Full Communication

Immunomodulatory Effects of a Standardized Lycium barbarum Fruit Juicein Chinese Older Healthy Human Subjects

Harunobu Amagase,1 Bixuang Sun,2 and Dwight M. Nance3

1FreeLife International LLC, Phoenix, Arizona; 2Department of Nutrition, University of California at Davis, Davis, California;and 3Susan Samueli Center for Integrative Medicine, University of California at Irvine, Orange, California, USA

ABSTRACT Lycium barbarum has been traditionally used in combination with several herbs for medicinal properties, but

systematic modern clinical evaluation as a single herb has not been reported. To examine the systematic effects of L. barbarum

on immune function, general well-being, and safety, we tested the effects of a standardized L. barbarum fruit juice (GoChi�,

FreeLife International, Phoenix, AZ, USA) at 120 mL=day, equivalent to at least 150 g of fresh fruit, the amount traditionally

used, or placebo for 30 days in a randomized, double-blind, placebo-controlled clinical study in 60 older healthy adults (55–72

years old). The GoChi group showed a statistically significant increase in the number of lymphocytes and levels of interleukin-

2 and immunoglobulin G compared to pre-intervention and the placebo group, whereas the number of CD4, CD8, and natural

killer cells or levels of interleukin-4 and immunoglobulin A were not significantly altered. The placebo group showed no

significant changes in any immune measures. Whereas the GoChi group showed a significant increase in general feelings of

well-being, such as fatigue and sleep, and showed a tendency for increased short-term memory and focus between pre- and

post-intervention, the placebo group showed no significant positive changes in these measures. No adverse reactions, ab-

normal symptoms, or changes in body weight, blood pressure, pulse, visual acuity, urine, stool, or blood biochemistry were

seen in either group. In conclusion, daily consumption of GoChi significantly increased several immunological responses and

subjective feelings of general well-being without any adverse reactions.

KEY WORDS: � general well-being � humans � immunomodulation � Lycium barbarum � polysaccharides � safety

INTRODUCTION

Lycium barbarum is a Solanaceous defoliated shrub-bery, and the fruit is a famous traditional medicine in

Asian countries where it has been used for medicinal pur-poses and as a functional food for over 2,500 years. Theancient herbalist classics recorded that L. barbarum has theefficacy to nourish the liver and kidney and brighten the eye.In support of such traditional properties, modern studiesindicate that extracts from L. barbarum possess a range ofbiological activities,1–5 including immunomodulation,6–13

anti-aging, neuroprotection, antifatigue=endurance, increasedmetabolism, control of glucose and other symptoms in dia-betics, glaucoma, antioxidant efficacies, antitumor activity,and cytoprotection.

Various chemical constituents are found in L. barbarumfruit. Its reddish-orange color is derived from a group ofcarotenoids, which make up only 0.03–0.5% of the driedfruit.14 The predominant carotenoid is zeaxanthin, mainly asdipalmitate (also called physalien or physalin), making up

about one-third to one-half of the total carotenoids. Alsofound are various small molecules, such as betaine, cere-broside, b-sitosterol, and p-coumaric acid, and various vi-tamins, amino acids, taurine, and g-aminobutanoic acid,among others. K, Ca, Zn, Fe, Co, Mn, Se, Mg, and otherminerals are also present in inorganic forms.15,16

Among the chemical constituents of L. barbarum fruit, themost valuable and well-researched components are a groupof unique, water-soluble glycoconjugates—collectivelytermed L. barbarum polysaccharides (LBPs), which areestimated to make up 5–8% of the dried fruits.17 The LBPgroup has a molecular size range of 24–241 kDa, and severalLBPs have been isolated and purified from aqueousL. barbarum extracts by methods such as diethylaminoethylion-exchange cellulose and gel-permeation chromatogra-phy.14,18–20 Their structural composition has been studied bysodium dodecyl sulfate-polyacrylamide gel electrophoresis,gas chromatography, amino acid automatic analysis, partialacid hydrolysis, periodate oxidation, and nuclear magneticresonance spectrum, and they have been found to be com-plex glycopeptides consisting of acidic heteropolysac-charides and polypeptides or proteins. Although they differsomewhat in composition, the LBPs contain six monosac-charides (Ara, Rha, Xyl, Man, Gal, and Glc), galacturonicacid, and 18 amino acids. They also share a glycan-O-Ser

Manuscript received 28 October 2008. Revision accepted 13 December 2008.

