Er-Xian decoction exhibits protective activity in a rat model of asthenospermia through a mechanism associated with DJ-1 protein upregulation.

Asthenozoospermia (AZS) remains a significant clinical problem of male factor infertility. Er-Xian decoction (EXD) is a traditional Chinese medicine with potent antioxidant activity to treat AZS. To investigate the protective effects of EXD on sperm motility and deglycase (DJ)-1 expression in AZS model rats. Sixty mature male Sprague-Dawley rats (200 - 250 g) were randomized into five equally sized groups, including ornidazole (ORN)-induced AZS model group, or L-carnitine (0.1 g/kg) treated group or EXD group (7.5, 15, or 30 g crude drug/kg). Oxidative stress was assessed by measuring superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px). DJ-1 expression in testis and epididymis tissue was measured via qRT-PCR, Western blotting, and immunofluorescence staining. Hematoxylin and eosin staining was used to gauge morphological changes of testis and epididymis. Sperm motility was significantly reduced the AZS model group, while increased in the low-, intermediate-, and high-dose EXD treatment groups by 45.51%, 49.43%, and 58.31%, respectively (P < 0.001), which with a similar increase of 57.21% being observed in the L-carnitine treatment group. Relative to the control group, oxidative stress indices were significantly altered in AZS model rats, which exhibited significant reductions in SOD and GSH-Px levels and significantly increased MDA levels (49.44 ± 1.38 U/ml, 14.02 ± 0.70 U/ml, and 26.37 ± 1.03 nmol/ml, respectively). After EXD treatment, oxidative stress indexes were significantly improved relative to those in these model rats, with high-dose EXD yielding more significant improvements in these oxidative stress indices relative to L-carnitine treatment. While AZS model rats exhibited morphological abnormalities, tissue disorder, and reduced cell counts in the testis and epididymis, these were reversed by EXD treatment in a dose-dependent manner. EXD treatment was also associated with a significant increase in DJ-1 protein expression in testis and epididymis tissue samples relative to the levels observed in AZS model rats. EXD is firstly reported could significantly improve sperm motility in AZS rats and is more effective at higher dosage, even better than L-carnitine. The protective effect of EXD on sperm motility is based on the DJ-1 expression.

[Effects of sIL-13Rα2 on the nasal mucosa goblet cell apoptosis of allergic rhinitis of rats].

Objective: To investigate the effects of sIL-13Rα2 on the apoptosis of goblet cell in nasal mucosa of allergic rhinitis rats. Methods: Forty healthy male Wistar rats were randomly divided into 4 groups (10 rats per group): control group (group A), AR group (group B), sIL-13Rα2 group (group C) and triamcinolone acetonide group (group D). Ovalbumin (OVA) and aluminum hydroxide were used to establish the AR rat model. After the establishment of AR rat models, 50 µl PBS, 100 µg/50 µl IL-13Rα2 and 3.5 µg/50 µl triamcinolone acetonide were respectively dropped into each nasal cavity of every rat two times a week from 4 to 10 week in group B, group C and group D. Group A was operated with saline instead of OVA. The nasal mucosa tissues were collected at 24 h after the final administration. AB-PAS staining method was used to detect the quantity and secretion of goblet cells in the nasal mucosa tissue of all groups. Immunohistochemistry method was used to detect the expression of Bax proteins.Apoptosis was detected by TUNEL method.ANOVA analysis was used to compare multiple groups, and LSD-t test was used to compare the two groups.Pearson correlation analysis was used to analyze the correlation between the Bax positive cell rate of goblet cells and the rate of apoptotic cells. The difference was statistically significant with P<0.05. Results: Compared with group A, there were more goblet cells and hypersecretion of mucus in the nasal mucosa tissue of rats in group B while fewer in group C. The goblet cells in group C and group D were significantly fewer than that in group B (0.639 00±0.831 vs 0.956 7±0.980, 0.661 90±0.657 vs 0.956 7±0.980, t value was 2.748, 2.767, respectively, all P<0.05). The immunohistochemistry results showed that the positive expression rates of Bax protein in goblet cells of group C and group D were significantly higher than that in group B (0.880 2±0.125 vs 0.568 7±0.953, 0.938 4±0.200 vs 0.568 7±0.953, t value was -2.292, -2.685, respectively, all P<0.05). The apoptosis rates of goblet cell in nasal mucosa of group C and group D were also significantly higher than that in group B (0.516 0±0.079 vs 0.274 0±0.056, 0.535 4±0.829 vs 0.274 0±0.056, t value was -17.671, -2.225, respectively, all P<0.05). The expression of Bax protein and apoptosis of goblet cells were positively correlated (r=0.859, P<0.01). Conclusion: sIL-13Rα2 can induce apoptosis of the goblet cells in nasal mucosa of allergic rhinitis rats, by inhibiting IL-13 and up regulating Bax.

