Abstract:Objective To investigate the feasibility of high fat-introduced hyperlipeidemia model in male SD rat and study the time rule of molding. Methods 30 Male adult rats of SD Strain bred in the animal house of the institute were divided into 3 groups after 1 week adaptation, group 1:control group, normal diet; group 2:model 1 group, high fat high cholesterol diet;group 3:model 2 group, high fat high cholesterol diet. The period of experiment was 8 weeks. Food and water intake were measured everyday and body weight were measured every four days. Blood were collected by orbital venous at the end of fourth,sixth,eighth week to test their serum lipid level.At the end of experiment,animals were killed to collect liver and aorta tissue for HE stain. Results Compared with control group,the food intake of model 1 was higher and model 2 was significant lower,water intake of model 2 was significant lower,the ratio of liver/weight of two model groups were significant heavier,and weight of model groups were higher.High fat diet significantly increased TC levels of model groups at the end of fourth,sixth week. The level of LDL-c in model 1 group were higher and the HDL-c were lower compared with control group.HE stain showed the livers of control group were regular,arrangements of the liver cells were trim, dyeing present uniformity. The two model groups showed a large range of hepatocyte fatty change,a few liver blood sinus were in congestion and infiltrated with inflammatory cells. Aorta HE stain showed no significant change among 3 groups. Conclusions The method of high fat-introduced hyperlipeidemia model in male SD rat is feasible and the model turned out to present hypercholesterolemia with severe fatty liver.On the other hand,levers of serum lipid increased within an increase-inter-adjustment-increase state.In the process of modeling,how to overcome the symptom of anorexia and the state of cholesterol inter-adjustment in animals is the key to successfully establish hyperlipeidemia model.

Abstract:Objective To investigate the effect of Smad3 on cell migration of A549 and HeLa cells. Methods Primers for pCMV-Myc-Smad3 plasmid construction and siRNA targeting Smad3 were designed and synthesized. pCMV-Myc-Smad3 plasmid was constructed with molecular cloning techniques. Overexpression of Smad3 with Myc-tag or silencing of endogenous Smad3, and then scratch assay was used to detect the migration ability of A549 and HeLa cells in vitro. Results pCMV-Myc-Smad3 plasmid was successfully constructed. Overexpression of Smad3 significantly up-regulated the migration rate of A549 and HeLa cells. Conversely, in the same cells, silencing of endogenous Smad3 or treatment with Smad3 inhibitor, SIS3, down-regulated the migration rate. Conclusions Smad3 promotes cell migration of A549 and HeLa cells.

Abstract:Objective Erlotinib (Trade Name:Tarceva) is a new targeted drug for the treatment of non-small cell lung cancer (NSCLC) that has a wide clinical application in recent years, but commonly carries many side effects, among which the most common and unbearable one is rash. The aim of this study is to observe the changes of epidermis, pathology, immunohistochemistry and other aspects before and after the application of Tarceva in mice and try to copy the rash animal models caused by Tarceva and thus to provide models for the clinical topical medications of rash. Methods 20BALB/c female mice were randomly divided into four groups.The experimental group(Ⅱ、Ⅲ、Ⅳ group) was given 100 mg/kg dosage with the concentration of 10 g/L erlotinib solution by gavage, and the control group (I group)with an equal volume of deionized water by gavage once daily. The hairs from the head,neck and back of each mouse were removed 24 hours prior to the administration,and at the end of the experiment, clipping the skin in the neck, back and waist,then observed differences between the experimental group and the control group in mice skin, biopsy, immunohistochemistry and the like. Results (1) There were statistically significance (P<0.01 or P<0.05) in the four groups of mice in five aspects as hair regrowth days, days of the complete regrowth of hair, desquamation time, the time of appearance of rash and the number of pore expansion;(2) Ki67:There were no statistically significant differences among the four groups(P>0.05).Conclusions (1) This experiment confirms many researchers' point of view that "the rash induced by EGFRIs is an inflammatory response"(2) A mouse model of rash induced by FGFRIs is successfully established,and this is a reliable,practical and reproducible model which applies to a large number of establishment of "EGFRIs drug-induced rash in animal models",and can be popularized for clinical,experimental and institutional uses.

