Objective To investigate the relationship between diabetic retinopathy(DR) and the state of diabetes mellitus(DM) control.Methods The factors for the occurrence and development of DR were analyzed by non conditional Logistic regression model.Results 5 significant factors were screened out,which were urinary microalbumin excretion index(ALB/Cr),blood glucose before breakfast,glycosylated hemoglobin(HbAlc),diabetic neuropathy and course of DM.And the sugestible threshold value of blood glucose before breakfast is 7.90mmol/L at which we can prevent the occurrence of DR,and at 7.15mmol/L,occurrence of DR can be strictly controlled.Conclusion The state of controlling blood glucose and other complications in DM patients influences the occurrence and development of DR.

Objective To Summarize the application of digital artery dorsal distal interphalangeal joint perforator flap to repair finger tip skin defect,and at the same time,the method and effect of anastomosis of finger dorsal branch of digital nerve sensory reconstruction.Methods From September 2012 to March 2015,78 patients 92 fingers were treated in Orthopedics Department of Jinshan Branch Hospital of the Sixth People′s Hospital of Shanghai,all patients with finger artery distal interphalangeal joint dorsal cutaneous branches of the dorsal flap pedicled with retrograde,transferred to repair the skin defect of finger end,and anastomosis of dorsal branch of the proper digital nerve reconstruction.Results Postoperative vascular crisis occurred in 8 cases,2 cases of partial flap necrosis and healed after symptomatic treatment.All flaps survived,the wounds healed in I stage,and the donor site healed in I stage.All the 73 cases were followed up,the follow-up period ranged from 3.0 to 12.0 months,an average of (7.8±2.5) months.The postoperative appearance and feel good,soft texture,abrasion resistance,no tenderness,cold resistance,dynamic two-point recovered to 4.0-8.0 mm,average (5.3±0.9) mm.Static two-point discrimination was 4.0-9.0 mm,average (5.8±1.2) mm.The method of TAM was used to determine the function of the 67 fingers,good for the 7 finger,but also for the 5 finger and the difference of the 0 finger.The excellent and good rate was up to 93.7%.Conclusion The operation without sacrifice of major arteries and nerves by finger,the middle finger dorsal skin for non functional surface area,and at the same time by anastomosis of dorsal branch of the proper digital nerve reconstruction,without surgery two times,is one of the ideal surgical repair of skin defect of the finger end.

ObjectiveTo investigate the expression of miR-155 and miR-146a in peripheral blood mononuclear cells (PBMC) and plasma of rheumatoid arthritis (RA) patients.MethodsPBMC and plasma were separated from the peripheral blood of 34 RA patients and 15 healthy individuals.Total RNAs were isolated and miRNAs were purified.The levels of miR-155 and miR-146a were determined by quantitative reverse transcription PCR(qRT-PCR).U6 was used as housekeeping control.The amount of target miRNA was normalized relative to the amount of U6(ΔCt=ΔCtmiRNA-ΔCtU6).Relative expression levels were expressed as 2 △-ΔCt.Data were analyzed using SPSS 13.0 software.The test of homogeneity of variance and unpaired t-test was used to compare between groups.P values(2-tailed) less than 0.05 were considered as statistically significant.ResultsThe expressions of PBMC and plasma miR-155 were higher in RA patients than those in the healthy control individuals(0.08±0.08 vs 0.05±0.03,t=-2.225,P＜0.05; 5.9±6.7 vs 1.3±2.0,t=-3.677,P＜0.05).The expression of miR-146a in PBMC and plasma of RA patients and controls were(1.3±1.2 vs 0.8±0.6,t=-2.154,P＜0.05)and(741±1001 vs 300±295,t=-1.669,P＞0.05).According to their DAS28 value,RA patients were divided into high activity group (23 cases,DAS28≥5.0) and low disease activity group( 11cases,DAS28＜5.0).The plasma miR-155 and miR-146a expressions were significantly higher in high activity group than those in low activity group.There were no significant differences in the expression of PBMC miR-155 and miR-146a between the two groups.ConclusionThe expression of PBMC and plasma miR-155 and miR-146a are higher in RA patients.The expression of plasma miR-155 and miR-146a are associated with RA patients' activity.Plasma miR-155 and miR-146a may be potential non-invasive biomarkers for RA diagnosis anddisease activity assessment.

Objective To obtain related genes of Cysticercus cellulosae from spliced leader (SL) cDNA library. Methods Spliced leader library of Cysticercus cellulosae was constructed using SL specific primer and oligo (dT)15 with M13M4 primer, and positive clones were then screened randomly, identified with enzyme restriction, followed by sequencing and homologous analysis. Results The amino acid sequence, encoded by the positive clone with a poly (A) 22 tail and a complete open reading frame (ORF), was with homology of RNA polymerase subunit genes of human, B. napus, fission yeast, A. thaliana, C. elegans and fruit fly up to 71.6%. Conclusion The protein, RNA polymerase subunit encoded putatively by the clone, is high conservative in different species.

