Effect of FKBP38 protein on endometrial precancerous lesions and mechanism / 中国药理学通报

Aim To build the model of the gene FKBP38(FK506 binding protein 38)conditional knock out in uterus and then investigate the effect on endometrial precancerous lesions and the underlying mechanism.Methods Transgenic mice whose FKBP38 gene was flanked with loxP were constructed by embryo microinjection. The conditional knockout of FKBP38 was obtained by breeding mice harboring two loxP sites in FKBP38(FKBP38

Cathepsin D overexpression in the nervous system rescues lethality and Aβ42 accumulation of cathepsin D systemic knockout in vivo

The lysosome is responsible for protein and organelle degradation and homeostasis and the cathepsins play a key role in maintaining protein quality control. Cathepsin D (CTSD), is one such lysosomal protease, which when deficient in humans lead to neurolipofuscinosis (NCL) and is important in removing toxic protein aggregates. Prior studies demonstrated that CTSD germ-line knockout-CtsdKO (CDKO) resulted in accumulation of protein aggregates, decreased proteasomal activities, and postnatal lethality on Day 26 ± 1. Overexpression of wildtype CTSD, but not cathepsin B, L or mutant CTSD, decreased α-synuclein toxicity in worms and mammalian cells. In this study we generated a mouse line expressing human CTSD with a floxed STOP cassette between the ubiquitous CAG promoter and the cDNA. After crossing with Nestin-cre, the STOP cassette is deleted in NESTIN + cells to allow CTSD overexpression-CTSDtg (CDtg). The CDtg mice exhibited normal behavior and similar sensitivity to sub-chronic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced neurodegeneration. By breeding CDtg mice with CDKO mice, we found that over-expression of CTSD extended the lifespan of the CDKO mice, partially rescued proteasomal deficits and the accumulation of Aβ42 in the CDKO. This new transgenic mouse provides supports for the key role of CTSD in protecting against proteotoxicity and offers a new model to study the role of CTSD enhancement in vivo.

Role of zinc finger protein 36, C3H type-like 1 mediating astrocytes activation in motor neuron degeneration in amyotrophic lateral sclerosis / 解剖学报

Objective To investigate the role of zinc finger protein 36,C3H type-like 1 (ZFP36L1) mediating astrocytes activation in the degeneration of motor neurons in amyotrophic lateral sclerosis (ALS). Methods Superoxide dismutase 1 (S0D1)-G93A transgenic mice were used as animal models, the wild-type littermates as the control (13 mice were taken from mutant and wild-type mice at each time point) . The ZFP36L1 mRNA and protein levels of the spinal cord in the early, middle and late stage were detected by Real-time PGR and Western blotting. The expression and distribution of ZFP36L1 in the spinal cord were detected by immunofluorescence. Primary astrocyte model was established from 15 postnatal 1-2 day mice. The ZFP36L1 mRNA and protein levels in astrocytes were detected by Real-time PCR and Western blotting. Si-ZFP36L1 was transfected into SOD1-G93A mutant primary astrocytes. The transfection efficiency was detected by Western blotting. Tumor necrosis factor a (TNF-a) and interleukin-18 (IL-18) secreted from astrocytes after transfection were assessed by Western blotting and ELISA. After silencing ZFP36L1 in SOD1-G93A mutant primary astrocytes, it was cocultured with SOD1-G93A mutant NSC34 cells. 5 ' -ethynyl-2' deoxyuridine (EdU) test and the level of proliferating cell nuclear antigen (PCNA) were used to determine the effect of ZFP36L1 on NSC34 cell proliferation. TUNEL test and the level of cleaved-Caspase-3 were used to determine the effect of ZFP36L1 on NSC34 cell apoptosis. Blank small interfering RNA(siRNA) was transfected as the control group. Results Compared with the wild-type mice, the mRNA and protein levels of ZFP36L1 were downregulated in the spinal cord of SOD1-G93A transgenic mice. In wild type mice, ZFP36L1 positive cells were mainly [

