis Aativa Cosmetics in the Times of “Secondary-Constituent Community”

Since 2020,with the advent of the COVID-19 epidemic,the upsurge of the concept of“Constituent community”in the cosmetics industry has gradually subsided.The phenomenon of only focuing on the components but not on the synergies of the product system has gradually ebbed.On the other hand,with the popularity of the concept of industrial Cannabis sativa and Cannabis sativa cosmetics both at home and abroad,the Cannabis sativa cosmetics that replace the single effect propaganda of the Constituent community have been increasingly recognized by consumers,and industrial Cannabis sativa is increasingly widely used in cosmetics.For the development of Cannabis sativa cosmetics in the times of“Secondary-Constituent community”,the market status of Cannabis sativa cosmetics in China and abroad was analyzed in this paper.At the same time,the future development prospect of domestic Cannabis sativa cosmetics was preliminarily estimated and prospected,which was helpful for the development of Cannabis sativa cosmetics and provided the evaluation basis for the investment and development of domestic Cannabis sativa cosmetics.

Anticancer activity of Δ^(9)-tetrahydrocannabinol and cannabinol in vitro and in human lung cancer xenograft

Objective:To investigate the effects of Δ^(9)-tetrahydrocannabinol,the principal psychoactive compound of Cannabis sativa,and cannabinol,a Δ^(9)-tetrahydrocannabinol degradative product,on human non-small cell lung cancer cells.Methods:Δ^(9)-Tetrahydrocannabinol and cannabinol were tested for anticancer activity in human non-small cell lung cancer(A549)cells.The effects on cell proliferation,apoptosis,and phosphorylation profiles were examined.The effects of Δ^(9)-tetrahydrocannabinol and cannabinol on tumor growth were also investigated using a xenograft nude mouse model.Apoptosis and targeted phosphorylation were verified by immunohistochemistry.Results:Δ^(9)-Tetrahydrocannabinol and cannabinol significantly inhibited cell proliferation and increased the number of apoptotic cells in a concentration-dependent manner.The Δ^(9)-tetrahydrocannabinol-and cannabinol-treated cells had lower levels of phosphorylated protein kinase B[AKT(S473)],glycogen synthase kinase 3 alpha/beta,and endothelial nitric oxide synthase compared to the controls.The study of xenograft mice revealed that tumors treated with 15 mg/kg Δ^(9)-tetrahydrocannabinol or 40 mg/kg cannabinol were significantly smaller than those of the control mice.The tumor progression rates in mice treated with 15 mg/kg Δ^(9)-tetrahydrocannabinol or 40 mg/kg cannabinol were significantly slower than in the control group.Conclusions:These findings indicate that Δ^(9)-tetrahydrocannabinol and cannabinol inhibit lung cancer cell growth by inhibiting AKT and its signaling pathways,which include glycogen synthase kinase 3 alpha/beta and endothelial nitric oxide synthase.

Neuroprotective potential of cannabidiol:Molecular mechanisms and clinical implications

Cannabidiol(CBD),a nonpsychotropic phytocannabinoid that was once largely disregarded,is currently the subject of significant medicinal study.CBD is found in Cannabis sativa,and has a myriad of neuropharmacological impacts on the central nervous system,including the capacity to reduce neuroinflammation,protein misfolding and oxidative stress.On the other hand,it is well established that CBD generates its biological effects without exerting a large amount of intrinsic activity upon cannabinoid receptors.Because of this,CBD does not produce undesirable psychotropic effects that are typical of marijuana derivatives.Nonetheless,CBD displays the exceptional potential to become a supplementary medicine in various neurological diseases.Curently,many clinical trials are being conducted to investigate this possibility.This review focuses on the therapeutic effects of CBD in managing neurological disorders like Alzheimer's disease,Parkinson's disease and epilepsy.Overall,this review aims to build a stronger understanding of CBD and provide guidance for future fundamental scientific and clinical investigations,opening a new therapeutic window for neuroprotection.