Objective: To observe the effect of electroacupuncture (EA) pretreatment on the protein expression of c-fos in fastigial nucleus (FN) and lateral hypothalamus area (LHA) in rats with acute myocardial ischemia-reperfusion injury (MIRI), and to explore the role and mechanism of FN and LHA in EA at the Heart Meridian fighting against acute MIRI reaction. Methods: Seventy Sprague-Dawley rats were randomly divided into a sham operation group, a model group, an EA-Heart Meridian group and an EA-Lung Meridian group, with 14 rats in each group; an LHA lesion plus EA-Heart Meridian group (LHA+EA-Heart Meridian group) and a FN lesion plus EA-Heart Meridian group (FN+EA-Heart Meridian group), with 7 rats in each group. Except the sham operation group, the left anterior descending branch of coronary artery was ligated to establish acute MIRI rat models in the other 5 groups. In the three groups with EA-Heart Meridian treatment, Shenmen (HT 7) and Tongli (HT 5) were selected; Taiyuan (LU 9) and Lieque (LU 7) were selected in the EA-Lung Meridian group. All the EA groups received EA stimulation prior to modeling, with 1 mA in current intensity and 2 Hz in frequency, 20 min each time, once a day for a total of 7 d. The sham operation group and the model group did not receive EA stimulation. The electrocardiogram was observed in the rats to analyze the ST-segment deviation and cardiac arrhythmia score. The expression of c-fos protein in FN and LHA was detected by immunohistochemistry method. Results: Compared with the sham operation group, the ST-segment deviation, cardiac arrhythmia score and the expression of c-fos protein in the FN and LHA increased significantly in the model group (all P<0.05). Compared with the model group, the ST-segment deviation, cardiac arrhythmia score and the expression of c-fos protein in FN and LHA decreased significantly in the EA-Heart Meridian group (all P<0.05). Compared with the EA-Heart Meridian group, the ST-segment deviation and cardiac arrhythmia score increased significantly in the EA-Lung Meridian group, LHA+EA-Heart Meridian group and FN+EA-Heart Meridian group (all P<0.05); the expression of c-fos in FN increased significantly in the EA-Lung Meridian group and LHA+EA-Heart Meridian group (both P<0.05); the expression of c-fos in LHA increased significantly in the EA-Lung Meridian group and FN+EA-Heart Meridian group (both P<0.05). Conclusion: FN and LHA are involved in the mechanism of EA at Heart Meridian to improve the acute MIRI reactions, and the cerebellum may participate in the improvement of cardiac function by EA through the cerebellum-hypothalamus projection.

ObjectiveTo investigate the iodine nutritional status of pregnant women,newborn heel blood thyroid stimulating hormone(TSH) level and their relationship with urinary iodine(UI) level during pregnancy in Zhoupu and Kangqiao districts of Pudong New Area of Shanghai.Methods A total of 993 urinary samples(the first,second and third trimesters of pregnancy were 200 people,respectively),breast feeding(193 people) and non-pregnant women (200 people) in Zhoupu and Kangqiao districts of Pudong New area were collected from Apr 2009 to Dec 2010.Two hundred copies of neonatal heel blood samples were collected.Median of UI was measured by arsenic-cerium catalysis.TSH in neonatal heel blood was analyzed 72 h after birth by time resolved fluoroisnmunoassay(TRFIA).ResultsMedian UI of all pregnant women was 161.35 μg/L,and that in third trimesters of pregnancy( 126.35 μg/L) was lower than that of the first,the second,the breast feeding and non-pregnant women (178.80,180.50,167.90,163.40 μg/L,all P＜ 0.05).The percentage of UI level less than 150 μg/L in the third trimester[57.5％(115/200) ] was higher than that of the first[39.0％(78/200) ],the second[39.5％(79/200) ],the breast feeding [ 16.6％ (32/193) ] and non-pregnant women [ 23.0％ (46/200) ],respectively (all P ＜ 0.05).The percentage of UI level higher than 300 μg/L in the first [9.0％(18/200)],the second[8.0％(16/200) ] and the third trimester [ 5.0％ ( 10/200 ) ] of pregnancy was lower than that of the breast feeding [ 20.2％ (39/193) ] and nonpregnant [20.5％(41/200) ] women,respectively(all P ＜ 0.05).The level of neonatal heel blood TSH was(2.92 ± 1.83)mU/L,the range was 0.01 - 9.76 mU/L,11.0％(22/200) of the neonates heel blood TSH level(5 mU/L)exceeded the ratio of World Health Organization (WHO) standard ( ＜ 3％ ) suitable for iodine nutrition.Conclusions The overall level of iodine nutrition among pregnant women in Zhoupu and Kangqiao districts of Pudong New Area of Shanghai is in the appropriate range,but the pregnant women in the third trimester is in mild iodine deficiencies,and the neonates in these districts may be prone to iodine deficiency.Monitoring of iodine nutrition of pregnant women should be strengthened and iodine supplementation should be done scientifically.