Expression of protein tyrosine kinase 7 and receptor tyrosine kinase-like orphan receptor 2 in the brainstem of amyotrophic lateral sclerosis transgenic mice with hSODl-G93A mutation / 解剖学报

Objective To investigate the relationship between changes in protein tyrosine kinase 7 (PTK7) and receptor tyrosine kinase-like orphan receptor 2 ( ROR2) expression in the brainstem and the pathogenesis of amyotrophic lateral sclerosis (ALS). Methods Forty-four human superoxide dismutase 1( hSODl)-G93A mutant ALS transgenic mice were selected, and an equal number of wild-type littermates was used as control. The brainstems were isolated at da)' 70, day 95, day 108 and da)' 122 after birth, and the morphology of frypoglossal nucleus (12N) and nucleus of facial nerve(7N) neurons in the brainstem of the model mice were observed by Nissl staining. The mRNA and protein expression of PTK7 and ROR2 were detected by RT-PCR and Western blotting respectively, and the cellular localization and distribution of PTK7 and ROR2 in 12N and 7N were observed by immunofluorescence double-labeling technique. Results The result of Nissl staining showed that Nissl bodies in the neurons reduced distinctly with vacuolar degeneration of neurons, cell body atrophy and nuclear volume reduction in the 12N and 7N brainstems of ALS transgenic mice. RT-PCR result indicated that ROR2 and PTK7 mRNA level in the brainstem of ALS transgenic mice were up-regulated at da)' 70, da)' 95, day 108 and day 122 compared with wild-type littermates. Western blotting result showed that PTK7 protein was up-regulated at day 70, day 95, day 108 and day 122, ROR2 protein was up-regulated at day 70, day 95, day 108, and down-regulated at day 122 in the brainstem of ALS transgenic mice compared with wild-type littermates. Immunofluorescence result showed that ROR2/neuronal nuclei (NeuN)and PTK7/NeuN double positive cells, ROR2/glial fibrillary acidic protein (GFAP) and PTK7/GFAP double positive cells were observed in the 12N and 7N of the brainstem of ALS transgenic mice and wild-type mice, suggesting that ROR2 and PTK7 were expressed both in neurons and astrocytes. Conclusion PTK7 and ROR2 are abnormally expressed in the brainstem of ALS transgenic mice, which is closely related to the pathogenesis of ALS.

Establishment and identification of secreted frizzled-related protein 4 transgenic mice and analysis of their biological properties / 西安交通大学学报(医学版)

【Objective】 To generate secreted frizzled-related protein 4 (SFRP4) transgenic mice and analyze their biological properties. 【Methods】 We generated SFRP4 transgenic mice by DNA microinjection and detected the expression of SFRP4 by qRT-PCR and Western blotting. 【Results】 SFRP4 transgenic mice were successfully generated by DNA microinjection. The expression of SFRP4 was dramatically increased in transgenic mice liver compared with that of wide type. 【Conclusion】 The transgenic mice model of SFRP4 was successfully established by DNA microinjection. It provides a good model for studying the function of SFRP4 and the mechanism of participating in metabolic diseases.

Relationship between DEAD-box helicase 5, transcription factor 12 and amyotrophic lateral sclerosis / 解剖学报

