Objective To observe the effect of chronic cerebral hypoperfusion on abilities of learning and memory and astrocytes in old rats. Methods 50 Wistar rats were randomly divided into the sham group and model group with 25 animals in each group. All animals were assessed with Morris water maze to test the changes of abilities of learning and memory. The expressions of glial fibrillary acidic protein (GFAP) in the frontal lobe and hippocampus were observed immunohistochemically. Results Compared with the sham group, the scores of Morris water maze decreased in the model group, while the astrocytes marked by GFAP proliferated and enlarged significantly. Conclusion Proliferation and morphological changes of astrocytes are involved in pathological mechanism of chronic cerebral hypoperfusion, which might be associated with the decrease of ability of learning and memory.

Objective:To evaluate the influence of bone marrow stem cells ( BMSCs ) in the serological indicators of hepatolenticular degeneration combined liver fibrosis.Methods:60 cases were randomly divided into 3 groups: penicillamine group, BMSCs group and BMSCs+penicillamine group.BMSCs(2 ml)were injected into vein with normal saline(100 ml) every 10 days ( 3 times for a period of treatment).Liver tissue pathology biopsy was inspected and TBIL, ALT, ALB, CHE, PDGF-BB, TGF-β1, IL-6 and TNF-αwere detected at 0,4 ,8 and 12 weeks.Results: The level of serological indicators about liver functions were reduced in every group, while the changes in BMSCs+penicillamine group were especially obvious ( P<0.05 ).Conclusion: Penicillamine combined with BMSCs was effective in the improvement of liver functions of hepatolenticular degeneration combined liver fibrosis.

Objective To evaluate the effect of bone marrow stem cells (BMSCs) transplantation in the treatment of hepatolenticular degeneration of liver fibrosis. Methods Sixty cases with confirmed hepatolenticular degeneration of liver fibrosis were randomly divided into 3 groups: penicillamine group [40 mg/(kg·d)], BMSCs group and BMSCs + penicillamine group. Autologous BMSCs (2 mL) were injected into vein with normal saline (100 mL). Liver tissue pathology biopsy was inspected and the changes in HA, PCⅢ, LN, CⅣ, TIMP-1 and MMP-1 were observed at 0, 4th, 8th and 12th week during the therapeutic process. Results The level of serum fibrotic markers were reduced in every group , while the changes in BMSCs + penicillamine group were especially obvious (P < 0.05). Conclusion Penicillamine combined with BMSCs was effective in the therapy of hepatolenticular degeneration of liver fibrosis.

Objective To observe the survival and migration characteristics after intracarotid transplantation of bone marrow stem cells(BMSCs) and its effect on learning-memory abilities across the blood-brain barrier(BBB) in the vascular dementia(VD) rats .Methods Bone mononuclear cells(BMNCs) were isolated from bone marrow in vitro by standard Ficoll-Hypaque technique , then cells were enriched and expanded by using bone marrow adherent culture .72 Wistar rats were meanly divided into control group ,model group and treatment group .The VD rat model was established by modified pulsinellis 4-vessel occlusion(4 VO) .The treatment group received intracarotid infusion of 0 .5 mL 1 .2 × 107/mL BMSCs which were labeled with BrdU in vitro after opera-tion .Their survival ,migration and the learning-memory abilities were observed at 4th and 8th week .Results BMSCs transplanted by intracarotid transplantation survived and had been found throughout the brain tissue .They major migrated and localized in the is-chemic zone of injury such as hippocampus and cerebral cortex .Compared with the control group ,active avoidance response(AAR) ratio in the model group(42 .1 ± 4 .5) ,group(43 .6 ± 3 .6)showed significantly decrease compared with the control group (90 .0 ± 4 .3) ,(92 .5 ± 5 .0)(P<0 .01) ,and the treatment group(69 .2 ± 4 .7) ,(70 .8 ± 4 .7)was significant higher than the model group(P<0 .01) .Conclusion Intracarotid transplantation of BMSCs could enter the VD rats cerebra parenchyma via BBB ,migrate into the damaged brain tissue to gather and survive .Learning-memory abilities can be improved significantly by transplanted BMSCs .

