Production of autotetraploid plants by in vitro chromosome engineering in Allium hirtifolium

Persian shallot has been propagated vegetatively through daughter bulbs and has a narrow genetic base.In the present study,to create genetic diversity in terms of ploidy level,the effects of in vitro polyploidization were investigated on morphological,physiological and phytochemical traits.Different colchicine treatments(0,0.3%,0.5% and 0.7% concentrations)for 24,36 and 48 h and oryzalin treatments(0,0.001%,0.002% and 0.004% concentrations)for 4,6 and 8 h were used.A sterile basal plate of bulbs was used for the induction of polyploidy in liquid and solid media.After obtaining plantlets,root tip chromosomes were counted for the determination of ploidy levels.Flow cytometry was performed to confirm the chromosome counting results.The regenerated diploid and tetraploid plantlets were transferred to ex vitro conditions.Colchicine and oryzalin were both successful in inducing polyploids and the polyploids induced were tetraploids and mixoploids.The highest induction of polyploidy was obtained in solid media using 0.5%(w/v)colchicine for 36 h(35.0% polyploidy induction)and in 0.001%(w/v)Oryzalin for 8 h(45.5% polyploidy induction).Differences in plantlet height and weight,bulblet weight,density,stomatal width and length,and chlorophyll content were observed between tetraploid and diploid plants.Chromosome duplication,as a result of tetraploid induction,significantly influenced the antioxidant content and enzyme activities.The results showed that total phenolic content,allicin,and antioxidant capacity were significantly higher in the tetraploid plantlets than that in the diploid plantlets.SRAP loci polymorphisms indicated that the 12 autotetraploid plants(with high allicin content)had different genotypes from the parental diploid plant.Besides,in vitro polyploidy induction not only duplicated chromosomes but also altered the DNA sequence in Allium hirtifolium.In vitro induction of tetraploids in A.hirtifolium can be a reliable way to obtain suitable plant material for breeding programs to generate new genotypic variations.

Evaluation of the effect of shodhana(detoxification)process using chromatographic profiling(HPTLC,HPLC,LC-MS,and GC-MS)and estimation of the content of toxic colchicine in Gloriosa superba Linn.tubers

Shodhana(purification/detoxification)is an important aspect of Ayurveda.It has been described in several important pieces of literature.In this study,we performed shodhana process on Langali(Gloriosa superba Linn.)tubers of family Liliaceae,a semi-woody herbaceous climber with significant therapeutic properties.We compared the unprocessed(before purification)and processed(after purification)samples based on the phytochemical,physicochemical,and chromatographic study,including HPTLC,HPLC,GC-MS and LC-MS.Significant changes were observed in profiling after the shodhana process.This plant contains colchicine,known for its toxic effects,particularly cardiotoxicity.Therefore,we quantified colchicine in the unprocessed and processed samples of Gloriosa superba Linn.tubers using HPLC method.Results showed that the percentage of colchicine was reduced by 13.67%in chloroform extract and 20.97%in ethanol extracts after the shodhana process.

Doubling Efficiency of Maize Haploids Treated with Different Methods

Haploid seedlings were inducted from different maize materials.At 2-3-leaf stage,maize haploids were treated with 0.06% colchicine and 2.0% dimethyl sulfoxide（DMSO） by dipping root,dripping heart leaf and acupuncturing growing point,respectively.The doubling rate and mortality rate in different treatments were analyzed by variance analysis and multiple comparisons.The result showed that growing point acupuncturing method exhibited the highest doubling efficiency with an average doubling rate of 23%,seed-setting rate of 21.4%,and mortality rate of 16.3%.Composed with other two chemical doubling methods,growing point acupuncturing method significantly improved the doubling rate of maize haploids with a lower application dose of colchicine.This study laid the foundation for industrial application of haploid breeding techniques.

Colchicine-induced Melilotus Research

[Objective] The aim was to identify the ploidy of sweet clover induced by colchicine. [Method] The sweet clover germinating seeds and colchicine solution were used for materials to research colchicine-induced Melilotus. [Result] The results showed that the concentration of 0.2% colchicine solution could induced double sweet clover which showed corresponding features of polyploidy plants as a whole showed great features. [Conclusion] Sweet clover induced by colchicine was polyploidy.