Combination of antagonistic yeasts with two food additives for control of brown rot caused by Monilinia fructicola on sweet cherry fruit.

AIMS: To evaluate beneficial effect of two food additives, ammonium molybdate (NH4-Mo) and sodium bicarbonate (NaBi), on antagonistic yeasts for control of brown rot caused by Monilinia fructicola in sweet cherry fruit under various storage conditions. The mechanisms of action by which food additives enhance the efficacy of antagonistic yeasts were also evaluated. METHODS AND RESULTS: Biocontrol activity of Pichia membranefaciens and Cryptococcus laurentii against brown rot in sweet cherry fruit was improved by addition of 5 mmol l(-1) NH4-Mo or 2% NaBi when stored in air at 20 and 0 degrees C, and in controlled atmosphere (CA) storage with 10% O2 + 10% CO2 at 0 degrees C. Population dynamics of P. membranefaciens in the wounds of fruit were inhibited by NH4-Mo at 20 degrees C after 1 day of incubation and growth of C. laurentii was inhibited by NH4-Mo at 0 degrees C in CA storage after 60 days. In contrast, NaBi did not significantly influence growth of the two yeasts in fruit wounds under various storage conditions except that the growth of P. membranefaciens was stimulated after storage for 45 days at 0 degrees C in CA storage. When used alone, the two additives showed effective control of brown rot in sweet cherry fruit and the efficacy was closely correlated with the concentrations used. The result of in vitro indicated that growth of M. fructicola was significantly inhibited by NH4-Mo and NaBi. CONCLUSION: Application of additives improved biocontrol of brown rot on sweet cherry fruit under various storage conditions. It is postulated that the enhancement of disease control is directly because of the inhibitory effects of additives on pathogen growth, and indirectly because of the relatively little influence of additives on the growth of antagonistic yeasts. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained in this study suggest that an integration of NH4-Mo or NaBi with biocontrol agents has great potential in commercial management of postharvest diseases of fruit.

Sodium Silicate Reduces Postharvest Decay on Hami Melons: Induced Resistance and Fungistatic Effects.

The effect of sodium silicate (Si) for control of decay was tested in Hami melons (Cucumis melo L. var. inodorus Jacq.). Si significantly inhibited mycelial growth of Alternaria alternata, Fusarium semitectum, and Trichothecium roseum in vitro. Si at 100 mM was more effective than Si at 25 or 50 mM at controlling the diseases caused by the three pathogens, whereas Si at 200 mM was phytotoxic. Si treatments applied at 100 mM pre-inoculation with T. roseum had lower decay incidence and severity than treatments applied post-inoculation. The protection of Si was correlated with the activation of two families of defense-related enzymes, peroxidase and chitinase. Accumulation of both enzymes was induced in fruit treated with Si and challenged by T. roseum 24 h later, and was sustained for at least 9 days in 'New Queen' and 10 days in '8601' at room temperature. It appeared that induced resistance was an important mechanism of disease control in Hami melons treated with Si.

Enhancement of biocontrol activity of yeasts by adding sodium bicarbonate or ammonium molybdate to control postharvest disease of jujube fruits.

AIMS: To assess the potential of sodium bicarbonate and ammonium molybdate as additives in enhancing the biocontrol efficacy of Rhodotorula glutinis and Cryptococcus laurentii against blue mould in jujube fruits. METHODS AND RESULTS: Two yeasts at a concentration of 107 CFU ml-1, in combination with 238 mmol l-1 sodium bicarbonate or 15 mmol l-1 ammonium molybdate, showed a significant inhibition effect on blue mould of jujube fruits stored at 20 degrees C for 5 days. The colonizing ability of the yeasts in wounded sites was significantly decreased in the presence of ammonium molybdate. CONCLUSIONS: Combining R. glutinis or C. laurentii with sodium bicarbonate or ammonium molybdate provided a more effective control of postharvest disease than using the antagonistic yeasts or the chemicals alone. SIGNIFICANCE AND IMPACT OF THE STUDY: The addition of sodium bicarbonate or ammonium molybdate reduced the number of antagonists required to efficiently control disease of postharvest fruits, which could result in the reduction of costs.

The expression of insulin-like growth factor II, hepatitis B virus X antigen and p21 in experimental hepatocarcinogenesis in tree shrews.

