BACKGROUND:Reports have demonstrated that cytotoxicity produced by ferric oxide (Fe2O3) nanoparticles is associated with cellular lipid peroxidation. Whether Fe2O3 nanoparticles have toxicity to macrophages, and what is the association of toxic mechanism and oxidization?OBJECTIVE: To observe the effects of different concentrations of Fe2O3 nanoparticles on the oxidative damage of macrophages.DESIGN: A controlled observation experiment.SETTING: School of Public Health, Southeast University.MATERIALS: RAW264.7 cells were peritoneal macrophages of mouse and purchased from Shanghai Institute of cells, Chinese Academy of Sciences. Fe2O3 nanoparticles (30 nm) suspension was provided by Department of Biomedical Engineering, Southeast University). Fe2O3 nanoparticle suspension was placed in 60 ℃ water for 10 hours,then in 37 ℃ water overnight. This procedure was repeated 3 times for germicidal treatment. Then, the suspension was packed into small bottles and stored at 4 ℃ for later use. DMEM high glucose culture fluid (Gibco Company,USA); trypsinase (Difco Company, USA, imported); new-bom calf serum(Sijiqing Company, Hangzhou); hydrogen dioxide (H2O2, Gibco Company); Kits for measuring hydrogen dioxide(H2O2), hydroxy radical (·OH), superoxide anion radical (O2·-), lactic acid dehydrogenase, ultramicro ATP enzyme and Coomassie brilliant blue protein levels (Jiancheng Biotechnique Co., Ltd.,Nanjing).METHODS: This experiment was carried out in the laboratory of Department of Labor and Environmental Health, School of Public Health, Dongnan University between March 2006 and July 2006. RAW264.7 cells (Abelson murine leukemia virus-induced tumor) were cultured in DMEM (Gibco Company) containing 100 g/L fetal bovineserum, 100 000 U/L penicillin and 100 mg/L streptomycin in the environment of 5% CO2. Cell growth was observed under an inverted radical in the cells: 1.5×108 L-1 macrophages were inoculated to 24-well plate, 1 mLa well. After the macrophages were cultured for 24 hours in incubation at 37 ℃ in a humidified atmosphere containing 5% CO2. 1.070 0, 0.5350 and 0.2675 g/L Fe2O3 nanoparticles (30 nm) suspension-intervened macrophages were set as Fe2O3 nanoparticle group, and normal saline group was set as control group. Following the intervention of nanoparticles, macrophages were disrupted with Determination of the activities of lactate dehydrogenase (LDH), Na+-K+-ATPase and Ca2+-Mg2+-ATPase: Macrophages in the Fe2O3 nanoparticle group and control group were treated as above. The activities of LDH in culture medium were determined according to the instruction of reagent kit (Nanjing Jiancheng Bioengineering Co., Ltd). And the activities of Na+-K+-ATPase and Ca+-Mg2+-ATPase were also determined according to the instruction of reagent kit (Nanjing Jiancheng Bioengineering Co., Ltd) at low temperature. MAIN OUTCOME MEASURES: ①Effects of different concentrations of Fe2O3 nanoparticles on the production of H2O2, ·OH and O2·- in RAW264.7 cells.②Effects of different concentrations of Fe2O3 nanoparticles on the activities of LDH ,Na+-K+-ATPase and Ca2+-Mg2+-ATPase in RAW264.7 cell culture fluid.RESULTS: ① Level of ·OH free radical in Fe2O3 nanoparticle 0.267 5, 0.535 0, 1.070 0 g/L groups was higher than that in control group, respectively [(0.605±0.066), (0.410±0.080), (0.764±0.051), (0.285±0.057)mkat/g, P ＜ 0.05]; Level of respectively [(9.935±1.159), (8.912±0.131), (13.479±0.752), (5.635±0.475)μkat/g,P ＜ 0.05]; Level of H2O2 in Fe2O3 nanoparticle 1.070 0 g/L group was higher than that in the control group [(14.695±2.815), (2.397±0.399) mmol/L, P ＜increased (P ＜ 0.05). Fe2O3 nanoparticles had effects on the activities of Na+,K+-ATPase and Ca2+,Mg2+-ATPase. With the increase of dose of Fe2O3 nanoparticles, the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase were gradually decreased. There were significant differences as compared with control group (P ＜ 0.05)CONCLUSION:Increasing dose of Fe2O3 nanoparticles wouldcause more H2O2,·OH and O2·- free radicals in the cells, increase cell membrane permeability and inhibit the activities of LDH, Na+-K+-ATPase and Ca2+-Mg2+-ATPase.