Objective To investigate the changes of learning and memory function ,vascular endothelial growth factor (VEGF) and heme oxygenase‐1(HO‐1) expression in cerebral cortex and hippocampus of chronic ischemic vascular dementia rats . Methods Thirty‐six healthy SD rats were divided into the control group ,sham operation group and model group ,12 cases in each group .The chronic ischemic vascular dementia rat model was established by the permanent bilateral carotid artery occlusion The sham operation group received the same treatment to the model group except without bilateral carotid artery occlusion .The learning and memory abilities were tested by the Morris water maze experiment and climbing rope strength experiment at 1 ,4 ,8 ,12 weeks respectively .The expressions of VEGF and HO‐1 in rat cerebral cortex and hippocampus was determined by immunohistochemical SP technique .Results The escape latency time at 8 ,12 weeks in the model group was longer than that in the sham operation group and control group ,and the number of crossing the platform was less than that in the sham operation group and control group ,the differences were statistically significant (P<0 .05);the time of climbing at 1 -12 weeks had no statistical difference between the model group and the sham operation group and between the model group and the control group (P>0 .05) .The positive expression of HO‐1 and VEGF protein contents in the control group and sham operation group was less than that in the model group with sta‐tistical difference(P<0 .05) .Conclusion Chronic cerebral hypoperfusion has a permanent damage to the learning and memory abil‐ities in rats ,while has no influence on the motor function .VEGF and HO‐1 may play a protective role in chronic cerebral ischemia .

Bone marrow mesenchymal stem cells (BMSCs),which can be isolated from organs and tissues,are cell populations with high degree of plasticity,similar to hematopoietic stem cells in bone marrow,and can be in vitro cultured,induced and amplified.BMSCs are increasingly used in gene therapy,cell therapy and tissue engineering.BMSCs have been currently studied in a number of autologous transplantations,while not all BMSCs are suitable for autologous transplantation.BMSCs exhibit biological aging gradually when cultured in vitro.Biological aging can be divided into age-induced senescence and long-term passage induced senescence.Aging BMSCs demonstrate changes in biological behaviour which reduces the success rate of autologous transplantation.In this review,biological aging BMSCs are discussed in order to provide help for the transplantation of BMSCs.

BACKGROUND:The microRNAs are involved in regulation of stem cel proliferation, differentiation and aging. To study the effect of Let-7c, a member of Let-7, on the neural differentiation of bone marrow mesenchymal stem cels provides new ideas for stem cel therapy. OBJECTIVE: To investigate the role of Let-7c in the neural differentiation of bone marrow mesenchymal stem cels. METHODS: The lentiviral vectors of Let-7c-up and Let-7c-inhibition were constructed and transfected into rat bone marrow mesenchymal stem cels. Optimal multiplicity of infection was screened. The cels were divided into non-transfected group, negative control group (transfected with empty virus), transfected enhancement group (transfected with LV-rno-Let-7c-up), transfected inhibition group (transfected with LV-rno-Let-7c-5p-inhibition). Bone marrow mesenchymal stem cels were treated with fasudil as an inducer for triggering the cels to differentiate into neurons. The fluorescence expressed by transfected cels was observed under inverted fluorescence microscope. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The cel viability was determined by MTT method. RESULTS AND CONCLUSION:Under the inverted fluorescence microscope, the cels were successfuly transfected with LV-rno-Let-7c-up and LV-rno-Let-7c-5p-inhibition. Fasudil induced bone marrow mesenchymal stem cels to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in the transfected enhancement group were significantly higher than those in the negative control group (P < 0.05), while in the transfected inhibition group, they were lower than those in the negative control group (P < 0.05). These findings indicate that the differentiation percentage of bone marrow mesenchymal stem cels is increased by fasudil after transfection with LV-rno-Let-7c-up, and Let-7c may promote the differentiation of bone marrow mesenchymal stem cels into neurons.

Objective To investigate cognitive impairment mechanism by studying dorsolateral prefrontal cortex connectivity in patients with primary insomnia.Methods Forty patients with primary insomnia and 50 healthy subjects from the Department of Neurology,People's Hospital of Zhengzhou University during the period April 2011 through April 2013 were included.The World Health OrganizationUniversity of California Los Angeles Auditory Verbal Learning Test (WHO-UCLA AVLT) and the digital pin test were applied to evaluate the subjects' word study ability and vigilance.Resting state functional magnetic resonance imaging was used to observe the connectivity of dorsolateral prefrontal cortex.Results The Pittsburgh Sleep Quality Index (2.00 (1.00,3.00)) and the Hamilton Anxiety Scale scores (13.00 (11.25,15.75)) of primary insomnia patients were significantly higher than that of healthy controls (11.00(9.00,13.00),1.00 (0,2.00),Z=-5.517,Z=-5.525,P＜0.01).Digital pin test efficiency (60.03％ ± 13.95％ vs 66.32％ ± 13.73％,t =2.142,P＜0.05) and WHO-UCLA word learning (10.11 ± 2.29 vs 11.95 ± 2.42,t =-3.493,P ＜ 0.01) of primary insomnia patients were significantly lower than that of healthy controls.Compared to the healthy controls,the right dorsolateral prefrontal cortex of primary insomnia patients exhibited decreased functional connectivity of the right prefrontal lobe (-2.610 3 ± 0.172 6,t =-3.504,P ＜ 0.05).The left dorsolateral prefrontal cortex of primary insomnia patients exhibited increased functional connectivity of the bilateral insular lobes and right prefrontal lobe (2.8204±0.326 5,2.371 7 ±0.106 6,2.492 6 ±0.052 8,t =4.032,t =3.340,t =3.037,P ＜0.05).Conclusions The ability of WHO-UCLA word study and the digital pin test efficiency have been shown to decline in patients with primary insomnia.The possible mechanism of cognitive impairment may be the abnormal dorsolateral prefrontal cortex connectivity in patients with primary insomnia.