Abstract:Objective To compare the biological characteristics of several different anxiety rat models established by different methods of stress at different time points and provide experimental basis for the most appropriate modeling methods. Methods 60 rats were randomly divided into normal, empty bottle stress, chronic emotional stress (CES) group, restraint stress for 3h, 6h, and modeling respectively. In the experimental 7 d, 14 d, 21 d, elevated plus maze and fear condition system was used to test anxiety-like behavior in rats, open field test to study anxiety or depression-like behavior, forced swimming test was used to detect depression-like behavior in rats, and using the Elisa test kit to detect the contents of 5-HT, DA in the hippocampus in rats. Results Anxiety-like behavioral test results showed that rats in empty bottle stress, CES, 6 h restraint stress group started to have anxiety-like behavior since 14 d, then anxiety-like behavior was becoming increasingly apparent. Forced swimming test results showed that immobility time in 6 h restraint rats was significantly increased in the first 7 d(P<0.05). Meanwhile, compared with control group, hippocampal 5-HT, DA contents in empty bottle stress and CES rats increased significantly since 14 d. Conclusions Among several stress methods established anxiety model, anxiety-like behavior in 3 h restraint stress was not obvious; 6 h restraint stress exhibited a depression-like behavior in the forced swimming test might be due to prolonged stress. Empty bottle stress and CES can successfully establish the anxiety rat model, and the anxiety behavior of the rats have some differences. Corresponding model methods can be selected according to different experimental purposes.

Abstract:Objective Treatment and histopathological observation of demodex canis in beagle dogs. Methods Using the method of direct smear for microscopic examination of demodex canis. Histopathological observation on the skin of the parasitic parts after routine paraffin section and HE staining. Results (1) Clinical observation:The red spots and hair removal was appeared on limbs, eyes, lower abdomen and other parts of the skin of canine patients.The skin of the limbs becomes thicker and wrinkles. (2) Blood routine examination:Basically normal. (3) Microscope observation:The results showed that a large number of worms and eggs of small demodex canis could be found. (4) Histopathological observation:Hair follicles showed a large number of demodex mites and eggs. The sebaceous glands and sweat glands have normal morphology and no mites was found. A large number of eosinophils and neutrophil infiltration were seen around the hair follicles. It was also found that the formation of multifocal granuloma:the granuloma was oval shaped. (5) Treatment programme:The combination of medication and the strengthening of environmental control has been shown to be effective. Conclusions Granuloma caused by demodex canis can be divided into immune granuloma.It may not be possible to destroy the sebaceous glands after infection with small demodex canis.Whether the sebaceous gland is infected with the demodex canis may be associated with the worm species or course of disease.

Abstract:Objective To study the effect on ovarian tumor migration invasion after inhibition the expression of IGFBP2, IGFBP3.Methods siRAN interference IGFBP2, IGFBP3 expression in ovarian cancer cell lines SKOV3, SKOV3 proliferation detected by CCK-8 kits,SKOV3 apoptosis detected by flow cytometry,SKOV3 migration and invasion detected by transwell experiment and scratched cell healing detection; CCK-8 method detected survival after treated different concentrations of cisplatin (5, 10, 15, 20 ug/mL). Results The proliferation ability of SKOV3 dropped and apoptosis increased after treated siRAN IGFBP2, IGFBP3 compared with the control group, migration and invasion function decline, resistance level to improve greatly. Conclusions The expression of IGFBP2, IGFBP3 in ovarian cancer affected migration and invasion.

Abstract:Objective To establish a simple and reliable experimental rodent model sensitive to coxsackievirus A16 (CVA16).Methods Mongolian gerbils with different age were selected and inoculated intraperitoneally with live CVA16, and the gerbils were observed daily until 14 days postinoculation to screen for the most optimal ages sensitive to the virus. The dose-dependent symptoms were evaluated and the 50% lethal dose (LD50) was determined. The virus titers were measured in blood and various tissues of CVA16-infected Mongolian gerbils 3 days post-infecton. Finally, the gerbils were immunized twice with inactivated CVA16 vaccine at day 1 and day 11, respectively, followed by challenge with the virus with a dose of LD50 at day 14. The gerbils were then observed for another 2 weeks to record their body weight, symptom and mortality rate. Their blood samples were collected from the eyes, and CVA16-specific neutralizing antibody titers and total antibody titers was checked by microneutralization test and ELISA, respectively. Results Various clinical symptoms, such as inactivity, hind limb weakness, paralysis and even death occurred in gerbils following CAV16 infection. 7-day-old and 14-day-old gerbils are susceptible to CVA16 infection whereas 28-day-old gerbils are resistant. The most sensitive and appropriate age is 14-day-old. The 50% lethal dose was determined to be 1×104.5 CCID50. High titers of the virus were confirmed in blood and various tissues of Mongolian gerbils contracted CAV16 3 days post-infecton. The survival rate is 87.5% for 14-day-old gerbils preimmunized with two doses of inactivated CVA16 vaccine and challenged with the virus. The geometric mean titers (GMTs) of neutralizing antibody was 28.14, and the seroprevalence was 87.5%. Conclusions Mongolian gerbils is sensitive to CVA16 and the virus reproduces actively in Vivo. Thus, it can be used as a reliable small animal model for studies of CVA16 pathogenesis, vaccine development and drug evaluation.