Objective To explore the relationship between the expression of DEAO-box helicase 5(DDX5) and transcription factor 12(TCF12) with amyotrophic lateral sclerosis ( ALS ) hippocampal lesions by detecting the expressions and the interaction of DDX5 and TCF12 in the hippocampus of SOD1-G93A mutant ALS transgenic mice. Methods Forty- two pairs of SOD1-G93A mutant ALS transgenic mice and wild-type mice were divided into three groups at the age of 95 days (early onset stage), 108 days (middle onset stage) and 122 days (late onset stage). RT-PCR, Western blotting and double immunofluorescence labeled technique were used to detect the expressions of DDX5 and TCF12 in the hippocampus. Co-immunoprecipitation assasy was used to detect the interaction between DDX5 and TCF12. Results Compared with the wild-type mice of the same age, DDX5 and TCF12 mRNA in the hippocampus of SOD1-G93A mutant ALS transgenic mice were unchanged, but DDX5 and TCF12 protein were up-regulated significantly at day 95, 108 and 122. DDX5 and TCF12 positive cells were found in both DG area and hippocampus proper, and DDX5 and TCF12 were co-localized with neurons. The immunoreactivities of DDX5 and TCF12 in the hippocampus of SOD1-G93A mutant transgenic mice were elevated compared with wild-type mice at the same time point. Co-immunoprecipitation assasys confirmed that there existed interactions between DDX5 and TCF12 protein. Conclusion DDX5 and TCF12 protein are up-regulated in the hippocampal tissues of SOD1-G93A mutant ALS transgenic mice. The abnormal expressions of DDX5 and TCF12 are involved in the hippocampal lesions of ALS.

The role of tyrosine phosphatase Shp2 in spermatogonial differentiation and spermatocyte meiosis / 亚洲男科学杂志(英文版)

The transition from spermatogonia to spermatocytes and the initiation of meiosis are key steps in spermatogenesis and are precisely regulated by a plethora of proteins. However, the underlying molecular mechanism remains largely unknown. Here, we report that Src homology domain tyrosine phosphatase 2 (Shp2; encoded by the protein tyrosine phosphatase, nonreceptor type 11 [Ptpn11] gene) is abundant in spermatogonia but markedly decreases in meiotic spermatocytes. Conditional knockout of Shp2 in spermatogonia in mice using stimulated by retinoic acid gene 8 (Stra8)-cre enhanced spermatogonial differentiation and disturbed the meiotic process. Depletion of Shp2 in spermatogonia caused many meiotic spermatocytes to die; moreover, the surviving spermatocytes reached the leptotene stage early at postnatal day 9 (PN9) and the pachytene stage at PN11-13. In preleptotene spermatocytes, Shp2 deletion disrupted the expression of meiotic genes, such as disrupted meiotic cDNA 1 (Dmc1), DNA repair recombinase rad51 (Rad51), and structural maintenance of chromosome 3 (Smc3), and these deficiencies interrupted spermatocyte meiosis. In GC-1 cells cultured in vitro, Shp2 knockdown suppressed the retinoic acid (RA)-induced phosphorylation of extracellular-regulated protein kinase (Erk) and protein kinase B (Akt/PKB) and the expression of target genes such as synaptonemal complex protein 3 (Sycp3) and Dmc1. Together, these data suggest that Shp2 plays a crucial role in spermatogenesis by governing the transition from spermatogonia to spermatocytes and by mediating meiotic progression through regulating gene transcription, thus providing a potential treatment target for male infertility.

Role of HIV-1 envelope protein gp120 in neuronal injury-induced cognitive impairment / 中华微生物学和免疫学杂志

Objective@#To investigate the role of HIV-1 envelope protein gp120 in cognitive impairment induced by neuronal damage.@*Methods@#Western blot and immunofluorescence assay were used to detect microglia activation, inflammatory factor expression and neuronal damage after gp120 treatment. Neuronal damage and neurocognitive performance in gp120-transgenic mice were evaluated using immunohistochemical staining and behavioral analysis, respectively.@*Results@#In vivo and in vitro experiments showed that HIV-1 gp120 significantly induced the expression of caspase-1 and IL-1β, and indirectly caused neuronal synaptic shortening and neuronal damage (P<0.05). Compared with wild-type mice, gp120-transgenic mice showed significant cortical and hippocampal glial activation, neuronal loss, dendritic damage and neurocognitive disorders.@*Conclusions@#HIV-1 gp120 might cause neuronal damage through activating the release of inflammatory factor by microglia and involve in neurocognitive impairment.

