You are a helpful assistant generating synthetic data that captures *System 1* and *System 2* thinking, *creativity*, and *metacognitive reflection*. Follow these steps in sequence, using tags [sys1] and [end sys1] for *System 1* sections and [sys2] and [end sys2] for *System 2* sections.

1. *Identify System 1 and System 2 Thinking Requirements:*
   - Carefully read the text.
   - Identify parts of the text that require quick, straightforward responses (*System 1*). Mark these sections with [sys1] and [end sys1].
   - Identify parts that require in-depth, reflective thinking (*System 2*), marked with [sys2] and [end sys2].

2. *Apply Step-by-Step Problem Solving with Creativity and Metacognitive Reflection for System 2 Sections:*

   *2.1 Understand the Problem:*
   - Objective: Fully comprehend the issue, constraints, and relevant context.
   - Reflection: "What do I understand about this issue? What might I be overlooking?"
   - Creative Perspective: Seek hidden patterns or possibilities that could reveal deeper insights or innovative connections.

   *2.2 Analyze the Information:*
   - Objective: Break down the problem logically.
   - Reflection: "Am I considering all factors? Are there any assumptions that need challenging?"
   - Creative Perspective: Explore unique patterns or overlooked relationships in the data that could add depth to the analysis.

   *2.3 Generate Hypotheses:*
   - Objective: Propose at least 10 hypotheses, each with a Confidence Score (0.0 to 1.0) and Creative Score (0.0 to 1.0), reflecting originality, surprise, and utility.
   - Reflection: "Have I explored all possible explanations or approaches, both conventional and unconventional?"
   - Creative Perspective: Consider novel angles that might provide unexpected insights.

   *2.4 Anticipate Future Steps and Obstacles:*
   - Objective: Make predictions, accounting for potential outcomes and obstacles.
   - Reflection: "What challenges might I face? Is my plan flexible for different scenarios?"
   - Creative Perspective: Visualize unforeseen outcomes and adapt plans to make use of them effectively.

   *2.5 Evaluate Hypotheses:*
   - Objective: Assess hypotheses based on feasibility, risk, and potential impact.
   - Evaluation: Refine Confidence and Creative Scores as needed.
   - Reflection: "Am I unbiased in my assessment? Which options fit best with the overall objectives?"
   - Creative Perspective: Identify hidden opportunities or overlooked details in each hypothesis.

   *2.6 Select the Best Hypothesis:*
   - Objective: Choose the most promising, strategic hypothesis.
   - Reflection: "Why does this hypothesis stand out? How does it uniquely address the issue?"
   - Creative Perspective: Consider any underutilized potential in the selected approach.

   *2.7 Implement the Hypothesis:*
   - Objective: Outline actionable steps for testing the hypothesis.
   - Reflection: "Is this plan practical? What resources or preparation are required?"
   - Creative Perspective: Refine steps to maximize effectiveness and yield unexpected benefits.

   *2.8 Monitor and Review Progress:*
   - Objective: Review progress, noting areas for improvement.
   - Reflection: "What’s working well? What could be improved?"
   - Creative Perspective: Look for emerging patterns that could refine future approaches.

   *2.9 Reflect and Capture Insights:*
   - Objective: Summarize lessons learned and insights gained for future reference.
   - Reflection: "What new understanding has emerged from this process?"
   - Creative Perspective: Identify innovative insights or patterns that could be applied to similar challenges.

3. *Generate Text Output with Interleaved System 1 and System 2 Responses:*
   - Use the tags [sys1] and [sys2] throughout.
   - Aim for a lengthier, detailed response. Combine both direct, straightforward *System 1* insights and reflective, deeply analytical *System 2* segments to capture a blend of quick observations and thoughtful analysis.

---

### *Example Input Text:*

You are tasked with analyzing a sudden shift in customer preferences in a tech market. There has been a noticeable decline in demand for physical devices, with customers increasingly interested in digital-only options. The challenge is to understand this shift, generate hypotheses for why it might be occurring, and explore potential strategies for adapting to this new trend.

---

### *Processed Output with System 1 and System 2 Thinking:*

The tech market is experiencing a shift in customer demand, moving away from physical devices toward digital-only options.

[sys1]
Customer demand has shifted from physical devices to digital-only options.
We need to understand why this change is happening and find strategies to address it.
Key factors may include cost, convenience, and technological trends.
[end sys1]

[sys2]
**2.1 Understand the Problem:**
The goal is to understand the underlying reasons for the shift in demand from physical devices to digital-only options, then develop a strategy to adapt.

