Abstract:Objective In China, the chromium-containing Chinese traditional medicine, Tianmai Xiaoke tablet (TM), is used in the treatment of type 2 diabetes mellitus. However, its mechanism of action is not fully clear yet. The aim of this study was to investigate the effect of TM on glucose metabolism in diabetic rats and to identify whether TM has a direct effect on islet cells through microRNAs. Methods Thirty-two healthy 5-month old SPF male Sprague-Dawley rats were divided into control group, diabetic group, low dose TM group (TML), and high dose TM group (TMH). The miRNA expression profiles of pancreas samples were analyzed using microRNA array and verified by Q-PCR. Results Eight-week treatment with TM significantly decreased the fasting blood glucose in the diabetic rats. The blood glucose was significantly reduced in the TM-treated groups before and after oral glucose administration. Fasting insulin and HOMA-IR were suppressed in the TM-treated groups. miR-448, let-7b, miR-540, miR-296, miR-880, miR-200a, miR-500, miR-10b, miR-336, miR-30d, miR-208, let-7e, miR-142-5p, miR-874, miR-375, miR-879, miR-501, and miR-188 were up-regulated, while miR-301b, miR-134, and miR-652 were down-regulated in the TMH group. Through target gene analysis and real-time PCR verification, we found that these miRNAs, especially miR-375 and miR-30d, could stimulate insulin secretion of the pancreatic islets. Conclusions Our findings suggest that TM can effectively decrease FBG, enhance the sensitivity to insulin, and also modulates lipid metabolism in diabetic rats. TianMai Xiaoke Tablet may improve the blood glucose and alleviate the insulin resistance in diabetic rats through up-regulation of pancreatic miR-375 and miR-30d, promote the proliferation of islet β-cells and inhibit the proliferation of α-cells, increase the insulin gene expression, up-regulate the pancreatic let-7b, let-7e, miR-142-5p and miR-375, and suppress the function of cytokine-receptor interaction pathway and MAPK pathway.

Abstract:Objective To establish a rat model of immunological hepatic injury and to explore the effect of blocking or activating Tim-3 pathway on Th17 cells in the rats. Methods Forty SPF male Wistar rats (body weight 180±20 g) were used in this study. The rat model of immunological hepatic injury was established by injecting 12.5 mg/kg concanavalin A (Con A) through the tail vein once a week for eight weeks. Splenic lymphocytes were isolated. Serum was separated and stored in -20℃. Liver tissue samples were fixed in 4% paraformaldehyde for further study. The splenic lymphocytes were divided into 5 groups: blocking group, blocking control group, activating group, activating control group and ConA control group. The Tim-3 pathway was blocked by anti-Tim-3 while activated by recombinant galectin-9, and cultured 72 h under 37℃, 5% CO₂ environment. The cell culture supernatants were saved at -20℃ for later use. Biochemical analysis was performed to determine the expression of serum ALT, AST and ALB. Morphological changes of the liver were examined by histopathology using HE staining. Immunohistochemical staining was used to detect the expression of IL-17A and ROR-γt proteins in the liver tissues. Enzyme-linked immunosorbent assay (ELISA) was used to assess the IL-17A and IL-6 in the cell supernatants while real time PCR was applied to determine the expression of ROR-γt mRNA in splenic lymphocytes. Results Compared with the control group, inflammatory cell infiltration, hepatic tissue injury and many pseudolobules were easily found in the liver tissue, and immunohistochemical examination showed significantly increased levels of IL-17A and ROR-γt proteins in the liver tissues of the model group. The expressions of IL-17A and IL-6 in the blocking group were increased than that of the blocking control group (P <0.05), while the levels of IL-17A and IL-6 in the activating group were lower when compared with that of the activating control group (P<0.05). Compared with the blocking control group, the real time PCR showed that the expression of ROR-γt mRNA in the blocking group was significantly increased (P<0.05). Conclusions The pathogenesis of immunological hepatic injury is closely related to Th17 cells. Immune-regulating Tim-3 pathways may affect the effect of Th17 cells then further participate in the mechanism of immunological hepatic injury.

