ObjectiveTo explore the mechanism through which Gantou Fumu decoction （GDFMT）-containing serum inhibits the activation of hepatic stellate cells. MethodsSerum pharmacology techniques were employed to prepare GDFMT-containing serum samples. Rat hepatic stellate cells （HSCT6） were randomly allocated into three groups： Control， penicillamine （90 mg·kg^-1）， and GDFMT （6.8 g·kg^-1）. Cell viability was assessed by the cell counting kti-8 （CCK-8） assay， intracellular ROS levels were determined by flow cytometry using the DCFH‑DA fluorescent probe and while extracellular collagen Ⅰ （ColⅠ） levels were quantified via enzyme linked immunosorbent assay （ELISA）. Intracellular α-smooth muscle actin （α-SMA） expression was visualized through immunofluorescence staining. Real-time polymerase chain reaction（Real-time PCR） and Western blot were performed to evaluate alterations in the expression of nuclear factor-erythroid 2-related factor 2 （Nrf2）， glutamate cysteine ligase， catalytic （GCLC）， glutamate-cysteine ligase， modifier subunit （GCLM）， Kelch-like ECH-associated protein 1 （Keap1）， NAD（P）H：Quinone oxidoreductase 1 （NQO1）， and heme oxygenase-1 （HO-1）. Genetic knockdown of Nrf2 was achieved through small interfering （siRNA） transfection to validate pathway specificity. ResultsCCK-8 data indicated that under the selected experimental conditions， culturing HSCT6 cells with 10% （volume fraction） GDFMT-containing serum for 48 h demonstrated equivalent effect in inhibiting HSCT6 cell proliferation compared with culturing HSCT6 cells with 10% （volume fraction） penicillamine-containing serum for 36 h， with no statistically significant difference between the two groups. After treatment of HSCT6 cells with siRNA-mediated Nrf2 gene transfection technique， the inhibitory effect of GDFMT on the activation process of HSCT6 cells was significantly weakened. After HSCT6 cells were cultured with GDFMT-containing serum for 48 h， the flow cytometry with a fluorescence probe showed that the content of reactive oxygen species （ROS） in the cells was significantly reduced. After the expression level of the Nrf2 gene in HSCT6 cells was knocked down by siRNA transfection， the content of ROS increased （P<0.01）， indicating that GDFMT significantly reduced the ROS level in activated HSCT6 cells， and this effect strictly depended on the integrity of the Nrf2 pathway. Quantitative analysis by ELISA showed that the synthesis of ColⅠ in the HSCT6 cell culture system treated with GDFMT-containing serum for 48 h was lower than that in the control group （P<0.01）. After the Nrf2 gene level of HSCT6 cells was knocked down by siRNA transfection， the ColⅠ concentration in the culture supernatant increased （P<0.01）. The results of α-SMA detection by IF method showed that compared with the normal group， GDFMT-containing serum reduced the expression of α-SMA （P<0.01） and the protein level of α-SMA in HSCT6 cells was increased （P<0.01） after siRNA-mediated Nrf2 silencing， suggesting that Nrf2 signaling pathway may be involved in regulating the phenotypic transformation process of the cells. Real-time PCR and Western blot results showed that GDFMT-containing serum reduced the protein level of α-SMA （P<0.01） and enhanced the gene and protein levels of Nrf2 in the nucleus and GCLC， HO-1， Keap1， GCLM， and NQO1 in the cytoplasm of HSCT6 cells （P<0.05， P<0.01）. After the Nrf2 gene level of HSCT6 cells was knocked down by siRNA transfection， the protein level of α-SMA was increased （P<0.01）， and the levels of GCLC， GCLM， Keap1， NQO1， and HO-1 were decreased （P<0.05， P<0.01）. These results indicated that GDFMT-containing serum successfully triggered the Nrf2 signaling pathway of HSCT6 cells. ConclusionThe anti-hepatic fibrosis effect of 10%GDFMT was equivalent to that of penicillamine. In addition， GDFMT can inhibit HSC activation and reduce oxidative stress damage by activating the Nrf2/antioxidant response element （ARE） pathway， as manifested by the up-regulated expression of Nrf2 and downstream antioxidant genes （GCLC， GCLM， HO-1， Keap1， and NQO1） and reduced secretion of ROS， α-SMA， and ColⅠ.

