Investigating peripheral blood monocyte and T-cell subsets as non-invasive biomarkers for asymptomatic hepatic steatosis: results from the Multi-Ethnic Study of Atherosclerosis.

Background: Circulating immune cells have gained interest as biomarkers of hepatic steatosis. Data on the relationships between immune cell subsets and early-stage steatosis in population-based cohorts are limited. Methods: This study included 1,944 asymptomatic participants of the Multi-Ethnic Study of Atherosclerosis (MESA) with immune cell phenotyping and computed tomography measures of liver fat. Participants with heavy alcohol use were excluded. A liver-to-spleen ratio Hounsfield units (HU) <1.0 and liver attenuation <40 HU were used to diagnose liver fat presence and >30% liver fat content, respectively. Logistic regression estimated cross-sectional associations of immune cell subsets with liver fat parameters adjusted for risk factors. We hypothesized that higher proportions of non-classical monocytes, Th1, Th17, and memory CD4+ T cells, and lower proportions of classical monocytes and naive CD4+ T cells, were associated with liver fat. Exploratory analyses evaluated additional immune cell phenotypes (n = 19). Results: None of the hypothesized cells were associated with presence of liver fat. Higher memory CD4+ T cells were associated with >30% liver fat content, but this was not significant after correction for multiple hypothesis testing (odds ratio (OR): 1.31, 95% confidence interval (CI): 1.03, 1.66). In exploratory analyses unadjusted for multiple testing, higher proportions of CD8+CD57+ T cells were associated with liver fat presence (OR: 1.21, 95% CI: 1.02, 1.44) and >30% liver fat content (OR: 1.34, 95% CI: 1.07, 1.69). Conclusions: Higher circulating memory CD4+ T cells may reflect liver fat severity. CD8+CD57+ cells were associated with liver fat presence and severity, but replication of findings is required.

Enlargement of the Spleen Index Is a Predictor of the Occurrence of Esophageal Varices and Hepatocellular Carcinoma after Administering Direct-acting Antiviral Agents.

Objective Direct-acting antiviral agents (DAAs) can eliminate hepatitis C virus at a high rate, although the long-term incidence of portal hypertension and hepatocellular carcinoma (HCC) has not yet been elucidated. In this observational study, we clarified the predictors associated with the incidence of esophageal varices (EVs) and HCC after DAAs treatment based on ultrasound findings and blood examinations. Methods A total of 78 patients treated with DAAs were enrolled in this study. The primary endpoint was to identify the predictors associated with EVs and HCC occurrence using univariate and multivariate analyses. Secondary endpoints were to extract the cutoff values for EVs and HCC occurrence and clarify the changes in liver stiffness (LS), spleen stiffness (SS), spleen index (SI), portal venous flow volume (PVF), and blood examination at 12 weeks after the end of DAAs treatment. Results The mean observation period was 1,402±546 days. SI change (SI after DAAs-SI before DAAs) was a predictor of EVs occurrence in multivariate analysis (p=0.045). The treatment history of HCC, albumin value before DAAs, and SI change were predictors of HCC occurrence in multivariate analysis (p=0.002, p=0.032, and p=0.009, respectively). LS, SS, PVF, SI, and liver function significantly improved after DAAs treatment. Conclusion Portal hypertension seems to improve after DAAs treatment over a long period. Patients with splenomegaly deterioration after DAAs treatment need to be carefully monitored for the occurrence of EVs and HCC.

An experimental study of buckwheat polysaccharide in adjuvant therapy for S180 sarcoma mice.

Introduction: To discuss the auxiliary therapeutic effect of buckwheat polysaccharide (BP) on S180 sarcoma. Material and methods: Buckwheat polysaccharide was extracted with water and precipitated with ethanol. Solid tumor and ascites tumor mice models were established. The mice in the high, medium and low dosing groups (n = 24, each group) had their stomachs filled with different doses of BP. The cyclophosphamide (CTX) group and the model group (n = 24, each group) were used as control groups. The influence on the life cycle, the rate of suppressing the tumor, the thymus index, and the spleen index were evaluated. Tumor cells were cultured in vitro and intervened with drugs; flow cytometry was used to detect the changes in the cell cycle. Results: Buckwheat polysaccharide significantly improved the lifespan and survival rate of the mice. The group of mice treated with the medium dose showed the best survival rate when compared to the ones that received high and low doses of BP (p < 0.01). The tumor cells cultured in vitro were arrested in the G0/G1 phase to some extent (p > 0.05). The cyclophosphamide arrested the cycle of the tumor cells in the G2/M period (p < 0.01). Buckwheat polysaccharide could increase the thymus index, spleen index and the rate of suppressing the tumor, but the differences were not statistically significant. Conclusions: Buckwheat polysaccharide had no obvious effect in inhibiting the growth of tumors, but it significantly extended the lifespan, increased the survival rate and reduced the toxic effect of CTX.

