Effects of different types of drinking water on growth and fecal flora of mice / 中国比较医学杂志

Objective To explore the effects of different types of drinking water on the growth and fecal flora of mice.Methods Specific pathogen-free NIH mice were randomly divided into five groups,32 mice each group,with half males and half females in each group.The group were given either purified water(control group),acidified water,alkalized water,weakly acidic water or solid water.Diet and body weight were monitored continuously for 20 days.After the experiment,animal fecal samples were collected,and the V3-V4 region was amplified with bacterial 16S rDNA universal primers.An Illumina Miseq high-throughput sequencing platform was used for high-throughput sequencing,and microbial community,α diversity and β diversity were analyzed by bioinformatics method.Results The body weight of female mice given different pH values of weakly acidic water was higher,while the weight of the other groups was lower,than that of the control group(P＞0.05).The body weight of male mice in the acidified water group was higher,while that of other groups was lower,than that of control group,but there was no statistical difference between the groups(P＞0.05).The body weights of male and female mice in the solid water group were lower than those in the control group(P＜0.05).The food and water intake of the female animals in the alkaline water group and the water intake of female animals in the solid water group were lower than those of the other groups.OTU clustering analysis showed that the data volume of the sequencing was reasonable,and the fecal flora species of NIH mice were divided into five phyla,among which Bacteroides and Firmicutes were dominant.Unclassified Pseudopurpuromonas,Lactobacillus and Alistipes were the main genera.There were differences in fecal flora abundance and diversity among the mice given the five drinking water types.α analysis showed that the acidified water group had the highest flora abundance and diversity,while the solid water group had the lowest flora diversity.β analysis showed that the fecal flora composition in the solid water group was the closest to that of the control group,followed by the alkalized water group,acidified water group and weakly acidic water group.Conclusions Through an exploration of the effects of consuming different forms of water,this study revealed that solid water consumption had the greatest effects on body weight,feed intake,water consumption,and fecal flora of mice.The abundance and diversity of fecal flora in mice were affected by different pH values of drinking water,especially acidified water.

Human-centered understanding of laboratory animal welfare and ethics / 中国比较医学杂志

This review clarifies that laboratory animal welfare and ethics in life science research should be human-centered,positive and responsible through the understanding of the connotation of laboratory animal welfare.

Restoration of FMRP expression in adult V1 neurons rescues visual deficits in a mouse model of fragile X syndrome

Many people affected by fragile X syndrome (FXS) and autism spectrum disorders have sensory processing deficits, such as hypersensitivity to auditory, tactile, and visual stimuli. Like FXS in humans, loss of Fmr1 in rodents also cause sensory, behavioral, and cognitive deficits. However, the neural mechanisms underlying sensory impairment, especially vision impairment, remain unclear. It remains elusive whether the visual processing deficits originate from corrupted inputs, impaired perception in the primary sensory cortex, or altered integration in the higher cortex, and there is no effective treatment. In this study, we used a genetic knockout mouse model (Fmr1KO), in vivo imaging, and behavioral measurements to show that the loss of Fmr1 impaired signal processing in the primary visual cortex (V1). Specifically, Fmr1KO mice showed enhanced responses to low-intensity stimuli but normal responses to high-intensity stimuli. This abnormality was accompanied by enhancements in local network connectivity in V1 microcircuits and increased dendritic complexity of V1 neurons. These effects were ameliorated by the acute application of GABAA receptor activators, which enhanced the activity of inhibitory neurons, or by reintroducing Fmr1 gene expression in knockout V1 neurons in both juvenile and young-adult mice. Overall, V1 plays an important role in the visual abnormalities of Fmr1KO mice and it could be possible to rescue the sensory disturbances in developed FXS and autism patients.

