Abstract:Objective To explore the feasibility of establishing mouse model of leukemia by transplantion of human acute lymphoblastic leukemia (ALL) CD34⁺CD38- cells into immunodeficient (NOD/SCID) mice, and to study the capability of self-renewal and proliferation in the mice. MethodsThe CD34⁺CD38- cells were isolated from ALL patients and then identified, and the CD34-CD38⁺ cells were used as control. These cells were transplanted into sublethally irradiated NOD/SCID mice by tail-vein injection of 10⁴ cells per recipient. The changes of peripheral blood were monitored continuously, and histopathological examination of the bone marrow, spleen and liver was performed in each death or dying mouse. ResultsThe number of leukocytes in mouse peripheral blood was increased after inoculation of CD34⁺CD38- or CD34-CD38⁺ cells at 4 weeks, peaked at 8 weeks up to 15×10⁹～20×10⁹/L, and also increased the number of original and blast lymphocytes. There were mainly increased the number of the original and blast lymphocytes (40%) in the bone marrow, as well as the leukemic cells infiltrating in the spleen and liver of CD34⁺CD38- mice, which was more obvious than that of the control CD34-CD38⁺ mice. ConclusionsCD34⁺CD38- cells from ALL patients can be successfully transplanted into NOD/SCID mice to induce leukemia, indicating that the CD34⁺CD38- cells have the capability of self-renewal and proliferation. This model can be used as an important and useful tool for studies on leukemia-initiating cells. 

Abstract:ObjectiveTo investigate the behavioral changes induced by repeated electrical stimulation of the dura mater surrounding the superior sagittal sinus in conscious adult rats. MethodsTwenty-four male rats were randomly divided into 2 groups. The experimental group was given electrical stimulation (electric current 1 to 2 mA, frequency 20 Hz, sine wave, pulse width 250 μs, lasting 15 minutes, once a day) for 21 days. The changes in body weight, fluid consumption test and open-field test between the two groups in 21 days were compared to evaluate whether depression behavioral changes were induced. Results After 21 days of electrical stimulation, the body weight of the rats of the experimental group was significantly lower than that of the control group (P<0.05). In the experimental group, the number of square crossing and vertical rearing in open-field test was significantly decreased, sucrose solution consumption and preference were significantly reduced, and pure water consumption was significantly increased, all with a significant difference between the two groups (P<0.05). ConclusionsDepression behaviors have been successfully induced by repeated electrical stimulation to dura mater surrounding the superior sagittal sinus for 21 days in conscious rats.

Abstract:ObjectiveTo observe the influence of different formulas of chow and different ages of rats on the success rate and characteristic pathological changes in the establishment of a rat model of diabetes mellitus. MethodsTwenty-six 3-week-old SD rats were divided into 3 groups:control group 1 (group N1,6 rats), model group 1 (M1 group, 10 rats), and model group 2 (M2 group, 10 rats). Twenty-six 5-week-old SD rats were divided into 3 groups:control group 2 (group N2,6 rats), model group 3 (group M3,0 rats), and model group 4 (group M4,0 rats). The groups M1 and M3 were fed with the formula 1 high fat chow, and the groups M2 and M4 were fed with the formula 2 high fat chow. The rat models were given streptozotocin (STZ) injection (35 mg/kg) once intraperitoneally after feeding with high fat chow for 4 weeks. The levels of FBG, FIN, TG, TC were detected successively. ResultsThe FBG level of 5-week-old SD rats reached a stable state 2 weeks after STZ injection, and kept on a high level. The rats fed with the formula 2 of high fat chow showed significant increase of food-intake and body weight, and had successfully induced insulin resistance. Conclusions5-week-old SD rats fed with the formula 2 of high fat chow and STZ injection present major pathophysiological manifestations of type 2 diabetes mellitus, and can be used as a good model of type 2 diabetes mellitus in relevant research.