Objective To evaluate the diagnostic value of urine Cystatin C(Cys C) for renal function impairment in neonates with hypoxic-ischemic encephalopathy(HIE).Methods The urine Cys C concentration was measured by enzyme linked immunosorbent assay(ELISA) in 47 cases of HIE newborns(25 cases were mild HIE and 22 cases were moderate-severe HIE) within 3 days after their birth.Twenty-three cases without perinatal asphyxia or other factors which could result in renal function impairment were selected as control group.Urine Cys C with urine retinal-bindingprotein(RBP),?2-microglobulin(?2-MG) and fractional sodium excretion(FENa%) were analyzed by kolmogorov-smirno in each group.Results Compared with control group,the concentration of urine Cys C,RBP and the levels of FENa% in HIE newborns were significantly elevated.The levels of urine Cys C in moderate-severe HIE newborns were significantly higher than those in mild HIE newborns(Pa

OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

Objective To investgate prevalence and clinical significance of antiproteinase 3(PR3)an- tibodies in Wegener's granulomatosis(WG)and other vasculitis patients.Methods One hundred and eleven systemic vasculitis patients with WG(9 cases,including 21 serums of tracking WG patients)and other systemic vasculitis(102 cases),403 secondnary vasculitis CTD(SLE 213 cases,RA 135 cases),nephritis 62 cases,30 healthy subjects were examined for anti-PR3 and anti-MPO antibody by enzyme-linked immunosorbent assay (ELISA)and ANCA by indirect immunofluorescence(IIF)was performed.Result Anti-PR3 positive were 23 in 588 serums of patients.The prevalence of anti-PR3 positive was WG(16/21,71.4 %),other systemic vas- culitis were not found anti-PR3,SLE(6/213,2.8%),RA(1/135,0.7%).In particular,the prevalence of anti- PR3 and cANCA with WG tended to be higher in the patients with other systemic and secondnary vasculitis (P＜0.05).The sensitivity and specificity of anti-PR3 for diagnosis of WG were 71.42% and 98.58%.The sensi- tivity and specificity of combination anti-PR3 and cANCA were 61.90% and 99.82%.Anti-PR3 and cANCA are associated with treament of WG.Conclusion Anti-PR3 antibody has high specificity for diagnosis of RA. Detection of anti-PR3 and cANCA at the same time can improve the specificity considerably.As sensitive markers of WG,anti-PR3 antibody may be useful for diagonosis and early treament.Anti-PR3 also may be useful for activity and relapse of WG.

Objective: To construct, express and purify ScFvl4/EGFP fusion proteins which containing Arg9, and to study their binding activities and internalization functions. Methods: Arg9 gene was recombined into 5' terminal, 3' terminal of ScFv/EGFP gene and between them respectively before they were cloned into the expression vector pET32a. After induced in E. coli BL21 and purified, their binding activities and internalization were respectively analyzed by indirect ELISA and indirect immunofluorescence analysis. Results: DNA sequencing and restriction endonuclease digestion proved that the four fusion genes were correctly constructed. SDS-PAGE analysis and Western blot showed that they were successfully expressed and purified. Indirect ELISA confirmed that the expressed products had antigen specific binding activities. Indirect immunofluorescence analysis revealed the fusion protein containing Arg9 at its N terminal had much better internalization function, but never internalized into the cells which do not express HBsAg. Conclusion: The four fusion genes were constructed, expressed and purified successfully. The purified fusion proteins maintained the binding activities to HBsAg and the fusion protein containing Arg9 at its N terminal had much better internalization effect.

Objective To understand the risk situation of imported schistosomiasis and its main influencing factors in Shang-hai City from 2005 to 2015. Methods A retrospective survey was performed,and the database was established based on the da-ta collection of imported schistosomiasis,Oncomelania hupensis snail situation and mobile population in Shanghai City from 2005 to 2015. Results From 2005 to 2015,the mobile population in Shanghai City increased by 123.92％,which reached about 9816500,and 54.70％of them were located in the inner suburban districts. The accumulated areas with snails of 7.13 hm2 were found in 16 towns of 4 outer suburbs(excluding Chongming District). A total of 23 cases of imported schistosomiasis were found in Shanghai City,and the number of the imported schistosomiasis cases was on the downward trend(rs=-0.782,P=0.004). From 2005 to 2009,11 out of 13 imported schistosomiasis cases(84.62％)were distributed in the inner suburban dis-tricts where no O. hupensis snails were found at the same time. From 2010 to 2015,9 out of 10 imported schistosomiasis cases (90％)were distributed in the outer suburban districts where O. hupensis snails were found at the same time. Conclusion The number of imported schistosomiasis cases in Shanghai is on the downward trend. However,the threat of imported schistosomiasis to the prevention and control of schistosomiasis in Shanghai City should not be ignored.