Objective To investigate the expression of miRNA-16 in peripheral blood monouclear cells (PBMC)from systemic lupus erythematosus( SLE)patients. Methods Sixteen SLE patients who meet the diagnostic criteria of SLE revised in 1997 American rheumatology and 12 healthy individuals were selected as our subjects. Their peripheral blood were sampled. Total RNAs were extracted and purified. The level of miRNA-16 was determined by quantitative reverse transcription PCR( qRT-PCR). U6 was used as housekeeping control. The amount of target miRNA was normalized relative to the amount of U6(ΔCt =ΔCt miRNA-ΔCtU6 ). Relative expression levels were expressed as 2-ΔCt . Results The expression level of miRNA-16 in the SLE patients was 919. 87 ± 715. 45,significantly higher than that in the healthy control group(413. 6 3 ± 330. 69;t= -2. 497,P﹤0. 05). And miRNA-16 expression in SLE active group was 1 298. 79 ± 803. 79,significantly higher than that in SLE stable group(540. 95 ± 350. 15;t= -2. 445,P﹤0. 05). The level of miRNA-16 was related with AnuA (r=0. 669,P=0. 005),ESR(r=0. 608,P=0. 012)and SLEDAI(r=0. 530,P=0. 035). Conclusion The expression of miRNA-16 is high in SLE patients and it is related with SLE activity.

Objective To elucidate the function way of micro RNA(miR)-155 in the differentiation of Th17 cells.Methods CD4+T cells were separated from mice spleens using MACS CD4+T cells separatinge kit and cultured with interleukins [interleukin (IL)-2, IL-23 and IL-6] which could induce CD4+ T cells differentiate into Th17 cells.IL-17 was detected by flow cytometry and enzyme linked immunosorbent assay (ELISA) after transfected with miR-155 mimics or inhibitor lentiviral vectors.The expression levels of miR-155, IL-17A mRNA and Ets-1 mRNA were detected using fluorescent quantitation real-time quantitative polymerase chain reaction (RT-PCR).The si-Ets and miR-155 co-function for Th17 differentiation was analyzed.Data analysis was perfoemed using one-way analysis of variance (ANOVA) test and Dunnett test for pair-wise comparison and t test.P＜0.05 was considered to be statistically significant.Results The CD4+T cells were divided into four groups (the untreated control untreat group, the treatment control treat group, the miR-155 mimnics group and miR-155 inhibitor group).IL-17 was scarcely expressed and secreted in the untreated control untreat group.The cells expression of IL-17 were significantly different among the four groups (F=160.549, P＜0.01).The cells expressing of IL-17 were higher in the miR-155 mimics group (39.86±4.62)％ than those at the miR-155 inhibitor group (22.02±2.81)％, P＜0.01) and in the treated control treat group [(19.44±1.49)％, P＜0.01].The level of IL-17 was also significantly different among the four groups (F=260.813, P＜0.01).The level of IL-17 was higher in the miR-155 mimics group [(1 509±136) pg/ml] than that in the miR-155 inhibitor group [(923± 42) pg/ml, P＜0.01);and in the treated control group [(767±94) pg/ml, P＜0.01).The expression of miR-155 (12.53±0.80 vs 1.78±0.14, 7.16±0.62, 6.47±0.92, P＜0.01) and IL-17A mRNA (46.55±6.71 vs 1.01±0.19,15.62±1.26, 14.20±2.73, P＜0.01) was significantly higher than that in the other three groups, while the expression of Ets-1 mRNA was significantly lower (0.66±0.10 vs 1.19±0.04, 1.01±0.16, 1.37±0.27, P＜0.01).si-Ets-2 was screened because it markedly inhibited the expression of Ets-1 mRNA among the three designed siRNAs.The expression of IL-17A mRNA was higher (17.19±3.58 vs 10.08±0.76, t=-3.361, P=0.028) and the expression of Ets-1 mRNA was lower (0.27±0.01 vs 0.74±0.03, t=-30.275, P＜0.01) in si-Ets-2 group than that in si-Con group when si-Ets-2 or si-Con was co-transfected with miR-155 mimics or inhibitor lentiviral vectors.The expression of Ets-1 protein was lower in si-Ets-2 group than that in si-Con group by Western blotting and the decrease was markedly obvious in the miR-155 mimics group.Conclusion miR-155 can induce CD4+T cells to differentiate into Th17 cells by inhibiting the gene expression of Ets-1.