Measurement of multiple spleen lengths is not necessary for non-invasive prediction of high-risk esophagogastric varices.

AIM: To validate an appropriate spleen size measurement technique for the prediction of high-risk esophagogastric varices. METHODS: This retrospective cross-sectional study included 369 patients who underwent ultrasonography and computed tomography (CT) of the spleen and esophagogastroduodenoscopy between January 2018 and December 2020. Maximum spleen length, width, and craniocaudal length were measured in a longitudinal view. The two-dimensional (2D) spleen index (maximum length × maximum width in the longitudinal view) was calculated. A three-dimensional (3D) spleen index was then defined as follows: 2D spleen index × maximum length in the transverse view. The similarity in spleen volume measured by CT and ultrasonography (spleen index) was assessed by the correlation coefficient. The diagnostic accuracies of the spleen index, platelet/spleen length, and platelet/spleen index were calculated to determine the overall diagnostic accuracy. RESULTS: Compared to the other spleen indices, our 3D spleen index was significantly better correlated with spleen volume on CT (r = 0.91, 95% confidence interval 0.89-0.92, p < 0.001). Receiver-operating characteristic curve analyses revealed no significant difference between the 3D and 2D indices (p = 0.228) but did show a significant difference between the 3D and one-dimensional indices (p = 0.020). Although the area under the curve for the platelet count combined with the spleen index or length was higher than that for our 3D index, there was no significant difference between platelet count and spleen index or length (p = 0.078). CONCLUSIONS: Platelet/spleen length has a reasonable ability to predict high-risk esophagogastric varices, even though measurement of two or three factors can be correlated with spleen volume.

[Effect of electroacupuncture at "Zusanli" (ST 36) and its matching point at abdomen on intestinal motility in mice with postoperative ileus].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) at different acupoint combination on intestinal motility and expression of inflammatory factors IL-6 and TNF-α in intestinal muscle layer in mice with postoperative ileus (POI), so as to provide reference of acupoint combination of clinical EA intervention for POI. METHODS: A total of 78 SPF-grade male C57BL/6 mice aged 8-12 weeks were randomly divided into a sham operation group, a model group, a ST 36+ non-acupoint group, a ST 36+ ST 25 group, a ST 36+ CV 4 group and a ST 36+ CV 12 group, 13 mice in each group. The model of POI was established by intestinal manipulation. The mice in each acupuncture group were intervened with HANS-200A acupoint nerve stimulator within 3 hours after modeling, continuous wave, frequency of 10 Hz, intensity of 1 mA, for 20 min. The EA was given once. 22.5 h after model establishment, 7 mice in each group were intragastrically administered with fluorescein isothiocyanate-conjugated (FITC-dextran). 1.5 h after intragastric administration, intestinal motility was evaluated. 24 h after modeling, the expression of IL-6 and TNF-α mRNA in intestinal muscle layer was detected by real-time fluorescence quantitative PCR and spleen index was calculated. RESULTS: Compared with the sham operation group, the intestinal motility 24 h after modeling in the model group was significantly decreased (P<0.01), and the expression of IL-6 and TNF-α mRNA in intestinal muscle layer was increased (P<0.01). Compared with the model group, the intestinal motility 24 h after modeling in the ST 36+ non-acupoint group and the ST 36+ ST 25 group was increased (P<0.01), and the expression of IL-6 and TNF-α mRNA in intestinal muscle layer was decreased (P<0.01, P<0.05), and the expression of TNF-α mRNA in intestinal muscle layer in the ST 36+ CV 4 group was also decreased (P<0.05), but there was no significant difference in the ST 36+ CV 12 group and the ST 36+ CV 4 group (P>0.05). There was no significant difference in spleen index among the groups (P>0.05). CONCLUSION: EA at "Zusanli" (ST 36) or "Zusanli" (ST 36) combined with "Tianshu" (ST 25) can improve intestinal motility in mice with POI, and the mechanism may be related to the improvement of local inflammatory reaction. In addition, abdominal acupoints should be selected carefully.