Abstract:Objective To investigate the effects of a Chinese traditional prescription PN-1 on behavioral and psychological symptoms in APP/PS1 transgenic mouse models of Alzheimer’s disease (AD). MethodsMouse models at 5 months of age were randomly divided into model group (vehicle), Aricept administration group (2 mg/kg), PN-1 at low (0.6 g/kg), middle (1.2 g/kg) and high (2.4 g/kg) doses. The wild-type littermates were chosen as normal control group (WT). The 16 mice (8 males and 8 females) in each group were given once every day by intragastric administration of PN-1, Aricept or water for 3 months. The body weight and the amount of water drinking and food intakes were measured before administration and every two weeks during administration. After the drug administration, a battery of behavioral tests including social interaction, open field, rota-rod and sucrose preference were performed. ResultsDuring the administration, no changes in body weight and food/water intake were detected in PN-1-treated mice, compared with the age-matched wild-type control and vehicle mice (P > 0.05). Compared with the mice in the vehicle group, social interaction test showed reduced attack, chasing and sniffing in the PN-1-treated groups (P < 0.05). The mice in the PN-1-treated groups showed decreased spontaneous motor activity (P < 0.05), highly mobile duration (P < 0.05), frequency to center zone (P < 0.05) and increased frequency of grooming in the open field test. Rota-rod test exhibited increased time spent on rod (P<0.05) for the PN-1-treated mice. In addition, increased sucrose consumption was also found in the PN-1-treated group, however, no statistically significant difference was found compared with the vehicle group (P>0.05). ConclusionsPN-1-treatment can ameliorate many non-cognitive behaviors and psychological symptoms in the APP/PS1 transgenic mouse models, including rescued hyperactive social interaction, spontaneous motor activity, improved ability to learn balance, and reduced anxiety, irritability and other psychological symptoms.

Abstract:ObjectiveTo establish a rat model of postpartum stress urinary incontinence (PPUI) through vaginal placement of urinary catheter balloon causing pelvic organ injury.MethodsAfter the vaginal catheter balloon dilation for 8 hours on 40 Sprague-Dawley female rats, 25 were randomly selected as experimental group and 5 normal rats of the same age as control group. Sneeze test was used on the day of the procedure and on weeks 1,2, 4 and 8. The animal models were evaluated through urodynamics as measured by leak point pressure (LPP), maximum bladder volume, pelvic floor muscle acetylcholine receptor content, comparison of serum lactate dehydrogenase (LDH) and creatine kinase (CK) according to the JSCC standards, pelvic floor muscle strength and RT-PCR assessment of caspase-3 expression. ResultsAfter 8 hours of vaginal balloon dilation, a rat model of PPUI was established with a success rate of 72.5% (29/40). In the model animals, LDH and CK were initially increased, peaked on day 7, and finally decreased toward normal values. The pelvic floor muscle contraction strength and pelvic pubococcygeus acetylcholine receptors were declined. There were significant differences in the values between experimental and control groups (P<0.05). Pubococcygeal muscle caspase-3 gene expression peaked on the day of the procedure (P<0.05) and decreased thereafter. ConclusionsRats subjected to vaginal balloon dilation for 8 hours display biochemical alterations in sera, activation of caspase-3, and decreased pelvic floor muscle contraction strength, similar to those observed in human PPUI patients. 

Abstract:Objective To analyze the population genetics of three closed rabbit colonies:Japanese white rabbits (JWR), Chinchilla rabbits (CR), and New Zealand rabbits (NZWR). MethodsTen microsatellite loci were selected and tested by Hardy-Weinberg equilibrium (HWE), and the values of the allele frequency, observed heterozygosity and expected heterozygosity, the F-statistic value and the genetic distance were counted. ResultsThe microsatellite locus 12L1E11 in CR population, the loci INRACCDDV0087 and INRACCDDV0203 in NZWR population, and the loci Sat12 and INRACCDDV0203 in JWR population showed evident deviation from that of the HWE (P<0.05), mostly resulted from heterozygote deficiency. All the other microsatellite loci Sat13, Sol4,6L1F0,7L1F1,2L4A1, and INRACCDDV0016 in the three populations consisted with that of the HWE. The average number of alleles was 5.9, and the range of allele frequencies was from 0 to 0.9060. The average observed heterozygosity was 0.6204 and the average expected heterozygosity was 0.6178. The average Fst value among populations was 0.0750. The observed lowest genetic distance was 0.1223 between JWR and CR,and the observed highest genetic distance was 0.1934 between CR and NZWR. ConclusionsThe genetic structures of the three closed rabbit colonies show genetic stability and homogeneity. The 10 microsatellite loci have high genetic polymorphisms. There is evident genetic differentiation among populations.

