AIM:To investigate the therapeutic effect of menstrual blood-derived endometrial stem cells(MenSCs)on chemotherapy-induced intestinal mucositis and flora disorders in mice,and to explore the potential mecha-nism.METHODS:The mice were randomly divided into 3 groups including normal treatment,cisplatin(Cis)treatment and Cis+MenSC treatment,with 10 mice in each group.To induce intestinal mucositis,the mice were treated with Cis(2 mg·kg-1·d-1)by intraperitoneal injection for 5 consecutive days.Control mice for normal group were received equal vol-umes of normal saline.For Cis+MenSC treatment,MenSCs(1×106)was transplanted into the mice of Cis treated mice through tail vein.The performances and weight changes of mice were examined during the experiment.After the treat-ment,the small intestine and colon were isolated for subsequent HE staining,the ratio of F4/80 and IL-6 positive cells in small intestine were detected by immunohistochemical staining,and the expression of tight junction,inflammation and apoptosis related proteins was detected by Western blot.16S rDNA amplicon sequencing was performed to detect the diver-sity and richness of intestinal flora in mice.RESULTS:Compared to the Cis group,the MenSCs-treated mice showed sig-nificantly increased body weight,relieved intestinal lymphocytes infiltration,alleviated intestinal villous edema,and or-derly arranged glands in intestinal tissues.Further analysis indicated that MenSCs transplantation significantly up-regulat-ed the expression of intestinal tight junction related proteins ZO-1 and occludin in Cis-treated mice(P＜0.05).Subse-quently,MenSCs transplantation significantly inhibited the macrophages infiltration in intestinal tissues(P＜0.01),down-regulated the expression of pro-inflammatory factors IL-1 and IL-6 and pro-apoptotic protein Bax(P＜0.01),while up-regu-lated anti-inflammatory factor IL-10 and anti-apoptotic protein Bcl-2(P＜0.01).Additionally,further microflora sequenc-ing indicated that MenSCs transplantation prevented mice from Cis-induced intestinal flora disorder,and significantly re-duced the abundance of harmful bacteria such as isenbergiella tayi and Anaerotruncus colihominis(P＜0.01).At the same time,the abundance of beneficial bacteria Lactobacillus apodemi was increased(P＜0.05),thereby restoring the composi-tion and function of healthy intestinal flora.CONCLUSION:MenSCs transplantation alleviates the chemotherapy-in-duced damage of intestinal structure,relieves the symptoms of chemotherapy-induced mucositis and restores the homeosta-sis of intestinal flora in mice.

Hippocampal formation is important in spatial learning and memory. Members of the cadherin superfamily are observed in the neural system with diverse spatial and temporal expression patterns and are involved in many biological processes. To date, the avian hippocampal formation is not well understood. In this study, we examined the expression of cadherin mRNA in chicken and mouse brains to investigate the morphological and cytoarchitectural bases of hippocampal formation. Profiles of the spatiotemporal expression of cadherin mRNAs in the developing chicken embryonic parahippocampal area (APH) are provided, and layer-specific expression and spatiotemporal expression were observed in different subdivisions of the APH. That fact that some cadherins (Cdh2, Cdh8, Pcdh8 and Pcdh10) showed conserved regional expression both in the hippocampus and entorhinal cortex of mice and the hippocampal formation of chickens partially confirmed the structural homology proposed by previous scientists. This study indicates that some cadherins can be used as special markers of the avian hippocampal formation.