Immunomodulatory influences of sialylated lactuloses in mice.

The aim of this study was to investigate the immune modulatory influences of sialylated lactuloses in mice. The effects of the four sialylated lactuloses by gavage methods on the weight gain rate, organ, serum and spleen immunoglobulin of mice were investigated. Neu5Ac-α2,3-lactulose group and Kdn-α2,3-lactulose group had significantly higher weight gain rate than control group. The weight gain rate, thymus index and spleen index of Kdn-α2,3-lactulose group were significantly higher than control group and lactulose group. Liver and small intestine of Neu5Ac-α2,3-lactulose group, Neu5Ac-α2,6-lactulose group and Kdn-α2,6-lactulose group showed different degree of damage. IgG levels of serum and spleen in Neu5Ac-α2,6-lactulose group and Kdn-α2,6-lactulose group were significantly higher than control group and lactulose group. The contents of IgG in serum and spleen of Kdn-α2,3-lactulose group were significantly lower than that of control group, while the contents of IgA and IgM in serum were significantly higher than those of control group. The IgA level increased by 12.23% and 58.77% comparing with lactulose group and control group, respectively. The IgM level in serum of Kdn-α2,3-lactulose group mice increased by 43.88% and 8.05% comparing with control group and lactulose group, respectively. The IgA level and IgM level in spleen of Kdn-α2,3-lactulose group mice increased by 49.05% and 47.25% comparing with control group. In short, Kdn-α2,3-lactulose is relatively safe and superior to use as a food supplement or potential drug candidate. Our results also indicate that some other sialylated oligosaccharides are potentially harmful to organisms, they may cause some side effects.

1-Deoxynojirimycin from Bacillus subtilis improves antioxidant and antibacterial activities of juvenile Yoshitomi tilapia

Background: Juvenile Yoshitomi tilapia is often infected by pathogens and results in low-level survival rate. Bacillus subtilis, as a probiotic, may have beneficial effects on Y. tilapia with compound 1-deoxynojirimycin (DNJ), which has antibacterial activities. The effects of dietary probiotic supplementation on Y. tilapias were evaluated. Results: Juvenile Y. tilapia was fed with B. subtilis for 56 d. Y. tilapia was infected by Aeromonas hydrophila and survival rate was compared. Dietary B. subtilis increased weight gain rate, specific growth, food conversion ratios and food intake rate of Y. tilapia. The diet improved the cumulative survival rate (CSR) of juvenile Y. tilapia when the concentration of B. subtilis was more than 2.05 × 1010 cfu/kg and CSR reached a maximum rate when the concentration of bacillus was 4.23 × 1010 (P b 0.05). Meanwhile, B. subtilis improved total antioxidant capacity (TAC), spleen index, the activities of serum lysozyme, alkaline phosphatase (ALP), superoxide dismutase (SOD) and catalase (CAT) (P b 0.05). In contrast, B. subtilis reduced serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA) and C3 complement (P b 0.05). DNJ was isolated from secondary metabolisms and proved to increase the levels of SOD, CAT and reduce the levels of AST, ALT and MDA at cell levels. After A. hydrophila infection, DNJ prevented the reduction in survival rate of Y. tilapia (P b 0.05). Conclusions: 1-Deoxynojirimycin from Bacillus subtilis can be used to improve the growth performance of juvenile Y. tilapia by affecting its antioxidant and antibacterial activities.

Cell-free DNA-induced alteration of autophagy response and TLR9-signaling: Their relation to amelioration of DSS-colitis.

BACKGROUND: The influence of cell-free DNA (fDNA) administration on the TLR9-autophagy regulatory crosstalk within inflammatory circumstances remains unclear. AIMS: To examine the immunobiologic effects of iv. fDNA injection on the TLR9-mediated autophagy response in murine DSS-colitis. METHODS: Different types of modified fDNAs were administered to DSS-colitic mice. Disease and histological activities, spleen index were measured. Changes of the TLR9-associated and autophagy-related gene expression profiles of lamina proprial cells and splenocytes were assayed by quantitative real-time PCR, and validated by immunohistochemistries. Ultrastructural changes of the colon were examined by transmission electron microscopy (TEM). RESULTS: A single intravenous injection of colitic fDNA (C-DNA) exhibited beneficial clinical and histological effects on DSS-colitis, compared to normal (N-DNA). C-DNA administration displayed a more prominent impact on the outcome of the TLR9-autophagy response than N-DNA. C-DNA resulted in a decreased spleen index in DSS-colitic mice. C-DNA treatment of normal mice resulted in a downregulation of Beclin1 and ATG16L1 mRNA and protein expression in the colon. These as well as LC3B were downregulated in the spleen. In contrast, the Beclin1, ATG16L1 and LC3B gene and protein expressions were upregulated in both the colon and the spleen by C-DNA injection. Moreover, C-DNA administration to DSS-colitic mice resulted in a remarkable increase of epithelial autophagic vacuoles representing an intensified macroautophagy. CONCLUSIONS: The effect of intravenously administered fDNA on the TLR9-mediated autophagy response is expressly dependent on the origin of fDNA (i.e. inflammatory or not) and on the characteristics of the local immunobiologic milieu (i.e. inflammatory or not, as well).

Correlación del índice plaqueta/ bazo con el grado de várices esofágicas / Correlation between platelet/spleen index and esophageal varices degree

Resumen ANTECEDENTES: las enfermedades hepáticas crónicas en México fueron motivo de 28,904 fallecimientos durante 2012. Su principal complicación fue la hipertensión portal, manifestada por várices esofágicas (VE) que afectan a 30% de los pacientes. OBJETIVO: utilizar la correlación del índice plaqueta/bazo (IPB) con el grado de várices esofágicas como método diagnóstico no invasivo y de decisión terapéutica. MATERIAL Y MÉTODO: estudio observacional, analítico, transversal y retrospectivo, en el que se recolectaron datos de pacientes con diagnóstico de insuficiencia hepática, atendidos en el servicio de Medicina Interna del Hospital Central Norte de Petróleos Mexicanos, entre enero de 2010 y abril de 2016; se calcularon puntos de corte para conocer el grado de asociación del IPB con el grado de várices esofágicas; el patrón de referencia fue la endoscopia digestiva alta. RESULTADOS: se encontró un nivel de significación con valor p de 0.005 y Rho de Spearman de -0.425 que traduce una correlación estadísticamente significativa entre las variables en correlación con la escala de Dagradi y nivel de significación con valor p de 0.001 y Rho de Spearman de -0.492 en correlación con la escala de Baveno. Asimismo, para la escala de Baveno un IPB mayor a 0.700 se correlacionó con várices esofágicas pequeñas, en tanto que un IPB menor a 0.700 se correlacionó con várices esofágicas grandes. CONCLUSIONES: esta correlación puede ser un método no invasivo útil que permite establecer prioridad en cuanto al abordaje diagnóstico y discernir entre iniciar tratamiento farmacológico o realizar un procedimiento endoscópico.

Abstract BACKGROUND: Chronic liver diseases in Mexico were cause of 28,904 deaths in 2012. Its main complication was portal hypertension, manifested by esophageal varices (EV) occurring in 30% of patients. OBJECTIVE: To use the correlation of platelet/spleen index (PSI) with the degree of EV as a noninvasive diagnostic method and therapeutic decision. MATERIAL AND METHOD: An observational, analytical, cross-sectional and retrospective study was done in which data were collected from patients with chronic liver failure assisted at the Department of Internal Medicine of North Central Hospital Pemex, Mexico City, from January 2010 to April 2016. Cutpoints were calculated to determine the degree of association between PSI with the degree of EV; using the upper gastrointestinal endoscopy as the gold standard. RESULTS: We found a significance level with p-value of 0.005 and -0.425 Spearman Rho, defaming a statistically significant correlation between the variables in correlation to Dagradi Scale and a level of significance with p-value of 0.001 and -0.492 Spearman Rho in correlation to Baveno Scale. For Baveno scale it was observed that a PSI higher than 0.700 correlated with small EV, while a PSI lesser than 0.700 PSI correlated with large EV. CONCLUSIONS: This correlation can be a non-invasive useful method which allows to set priority in regard to diagnostic approach and to dis cern between starting drug treatment or doing endoscopic procedure.

Characterization of basic immune function parameters in the fathead minnow (Pimephales promelas), a common model in environmental toxicity testing.