Objective:To evaluate the effects of fine particle matter(PM2.5)and ozone(O3)com-bined exposure on adenosine triphosphate(ATP)amount and ATPase activities in nasal mucosa of Spra-gue Dawley(SD)rats.Methods:Twenty male SD rats were divided into control group(n=10)and exposure group(n=10)by random number table method.The rats were fed in the conventional clean environment and the air pollutant exposure system established by our team,respectively,and exposed for 208 d.During the exposure period,the concentrations of PM2 5 and O3 in the exposure system were moni-tored,and a comprehensive assessment of PM2 5 and O3 in the exposure system was conducted by combi-ning self-measurement and site data.On the 208 d of exposure,the core,liver,spleen,kidney,testis and other major organs and nasal mucosal tissues of the rats were harvested.Each organ was weighed and the organ coefficient calculated.The total amount of ATP was measured by bioluminescence,and the ac-tivities of Na+-K+-ATPase and Ca2+-ATPase were detected by spectrophotometry.The t test of two inde-pendent samples was used to compare the differences among the indicator groups.Results:From the 3rd week to the end of exposure duration,the body weight of the rats in the exposure group was higher than that in the control group(P＜0.05),and there was no significant difference in organ coefficients be-tween the two groups.The average daily PM2 5 concentration in the exposure group was(30.68±19.23)μg/m3,and the maximum 8 h ozone concentration(O3-8 h)was(82.45±35.81)μg/m3.The chemi-luminescence value(792.4±274.1)IU/L of ATP in nasal mucosa of the rats in the exposure group was lower than that in the control group(1 126.8±218.1)IU/L.The Na+-K+-ATPase activity(1.53±0.85)U/mg in nasal mucosa of the rats in the exposure group was lower than that in the control group(4.31±1.60)U/mg(P＜0.05).The protein content of nasal mucosa in the control group and the exposure group were(302.14±52.51)mg/L and(234.58±53.49)mg/L,respectively,and the ac-tivity of Ca2+-ATPase was(0.81±0.27)U/mg and(0.99±0.73)U/mg,respectively.There was no significant difference between the groups.Conclusion:The ability of power capacity decreased in the rat nasal mucossa under the sub-chronic low-concentration exposure of PM2 5 and O3.

Copper plays a very important role in the physiological activities of the body. Copper metabolism disorders due to various reasons can disrupt the copper homeostasis of the body to cause a deficiency or excess of copper,resulting in a series of pathological processes that eventually lead to cell death and organ dysfunction. Cuproptosis,a newly discovered cell death mechanism,is related to copper overload. Cuproptosis has been confirmed in Wilson's disease,which is of great significance for future research on the mechanism and treatment of copper metabolism disorders.

Objective:To evaluate the effects of exposure of fine particle matter (PM 2.5) and ozone (O 3) in Beijing as the main pollutants on olfaction of SD rats. Methods:In October 16, 2018, twenty 8-week-old SD rats were randomly divided into two groups, 10 rats in the exposure group and 10 rats in the control group. They were fed in air pollutant exposure system and clean experimental environment respectively, and the concentrations of PM 2.5 and O 3 in each system were measured. The degree of olfaction damage of SD rats at different feeding time was assessed by using the buried food test (BFT). The difference of BFT time between the two groups was analyzed by performing the repeated measures analysis of variance. Results:The results showed that the concentrations of PM 2.5 and O 3 in the exposure group were (22.65±11.47) μg/m 3 and (12.36±5.87) μg/m 3, respectively, while those in the control group were both 0 μg/m 3. The repeated measures analysis of variance showed that the time of BFT in the exposure group was longer than that in the control group ( F=6.49, P=0.031). With the increase of feeding time, the time of BFT was prolonged ( F=61.69, P<0.001). Conclusion:Exposure to PM 2.5 and O 3in the atmosphere might lead to olfaction damage in rats.

Objective:To evaluate the effects of exposure of fine particle matter (PM 2.5) and ozone (O 3) in Beijing as the main pollutants on olfaction of SD rats. Methods:In October 16, 2018, twenty 8-week-old SD rats were randomly divided into two groups, 10 rats in the exposure group and 10 rats in the control group. They were fed in air pollutant exposure system and clean experimental environment respectively, and the concentrations of PM 2.5 and O 3 in each system were measured. The degree of olfaction damage of SD rats at different feeding time was assessed by using the buried food test (BFT). The difference of BFT time between the two groups was analyzed by performing the repeated measures analysis of variance. Results:The results showed that the concentrations of PM 2.5 and O 3 in the exposure group were (22.65±11.47) μg/m 3 and (12.36±5.87) μg/m 3, respectively, while those in the control group were both 0 μg/m 3. The repeated measures analysis of variance showed that the time of BFT in the exposure group was longer than that in the control group ( F=6.49, P=0.031). With the increase of feeding time, the time of BFT was prolonged ( F=61.69, P<0.001). Conclusion:Exposure to PM 2.5 and O 3in the atmosphere might lead to olfaction damage in rats.

Objective Application of Case-based teaching method (CTM) in the class of Experimental Animal Quality Control aims to break the traditional teacher-centered and textbookcentered teaching model,to invoke students' interest and creativity by integrating practice and theory,and to cultivate their capacity in identifying,analyzing and solving problems.Methods According to different requirements of teaching contents,students were organized to discuss practical cases about quality control of experimental animals through problem-centered learning style.Results Students' capacity was enhanced in terms of problem solving with acquired knowledge,and the ability to pursue new knowledge,the intriguing and vivid cases stimulated students' interest,activated classroom atmosphere,and helped improve teaching efficiency.Conclusion Experimental animal quality control is a complicated system; Case-based teaching method leads to higher efficacy of the course of Experimental Animal Quality Control.

In recent years,many kinds of animal models of human diseases with different genetic background were developed.Some were established in China,others were imported from abroad.Precious were these animal models,their biological features could be lost for various reasons,for instance,reverse from mutation,lost of modified or constructed gene,etc.This study provided some suggestions for scientific management of the animal models to in terms of quarantine inspection,specific feeding method and methods to maintain gene stability,so as to preserve their biological features.