Address correspondence to: Harunobu Amagase, Ph.D., Chief Scientific Officer, FreeLifeInternational, Inc., 4950 South 48th Street, Phoenix, AZ 85040, USA, E-mail: hamagase@freelife.com

JOURNAL OF MEDICINAL FOODJ Med Food 12 (5) 2009, 1159–1165# Mary Ann Liebert, Inc. and Korean Society of Food Science and NutritionDOI: 10.1089=jmf.2008.0300

1159

glycopeptide structure.20 The main chains of the glycanbackbones of LBPs have been found to be either a-(1? 6)-d-glucans or a-(1? 4)-d-polygalacturonans.21 LBPs havebeen focused upon as the active component responsible forvarious effects reported for L. barbarum. According toChinese understanding of Lycium extracts and products, thecontent of LBPs is important in the efficacy of L. barbarum.Other plant- and fungal-derived bioactive polysaccharideswith a broad range of immunomodulatory activities arefound in traditional Chinese medicine, and therefore a highcontent of polysaccharides with proven pharmacologicalactivities is considered to be an indicator of the medicinalstatus of the natural product.22,23

Along with an increasing number of studies of L. bar-barum, our recent randomized, double-blind, placebo-controlled clinical study showed that daily consumptionof L. barbarum for 14 days, provided in the form of afruit juice, GoChi� (FreeLife International LLC, Phoenix,AZ, USA), standardized for its LBP content, increasedsubjective feelings of general well-being, neurological=psychological assessment, and gastrointestinal function.1 Inthe present study, we have extended the clinical analysis ofGoChi as an individual herbal treatment to include measuresof immune function as well as its general subjective effectsand safety in older healthy human subjects.

MATERIALS AND METHODS

L. barbarum and placebo preparation

FreeLife International LLC supplied a commerciallyavailable, LBP-standardized L. barbarum fruit juice(GoChi; lot number ASA07120), which was produced fromfresh ripe L. barbarum fruit grown in Ningxia Province,China. Botanical identification was performed by JianguoLi, the director of the Ningxia Goji Institute (Yinchuan,China). The yield of juice as a percentage weight of thestarting fresh plant material is about 35%. Juice was pro-cessed in an aseptic manner and kept refrigerated before useat 2–88C. Description and standardization procedures for thetest material were previously described.1 In brief, GoChi isstandardized to contain a content of LBP equivalent to thatfound in at least 150 g of fresh fruit, the amount customarilyconsumed in traditional Chinese medicine.24

Placebo control material (lot number A198) was carefullyprepared to match the color, flavor, and taste of GoChi in aformulation of sucralose (10 mg), artificial fruit flavor(30 mg), citric acid (60 mg), and caramel color (12 mg) in30 mL of purified water. It was packaged in the same type ofcontainer; however, it provided no nutritional value or LBP.

Study population

All recruited subjects were healthy Chinese residents ofHunan Province, China. Subjects were men and womenbetween 55 and 72 years of age. All subjects had no obviousbrain, heart, liver, lung, kidney, or blood diseases, no historyof long-term medication, and no experience of GoChi in-take. Subjects were to discontinue use of any L. barbarum or

L. barbarum-containing foods at least 2 months before thetrial and throughout the study. A total of 60 participantswere selected for the study and were randomly assigned toeither the GoChi treatment group (n¼ 30; averageage¼ 58.70� 5.32 years) or placebo control (n¼ 30; aver-age age¼ 59.03� 5.62 years). All subjects were fully in-formed of the purpose of the study and signed the HumanSubjects Informed Consent forms approved under the Hel-sinki Declaration by the Internal Review Board of theHunan Provincial Center for Disease Control and Preventionin China. No participants were pregnant during the study.All subjects consumed their normal Chinese foods, and theiroriginal dietary habits were not changed during the trialperiod. Their normal diet included digestible foods, such asmilk, egg, soybean milk, noodles, gruel, vegetable, fruit, andothers, which constituted over 80% of the daily intake, aswell as small amount of deep-fried, greasy foods. Abdom-inal B-ultrasounds, electrocardiograms and chest X-rayexams of these subjects diagnosed right before the inter-vention trial were all within normal ranges. Mood and sleepparameters of the subjects were all normal.

Trial design

Subjects were randomly assigned to either the GoChi orplacebo groups. No dropouts were observed during the trial.All subjects were given a medical exam, and physical mea-surements (body weight, blood pressure, pulse rate, abdom-inal B-ultrasound, electrocardiogram, and chest X-ray exam)were assessed. Also, all subjects completed questionnairesand provided background information, including diseasehistory. Finally, 10 mL of venous blood was collected at pre-and post-intervention, allowed to clot, and centrifuged, andthe serum was stored at �708C for subsequent assays.