Abstract:Objective To compared two different methods, the mechanical allodynia test and cold hyperalgesia test in a rat model of oxaliplatin-induced peripheral neuropathy. Methods Oxaliplatin frequently causes severe acute and chronic peripheral neuropathies. In this experimental study, 40 healthy female Wistar rats (body weigh 210±10 g) were used. We used Von Frey filament test to determine the 50% paw withdrawal threshold and mechanical allodynia in rats, and also used acetone spray and cold plate to examine the pain behavior in rats. Results Compared with the control group, the rats of model group showed a 50% decrease of the paw withdrawal threshold (P<0.05), appearance of allodynia and hyperalgesia (P<0.05), and increase of the number of paw withdrawals caused by cold stimulation (P<0.05), but there were no obvious differences in the response to heat stimulation in rats of both groups. Those results were similar in general to the clinical manifestations in patients.Conclusions Both the two ways of mechanical allodynia test are good choices. von Frey filaments with bending forces of 4 g and 15 g are a better way to simulate the test of mechanical allodynia and hyperalgesia in clinical patients, while the up-down method can objectively and quantitatively reflect the reduction of paw withdrawal threshold. The acetone spray test allows us to immediately record the reaction of rats after cold stimulation. Our findings may be useful for further studies of oxaliplatin-induced neuropathy.

Abstract:Objectives To establish a simple, inexpensive and efficient technique for in vitro culture of monocyte-derived macrophages (MDM) from rhesus macaques of Chinese origin. Methods Peripheral blood of healthy rhesus macaques (Macaca mulatta) were obtained in heparinized vacutainer collection tubes. Peripheral blood mononuclear cells (PBMCs) were isolated from blood by Ficoll gradient centrifugation. Serum was isolated from peripheral blood of the autologous animals. PBMCs were plated in 48-well-plate (3×10⁶ cells/well) or 96-well-plate (0.8×10⁶ cells/well) for 24 h. After removal of non-adherent cells from the culture, monocytes were cultured in RPMI 1640 supplemented with different proportions (2%, 4%, 8%, 10%) of autologous serum or fetal calf serum (FCS) for 7 days. To examine the biological function of MDM, lipopolysaccharide (LPS) was added to MDM culture to determine inflammatory cytokine production. Also, MDM cultures were tested for the susceptibility to simian immunodeficiency virus (SIV) or simian-human immunodeficiency virus (SHIV) infections. Results The cell cultures with RPMI1640 containing 2% autologous serum yielded the best results with regard to macrophage morphology, the response to LPS stimulation and susceptibility to SIV or SHIV infection. The purity of adherent macrophages under condition of 2% autologous serum culture was higher than 96%. Conclusions RPMI 1640 with 2% autologous serum is suitable for culture in vitro of peripheral blood monocytes from rhesus macaques. This technique is simple, inexpensive, no need for growth factor and highly effective to obtain adherent and well differentiated macaque monocytes. Therefore, this method provides an important tool for culture of macaque AIDS viruses and for related immunological research.

Abstract:Objective To explore the oxidative stress in Parkinson's disease dyskinesia with Yin deficiency stirring wind pattern, and the interventional effect of compound formula rehmannia on the disease condition. Methods One hundred and twenty healthy male SD rats (body weight 180-200 g) were randomly divided into four groups: the control group, sham-operated group, model group, and Rehm group. The rat model with Parkinson's disease (PD) was established by injection of 6-hydroxyl dopamine to destroy the substantia nigra using the brain stereotaxic apparatus. Dyskinesia was induced in the PD rats by injection of levodopa into the brain. The dyskinesia PD model rats received intraperitoneal injection of levodopamine 50 mg/kg and benserazide 12.5 mg/kg for 2 weeks to induce Yin deficiency stirring wind pattern. The rats of Rehm group were given gastric gavage of compound formula rehmannia 2 mL/kg once a day for 4 weeks or 6 weeks. At the end of experiment (4 weeks and 6 weeks), neurobehavioral examination was performed, and the contents of SOD, MDA, GSH, and GSH-Px in the striatum tissue were assessed by colorimetry. Results In the LID group, the contents of SOD, GSH, GSH-Px in the striatum tissue were significantly lower than those in the normal control group and sham-operated group, and the malondialdehyde (MDA) content showed an increasing trend. In the compound rehmannia prescription group, the concentrations of SOD, GSH, GSH-Px in the striatum tissue showed an increasing trend, even more obvious along with the time course, and the malondialdehyde (MDA) content showed an decreasing trend, also even more obvious along with the time course. Conclusions The compound rehmannia prescription can improve the oxidative stress in rat models of Parkinson's disease dyskinesia with Yin deficiency stirring wind pattern. The interventional effect of compound rehmannia prescription may be mediated by scavenging free radicals and reducing the damage to the cells, thus, to alleviate the symptoms of Parkinson's disease dyskinesia.