Involvement of FKBP51 in mitochondrial damage induced by high fat diet feeding / 中国比较医学杂志

Objective To investigate the effect of high fat diet feeding on mitochondrial structure and function. Methods Male C57BL/6J mice at the age of 4 weeks were used in this study. After 6 weeks of regular diet(RD)or high-fat diet(HF)feeding, the high fat-induced obesity phenotype was confirmed by body weight measurement and liver histopathology. RNA was isolated from the liver tissue of RD and HF mice and the expression profiles were detected using RNA-seq. Differentially expressed genes between RD and HF mice were analyzed using BRB-ArrayTools. DAVID online tools were applied to analyze the GO and KEGG pathways. Transmission electron microscopy and western blotting were performed to observe the mitochondrial ultrastructure and quantified the expression of function-related proteins. Results Compared with the RD mice,the body weight gain was faster in the HF mice. The size of the lipid droplets was bigger in the HF-fed mouse liver tissue. Multiple pathway analysis all identified that these major gene changes were related to mitochondria. The mitochondrial deformation,enlarged or even destruction was observed in the high fat diet group observed by transmission electron microscopy. This observation was further confirmed by detecting of the expression of genes in the HF liver mitochondria. The levels of MFN1 and PHB1 were significantly increased, while the level of FKBP51 was significantly decreased. Conclusions FKBP51 is involved in the high-fat-induced mitochondrial damage via morphological and structural damages of mitochondria.

Detection and analysis of hematological and serum biochemical indices of thirty common marmosets / 中国比较医学杂志

Objective To examine hematological and biochemical parameters in 30 common marmosets,count the mean and standard deviations of each index, and analyze significant differences between female and male groups. Additionally,data from marmosets and macaque breeding were compared. Methods Blood was collected through the posterior limb vein while animals were awake. Hematology and serum biochemical indices were then measured with an automatic blood cell analyzer and blood biochemical analyzer, followed by statistical testing. Results No significant differences were measured in hematological indices between male and female groups. There was a significant difference between the female and male group in serum biochemical indices including high-density lipoprotein-cholesterol(HDL-C) and low-density lipoprotein-cholesterol(LDL-C)(P < 0.05). Compared with the foreign marmoset group, HGB, MCHC,NEUT,ALT, AST, and GLOB were visibly increased in the group of marmosets fed by our institution, but in accordance with the data range in rhesus monkeys. Conclusions Hematological and serum biochemical indices of common marmosets have been detected in this study and compared with related data in macaques and marmosets. Our findings provide basic data not only for pharmacological and toxicological studies,but also diagnosis and treatment of diseases.

Microsatellite marker design and genetic analysis for the common marmoset / 中国比较医学杂志

Objective To screen and optimize the microsatellite DNA primers of the laboratory common marmoset, analyze and evaluate the population genetic quality for the marmosets (Callithrix jacchus) introduced into the Institute of Medical Laboratory Animal science, Chinese Academy of Medical Sciences. Methods A total of 30 marmosets were randomly chosen, and their genome DNA from blood was extracted using phenol/chloroform method. The microsatellite DNA was amplified using standard polymerase chain reaction (PCR). The amplification products were tested by STR scanning after 2% agrose gel and 8% PAGE electrophoresis. The data processing and genetic analysis were completed using the Popgene1. 32 software. Results A total of 20 pairs of microsatellite loci showed genetic polymorphism, and 147 alleles were detected. The number of allele was 5 to 10, average 7. 35. The effective allele was 2. 2500 to 6. 3830, average 4. 0402. The observed heterozygosity was 0. 000 to 0. 4667, average 0. 1533. The expected heterozygosity was 0. 1424 to 0. 4350, average 0. 2506. The Shannon diversity index was 1. 2242 to 2. 0324, average 1. 5949. The polymorphic information content was 0. 5366 to 0. 8254, average 0. 7053. Conclusions The 20 pairs of marmoset microsatellite primers are genetically highly diverse and are in a Hardy-Weinberg equilibrium.

Preparation and identification of rabbit anti-marmoset IgG-HRP conjugate antibody / 中国比较医学杂志

Objective To purify marmoset serum IgG, prepare and identify the antiserum and the rabbit anti-marmoset antibody IgG-HRP (horseradish peroxidase). Methods Using SDS-PAGE analysis to identify the serum IgG from HiTrapTM Protein G. The antiserum titer was determined by double immunodiffusion assay. The rabbit anti-marmoset IgG was labeled with HRP by improved sodium periodate method. ELISA and western blotting were used to evaluate the concentration and specificity of rabbit anti-marmoset IgG-HRP. Results The purity of purified marmoset serum IgG determined by SDS-PAGE was higher than 95% , and the anti-serum titer of the anti-marmoset IgG polyclonal antibody was 1∶64. The concentration of rabbit anti-marmoset IgG-HRP identified by direct ELISA was 1∶256 000, and that by western-blotting was 1∶15 000, with a strong specificity. Conclusions The IgG-HRP marker antibody is prepared and the specificity and concentration are identified by ELISA and western blotting. It reserves the resources for the detection system of marmoset pathogens and the molecular immunological testing system.