Colchicine Inhibited the Expression of Tissue Inhibitor of Metalloprotenase-1 and Interleukin-6 in Cultured Activated Hepatic Stellate Cells

Cultured HSCs were treated colchicine with different concentrations for 12 h, respectively. The effects of eolehicine on HSCs growth were measured by MTT assay. Effects of eolchicine on gene expression of HSCs were analysed by using a self-made oligonucleotide mieroarray. Colehieine inhibited HSCs growth in a dose-dependent manner. After 12 h of treatment with 6.25 mg/L of colehicine, the expression of tissue inhibitor of metalloprotenase-1 （TIMP-1） and the expression of interleukin-6 （IL-6） in HSCs were downregulated by 2.3 folds and 2.1 folds, respectively. These results suggest that colchieine＇s beneficial effects may, at least in part, owe to the inhibitory to the proliferation of HSCs and down regulation of the expression of both TIMP1 and IL-6 in HSCs.

Study on the Polyploid Induction of Lycium ruthenicum Murr.

With apical leaves of Lycium ruthenicum Murr as experimental material, the polyploids of L. ruthenicum were induced with colchicine solution, and total four polyploidy plants were identified by chromosome courts. The results showed that tetraploid plants of L. ruthenicum were successfully induced with 300 mg/L of colchicine solution after 7 and 21 d of induction, with 400 mg/L of colchicine solution after 14 days of induction, and with 500 mg/L of colchicine solution after 7 days of induction, respectively.

Colchicine reduces procollagen Ⅲ and increases pseudocholinesterase in chronic liver disease

AIM:To test whether colchicine would be an effective antif ibrotic agent for treatment of chronic liver diseases in patients who could not be treated with α-interferon.METHODS:Seventy-four patients(46 males,28 females) aged 40-66 years(mean 53±13 years) participated in the study.The patients were affected by chronic liver diseases with cirrhosis which was proven histologically(n=58);by chronic active hepatitis C(n=4),chronic active hepatitis B(n=2),and chronic persistent hepatitis C(n=6).In the four patients lacking histology,cirrhosis was diagnosed from anamnesis,serum laboratory tests,esophageal varices and ascites.Patients were assigned to colchicine(1 mg/d) or standard treatment as control in a randomized,double-blind trial,and followed for 4.4 years with clinical and laboratory evaluation.RESULTS:Survival at the end of the study was 94.6% in the colchicine group and 78.4% in the control group(P=0.001).Serum N-terminal peptide of type Ⅲ procollagen levels fell from 34.0 to 18.3 ng/mL(P=0.0001),and pseudocholinesterase levels rose from 4.900 to 5.610 mU/mL(P=0.0001) in the colchicine group,while no signif icant change was seen in controls.Best results were obtained in patients with chronic hepatitis C and in alcoholic cirrhotics.CONCLUSION:Colchicine is an effective and safe antifibrotic drug for long-term treatment of chronic liver disease in which fi brosis progresses towards cirrhosis.

Pulmonary delivery of liposomal dry powder inhaler formulation for effective treatment of idiopathic pulmonary fibrosis

Dry powder inhaler Liposomes were prepared to investigate the effectiveness of pulmonary delivery of Colchicine and Budesonide for Idiopathic Pulmonary fibrosis. Budesonide(BUD) and Colchicine(COL) liposomes were prepared by thin layer film hydration method(TFH) using 1,2-Dipalmitoyl-sn-glycero-3-phosphoglycerol sodium(DPPG), Hydrogenated Soyaphosphotidylcholine(HSPC), Soyaphosphatidylcholine(SPC), cholesterol(CHOL) and drug in different weight ratios. The optimum lipid composition for BUD(74.22 ± 0.97%) was DPPG:HSPC: CHOL(4:5:1) and for COL(50.94 ± 2.04%) was DPPG: SPC: CHOL(3:6:1). These compositions retained drug for a longer period of time so selected for further study. Liposomes were found to be spherical in shape with mean size below 100 nm. Liposomes lyophilized using Mannitol as carrier and cryoprotectant showed high entrapment efficiency(97.89-98.6%). The powder was dispersed through an Andersen cascade impactor to evaluate the performance of the aerosolized powder. It was found that prepared liposomal dry powder inhaler(DPIs) sustained the drug release up to 24 hours. Optimized Budesonide DPI Formulation B2(86.53 ± 1.9%), Colchicine DPI Formulation C2(90.54 ± 2.3 %) and BUD and COL DPI Combination M2(89.91 ± 1.8%, 91.23 ± 1.9%). Histopathological results, measurements of lung hydroxyproline content, Myeloperoxidase activity indicated that liposomal drypowder inhaler administration attenuates lung fibrosis induced by bleomycin. Long term stability studies indicated that lyophilised BUD and COL liposomes were stable for 6 months at(25 °C± 2 °C, 60% ± 5% RH) and refrigerated conditions(2-8 °C). These results supported that combination of budesonide and colchicine liposomal dry powder inhaler pulmonary drug delivery for treatment of idiopathic Pulmonary Fibrosis exhibits prolonged drug retention at targeted site and reduces the systemic exposure.