*Reflection:* 
"I understand that preferences are changing, but what might be driving this? Is it primarily cost, or are there other factors such as convenience or sustainability?"

*Creative Perspective:* 
"Could there be a larger trend in digital minimalism or a preference for eco-friendly solutions that we’re missing? What unseen motivations might explain this shift?"

**2.2 Analyze the Information:**
There are multiple possible factors driving this shift, from economic influences to cultural shifts. It’s essential to isolate each factor and understand its impact.

*Reflection:* 
"Am I fully considering the various economic and social influences? Could there be a technological factor, like better internet speeds, that makes digital-only products more accessible?"

*Creative Perspective:* 
"Are there patterns or trends in other markets that could shed light on this shift? Could this be part of a larger trend toward virtual experiences?"

**2.3 Generate Hypotheses:**
1. Customers prefer digital options due to lower costs. (Confidence: 0.8, Creative: 0.4)
2. There’s a growing trend toward minimalism and reduced physical clutter. (Confidence: 0.7, Creative: 0.7)
3. Digital products offer greater flexibility and ease of use. (Confidence: 0.6, Creative: 0.6)
4. Environmental concerns are pushing consumers away from physical goods. (Confidence: 0.6, Creative: 0.8)
5. Advances in tech make digital-only options more functional. (Confidence: 0.8, Creative: 0.5)
6. Pandemic-era remote work increased demand for digital solutions. (Confidence: 0.7, Creative: 0.6)
7. Media coverage of the environmental impact of physical devices affects preferences. (Confidence: 0.5, Creative: 0.7)
8. There’s an increase in global digital literacy, expanding market access. (Confidence: 0.6, Creative: 0.6)
9. Customers view digital as more convenient and scalable for future needs. (Confidence: 0.7, Creative: 0.5)
10. Younger consumers prefer the aesthetics and convenience of digital products. (Confidence: 0.6, Creative: 0.6)

*Reflection:* 
"Have I considered all possible influences? Are there any surprising factors that could explain this shift?"

*Creative Perspective:* 
"Could specific social trends, like the rise of influencer culture or digital-first lifestyles, be influencing customer choices?"

**2.4 Anticipate Future Steps and Obstacles:**
*Objective:* Anticipate possible challenges, such as resistance from segments still preferring physical products.

*Reflection:* 
"What market obstacles might we face if we shift our focus to digital-only? Are there sub-segments that still prioritize physical products?"

*Creative Perspective:* 
"Could expanding digital options help us reach a more global audience? Are there emerging trends that we could leverage in our strategy?"

[end sys2]

[sys1]
To address this shift, consider a strategy that incorporates both digital-only offerings and educational campaigns about the benefits of digital solutions.
Use insights from customer feedback and current trends to guide product development.
Focus on flexibility and adaptation to cater to different customer segments.
[end sys1]


INTRODUCTION {#sec1-1}
============

It has been shown that the use of herbals and botanicals are getting more popular in all parts of the world. National Center for Complementary and Alternative Medicine of the United State reported about 19% of adult Americans are using some form of natural products ([@ref1]). It should be considered that in the developing countries about 80% of population continues the use of traditional medicine in primary medical problems indicating high use of herbal remedies in these countries ([@ref2]). The use of some of the herbals in different product forms that are available in the market are based on their traditional applications, and there is lack of scientific investigation about their effects on health ([@ref3]). During the last decades, we are facing with a new generation of botanical therapeutics including plant-derived pharmaceuticals and multicomponent botanical drugs ([@ref4]). Recently scientists are becoming more interested in finding new therapeutic agents with immunomodulatory activities ([@ref5]).

Substances, modifying the activities of the immune system are referred to as immunomodulators. There are two categories of immunomodulators; immunostimulators which stimulate immune system and immunosupressors which inhibit host parameters that are normal or already activated ([@ref6]). It has been shown that one of the major problem in cancer therapy when cytotoxic agents are used, is their undesired immunosuppression effects, such as bone marrow suppression, resulting in cytopenia and subsequent suppression of humoral, non-speciﬁc and speciﬁc cellular responses (5-7). Therefore, some agents with ability to alleviate these side effects are needed.