Validation of the silencing site of marmoset B2m gene at the cellular level / 中国比较医学杂志

Objective To screen and determine the effective silencing targets of β2-microglobulin(B2m)gene at the cellular level in marmoset.Methods By homology comparison of the b2m gene in human and the B2m gene in marmoset, choose homology small hairpin RNA(shRNA)sequences targeting marmoset B2m gene were designed, We choose homology small hairpin RNA(shRNA)sequences targeting designed B2m gene to make homology analysis, and insert into lentivirus-based gene silencing constructs FUGW-TDT.The vectors were transfected into HEK293T cells induced by polyethylenimine(PEI).The suppression of B2m mRNA was detected by real-time PCR.Results Two gene-silencing sequences were screened that lied in 290~310 bp and 665~685 bp of the marmoset B2m mRNA, and have statistical significance in the silencing rate:(46.54±7.91)% (P < 0.05) and(83.22±4.37)%(P < 0.0001).Conclusions Two effective silencing target sequences are screened at cellular level, which can be further used in studies on gene silencing in marmoset.

Parenting violence modulates mediating effect of violent attitude between trait anger and aggressive behavior / 中华行为医学与脑科学杂志

Objective To explore the relationships among college students' trait anger,violent attitude and aggressive behavior.Methods A total of 991 college students were surveyed with the Trait Anger Scale,the Violent Attitude Questionnaire and the Aggression Questionnaire.Results (1)College students' scores on trait anger,violent attitude and aggressive scales were 17.99±3.94,3.52±2.20 and 70.12±14.67,respectively.(2) There were significant correlations among college students' trait anger,violent attitude and aggressive behavior(r=0.172-0.835,P<0.01).(3) Trait anger had a direct impact on aggressive behavior,as well as indirect effects through violent attitude.The direct effect of trait anger on aggressive behavior was 0.53,the indirect effect of violent attitude was 0.13,the total effect was 0.66,and the indirect effect was 20%.(4)The experience of parenting violence moderated the mediating effect of voilent attitude,and violent attitude moderated trait anger and aggression in those who didn't experience parenting violence(β=0.28,P<0.01),while the mediating effect was not significant in those who experienced parenting violence(β=-0.24,P=0.23,SIE=-0.166,P=0.28).Conclusion Violent attitude partially mediates trait anger and aggressive behavior,which is moderated by experienced parenting violence.

Analysis and comparison of RNA expression profiles in the heart and liver of Fkbp51 knockout mice / 中国比较医学杂志

Objective To study the function of Fkbp51 in the heart and liver by analyzing the differential RNA expression profiles in the wild-type mice (WT) and Fkbp51 knockout (KO) mice, and to elucidate the role of Fkbp51 gene in metabolic pathways in the heart and liver.Methods Using the second generation of high-throughput gene sequencing technology, the mRNA expression profiles of heart and liver were sequenced in WT and Fkbp51 KO mice.The data of sequencing of heart tissues were analyzed by DEGseq, and the results of sequencing of liver tissues were analyzed by BRB-Array Tools.The differential genes of the heart and liver in the mice were screened respectively.Gene ontology (GO) analysis and KEGG pathway analysis were performed to analyze the differentially expressed genes using the online tool DAVID.In addition, the differential genes of the two organ tissues were analyzed by Venn diagram.The interaction network of proteins was analyzed using the STRING database.Results (1) The absence of Fkbp51 led to changes in mRNA expressions of heart-related signal pathways such as vascular smooth muscle contraction, chemokine, retinol, and MAPK signaling pathways.(2) The lack of Fkbp51 mostly induced changes in cholesterol synthesis and metabolism, lipid metabolism, redox and other related genes and pathways in the liver.(3) In the heart and liver, Fkbp51 deletionresult ed in four co-differential genes, among them, down-regulation of Rnaset2b, Hmga1 and Fkbp51, while Cyp2b10 was down-regulated in the heart but up-regulated in the liver.All these proteins may interact with HSP90 protein and participat in the metabolism of heart and liver tissues.Conclusions Fkbp51 is involved in different metabolic and gene expression regulation pathways of heart and liver, and the roles are both independent and interrelated.