Abstract:Objective To investigate the natural infection rate of Mouse Adenovirus(Mad)in Guangdong province and to estimate the tissue distribution and serology changes of Mad in infected mice. Methods Serums of mice from general environment of Guangzhou in 2007 and 2015, and the SPF mice from12 supervision and 32 entrusted units of Guangdong province among 2013~2015 were detected by ELISA.Thirty-six 3 weeks old BALB/c mice were each intraperitoneally inoculated with 0.2 mL of Mad in 4.5×10⁶ copies/μL concentration. The tissue samples, including heart, liver, spleen, lung, kidney, brain, stomach, cecum contents and serum were taken at 0 d before inoculation, and 3, 7, 10, 15, 18, 21, 30, 37, 44, 51, 60 d after inoculation (3 animals each time).Real-time fluorescent quantitative PCR (Q-PCR) method was used to detect the virus nucleic acid of tissue. In addition, the antibody against Mad was detected by ELISA.Results Antibody positive rate was 24.44%~84.15% and 0% among the general environment mice and the SPF mice, respectively, which is given priority to with Mad-2 K87 serotype strains. There were no clinical manifestations among all infected mice.The virus nucleic acid positive rate was 100% (3/3) in all tissue samples in 3 d and 7 d, which maintained a longest time to 60 d in spleen. Besides the liver, virus nucleic acid content were highest at 7 d in each tissue of infected mice, with spleen highest (5.5×10⁵ copies/μL), heart was second at 3.4×10⁵ copies/μL, cecum contents and stomach ranked third (2.6×10⁵ copies/μL), followed by brain (0.8×10⁵ copies/μL). The antibody could be successfully detected at 15 d after inoculated, hit the peak at 37 d and maintained a high level until 60 d.Conclusions Mad infection rate is low in SPF mice, high in general environment mice.Mad nucleic acid content is the highest and positive rate maintain the longest in spleen,suggesting that Mad mainly replicate in spleen. And 7 d is the best pot for nucleic acid testing.Serum neutralizing antibody can be used as a monitoring index within 15~60 d after infection.

Abstract:Objective Establish premature ovarian failure (POF) model in female Sprague Dawley by tripterygium wilfordii, and investigate the effect of bushenyangxue prescription on the levels of anti-mullerian hormone (AMH). Methods After POF model was established, rats were gave by gavage of different dosage of Bushenyangxue prescription for 30 days. The changes of histomorphology on rat ovarian tissue were observed by hematoxylin-eosin staining. Serum AMH concentraction, protein and mRNA expression of AMH were measure with ELISA, immunohistochemical staining and qRT-PCR, respectively. Results The follicle and corpus luteum were atrophied after tripterygium wilfordii challenge, which was improved after treatment with Bushenyangxue prescription. Serum AMH, protein and mRNA expression of AMH were decreased tripterygium wilfordii-treated rats; this decrease was inhibited after treatment with Bushenyangxue prescription. Conclusions Our study indicates that Bushenyangxue prescription could preserve the AMH levels of POF rats. These findings suggest that Bushenyangxue prescription may be a useful strategy to treat POF.