Objective To analyze the etiological factor and genetic feature of a familial hemophagocytic lymphohistiocytosis patient with PRF1 mutation (FHL2) with human herpesvirus 7 (HHV7)infection and its family constellation. Methods Clinical characteristics, laboratory examinations of a FHL2 case with HHV7 infection were reported. HHV1-HHV8 virus DNA was screened by PCR; NK cell function was analyzed by flow cytometry; PRF1 gene mutations were analyzed by PCR and direct sequencing, structure of mutant PRF1 proteins were analyzed using ExPasy and I-TASSER server and genetics pedigree were analyzed. Results The patient's HHV7 viral was detected positive with DNA copy number of 350/106 peripheral nucleated cells. Flow cytometry analysis showed decrease both in proportion of perforin positive NK cells and perforin protein expression. Genetic testing showed PRF1 biallelic heterozygote mutations (c. 503G ＞ A/p. S168N and c. 1177T ＞ C/p. C393R) and pedigree analysis showed they were inherited. The patient was then treated with antivirus therapy, dexamethasone and VP16 therapy, but only achieved partial response. The patient was then followed by human leukocyte antigen 10/10 allele identical nonconsanguinity allogeneic hematopoietic stem cell transplantations (allo-HSCT) and soon the successful implantation of donor hematopoietic cells and persistent recovery was achieved. The patient was now surviving without recurrence for 9 months after allo-HSCT. Conclusions FHL is prone to be misdiagnosed as lymphoma. Genetic analysis of related gene mutation and herpes simplex virus detection will help in early and accurate diagnosis. Allo-HSCT is a fundamental treatment of FHL.

BACKGROUND:There is no clear understanding about the effect of let-7f and interleukin-6 (IL-6) on the proliferation of bone marrow mesenchymal stem cels and their relationship.
OBJECTIVE: To explore the effects of expression levels of let-7f and IL-6 on the proliferation of bone marrow mesenchymal stem cels and their relationship.
METHODS:(1) LV-rno-let-7f-up and LV-rno-let-7f-down were constructed and transfected into bone marrow mesenchymal stem cels of Sprague-Dawley rats, respectively. Then, there were four groups in the study: transfection upregulation group transfected with LV-rno-let-7f-up), transfection inhibition group (transfected with LV-rno-let-7f-down), negative control group (transfected with FU-RNAi-NC-LV), and untransfected group. The expression level of let-7f in each group was detected by qRT-PCR. The proliferation ability of cels and expression levels of IL-6 when let-7f expression was at different levels were detected by MTT, flow cytometry and ELISA. The expression of Cyclin D1 at mRNA and protein levels was detected by qRT-PCR and western blot, respectively. (2) To predict the potential target gene of let-7f, the wild-type/mutant IL-6 3’UTR reporter gene vectors were constructed, and cotransfected with let-7f/let-7f inhibitor respectively into the 293T cels to measure the luciferase.
RESULTS AND CONCLUSION: Compared with the negative control group, the proliferative and cloning capacities of cels in the transfection upregulation group were higher; the number of cels was significantly decreased at G1 stage and increased at S stage, and the apoptotic cels were reduced in number (P < 0.05). However, the transfection inhibition group had opposite results. The expression level of IL-6 in the transfection upregulation group was lower than that in the untransfected group and negative control group (P < 0.05); while in the transfection inhibition group, the expression level of IL-6 was significantly increased (P < 0.05). The expression of Cyclin D1 at mRNA and protein levels was up-regulated in transfection upregulation group (P < 0.05) and down-regulated in the transfection inhibition group (P < 0.05), but there was no significant difference between the negative control group and untransfected group (P > 0.05). Luciferase activity of cels transfected with wide-type IL-6 3’UTR and let-7f was significantly reduced (P < 0.05). These findings indicate that up-regulation of let-7f can promote the proliferative and cloning capacities of bone marrow mesenchymal stem cels and reduce cel apoptosis, but downrelation of let-7f exhibits an inhibitory effect. Overexpression of IL-6 can suppress the proliferation of bone marrow mesenchymal stem cels, which is considered to be a target gene of let-7f, and let-7f may suppress the expression of IL-6 to promote the cel proliferation.