Abstract:Objective To create an optimal method for establishment of a rat model of constipation-predominant irritable bowel syndrome (IBS-C), and evaluate the traditional Chinese medicine (TCM) syndrome in this generated IBS-C model. Methods One hundred and thirty-three SPF neonatal rats of 20 litters were used in this study. Using maternal separation (MS), ice water stimulation, water limitation, MS combined with ice water stimulation, and MS combined with water limitation to establish rat constipation model. The Chinese medicine Xiao-Yao powder was administered to the rat models. To observe the general behavior, body weight and daily food-intake, to measure the defecation numbers and water content of the feces within 4 hours, and the types of feces were assessed by Bristol stool scale. Colorectal distention test was used to assess the intestinal sensitivity, and histological examination using haemotoxylin staining was performed to observe the alterations of intestinal tissues. Results MS combined with ice water stimulation was the best method to induce the rat models with decreased defecation number and decreased water content of feces. There were no obvious histological alterations in the intestinal tissue. The scores by Bristol stool scale indicated that the model was an IBS-C model with visceral hypersensitivity, and the results of Xiaoyao powder administration indicated that this IBS-C model met the requirement of clinical characteristics of liver-stagnation and spleen-deficiency syndrome. Conclusions Maternal separation combined with ice water stimulation is a good method to successfully establish an IBS-C rat model with liver-stagnation and spleen-deficiency, highly simulating the clinical symptoms of this syndrome.

Abstract:Objective To establish a BALB/c mouse model of influenza B virus infection, and to provide a foundation for studying the pathogenesis, mechanism of transmission of influenza B virus infection and developing new vaccines against influenza B virus. Methods Using genetic synthesis and reverse genetic technology, influenza B virus was rescued in vitro. We used the whole genome sequencing approach to validate the identity between the rescued viral genome sequences and the sequences reported in Genbank. To establish the BALB/c mouse model of influenza B virus infection, BALB/c mice were infected with 10⁵ EID₅₀ dose of the rescued virus, and the weight change, survival rate, and viral replication in the lungs were analyzed. Results We successfully rescued influenza B virus B/Yamagata/16/88 in vitro, and this virus was named B-S9. The genome sequencing results showed that the genome sequences of B-S9 was consistent with the GenBank-reported sequences. BALB/c mice were artificially infected with B-S9, and no death due to infection was observed. The above results indicated that B-S9 is of low pathogenicity to the BALB/c mice. The mice infected with B-S9 showed body weight decline in 3 days post inoculation (dpi) but restored in 8 dpi. The virus titers could be detected in the lungs of mice infected with B-S9 on dpi 3 and dpi 6, respectively. Furthermore, the virus titer in the mouse lungs on dpi 3 was 132 times higher than that on dpi 6. Conclusions A reverse genetic system of influenza B virus B/Yamagata/16/88 is successfully established, and a BALB/c mouse model of influenza B virus infection is established. To date, studies of influenza B virus are limited at home and abroad. The establishment of this reverse genetic system provides not only a platform for studying the pathogenesis and mechanism of transmission of influenza B virus, but also provides a way for developing new live-attenuated influenza B virus vaccine.