Abstract:Objective The polymorphism of microsatellite loci among four inbred strains of rats ( HFJ, MIJ, Lewis, F344) was analyzed using 24 pair primers.MethodsTo extract the genomic DNA of MIJ, HFJ, Lewis and F344 inbred rats by standard extraction procedure with phenol/chloroform. 24 microsatellite loci were selected for PCR amplification. Through native polyacrylamide gel electrophoresis and silver staining, microsatellite polymorphism was analyzed in the four strains of inbred rats. ResultsThe amplified products of 24 microsatellite loci among different individual rats of these 4 strains and the same strain rats displayed single allelic gene band. Among the 24 microsatellite loci, it displayed monomorphism between HFJ and MIJ rats. But HFJ and MIJ rats had significant deviations from Lewis and F344 rats. 14 microsatellite loci turned out to be polymorphic, and 10 microsatellite loci displayed monomorphism. ConclusionsThe results of this study suggest that MIJ and HFJ strains basically meet the requirement of inbred strain. The 14 screened loci can be used for genetic monitoring of these inbred rats.

Abstract:Objective To investigate the possible neurotropism, neuroinvasion and their mechanism in a Chinese-origin rhesus macaque infected with SIVmac251 showing neurological symptoms. MethodsEight healthy laboratory rhesus macaques were infected with SIVmac251-155p6N by intravenous injection. Among them 3 monkeys showed neurological symptoms to a different degree, and one of these 3 animals (R21755) developed severe neurological symptoms. This monkey was sacrificed and the virus was isolated from the basal ganglia of the brain, the plasma viral load was quantified by real-time RT-PCR, and T cell subsets were determined by flow cytometry. The brain lesions were examined by histopathology using HE staining. The RNAs were extracted from SIVmac251-155p6N, plasma and basal ganglia, the gp120 genes were amplified with single genome amplification, and the changes of N-glycosylation site in gp120 regions were analyzed. ResultsThe monkey R21755 presented AIDS encephalopathy-like symptoms after half a year post the viral infection. Pathological examination showed infiltration of macrophages and multinucleated giant cells in perivascular space, and degeneration and necrosis of neurons were observed in the brain tissue. The gp120 sequence analysis showed that the virus isolated from the brain was partly different from that of the other sequences of SIVmac251-155p6N and plasma virus of the monkey R21755. The variations were mainly in the V1/V4 regions and the loss of a N-glycosylation site in env C1. ConclusionsIt seems that the neuroinvasion and neurovirulence of SIVmac251 are significantly enhanced during the long-term adaption in vivo. The finding of this study provide a firm molecular basis for establishing a strain of SIVmac251 with neurotropism and neurovirulence. This would be very meaningful for investigating the role of SIV in neurologic disease and studies of AIDS neuropathy. 

Abstract:Objective To isolate and obtain the complete sequence of Trp1 and Trp2 genes from white hair black eyes(WHBE) rabbit iris, which represent the second and third members of the tyrosinase-related gene family, and to identify the sequences of protein TRP1 and TRP2.MethodsThe WHBE rabbit iris whole mRNA was reversely transcripted to cDNA. The cDNA was isolated from mouse, rat and human using homologous primers. The fragments and complete sequences of the cDNA were obtained by PCR and RACE. ResultsThe WHBE rabbit coding region was sequenced completely to yield 1604 bp full length and shared 87.9% identity with the human sequence and 82.7% identity with the mouse sequence. The Trp1 mature protein sequence had 513 amino acids and shared 89.8% identity with the human sequence and 86.6% identity with the mouse sequence. The WHBE rabbit Trp2 coding region was sequenced completely to yield 1554 bp full length and showed 83.2% identity with the human sequence and 81.9% identity with the mouse sequence. The WHBE rabbit Trp2 mature protein sequence had 494 amino acids and shared 84.2% identity with the human sequence and 84.4% identity with the mouse sequence. ConclusionsThe protein sequences of TRP1and TRP2 obtained from WHBE rabbit iris in this study show a high homology to TYR and show structural characteristics of the tyrosinase-related protein family. 