Subjects consumed 60 mL of the GoChi or placebo twicedaily (total, 120 mL=day) in the morning and evening after ameal every day for a period of 30 days. The dosage wasbased upon the quantity recommended in traditional Chi-nese medicine as previously described.1 All participantswere monitored daily to ensure full compliance with theprotocol. At the end of the 30-day intervention period,subjects were again given a medical exam, morphometricdata were recorded, the questionnaire was completed againby each participant, and venous blood samples were col-lected. Individuals administering the physical exam or ques-tionnaire were blinded as to the treatment conditions, andthe treatment codes were not broken until the study wascompleted. Randomization information was concealed fromthe investigators and the subjects until the end of the study.

Observation indicators

Each observation indicator was tested once upon com-mencement of the trial and at the end of the 30-day inter-vention period.

General physical exam. Questionnaire and physicalexam were used to assess the subjects’ energy, sleep, diet,

1160 AMAGASE ET AL.

urine, stool, body weight, blood pressure, and pulse ratecollected at pre- and post-intervention.

Urine panel. The urine panel tests were done with aurine analyzer (Shanghai Precison Instruments Co., Ltd.,Shanghai, China) for pH (5.5), leukocytes (�15 cells=mL),and urine glucose (mmol=L).

Stool panel. Stool test was determined by yellow colorindication, and its characterization was analyzed by strip; noblood clots, purulence, or helminths were safety indications.Microscopic examination was done, and criteria were ab-sence of erythrocytes or ovum and only a few plant cells andmuscle fibers.

Abdominal B-ultrasound, electrocardiogram, and chestX-ray exam. These tests were performed once immediatelybefore the trial. All subjects were within normal ranges.

Blood biochemical indicator tests. Routine analysis ofblood consisted of number of erythrocytes, number of leu-kocytes and their classification, and content of hemoglobinwith a full automatic hemocyte analyzer (Tianjin YixiangfaTechnology Co., Ltd., Hebei, China). Standard clinical testswere used to measure serum total protein, albumin, alaninetransaminase, aspartate transaminase, cholesterol, trigly-cerides, uric acid, blood urea nitrogen (BUN), creatinine,and blood glucose.

General and adverse reaction observations. Detailedquestions of the patient’s medical history were asked toassess the patient’s physical condition; clinical symptomswere observed and scored as follows: severe, 3 points;moderate, 2 points; mild, 1 point; none, 0 points. Dizziness,physical fatigue, and quality of sleep were used to evaluategeneral feelings of well-being.

Vision testing. Because we are interested in the efficacyof GoChi in visual functions based on the reports thatL. barbarum was effective in reducing the symptomsrelated to glaucoma and other eye diseases,25–27 vision wastested once at pre- and post-intervention. Subjects stood orsat 5 m from the international standard vision table and wereasked to cover one eye and then recognize the smallest letterthey could see on the table. Each eye was tested separately.

Short-term memory test. To begin, the subjects listenedto a recording of a native speaker reading a paragraph inChinese. Following the recording, the subjects took a 15-question test to see which words they remembered. Eachquestion had four choices. The subjects had to circle theword they had heard in the recording. Some of the questionshad four choices that sounded alike; in English, for example,weather, whether, wither, and wetter are four words thatsound alike. Some of the questions had four choices thathave the same meaning; in English, for example, method,way, manner, and system would be four words with the

same meaning. Some of them had four unrelated choices.For instance, weather, method, love, and result could beused as four unrelated words.

Focus testing. In every line of figures, there are somefigures with a neighbor, for which as a pair the two equal 10.Subjects were asked to focus attention on the line, find thesenumbers, and write the answer below the row. Every subjectshould be done within 7 minutes. An example is:

2946119355

Tests of immunological indicators. Venous blood sam-ples were collected at pre- and post-intervention to measurenumbers of CD4, CD8, and natural killer (NK) cells andlymphocytes and levels of interleukin (IL)-2, IL-4, immu-noglobulin (Ig)G, and IgA. A flow cytometer (Beijing Jun-HeHua Technology Co., Ltd., Beijing, China) was used tomeasure the number of CD4, CD8, and NK cells and lym-phocytes. Specific enzyme-linked immunosorbent assay kits(Zhengzhou AUTOBIO Co., Ltd. [Zhengzhou, China] andChengdu Shuguang Biotech Co., Ltd. [Chengdu, China])were used to measure the other immunological markers (IL-2, IL-4, IgG, and IgA levels) in serum.

Statistical analysis

Results were summarized as mean� SEM values. Forself-paired data t tests were used, and homogeneity of var-iance between the GoChi group and placebo group was aprerequisite. For mean comparisons group t tests were used.Otherwise, t tests were performed after homogeneity of var-iance was achieved through variable conversion. Rank-sumtests were used if the variance still lacked homogeneity. Anonparametric test was used to test for treatment effects onsleep and general feelings of well-being. The data wereprocessed using Statitica version 8 (StatSoft, Inc., Tulsa,OK, USA). Differences were considered significant atP< .05.