The aim of this study was to investigate the homing characteristics of bone marrow cells in leukemia mice after allogenic bone marrow transplantation with different conditioning regimens on the basis of a leukemia mouse model. Allogenic bone marrow transplantation was performed after three different kinds of conditioning regimen, including nonmyeloablative conditioning regimen (5 Gy (60)Co γ ray total body irradiation, A group), radiotherapeutic myeloablative conditioning regimen (9 Gy (60)Co γ ray total body irradiation, B group) and chemotherapeutic myeloablative conditioning regimen (large dose chemotherapy, C group). In the recipient mice, the nucleated cell number in peripheral blood, bone marrow and spleen was counted, the percentage of positive cells capable of connecting with FITC labeled anti-mouse H-2K(b) antibody was detected by flow cytometry and the homing ratio in bone marrow and spleen was calculated at 24, 48, 72, 96 h after bone marrow transplantation. The results showed that donor myeloid cells displayed homing and then mobilization (going out of home) in group A; homing, mobilization, and rehoming in group B and C, and there was a little delay of homing in the spleen in group C. In bone marrow, the homing efficiency of A group was the highest in early period and the lowest [(0.90 ± 0.09)%] in the fourth day with the mobilization of myeloid cells (P < 0.05), and the homing efficiency of B and C groups was lower in the early period and the highest [(2.17 ± 0.26)%, B group] in the fourth day with the rehoming of myeloid cells (P < 0.05). In spleen, the homing efficiency was similar to that in bone marrow and there still was a little delay in C group. It is concluded that the homing ratio is high in the early period and decrease obviously in 72 h after bone marrow of leukemia mice treated with nonmyeloablative conditioning regimen. The homing ratio is low in the early period and increases obviously in 72 h after bone marrow of leukemia mice treated with radio-or chemotherapeutic myeloablative conditioning regimens. The homing ratio does not obviously change between the early period and 72 h after bone marrow of leukemia mice treated with chemotherapeutic myeloablative conditioning regimen, and lies between group A and B.
Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; methods ; Cell Count ; Female ; Graft Survival ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Spleen ; cytology ; Transplantation Conditioning ; methods ; Whole-Body Irradiation

OBJECTIVETo determine the potential urinary biomarkers of metabolic syndrome (MS) with early renal injury and establish diagnostic models by magnetic bead-based separation and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

METHODSParticipants were selected from the epidemiologic study on MS and renal involvement among residents in Pinggu district, Beijing. Eight-hour overnight urine samples were fractionated by means of magnetic bead-based weak cation exchange chromatography and subsequently analyzed with MALDI-TOF-MS. Wilcoxon test and random forests were used to screen differential protein peaks of MS patients with early renal injury, then combined with genetic algorithm and support vector machine, respectively, to establish diagnostic models.

RESULTSTotally 54 cases of MS without renal injury and 46 cases of MS with early renal injury were enrolled. Totally twenty protein peaks were up-regulated in the urine of MS patients with early renal injury by Wilcoxon test (P < 0.05); random forests algorithm revealed twelve protein peaks up-regulated in the urine of MS patients with early renal injury (importance value of mean decrease in accuracy > 0.005). Genetic algorithm based model showed 82.6% sensitivity, 84.3% specificity, and 83.5% accuracy by a 10-fold cross-validation in identifying MS patients with early renal injury; correspondingly, the support vector machine based model reported 89.2% sensitivity, 81.1% specificity and 85.5% accuracy. Four protein peaks were included in two diagnostic models with mass-to-charge ratios of 2756.98, 3019.11, 9077.04, and 10 054.26.

CONCLUSIONSThe urinary proteome patterns of MS with early renal injury were successfully established with magnetic bead-based separation and MALDI-TOF-MS technology. A series of urinary differential expressing protein peaks were identified with bioinformatics tools. Diagnostic models combining cluster of protein peaks are capable of differentiating MS patients with early renal injury from those without renal injury. The different urine protein excretion patterns revealed in this study provide urinary candidate biomarkers of MS patients with early renal injury for future identification and biological roles investigation.

Adult ; Biomarkers ; urine ; Chromatography, Ion Exchange ; methods ; Female ; Humans ; Kidney Diseases ; etiology ; urine ; Male ; Metabolic Syndrome ; complications ; urine ; Middle Aged ; Proteome ; analysis ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Urine ; chemistry

OBJECTIVETo identify the fibrinogen (Fg) gene mutations in a Chinese pedigree of congenital afibrinogenemia.

METHODSThe plasma Fg activity and protein of the proband and his family members were detected. Genomic DNA was isolated from the peripheral blood mononuclear cells. All the exons and exon-intron boundaries of fibrinogen gene were amplified by PCR and sequenced thereafter.

RESULTSTwo mutations, 7972 del G in FGB and T2543A in FGG, were found in the proband.

CONCLUSIONSFGG2543 is a polymorphism site, which lead to the polymorphism of gamma144 I/K. The G deletion at base 7972 of FGB contributes to the frameshift mutation after amino acid 419, resulting in the truncated beta chain without the terminal 27 amino acids. The latter may contributes to the pathogenetic mechanisms in Chinese congenital afibrinogenemia patients. The G deletion at base 7972 of FGB is identified for the first time.

Adult ; Afibrinogenemia ; congenital ; genetics ; metabolism ; Base Sequence ; Blotting, Western ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Fibrinogen ; genetics ; Humans ; Introns ; genetics ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction