[Protective effect of Wuzi Yanzong recipe on testicular DNA damage and apoptosis in natural ageing rats].

To study the protective effect of Wuzi Yanzong recipe on testicular DNA damage and apoptosis in natural ageing rats, SPF grade 16-month-old SD male rats were randomly divided into three groups: ageing model group, low and high dose Wuzi Yanzong recipe groups (WZ, 1， 4 g·kg⁻¹). In addition, 2-month-old SD rats were used as adult control group (10 rats in each group). The ageing model group and the adult control group were fed with normal diet for 4 months. Wuzi Yanzong groups received medicated feed for 4 months. After fasting for 12 hours, the rats were sacrificed. Then testis tissues were taken and weighed to calculate the testis index. The change of testicular tissue morphology was observed by HE staining. Expression and localization of DNA damage-associated protein ATR were observed by immunofluorescence. The expressions of DNA damage-related proteins γ-H2AX, Chk1, p-p53 and apoptosis-related proteins Bcl-2 and Bax in testes were detected by Western blot. The apoptosis of testis tissue in rats was detected by using TUNEL. The results showed that as compared with the youth control group, the protein expression levels of γ-H2AX, Chk1, p-p53 and Bax were significantly increased while Bcl-2 protein expression level was significantly decreased intestis tissues of ageing model group. Wuzi Yanzong recipe significantly decreased protein expression levels of γ-H2AX, Chk1, p-p53 and Bax and increased Bcl-2 protein expression level as well as Bcl-2/Bax ratio. Immunofluorescence results showed that Wuzi Yanzong recipe could significantly decrease the ageing-induced ATR, increase in testis tissues. TUNEL results showed that Wuzi Yanzong recipe could significantly attenuate the germ cell apoptosis in testicular tissues. All the above results suggest that Wuzi Yanzong recipe could protect the germ cell in testicular tissues of natural ageing rates from DNA damage and apoptosis, and the mechanism may be associated with regulating p53 signaling pathway.

Oleanolic acid rejuvenates testicular function through attenuating germ cell DNA damage and apoptosis via deactivation of NF-κB, p53 and p38 signalling pathways.

OBJECTIVES: Inflammation can cause degenerative changes of reproductive function. Oleanolic acid (OA), the effective component from Ligustrum lucidum Ait., exhibits significantly anti-inflammation and antiageing activity. However, whether OA restores testicular dysfunction via inhibition of inflammation with ageing is unclear. Here, in a natural ageing rat model, we investigated the protection effects of OA and its mechanism of action. METHODS: Eighteen-month-old Sprague Dawley (SD) rats were randomly divided into ageing control group and two OA-treated groups (5 and 25 mg/kg). Nine-month-old SD rats were used as adult controls. All rats were received either vehicle or OA for 6 months. Then, histomorphology, weight and index of testis, protein expression and immunohistochemistry were examined. KEY FINDINGS: Oleanolic acid significantly restored testicular morphology and improved testicular weight and index. Moreover, OA significantly inhibited phospho-NF-κB p65 and its downstream proinflammatory cytokines' expressions, including IL-1ß, COX-2 and TNF-α in testis tissues. Similarly, OA remarkably inhibited IL-1ß and TNF-α production. OA significantly attenuated germ cells' DNA damage and apoptosis. Such changes were accompanied by downregulation of γH2AX, p-P53 and Bax expressions, and upregulation of Bcl-2 and Bcl-2/Bax ratio. In addition, OA remarkably inhibited p38 signalling. CONCLUSIONS: Oleanolic acid effectively rejuvenates testicular function via attenuating germ cell DNA damage and apoptosis through deactivation of NF-κB, p53 and p38 signalling pathways.

[Application of Best Subsets Regression on the risk classification for Spermophilus Dauricus Focus].

OBJECTIVE: To study the risk classification of animal plague in Spermophilus Dauricus Focus, using the Best Subsets Regression (BSR) model. METHODS: Matlab, BSR and exponential smoothing were employed to develop and evaluate a model for risk classification as well as to forecast plague epidemics at the Spermophilus Dauricus Focus. Data was based upon the Inner Mongolia surveillance programs. This model involved 7 risk factors, including density of Spermophilus dauricus, percentage of hosts infested, host flea index, percentage of nests infested, nest flea index, percentage of runways infested, and runway flea index. RESULTS: Forecasting values of the classification model(CM)were calculated and grouped into 3 risk levels. Values that over 2/3 of the CM would indicate the existence of potential epidemics while those below 1/3 would indicate that there were no risk for epidemics but when values that were in between would indicate that there exist for high risk. Annually, during the observation period in the Inner Mongolia Spermophilus Dauricus Foci, the detection of Yersinia pestis gave a risk rating value of 1 which stood for existing epidemics, while nil detection rate generated a 'zero' value which representing the situation of non-epidemic. The overall plague epidemics forecasting surveillance programs in 2012 at the Spermophilus Dauricus Foci indicated that no active plague was observed. When the forecasting values became over 2/3, combinations of all the risk factors would achieve the consistency rates of 100%. When the forecasting values were below 1/3, combinations of at least the first 4 factors could also achieve the consistency rates of 100%. However, when the forecasting values fell in between, combinations of at least the first 4 factors would achieve the consistency rates of around 50%. CONCLUSION: Results from our study showed that plague would not be active to become epidemic, in 2012.

Analysis of methylene blue and its metabolites in blood by capillary electrophoresis/electrospray ionization mass spectrometry.

A method for the determination of methylene blue (MB) and its metabolites (azure A, azure B and azure C) in rat blood by CE-electrospray ionization mass spectrometry (CE-ESI-MS) was developed in this paper. Different analytical parameters were investigated in detail such as pH and concentration of separation buffer, and ESI-MS instrumental parameters. Under the optimum conditions, MB and its metabolites were separated and detected in 27.3 min. LODs (defined as S/N=3) of this method were 0.22, 0.25, 0.10 and 0.30 µg/mL for MB, azure A, azure B and azure C, respectively. To get a satisfactory extraction efficiency of MB and its metabolites in rat blood, different extraction solutions were studied. By using this method, MB and its metabolites (azure A, azure B and azure C) were successfully analyzed in rat blood samples.