Regional variations in the distributions of small intestinal intraepithelial lymphocytes in germ-free and specific pathogen-free mice

Previously, we reported the regional variations in intraepithelial lymphocytes (IELs) in the small intestine of mice. To clarify the effects of intestinal bacteria on the distribution of IELs, regional variations in IELs were examined using germ-free (GF) and specific pathogen-free (SPF) BALB/cA mice. The small intestine was taken and divided equally into three parts (the proximal, middle, and distal parts). IELs were isolated from each part of the intestine, and the total number of IELs in GF mice was about one seventh of that in SPF mice. The decreased number of IELs in GF mice suggests that intestinal bacteria may be essential for local expansion of IELs. On the other hand, similar regional variations in IEL subsets observed in both GF and SPF mice, except for some subsets. The similarity of regional variations in GF and SPF mice indicates that the regional variations in IEL subsets may not fundamentally depend on intestinal bacteria.     

Safety evaluation of polyphenols extracted from hop bracts.

Hop bract polyphenols contain polyphenols as promising functional ingredients. To assess the safety of topical hop bract polyphenols, Hopsphenon, we examined acute, 14-day, 28-day and 90-day toxicity tests in rats, and mutagenicity tests using Ames test and micronucleus test in mice. The acute, 14-day, 28-day and 90-day toxicity tests revealed that Hopsphenon produced no symptoms of significant injury. The lethal dose of hop bract polyphenols is greater than 2000 mg/kg. The Ames test in the absence of S9 mix for TA98 and in the presence of S9 mix for TA1537 revealed that Hopsphenon had slight mutagenicity at a high dose of 5000 microg/plate; however, in the micronucleus test, Hopsphenon was negative. These tests demonstrated that hop bract polyphenols are safe and do not cause any detrimental effects in vivo under the conditions investigated in this study.     

HER2/neu expression in bladder cancer: relationship to cell cycle kinetics

BACKGROUND AND OBJECTIVES: The incidence of overexpression of HER2/neu in bladder cancer is one of the highest among all human malignancies tested; such overexpression is thought to play a role in the aberrant proliferation of cancer cells. This study was conducted to evaluate the quantitative assessment of HER2/neu expression by enzyme immunoassay (EIA) and its prognostic significance in differentiating between high and low proliferating tumors. PATIENTS AND METHODS: Tissue samples were collected from 35 patients with benign bladder lesions, 28 with bilharzial bladder cancer, and 25 with nonbilharzial bladder cancer. Twenty normal samples were obtained from normal safety margin areas in nonbilharzial bladder cancer patients. Out of the malignant samples, 22 were found to be squamous cell carcinoma and 31 were transitional cell carcinoma. All samples were examined for HER2/neu expression by EIA and Western blot (WB). Flow cytometric (FCM) analysis was also performed in all the samples provided. RESULTS: HER2/neu was found to be significantly overexpressed in the malignant group compared to the benign and normal groups (P < 0.001) and no significant difference was found between the bilharzial and nonbilharzial cancer groups or between the transitional and squamous cell carcinoma groups. HER2/neu was significantly correlated to ploidy (P = 0.001), synthetic phase fraction, SPF (P = 0.012), and DNA index (P = 0.002). No significant correlation was found between HER2/neu and stage or grade while it was significantly associated with lymph node status of the tumour (P = 0.02). CONCLUSION: HER2/neu can be measured reliably by the EIA method as confirmed by WB. The quantitative assessment of HER2/neu expression in malignant tumors aided by other proliferation markers such as SPF, DI, and ploidy could be useful in selecting patients for more aggressive treatment or for predicting outcome.     

