Objective To investigate the expression of CD4 +CD25 +CD127 low regulatory T cells in peripheral blood mononuclear cells( PBMNCs) in order to explore the influence on the immune status and disease progression in the different period of acute ischemic stroke. Methods 60 patients with acute cerebral infarction were selected as the stroke group,and divided them into the 24~48 h group (n=16), 3~7 d group (n=22) and 8~14 d group (n=22);22 healthy human were set as the control group. To analyze the percentage of CD4 +CD25 +CD127low regu-latory T cells in the peripheral blood of acute cerebral infarction patients and healthy human with flow cytometry. Results The percentages of CD4 +CD25 +CD127 low and CD4 +CD25 high regulatory T cells in the peripheral blood of the stroke group were significantly decreased at 24 ~48 h,3 ~7 d ( P <0.01 ) when compared with the control group, and then tended to return toward the control value at 8~14 d (P>0.05);the percentage of CD4 + regula-tory T cells in the peripheral blood of the stroke group was significantly higher than control group ( P <0.05 ) . Conclusion Imbalance of regulatory T cells is very likely to play an important role in the immunological injury of acute ischemic stroke. Regulatory T cells may be involved in the pathogenesis of acute ischemic stroke,and early detection may provide a basis for treatment.

[ ABSTRACT] Diabetes is characterized by an absolute or relative deficiency inβ-cell mass, which cannot be re-versed with existing therapeutic strategies.The restoration of the endogenous islet β-cells can stabilize the level of blood glucose.The isletβ-cells can be obtained from the directional differentiation of stem cells, but the process is complex and has the risk of teratomas generation.Cell direct reprogramming, one terminal differentiated cell can transdifferentiate into another kind of terminal differentiated cell, which is other than directional differentiation from stem cells.Direct reprogram-ming gives rise to the generation of isletβ-cells from one terminal differentiated cell, may be preferable for diabetes therapy because of its unique advantage.

ObjectiveTo determine the current students' social anxiety status and maternal factors in school students( grade 5-7) in Harbin,and provided scientific guidance relevant to students'social anxiety.Methods 1526 mother-child pairs collected by random cluster sampling,children completed the social anxiety scale for children (SASC),security scale,coping strategies questionnaire( consist of ambivalence and avoidance subscale),acceptance scale and psychological control scale,and mothers completed the center for epidemiologic studies depression inventory (CES-D),self-rating anxiety scale (SAS).ResultsThe total positive detection rate of social anxiety was 13.6％,there was no significantly difference between boys and girls (P ＞ 0.05) ; the positive detection rate of Grade 6 was higher than Grade 5 and Grade 7.There were positive correlation between social anxiety and mother's psychological control,mother's depression,mother's anxiety ( r =0.228,0.143,0.122) ; negative correlation with mother's acceptance( r =-0.214).Linear regression indicated that only three variables (mother's psychological control,mother's acceptance,mother's depression) were at the P ＜ 0.05 level.The regression coefficients for the three variables were 1.557,- 1.092 and 0.040,respectively.ConclusionMaternal highly psychological control,lowly acceptance and maternal anxiety depression all play a role in promoting the occurrence of students' social anxiety.

To investigate the effects of cembrane-type diterpenes extracted from Sinularia flexibilis on the proliferation of PC12 cells and their protective effects on PC12 cells exposed to glutamate.

Objective To explore the reliability of Chinese simplified diagnostic method for acute drug‐induced liver injury (DILI) in diagnosis of acute DILI .Methods From 2008 to 2013 ,a total of 320 patients diagnosed with acute DILI were enrolled .The clinical data of them were collected .International recognized Roussel Uclaf causality assessment method (RUCAM ) was taken as control and then simplified diagnostic method for DILI in China was evaluated . Variance analysis was performed for statistical analysis .Gamma value of two diagnostic methods was calculated and the correlation was analyzed .Results Among the 320 patients with acute DILI ,according to RUCAM ,there were 39 cases (12 .19% ) with quite high probability ,193 with high probability (60 .31% ) ,74 with possibility (23 .12% ) ,11 with less possibility (3 .44% ) and three with no probability (0 .94% ) .According to simplified diagnostic method for acute DILI ,194 cases were diagnosed (60 .62% ) ,103 were suspicious (32 .19% ) and 23 were excluded (7 .19% ) .The RUCAM score of diagnosed group (7 .5 ± 1 .2) was higher than that of suspicious group (5 .3 ± 1 .3) and excluded group (2 .1 ± 1 .1) ,and the difference was statistically significant (F =239 .545 ,P< 0 .01) .The correlation analysis between these two diagnostic methods indicated that Gamma value was 0 .955 (P < 0 .01) .Conclusions The simplified diagnostic method for acute DILI in China is simple ,practical and consistent with RUCAM .It can be used as one of the clinical methods for screening acute DILI .