RESULTS

Safety observations

After consumption of GoChi or placebo for 30 days, noabnormalities were observed in subjects’ energy, sleep, diet,urine, stool, body weight, blood pressure, or pulse ratemeasures.

Parametric data

As shown in Tables 1 and 2, at pre- and post-intervention,there were minimal changes in body weight, blood pressure,pulse rate, urine, stool, blood panel, and blood biochemistryfor either the GoChi or the placebo control group. Allparametric data, such as body weights, etc., were analyzedby t test for independent and for dependent groups,and there were no significant changes in any of thesedependent measures between pre- and post-intervention

L. BARBARUM JUICE CLINICAL IMMUNOMODULATION 1161

for either group or any group differences, as shown inTable 2.

L. barbarum is well known in traditional Chinese herbalmedicine for treating diabetes, and this is supported byseveral reports showing an antidiabetic effect.25–30 Al-though we examined blood glucose levels in healthy oldersubjects, because of their general healthy condition andnormal blood glucose levels, no changes were seen in thepresent study.

LBPs have been shown to inhibit the increase of BUN andaccelerate its clearance after strenuous exercise,18 but in thepresent study GoChi did not influence BUN or other bio-chemical blood markers, possibly because of the absence ofa strenuous exercise challenge.

Observation of symptoms

As shown in Tables 3 and 4, GoChi treatment pro-duced significant changes (P< .05) in general feelingsof well-being over the trial period. Over 60% of subjectsin the GoChi group reported increased ratings of gen-eral well-being, which was statistically significantlydifferent from the placebo group. Only 20% of the sub-jects in the placebo group showed increased ratings to

these subjective questions, which were not statisticallysignificant.

Visual functions

Neither group showed any significant differences orchanges in their visual functions between pre- and post-intervention. L. barbarum has been reported to improve age-

Table 1. General Effects and Safety Aspects of GoChi or Placebo on Various Indications Pre- and Post-Intervention

Placebo (n¼ 30) GoChi (n¼ 30)

Pre-intervention Post-intervention Pre-intervention Post-intervention

Body weight (kg) 60.91� 1.74 61.02� 1.73 61.78� 1.77 61.78� 1.73Blood pressure (mm Hg)

Systolic 118.33� 2.33 118.07� 2.18 118.63� 2.44 118.90� 2.17Diastolic 73.60� 1.46 74.07� 1.25 74.70� 1.46 74.90� 1.17

Pulse (beats=minutes) 73.77� 1.21 74.13� 1.07 72.77� 1.04 73.03� 1.00White blood cells (109=L) 6.36� 0.28 6.39� 0.35 6.28� 0.35 6.45� 0.29Red blood cells (1012=L) 4.33� 0.07 4.35� 0.09 4.27� 0.08 4.30� 0.06Total protein (g=L) 134.90� 1.93 133.07� 1.76 133.07� 2.21 132.20� 1.82Urine Normal Normal Normal NormalStool Normal Normal Normal Normal

Data are mean� SEM values. Each indicator was tested once at pre- and post-intervention of the 30-day trial. Standard clinical tests and routine analysis of blood

were used.

Table 2. Biochemical Parameter Changes in Blood Pre- and Post-Intervention with GoChi or Placebo

Placebo (n¼ 30) GoChi (n¼ 30)

Pre-intervention Post-intervention Pre-intervention Post-intervention

Alanine transaminase (U=L) 23.63� 1.91 18.70� 1.81 23.60� 2.78 18.03� 1.78Aspartate transaminase (U=L) 26.47� 1.38 23.40� 1.33 25.87� 1.17 22.60� 0.83Cholesterol (mmol=L) 4.99� 0.14 4.87� 0.16 5.35� 0.17 5.26� 0.19Triglycerides (mmol=L) 1.41� 0.16 1.40� 0.20 1.66� 0.19 1.63� 0.17Total protein (g=L) 75.08� 1.03 70.65� 0.66 74.80� 0.85 69.56� 0.75Albumin (g=L) 46.70� 0.59 44.40� 0.45 46.14� 0.39 44.17� 0.45BUN (mmol=L) 4.55� 0.20 4.62� 0.22 5.00� 0.21 4.93� 0.21Creatinine (mmol=L) 75.07� 2.70 68.07� 3.56 77.57� 2.86 67.00� 3.81Uric acid (mmol=L) 257.37� 12.0 255.30� 9.80 245.23� 10.35 241.93� 12.36Blood glucose (mmol=L) 4.98� 0.09 5.02� 0.11 5.12� 0.07 4.95� 0.08

Data are mean� SEM values.