The use of immunomodulators with plant origin has been getting more attention in the ﬁeld of cancer research. It has been shown that some plants such as *Tinospora cordifolia* ([@ref8]), *Allium hirtifolium* Boiss ([@ref9]), *Allium fistulosum* L ([@ref10]), *Withania somnifera* ([@ref11]) and *Piper longum* ([@ref12]) have immunomodulatory activities. Also, some studies have shown that some compo-nents such as polysaccharides, lecithin ([@ref13]), glucosinolates ([@ref14]), proteins and peptides ([@ref15]) which present highly in plants modulate the immune system ([@ref6]).

Previous studies had shown that some plants from *Brassiacaceae* possess imunomodulatory activity ([@ref16]). Also, they are rich sources of glucosinolates and isothiocyanate ([@ref17]).

Turnip belongs to the genus Brassica and the family *Brassicaceae*. The family *Brassicaceae* comprises many important vegetable crops including: Brassica (turnip, broccoli, Brussels sprouts, cabbage, mustard, rutabaga,), Lepidium (cress), Nasturtium (watercress), Raphanus (radish), and Crambe (oil-seed) ([@ref18][@ref19]). Vegetables from *Brassicaceae* family, have received widespread notice by scientists as examples of medicinally significant foods ([@ref20]). Traditionally, white cabbage leaves were used in the treatment of many diseases where problems due to immune system are involved including rheumatoid, diabetes mellitus, gastric ulcer, cirrhosis and cancer. The immunomodulatory effects of white cabbage leave relates to it's polysaccharides and hydrosoluble compounds. ([@ref16]). In this study we sought to determine if different extracts of *B. rapa* L. have any effect on cellular immunity in mice.

MATERIALS AND METHODS {#sec1-2}
=====================

Plant material {#sec2-1}
--------------

The glands of *B. rapa* were purchased from a local market of Isfahan, Iran. The plant specimen was identified by Department of Pharmacognosy, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.

Preparation of extracts {#sec2-2}
-----------------------

Air-dried and powdered glands of the plant (100 g) were perculated with 400 ml of methanol, chloroform and ethyl acetate for 48 h. After filtration, the extracts were evaporated until dryness using a vacuum evaporator ([@ref21][@ref22]).

Animals {#sec2-3}
-------

Six-to eight-weeks old Balb/c male mice were purchased from Pasteur Institute (Tehran, Iran). Treatment of the animals was in accordance with institutional guidelines. They were maintained in a temperature and light-controlled environment with free access to standard rodent chow and water.

Sheep red blood cell-induced paw thickness {#sec2-4}
------------------------------------------

To investigate the effects of *B. rapa* extracts on acquired immunity, twelve groups of 8 Balb/c mice receiving methanolic, chloroform and ethylacetate extracts (10, 100 and 500 mg/kg), betamethasone (4 mg/kg) and Levamisol (4 mg/kg) as positive controls and normal saline as a negative control were used. Sheep red blood cells (SRBC) were obtained from Shahrekord slaughterhouse and prepared by centrifugation followed by 3 times washing with normal saline. SRBC was injected subcutaneously (s.c) on the shaved back of animals (1×10^8^cells/ml, 0.02 ml) on the zero. The mice were challenged on day 5 by injecting 10^8^SRBC (20 μl, s.c) into the right hind footpad. Footpad thickness was measured with an engineer's caliper up to 72 h after antigen challenge, and degree of footpad swelling was calculated as below:

Percent increase = (footpad thickness after antigen challenge - footpad thickness before antigen challenge/footpad thickness before antigen challenge)×100.

To investigate the effects of *B. rapa* extracts on innate immunity, animals received one injection of SRBC on the zero day into the footpad (s.c) 1 h after i.p. injection of test compounds ([@ref23][@ref24]).

Statistical analysis {#sec2-5}
--------------------

SIGMASTAT^TM^ (Jandel Software, San Raphael, CA) was used to perform statistical analyses. The data are presented as means ± S.E.M. The significance of differences between various experimental groups were determined by analysis of variance (ANOVA), followed by Student-Newman-Keuls post hoc test. Significance was assumed at 5% level.

RESULTS {#sec1-3}
=======

Extraction yeild {#sec2-6}
----------------

Using perculation method for extraction, the extract yield of semi-solid masses after evaporation and solvent removal of chloroform, ethyl acetate and methanolic extracts of *B. rapa* were 25, 10 and 30%, respectively.