BACKGROUND:Macrophage migration inhibitory factor is involved in the process of a variety of diseases, and plays a very important role in the tumor, autoimmune diseases, inflammation, angiogenesis, fibrotic diseases and so on. These biological characteristics are similar to keloids. OBJECTIVE: To compare the distribution and number of macrophage migration inhibitory factor in normal skin, hypertrophic scar and keloid. METHODS: We colected 40 clinical pathological scar specimens after surgery, including 20 hypertrophic scars and 20 keloids. Another 10 samples of the normal skin were used as control group. Hematoxylin-eosin staining and immunohistochemistry staining were performed to test the expression of macrophage migration inhibitory factor in pathological scars and normal skin. RESULTS AND CONCLUSION:Macrophage migration inhibitory factor was positively expressed in the normal skin, hypertrophic scar and keloid, and the expression of macrophage migration inhibitory factor in keloid was significantly higher than that in hypertrophic scar and normal skin (P < 0.01). It means that the abnormal infiltration of macrophage migration inhibitory factor may be associated with the formation of keloid.

Objective:To investigate the effect of piggyBac transpon,as a carrier of four defined transcription factors Oct4,Sox2,Klf4 and c-Myc,in the reprogramming of mouse embryonic fibroblasts (MEFs)to induced pluripotent stem cells (iPSCs).Methods:The MEFs were isolated from Oct4-GFP fetal mice and transfected by piggyBac transposon with four factors (Oct4,Sox2,Klf4 and c-Myc).The morphological changes of clones were traced with microscope during the process of induction.The chromosomes were analyzed to evaluate the karyotypic variation of iPSCs.The mRNA expressions of Oct4, Nanog and FGF4 associated with embryonic stem cells (ESCs)in the iPSCs of mice were tested by RT-PCR;the protein expressions of SSEA-1,Nanog and Alkaline phosphatase in the iPSCs of mice were determined by flow cytometry,immunofluorescence and AP staining.The iPSCs were transplanted into the NOD-SCID mouse groin,4 weeks later,the teratomas were removed for HE staining and the differentiation of tissue was observed.Results:The iPSCs were successfully obtained from MEFs by piggyBac carrying Oct4,Sox2,Klf4,and c-Myc.The round or oval iPSCs clones were similar to ESCs with clear boundry and large dense nuleus.The iPSCs showed the normal karyotypic and expressed the marker genes (Oct4,Nanog and FGF4)and proteins (SSEA-1,Nanog and AP)of ESCs.Teratomas containing three germ layers were formed in NOD-SCID mice after tanspalantation of iPSCs.Conclusion:The iPSCs are reprogrammed from MEFs by piggyBac transposon with four transcription factors-Oct4,Sox2,Klf4 and c-Myc,and the iPSCs with normal karyotype possess the characteristics of ESCs.

Objective To determine the development characteristics of social anxiety of peasant-worker's children in Harbin,and explore the effect of gender,parents quarrel,parents divorced and maternal factors on social anxiety.The study provided a scientific basis for the intervention of social anxiety.Methods The children in grade 1-5 from 3 schools in Harbin were collected by random cluster sampling in 2012.Then the same questionnaire was used to track the peasant-worker's children in 2014.The questionnaire included general information,Social Anxiety Scale for Children (SASC),Egna Minnen av Bardndosna Uppforstran (EMBU),Acceptance Subscale and Psychological Control Scale.Results (1) The detection rates of social anxiety of peasant-worker's children in 2012 and 2014 were respectively 29.3％,30.2％,and the detection rates had no significant difference (x2=0.111,P=0.790).But there was significantly difference between the two years(2012:6.90±4.26;2014:5.88±4.20;t=3.419,P=0.001).Children of newly occurring social anxiety in high grade was fewer and the ability to symptom relief was better.There was no significant difference between boys and girls on newly occurring and symptom relief(newly occurring:x2=0.018,P=0.910;symptom relief:x2 =0.541,P=0.518).(2) In 2012,the differences of the detection rate of children's social anxiety on parents quarrel,parents divorced were not significant (parents quarrel:x2 =0.881,P=0.393;parents divorced:x2=0.246,P=0.658).In 2014,the detection rate of children's social anxiety on parents quarrel,parents divorced were significantly higher than children in normal families (parents quarrel:x2=11.758,P=0.001;parents divorced:x2 =9.907,P=0.004).The detection rate of children's social anxiety of divorced parents increased with age(x2 =4.398,P=0.041).(3) In 2012 and 2014,children' s social anxiety had positive correlation with mother excessive interference and over-protective,mother refusing and denying,mother punishment,and had negative correlation with mother acceptance (P＜ 0.05).In 2014,it had positive correlation with mother control and negative correlation with mother emotional warmth understanding (P＜0.05).(4) In 2012,the regression coefficient of mother refusing and denying was 0.186.In 2014,the regression coefficient of mother acceptance and control were-0.266,0.169.Conclusion Social anxiety can be spontaneous remission,and the ability was better in high grade.Parents quarrel,parents divorced and maternal factors can affect the development of peasant-worker's children' s social anxiety,which should be concerned.