Objective:To study the protective effect of neoeriocitrin on PC12 cell injury induced by amyloid β protein fragment 25-35 (Aβ 25-35), and explore its role in preventing and treating AD. Methods:The MTT method was used to detect the effect of Neoeriocitrin on the proliferation of normal PC12 cells and Aβ 25-35 damaged PC12 cells, and the intervention concentration of neoeriocitrin solution was screened. The PC12 cells were divided into the normal control group, model group, estradiol group and neoeriocitrin group according to the random number table method. The normal control group and model group were cultured with DMEM for 24 h; DMEM and 1×10 -3 μmol/L estradiol solution were added to the estradiol group; DMEM and 6×10 4 μmol/L neoeriocitrin solution were added to the neoeriocitrin group. After 2 hours of culture, except for the normal control group, the remaining groups were added with 20 μmol/L Aβ 25-35 stock solution, and the culture was continued for 22 h. AnnexinV-FITC/PI double labeling was used to detect apoptosis rate, Western blot method was used to detect estrogen receptor β (estrogen receptor β, ERβ) and p-P38/P38 protein expression. The content of acetylcholine (ACh) was detected, and the activity of Choline acetyl transferase (ChAT) and Acetylcholin esterase (AChE) in the supernatant of PC12 cells were detected. Results:Compared with the model group, the cell apoptosis rate [(8.080 ± 0.578)% vs. (18.500 ± 0.870)%] significantly decreased ( P<0.01), the expression of ERβ protein (0.348 ± 0.042 vs. 0.273 ± 0.006) significantly increased ( P<0.01), p-P38/P38 protein expression (0.372 ± 0.058 vs. 0.571 ± 0.063) significantly decreased ( P<0.01), the content of ACh [(14.319 ± 1.039) μg/mg vs. (9.157 ± 1.605) μg/mg], ChAT activity [(0.715 ± 0.053) U/mg vs. (0.280 ± 0.093) U/mg] significantly increased ( P<0.01), AChE activity [(2.607 ± 2.048) U/mg vs. (6.038 ± 1.867) U/mg] significantly reduced ( P<0.01). Conclusions:Neoeriocitrin can promote the proliferation of PC12 cells damaged by Aβ25-35, inhibit cell apoptosis, and has a certain cytoprotective effect. Its mechanism may be related to the regulation of ERβ expression and inhibition of P38 protein phosphorylation, thereby improving the function of the cholinergic system related.

OBJECTIVETo screen phosphopeptide specific for acute leukemia.

METHODSMononuclear cells from bone marrow were collected from 16 newly diagnosed acute lymphoblastic leukemia (ALL) and 20 acute myeloid leukemia (AML) patients. Peptides were extracted and purified, analyzed by immunoprecipitation and liquid chromatography coupled with tandem mass spectrometry (LC-MS).

RESULTS(1) Non-receptor tyrosine kinase family members Fyn, Yes, Src widely expressed in acute leukemia; (2) Some phosphopeptides, including non-receptor tyrosine kinase family members Abl/iso1 and Abl, non-receptor Ser/Thr protein kinase family members Bcr, JNK2, JNK2 iso2, Adaptor/scaffold members Cas-L, Cbl, CrkL CENTD1 (Centaurin delta1) ZO2, transcriptor GFR-1 and phosphatase SHIP-2 were detected in Ph positive ALL, but not in other kinds of ALL. (3) Hck, Lyn and Fgr selectively expressed in AML (except AML-M(3)).

CONCLUSIONSome phosphopeptides were specific for ALL and AML, and may be useful for diagnosis and therapy of acute leukemia.

Chromatography, High Pressure Liquid ; Humans ; Immunoprecipitation ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Neoplasm Proteins ; analysis ; Phosphopeptides ; analysis ; Phosphorylation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Proteomics ; Tandem Mass Spectrometry

Objective To investigate the value of three-dimensional speckle tracking imaging (3D-STI) in assessment of left ventricular (LV) strains. Methods Thirty healthy young adults examined by two-dimensional speckle tracking imaging (2D-STI) and 3D-STI. And the results of LV measurements were compared, which included mean peak systolic longitudinal strains, radial strains and circumferential strains. Also, the time consumption of these two methods was compared. Results The time needed for 3D-STI in acquisition and analysis of the images were (309.3±23.4)s, (305.5±11.2)s, while the time for 2D-STI were (490.6±14.4)s, (1261.4±39.9)s. The differences were signiifcant(t=-21.81, 69.94, both P<0.01). The global mean peak systolic radial strains was (48.59±7.68)%by 3D-STI and (33.25±7.27)%by 2D-STI. The difference was signiifcant(t=9.16, P<0.01). The global mean peak systolic longitudinal and circumferential strains were (-17.66±3.14)%, (-17.13±2.29)% by 3D-STI and (-21.35±2.46)%, (-21.97±3.84)% by 2D-STI. The differences were signiifcant(t=5.33, 5.99, both P < 0.01). The 3D-STI strains were different at different levels of LV. The longitudinal, circumferential and radial 3D-STI strains were largest at middle levels. However, 2D-STI strains didn′ t show such trend. Peak strains measured by 3D-STI and 2D-STI showed high inter-observer and intra-observer agreement in Bland-Altman chart. Conclusion 3D-STI is a novel, convenient and reproducible method to evaluate the strains of LV.