Abstract:Objective To explore the treatment and anti-oxidation mechanism of total alkaloids from Pinellia ternate (TAPT) in Parkinson’s disease (PD) rat models.MethodsThe rat model of Parkinson’s disease was established by injection of 6-hydroxydopamine (6-OHDA) into the rat brain. TAPT was given to the model rats to observe its preventive effect. The behavioral changes of the PD rats were tested with Morris water maze. Chemical assay was used to measure the superoxide dismutase (SOD) activity, malondialdehyde (MDA), hydrogen peroxide (H₂O₂) and glutathione (GSH) contents in the cerebral cortex and serum. ResultsCompared with the normal group, the escape latency of the PD rats was significantly longer (P<0.01), the number of platform crossing significantly reduced (P<0.01). After TAPT treatment, the escape latency of PD rats was significantly shorter (P<0.05, P<0.01), and the number of platform crossing was significantly increased (P<0.01). In the Parkinson model group, the MDA and H₂O₂ contents were increased, the SOD and GSH levels in serum and cerebral cortex were significantly decreased (both P<0.01), the MDA and H₂O₂ contents were significantly reduced (P<0.01), and the GSH and SOD levels in the cerebral cortex were significantly increased (P<0.01). The serum MDA levels in the low concentration and middle concentration TAPT treatment groups were not significantly changed (P> 0.05), but significantly decreased in the high concentration TAPT treatment group (P<0.01). The serum H₂O₂ in the treatment groups was significantly decreased (P<0.01), and serum GSH contents in the treatment groups significantly increased (P<0.01). ConclusionsTAPT treatment show improving effect on learning and memory in PD rats, and may play a role against the degeneration in the rat nervous system, possibly through changes in the cortical and serum SOD, GSH levels, while inhibit of MDA and H₂O₂ generated in the rats with Parkinson''s disease. 

Abstract:ObjectiveTo study the pathological and immunological changes of chronic dermatitis in KM mutant mice. MethodThe changes of external characteristics were observed by stereomicroscopy and optical microscopy, and the immune cells and inflammatory factors were observed using HE staining, immunohistochemistry and toluidine blue staining in 3-, 6-month old wild-type and KM mutant mice. ResultsThe skin changes showed oligotrichia, scurf, rhicnosis and the histopathology showed epidermal cell necrosis, epithelial hyperkeratosis or parakeratosis, hypergranulosis, basal cell layer edema, dilatation of blood vessels in the superficial layer of dermis and infiltration of inflammation cells in the connective tissue; increase of cutaneous CD3+ and CD4⁺T cells, macrophages and mast cells; and increase of inflammatory factors IL-6, IL-22, TNF-α, and IFN-γ in the 6-month old KM mutant mice and more severe in the 3-month old KM mutant mice . ConclusionsThe skin tissue of KM mutant mice shows chronic inflammatory changes in pathology and cell-molecular changes similar to that in human chronic inflammatory skin diseases. KM spontaneous mutant mice may become a novel animal model in research of of chronic dermatitis .

Abstract:Objective To investigate the effects of naringin and the combined application of naringin with evodiamine on cognitive ability in APP^swe/PSΔE9 transgenic mice, and to compare the difference of neuroprotective effects between the applications of these two drugs alone or in combination, and to determine whether there is synergistic or additive effect of the combination of naringin and evodiamine. Methods ① APP^swe/PSΔE9 transgenic mice aged 3 months and 5 months were randomly divided into different groups, 12 in each group. The mice performed Morris water maze test after administration of the drugs for 4 weeks and 16 weeks, respectively. The drugs were mixed in the diet singly or in combination according to the grouping protocol. One group was used as vehicle group and one group was used as Aricept group. The littermates receiving placebo were used as normal control. ②The senile plaques were detected by Thioflavin-S staining and visualized directly by fluorescence microscopy. Results ① Compared with the APP^swe/PSΔE9 transgenic mice, naringin administrated alone for 4 weeks and 16 weeks showed a significantly decreased latency by 22% and 32%, respectively; and evodiamine administrated alone for 4 weeks and 16 weeks showed a decreased latency by 33% and 10%, respectively. ② The senile plaques were decreased by 26% in the hippocampus of mice after 16 weeks naringin administration. ③ The combined application of naringin with evodiamine showed the decreased latency by 47%, compared with the APP^swe/PSΔE9 transgenic mice, however, when compared with naringin alone, the combined application showed no more decreased latency. ConclusionsEither naringin or evodiamine can improve the cognitive deficits in APP^swe/PSΔE9 transgenic mice. Long-term effect of naringin is obviously better than its short-term effect. However, on the contrary, short-time use of evodiamine is effective but its long-time effect is decreased. The combined application of naringin with evodiamine can obviously improve the cognitive ability in APP^swe/PSΔE9 transgenic mice, however, it shows no synergetic protective effect compared with the use of naring in alone. 

