Abstract:ObjectiveTo separate and identify the characteristics of testicular metal-binding proteins (TMBPs) induced by exhaustive swimming in rats. MethodsEight male SD rats were subjected to a bout of exhaustive swimming (8 for control) and the testes and liver were taken out 6 hours after the exercise. TMBPs and hepatic metallothionein (MT) were determined by cadmium-saturation assay. The extracts from the testes and liver by cadmium-saturation method were separated by tricine-SDS-PAGE and TMBPs protein bands were identified by LC-MS-MS. ResultsThe results of cadmium-saturation assay showed that the TMBPs content was significantly increased in the exhaustive swimming rats [(113.71±11.72)nmol Cd/g testis] than that in the controls [(87.14±12.72)nmol Cd/g testis] (P<0.01), and the hepatic MT was (64.70±14.89)μg/g, significantly higher than the (7.32±3.31)μg/g in the controls (P<0.001). Ubiquitin, superoxide dismutase [Cu-Zn](Cu-Zn-SOD), casein-like phosphoprotein (CLPH) were identified in TMBPs, and MT was also included. ConclusionsTMBPs are mainly composed of ubiquitin, SOD and CLPH, with characteristics of metal binding, heat resistance and induction. The cadmium-saturation assay is not a specific method for MT determination.

Abstract:ObjectiveThe object of this study was to compare the immunological characteristics of brown Norway (BN) rats and guinea pigs challenged with ovalbumin (OVA). MethodsAnimals were actively sensitized to OVA or sham sensitized, and 10 days later lymphocytes function in the spleen or bone marrow, activity of peritoneal mast cells and changes of calcium concentration in the peritoneal mast cells were tested. ResultsBoth phytohemagglutinin-P (PHA-P) and lipopolysaccharide (LPS)-stimulated lymphocyte proliferation in the spleen and bone marrow, and the bone marrow lymphocyte activity in the OVA-sensitized rat group were significantly higher than that of the guinea pig group (P<0.05). Annexin-V was used as the biomarker in the process of degranulation of mast cells in the present study. There was an increase of annexin-V binding mast cells in the OVA-sensitized animals, and the amount of mast cells in the OVA-sensitized rat group was twice more increased than that in the guinea pig group. The calcium concentration was also increased in the OVA-sensitized animals, but showed only a significant difference in the calcium concentrations in the mast cells of guinea pigs before and after OVA-sensitization (P<0.01). ConclusionsThe results of this study indicate that repeated intraperitoneal OVA challenges induce immunological responses differently in the BN rats and guinea pigs. The activity of bone marrow lymphocytes in the OVA-sensitized BN rats is considerably higher than that in the sensitized guinea pigs, but the calcium concentration in the mast cells of sensitized guinea pigs is higher than that of BN rats. An appropriate animal model should be selected according to the relevant immunological characteristics and actual demands of the experiment.

Abstract:ObjectiveTo develop a technique for establishing a rat model for end-to-side anastomosis of the common carotid artery (CCA). MethodsA surgical operation was performed to create an end-to-side anastomosis between the right and left CCA passing through the tunnel between the anterior cervicalmuscle group and the trachea in rats. ResultsAll the 10 Sprague-Dawley rats which underwent this procedure, survived the operation without postoperative neurological sequel. The success rate was 100%. The mean number of sutures required to complete the anastomosis was eight. The time spent to perform the anastomosis was (35±5)min. The anastomoses were almost 100% patent immediately and 3 months after the procedure. HE staining showed good anastomotic healing. ConclusionsA rat model for end-to-side anastomosis of the common carotid artery (CCA) has been successfully established. Careful operational micromanipulation and the protection of the small branches of nerves and blood vessels are the two key points for successful anastomosis. It may serve as a useful model for suture training in microvascular surgery.