Adeno-associated virus mediated p53 gene silence in marmosets / 中国比较医学杂志

Objective To decrease the p53 gene expression at cellular and animal levels in marmoset using RNA interference technique.Methods The shRNA interference sequences were designed and inserted into the adeno-associated virus vector plasmid after bioinformatics analysis.The plasmids were transfected into African green monkey kidney cos-7 cells.The suppression of p53 mRNA was detected by real-time PCR, and the changes of p53 protein expression were detected by Western bolt.The adeno-associated virus-8 was injected through the hind leg vein.The changes of p53 protein expression in the liver tissue was detected by Western blot and immunohistochemistry.Results We screened two RNA interference effective arget sequences.The expression of p53 mRNA was suppressed ( 82.7 ±8.1 )% and ( 80.7 ± 7.5)%, respectively (P<0.05), and the expression of p53 protein was decreased (77.3 ±11.5)% and (73.7 ± 10.7)%, respectively (P<0.05).The two marmosets after virus infection showed that there were virus distributions in the liver, testes, and neck detected by in vivo fluorescence imaging.The expression of p53 in the marmoset liver was detected by western blot, immunohistochemistry analysis showing no obvious changes.Conclusions In the present study, the decrease of P53 gene expression at cellular level is achieved, however, the liver P53 protein in the marmoset liver is not significantly changes.Further optimization of the way of infection is needed in the future.

Bushen Huoxue Lishi Category TCM Compound in the Treatment of Chronic Prostatitis:A Systematic Re-view / 中国药房

OBJECTIVE:To systematically review the efficacy and safety of the Bushen huoxue lishi category TCM compound in the treatment of chronic prostatitis,and provide evidence-based reference for clinical treatment. METHODS:Retrieved from VIP Database,Wanfang Database,CJFD and CBM,randomized controlled trials(RCT)about Bushen huoxue lishi category TCM com-pound preparation (test group) versus conventional Western medicine (control group) in the treatment of chronic prostatitis were collected. Meta-analysis was performed by Rev Man 5.3 software after data extraction and quality evaluation. RESULTS:Totally 22 RCTs were enrolled,involving 1 863 patients. Results of Meta-analysis showed the total effective rate [OR=4.46,95%CI(3.40, 5.84),P<0.001],total scores of chronic prostatitis symptoms[MD=-3.62,95%CI(-5.21,-2.04),P<0.001] and lecithin count [MD=7.58,95%CI(2.15,13.01),P=0.006] in test group were significantly higher than control group,prostatic fluid white blood cell count [MD=-1.68,95%CI(-3.26,-0.10),P=0.04] was significantly lower than control group,with significant differenc-es. CONCLUSIONS:Bushen huoxue lishi category TCM compound has good efficacy in the treatment of chronic prostatitis.

Validity and reliability of the Chinese Attention to Positive and Negative Inventory in college students / 中国心理卫生杂志

Objective:To translate the Attention to Positive and Negative Inventory(APNI)and analyze the validity and reliability in Chinese undergraduates sample,to offer a convenient and reliable tool of measuring the cognitive bias for national researchers. Methods:The English-version APNI went through translation into Chinese, retroversion into English,translation into Chinese again,and revision several stages. Two parts of samples (1450 Chinese college students)were surveyed. Sample one (n=1000)was used for item analysis,exploratory factor a-nalysis (EFA),concurrent validity and reliability analysis,while sample 2 (n=450)was used for confirmatory fac-tor analysis (CFA). Totally 68 subjects of sample 1 were randomly chosen and resurveyed with an interval of one week. Beck depression inventory (BDI-II)and patient health questionnaire (PHQ-9)was used for concurrent validi-ty. Results:Item analysis indicated that the 22 items of Chinese APNI had good discriminability. EFA focused onattention to positive information(API)and attention to negative information(ANI)two factors. CFA showed good model fit (χ2 =1376,RMESA=0. 09,CFI=0. 94). Concurrent validity result showed that the total scores of BDI-II and PHQ-9 was negatively correlated with total scores of API (r=-0. 24,-0. 29,Ps<0. 01 ),and posi-tively correlated with total scores of ANI (r=0. 36,0. 31,Ps<0. 01). The Cronbach'αcoefficients of API and ANI sub-scale were 0. 86 and 0. 82,while the retest reliability coefficients were 0. 79 and 0. 62. Conclusion:It suggests that the Chinese APNI has good validity and reliability in a sample of college students,which could be used to eval-uate the cognitive bias of Chinese college students.