Abstract:Objective To investigate the changes of adenosine triphosphatase activity, expression of calmodulin (CaM), and calmodulin-dependent protein kinase II (CaMK II) protein in CaMK II signaling pathways in the liver tissue of rats with spleen-qi deficiency. Methods 3-month-old SPF Wistar rats (male:female=1:1) were randomly divided into 4 groups: normal control group, and three spleen-qi deficient model groups (observed on days 7, 14 and 21), 12 rats in each group. The spleen-qi deficient rat models were generated by bitter-cold purgatives, exhaustion and fully or poorly feeding food in turn, and their general conditions and ATPase activity were evaluated. Confocal laser scanning microscopy was used to detect cellular [Ca²⁺]i concentrations and Western blotting was used to test CaM, CaMK II, p-CaMK II expression in liver tissue of the spleen-qi deficient rats. Results Compared with the normal group, general condition was poor, activities of Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase were significantly decreased (P<0.01, P<0.05), and expressions of CaM, CaMK II, and p-CaMK II in the liver tissue were decreased. A most obvious difference was observed in the spleen-qi deficient model day 21 group (P<0.01). Conclusions Our results demonstrate that decease of adenosine triphosphatase activities and [Ca²⁺]i concentration is involved in the spleen-qi deficiency, and expressions of key molecules of CaMK II signaling pathway, CaM, CaMK II and p-CaMK II proteins, are also decreased.

Abstract:Objective To compare and analyze the sequence between Tibet minipigs and Pubmed pig (NM_214256.1) IGF-1 cDNA by cloning and sequencing the IGF-1 gene cDNA of the liver tissue of Tibet minipigs. Methods Total RNA was extracted from Tibet minipig liver and amplified the sequence of IGF-1 gene by RT-PCR. The amplified fragment was cloned into pMD18-T vector, and the recombinant plasmid pMD18-T-IGF-1 was constructed and sequenced. Results Tibet minipig IGF-1 gene fragment containing the coding region of 567 bp. The coding region encoded 186 amino acids, and 99% were homologous compared with that of the pig (NM_214256.1). Compared with pig (NM_214256.1), we found that the mutation in the 440 bp, 455 pb, occurred at G→A, C→T mutation, and the corresponding amino acids were changed, histidine→arginine and leucine→serine, respectively. Conclusions The results of this study provide a molecular basis for breeding research. Whether the mutation at the two sites changes the protein function and body size or not need to be further studied.

Abstract:Objective To develop a rapid, specific and sensitive fluorescence real-time RT-PCR assay for detection of murine norovirus (MNV). Methods A pair of primers and a Taqman probe were designed targeting highly conserved sequences among MNV strains,which are located at the open reading frames 1 (ORF1)-ORF2 junction region. The internal control(IC)was also designed and constructed to determine false-negative results. A real-time RT-PCR assay with the IC was established for MNV detection by optimizing reaction components and conditions. The standard curve was plotted based on the linear relationship between the amount of plasmid DNA and cycle threshold(Ct)values. In addition,the specificity, sensitivity and reproducibility of the assay were evaluated. 344 clinical samples were used to determine the efficacy of this real-time RT-PCR method. Results The specificity test showed that this real-time RT-PCR assay could specifically detect MNV and had no cross reactions with mouse hepatitis virus, Theiler's murine encephalomyelitis virus, Sendai virus, pneumonia virus of mice, reovirus III, Hantaan virus and lymphocytic choriomeningitis virus. The standard curve showed fine linear relationship between the amount of plasmid DNA and Ct values (correlation coefficient r²=0.9986). The developed assay was sensitive enough to detect as low as 10 copies/μL. The lower quantification limit of MNV was estimated as 1.78×10^-2 TCID₅₀/mL,which was 10 times more sensitive than conventional RT-PCR method, and 100 times more sensitive than virus isolation method. Both the intra-batch and inter-batch coefficients of variation were less than 2% based on 5 repeated intra-batch and inter-batch test of 5 samples. 344 clinical samples were tested by this developed real-time RT-PCR assay and showed a positive ratio of 29.94% (103/344). Conclusions This real-time RT-PCR assay is a specific, sensitive and reproducible assay. Moreover,the internal control in the real-time RT-PCR system can be used to determine false negative results. This assay could be exploited for routine detection, clinical diagnosis and epidemiological survey of mouse norovirus.

Abstract:Objective To isolate microsatellite DNA markers from genome of Mugilogobius chulae for further identification of its genetic background. Methods A microsatellite-enriched genomic library for Mugilogobius chulae was constructed by magnetic-bead enrichment method using biotinylated (AC)₁₅ probe. After a series of steps, the obtained microsatellite markers were assessed and evaluated versus that of wild population of Mugilogobius chulae. Results 74 repeated sequences containing microsatellite sequences were obtained from 95 randomly picked positive clones. 33 primers were designed and synthesized. All of these loci were selected for polymorphism analysis in a wild population of Mugilogobius chulae, and 12 of them showed polymorphism. The mean number of alleles (Na), the number of effective alleles (Ne), observed heterozygosity (Ho), expected heterozygosity (He), and polymorphic information content (PIC) were 2.9167, 2.2311, 0.4583, 0.5633 and 0.4474, respectively. Conclusions The microsatellites developed in this study represent the first nuclear markers described in Mugilogobius chulae and will be useful tools for genetic studies of Mugilogobius chulae.