Abstract:ObjectiveTo explore whether reared baby or not influences the social behavior of female Macaca fascicularis raised in free enclosure. MethodsFrom May 2010, we randomly selected 40 monkey samples of the observation-available subjects from the rear base of Jiangsu Province Suzhou Zhongke Experimental Animals Co. Ltd. The formal observation was conducted from Oct 2010. Of the 40 female monkeys, 11 subjects reared baby and 29 subjects did not. Focal observation method using high resolution digital camera was employed by three trained observers to record the behavior of target subjects, 7 phases per day, 30 min per phase, and each subject was observed for 4 days, with a totally effective observation time 560 hours. The recorded video clips were randomly delivered to the Noldus analysis system with an analysis reliability kappa >0.8, and transferred into behavior data. ResultsThe subjects, who adopted baby, just hold the baby in most of time, more social amicable behavior, for instance, mutual grooming for others or being groomed by other companion, higher social rank, namely to share the rest place with monkey king on the board (which was hung across over the room) for longer time; more communication behavior, lip smacking; and more locomotive behavior and mating behavior with male subjects than those female monkeys not reared baby. Notwithstanding, the subjects who did not rear baby spent more time on embrace behavior with other companions. Otherwise, there were not significant differences on ingestive, rutting, rest, conflict, vigilance and miscellaneous behaviors. ConclusionThe reared baby has influence on the social behavior of female Macaca fascicularis.

Abstract:ObjectiveTo explore a more effective, stable method to establish animal model of coronary artery stenosis assisted by thoracoscopy. MethodsThe proximal left anterior descending branch of the left coronary artery (LAD) in Bama miniature swine was ligated permanently using a silk suture by thoracoscopy, and induced LAD stenosis to different degrees. Selective coronary artery angiography was performed postoperatively to evaluate the degree of stenosis by quantitative coronary angiography. ResultsAmong the 20 swines, coronary artery stenosis models were successfully constructed in 18 swines. Ventricular fibrillation occurred in 2 swines during the procedure and they died 8 hours and 48 hours after effective defibrillation, respectively. The results of selective coronary angiography showed that there were LAD stenosis degree 20~50% in 7, stenosis degree 50~70% in 6, and stenosis degree 70~90% in 5 swines. The success rate of operation was 100%, and the success rate of model construction was 90%. ConclusionsFull-thoracoscopic operation can be successfully used to construct animal models of coronary artery stenosis to different degrees with less injury than open thoracic surgery.

Abstract:ObjectiveTo establish and evaluate an acute exercise-induced fatigue animal model. Methods Twenty-four male Wistar rats ( 8-week-old) were used in this experiment. Multilevel increasing load (3 levels) of exercise scheme on the treadmill (0° slope) was applied to establish an animal model of one single bout of exhaustive exercise. Peripheral blood samples were taken from the tail vein at different time points (before exercise, 30 min, 90 min and exhaustion during exercise, and 30 min and 90 min after exercise), andblood concentrations of GLU, LD, BU, MDA and activities of CK and SOD were detected. ResultsThere were dynamic changes in the behavior and exercise ability, blood metabolites and energy material during the exhaustive exercise in the rats. There were significant increases in the levels of LD, BU, MDA (P<0.01) and activity of CK (P<0.05) in peripheral blood compared with those at rest. The concentration of GLU and activity of SOD in peripheral blood were significantly decreased, compared with those at rest (P<0.01, P<0.05). The characteristic changes of all indexes indicated that fatigue occurred. ConclusionsA rat model of one single bout of exhaustive exercise on treadmill has been successfully established, and the characteristics of changes and regulation of indexes at different stages (generation, development and recovery of fatigue) are objectively evaluated. This rat model is suitable for the research on exercise-induced fatigue and its mechanisms.

Abstract:ObjectiveTo explore the significance of microstructure of cochlea and some physiological indexes in evaluation of the rat models of iron-deficient nephrasthenia deafness. MethodsOne hundred and twenty healthy Wister rats at age of 1-2 weeks and weighing 30-32 g, without ear disease and within the normal range of ABR threshold, were included in this study. The rats were randomly divided into experimental group (n=80) and control group (n=40). The male and female rats were fed separately. Iron-deficient diet was fed to the experimental rats and normal diet was fed to the normal rats, both for 12 weeks. According to the occurrence of nephrasthenia syndrome and ABR ≥15dB in at least one ear, 22 rats from the experimental group were diagnosed as nephrasthenia deafness, among which 20 deaf rats were taken as the experimental group, and 20 normal rats as the control group. ABR threshold was tested, hemoglobin and serum iron content were measured, and histological changes of the cholea such as stria vascularis and spiral organ were examined in all the rats of both groups. ResultThe ABR was (30±5) dB in the experimental group and (10±5)dB in the normal group; the hemoglobin was 11.80 g/L in the experimental group and 4.5 g/L in the normal group; the serum iron was 45.9 μmol/L in the experimental group and 22.23 μmol/L in the normal group, all showing a significant difference between the two groups. In the experimental rat cochleae, the blood vessels of stria vascularis were apparently decreased, and the trichobothria of hair cells were lost or lodging. ConclusionsThere are quite stable changes of the indexes such as hemoglobin and serum iron, ABR threshold, and the vascular changes in the cochlea stria vascularis and spiral organ in the rat models of iron-deficient nephrasthenia deafness, therefore, they may be good indicators for evaluation of this disease.