Objective: To study the role of Treg cells in the development of mouse experimental autoimmune encephalomyelitis (EAE) through depleting or transplanting Treg cells. Methods: C57BL/6 mice were injected with anti-CD25 monoclonal antibody to deplete natural CD25-expressing Treg cells in vivo, and then treated with MOG_35-55/CFA to induce EAE. Their EAE scores were compared with those of the mice without Treg cell deletion (control group). The numbers and percentages of CD4⁺ Foxp3⁺ cells in mouse blood samples on 6 d, 10 d, 20 d and 35 d were quantified using flow cytometry. To evaluate the therapeutic effect of Treg cells transplantation on EAE, magnetic activated cell sorting (MACS) combined with fluorescence-activated cell sorting (FACS) was used to isolate Treg cells from spleen and lymph nodes of Foxp3^GFP+ transgenic mice on 6 d after EAE induction. Then the cells were injected through tail vein into wild-type mice on 6 d after EAE induction. The EAE scores of both recipient and control mice were recorded and compared. Results: The efficiency of natural Treg cells depletion with anti-CD25 antibody was above 95%. The mice with Treg cell depletion developed significantly more severe EAE than the control mice after MOG_35-55/CFA induction. FACS analysis of Treg cells during the development of EAE demonstrated that the lowest Treg cell percentage was detected on 6 d after EAE induction, hence it was the time point for the transplantation of Treg cells. CD4⁺ GFP⁺ Treg cells were isolated from Foxp3^GFP+ transgenic mice on 6 d after EAE induction and immediately transplanted into wild-type mice on 6 d after EAE induction. The transplantation of isolated Treg cells significantly alleviated the EAE in mice as compared with the control group. Conclusions Mice with Treg cell depletion developed severer EAE than the control mice after induction, but the EAE score could be significantly reduced with the transplantation of Treg cells. This study showed that the transplanted Treg cells had protective effect on mice during the course of EAE development. Thus, Treg cell transplantation could be used as an effective therapeutic approach for the treatment of multiple sclerosis (MS).

Objective To analyze the differences in immune system between Npc1 gene mutant (Npc1-/ -) and wild-type (Npc1+/ +) mice for better understanding the pathogenesis of Niemann-Pick disease type C1 (NPC1) from an immunological perspective and providing reference for NPC1 treatment in clinic.Methods Body, thymus and spleen weight of Npc1-/ -and Npc1+/ + mice aged (14±2) days, (42±2) days and (63±2) days (Day14±2 , Day42±2 and Day63±2 ) were recorded and the associated organ index were calcu-lated. White blood cell count in peripheral blood of mice aged Day42±2 was examined by routine blood test. Expression of cytokines at mRNA level in mouse peripheral blood was detected by qPCR. Percentages of CD4+, CD8+ and CD19+ lymphocytes in peripheral blood and spleen of mice aged Day42±2 were measured by flow cytometry. Apoptosis and senescence of spleen in mice aged Day63±2 were examined by immunofluores-cence and β-galactosidase staining. Results Compared with Npc1+/ + mice, there was no significant differ-ence in the weight of spleen and thymus in Npc1-/ - mice aged Day14±2; the weight of spleen in Npc1-/ - mice aged Day42±2 significantly increased, but the weight of thymus showed a significant decrease; furthermore, both the weight of spleen and thymus in Npc1-/ - mice aged Day63±2 significantly decreased; and the body weight of Npc1-/ - mice of each age group significantly decreased. Moreover, compared with Npc1+/ + mice, the absolute number of lymphocytes in the peripheral blood of Npc1-/ - mice aged Day42±2 showed no signifi-cant difference, but the percentage in whole white blood cells significantly decreased due to the significantly increased neutrophils. Expression of cytokines ( IL-1, IL-2, IFN-γ, TNF-α, IL-4, granzyme A and granzyme B) at mRNA level in the peripheral blood leukocytes of Npc1-/ - mice aged Day42±2 was abnormal as compared with that in Npc1+/ + mice. The number of T (CD4+ and CD8+) lymphocytes in Npc1-/ - mice aged Day42±2 significantly decreased, while the number of B (CD19+) lymphocytes increased significantly as com-pared with those in the Npc1+/ + mice. Compared with Npc1+/ + mice, apoptosis and senescence of the spleen in Npc1-/ - mice aged Day63±2 aggravated significantly. Conclusion The abnormal lipid metabolism triggered by Npc1 gene mutation causes severe immune dysfunction in Npc1-/ - mice. Therefore, immune dysfunction should be taken into full consideration when treating patients with NPC1, which might help improve the life quality and prolong the survival time.