AIM:To analyze the association of intrauterine growth retardation (IUGR) and retinopathy of prematurity (ROP).METHODS:A retrospective analysis of a case series included in ROP screening from January 2011to December 2015 was performed in Suzhou Municipal Hospital.Totally 2527 children (5054 eyes) underwent screening.According to the gestational age,the data was divided into 4 groups (≤32wk,＞32 and ≤34wk,＞34 and ≤37wk,＞37wk).Every group was divided into two groups (IUGR group and no IUGR group) respectively.We compared the incidence of ROP in IUGR and non IUGR group.RESULTS:Of all the 2527 children,IUGR group were 702 including 78 ROP children,and non IUGR group were 1825 including 329 ROP children.There were 991 children were divided into ≤ 32wk group,including 63 IUGR in which 27 children were screened out ROP(42.9％) and 928 non IUGR in which 274 children were screened out ROP (29.5％),the difference on the incidence of ROP was statistically significant (X2 =4.958,P=0.026).There were 1025 children were divided into ＞ 32 and ≤ 34wk group,including 232 IUGR in which 33 children were screened out ROP(14.2％) and 793 non IUGR in which 51 children were screened out ROP (6.4％) and the difference was statistically significant (x2 =14.488,P＜0.001).There were 464 children were divided into ＞ 34 and ≤ 37wk group,including 374 IUGR in which 18 children were screened out ROP(4.8％) and 90 non IUGR in which 4 children were screened out ROP (4.4％) and the difference was not statistically significant (Fischer exact test,P=1).There were 47 children were divided into ＞37wk group,including 33 IUGR and 14 non IUGR,none were screened out in the two groups.CONCLUSION:Intrauterine growth retardation was closely related to the incidence of ROP.In the preterm infants with gestational age less than 34wk,the incidence of ROP in children with intrauterine growth retardation is significantly higher than that in children without intrauterine growth retardation.

·AIM: To retrospectively analyze the prevalence of retinopathy of prematurity(ROP) in 3471 neonates in Suzhou Municipal Hospital. ·METHODS: A total of 3471 children (1947 males, 1524 females) were screened for ROP in Suzhou Municipal Hospital from January 2010 to September 2016 using binocular ophthalmoscope or ( and) RetCamⅡ. First examination was performed from 4-6wk after birth. The ocular findings were recorded according to the International Classification of ROP and The Early Treatment for ROP. Only the more aggressive eye of bilateral asymmetrical cases was counted for statistical purpose. Children with ROP in both binocular or single eye were counted in 1 case, and the cases required surgeries were defined as severe cases. The prevalence of ROP and severe ROP in recent 6a were analyzed retrospectively. ·RESULTS: The overall relevance ratio of ROP and severe ROP was 17.03% and 1.15%. The relevance ratio of ROP and severe ROP of the males were 16.38% and 1.08%,and of the females were 17.85% and 1.25%, the results were not statistically different (x2= 1. 296, P =0.255). The relevance ratio of ROP and severe ROP of the single birth infants were 17.61% and 1.13%, and of the multiple birth infants were 15.13% and 1.23%,the results were not statistically different (x2=2.706, P=0.100). The children were divided into 5 groups according to the birth weight. The relevance ratio of ROP with birth weight<1000g,1000-1499g,1500-1999g,2000-2499g and ≥2500g were 75. 00%, 36. 17%, 10. 75%, 6. 86% and 3. 77%respectively with significant differences (There were significant differences between the three groups which the birth weight <2000g, P<0.005). The relevance ratio of severe ROP were 36.54%, 1.68%, 0.31%, 0.19% and 0 respectively in these birth weight groups (There were significant differences between the three groups which the birth weight <2000g,P<0.005). The children were divided into 4 groups according to gestational weeks, the relevance ratio of severe ROP of gestational age<28wk,28-31wk, 32-36wk and ≥37wk were 69. 12%, 29. 91%, 8.28% and 3.33% respectively with significant differences (There were significant differences between the three groups which the gestational age <37wk, P<0.005). The relevance ratio of severe ROP were 25%, 1.52%, 0.24% and 0 in these gestational age groups respectively (There were significant differences between the three groups which the gestational age <37wk,P<0.005). · CONCLUSION: The detection rate of ROP in 3471 premature infants was 17. 03%, the severe ROP was 1.15%. There was no evidence that sex and birth were related to ROP, but lower birth weight and smaller gestational age increased the detection rate of ROP.