Validation of the silencing site of rhesus monkey P21 gene in COS-7 cells / 中国实验动物学报

Objective To screen the effective silencing targets of P21 gene at the cellular level in rhesus monkey . Methods To detect the expression of P21 gene in COS-7 cells ( derived from the kidney of African green monkey , Cerco-pithecus aethiops).Four small hairpin RNA (shRNA) sequences targeting rhesus monkey P21 gene were designed and in-serted into lentivirus-based gene silencing constructs FUGW-TDT.The vectors were transfected into COS-7 cells respective-ly.The suppression of P21 mRNA was detected by real-time PCR, and the expression of P21 protein was detected by West-ern blot assay .Results Four gene-silencing sequences were screened that lied in 541-561 bp, 542-562 bp, 215-239 bp, and 624-648 bp of the rhesus monkey P21 mRNA.Their silencing rate was (91.82 ±3.21)%, (82.47 ±2.48)%, (81.31 ±2.69 )% and ( 87.35 ±4.59 )%, and the protein expression was ( 11.97 ±0.70 )%, ( 20.22 ±0.65 )%, ( 23.21 ± 0.63)%and (14.42 ±0.86)%, respectively.Conclusions Four effective silencing target sequences are screened at cel-lular level , which can be used in gene silencing research of rhesus monkeys .

The mediator effect of coping strategy between stressful life events and negative emotions in young military men / 中华行为医学与脑科学杂志

Objective To investigate mediate role of coping strategy between stressful life events and negative emotions which anxiety and depression are included.Methods Questionnaires of psychosocial stress survey for groups (PSSG),Simple Coping Style Questionnarire(SCSQ),Selfrating Anxiety Scale (SAS) and Selfrating Depression Scale(SDS) were used to examine 1 764 young servicemen.Results Correlation analysis showed that there existed negative correlations between positive coping and stressful life events,passive coping,anxiety,depression(r=-0.080～-0.304,P＜0.05).And correlation analysis also showed there existed positive correlations among the other variables(r=0.230 ～ 0.756,P＜0.05).There existed partly mediating effect of positive coping,passive coping between stressful life events and negative emotions.The standardized total effect of life events to anxiety was 0.302.Mediating effects of positive coping and passive coping were 0.033,0.044.The standardized total effect of life events to depression was 0.230,mediating effects of positive coping and passive coping were 0.051,0.029.Conclusion Coping strategy is an important mediator between stress and negative emotions such as anxiety and depression.

Establishment of an indirect immunofluorescence assay for detection of murine norovirus / 中国比较医学杂志

Objective To establish an indirect immunofluorescence assay for detection of murine norovirus ( MNV) .Methods Mouse leukaemic monocyte macrophage cell line RAW 264.7 cells were infected with MNV-1 and cultured for 36 hours to collect the virus and uninfected cells , and to make antigen glass slides .BALB/c mice were gavaged with MNV-1 (107 TCID50) and infected sera were collected as positive control .The serum was 1:10 diluted and used for measuring MNV antibody by immunofluorescence assay ( IFA ) .80 serum samples were tested using the two methods , IFA and ELISA, and the discrepant samples were validated by Western blotting .Results RAW264.7 cells were infected with MNV-1 for 36-48 h, showing an infection rate of 60% of the cells, and the cells infected for 36 h were preferred.IFA method was used to detect the serum with MNV-1 infection and showed that the antibody content was gradually increased at one week after infection , reaching a maximum antibody concentration at 4 weeks after infection , and maintained a stable level later .The mouse serum at four weeks after MNV-1infection was used as positive quality control . Among the 80 serum samples , 27 positive and 53 negative cases were detected by IFA method , and 32 positive and 48 negative cases were detected by ELISA .The five discrepant samples were verified by Western blotting , resulted in 3 positive and 2 negative cases . The coincidence rate of IFA was 96.0% and that of ELISA methods was 97.5%. Conclusions Basically, immunofluorescence assay can be used to detect the MNV-1 infection in mice, although false negative result may occur occasionally .IFA and ELISA detection can be selected as initial screening measures , and use Western blot assay to verify the discrepant samples .