Mechanical properties of hepatocellular carcinoma cells

AIM: To study the viscoelastic properties of human hepatocytes and hepatocellular carcinoma (HCC) cells under cytoskeletal perturbation, and to further to study the viscoelastic properties and the adhesive properties of mouse hepatoma cells (HTC) in different cell cycle. METHODS: Micropipette aspiration technique was adopted to measure viscoelastic coefficients and adhesion force to collagen coated surface of the cells. Three kinds of cytoskeleton perturbing agents, colchicines (Col), cytochalasin D (CD) and vinblastine (VBL), were used to treat HCC cells and hepatocytes and the effects of these treatment on cell viscoelastic coefficients were investigated. The experimental results were analyzed with a three-element standard linear solid. Further, the viscoelastic properties of HTC cells and the adhesion force of different cycle HTC cells were also investigated. The synchronous G(1) and S phase cells were achieved through thymine-2-desoryriboside and colchicines sequential blockage method and thymine-2-desoryriboside blockage method respectively. RESULTS: The elastic coefficients, but not viscous coefficient of HCC cells (K(1)=103.6+/-12.6N.m(-2), K(2)=42.5 +/ 10.4N.m(-2), mu=4.5 +/- 1.9Pa.s), were significantly higher than the corresponding value for hepatocytes (K(1)=87.5 +/- 12.1N.m(-2), K(2)=33.3+/-10.3N.m(-2), mu=5.9+/-3.0Pa.s, P【0.01). Upon treatment with CD, the viscoelastic coefficients of both hepatocytes and HCC cells decreased consistently, with magnitudes for the decrease in elastic coefficients of HCC cells (K(1): 68.7 N.m(-2) to 81.7N.m(-2), 66.3% to 78.9%; K(2): 34.5N.m(-2) to 37.1N.m(-2), 81.2% to 87.3%, P【0.001) larger than those for normal hepatocytes (K(1): 42.6N.m(-2) to 49.8N.m(-2), 48.7% to 56.9%; K(2): 17.2N.m(-2) to 20.4N.m(-2), 51.7% to 61.3%, P【0.001). There was a little decrease in the viscous coefficient of HCC cells (2.0 to 3.4Pa.s, 44.4 to 75.6%, P【0.001) than that for hepatocytes (3.0 to 3.9Pa.s, 50.8 to 66.1% P【0.001). Upon treatment with Col and VBL, the elastic coefficients of hepatocytes generally increased or tended to increase while those of HCC cells decreased. HTC cells with 72.1% of G(1) phase and 98.9% of S phase were achieved and high K(1), K(2) value and low mu value were the general characteristics of HTC cells. G(1) phase cells had higher K(1) value and lower mu value than S phase cells had, and G(1) phase HTC cells had stronger adhesive forces ((275.9 +/- 232.8) x 10(-10)N) than S phase cells ((161.2 +/- 120.4) x 10(-10)N, P【0.001). CONCLUSION: The difference in both the pattern and the magnitude of the effect of cytoskeletal perturbing agent on the viscoelastic properties between HCC cells and hepatocytes may reflect differences in the state of the cytoskeleton structure and function and in the sensitivity to perturbing agent treatment between these two types of cells. Change in the viscoelastic properties of cancer cells may affect significantly tumor cell invasion and metastasis as well as interactions between tumor cells and their micro-mechanical environments.

Tolerance of nestin^+ cholinergic neurons in the basal forebrain against colchicine-induced cytotoxicity

In the present study we injected colchicine into the lateral ventricle of Sprague-Dawley rats to investigate the effects of colchicine on the number of different-type neurons in the basal forebrain and to search for neurons resistant to injury. After colchicine injection, the number of nestin^＋ cholinergic neurons was decreased at 1 day, but increased at 3 days and peaked at 14-28 days. The quantity of nestincholinergic neurons, parvalbumin-positive neurons and choline acetyl transferase-positive neurons decreased gradually. Our results indicate that nestin^＋ cholinergic neurons possess better tolerance to colchicine-induced neurotoxicity.