The purpose of this paper was to study the mechanism of synergistic effect in hepatocarcinogenesis induced by hepatitis B virus (HBV) infection and aflatoxin B1 (AFB1) intake. Immunohistochemical staining was used in formalin-fixed, paraffin-embedded sections of cancer and liver tissues. The incidence of hepatocellular carcinomas (HCCs) was 52.9% in experimental tree shrews that received both HBV and AFB1. It was significantly higher than that of animals exposed to HBV (11.1%, Group B), or (AFB1) (15.8%, group C) alone. HCC was not found in the control animals (group D). The expressions of insulin-like growth factor II (IGF-II) were 82.4%, 22.2%, 26.3% and 0 in groups A, B, C and D, respectively. The significant differences of IGF-II were observed between groups A and B, C and D (P < 0.05). The expressions of p21 were 29.4%, 11.1%, 15.8% and 0 in group A, B, C and D, respectively. The positive rate of hepatitis B x antigen (HbxAg) was significantly higher in the group A than that in the group B (52.9% vs. 11.1%, P < 0.05). The parallel relations between the incidence of HCC and the overexpressions of these genes protein have been found in each group. On the other hand, the expressions of these genes in tumour-bearing tree shrews were significantly higher than that in nontumour-bearing animals. These findings suggest a synergistic effects of HBV and AFB1 in activation of these genes in tree shrews. Overexpressions of these genes may take an important role in the course of hepatocarcinogenesis in tree shrews.

A high prevalence of p53 mutations in pre-malignant oral erythroplakia.

Oral squamous-cell carcinoma is thought to be preceded by a number of precursor stages which induce morphological changes in cells of the oral mucosa resulting in clinically detectable pre-malignant lesions such as erythroplakia or leukoplakia. To better understand the etiology of oral erythroplakia, we have examined the p53 tumor-suppressor gene (exons 5-9) for mutations in 24 oral erythroplakia lesions of varying dysplastic phenotypes by PCR/single-strand conformational polymorphism and direct DNA-sequencing analyses. A total of 12 p53 mutations were detected in 11 of 24 (46%) erythroplakia specimens (one specimen contained two different p53 mutations); 25% were single-base-pair deletions and 33% were either G:C-->T:A transversions or G:C-->A:T transitions. A high prevalence of p53 mutation was observed in all categories of erythroplakia lesions: 33% for mildly dysplastic lesions, 50% for lesions exhibiting moderate to severe dysplasia and 50% for lesions that were carcinoma in situ. Although the combined prevalence of p53 mutations observed in erythroplakia was significantly higher (p = 0.02) than that observed earlier for leukoplakia, the prevalence of p53 mutations was similar in erythroplakia and leukoplakia specimens from smokers. The prevalence and spectrum of p53 mutations observed in this series of erythroplakia lesions are similar to those observed for oral squamous-cell carcinoma. These results indicate that mutations of the p53 gene may be linked to the high malignant potential of erythroplakia and provide further evidence that p53 mutation may be an early event in the genesis of oral squamous-cell carcinoma.

Potentiation of aflatoxin B1-induced hepatocarcinogenesis in the rat by pretreatment with buthionine sulfoximine.

A single i.p. dose of aflatoxin B1 (AFB1) (1.0 and 2.0 mg/kg body wt)-induced hepatocarcinogenesis with phenobarbital as a promoter has been examined in young male Fischer rats. Immunohistochemical method has been employed to detect AFB1-induced glutathione S-transferase placental form (GST-P)-positive hepatic foci observed from 3 week and 10 week to 40-48 week periods. With 2.0 mg AFB1 dosing, the number, area and volume occupied by GST-P-positive hepatic foci increased significantly and progressively from 3 week, 10 week and 48 week periods. In long term studies (40-48 weeks), 1.0 mg and 2.0 mg AFB1 dose levels yielded linear response in area and volume occupied by AFB1-induced hepatic foci. Pretreatment of rats with L-buthionine sulfoximine (BSO), a GSH depleter, at a dose of 4 mmol/kg body wt 4 and 2 h before 1.0 or 2.0 mg AFB1 treatment enhanced the number, area and volume of GST-P-positive hepatic foci, increases being the largest at shorter time periods (3 and 10 weeks) compared to longer time periods (40 and 48 weeks). This report appears to be the first example of an enhanced chemical induced hepatocarcinogenesis in a long term study in any experimental animals species by a GSH depleting agent.

Effect of cell proliferation on initiation of aflatoxin B1-induced enzyme altered hepatic foci in rats and hamsters.