Abstract:Objective In this study we investigated the analgesic effect of oxysophocarpine (OSC) on carrageenan-induced inflammatory pain in mice and to explore its possible mechanism. Methods Mouse ear swelling test and carrageenan-induced paw edema were used to investigate the effects of OSC (10, 40 and 80 mg/kg) on inflammatory pain in mice. The mechanical hyperalgesia and allodynia were measured by von Frey filaments. Paw edema was assessed by toe volumetric measuring instrument. text the content of TNF-α, IL-1β, IL-6 and PGE₂.Results OSC showed a significant anti-inflammatory effect in the mouse ear swelling test. OSC (10, 40 and 80 mg/kg) also significantly reduced the paw edema volume and improved mechanical allodynia threshold value on carrageenan-induced inflammatory pain, as well as relieved paw tissues inflammatory damage and reduced the numbers of neutrophils in mice. OSC significantly suppressed over-expression of TNF-α, IL-1β, IL-6 and PGE₂. Conclusions Based on these findings we propose that OSC attenuates inflammatory pain by suppressing the Pro-inflammatory cytokines.

Abstract:Objective To analyze the literature characteristics on environmental enrichment published in China, and provide reference for researchers.Methods The papers relevant to environmental enrichment published in CNKI database and Wanfang database were statistically analyzed from the aspects of publishing year, journal of publication, author, institution and subjects covered by the article with bibliometrics method. Results There were 422 papers published before Dec 31,2015, in which 273 papers published in 126 kinds of journals. The number of papers increased gradually, especially after 2006. The top 3 journals with the most articles published were Chinese Journal of Wildlife(28),Chinese Journal of Rehabilitation Medicine(15) and Chinese Journal of Behavioral Medicine and Brain Science(12). Xinqiao Hospital of TMMU and Fujian Medical University Union Hospital accounted for the top 2 total number of the papers, from which Yan-hui Chen and Cong-min Zhao published the more articles. Those papers covered 17 subjects, neurology, pathergasiology, biology and pediatrics were the main subjects. There were 3 kinds of animal that got enriched, the first was laboratory animals(69.6%), then zoo animals(22.9%),and farm animals(7.5%). Conclusions The research of environmental enrichment was relatively concentrated in terms of neurology and psychiatry. To improve animal welfare was mainly in the zoo animals; laboratory animal research in this direction was less.It is necessary to strengthen the work in this regard.

Abstract:Objective To evaluate the sensitizations of Hydroxyethyl Starch 40 Sodium Chloride Injection. Methods Active systemic anaphylaxis(ASA) test, internal sensitive index's determination and screening and Passive Cutaneous Anaphylaxis(PCA) test are carried out. Observe the allergy safety of the samples. Results We use guinea pigs to carry on ASA test and PCA test with Hydroxyethyl Starch 40 Sodium Chloride Injection from 45 batches of 3 companies and no changes have occurred. Measuring and comparing the sensitive index of HIS,IgG, IgM and IgE in plasma, IgM and IgE are not obvious variation; HIS and IgG positive group and negative group and sample groups are different.Conclusions It is suggestion that Hydroxyethyl Starch 40 Sodium Chloride Injection safety evaluation should include allergic reactions, HIS and IgG sensitive index monitoring in clinical application of hydroxyethyl starch.

Abstract:Objective To explore the regulatory effect of TAK1 inhibitors on MAPK and NF-κB signaling pathway in diabetic rats and its renal protection mechanism. Methods A total of 48 rats accorded to the random number table method were divided into DN group, TAK1 group and control group,each group with 16 rats,control group with normal fed,DN group and TAK1 group by the intraperitoneal injection of 1% 50 mg/kg STZ DN model rat.8 rats were killed in each group at 4 weeks and 8 weeks respectively,the pathological changes of renal tissue were observed,serum TNF-α,MCP-1,IL-1β levels were detected by enzyme linked immunosorbent assay,p38MAPK,NF-κBp63 protein expression were detected by Western blotting,p38MAPK、NF-κBp63 mRNA levels in renal tissue were detected by real time fluorescence quantitative PCR. Results At 4 weeks and 8 weeks, the body weight, blood glucose and UAER of DN group and TAKl group were significantly higher than those in control group (P<0.05). The body weight and UAER of DN group were significantly higher than those of TAK1 group (P<0.05).The serum TNF-α,MCP-1,IL-1β levels in DN group and TAK1 group were significantly higher than those in control group(P<0.05),and DN group serum TNF-α,MCP-1,IL-1β levels were significantly higher than those in TAK1 group (P<0.05).The expression levels of p38MAPK,NF-κBp63 protein and mRNA in DN group, TAK1 group were significantly higher than those in control group (P<0.05), and p38MAPK,NF-κBp63 protein and mRNA in DN group was significantly higher than that in TAK1 group (P<0.05).Conclusions TAK1 induces inflammation by activating MAPK and NF-κB signaling pathways,and participates in diabetic renal injury.TAK1 inhibitors with anti-inflammatory effect by down regulate the expression of inflammatory factors.