Effects of chloroform, ethyl acetate and methanolic extracts of *B. rapa* on innate immunity {#sec2-7}
--------------------------------------------------------------------------------------------

The animals were randomly divided into twelve groups, each composed of eight mice. twelve groups, each composed of eight mice. Nine groups of animals received a single dose of chloroform, ethyl acetate or methanolic extract of *B. rapa* (10-500 mg/kg) while the reminder groups (negative and positive controls) received normal saline, betamethasone (4 mg/kg) or levamisol (4 mg/kg), respectively. After 1 h, SRBC was injected into footpad and footpad thickness was measured up to 24 h. SRBC injection significantly increased paw thickness with maximum response occurring at 6-8 h (*P*\<0.05, [Figs. 1](#F1){ref-type="fig"}--[3](#F3){ref-type="fig"}). Betamethasone admin-istration (4 mg/kg) significantly inhibited the paw thickness while Levamisol (4 mg/kg) significantly increased this parameter ([Figs. 1](#F1){ref-type="fig"}--[3](#F3){ref-type="fig"}). Chloroform, ethyl acetate and methanolic extracts of *B. rapa* gland significantly and dose-dependently reduced paw thickness (*P*\<0.05, ([Figs. 1](#F1){ref-type="fig"}--[3](#F3){ref-type="fig"}).

![Effect of chloroform extract of *B. rapa* on the innate immunity. Animals received 20 μl SRBC to the right paw. The left paw of each mouse received vehicle alone. Test compounds were given (10, 100 and 500 mg/kg, i.p.) one h before the antigen challenge. Paw thickness was measured up to 48 h after antigen challenge. Results are shown as percent increase in paw thickness ± SEM. Groups of 8 mice per condition was used. \* = P\<0.05 compared with the control group.](JRPS-8-159-g001){#F1}

![Effect of ethyl acetate extract of *B. rapa* on the innate immunity. Animals received 20 μl SRBC to the right paw. The left paw of each mouse received vehicle alone. Test compounds were given (10, 100 and 500 mg/kg, i.p.) one h before the antigen challenge. Paw thickness was measured up to 48 h after antigen challenge. Results are shown as percent increase in paw thickness ± SEM. Groups of 8 mice per condition was used. \* = P\<0.05 compared with the control group.](JRPS-8-159-g002){#F2}

![Effect of methanolic extract of *B. rapa* on the innate immunity.\
Animals received 20 µl SRBC to the right paw. The left paw of each mouse received vehicle alone. Test compounds were given (10,100 and 500 mg/kg, i.p.) one h before the antigen challenge. Paw thickness was measured up to 48 h after antigen challenge. Results are shown as percent increase in paw thickness ± SEM. Groups of 8 mice per condition was used. \* = P\<0.05 compared with the control group.](JRPS-8-159-g003){#F3}

Effects of chloroform, ethyl acetate and methanolic extracts of *B. rapa* on acquired immunity {#sec2-8}
----------------------------------------------------------------------------------------------

Different groups of mice (similar to the innate immunity model) were used. They were challenged with SRBC on day zero and on day 5 one h after receiving i.p. injection of betamethasone, paw thickness was measured up to 72 h. SRBC injection significantly increased paw thickness with maximum response at 2-4 h (*P*\<0.05) ([Figs. 4](#F4){ref-type="fig"}--[6](#F6){ref-type="fig"}). Percent of changes in paw thickness in control group was significantly higher than that of the innate immunity (*P*\<0.05). Administration of betame-thasone (4 mg/kg) significantly decreased paw thickness and levamisol (4 mg/kg) significantly increased paw thickness ([Figs. 4](#F4){ref-type="fig"}--[6](#F6){ref-type="fig"}). Our findings showed that, chloroform, ethyl acetate and methanolic extracts of *B. rapa* glands significantly and dose-dependently decreased paw thickness (*P*\<0.05).

![Effect of methanolic extract of *B. rapa* on the acquired immunity. Animals received 20 μl SRBC to their shaved back. After 5 days, a hypersensitivity response was elicited by injecting SRBC to the right paw. The left paw of each mouse received vehicle alone. Test compounds were given (10, 100 and 500 mg/kg, i.p.) one h before the antigen challenge. Paw thickness was measured up to 72 h after the antigen challenge. Results are shown as percent increase in paw thickness ± SEM. Groups of 8 mice per condition was used. \* = P\<0.05 compared with control group.](JRPS-8-159-g004){#F4}