Effect of different test indicators and observation periods in contextual fear conditioning test on detection of behavioral efficacy of 5XFAD transgenic mice / 上海交通大学学报（医学版）

Objective • To explore whether different test indicators and observation periods in contextual fear conditioning test affect the detection effectiveness of learning and memory ability of 5XFAD transgenic mice. Methods • Twelve 4-month-old female 5XFAD transgenic progeny mice and 14 4-month-old female LM progeny mice were selected, which were produced by crossing male 5XFAD transgenic mice and female C57BL/6 mice, to conduct open field test and contextual fear conditioning test in succession. Total distance and velocity in open field test and average motion index in the first 60 s of training stage in contextual fear conditioning test of the two groups of mice were used to evaluate the difference of locomotor activity. Besides, the first 180 s, 181-360 s and the first 300 s of testing stage for observation were selected to evaluate the selection effects on percent freeze and activity suppression ratio. Then further investigation was launched to explore the effects of different observation periods on the detection effectiveness of percent freeze and activity suppression ratio. Results • The differences of total distance and total velocity of the two groups of mice in open field test were not statistically significant, however the average motion index in the first 60 s of training stage in contextual fear conditioning test of 5XFAD transgenic mice was significantly higher than that of LM mice (P=0.027). The comparison of percent freeze among the three groups of observation periods of LM mice had significant difference (both P<0.05), while there was no statistical significance in activity suppression ratio. The comparisons of percent freeze and activity suppression ratio among the three groups of observation periods of 5XFAD mice had no significant difference. The differences of percent freeze between 5XFAD mice and LM mice was not statistically significant during the three observation periods. However, the activity suppression ratio of 5XFAD mice was significantly higher than that of LM mice in the first 180 s (P=0.038), in the other two observation periods the difference of activity suppression ratio between the two groups of mice was not statistically significant. Conclusion • The average motion index detected in training stage in contextual fear conditioning test is more sensitive than total distance and total velocity detected in open field test for evaluating locomotor activity of 5XFAD mice. In contextual fear conditioning test, different observation periods have effects on the value of percent freeze provided by single strain of mice, while the value of activity suppression ratio remains unaffected. Activity suppression ratio is more precise than percent freeze to reflect the cognitive deficiency of 5XFAD mice. It is more accurate to select the first 180 s of testing stage for observation.

Establishment of a gp120 transgenic mouse model with 7 nAChR knockout / 南方医科大学学报

OBJECTIVE@#To construct a HIV-1 gp120 transgenic mouse model (gp120) with 7 nicotinic acetylcholine receptor (7nAChR) gene knockout.@*METHODS@#The 7nAChR gene knockout mice (7R) were crossed with HIV-1gp120 transgenic mice (gp120) to generate F1 generation mice. We selected the F1 mice with the genotype of 7R/gp120 to mate to obtain the F2 mice. The genotypes of the F3 mice were identified by PCR, and the protein expressions in the double transgenic animal model was analyzed by immunohistochemistry. BV2 cells were treated with gp120 protein and 7nAChR inhibitor, and the expressions of IL-1β and TNF- were detected using ELISA.@*RESULTS@#The results of PCR showed the bands of the expected size in F3 mice. Two F3 mice with successful double gene editing (7R/gp120) were obtained, and immunohistochemistry showed that the brain tissue of the mice did not express 7 nAChR but with high gp120 protein expression. In the cell experiment, treatment with gp120 promoted the secretion of IL-1β and TNF- in BV2 cells, while inhibition of 7nAChR significantly decreased the expression of IL-1β and TNF- ( < 0.001).@*CONCLUSIONS@#By mating gp120 Tg mice with 7R mice, we obtained gp120 transgenic mice with 7nAChR gene deletion, which serve as a new animal model for exploring the role of 7nAChR in gp120-induced neurotoxicity.

