AIM: The expression of the mouse ? 2 microglobulin (? 2 m) in NIH3T3 cells transfected with the mouse ? 2m sense and antisense RNA was detected to clarify the effect of mouse ? 2m sense and antisense RNA on the expression of MHC classⅠgene. METHODS: The mouse sense and antisense RNA, pcDNA3-? 2mSN and pcDNA3-? 2mAN, were constructed and were transfected into NIH3T3 cells by lipofectamine. RT-PCR and Western blot were used to detect the expression of ? 2m in those cells. RESULTS: The expression of the mouse ? 2m in the cells transfected with pcDNA3-? 2mSN was increased, while it was decreased in those cells transfected with pcDNA3-? 2mAN. CONCLUSION: pcDNA3 -? 2mAN can downregulate the expression of the ? 2m in NIH3T3 cells.

AIM: The purpose of this study was to establish a new strategy for constructing the mouse ? 2m gene targeting vector in order to increase the homologous recombination frequency in contrast with our previous one, which was successfully constructed in the normal way.METHODS: A 4.2 kb 3' arm and a 0.8 kb 5' arm were amplified by PCR from the mouse ? 2m-pSV2△HXgpt genomic clone. They included the start region and the three exons, which were separated into two parts from exons 2 (the main coding block) for the two arms——5' arm and 3' arm.RESULTS: The two fragments, in reverse orientation to the Neo gene, were cloned into pPNT respectively on either side of Neo. They were identified by PCR, restriction analysis and sequence analysis as well. CONCLUSION: The mouse ? 2m gene targenting vector has been cloned successfully.

Objective To investigate the cause and related factors of hospitalization in patients with diabetic nephropathy with maintenance hemodialysis.Methods Fifty-six patients of diabetic nephropathy were enrolled in current study who hospitalized in Beijing Tongren Hospital affiliated to Capital Medical University from March 2009 to February 2012.All subjects were divided into hospitalized group (n =29) and non hospitalized group (n =27).The information including age,dialysis age,blood pressure,hemoglobin,blood glucose,serum creatinine,serum potassium,plasma albumin,serum calcium,serum phosphorus,albumin,intact parathyroid hormone (iPTH) and hemodialysis sufficienc (KT/V) were collected.Stepwise multiple regression analysis was applied to explore the main causes and related risk factor.Results Twenty-nine out of 56 patients were hospitalized and they had 56 hospitalization records during 24 months of observed period.Average hospitalization was 1.0 times per patient and hospitalized days were (29.62 ± 14.30) days per hospitalized patients.The main causes of hospitalization were cardiac vascular diseases (35.7％),vascular access stenosis (30.4％),and pulmonary infection (17.8％).Compare with non-hospitalization group,age of patient hospitalization group was older((66.97 ± 9.94) vs.(57.42 ± 11.77),t =2.917,P =0.02) and the level of albumin and Kt/V were lower(albumin:(34.02 ± 4.76) vs.(38.93 ± 3.82),t =-4.573,P =0.000 ; Kt/V:(1.16 ± 0.09) vs.(1.39 ± 0.18),t =6.503,P =0.000).Stepwise multivariate regression analysis showed plasma albumin and KT/V were risk factors to influence the hospitalization in diabetic patients undergoing maintenance hemodialysis (β =-1.724,-0.040 respectively,P =0.00,0.02 respectively) Conclusion Cardiovascular disease,vascular access stenosis and pulmonary infection were the important factors for hospitalization in hemodialysis patients with diabetic mellitus.Cardiovascular disease was the top reason of hospitalization in diabetic patients with maintenance hemodialysis.Hypoalbuminia and inadequacy dialysis were risk factors to influence the hospitalization in diabetic patients undergoing maintenance hemodialysis.

Objective:To measure the voice samples of the normal aged in order to systemically study the fea-tures of the voice changes. Method :To collect and analyze 146 voice samples of the normal aged with sonogram.Result:The fundamental frequency of the voice of the aged decreases and rises in the male more than 80 yearsolder. The low frequency harmonics are regulation and the intensity is strong in the formant of the aged. Thedifference reduces in voice between male and female. The harmonics to noise ratio tends downwards and the am-plitude perturbation quotient tends upwards along with the growth of age in the aged male. The changes of the above-mentioned parameters are not significant in the aged female. Conclusion:The voice changes are normalphysiological ones in the normal aged. The changes of the parameters are used to evaluate normal aged voice andabnormal one. The changes show that the function in the aged phonation tends to decline to a certain extent andit must be protected and be trained.