Table 3. Overall Clinical Symptoms After Consumption

of GoChi or Placebo

Group (n) Pre-intervention Post-intervention

Placebo (30) 4.27� 0.37 4.00� 0.37GoChi (30) 4.33� 0.31 3.00� 0.30ab

All subjects were administered a written questionnaire for which the

subjects provided a rating (scale of 0–3). The questionnaire consisted of

physical and psychological fatigue-related symptoms, such as fatigue,

dizziness, and sleep. These clinical symptoms were observed and scored as

follows: severe, 3 points; moderate, 2 points; mild, 1 point; none, 0 points.

Overall scores were obtained by summed each scores from symptoms and

statistically analyzed by a nonparametric test. Data are mean� SEM values.aSignificant difference compared to placebo, P< .05.bSignificant difference compared to pre-intervention, P< .05.

1162 AMAGASE ET AL.

related macular degenerative disease,25–27 but subjects inthe present study were all healthy and showed no symptomof age-related macular degenerative disease.

Short-term memory and attention capability

No statistically significant differences were found be-tween the placebo and GoChi groups, as shown in Table 5,but there was a tendency for increased short-term memoryand focus between pre- and post-intervention in the GoChigroup (P< .10).

Immunological marker changes in human serum

As shown in Table 6 and Figure 1, the GoChi-treatedgroup showed significant increases in number of lympho-cytes and levels of IL-2 and IgG, relative to pretreatmentlevels (P< .05). The percentage of CD4, CD8, and NK cellsand levels of IL-4 and IgA were not altered by GoChitreatment. None of the immune markers was changed fol-lowing the 30-day treatment period in the placebo controlgroup.

DISCUSSION

The present study is the first human clinical study re-porting immunomodulatory and other effects of L. barbar-um fruit in a form of juice (GoChi) as a single herb in healthyolder human subjects. Lymphocyte numbers and IL-2 and

Table 4. Effect of GoChi Compared to Placebo

on General Well-Being, Such as Dizziness, Fatigue,

and Sleep, and Total Indications

Number of subjects with positive response=totalnumber of subjects (% positive)

Placebo (n¼ 30) GoChi (n¼ 30)

Dizziness 5=24 (20.8%) 12=24 (50.0%)Fatigue 2=18 (11.1%) 11=21 (52.4%)Sleep 1=26 (3.9%) 8=25 (32.0%)Total 6=30 (20.0%) 19=30 (63.3%)a

Clinical symptoms were observed and scored as follows: severe, 3 points;

moderate, 2 points; mild, 1 point; none, 0 points. Dizziness, physical fatigue,

and quality of sleep were used to evaluate general feelings of well-being.aSignificant difference compared to placebo, P< .05.

Table 6. Immunological Changes After Consumption

of GoChi Compared to Placebo

Immunological marker,group (n) Pre-intervention Post-intervention

CD4 (%)Placebo (30) 53.21� 2.03 52.76� 1.80GoChi (30) 53.80� 1.86 54.95� 1.79

CD8 (%)Placebo (30) 34.54� 1.67 35.47� 1.89GoChi (30) 33.22� 2.09 36.59� 2.18

NK (%)Placebo (30) 0.67� 0.02 0.67� 0.03GoChi (30) 0.69� 0.03 0.73� 0.03

Lymphocytes (�106=mL)Placebo (30) 1.82� 0.03 2.09� 0.04GoChi (30) 1.76� 0.05 2.23� 0.03ab

IL-2 (pg=mL)Placebo (30) 6.09� 0.44 7.48� 0.54GoChi (30) 5.94� 0.53 9.36� 0.60ab

IL-4 (pg=mL)Placebo (30) 10.35� 0.66 11.42� 0.36GoChi (30) 10.79� 0.88 12.24� 0.78

IgG (g=L)Placebo (30) 14.70� 0.58 14.98� 0.63GoChi (30) 14.12� 0.52 16.78� 0.50ab

IgA (mg=L)Placebo (30) 2,586.33� 230.38 2,733.90� 191.66GoChi (30) 2,709.23� 188.50 3,280.87� 275.60

Venous serum samples were collected at pre- and post-intervention of the

trial. A flow cytometer was used to measure the number of CD4, CD8, and NK

cells and lymphocytes. Specific enzyme-linked immunosorbent assay kits were

used to measure the other immunological markers. Data are mean� SEM

values.aSignificant difference compared to placebo, P< .05.bSignificant difference compared to pre-intervention, P< .05.