Objective To investigate the clinical efficacy of buttress plating for patients with posterior pilon fracture.Methods From April 2012 to January 2015,12 patients with posterior pilon fracture of the distal tibia were treated in our hospital.They were 7 men and 5 women,30 to 56 years of age (average,41.2 years).According to the CT classification by Haraguchi et al.,5 cases belonged to type I,3 to type Ⅱ and 4 to type Ⅲ.All the patients underwent open reduction and internal fixation with buttress plate via either a posterolateral approach or a dual posterolateral-posteromedial approach.All the patients were available for follow-up.The clinical outcomes were evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and the visual analogue scale (VAS).The radiological evaluation was performed using the osteoarthritis-score (OA-score).Results The patients obtained an average follow-up of 21.2 months (range,from 12 to 30 months).Bone fractures united after an average of 15 weeks (range,from 13 to 19 weeks).The time for full weight walking averaged 16 weeks,ranging from 15 to 23 weeks.The ankle plantar flexion ranged from 36° to 42° (average,40.4°);the ankle dorsal extension ranged from 38° to 44° (average,42.6°).At the final follow-ups,the AOFAS scores ranged from 82 to 97 (average,88.2);the OA-score ranged from 0.6 to 0.8 (average,0.71);the VAS scores during rest,active motion and weight-bearing walking ranged from 0.5 to 0.8 (average,0.66),from 0.6 to 0.9 (average,0.82) and from 1.2 to 1.8 (average,1.41),respectively.No fracture malunion,implant loosening,pain or stiffness of the affected ankle was observed at the final follow-ups.Conclusion Buttress plating for posterior pilon fractures can lead to satisfactory clinical outcomes,because it ensures rigid fixation which in turn enables earlier postoperative mobilization.

OBJECTIVE: To investigate the inhibition effects of Tianshen Yizhi Recipe (TSYZR), a compound traditional Chinese herbal medicine, on decreased expression of nicotinic acetylcholine receptor (nAChR) and the neurotoxicity as well as lipid peroxidation induced by beta-amyloid peptide (Abeta) in human SH-SY5Y neuroblastoma cells. METHODS: The SH-SY5Y cells were treated by a certain concentration of TSYZR, and then exposed to Abeta(25-35). Methyl thiazolyl tetrazolium reduction assay was carried out to understand the influences of the drugs on cellular viability. Expressions of nAChR subunits (alpha3 and alpha7) at protein and mRNA levels were detected by Western-blotting and reverse transcription polymerase chain reaction, respectively. Lipid peroxidation was measured by thiobarbituric acid to observe the capacity of antioxidant of the drugs. RESULTS: TSYZR at a safe concentration could increase alpha7 protein in the cells, inhibit decreased expressions of alpha3 and alpha7 nAChR subunit proteins, prevent lower expression of alpha7 mRNA in SH-SY5Y cells induced by Abeta, reduce the neurotoxicity and lipid peroxidation resulting from Abeta, but had no significant effect on the lower expression of alpha3 mRNA. CONCLUSIONS: TSYZR can up-regulate the expression of alpha7 nAChR subunit protein and prevent decreased expressions of nAChRs and neurotoxicity as well as lipid peroxidation induced by Abeta. This drug may play an important therapeutic role in treatment of Alzheimer disease.

Objective To investigate the clinical application value of fluorescence polymerase chain reaction (PCR) .in the human leucocyte antigen-B27(HLA-B27) gene and gene typing detection of ankylosing spondy-litis (AS) patients .Methods A total of 43 clinical blood samples of AS and 56 samples of healthy controls were collected in Shenzhen Futian hospital for rheumatic diseases from January 2014 to March 2015 .HLA-B27 gene was detected by flow cytometry .HLA-B27 gene and gene typing was also detected by the fluorescence PCR method .Results Among 43 samples ,40 samples were HLA-B27 positive(93 .02%) by flow cytometry while 39 samples were HLA-B27 positive (90 .70%) by fluorescence PCR .The total coincidence rate was 97 .50% .Among 39 positive samples ,32 samples were HLA-B2704 positive (82 .05%) and 7 samples were HLA-B2705 positive (17 .95%) .Conclusion The fluorescence PCR is an accurate method to detect HLA-B27 gene and presents high consistency with flow cytometry .It can also detect the HLA-B27 gene typing .It may have great clinical application value and prospects .

To obtain the recombinant 18 kD protein with high purity and normal bioactivity of Cysticercus cellulosae (rCE18), E. coli cells with the rCE18 were disrupted ultra-sonically, and the inclusion bodies were washed with a solution containing 0.2% deoxycholic acid sodium (DOC)and 2% DOC, respectively. Then they were denatured with 0.9% sodium lauroyl sarcosine (SKL) followed by dialysis and gel filtration to refold and purify the target protein. At the same time, this method was compared with GST-FF affinity chromatography and recovering from SDS-PAGE gel. Biological activity of purified rCE18 was analyzed with indirect ELISA, and the purity of the products was identified using SDS-PAGE. The purity of refolded inclusion bodies exceeded 60% and the total recovery of activated protein rCE18 was about 41.3%. The specificity of rCE18 reached up to 97.2% using indirect ELISA. An effective way for purifying and refolding rCE18 expressed in E. coli as inclusion bodies was established, rCE18 with higher purity and activity was obtained, which has the potential for developing diagnosis methods of porcine cysticercosis.
Animals ; Antigens, Helminth ; biosynthesis ; genetics ; immunology ; isolation & purification ; Chromatography, Gel ; Cysticercus ; genetics ; immunology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Inclusion Bodies ; metabolism ; Protein Renaturation ; Recombinant Proteins ; genetics ; immunology ; isolation & purification