Abstract:ObjectiveTo explore the optimal methods of isolation and culture of hepatocytes from grass carp and serve the studies on nutritional metabolism and injury mechanisms in carp hepatocytes. MethodsProliferation of hepatocytes was tested by MTT assay. Function of THE hepatocytes was examined by measuring the levels of lactic acid dehydrogenase (LDH), albumin (ALB), and urea nitrogen (BUN) in the supernatant at different times, respectively. Results Using 0.25% warm trypsin digestion for 20 minutes each time, and to collect the hepatocytes step by step, the results of trypan blue test and blood cell counting chamber method showed that more than 99% of viable hepatocytes were obtained. ConclusionThe cells were cultivated in M199 medium supplemented with 10% fetal bovine serum and 10μg/mL insulin, at 7×10⁶ cell/mL concentration under the temperature of 28℃, 4.5% CO₂ for a long term. Primary culture of carp hepatocytes can be successfully obtained by the following procedure:cells cultured in M199 medium supplemented with 10% fetal bovine serum and 10 μg/mL insulin, at 7×10⁶ cell/mL concentration, under the temperature of 27℃ and 4.5% CO₂.

Abstract:ObjectiveTo isolate simian adenovirus (SAdV) in vitro. MethodsSimian adenovirus in fecal specimens of monkeys were detected by PCR. SAdV-positive samples were inoculated into BSC-1 cells. The cells were blindly passaged for three times to observe cytopathic effect in cell cultures. The presence of SAdV was confirmed by PCR-sequencing and electron microscopy. ResultsA strain of monkey adenovirus was isolated form Macaca fascicularis in Guangdong province, hence it was named as GD strain. The cytopathic effect was obvious 24 hours after infection. Viral particle-like adenovirus were observed by electron microscopy. The GD strain had high nucleotide sequence homology with HAdV-G. Pol and hexon genes of the isolated strain had 96% and 83% nucleotide sequence homology with SAdV-1, respectively. ConclusionsThe successful isolation of SAdV can facilitate the research for its biology, pathogenesis and laboratory diagnostics, and make it possible to construct SAdV gene transfer vector that benefits development of alternative carrier.

Abstract:Objective To obtain X-ray computed tomography (CT) imaging of abdominal and pelvic organs of healthy adult rhesus monkeys. MethodsSix conventional laboratory rhesus monkeys, 3 male and 3 female, in the age of 5-8 years were included in this study. The monkeys were placed on the CT table-board in supine position and were scanned to get CT images of the abdominal and pelvic organs by enhanced scan techniques. Volumetric data were collected and reconstruction of the original data was conducted. Meaningful anatomical scanning images were observed, identified and selected:7 images of the abdominal structures and 10 pelvic organs (5 of males and 5 of females), and each layer of the major visceral organs were labeled. ResultOn the CT images, the subtle changes of most organs and tissues could be distinguished, and the interface of relatively large organs and blood vessels was clear. But the interface of smaller organs, blood vessels, nerves and muscles, etc. was not distinct. ConclusionsCT images of normal abdominal and pelvic structures of healthy adult rhesus monkeys has been obtained in this study. It provides valuable imaging basis for further studies on disease diagnosis and scientific experiments. 

Abstract:Objective To obtain microsatellite DNA sequence of Microtus fortis from its BAC library. MethodsTo screen the microsatellite DNA sequence with nonradioactive analysis method—colony hybridization and enrichments by magnetic beads. ResultsTwenty positive clones with the strongest signals were screened out of 136 BAC clones through hybridization of digoxin-labeled oligonucleotide (CA)₂₀. Then these positive BAC clones were devoted to construct the subclone libraries separately. 220 microsatellite sequences were isolated from 400 positive subclones identified by PCR. According to those microsatellite sequences with more copy numbers and complete flanking sequence,74 pairs of primers were designed. As a result, 35 pairs of microsatellite primers got clear bands, of which 16 pairs had polymorphism. ConclusionsMicrosatellite DNA sequences have been screened successfully and efficiently from BAC library of Microtus fortis. The sequences and positive BAC clones may contribute to further research of their location. 