Table 5. Effect of GoChi Compared to Placebo

on Short-Term Memory and Attention Capability

Task, group (n) Pre-intervention Post-intervention

Short-term memoryPlacebo (30) 2.47� 0.16 2.53� 0.14GoChi (30) 2.40� 0.12 2.67� 0.11b

AttentionPlacebo (30) 3.57� 0.31 3.70� 0.28GoChi (30) 3.43� 0.26 3.87� 0.20a

In the short-term memory test, the subjects first listened to a recording of a

native speaker reading a paragraph in Chinese. Following the recording, the

subjects took a 15-question test to see which words they remembered. In focus

attention testing, there are some figures in a line with a neighbor, for which as

a pair the two equal 10. Subjects were asked to focus attention on the line, find

these numbers, and write the answer below the row. Every subject should be

done within 7 minutes. Data are mean�SEM values.aTrend compared to pre-intervention, P< .10.

FIG. 1. Immune-stimulating effect of GoChi on several immunemarkers in human serum. Data are mean percentage changes from thepre-intervention value. **P< .01, *P< .05. Color images availableonline at www.liebertonline.com=jmf.

L. BARBARUM JUICE CLINICAL IMMUNOMODULATION 1163

IgG levels were significantly increased by GoChi intake.These study results suggest that GoChi may haveimmunomodulatory effects, which is consistent with tradi-tional usage and nonclinical studies of L. barbarum. Thesenonclinical studies have shown a wide variety of immuno-modulatory functions, including activation of various im-mune cells with enhancements of both cell-mediated andhumoral immune responses.11 Chemical analysis of LBPshas identified multiple-molecular-weight constitu-ents.14,18,20 These LBPs are structurally similar to com-pounds isolated from medicinal mushrooms and fromseveral herbs, including Astragalus, that have been shown tostimulate immune cells, have antioxidant properties, reducegastric irritation, and may protect against neurologicaldamage related to Alzheimer’s disease.2 One potentialmechanism for the immunomodulatory action for theselarge molecules is that they share structural similarities withcellular components of microorganisms and can modulateimmune cells via cell surface receptors. One clinical studyhas shown successful immunotherapy with the combinationof lymphokine-activated killer cell=IL-2 therapy and LBPconsumption in advanced cancer patients. Lymphokine-activated killer cell=IL-2 plus LBP treatment led to largerincreases in NK and lymphokine-activated killer cell activitythan lymphokine-activated killer cell=IL-2 without LBP.31

Although we did not observe an increase in the percentageof NK cells, NK cell activity was not assessed in the presentstudy. However, we did observe an increase in IL-2 levels inthe GoChi-treated group, as well as an increase in IgG levelsand the number of lymphocytes.

The antioxidant properties of GoChi may contribute to theobserved immune changes.32 Other dietary antioxidantshave been shown to produce an increase in IL-2 and IgGlevels as well as lymphocyte number.33–35 It is of interestthat the same immune measures that were changed withGoChi treatment in the present study have been reportedto show a decline with age.34,36,37 Given that the subjects inthe current study were older healthy adults (55–72 yearsold), the GoChi-induced increase in IL-2 and IgG levels andlymphocyte number may indicate an anti-aging effect on theimmune system. For example, the potentiation of IL-2production and T-cell function by dietary antioxidants hasbeen shown to be much more pronounced in older animals,relative to their younger controls.34,36

Additional changes found in the GoChi-treated group,relative to placebo controls, were significant reductions indizziness and fatigue and improvements of sleep withGoChi, results that are comparable to our previous study.1

The present study also found there was a tendency for in-creased short-term memory and focus. L. barbarum is re-garded as exhibiting anti-aging effects in traditional Asianmedicine as it has been shown to have effects on the brainand neuronal function associated with age-related neuro-degenerative diseases.24,38 Whether the antioxidant proper-ties represent a primary mechanism for the multiple effectsof GoChi requires additional investigation.

The present and our previous study1 confirmed the safetyof GoChi with daily consumption of 120 mL, equivalent to

at least 150 g of L. barbarum fresh fruit for 14–30 days inhumans. The safety of even high doses of GoChi has beenconfirmed in animal studies.39 There were two recent casereports of a possible interaction of L. barbarum fruit teawith warfarin,40,41 but the inhibition observed was weak,suggesting that the observed interaction may be caused byfactors other than the cytochrome P450 system. Further,given the high frequency of use of L. barbarum fruit and ofwarfarin, the lack of more reports of interaction suggeststhat the incidence may be very low.42 An independent labanalysis has shown that atropine has not been detected inGoChi, even though it is in a family of Solanacea. Thus,considering all the efficacy and safety information, GoChimay be safely taken without serious adverse effects.

In conclusion, a randomized, placebo-controlled, double-blind clinical study has shown daily consumption of LBP-standardized L. barbarum fruit juice, GoChi, for 30 dayssignificantly increased subjective feelings of general well-being and increased lymphocyte number and IgG and IL-2levels in Chinese normal healthy older human subjectswithout toxicity.