Rat is susceptible whereas hamster is resistant to aflatoxin B1 (AFB1) hepatocarcinogenesis. Effect of cell proliferation on AFB1-induced glutathione S-transferase placental form (GST-P) positive foci has been examined in these two species after a single i.p. dose of AFB1 and phenobarbital (PB) as a promoter in a 3 week period. Bromodeoxyuridine incorporation as a measure of cell proliferation and GST-P hepatic foci were analyzed by immunohistochemical methods. Hepatic cell proliferation was maximum at 24 h after either partial hepatectomy (PH) or CCl4 (4 mmol/kg) pretreatment of rats whereas cell proliferation was maximum at 48 h after PH or CCl4 (1 mmol/kg) treatment of hamsters. Enhanced number of GST-P positive hepatic minifoci (two to nine cells) and foci (>100 microns) and focal area were observed in rats with either AFB1 (0.5 mg/kg) given 24 h after PH or AFB1 (0.5 or 2.5 mg/kg) given 48 h after CCl4 dosing. In hamsters, 1 or 2 mg AFB1 treatment produced only GST-P positive single hepatocytes without presence of any minifoci whereas 3 or 6 mg AFB1 produced minifoci consisting only of doublets. Pretreatment with CCl4 48 or 72 h before 1 mg AFB1 dose level increased GST-P positive single cells and minifoci several fold. PH 24 or 48 h before 1 or 2 mg AFB1 dose level increased minifoci. However, increase in minifoci was higher in PH hamsters at 48 h compared with those at 24 h. These results indicate that even though maximum initiation occurs in both speices when AFB1 is administered at the peak of DNA synthesis, rats are more responsive than hamsters to cellular proliferation in the initiation phase of AFB1-induced hepatocarcinogenesis.

[Effects of green tea extract on the development of aflatoxin B1-induced precancerous enzyme-altered hepatocellular foci in rats].

Three kinds of extracts of green tea (decoction extract of green tea, water extract of green tea and ethanol extract of green tea) were tested for their effects on aflatoxin B1 (AFB1)-induced hepatocarcinogenesis in rats. The results revealed that all these three extracts of green tea possessed the remarkable inhibitory effects on the development of precancerous enzyme-altered hepatocellular foci. These results indicated that the main components of the green tea responsible for cancer prevention were all soluble in water and ethanol and thus providing an important clue for the search of the effective components in green tea for cancer prevention.

Reliability of a short-term test for hepatocarcinogenesis induced by aflatoxin B1.

The reliability of a short-term test for hepatocarcinogenesis induced by aflatoxin B1 (AFB1) was tested by comparing the early appearance of gamma-glutamyl transpeptidase (GGT)-positive foci with the occurrence of primary liver cancer at a later stage. All rats received a basic short-term treatment with AFB1 intraperitoneally, during which three experimental groups received Chinese green tea or 2000 or 5000 ppm butylated hydroxyanisole in the diet and a control group received basic diet. Some of the rats in each group were sacrificed at the end of the short-term procedure, and the remainder were observed up to 92 weeks. The livers of all animals were examined for GGT-positive foci or primary liver tumours. The GGT-positive foci were most numerous and largest and the incidence of liver tumours was highest in the control group. These findings suggest that GGT-positive foci are a valuable preneoplastic marker for AFB1-induced hepatocarcinogenesis, that the short-term model is fairly reliable, and that both Chinese green tea and butylated hydroxyanisole inhibit AFB1-induced hepatocarcinogenesis.

[Effect of six edible plants on the development of AFB1-induced gamma-glutamyltranspeptidase-positive hepatocyte foci in rats].

Six edible plants, green tea (GT), black tea (BT), Lentinus edodes (berk) Sing (LE), Hericium erinaceus (Bull. ex Fr.) Pers. (HE), Mixture of Ganoderma Lucidum (Ley ss ex Fr.) Karst et Ganoderma Japanium (Fr.) Lloyd (MGLJ) and mung bean (MB), were tested for the effect on the development of AFB1-induced gamma-glutamyltranspeptidase positive hepatocyte foci (gamma-GT foci) using an in vivo short-term test model in rats. The rats received intraperitoneally 12 doses of initiator AFB1, 400 micrograms/kg per dose for 2 successive weeks. Two weeks after the initiation, the rats were submitted to a modified "Solt-Farber promotion program", i.e., a two weeks' feeding of a diet containing 0.015% acetylaminofluorene plus a two-third partial hepatectomy (PH) on day 7. The rats were sacrificed 10 days after PH and the livers were processed to gamma-glutamyltranspeptidase staining. The tested substances were powdered and mixed with the basal diet at the concentration level of 30% for MB and 5% for the others. The rats were fed with the diet-containing tested substances from 10 days before the AFB1 initiation to 3 days after the AFB1 conclusion. Consequently, the liver of the rats which had consumed GT showed significantly less and smaller gamma-GT foci, and those which had consumed BT, HE and LE showed somewhat less and significantly smaller foci than the control groups. It is indicated that the four diets have an inhibiting effect on AFB1-induced gamma-GT foci in different degrees. MB and MGLJ show no significant influence on the foci.