Deletions of p15 and p16 in schistosomal bladder cancer correlate with transforming growth factor-alpha expression

OBJECTIVES: Cell proliferation is stimulated by growth factors and inhibited by p15 and p16 gene products. We compared cell regulators, TGF-alpha, p15, and p16, in schistosomal and non-schistosomal bladder cancer to explore possible differences in their alterations between the two subtypes and their correlations with proliferation pattern [synthetic phase fraction (SPF)], DNA ploidy, and clinicopathological factors. METHODS: Tumor tissue samples were obtained from 120 patients. Expressions of p15 and p16 genes were investigated by the polymerase chain reaction, while TGF-alpha protein expression was measured by an enzyme immunoassay (EIA) method. RESULTS: Deletion of both p15 and p16 was observed in 62 and 46 bladder tumors, respectively. TGF-alpha was overexpressed in 64 bladder tumors. A highly significant association was observed between the two deleted genes and TGF-alpha positivity. Of the entire group, p15 and p16 alteration and positive TGF-alpha (> or =cutoff value) were significantly expressed in schistosomal bladder cancer (68.1%, 60.9%, and 65.2%), and squamous cell carcinoma type (SCC) (69.1%, 64.7% and 72.1%) compared to those with non-schistosomal bladder cancer (29.4%, 7.8%, and 37.3%) or transitional cell carcinoma (TCC) (28.8%, 3.8%, and 28.8), respectively. A significant association between p15 and p16 deletion and TGF-alpha positivity with high SPF, aneuploid DNA pattern, late stages, and high histological grades was also documented. CONCLUSION: Alteration of p15 and p16 genes and overexpression of TGF-alpha appears to be an event in bladder cancer that occurs more frequently in schistosomal bladder cancer and SCC, and may play an important role in their development. These observations may provide insight into treatment guided by molecular changes.     

防晒化妆品抗水性能的仪器测定法

[目的]研究采用SPF-290分析仪及一种新型载体聚甲基丙烯酸甲酯板对防晒产品的抗水性能进行评价的可行性。[方法]对15个经人体试验证实具有抗水性能的防晒化妆品用SPF-290分析仪及振荡水槽进行测定。[结果]抗水性的测试结果与相应的人体试验结果相当吻合,符合率为14/15。[结论]用本法取代人体法来测定防晒化妆品的抗水能力是可行的。     

SPF小型猪血液学、血液生化正常参考值、尿常规值测定

本实验选用20头60dSPF小型猪,雌雄各半,对其26项血液常规、生化正常值和18项尿常规值进行测定,并将雌雄SPF小型猪血液常规、生化常值、尿常规值进行比较.结果表明SPF小型猪血、尿值变动幅度更小,SPF小型猪血液成分变化趋势与文献报道SPF大、小鼠血液成分变化趋势一致,红细胞(RBC)总数、血红蛋白(Hgb)量较高,白细胞(WBC)总数、中性粒细胞(NEUTRO)较低.淋巴细胞(LYM)、中性粒细胞分叶型(NEUTROl)、血清球蛋白(GLOb)、白球比(ABG)相差非常显著(P<0.01);淋巴细胞、白球比雌性高于雄性,中性粒细胞分叶型、血清球蛋白雄性高于雌性;血小板总数(PLT)、酸性粒细胞(EO)、血液pH值(pH)、血清白蛋白(ALB)、血清氯(Cl)、血清钠(Na)、血清钙(Ca)、血清磷(P)相差显著(p<0.05);血液pH值、血清白蛋白、血清钙、血清磷雌性高于雄性;血小板总数、酸性粒细胞、血清氯、血清钠雄性高于雌性;其他血液常规、生化常值及18项尿常规值,雌雄SPF小型猪相差不显著(p>0.05).     

SPF鸡不同生长阶段粪便菌群组成及多样性研究

目的 提供SPF鸡不同生长阶段肠道菌群的组成及多样性数据.方法 采集10日龄、12周龄、23周龄和52周龄SPF鸡粪便,利用Illumina HiSeq 2500高通量测序方法,对样品16S rRNA的两个高变区(V3-V4)进-测序分析,并进行了α-多样性指数、β-多样性指数和粪便群落特征分析.结果 四个生长阶段粪便样品菌群具有相似的较高的丰富度和良好的均匀度,物种多样性无显著差异(P＞0.05);厚壁菌门(Firmicutes)、变形菌门(Proteobacte ria)和拟杆菌门(Bacte roidetes丰度占95％以上;乳杆菌属(Lactobacillus)、链球菌属(Streptococcus)、拟杆菌属(Bacteroides)、肠球菌属(Enterococcus)为优势菌属,不同时期丰度差异达显著水平(P＜0.05).结论 四个生长阶段菌群组成多样,且优势菌群主要为有益菌.该实验结果能够为SPF鸡品质评定、健康监测和安全性评价提供依据.     