AUTHOR DISCLOSURE STATEMENT

All sources of support were from FreeLife International.H.A. is an employee of FreeLife International, and B.S. wassupported by FreeLife for pursuing clinical study in China.D.M.N. is a member of FreeLife’s Independent ScientificAdvisory Board.

REFERENCES

1. Amagase H, Nance DM: A randomized, double-blind, placebo-

controlled, clinical study of the general effects of a standardized

Lycium barbarum (Goji) juice, GoChi. J Altern Complement Med

2008;14:403–412.

2. Chang RC, So KF: Use of anti-aging herbal medicine, Lycium

barbarum, against aging-associated diseases. What do we know

so far? Cell Mol Neurobiol 2008;28:643–652.

3. Gruenwald J, Brendler T, Jaenicker C: Lycium berries (Go-Qi-Zi)

Lycium barbarum. In: PDR for Herbal Medicines, 4th ed.

Thompson Healthcare, Inc., Montvale, NJ, 2007, pp. 541–542.

4. Chang HM, But PPH: Gouqizi. In: Pharmacology and Applica-

tions of Chinese Materia Medica, Vol. 2. World Scientific, Sin-

gapore, 2001, pp. 852–854.

5. Bensky D, Gamble A: Gou Qi Zi. In: Chinese Herbal Medicine,

Materia Medica, revised ed. Eastland Press Inc., Seattle, 1993,

pp. 333–334.

6. Gan L, Hua Zhang S, Liang Yang X, Bi Xu H: Immuno-

modulation and antitumor activity by a polysaccharide-protein

complex from Lycium barbarum. Int Immunopharmacol 2004;4:

563–569.

7. Zhu J, Zhao LH, Zhao XP, Chen Z: Lycium barbarum polysac-

charides regulate phenotypic and functional maturation of murine

dendritic cells. Cell Biol Int 2007;31:615–619.

8. Luo Q, Yan J, Zhang S: [Effects of pure and crude Lycium

barbarum polysaccharides on immunopharmacology]. Zhong

Yao Cai 1999;22:246–249.

9. Gan L, Zhang SH, Liu Q, Xu HB: A polysaccharide-protein

complex from Lycium barbarum upregulates cytokine expression

1164 AMAGASE ET AL.

in human peripheral blood mononuclear cells. Eur J Pharmacol

2003;471:217–222.

10. Du S, Qian Y: Lycium barbarum extract’s effects on the ex-

pression of human interleukin-2 receptors (IL-2R). Chin J Mi-

crobiol Immunol 1995;15:176–178.

11. Wang BK, Xing ST, Zhou JH: Effect of Lycium barbarum

polysaccharides on the immune responses of T, CTL and NK

cells in normal and cyclophosphamide-treated mice. Chin J

Pharmacol Toxicol 1990;4:39–43.

12. Du G, Liu L, Fang J: Experimental study on the enhancement of

murine splenic lymphocyte proliferation by Lycium barbarum

glycopeptide. J Huazhong Univ Sci Technolog Med Sci 2004;

24:518–520, 527.

13. Yin J, Guo LG: Regulatory action of licorice and Lycium fruit on

IgE antibodies response. J Beijing Med Univ 1992;24:115–117.

14. Peng X, Tian G: Structural characterization of the glycan part of

glycoconjugate LbGp2 from Lycium barbarum L. Carbohydr Res

2001;331:95–99.

15. Zhu YP: Gou Qi Zi. In: Chinese Materia Medica: Chemistry,

Pharmacology and Applications. Harwood Academic Publishers,

Amsterdam, 1998, pp. 642–646.

16. National Health Information, NHIOnDemand, LLC: Gou Qi

Zi, Lycium Fruit, Lycii Fructus. http:==content.nhiondemand

.com=moh=media=TCMH1.asp?objID¼100832&ctype¼tcmh#fn

111585 (accessed November 2007).

17. Wang Q: Determination of polysaccharide contents in fructus

Lycii. Chin Tradit Herb Drugs 1991;22:67–68.

18. Luo Q, Yan J, Zhang S: [Isolation and purification of Lycium

barbarum polysaccharides and its antifatigue effect]. Wei Sheng

Yan Jiu 2000;29:115–117.

19. Peng XM, Wang ZF, Tian GY: [Physico-chemical properties and

activity of glycoconjugate LbGp2 from Lycium barbarum L.].

Yao Xue Xue Bao 2001;36:599–602.

20. Tian M, Wang M: [Studies on extraction, isolation and compo-

sition of Lycium barbarum polysaccharides]. Zhongguo Zhong

Yao Za Zhi 2006;31:1603–1607.