Abstract:Objective To observe the effect of high sucrose/high fat diet and streptozotocin (STZ) on glucose and lipid metabolism, liver histology and protein kinase B (PKB) phasphorylation in Chinese Banna miniature pigs, and to investigate its molecular mechanism.Methods Eight healthy 2-month old male Banna minipigs were randomly divided into control and diabetes groups, 4 pigs in each group. Diabetic minipigs were induced by high sucrose/high fat diet and STZ and the levels of fasting blood glucose, insulin, total cholesterol, triglycerides were measured per month for 12 months. At the end of 12^th month, the minipigs were sacrificed and liver histology was examined using HE staining and the hepatic lipid accumulation was observed using Sudan IV staining. Real-time PCR and Western blot were used to determine the expression of PKB mRNA and protein, and the phosphorylation level of PKB-Ser⁴⁷³ in the liver tissue. Results At the end of 12th month, the minipigs of model group showed hyperglycemia, insulin deficiency and dyslipidemia compared with that of the control group (P<0.05).The fat degeneration was increased, and glycogen was decreased in the liver tissue of model group. High sucrose/high fat diet and STZ increased the PKB mRNA and protein expression, while decreased PKB phosphorylation in the liver tissue (P<0.05). Conclusions High sucrose/high fat diet and STZ can induce insulin resistance (IR) and diabetes mellitus, promote lipid and cholesterol accumulation, and inhibite hepatic glycogenogenesis, which may be related with inhibition of PKB phosphorylation.

Abstract:Objective To investigate the effect of the Chinese medicine Jinlida on triglyceride-related genes in skeletal muscle of fat-induced insulin resistant ApoE^-/- mice. Methods Eight male C57BL/6J mice were set to the normal group (group A). Forty male ApoE^-/- mice were fed high-fat diet for 16 weeks and divided into model group (group B), rosiglitazone (group C), Jinlida low dose group (group D), Jinlida moderate dose group (group E), Jinlida high-dose group (group F), and giving gavage for 8 weeks. TG content in the skeletal muscle was determined by enzymatic and BCA protein concentration assay. Oral glucose tolerance test (OGTT) was used to evaluate the degree of insulin resistance in the mice. Real-time fluorescent quantitative reverse transcription PCR (RT-PCR) and Western blot method were used to determine the expressions of hormone-sensitive lipase (HSL), adipose triglyceride lipase (ATGL), and peroxisome proliferator-activated receptor gamma (PPARγ) mRNA and proteins in the skeletal muscle. Results Jinlida to varying degrees lowered the fasting blood glucose (FBG), cholesterol (TC), triglyceride (TG) and low density lipoprotein cholesterol (LDL-C), increased the high density lipoprotein (HDL-C), reduced fasting insulin (FIns) level and raised insulin sensitive index (ISI), and significantly improved the abnormal glucose tolerance in the mice. Jinlida to a certain degree raised the levels of HSL, ATGL, PPARγ mRNA and protein expressions. Conclusions Jinlida can alleviate fat-induced insulin resistance in ApoE^-/-mice through regulation of triglyceride-related gene expression.