Abstract:Objective To establish a rat model of progressive mesangial proliferative glomerulonephritis with vasculopathy by high fat diet feeding. MethodsMale Wistar rats were subjected to unilateral nephrectomy and the surviving rats were randomly assigned into 4 groups:high fat diet group, nephritis group, nephritis fed with high fat diet group, and unilateral nephritis rats severed as controls. OX7(100 mg/kg)antibody was firstly injected into the tail vein of rats of the nephritis group and nephritis fed high fat diet group at 3 days after nephrectomy, and then injected once per week for 3 times. The rats of nephritis group and high fat diet group received saline injection. Vitamin D3 was injected intraperitoneally in a dose of 600 000 U/kg to the rats at the second day after OX7 antibody injection, once per week for 4 weeks and fed with hgh fat diet. The general status was recorded and the body weight, systolic blood pressure, proteinuria, serum total protein, serum albumin, triglyceride, cholesterol,serum creatinine and serum calcium were measured. The renal function was assessed and renal alterations were observed by histopathology. ResultsIntrarenal arterial wall thickening, arterial stenosis and proliferation of fibrous tissue were observed in the rats of nephritis and high fat diet group at week 8. At week 10, the intrarenal arterial wall was slightly thickened in the rats of nephritis and high fat diet group. The scores of glomerular, tubulointerstitial and vascular histological damages in the rats of nephritis with high fat diet group were significantly higher than those in the rats of nephritis group and high fat diet group (P<0.05). ConclusionsAnimal model of progressive mesangial proliferative glomerulonephritis with vasculopathy can be successfully established by high fat diet plus intraperitoneal injection of vitamine D3 in chronic anti-Thy1 nephritis rats.

Abstract:Objective To investigate the effects of circadian rhythm changes on the expression of opsin photopigment melanopsin in the retinsa. Methods14-day old C57BL/6J mice were randomly divided into experimental group and control group. The experimental group was maintained in continuous light. The control group was maintained in a 12:12 h light:dark cycle. Fluorescence immunocytochemistry and RT-PCR assays were used to detect the expressions of melanopsin at 1 week and 8 weeks after the light exposure. ResultsThe results of fluorescence immunocytochemistry showed that opsin melanopsin mainly expressed in the retinal ganglion cell layer and little in the inner nuclear layer. The total amount of melanopsin cells in the experimental group were less than that in the control group 1 week later. RT-PCR indicated that the total amount of melanopsin mRNA in the experimental group was less than that in the control group (P<0.01). ConclusionsConstant lighting down-regulates the expression of melanopsin. Melanopsin-expression ganglion cells are photosensitive and may play an important role in the maintenance of normal circadian rhythm.

Abstract:ObjectiveTo observe the effect of tacrolimus at different concentrations on the blood glucose in rats. Methods 30 male SD rats, weighing about 80 g, were randomly divided into three groups. Group A was given tacrolimus (Astellas Ireland Co., Ltd) 2 mg/kg per day and Wu Zhi soft capsules containing schizandrin A and other components (Sichuan Guangda Pharmaceutical Co., Ltd.) 0.2 / kg per day for 5 months. Group B was given tacrolimus 2 mg/kg per day for 5 months. Group C was used as control and received daily saline solution. The three groups of rats were administered orally in the fasting state. The tacrolimus plasma concentration and fasting blood glucose were monitored monthly. Results Blood concentration of tacrolimus of group A was significantly higher than that in the group B ( P <0.01). In the first two months of drug administration, there was no significant difference in fasting blood glucose among the three groups (P> 0.05). The Groups A and B had significantly higher blood glucose than in the group C three months after treatment (P <0.01) . In the last two months the blood glucose of the groups A and B continued to rise, and the blood glucose in group A was significantly higher than that in group B (P <0.01). ConclusionsTacrolimus causes elevated blood glucose by reducing insulin secretion and increasing insulin resistance. This glycemic effect is time-dependent and dose-dependent.