Abstract:Objective To observe the suppressive effect of 3,5-dichlorophenylbiguanide on pulmonary metastasis of melanoma in vivo. MethodsHuman melanoma cell line M21 cells were injected into the tail vein of nude mice (defined as day 1). The treatment procedure was carried out at days 5,9, 3,7, and 21 by tail vein injection of 3,5-dichlorophenylbiguanide at a dose of 6 mg/kg, 12 mg/kg or 24mg/kg to the nude mice of the experimental groups. In addition, positive control, negative control, and vehicle control groups were set up. The mice were sacrificed at day 50. The lungs were removed and fixed in Bouin’s fixative solution for 24 h. Metastasis scores were figured out by a formula according to the diameters and numbers of white metastatic nodules on the lung surface. ResultsThe 3,5-dichlorophenylbiguanide treatment reduced the metastasis scores to 55.25±13.60 (6 mg/kg), 35.13±17.36 (12 mg/kg) and 12.83±11.44 (24 mg/kg), significantly lower than that of the vehicle control group (P<0.05). The cRGDfV treatment reduced metastasis score to 11.50±10.44, significantly lower than that of the vehicle control group (P<0.05). The metastasis score of the negative control IH1066 treatment group was 88.50±17.21, and that of the untreated group was 88.88±11.29, both with a non-significant difference compared with that of the vehicle control group (82.50±17.72) (P>0.05). Conclusion3,5-dichlorophenylbiguanide shows a suppressive effect on the pulmonary metastasis of melanoma in mice in a dose-dependent manner.

Abstract:Objective To establish a PiggyBac transposase-expressing transgenic mouse model for study of the genetic modification mediated by transposon in mice. MethodsPiggyBac transposase gene was driven by CMV promoter and transgenic mice were created by the microinjection. The gene type of transgenic line was identified by PCR and the gene expression level in testis was determined by RT-PCR. The activity of the PiggyBac transposase was detected with the transposition efficiency by Southern blot in the mice crossed with transposon donor transgenic mice. ResultsSeven lines of transposase transgenic mice were obtained by microinjection. One mouse line with relatively higher expression level of transposase in the testis was obtained and the transposase induced the transposition of the transposon donor DNA fragment in the mouse genome. ConclusionA PiggyBac transposase-expressing transgenic mouse model is successfully established. This model will greatly contribute to the research of the genetic modification mediated by transposon in mice. 

Abstract:Objective To explore the effects of raloxifene on pituitary gland in rats. MethodsOvariectomized Wistar female rats were implanted with silastic tubes containing raloxifene, 17-β estradiol or blank silastic tube alone, respectively. After 8 weeks, all the rats were weighed and sacrifieced. The pituitary glands were collected and weighed. Serum prolactin level (PRL) was tested by solid phase-sandwich method. ResultsThere was no significant difference between the pituitary weights of the raloxifene and control groups. However, the pituitary weight of the estrogen group was significantly higher than that of raloxifene and control groups. (both P <0.05). The serum PRL level in the estrogen group was significantly higher than that in the raloxifene and control groups (both P<0.05). The serum PRL level of the raloxifene group was significantly higher than that of the control group (P<0.05). The serum PRL level in the raloxifene group was significantly lower than that of the estrogen group (P<0.05). Histological examination showed normal pituitary morphology in the raloxifene and control groups, but characteristic changes of prolactinoma were seen in the pituitary gland of estrogen group. ConclusionRaloxifene has a certain impact on rat pituitary gland, but does not induce prolactinoma.