Child ; Cryptosporidiosis ; complications ; Cryptosporidium parvum ; Cyclospora ; Cyclosporiasis ; complications ; Diarrhea ; complications ; Humans ; Hypoproteinemia ; complications ; Male

Objective To explore risk factors of unintentional injury among left-behind children in a rural poverty county and to provide evidence for injury intervention.Methods 2 909 students,including 1 630 left-behind children,were recruited from a poor counties of five schools by Multi-stage stratified cluster random sampling method,and were surveyed applying designed questionnaire,interview method and observation method to collect the data of the injury accidents and related factors in the last one year.Results The incidence of unintentional injury among the left-behind children in rural area was 34.0％,higher than the rate of 27.7％ among non-left-behind children and the difference was statistically significant (c2 =13.541,P＜0.05);in the top five injury type were falling injury (14.0％),burus (5.6％) and sharp instrument injury (3.6％),bite (3.0％),parts (1.6％) among left-behind children.Low age,often miss my mother,fighting in school,outdoor activities,road and traffic state were risk factors for injury of left-behind children.Corcltsion The unintentional injury of left-behind children has become a social problem that can not be ignored,and targeted intervention should be carried out.

The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.

Objective:To prepare rabbit polyclonal antibody against human argonaute 3(AGO3) protein,to identify its properties and investigate the tissue distribution of AGO3 using tissue array.Methods:AGO3 peptide was synthesized using chemical method,and then conjugated to Keyhole limpet hemocyanin(KLH) as immunogen.The AGO3-KLH conjugate were injected into rabbits subcutaneously to produce polyclonal antibodies.The specificity and sensitivity of antibodies were identified by ELISA and Western blot after purification using affinity chromatography.Then the distribution of AGO3 in tissues was examined through immuno-stainning by tissue array.Results:Rabbit polyclonal antibodies against AGO3 were raised after immunization with AGO3-KLH conjugates.The anti-serum titer after the last inoculation was up to 1∶20 000.The preparations of the antibody were confirmed to raised recognize AGO3 peptides specially by ELISA and Western blot.AGO3 protein was stained positively in the cytoplasm of tumor cells and epithelial cells in many normal tissues.Conclusion:The polyclonal antibody against AGO3 protein has been achieved successfully,and it provides an efficient tool for further studying the roles of AGO3 in the pathway of miRNA and RNA interference and in the pathogenesis of human disease.

Objective To investigate the effects of angiotensin Ⅱ (AngⅡ) on the expression of albumin and the synthesis of type Ⅰ collagen in human normal hepatic cells. Methods HL-7702 cells (human normal hepatocyte) were cultured and divided into control group, Ang Ⅱ treated group, an AngⅡ+irbesartan (co-stimulated) group. The expressions of albumin and type Ⅰ collagen were detected by immunofluorescence and Western blotting, respectively. The mRNA level of type Ⅰ collagen was measured by real time-PCR(qRT-PCR). Results After stimulated with 10-7 mol/L Ang Ⅱ for 72 hours, the expression of albumin significantly decreased in Ang Ⅱ treated group compared with control group (0.85±0.11 vs 1. 41±0.23,P=0.000), while the mRNA expression increased in AngⅡ treated group compared with control group (1.00±0.08 vs 3.72±0.19,P=0.000). In costimulated group, however, the expression of albumin significantly increased (0.85 ± 0.11 vs 1.38 ±0.32,P=0.000),and mRNA expression (3. 72±0.19 vs 2.86±0.13,P=0.000) and synthesis of type Ⅰ collagen were reduced when compared with Ang Ⅱ treated group. Conclusions The reduction of albumin and elevated systhesis of type Ⅰ collagen in HL-7702 cells are induced via Ang Ⅱ AT1 receptor.

AIM: To investigate the feasibility and effect of directly differentiation of embryonic stem cells(ESC) into neural cells induced by retinoid acid(RA) without embryonic body(EB) culture period in vitro.METHODS: ESC were digested and divided into 4 groups: group A and B were undergone EB culturing.After that,cells in group A were induced by RA,cells in group B were differentiated spontaneously,cells in group C were committedly induced by RA directly without EB culturing,and cells in group D were differentiated spontaneously without EB period.Morphologic changes were observed under inverted microscope and scanning electron microscope.MAP-2 and GFAP were detected by immunocytochemistry and flow cytometry after differentiated for 9 days.RESULTS: In groups A or C,neuron-like cells increased gradually,forming neural network.At the 9th day,a large part of cells in these groups were MAP-2 positive cells,and the positive rate was higher than that in groups B or D(P0.05).CONCLUSION: ESC was directly induced into neural cells by RA without EB culture period in vitro.This modified method has the same effect as the traditional RA 4-/4+ assay and can replace the traditional method.