Abstract:Objective To investigate the effect of Icariin (ICA) experimental IgA nephropathy in rats and to explore related mechanisms. Methods Experimental IgA nephropathy rat model was established and then model rat were treated with or without different doses of ICA. Then, urine RBC, Urine protein and urine NAG were analyzed; IgA precipitation was detected with immunofluorescence staining; the protein level of NF-κBp65 and MCP-1 were examined by immunohistochemical staining; the mRNA level of IL-4, IL-10 and IL-13 were determined by quantitative PCR. Results The concentrations of urine RBC, Urine protein and urine NAG were reduced after ICA treatment, as companied by a decrease of IgA precipitation. Moreover, ICA treatment also decreased the protein level of NF-κBp65 and MCP-1, and the mRNA level of IL-4, IL-10 and IL-13. Conclusions ICA exerts a certain degree of efficacy on the treatment of experimental IgA nephropathy through regulating NF-κBp65 and MCP-1 expression and the immunoregulation mechanism.

Abstract:Objective To improve the method of paraffin section and frozen section of adipose tissues. Methods The adipose tissues were collected and quickly placed in 10% neutral formaldehyde. Sections were prepared by setting different dehydration procedures. The slices were observed under microscope after Hematoxylin-Eosin staining. The expression of PPARγ and ITLN1 in adipose tissues was detected by immunohistochemistry. For frozen sectioning, the adipose tissues were collected and quickly placed in 10% neutral formaldehyde or Calcium formaldehyde, After embedded by OCT, put in -80℃ refrigerator for at least 30 minutes. Setting the temperature of frozen section cabinet to -20℃ and the temperature of sample to -30℃. Results Following the improved methods, the slices of adipose tissues were better than before. The structural integrity and consistency of adipose tissues were preserved well, the staining was distinct, and the improved methods had no effect on immunohistochemistry results. The improvement of the frozen sectioning had greatly improved the slice quality, the adipose tissues showed complete structure and good morphology. Conclusions The improved methods can be successfully applied to different experimental animals, which provide a powerful condition for the study of adipose tissues.

Abstract:Objective We established a rapid detection method of Pasteurella spp. and provided a reference for microbiological quality control of laboratory animal. Methods According to the β subunit of bacterial RNA polymerase (rpoB) protein multiple alignments of 13 different Pasteurella spp. published in NCBI. The degenerate primers were designed by CODEHOP designer online. CODEHOP PCR method was applied to detecting Pasteurella spp. after the specificity and sensitivity of the method had been evaluated by 21 reference strains. Results Standard strain amplified fragment were about 200 bp by degenerate primers PastF6/PastR5. The primers are able to distinguish between Pasteurella spp. and the other pathognic organisms of laboratory animal respiratory tracts. Sensitivity of this method were 0.2 pg/μL~2 pg/μL to different Pasteurella. The Pasteurella positive rate was 19.1% in 609 animal's respiratory samples. The accuracy of positive results was 100% through verifying by sequenced and blast. Conclusions The established method has good specificity and sensitivity. It can be used to detect Pasteurella spp. in animal samples.

Abstract:Cancer is a group of heterogeneous disease caused by diverse genomic alterations in oncogenes and tumor suppressor genes. Despite recent advances in high-throughput sequencing technologies and development of targeted therapies, novel cancer drug development is limited due to the high attrition rate from clinical studies. Patient-derived xenografts (PDX) models are generated by implanting sectioned patient tumor fragments into immunodeficient mice. PDX models retain many of the key characteristics of patients' tumors including histology, genomic signature, cellular heterogeneity, and drug responsiveness. These models cannot only serve as a platform for co-clinical trials by enabling the integration of clinical data, genomic profiles, and drug responsiveness data to determine precisely targeted therapies, but also be applied to the development of biomarkers for drug responsiveness and personalized drug selection. This review summarizes our current knowledge of this field, including methodologic aspects, applications in drug development, challenges and limitations, and utilization for precision cancer medicine.