BACKGROUND:The smal intestinal submucosa has good biocompatibility and biodegradability, and also contains a variety of growth factors that can significantly promote celladhesion, proliferation and differentiation. Currently, the smal intestinal submucosa has been widely used in bone and cartilage, blood vessels, skin, bladder, smooth muscle and pancreatic tissue repair, showing good performance as a tissue-engineered cellscaffold.
OBJECTIVE:To investigate the in vitro feasibility of tissue engineered periosteum constructed by porcine smal intestinal submucosa and osteoblasts differentiated from bone marrow mesenchymal stem cells.
METHODS:Bone marrow mesenchymal stem cells were harvested from 2-week-old healthy New Zealand rabbits by using adherent method, and then cells were cultured, induced, differentiated and identified in vitro. Fol owing induced differentiation and identification, the bone marrow mesenchymal stem cells were compounded with porcine smal intestinal submucosa to fabricate tissue engineered periosteum. The adhesion, growth, and proliferation of cells on the materials were observed.
RESULTS AND CONCLUSION:At 5 days after inoculation, the cells receiving osteogenic induction could quickly adhere and proliferate on the surface of porcine smal intestinal submucosa and be interconnected;at 10 days, the desmosomes formed among the cells, cellprocesses from osteoblasts were visible and attached to the smal intestine submucosa;at 15 days, cellproliferation and secretion of matrix appeared, and multi-layer membrane-like structure formed on the surface of the smal intestine submucosa. These findings indicate that after osteogenic induction, the bone marrow mesenchymal stem cells can be combined with porcine smal intestinal submucosa to construct a tissue engineered periosteum, which is hoped to be an ideal scaffold for tissue engineering.

Objective To explore the potential role and function of miR-30 a in myocardial fibrosis after myocardi-al infarction( MI) .Methods We constructed the AAV plasmid vector which carried the miR-30 a gene of rat.The recombinant plasmid was detected by gene sequencing , enzyme digestion and PCR .Virus was packaged into HEK293 cells and virus titer was determined after extraction and purification by PCR .PBS fluid, rAAV9-miR-30 a-NC and rAAV9-miR-30 a were transmited to rat hearts from PBS group , miR-30 a-NC group and miR-30 a group respectively through transcoronary infusion before anterior descending coronary artery ligation .Sham group was set up at the same time.After 4 weeks, heart function was monitored by serial echocardiography , including fractional shortening ( FS) , and left ventricular ejection fraction ( LVEF) .Masson staining was used to calculate collagen volume fraction ( CVF) .The expression of collagen ⅠandⅢwere detected by immunohistochemistry . The mRNA level of miR-30a, TGF-β1 and CTGF were detected by real-time PCR.The protein level of TGF-β1 and CTGF were detected by western blot analysis .Results The cardiac function of miR-30 a group was improved significantly compared with PBS group and miR-30a-NC group (P<0.05).The levels of CVF,collagenⅠ,Ⅲexpression and Collagen Ⅰ/Ⅲ ratio in miR-30 a group was significantly lower than PBS group and miR-30 a-NC group ( P<0.01 ) .The mRNA and protein level of TGF-β1 and CTGF in miR-30 a group were reduced signifi-cantly than PBS group and miR-30 a-NC group ( P<0.001 ) .Conclusions The overexpression of miR-30 a after MI may reduce the mRNA and protein level of TGF-β1 and CTGF, so as to suppress myocardial fibrosis and im-prove cardiac function.