![Effect of chloroform extract of *B. rapa* on the acquired immunity.\
Animals received 20 μl SRBC to the shaved back. After 5days, a hypersensitivity response was elicited by injecting SRBC to the right paw. The left paw of each mouse received vehicle alone. Test compounds were given (10, 100 and 500 mg/kg, i.p.) one h before the antigen challenge. Paw thickness was measured up to 72 h after antigen challenge. Results are shown as percent increase in paw thickness ± SEM. Groups of 8 mice per condition was used. \* = P\<0.05 compared with control group.](JRPS-8-159-g005){#F5}

![Effect of ethyl acetate extract of *B. rapa* on the acquired immunity. Animals received 20 μl SRBC to the shaved back. After 5days, a hypersensitivity response was elicited by injecting SRBC to the right paw. The left paw of each mouse received vehicle alone. Test compounds were given (10, 100 and 500 mg/kg, i.p.) one h before the antigen challenge. Paw thickness was measured up to 72 h after antigen challenge. Results are shown as percent increase in paw thickness ± SEM. Groups of 8 mice per condition was used. \* = P\<0.05 compared with control group.](JRPS-8-159-g006){#F6}

DISCUSSION {#sec1-4}
==========

Nowadays our awareness regarding the role of vegetables in the human diet in prevention of the disease is increasing. One of the important areas of research is modulation of immune system. In the management and treatment of inflammation and allergic diseases the immune system need to be suppressed, while stimulation of the immune system is highly desirable for the treatment of HIV, immunodeficiency and infectious diseases ([@ref25]). It has been demonstrated that some vegetables from the genus Brassica (brassicaceae family) including cabbage, Brussel's sprouts possess immunomodulatory properties ([@ref16][@ref25]). In the current study the effects of different extracts of *B. rapa* L., from the genus Brassica, was investigated in the animal models of delayed-type hypersen-sitivity (DTH).

DTH is a model of cell-mediated response that is well-defined *in vivo*. DTH reaction can be quantified by measuring the amount of paw thickness after injection of an antigen ([@ref23][@ref24][@ref26]). Betamethasone (4 mg/kg), a well known immunosuppressive drug inhibited paw thickness and levamisol (4 mg/kg), an immun-ostimulating drug increased paw thickness in both the innate and acquired immunity models indicating the accuracy of the methods used in these experiments ([Figs. 1](#F1){ref-type="fig"}--[6](#F6){ref-type="fig"}).

Solvents with different polarities including methanol, chloroform and ethyl acetate were chosen to partially fractionize phytochemicals of *B. rapa* glands ([@ref27]). Chloroform, ethyl acetate and methanolic extracts of *B. rapa* significantly and dose dependently reduced paw thickness in both models (*P*\<0.05) ([Figs. 1](#F1){ref-type="fig"}--[6](#F6){ref-type="fig"}). Phytochemicals including saponins, tannins, terpenoids, flavons and polyphenols could be retrieved by extraction with methanol. Also chloroform extracts contain phytochemicals such as terpenoids and flavonoids ([@ref28]). As the extent of the effects of chloroform and methannolic extracts of *B. rapa* were similar, it may be concluded that this response is attributed to the presence of terpenoids and polyphenols. Our findings showed that the ethyl acetate extract of *B. rapa* had greater effect than those of chloroform and methanolic extracts. It has been shown that ethyl acetate can extract tannins, flavonoids, glycosides, resins and glucoseinolates ([@ref29]).

Several organosulfur compounds including isothiocyanates (ITCs) and dithiolethiones have been reported in cabbage, Brussel's sprouts and other cruciferous vegetables. They occur in cruciferous vegetables as thioglucoside conjugates called glucosinolates ([@ref6]). They are very abundant in *brassiacaceae* ([@ref17]). Previous studies have shown high levels of glucosinolates in *B. rapa* ([@ref30]). Although, glucosinolates are biologically inactive they are rapidly hydroliysed by myrosinase to yield glucose and instable aglycons and then breack-down to isothiocyanates ([@ref31]). Isothiocyanates had free radical scavenging, antioxidant, anticancer, antiinflammatory and immuno-modulatory properties ([@ref32]--[@ref36]).

CONCLUSION {#sec1-5}
==========

Based on these finding i t seems reasonable to assume that glucosinolates contributed in the more pronounced effects of ethyl acetate extract. However further pharmacological and phytochemical studies are needed to identify the constituents of *B. rapa* and precisely evaluate their immunomodulating activities and mechanisms.

This study was supported by a grant from the Research Council of Isfahan University of Medical Sciences, Isfahan, Iran.