The fathead minnow (Pimephales promelas) is an environmental sentinel species, commonly used in toxicity testing. However, there is a lack of data regarding basic immune function in this species. To improve the usefulness of the fathead minnow as a model for basic immune function and immunotoxicity, this study sought to 1) compare the differential expression of immune function genes in naïve fathead minnows and 2) determine the effects of pathogen exposure on immune gene expression and spleen index. To accomplish this, kidney, spleen and liver tissue were collected three days post injection (dpi) from adult male fathead minnows from each of the following groups: 1) uninjected control 2) sham-injected (Hank's balanced salt solution) and 3) pathogen-injected (Yersinia ruckeri). Spleen tissue was also collected at seven and 14 dpi. Differential tissue expression of immune function genes was evaluated in naïve minnows and expression patterns were similar to those found in other fish species, with liver tissue generally having the highest amount of expression. Following pathogen injection, the expression of complement component 3 (c3) (4.4-fold, kidney; 2.5-fold, liver), interleukin 11 (il11) (4.8-fold, kidney; 15.2-fold, liver) and interleukin 1ß (il1ß) (8.2-fold, kidney; 17.2-fold, spleen; 2.6-fold, liver) were significantly upregulated. Elastase 2 (elas2) was significantly downregulated (5.8-fold) in liver tissue. A significant increase in spleen index at seven dpi was also observed in pathogen-injected minnows. This study has identified endpoints that are part of the normal response to pathogen in fathead minnows, an essential step toward the development of the fathead minnow as a model for immunotoxicity evaluations.

Nonfluorodeoxyglucose-Avid Persistent Splenomegaly at Time of Transplantation Delays Neutrophil and Platelets Engraftment without Affecting Survival in Patients with Lymphomas Undergoing Allogeneic Hematopoietic Cell Transplantation.

It is unclear if persistent splenomegaly in the presence of a negative positron emission tomography (PET) scan before allogeneic hematopoietic cell transplantation (HCT) influences post-transplantation outcomes in patients with lymphoma. We retrospectively reviewed records of 152 patients who underwent allogeneic HCT for various lymphomas. Centralized review of pretransplantation computed tomography (CT) and PET images was performed. Spleen volume (SV) was measured using the freehand volume segmentation tool in AW Workstation software (General Electric, Waukesha, WI). Splenic index (SI) was calculated as a product of width, thickness, and length of the spleen. Normal SV was defined as SV < 314.5 cm3 and normal SI was defined as SI ≤ 480 cm3, as described in the literature. Among the study population, 42.8% received an allogeneic HCT from an HLA-matched related donor, 36.2% from a matched unrelated donor, 12.5% from a mismatched unrelated donor, and 8.6% received a double umbilical cord blood transplantation. Most (61.8%) received myeloablative conditioning. Median age at transplantation was 52 (range, 21 to 68) years. Pre-allogeneic HCT spleen CT and PET images were available on 88% and 70.3% patients, respectively. SV ranged from 90 cm3 to 4684 cm3 with a median of 290.5 cm3 and a mean of 400.3 cm3. SI calculation showed a range from 50.3 cm3 to 8276.4 cm3 with a median of 582.1 cm3 and a mean of 771.2 cm3. The majority of patients (83.1%) had PET-negative spleen before allogeneic transplantation. Engraftment was delayed in PET-negative patients with persistent splenomegaly, with median days to neutrophil engraftment of 17 versus 16 (P = .03) and median days to platelet engraftment of 16 versus 14 (P = .04) when using SV. However, persistent splenomegaly did not appear to impact progression-free survival (P = .11) or overall survival (P = .37). Splenomegaly in the setting of a PET-negative study before allogeneic HCT delays neutrophil and platelet engraftment but does not appear to affect survival. Future studies using registry data or larger multicenter studies would be required to evaluate the impact of splenomegaly and its fluorodeoxyglucose avidity on allogeneic HCT outcomes in specific subtypes of lymphomas.

Shen-Qi-Jie-Yu-Fang has antidepressant effects in a rodent model of postpartum depression by regulating the immune organs and subsets of T lymphocytes.