Animal biosafety isolation device and its evaluation / 中国比较医学杂志

This paper discusses the definition , classification, selection, monitoring and evaluation of animal biosafety isolation device .Evaluation order of animal biosafety isolation device follows animal survival needs -biosafety needs-animal welfare requirements .

Rhesus monkey P53 gene silencing at the cellular level / 中国比较医学杂志

Objective In order to establish a rhesus monkey model of p53 gene silencing, firstly we screened and determined the effective silencing targets of p53 gene at the cellular level in rhesus monkey.Methods The expression of p53 gene was detected in COS-7 cells ( derived from the kidney of the African Green Monkey, Cercopithecus aethiops).Three small hairpin RNA ( shRNA) sequences targeting rhesus monkey p53 gene were designed, analysed by bioinformatics, and inserted into lentivirus-based gene silencing constructs FUGW-TDT.The plasmids of p53-RNAi and control vector were transfected into the COS-7 cells, respectively.The suppression of p53 mRNA was detected by real-time PCR, and the changes of p53 protein expression were detected by Western blot assay.Results p53 gene expression was detected in COS-7 cells.Bioinformatics analysis showed that three gene-silencing sequences were screened which lied in the open reading frame ( ORF) region and targeted 238 -258bp, 681 -701bp, 169 -189bp of the rhesus monkey p53 mRNA.At 48 hrs after transfection of the three silencing constructs, p53 mRNA was suppressed by(87.17 ±4.03)%, ( 72.62 ±4.11)% and(76.22 ±0.98 )%, and p53 protein was suppressed by ( 84.44 ±2.18 )%, ( 71.04 ±1.18)% and ( 74.17 ±0.95 )%, respectively. Conclusions We obtained three effective target sequences showing high efficiency in p53silencing, which can be used in further studies on gene silencing in rhesus monkey.

Application of Chinchilla in medical research / 中国实验动物学报

Chinchilla has been successfully used as an animal model in the study of auditory system, microorgan-ism and parasitic infection, because of its unique biological features, and it can be further developed for the research of se-nile diseases, metabolic diseases, etc.This paper will introduce the related biological characteristics of chinchilla, and briefly reviewed the progress of its application in medical research.

A comparative study of the effects of group psychological quality training between field soldiers and military students cohorts / 中华行为医学与脑科学杂志

Objective To explore the effects of group psychological training on mental health of military students and field soldiers.Methods A total of 60 students and 48 soldiers received group psychological quality training and studied a textbook Mental Quality Training for armymen for 3 months.Mental Quality Questionnaire for Amymen (MQQA),Symptom CheckList 90 (SCL-90),Self-rating Anxiety Scale (SAS),Self-rating Depression Scale (SDS) and State-Trait Anxiety Inventory(STAI) were employed to evaluate mental health and psychological quality of subject before and after the training.All the data were analyzed by independent-samples t test.Results (1) The difference in MQQA score of field soldiers before and after training was significantly larger than that of military students in loyalty and general score,and lower in willpower((-16.58 ± 7.75) vs.(-1.75 ± 8.68),(-27.74 ± 28.74) vs.(-12.57 ± 30.96),P ＜ 0.05).(2) The SCL-90 difference of field soldiers between before and after training was significantly larger than that of military students in hostility and phobic anxiety((0.26 ±0.47) vs.(0.07 ± 0.24),(0.13 ± 0.40) vs.(0.02 ± 0.13),P ＜ 0.05).(3) The difference in emotion score of field soldiers between before and after training was significantly larger than that of military students in SAS,SDS,SAI,TAI and STAI (P ＜ 0.01).Conclusion The effects of group psychological quality training on field soldier group are better than that of military students,which is helpful to improve mental quality and mental health,as well as to relax anxiety and depression of soldiers.