Abstract:ObjectiveTo compare the susceptibility of different mice strains in different immune status to H5N1 virus,and investigate the possible mechanism. MethodsGerm free BALB/c mice,SPF BALB/c mice,SCID mice and nude mice were challenged with 50 μL 10 TCID50 H5N1 virus, then the body weight and survival were observed daily. In addition,the virus distribution in tissues,the cytokines in the lung,and pathological changes of the lung were also monitored. ResultsThe four different strains of mice could be infected by H5N1 virus successfully. The germ free mice were the lowest susceptible animals,with lowest virus replication in GF mice tissues and lowest cytokine level,including TNF-α,IL-12,MIP2,GATA3,IFN-α/β/γ,especially at the 7^th day. ConclusionsGerm free mouse is the lowest susceptible animal to H5N1 among the four mouse strains:germ-free BALB/c,SPF BALB/c,nude and SCID mice,due to low level of virus replication and low cytokine expression.

Abstract:ObjectiveTo observe the expression of glutamine fructose-6-phosphate aminotransferase 1 (Gfat1) in the lung tissue of type 2 diabetic rats.MethodsSprague-Dawley rats were randomly divided into control group (n=18) and diabetic model group (n=28). Rats in the model group were fed with high-fat diet for 2 months, and then were injected intraperitoneally streptozotocin (STZ) at a dose of 15 mg/kg to generate diabetic models. Fasting blood glucose and body weight were recorded. The expression of Gfat1 mRNA in the lung tissue was detected by RT-PCR on the 2nd, 4th and sixth week after diabetes modeling, respectively. ResultsThe body weight of rats in the model group increased rapidly. Compared with that of the control group, body weight was significantly increased at the 28th, 42nd, 56th and 70th days from the modeling started (P<0.05). Fasting blood glucose (FBG) in the rats of model group after STZ injection was significantly higher (≥10.0),[JP+1] compared with that in the control group (P<0.01). Gfat1 expression in the rats of model group on the second week of diabetes modeling was lower than that in the control group, but the difference was not significant (P>0.05). Gfat1 expression of rats in the model group on the fourth week of diabetes modeling was higher than that in the control group, with a significant difference (P<0.05). Gfat1 expression in the rats of model group on the sixth week of diabetes modeling was higher than that in the control group, but the difference was not significant (P>0.05). ConclusionsType 2 diabetic rat model can be successfully established by feeding high-fat diet with intraperitoneal injection of streptozotocin. The level of Gfat1 expression is changed in the lung tissue at different periods of diabetes modeling.

Abstract:Besides commonly used rodents, there are a variety of experimental animals involved in the study of sleep, including cat, drosophila, monkey, zebrafish and so on. Rodents are used widely in sleep studies, because it is easy to make sleep models on rodents by physical and chemical methods. Among them, transgenic mice are mainly used for drug development and of sleep mechanism study. Drosophila and zebrafish are usually applied to genetic studies of sleep. Using cats for sleep study has a long history, and they are now often used to quantify in vivo endogenous neurotransmitters. Monkeys have a similar sleep structure to humans, so that they are widely used to study the relationship between sleep and neurobiology, behavioral pharmacology and so on. Animals are selected for experiments on the basis of research purpose, because each kind of animals has its own unique physiological characteristics.

Abstract:Compared with the standard cages, laboratory mice and rats prefer the cages enriched with nesting materials. Nesting materials provided in conventional mouse or rat cages can benefit the rodent''s wellbeing, and have been widely used in husbandry and management of laboratory mice and rats for improvement of welfare. Here, we review the background, application, and effect of nesting materials on environmental enrichment and welfare improvement for laboratory mice and rats. This review also provides a clue to improvement of laboratory animal welfare in our routine work. 