Acute and subchronic toxicity studies of seabuckthorn (Hippophae rhamnoides L.) oil in rodents

Seabuckthorn (Hippophae rhamnoides L.) has been traditionally used as medicine and nutritional supplement for a long period of time. However, information on the systemic toxicity and safety evaluation of seabuckthorn and its extracts is still scarce. The purpose of this study was to evaluate the potential toxicity of seabuckthorn oil by an acute oral toxicity study in mice and a 90-day repeated oral toxicity study in rats. No mortality or signs of toxicity was observed in mice treated with 20 mL/kg body weight seabuckthorn oil in the acute toxicity study. In the subchronic toxicity study, 80 Sprague-Dawley rats (10 animals per sex per treatment group) were administrated with 10, 5, 2.5 and 0 (control) mL/kg body weight of seabuckthorn oil daily for 90 days by gavage. There were no signs of toxicity and treatment-related changes in rats treated with seabuckthorn oil on mortality, body and organ weights, food consumption, blood biochemistry and hematology, gross necropsy and histopathological examinations. Based on the finding of this study, the maximum tolerated dose of seabuckthorn oil was >20 mL/kg for mice for acute toxicity study, and the no-observed-adverse-effect level was 10 mL/kg body weight for both male and female rats for 90-day toxicity study.     

Acute toxicity, 28-day repeated-dose toxicity and toxicokinetic study of timosaponin BII in rats

Timosaponin BII (TBII), a major steroidal saponin isolated from Anemarrhena asphodeloides Bge., displays a variety of promising pharmacological activities, such as neuroprotection, enhancement of learning and memory, vascular protection and inhibition of platelet aggregation; therefore, it has been developed as a pharmaceutical for prevention or treatment of dementia. Given the safety concerns surrounding timosaponins and the absence of studies on the safety of TBII, the potential toxicity of TBII was evaluated in toxicity and toxicokinetic studies in rats. In the acute oral toxicity study, loose stools were observed in rats receiving 4000 mg/kg, and the symptoms recovered within 1 day. In the 28-day repeated-dose oral toxicity and toxicokinetic study, rats receiving 540 mg/kg showed loose stools and a slight deceleration of body weight growth in both sexes, and the females also showed a slight decrease in food consumption. Moreover, urinalysis indicated reversible treatment-related toxicity in rats receiving 540 mg/kg. The toxicokinetic study demonstrated a dose-dependent increase in systematic exposure to TBII after 28 successive days of oral treatment with TBII. The accumulation coefficients of TBII were 4.35, 1.70 and 1.81, respectively, in rats that received 60, 180 and 540 mg/kg. The no-observed-adverse-effect level (NOAEL) is proposed to be 180 mg/kg.     

Characterization of the immune response to the major glycoprotein (gp71) of friend leukemia virus III. Influence on endogenous MuLV-mediated pathogenesis

The response of AKR mice to immunization with purified gp71 of Friend murine leukemia virus (FLV)⁹ was characterized. A humoral response was detectable by radioimmune precipitation assay with FLV or radioimmunoassay with FLV gp71. In contrast, no reactivity with either the endogenous AKR virus or AKR gp71 was detectable. The humoral immune response to FLV gp71 was also detectable in complement-dependent cytotoxicity assays and again appeared type-specific for FLV-producing cell lines and could be specifically blocked by absorption with FLV gp71. In contrast, no specific neutralization of FLV was detectable with immune sera. Cell-mediated reactivity was examined by blastogenesis assays with FLV gp71 and in vitro cytotoxicity assays. Although no consistent cell-mediated cytotoxicity was demonstrable in vitro, speen cells from FLV gp71-immunized mice did undergo blastogensis when incubated with purified gp71. The ability of FLV gp71 immunization to protect AKR mice from spontaneous thymomas and irradiated C57B1/6 mice from radiation-induced thymomas was also examined. The results demonstrate that FLV gp71 immunization has no clear influence on induction of radiation thymomas in C57Bl/6 mice. In contrast, FLV gp71 immunization greatly increased the rate of mortality of AKR mice from thymic lymphomas without altering the time of onset of disease-related deaths. The possible mechanisms behind this enhancement of thymomas by FLV gp71 immunization, including the activation of the endogenous AKR virus, are considered.