Abstract:Objective To explore the expression and correlation of fractalkine and TLR4 in atherosclerotic plaques in the common carotid artery of apoE^-/- mice. Methods Twenty-four ApoE^-/- mice were divided into three groups, fed with standard chow diet (SD), high-fat diet (HF) or HF and atorvastatin (HF+A).12 weeks later, the blood lipids were checked. The plaque area and vascular stenosis rate of the common carotid artery were measured. All the data were used to evaluate the severity of atherosclerotic lesions of the animals. Moreover, immunohistochemical staining was used to examine the levels of fractalkine and TLR4 expressions. Results The plaque area and vascular stenosis rate of the common carotid artery in the HF group were larger than those in the SD group (about 5-fold and 3-fold), but only vascular stenosis rate of the HF+A group was significantly reduced, compared with the HF group (35.27±3.84 vs. 27.02±2.69, P<0.05). The expressions of fractalkine and TLR4 were significantly increased in the HF group (3.24±0.96 vs. 10.69±2.11, 1.29±0.57 vs. 9.32±1.02), and decreased in the HF+A group (10.69±2.11 vs. 5.73±1.30, 9.32±1.02 vs. 3.32±0.51)(P<0.05). Conclusions The expressions of fractalkine and TLR4 are changed synchronously in apoE^-/- mouse atherosclerotic plaques. These results suggest that fractalkine and TLR4 may have some inherent relationship, which may play an important role in the development of atherosclerosis.

Abstract:Objective To explore the tetracycline-inducible expression system in zebrafish application strategy and build a transgenic zebrafish with tetracycline-inducible and liver-specific expression of green fluorescent protein. It is a fundamental work to construct conditional gene function studies and tissue-specific transgenic zebrafish disease models. Methods In this experiment pfabp10-rtTA was constructed with rtTA under the liver-specific promoter fabp10. pTRE-Tight-BI-AcGFP1 and pfabp10-rtTA were co-transfected into HeLa cells to confirm the doxycycline induction, and after that two plasmids were injected into zebrafish 1-cell stage embryos. After 30 μg/mL doxycycline incubation, the integrated individuals stably expressing GFP in the liver were selected. Results HeLa cells were transfected with pfabp10-rtTA and pTRE-Tight-BI-AcGFP1 plasmids and cultured with 1 μg/mL doxycycline, and take 0 μg/mL concentration of doxycycline as control. Western blot results showed that GFP expression was significantly higher in the experimental group than in the control group. The selected stable integrated zebrafish were cultured at a concentration of 30 μg/mL doxycycline conditions, with obvious GFP expression in the liver, while in the control group without doxycycline conditions, no obvious GFP expression was seen in the liver. Conclusions Our findings indicate that the use of the tetracycline-inducible expression system in liver of transgenic zebrafish lines could provide a good animal model tool for establishing conditional transgenic zebrafish disease models, and serve the research such as liver organogenesis and development of liver diseases.

Abstract:Objective To observe the effect of circadian rhythm on local cerebral ischemia-reperfusion injury in rats. Methods One hundred SPF male SD rats (body weight 320±20 g) were raised in light(6:00-18:00)-dark(18:00-6:00)cycle environment for 4 weeks. Then all rats were randomly divided into 6:00 group, 12:00 group, 18:00 group and 24:00 group. Rat models of cerebral ischemia-reperfusion injury were induced by middle cerebral artery occlusion at corresponding time points for each group. The neurological symptoms were evaluated, cerebral infarction area and SOD and MDA levels in brain tissue were determined, intracellar Ca²⁺ concentration of brain tissue was measured, the level of serum endothelin (ET) and VEGF were assayed, and the pathological changes of brain tissue at 24 h after ischemia-reperfusion were examined. Results All rats showed typical neurological symptoms, which were milder in the 24:00 group than that of the 6:00 group, and the cerebral infarction areas of the 6:00 group and 12:00 group were significantly larger than that of the 18:00 group and 24:00 group. The SOD activities of 18:00 group and 24:00 group were significantly higher than that of the 6:00 group and 12:00 group, and the MDA levels of 18:00 group and 24:00 group were significantly lower than that of the 6:00 group and 12:00 group. The concentration of intracellular Ca²⁺ of the 24:00 group was lower than that of 6:00 group and 12:00 group, and the concentration of intracellular Ca²⁺ of the 18:00 group was significantly lower than that of the 6:00 group. The contents of serum ET in the 18:00 group and 24:00 group were significantly lower than that in the 6:00 group. The content of serum VEGF of 24:00 group was obviously lower than that of the 6:00 group and 12:00 group. The brain pathological changes of the 18:00 group and 24:00 group were obviously milder than that of the 6:00 group and 12:00 group. Conclusion Circadian rhythm may significantly influence the cerebral ischemia-reperfusion injury in rats.