21. Duan CL, Qiao SY, Wang NL, Zhao YM, Qi CH, Yao XS:

[Studies on the active polysaccharides from Lycium barbarum

L.]. Yao Xue Xue Bao 2001;36:196–199.

22. Chang R: Bioactive polysaccharides from traditional Chinese

medicine herbs as anticancer adjuvants. J Altern Complement

Med 2002;8:559–565.

23. Wong CK, Leung KN, Fung KP, Choy YM: Immunomodulatory

and anti-tumour polysaccharides from medicinal plants. J Int

Med Res 1994;22:299–312.

24. Yu MS, Lai CS, Ho YS, Zee SY, So KF, Yuen WH, Chang RC:

Characterization of the effects of anti-aging medicine Fructus

lycii on beta-amyloid peptide neurotoxicity. Int J Mol Med

2007;20:261–268.

25. Li XM: Protective effect of Lycium barbarum polysaccharides on

streptozotocin-induced oxidative stress in rats. Int J Biol Mac-

romol 2007;40:461–465.

26. He J, Zhou W, Qiu Y: A clinical study in the effect of Lycium

barbarum L. on antioxidizing reaction in the patients with

diabetic retinopathy. J Chin Tradit Med Ophthalmol 1998;8:

74–76.

27. He J, Zhou W, Qiu Y: The changes in immunologic functions of

red blood cell in the patients with diabetic retinopathy and the

effect of Lycium barbarum L. in the treatment. J Chin Tradit Med

Ophthalmol 1998;8:70–73.

28. Wang L, Zhang C, Li W: Regulating effect of Lycium barbarum

polysaccharide on T lymphocyte subpopulation and cytokines in

patients with type-2 diabetes. Hebei J Tradit Chin Med 2001;

23:888–890.

29. Li W, Wang L, Deng X, Jiang L, Zhang CJ, Zhang C: Study of

the fragility and abnormality rate of red blood cells in patients

with type-2 diabetes and the effects of Lycium barbarum poly-

saccharides. Hebei J Tradit Chin Med 2000;22:585–586.

30. Wu H, Guo H, Zhao R: [Effect of Lycium barbarum polysac-

charide on the improvement of antioxidant ability and DNA

damage in NIDDM rats]. Yakugaku Zasshi 2006;126:365–371.

31. Cao GW, Yang WG, Du P: [Observation of the effects of

LAK=IL-2 therapy combining with Lycium barbarum polysac-

charides in the treatment of 75 cancer patients]. Zhonghua Zhong

Liu Za Zhi 1994;16:428–431.

32. Amagase H, Sun B, Borek C: Lycium barbarum (goji) juice

improves in vivo antioxidant biomarkers in serum of healthy

adults. Nutr Res 2009;29:19–25.

33. Hughes DA: Dietary antioxidants and human immune function.

Br Nutr Found Nutr Bull 2000;25:35–41.

34. Adolfsson O, Huber BT, Meydani SN: Vitamin E-enhanced IL-2

production in old mice: naive but not memory T cells show

increased cell division cycling and IL-2 producing capacity.

J Immunol 2001;167:3089–3817.

35. Al-Ani FS, Al-Samaraee IF, Abdul-Razzak AR: Antioxidants

prolong survival of lymphocytes. Med J Islamic Acad Sci 2000;

13:141–145.

36. Han SN, Meydani SN: Antioxidants, cytokines, and influenza

infection in aged mice and elderly humans. J Infect Dis 2000;

182(Suppl 1):S74–S80.

37. Goidl EA, Innes JB, Weksler ME: Immunological studies of

aging. II. Loss of IgG and high avidity plaque-forming cells and

increased suppressor cell activity in aging mice. J Exp Med 1976;

144:1037–1048.

38. Yu MS, Ho YS, So KF, Yuen WH, Chang RC: Cytoprotective

effects of Lycium barbarum against reducing stress on endo-

plasmic reticulum. Int J Mol Med 2006;17:1157–1161.

39. Amagase H: General toxicity and histological analysis from acute

toxicological study of a standardized Lycium barbarum (goji)

juice (GoChi�) in rodents [abstract]. FASEB J 2008;22:lb722.

40. Lam AY, Elmer GW, Mohutsky MA: Possible interaction be-

tween warfarin and Lycium barbarum. Ann Pharmacother 2001;

35:1199–1201.

41. Leung H, Hung A, Hui ACF, Chan TYK: Warfarin overdose due

to the possible effects of Lycium barbarum L. Food Chem Tox-

icol 2008;46:1860–1862.

42. Dharmananda S: Lycium Fruit, Food and Medicine. http:==www

.itmonline.org=arts=lycium.htm (accessed January 2008).

L. BARBARUM JUICE CLINICAL IMMUNOMODULATION 1165