Abstract:Objective The present study aimed to establish a chicken model of fatty liver hemorrhagic syndrome (FLHS) induced by a combination of injection of estrogen and feeding of high fat diet, and to evaluate the clinical symptoms, blood lipid levels, liver function and pathomorphology of these chicken models. MethodsThree hundred twenty 14 day-old Cyan-Shank Partridge chickens were randomly divided into 4 treatment groups (4 replications of 20 birds in each group):Control (C), high-fat diet (HF), estrogen injection (E), and high fat diet plus estrogen injection (HF+E) groups. The experimental period lasted from 15 to 42 days. Two 14-day periods were used to assess different factors. The clinical symptoms of the chickens were monitored each day over the experimental period. Abdominal fat weight, pathological changes, serum and liver parameters were measured at the end of each 14-d periods. ResultsFLHS occurred in the group HF+E at 14^th day whereas FLHS didn’t occur in the control group till 28^th day. The group HF+E showed obvious clinical symptoms such as mouth breathing, excessive sleepiness and abdominal sag in some birds 10 days later. Excessive fat deposition in the abdominal cavity and swollen, brown-yellow, friable, blunt and hemorrhagic liver were observed on day 14 and 28. Slight hepatocyte steatosis and small lipid vacuoles in cytoplasm were observed on day 14. A large number of hepatocytes displayed distinct pathological changes such as histological disarrangement, swelling and large lipid vacuoles in cytoplasm on day 28. The FLHS incidence was higher at 28^th day compared with that on the 14^th day. The serum triglyceride and total cholesterol, the relative weight of liver and abdominal fat, liver fat rate and liver hemorrhage score (LHS) were increased significantly compared with those in the control group on day 14 and 28 ( P<0. 05, P<0.01). The clinical symptoms, anatomic characteristics, pathological changes, blood biochemical indices and other findings in the group E and HF were similar to those in the group HF+E, but to a less severe degree and the time of occurrence was later than that in the group HF+E. ConclusionThe results of this study suggest that high fat diet combined with estrogen administration can successfully establish model of fatty liver hemorrhagic syndrome in immature chickens within 28 days.

Abstract:ObjectiveTo establish a rabbit model of chronic suppurative osteomyelitis of the mandible. Methods Twenty-two healthy New Zealand rabbits were used in this study. 0.1 mL 5% sodium morrhuate and 0.1 mL 5.0×10⁸CFU/mL suspension of Staphylococcus aureus were injected into the bone window in the lateral side of mandible body near the middle joint of the mandible. Clinical, cone beam CT (CBCT), bacteriological and histopathological examinations of the mandibles were performed at 16 weeks after infection.ResultsAll rabbits showed poor mental health, and fistulae and purulent secretion were seen in the early period after infection. Dual-energy X-ray examination showed no significant changes of the bone mineral density (BMD) in the surgical area (P >0.05). CBCT detected a blurred edge of the bone defects and signs of bone destruction. Staphylococcus aureus growth was detected in the surgical area. Pathological examination showed infiltration of lymphocytes, neutrophils and plasma cells, and sequestration. ConclusionsA model of chronic osteomyelitis can be successfully established by injection of sodium morrhuate and Staphylococcus aureus suspension into the bone window in the lateral side of mandible body near to the middle joint in rabbits. 

Abstract:Objective The aim of this study was to isolate, culture, and characterize the skin fibroblasts from Microtus fortis. MethodsThe primary skin fibroblasts were isolated from Microtus fortis at different ages by trypsin digestion and tissue adherence methods. The growth behavior and the characteristics of Microtus fortis skin fibroblasts(MfSF)cultured in DMEM and 1640 culture medium were studied, respectively. ResultsThe number of cells isolated from 1-3-day old animals was more profit than that from 5-day-old Microtus fortis, isolated by trypsin digestion, and the cell purity was high. The MfSF grew well and attached to the bottle wall with a high purity and good ability of proliferation in the primary passage in DMEM or 1640 culture medium, and the time of confluence was 6-7 days, but the growth became gradually poor in the subsequent passages. HE staining showed that the MfSF had an oval nucleus. It was unsuccessful to isolate and culture MfSF by tissue adherence method in this study. ConclusionA successful and efficient method of isolation and culture of skin fibroblasts from Microtus fortis has been established. It provides a useful method and protocol to obtain MfSF to serve further studies. 

Abstract:This article reviews the role of E-cadherin expression in the regulation of tumor metastasis. Recently,increasing studies on the regulation of E-cadherin in tumor metastasis have made considerable progress, especially on gene modification and transcriptional silence. The latest studies indicate that differential expression of microRNAs might be crucial in promoting metastasis and invasion via affecting transcription and expression of E-cadherin,which may be beneficial to develop novel diagnostic and therapeutic approach for metastasis. Clarifying the sophisticated regulation is not only of theoretical significance in deepening the molecular mechanism of cancer metastasis, but also of practical value at establishing normalized diagnostic and therapeutic systems and designing novel targeted drugs.

Abstract:Because of the similarities of progressive anti-thy glomerulonephritis to human IgA nephropathy and other mesangial proliferative glomerulonephritis, animal models of progressive anti-thy1 glomerulonephritis are used to study the mechanism and new treatment approaches of chronic kidney diseases. The establishment methods, pathogenetic mechanism and treatment of progressive anti-thy1 glomerulonephritis models are summarized and discussed in this review.