Abstract:Objective To evaluate the therapeutic effect of a Chinese medicine, Shangke Jiegu Tablet, on the rat model of bone fracture. Methods After anesthesia, complete transverse fracture of the middle portion of the radius in the right forelimb was made in rats. The rats were randomly divided into three groups: model group, treatment group and sham operation group as control. Each group included 20 rats, half male and female. The rats in the treatment group were intragastrically administered the drug in a dose of 0.33 g/kg once daily for 4 weeks, while the model and sham operation groups were given normal saline. After the end of drug administration, images of X-ray, CT and MRI were taken to record the healing of the bone fracture. Finally the rats were sacrificed by anesthesia, and the bones were taken for anti-rupture strength test and routine histopathological examination. Results The imaging examinations clearly showed dense callus formation at the fracture site, and the fracture line was blur or disappeared. Calcium deposition was observed in most fracture sites. It was consistent with the results of histopathological observation. The anti-rupture strength of the healed bones was also obviously increased in the treatment group. Conclusions Medical imaging techniques can be used to more objectively evaluate the therapeutic effects of drug therapy for bone fracture. Especially, the 4-dimensional CT imaging can more clearly and directly reveal the healing process of bone fracture, and is worth of popularization and application.

Abstract:Objective To investigate the changes of hypothalamic-pituitary-adrenal (HPA) axis secretory function in male and female castrated rats, and to confirm that sex hormones can affect the function of HPA axis and explore if this effect has relationship with gender and the lasting of time.Method Eighty SPF SD rats (half male and female) aged 8 weeks, body weight 180-220 g, were randomly divided into the male model group, male control group, female model group and female control group, 20 rats in each group. After one week adaptation feeding, rats in the male and female model groups were castrated. At 3 and 13 weeks after castration, blood samples were collected from the heart apex, and serum content of corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and cortisol (CORT) were assayed by using enzyme-linked immunosorbent assay (ELISA) and statistically analyzed. Results At 3 weeks after castration, the serum contents of CRH and CORT in the male model group were significantly lower than those in the male control group (P<0.01), and serum ACTH level in the male model group was significantly lower than that in the male control group (P<0.05). In the female model group, the serum contents of CRH and CORT were lower than those in the female control group (P<0.05), and the level of serum ACTH showed a decreasing tendency. At 13 weeks after castration, serum CORT level in the male model group was significantly lower than that in the male control group (P<0.01), while other hormones showed no obvious changes. The serum CORT content in the female model group was lower than that in the female control group (P<0.05), while other hormones changed not obviously. At 13 weeks after modeling, compared with that at 3 weeks after castration, serum CRH, ACTH and CORT contents had no significant differences compared with those in the male and female model groups (P>0.05). Conclusions Castration causes dysfunction of the HPA axis, and this influence exits in both male and female rats. The effect of androgen on male rats is greater than that of estrogen on female rats. Castration can cause a drop of peripheral blood CRH, ACTH and CORT levels. Presumably, androgen may be more conducive to secretion of CRH, ACTH and CORT than that on females. In the condition of low sex hormone levels, the secretion of these three hormones are not changed correlating with the week's growth of the rats.

Abstract:Intestinal adhesion can cause serious complications, so it has been a common problem after surgery. In order to solve this problem, investigating the formation mechanism of intestinal adhesion, as well as the preventive and therapeutic measures become a focus in surgery. As important carriers, animal models have played very important role in the research of formation mechanism, prevention and treatment of intestinal adhesion. In this article, the establishment and evaluation methods of animal models of intestinal adhesion will be reviewed.

Abstract:Laboratory animal experiments of pathogenic infection play more and more roles in animal medicine and biological product research. We surveyed the scientific papers involved with poultry infection experiments published in NCBI Pubmed from Jan.1^st, 2009 through April 29^th,2014, focused on the "humane endpoint" and euthanasia. Among the 247 reports, we found that the euthanasia methods adopted by the researchers were diverse, though almost all authors claimed that they complied with the Animal Experiment Care and Welfare Committees. Carbon dioxide inhalation and anesthetic injection are the most commonly used ways, while the reagents and doses are various. Moreover, the judging standards for experimental or humane end-point vary across the experiments, there are still no standard routes or doses when using anesthetics yet. The paper also compares the characteristics of reported euthanasia methods.