The role of tyrosine phosphatase Shp2 in spermatogonial differentiation and spermatocyte meiosis / 亚洲男科学杂志(英文版)

The transition from spermatogonia to spermatocytes and the initiation of meiosis are key steps in spermatogenesis and are precisely regulated by a plethora of proteins. However, the underlying molecular mechanism remains largely unknown. Here, we report that Src homology domain tyrosine phosphatase 2 (Shp2; encoded by the protein tyrosine phosphatase, nonreceptor type 11 [Ptpn11] gene) is abundant in spermatogonia but markedly decreases in meiotic spermatocytes. Conditional knockout of Shp2 in spermatogonia in mice using stimulated by retinoic acid gene 8 (Stra8)-cre enhanced spermatogonial differentiation and disturbed the meiotic process. Depletion of Shp2 in spermatogonia caused many meiotic spermatocytes to die; moreover, the surviving spermatocytes reached the leptotene stage early at postnatal day 9 (PN9) and the pachytene stage at PN11-13. In preleptotene spermatocytes, Shp2 deletion disrupted the expression of meiotic genes, such as disrupted meiotic cDNA 1 (Dmc1), DNA repair recombinase rad51 (Rad51), and structural maintenance of chromosome 3 (Smc3), and these deficiencies interrupted spermatocyte meiosis. In GC-1 cells cultured in vitro, Shp2 knockdown suppressed the retinoic acid (RA)-induced phosphorylation of extracellular-regulated protein kinase (Erk) and protein kinase B (Akt/PKB) and the expression of target genes such as synaptonemal complex protein 3 (Sycp3) and Dmc1. Together, these data suggest that Shp2 plays a crucial role in spermatogenesis by governing the transition from spermatogonia to spermatocytes and by mediating meiotic progression through regulating gene transcription, thus providing a potential treatment target for male infertility.

Identification and survival analysis of TTA/TETO-CCKR-2 double transgenic mice / 中国组织工程研究

BACKGROUND: The inducible forebrain-specific cholecystokinin receptor-2 (CCKR-2) double transgenic (tTA/tetO-CCKR-2 tg, abbreviated as dtg) mice are an ideal model of anxiety-related diseases. However, there is still a lack of model identification and life related data OBJECTIVE: To identify the genomic DNA of the offspring and the specific expression of CCKR-2 transgene in the forebrain, and to analyze the survival probability of dtg mice. METHODS: α-CaMKII/tTA single transgenic mice and tetO-CCKR-2 single transgenic mice were cross-fertilized to construct a dtg mouse model. The genomic DNA was extracted from the tail of the offspring, and the genotypes were detected by PCR and agarose gel electrophoresis. Wild-type (WT) mice were used as controls. In situ hybridization was used to detect the expression of CCKR-2. Survival of dtg mice and WT mice (30 females and 30 males) was observed and recorded within 2 years. The study protocol was approved by the Experimental Animal Ethics Committee of Southwest Medical University, with an approval No. 20150068. RESULTS AND CONCLUSION: Agarose gel electrophoresis results showed the molecular weight of the PCR products of dtg mice was consistent with the expected target gene fragment. In situ hybridization results showed a strong signal of CCKR-2 was detected in the forebrain of dtg mice, but hardly present in the WT mice. The median survival time of dtg mice was 76 weeks in females and 77 weeks in males. The survival probability was decreased with age in dtg mice. The survival probability of WT mice was significantly better than that of dtg mice (P < 0.001). There was no significant sex difference between males and females of dtg mice (P=0.577). Therefore, the specific expression of CCKR-2 transgene in the forebrain can be identified using PCR amplification, genomic DNA extraction, agarose gel electrophoresis, and in situ hybridization. tTA/tetO-CCKR-2 double transgenic induction may shorten the survival time of mice, but no significant difference is observed between the females and males of dtg mice.

Effect of epigallocatechin gallate on expressions of synaptic plasticity and neural cell adhesion molecules in APP / PS1 double transgenic mice / 解剖学报

Objective To observe the effect of epigallocatechin gallate (EGCG) on the spatial learning memory deficit in amyloid procursor protein (APP) / presenilin-1 (PS1) double transgenic mice, synaptic ultrastructure and expression of neural cell adhesion molecule in hippocampal CA1 region. Methods Eight weeks old male APP / PS1 double transgenic mice were selected as Alzheimer’s disease (AD) model and divided into the model group, the EGCG group and the donepezil hydrochloride group, 12 in each group.Besides,normal mice of the same brood (with no transgene) were recruited as a normal group (n = 12). Related indices were detected after 6 months continuous gastrogavage. The spatial learning-memory deficit of APP / PS1 double transgenic mice was detected by Morris water maze test. The synaptic ultrastructure of hippocampal CA1 region was observed by transmission electron microscopy. The expression levels of neural cell adhesion molecule (NCAM) and polysialyltranseferase α2,8-polysialic acid (ST8Sia Ⅱ) protein in hippocampal CA1 region of APP / PS1 transgenic mice were detected by immunofluorescence and Western blotting. Results Compared with the normal group, the mean value of escape latency in the model group was extended, and compared with the model group, the mean value of escape latency in the EGCG group and donepezil hydrochloride group were increased (P < 0. 05) . The result of electron microscope showed that the changes of synaptic interface curvature of EGCG group and donepezil hydrochloride group were not obvious. Compared with the model group, the width of the synaptic gap becomes narrower and the thickness of the post-synaptic compact were increases (P < 0. 05) . Immunofluorescence showed that the expression of NCAM and ST8Sia Ⅱ proteins in the hippocampus CA1 region was expressed in the cytoplasm of neurons, the expressions of NCAM and ST8Sia Ⅱ in hippocampal CA1 region were significantly increased in EGCG group and donepezil hydrochloride group (P< 0. 05) . Their contents also showed higher levels of expression in Western blotting (P < 0. 05) . Conclusion EGCG shows improvement on the spatial learning-memory deficit in APP / PS1 double transgenic mice,which may be associated with affecting the synaptic structure of hippocampus and improving the expressions of neural cell adhesion molecule.

Role of HIV-1 envelope protein gp120 in neuronal injury-induced cognitive impairment / 中华微生物学和免疫学杂志

Objective To investigate the role of HIV-1 envelope protein gp120 in cognitive im-pairment induced by neuronal damage. Methods Western blot and immunofluorescence assay were used to detect microglia activation, inflammatory factor expression and neuronal damage after gp120 treatment. Neu-ronal damage and neurocognitive performance in gp120-transgenic mice were evaluated using immunohisto-chemical staining and behavioral analysis, respectively. Results In vivo and in vitro experiments showed that HIV-1 gp120 significantly induced the expression of caspase-1 and IL-1β, and indirectly caused neuro-nal synaptic shortening and neuronal damage (P<0. 05). Compared with wild-type mice, gp120-transgenic mice showed significant cortical and hippocampal glial activation, neuronal loss, dendritic damage and neu-rocognitive disorders. Conclusions HIV-1 gp120 might cause neuronal damage through activating the re-lease of inflammatory factor by microglia and involve in neurocognitive impairment.

Pharmacodynamic study of magnesium isoglycyrrhizinate in HBV transgenic mouse model / 中国药科大学学报