BACKGROUND: Shen-Qi-Jie-Yu-Fang (SJ Fang) is a herbal preparation used in traditional Chinese medicine, and is a potentially important new therapeutic agent in postpartum depression (PPD). Previously, we have elucidated the effects of SJ Fang on hormone receptors and monoamine neurotransmitters involved in the hypothalamic-pituitary-adrenal and hypothalamic-pituitary-gonadal axes in PPD rats. However, the immune-modulating effects of SJ Fang in PPD are still unknown. In this study, we explored the effects of SJ Fang on the immune organs and subsets of T lymphocytes in PPD rats. METHODS: PPD was created in Sprague-Dawley rats by inducing hormone-simulated pregnancy followed by hormone withdrawal. After hormone withdrawal, the PPD rats were then treated with fluoxetine at 1, 2, and 4 weeks, and the SJ Fang rats were also treated at 1, 2, and 4 weeks. Depressive behavior in the rats was evaluated by the forced swim test, sucrose consumption test, and open field test. The thymus index and spleen index were calculated. Hematoxylin-eosin staining was used to identify pathological features in the thymus and spleen. CD3, CD4, and CD8 lymphocyte subsets were analyzed by flow cytometry. RESULTS: Both fluoxetine and SJ Fang decreased immobility time, increased sucrose consumption, an horizontal and vertical movement. After 4 weeks of treatment with fluoxetine or SJ Fang, the thymus index and spleen index were significantly higher than at baseline, and the morphology of the thymus and spleen were returning to normal. Two weeks after hormone withdrawal, subsets of T lymphocytes indicated a shift from immune activation to immune suppression, which was reversed by 4 weeks of treatment with fluoxetine or SJ Fang. CONCLUSION: It is suggested that T-cell mediate immune responses which may play a role in the etiopathology of postpartum depression. SJ Fang had an antidepressant effect on the immune system in rats with PPD.

Effects of Hypericum perforatum extract on IgG titer, leukocytes subset and spleen index in rats.

OBJECTIVES: Hypericum perforatum L. is a medicinal plant containing many polyphenolic compounds, including flavonoids and phenolic acids with antidepressant and anti-inflammatory properties. This study was investigated the effects of Hypericum perforatum extract (HPE) on immunity, body weight (BW), and spleen index (SI) in rats. MATERIALS AND METHODS: A total of 24 Wistar male rats were randomly received 4 different doses (6 rats each) of HPE (0, 100, 200 and 400 mg/kg BW) intraperitoneally for 14 days using a completely randomized design. On days 1 and 7, rats were received 0.5 ml SRBC (10%) injection. Blood samples were collected on day 14 to evaluate IgG titer and leukocyte count. On days 1, 7 and 14, the BW and on day 14 spleen were weighted for SI. RESULTS: The IgG titer increased with higher doses of HPE. The HPE increased number of lymphocytes at 200 mg but decreased at 400 mg, number of neutrophils decreased at 200 mg but increased at 400 mg, and number of monocytes increased at 100 mg and 200 mg but decreased at 400 mg (p<0.01). Increasing doses of HPE lowered BW (p<0.01). The HPE increased SI at 100 mg and 200 mg but decreased at 400 mg (p>0.072). CONCLUSIONS: The results showed that HPE slightly improved IgG titer but significantly increased the number of leukocytes and monocytes at 200 mg, and neutrophils at 400 mg. The HPE decreased BW at 100 mg and 200 mg with no damage on spleen.

Beneficial effects of 20(S)-protopanaxadiol on antitumor activity and toxicity of cyclophosphamide in tumor-bearing mice.

20(S)-protopanaxadiol (PPD) is an extract of Panax quinquefolius L. The aim of this study was to investigate the effect of PPD on the antitumor activity and toxicity of cyclophosphamide (CTX) in tumor-bearing mice. C57BL/6 mice bearing Lewis lung carcinoma cells were treated with PPD (50 mg/kg) alone, CTX (20 mg/kg) alone or PPD (50 mg/kg) in combination with CTX (20 mg/kg), respectively. The results showed that PPD alone has no significant antitumor activity but synergistically enhanced the antitumor activity of CTX. PPD significantly increased the peripheral white blood cell count, bone marrow cell count, interleukin-2 and interferon-γ in CTX-treated tumor-bearing mice. The lowered levels of spleen index, splenocyte proliferation and natural killer cell activity in tumor-bearing mice following CTX treatment were also increased by PPD administration. PPD may be a beneficial supplement during CTX chemotherapy for enhancing the antitumor efficacy and reducing the toxicity of CTX.