Extraction and Anti-inflammatory Effects of Colchicine from Sagittaria sagittifolia

[Objective] This study was to study the optimal extraction technology and anti-inflammatory effects of colchicine from Sagittaria sagittifolia. [Method] The ef- fects of ethanol concentration, extraction time, extraction temperature and solid-liquid ratio on the extraction rate of colchicine from S. sagittifolia were investigated. On the basis of single-factor experiments, an L9 （34） orthogonal test was carried out to optimize the extraction process. According to the optimal extraction process, the content of colchicine in S. sagittifolia was determined by high performance liquid chromatography. The anti-inflammatory ability of colchicine was evaluated through an anti-inflammatory test in vitro. [Result] The optimal extraction process of colchicine from S. sagittifolia was as follows： ethanol concentration of 60%, extraction temper- ature of 50℃, extraction time of 30 min, and solid-liquid ratio of 1：25 （g/ml）. The content of colchicine in S. sagittifolia was determined as 40.58 μg/100 mg. Com- pared with the control, the colchicine from S. sagittifolia （9.0 and 4.5 g/kg） inhibited the increase in PGE2, TNF-α and IL-1β contents in pleural fluid （P〈0.05）. High-dose colchicine inhibited the increase in TNF-α, IL-1β and MDA contents in lung tissue （P〈0.01）, and middle-dose colchicine inhibited the increase in IL-1β content in lung tissue （P〈0.01）. [Conclusion] The colchicine in S. sagittifolia has a good anti-inflam- matory effect, which may be achieved through hindering the production of inflam- matory mediators and antioxidation.

Development of a novel rat model of heterogeneous hepatic injury by injection with colchicine via the splenic vein

AIM To develop a novel rat model of heterogeneous hepatic injury.METHODS Seventy male Sprague-Dawley rats were randomly divided into a control group(n = 10) and a colchicine group(n =60). A 0.25% colchicine solution(0.4 mL/kg) was injected via the splenic vein in the colchicine group to develop a rat model of heterogeneous hepatic injury. An equal volume of normal saline was injected via the splenic vein in the control group. At days 3, 7, and 14 and weeks 4, 8, and 12 after the operation, at least seven rats of the colchicine group were selected randomly for magnetic resonance imaging(MRI) examinations, and then they were euthanized. Ten rats of the control group underwent MRI examinations at the same time points, and then were euthanized at week 12. T2-weighted images(T2 WI) and diffusion weighted imaging(DWI) were used to evaluate the heterogeneous hepatic injury. The heterogeneous injury between the left and right hepatic lobes was assessed on liver sections according to the histological scoring criteria, and correlated with the results of MRI study. RESULTS Obvious pathological changes occurred in the hepatic parenchyma in the colchicine group. Hepatic injury scores were significantly different between the left and right lobes at each time point(P < 0.05). There was a significant difference in apparent diffusion coefficient(ADC) of DWI and liver-to-muscle ratio(LMR) of T2 WI between the left and right lobes of rats in the colchicine group(P < 0.05) at each time point, and similar results were observed between the colchicine and control groups. Besides, there was a significant correlation between hepatic injury scores and ADC values or LMR(r =-0.682, P = 0.000; r =-0.245, P = 0.018).CONCLUSION Injection with colchicine via the splenic vein can be used to successfully develop a rat model of heterogeneous hepatic injury. DWI and T2 WI may help evaluate the heterogeneous injury among liver lobes.

<i>In Vitro</i>Induction of Polyploidy in <i>Citrus reticulata</i>Blanco

Possibility of polyploidy induction by colchicine in “Balady” mandarins was investigated in vitro. Seeds were immersed in different concentrations of colchicine solutions (0.01%, 0.05%, 0.1% and 0.2%) for different durations (12, 24 and 48 hr), then cultured in vitro on MS medium at half strength. Seed survival percentage decreased by increasing colchicine concentration and duration of treatment showing the lowest percentage at 0.2% for 48 hr. The highest DNA content was recorded at 0.2% for 24 hr. Stomata No. per unit area was decreased by colchicine treatments;moreover stomata length and width were studied. The results indicated that colchicine treatment at 0.1% for 48 hr had the highest tetraploid induction efficiency percentage.