@#This study aims to discuss the therapeutic effect of magnesium isoglycyrrhizinate on hepatitis B virus(HBV)transgenic mouse and its effect on cellular immunity and liver inflammation. The changes of serum aspartate aminotransferase(AST)and alanine aminotransferase(ALT)activity, the difference of serum hepatitis B surface antigen(HBsAg), liver tissue HBsAg mRNA, and the pathological morphological changes of liver tissue were detected to investigate the hepatic inflammatory lesions and the efficacy of magnesium isoglycyrrhizinate in HBV transgenic mouse. Peripheral blood lymphocytes were classified by flow cytometry, and serum cytokines were detected by cytometric bead array(CBA)to explore the mechanism of magnesium isoglycyrrhizinate to reduce hepatic inflammatory lesions in HBV transgenic mouse. After grouping HBV transgenic mouse with serum transaminase activity and 35 days of continuous administration, serum transaminases level in magnesium isoglycyrrhizinate ［15 mg/(kg ·d)］ group was significantly lower than that in control group(P< 0. 05), serum HBsAg protein and liver tissue HBsAg mRNA increased with time, but there was no significant difference between the two groups. The main pathological changes of liver were liver cell swelling, necrosis and focal inflammatory cell infiltration, and the pathological changes of liver in magnesium isoglycyrrhizinate group were lighter than those in control group. The number of CD8+ cells in the blood of magnesium isoglycyrrhizinate group was significantly less than that in the control group(P< 0. 05)and the CD4+/CD8+ cell ratio was significantly higher than that in the control group(P< 0. 05). The content of inflammatory cytokines in serum of magnesium isoglycyrrhizinate group decreased significantly(P< 0. 05). Magnesium isoglycyrrhizinate can regulate the immune function of HBV transgenic mouse, decrease the infiltration of inflammatory cells in hepatic tissue and hepatocyte injury, but do not affect the expression of hepatocyte HBsAg.

Establishment of transgenic mice-derived allografts in pancreatic cancer / 西安交通大学学报(医学版)

Objective: To establish the transgenic mice-derived allografts (MDAs) so as to provide an appropriate experiment model of pancreatic cancer. Methods: By using the tissue fragments derived from transgenic mice with pancreatic cancer named LSL-KrasG12D/+; Trp53fl/+; Pdx1-Cre (KPC) mice and subcutaneous transplant technique, we constructed the MDAs. We evaluated the histopathological characteristics of MDAs and infiltration of immune cells by H&E staining, Masson's staining and immunohistochemical method. Results: MDAs could mimic the pathological morphology, proliferation and fibrosis of pancreatic cancer precisely. Besides, the infiltration of immune cells in MDAs tumor was the same as that of pancreatic cancer. Conclusion: MDAs are an effective model to mimic the development of pancreatic cancer.

Liuwei Dihuang Decoction improves cognitive impairments via regulating immune system in APP/PS1 transgenic mouse,a mouse model of Alzheimer disease / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect and mechanisms of Liuwei Dihuang Decoction (LW)on cognition in PrP-hAβPPswe/PS1ΔE9(APP/PS1)transgenic mice.METHODS LW was adminis-trated with oral for 3 months.The locomotor activity test was performed to investigate the spontaneous motor activity of mice. The Morris water maze test and shuttle box test were performed to investigate the spatial learning and memory and active avoidance response respectively.The Αβ deposits and neuron loss in the hippocampus was detected by immunofluorescence staining and nissl staining respectively. The flow cytometry was employed to investigate the lymphocyte subsets of the mice.The 3H-thymidine incorporation was performed to investigate the splenocytes proliferation. RESULTS The treatment of LW ameliorated the impairments of spatial learning and memory and active and passive avoidance in APP/PS1 mice. The administration of LW alleviated neuron loss in the brain, suppressed amyloid-β (Αβ) deposits in the hippocampus of APP/PS1 mice. The treatment of LW significantly increased ConA-and LPS-induced proliferation of splenocytes,increased CD3+T cells and CD19+B cells in the spleen lymphocytes and reduced Gr1+cells in APP/PS1 mice.CONCLUSION This data indicated the adminis-tration of LW ameliorated behavioral and pathological deterioration via regulating immune function.