Objective To investigate the growth curve,breeding rate,and blood physiological and biochemical parameters in Npc1 gene mutant mice (Npc1-/-) for providing theoretical evidence in research on Niemann-Pick disease type C1 (NPC1) patient.Methods 1) The body mass of Npc1-/-,Npc1+/-,and Npc1+/+ mice (n=120;60♀,60♂) was measured from 0 to 77 days;(2) As Npc1-/-mice were born only by the mating Npc1+/-mice,the breeding rate of Npc1+/-mice was counted here from the 1st to 4th generation;(3) The blood physiological and biochemical parameters were measured on both Npc1-/-and Npc1+/+ mice at 60 days.Results 1) Compared with the wild type controls,the body weight of Npc1-/-mice was progressively increased up to 7 weeks and then decreased,and died around 11 weeks.The body weight of the Npc1+/-and Npc1+/+ mice was increased as time went on.After 4 weeks,the male mice showed a higher weight gain than the females;(2) The generations of Npc1+/-mice had no significant difference in mating-parturition interval,litter size,weaning litter and the number of male and female (P>0.05),but the weaning rate of the 2nd generation was significantly higher than that of the 1st generation (P<0.05);(3) The hematological parameters showed a significant difference only in mean corpuscular hemoglobin (MCH) and mean peroxidase index (MPXI) between the Npc1-/-and Npc1+/+ mice (P<0.05).No significant difference was found in other hematological parameters (P>0.05).Among the biochemical parameters,aspartate aminotransferase (AST),glucose (GLU),lactate dehydrogenase (LDH),potassium (K) and copper (Cu) had a significant difference between the Npc1-/-and Npc1+/+ mice (P<0.05).Conclusions 1) The growth curves of Npc1-/-,Npc1+/-,and Npc1+/+ mice are different due to different genotype and sex;(2) The reproduction rates of Npc1+/-mice have no significant difference among different generations;(3) The blood physiological parameters (MCH,MPXI) and biochemical parameters (UREA,AST,GLU,LDH,K,Cu) are significantly different between Npc1-/-and Npc1+/+ mice.

Brain-computerinterface(BCI) is a kind of direct channel for information communication and control established between the human brain and computer or other electronic equipment.BCI is a novel information communication system which does not depend on the conventional brain information pathways.The asynchronous brain-computer interface technology is based on alpha wave control,and can automatically switch system mode between working and idle and select the larger EEG signal associated with motion imagination.In this paper,the basic knowledge of BCI and alpha wave-based asynchronous BCI technology were introduced.The key technology and application prospect of the novel alpha wave-based asynchronous BCI technology were summarized,and the status and existing problems were analyzed.

AIM:To investigate the renal function and pathological changes in Npc1 mutant ( Npc1-/-) mice. METHODS:Different genotypes of Niemann-Pick disease type C1 (Npc1) mice were identified by PCR.Subsequently, the renal function of Npc1-/-and Npc1 +/+mice at postnatal day 60 ( P60) was evaluated by measuring the activity and con-tent of important indicators in the serum including ALT , AST, LDH, urea, UA and Cr.Furthermore,β-galactosidase stai-ning and Masson staining were performed to examine the aging and fibrosis of the renal tissues , respectively .RESULTS:Compared with the Npc1 +/+mice, the body weight and kidney weight had a significant reduction ( P<0.01) in the Npc1-/-mice.The results of hepatic and renal functions showed that the activities of ALT , AST and LDH, and contents of urea, UA and Cr had marked increases (P<0.05) in the Npc1-/-mice.Moreover, the results of senescence-associatedβ-galacto-sidase staining in the renal tissues demonstrated accelerated aging in the Npc1-/-mice (P<0.01), and these results were confirmed by Masson staining, which clearly showed the formation of collagen fibers (P<0.01).CONCLUSION:Muta-tion of the Npc1 gene results in abnormal lipid metabolism , which accelerates kidney senescence by promoting fibrosis in the renal tissue and subsequently causes reduction in renal function .