Polyploidy Induction of Pteroceltis tatarinowii Maxim.

[Objective] This study was conducted to obtain tetraploid Pteroceltis tatar# nowii Maxim. with excellent ornamental traits. [Method] The stem apex growing points of Pteroceltis tatarinowii Maxim. were treated with different concentrations of colchicine solution for different hours to figure out a proper method and obtain poly- ploids. [Result] The most effective induction was obtained by treatment with 0.6%- 0.8% colchicine for 72 h with 34.2% mutation rate. Flow cytometry and chromosome observation of the stem apex growing point of P. tatarinowii Maxim. proved that the tetraploid plants were successfully obtained with chromosome number 2n=4x=36. [Conclusion] The result not only fills the blank of polyploid breeding of P. tatarinowii, but also provides an effective way to broaden the methods of cultivation of fast- growing, high-quality, disease-resilience, new varieties of Pteroceltis.

Genome-wide analysis of microRNAs induced and regulated by colchicine in mulberry

A variety of plants were colchicine treated to double their chromosome number.Chromosomes are genetic carriers that determine the individual traits of organisms.The doubling of chromosomes will lead to modifications in plant morphology,physiology and genetics.To determine the response of mulberry trees induced by colchicine,using mulberry variety Yu-711 leaves as research materials,two small RNA libraries(control and experimental groups)were constructed.It was found that 45 known miRNA genes and 78 predicted novel miRNA genes in the sequence results.A comparison of data between the control group and the experimental group revealed 37 differentially expressed miRNA genes,19 genes of which were up-regulated and 18 genes of which were down-regulated.Eight miRNAs were selected from 37 differentially expressed miRNA genes.These miRNAs were verified by quantitative real-time PCR,and the expression levels of these miRNAs were found to be consistent with those obtained by sequencing,which proved the accuracy of sequencing results.The PsRNATarget software was used to predict the target genes of 8 miRNAs.The Gene Ontology and Kyoto Encycopedia of Genes and Genomes analysis were used to collect the functions of target genes and confirm that target genes are mainly involved in biological processes,cell components and molecular functions.novel-77 miRNA was selected from 37 differentially expressed miRNAs.What will serve as a fundamental data in understanding mulberry miRNAs function in molecular biology research and further provides the molecular mechanism of mulberry gene regulation as induced by colchicine is that the prediction of novel_77 precursor sequence and the target gene(XM_010104258.2),as well as the secondary structure analysis.

Colchicine sensitizes human hepatocellular carcinoma cells to damages caused by radiation

AIM： We studied the effect of colchicine combined with radiation on the survival of human hepatocellular carcinoma （HCC） HA22T/VGH cells.METHODS： Twenty-four hours after treatment with 0-8 ng/mL colchicine, HA22T/VGH cells were irradiated at various doses （0, 1, 2, 4, and 8 Gy）. Colony assay was performed to assess the surviving cell fraction. Survival curves were fitted by using a linear-quadratic model to estimate the sensitizer enhancement ratio （SER）. Flow cytometry was used for cell cycle analysis.RESULTS： Colchicine at lower concentrations （1 and 2 ng/mL） had obvious synergy with radiation to inhibit HCC cell growth, whereas higher concentrations （4 and 8 ng/mL） had only additive effect to radiation. Pretreatment with 1 and 2 ng/mL colchicine for 24-h enhanced cell killing by radiation with SERs of 1.21 and 1.53, respectively.G2/N arrest was only observed with higher colchicine doses （8 and 16 ng/mL） after 24-h treatment, this effect was neither seen with lower doses （1, 2, and 4 ng/mL）nor with any dose after only 1 h of treatment.CONCLUSION： Our results suggest that colchicine has potential as an adjunct to radiotherapy for HCC treatment.Lower doses of colchicine possess radiosensitizing effects via some mechanism other than G2/M arrest. Further study is necessary to elucidate the mechanism.

Acute and recurring pericarditis: More colchicine,less corticosteroids

Acute and recurring pericarditis are frequently encountered clinical entities.Given that severe complications such as tamponade and constrictive pericarditis occur rarely,the majority of patients suffering from acute pericarditis will have a benign clinical course.However,pericarditis recurrence,with its painful symptoms,is frequent.In effect,recent studies have demonstrated a beneficial role of colchicine in preventing recurrence,while also suggesting an increase in recurrences with the use of corticosteroids,the traditional first-line agent.