Elevation of Divalent Metal Transporter 1 Protein in the Cerebellar Cortex of the APP/PS1 Transgenic Mouse / 中国医科大学学报

Objective To investigate the distribution of divalent metal transporter 1 (DMT1) in the cerebellum of APP/PS1 transgenic mouse. Methods Immunohistochemistry, immunofluorescence, and confocal laser scanning microscopy were used to analyze the relationship between DMTl and amyloid beta (Aβ) and their distribution in senile plaques. Western blotting was used to analyze DMT1 protein level in the APP/PS1 transgenic mouse cerebellum. Results DMTl and Aβ were mainly located in the amyloid plaques, which were predominately located in the molecular layer of the cerebellar cortex of the transgenic mouse. Only a few plaques could be seen in the Purkinje cell layer and granular layer. Confocal laser microscopy revealed the DMTl and Aβ were co-localized in senile plaques. Conclusion The abundant expression of DMTl protein suggests that DMTl and the divalent metal ions that it transports might be involved in the formation of Aβ senile plaques and other pathological processes in the cerebellum in Alzheimer' s disease.

Gastric Carcinogenesis in the miR-222/221 Transgenic Mouse Model / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: MicroRNAs (miRNAs) regulate various cellular functions, including development, cell proliferation, apoptosis, and tumorigenesis. Different signatures associated with various tissue types, diagnosis, progression, prognosis, staging, and treatment response have been identified by miRNA expression profiling of human tumors. miRNAs function as oncogenes or as tumor suppressors. The relationship between gastric cancer and miRNA garnered attention due to the high incidence of gastric cancer in Asian countries. miR-222/221 expression increases in gastric tumor tissues. The oncogenic effect of miR-222/221 was previously determined in functional studies and xenograft models. In this study, transgenic mice over-expressing miR-222/221 were generated to confirm the effect of miR-222/221 on gastric carcinogenesis. MATERIALS AND METHODS: At 6 weeks of age, 65 transgenic mice and 53 wild-type mice were given drinking water containing N-nitroso-N-methylurea (MNU) for 5 alternating weeks to induce gastric cancer. The mice were euthanized at 36 weeks of age and histologic analysis was performed. RESULTS: Hyperplasia was observed in 3.77% of the wild-type mice and in 18.46% of the transgenic mice (p=0.020). Adenoma was observed in 20.75% of the wild-type mice and 26.15% of the transgenic mice (p=0.522). Carcinoma was observed in 32.08% of the wild-type mice and 41.54% of the transgenic mice (p=0.341). The frequency of hyperplasia, adenoma, and carcinoma was higher in transgenic mice, but the difference was statistically significant only in hyperplasia. CONCLUSION: These results suggest that hyperplasia, a gastric pre-cancerous lesion, is associated with miR-222/221 expression but miR-222/221 expression does not affect tumorigenesis itself.

Light exposure resulted in APPswe/PS1 transgenic mice retinal structural and functional disorder / 眼科新进展

Objective To research the functional and structural change in TgAP-PswePS1 transgenic mice after intensive light exposure insult.Methods APPswe/PS1 transgenic mice at the age of 6 months old were grouped for experiments,the transgenic mice were replaced by light insult device for 6 months,while the control mice were kept in normal conditions.After 6 months light exposure,the eyes of control and experimental mice were examined with electroretinography (ERG).The retinal morphology change was investigated with H&E staining.All of the results were quantified and statistically analyzed.Results In the control group,the amplitudes of a and b wave in the rod response were (18.33 ±3.53) μV and (107.58 ± 14.72) μV,while (64.80 ±7.57) μV and (178.76 ± 14.47) μV for the amplitudes of a and b wave in the maximum response;After treated by 6 months of intensive light exposure,in experimental group mice,the amplitudes of a and b wave in the rod response were (17.92 ±4.89) μV and (21.83 ± 5.51) μV;While in the maximum response a striking decrease was detected with a wave (18.23 ±4.44) μV and b wave (24.50 ± 4.49) μV,by compared with control group,the difference were statistical significant (all P ＜ 0.05).Histopathological analysis found significant loss of outer nuclear layer,photoreceptor out segment,whereas controls remained little change in the retina.And the retinal thickness decreased significantly from (181.32 ± 13.47) μm in control group to (102.34 ±9.38) μm after light insults in experimental group,the difference was statistical significant (P =0.017).Conclusion Intensive light exposure can cause the retinal structural and functional disorder in the AP-Pswe/PS1 transgenic mouse.