[Abstract ] In recent years, induced pluripotent stem cells (iPSCs) technology has played an important role in basic and clini-cal application research of Parkinson′s disease ( PD) and acquired significant progress .The neural progenitor /stem cells or dopamine ( DA) neurons which were obtained through iPSCs technique and direct differentiation technique from somatic cells were used for the study of cell therapy in PD , and good results were achieved .The cell models of DA neurons were established from PD patients carrying LRRK2, PAKK2, PINK or SNCA mutations via iPSCs technology , and the mitochondrial function and morphology , oxidative stress,α-synuclein ( SNCA) accumulation , and other aspects were studied on the pathogenesis of PD .This article briefly reviews the latest pro-gress of iPSCs technology in transplantation for treatment of PD and the establishment of cell model of PD disease , and provides refer-ence for further research .

Objective:To explore the role of N-cadherin and β-catenin in the formation of spinal commissural axon projection during chicken embryonic development.Methods:Fertilized eggs were cultured for three days(stage22),N-cadherin orβ-catenin inter-ference plasmid was injected into the neural tube and in vivo electroporation was performed.Three days after the electroporation, embryos were collected,fixed with 4%PFA,embeded with OCT,and cut into frozen sections.Four groups ( knockdown of N-cadherin orβ-catenin or both of them,and control) were included in this study.Immunohistochemistry method was used to analyze the protein expression result of N-cadherin or β-catenin.The changes of spinal commissural projections were observed with GFP fluorescence.Results:During chicken embryonic development,knockdown of N-cadherin inhibited the expression of β-catenin in the spinal cord.The commissural nerve fibers projecting to the contralateral side of the spinal cord was impaired after knockdown of N-cadherin or β-catenin;this phenotype was similar after knocking down both of them.Conclusion: N-cadherin is implicated in the formation of spinal commissural projection in the developing spinal cord,possibly viaβ-catenin.

BACKGROUND:Human glial cellline-derived neurotrophic factor (GDNF) and vascular endothelial growth factor 165 (VEGF 165 ) are essential genes for celldifferentiation.
OBJECTIVE:To construct and identify pIRES 2-GDNF-VEGF 165 bicistronic eukaryotic expression vector.
METHODS:Human GDNF genes were obtained from the genomic DNA of human peripheral blood mononuclear cells by PCR. Then the GDNF cDNA fragment was inserted into the multiple cloning sites of pIRES 2-EGFP, to generate the bicistronic eukaryotic expression plasmid pIRES 2-GDNF-EGFP. The VEGF 165 gene was obtained from pIRES 2-VEGF 165-EGFP plasmid by twin PCR. Then VEGF 165 cDNA fragment was cloned into the pIRES 2-GDNF-EGFP, instead of EGFP, to create a double gene co-expressing vector plasmid pIRES 2-GDNF-VEGF 165 containing internal ribosome entry sites. Then pIRES 2-GDNF-VEGF 165 was used to transfect HEK293 cells. RT-PCR and western blot analysis were performed to test the co-expression of double genes.
RESULTS AND CONCLUSION:DNA sequencing analysis demonstrated that the GDNF and VEGF 165 were exactly consistent with the sequence recorded in the GenBank. The size of GDNF gene was 636 bp and the size of VEGF165 gene was 576 bp. Enzyme digestion analysis indicated that, pIRES2-GDNF-VEGF165 bicistronic eukaryotic expression vector inserted GDNF band by Bgl II/Bam HI, inserted IRES-VEGF 165 fragment by Bam HI/Not I, and inserted GDNF-IRES-VEGF165 fragment by Bgl II/Not I. RT-PCR and western blot analysis showed that, after HEK293 cells were transfected with pIRES 2-GDNF-VEGF 165 , double genes were expressed at the mRNA and protein levels. The pIRES 2-GDNF-VEGF 165 bicistronic eukaryotic expression vector is successful y constructed.