Induced Polyploidy via Colchicine Treatment Increases Flower Size and Fruit Weight in Cape Gooseberry(Physalis peruviana L.)

An induced polyploid plant through colchicine treatment offers probably the best scope for improvement in flower size and fruit weight.Thus,in the present study,an attempt was made to induce polyploidy in Cape gooseberry using colchicine with the objective of creating more genetic variability.The colchicine concentrations were used as 0.10%(C1),0.20%(C2)and 0.40%(C3)for the duration 12(H1),24(H2)and 36(H3)hours for each concentration with seedling apex dip method(M1),cotton plug method(M2)and lanolin paste method(M3).The plants treated with 0.10%of colchicine by cotton plug method for 12 h showed the better performance during the years 2017-2018 and 2018-2019 in respect of more delay in the flower bud emergence(54 d and 53 d from the date of transplanting),anthesis(19 d and 20 d from the first appearance of bud to full anthesis of flower)and fruit setting(8.00 d and 9.00 d from the date of anthesis),bigger flower size(2.93 cm2 and 3.00 cm2)than the untreated plants.The lower percentage of pollen viability(40.33%and 40.67%)was noticed in the same treatment in comparison to control(70.33%and 72.33%).The fruit maturity was also extended(59 d and 60 d from the date of fruit set)with bigger sized fruits(length:2.53 cm and 2.57 cm,breadth:2.27 cm and 2.33 cm)as well as more fruit weight(8.70 g and 8.33 g)by the application of colchicine at 0.10%with cotton plug method for 12 h.On the basis of results obtained,the application of colchicine at 0.10%for 12 h duration with cotton plug method was found to be the best and effective treatment for induction of polyploidy as well as more flower size and fruit weight in Cape gooseberry.

A Novel Approach for Production of Colchicine as a Plant Secondary Metabolite by in Vitro Plant Cell and Tissue Cultures

The secondary metabolites synthesized by plants are economically important chemical compounds in the agricultural and industrial areas such as food, perfumery and pharmaceutical sectors. In recent years, attempts for their production by in vitro plant cell and tissue cultures have been accelerated considerably. Colchicine, the principle secondary metabolite of Colchicum autumnale L. and Gloriosa superba L., is an important alkaloid that has poison effect used for treatment of various diseases and plant breeding studies. Presently, colchicine has been produced by using the seeds of C. autumnale L. and the tubers of G. superba L. through different chemical extraction methods. Applying in vitro plant cell and tissue cultures together with metabolic and genetic engineering techniques, large-scale production of colchicine can be achieved from the above two plant species.

Psoriasis treatment: Unconventional and non-standard modalities in the era of biologics

Psoriasis is a potentially debilitating inflammatory dermatosis affecting 0.2%-4.8% of the population worldwide causing a significant occupational, personal or psychosocial morbidity to these patients for life. The basic aim of psoriasis therapy is to control the disease to maximum possible extent and improve the patient's quality of life. Management of triggers for flareups, lifestyle modifications, and dietary supplements are often recommended. Intermittent or rotational therapy with frequent alterations in treatment options is usually needed to reduce toxicity of anti-psoriatic drugs in the absence of safer alternatives. Currently, several biological agents categorized as either T-cell targeted(e.g., Alefacept, Efalizumab) or cytokine modulating(e.g., Adalimumab, Infliximab, Etanercept) are available for treating severe psoriasis. However, their high cost is often precluding for most patients. The usefulness of systemic(methotrexate, cyclosporine, acitretin or several other therapeutic agents) or topical(tar, anthralin, corticosteroids or calcipotriol ointments, phototherapy with or without psoralens) therapies has been well established for the management of psoriasis. The literature is also replete with benefits of less used non-standard and unconventional treatment modalities(hydroxycarbamide, azathioprine, leflunomide, mycophenolate mofetil, isotretinoin, fumarates, topical calcineurin inhibitors, peroxisome proliferator-activated receptors agonists, statins, sulfasalazine, pentoxifylline, colchicine, grenz ray therapy, excimer laser, climatotherapy and balneophototherapy, peritoneal dialysis, tonsillectomy, ichthyotherapy, etc.). These can be used alternatively to treat psoriasis patients who have mild/minimal lesions, are intolerant to conventional drugs, have developed side effects or achieved recommended cumulative dose, where comorbidities pose unusual therapeutic challenges, or may be as intermittent, rotational or combination treatment alternatives.