AIMTo study the effect of the recombined human growth hormone(rhGH) on secretory immunoglobulin A (sIgA), epidermal growth factor (EGF) in rats with obstructive jaundice.

METHODSSixty Wistar rats were randomly divided into four groups, sham-operated (group A), common biliary duct-ligated (group B), biliary duct-ligated plus rhGH-treat for one week (group C), biliary duct-ligated plus rhGH-treat for two weeks (group D), each group had 15 rats. Except group A, the rats of other groups were operated with biliary duct-ligated. Until two weeks after operation, the rats of group A and B were killed. After operation, the rats of group C were treated with rhGH hypodermic injection (0.75 U x kg(-1) x d(-1)) for one week, and then killed. The rats of D group were treated with rhGH hypodermic injection (0.75 U x kg(-1) x d(-1)) for two weeks, and then killed. All procedures were performed aseptically. Total bilirubin (TB), alkaline phosphatase (ALP), prealbumin(PA), insulin-like growth factor (IGF-1), sIgA, EGF were measured.

RESULTSCompared with group A, in group B, C, D, serum level of PA, IGF-1 and sIgA, EGF level of gastric and intestinal juice were lower, but TB, ALP were higher, there were significant difference. Compared with group B, the rats with treatment of rhGH in group C and D had higher sIgA and EGF and lower intestinal bacterial translocation.

CONCLUSIONIn objective jaundice rats, rhGH can protect their hepatic function, intestinal physical-barrier function and immune-barrier function, and reduce intestinal bacterial translocation.

Animals ; Bacterial Translocation ; Epidermal Growth Factor ; metabolism ; Growth Hormone ; pharmacology ; Humans ; Immunoglobulin A, Secretory ; metabolism ; Jaundice, Obstructive ; metabolism ; Rats ; Rats, Wistar ; Recombinant Proteins ; pharmacology

Objective :To construct and identify a ScFv phage display library against human umbilical cord mesenchymal stem cells.Methods: BALB/c mice were immunized with cultured UC-MSCs.After the third immunization,the total RNA was extracted from the spleen cells of the immunized BALB/c mice and purified by affinity chromatography with mRNA Purification Kit.The heavy-chain and light-chain variable region genes(VH and VL) were amplified by PCR using relevant primers.PCR products of VH and VL genes were cloned into the phagemid vector pSEX81 and electroporated into the XL1-Blue strain of E.coli.The ScFv phage display library against human umbilical cord mesenchymal stem cells was constructed and the capacity of library was measured.The library was panned by three cycles and screened with purified UC-MSCs.The percentage of clones containing a full-length scFv-encoding insert and their diversity was determined for unselected and selected libraries.Results: The amplified fragments of VH and VL genes by RT-PCR were about 399bp and 357bp,respectively.VH and VL genes were all successfully cloned into the phagemid vector pSEX81,which were confirmed by the amplication of 786bp full-length scFv fragments by PCR.The ScFv phage display library had a capacity of approximately 2?107 cfu.After three cycles of panning,PCR of plasmid DNA prepared from 15 individual phage clones showed that the recombination rate increased from 93% to 100%.BstN1 fingerprinting of insert DNA showed that the diversity of clones decreased with increasing rounds of selection.After three rounds of selection,3 clones showed an identical restriction enzyme pattern.There was a 330-fold enrichment of library phage after 2 rounds of selection and after 3 rounds,a further 8-fold enrichment of library phage was obtained.Conclusion: The ScFv phage display library against human umbilical cord mesenchymal stem cells was successfully constructed.It can be used for succeeding screening of specific antibody against human umbilical cord mesenchymal stem cells and further studying of the cell surface molecules of mesenchymal stem cells.

Objective To discuss the optimal clinical diagnosis and treatment of ectopic ACTH syndrome with occult tumors. Methods Clinical features, imaging examinations and treatment of 17 patients with ectopic ACTH syndrome were described and compared. Results All patients illustrated the typical clinical features of Cushing’s syndrome. They had hypokalemic alkalosis, elevated serum cortisol and plasma ACTH levels. In the high-dose dexamethasone suppression tests, most patients failed to suppress serum cortisol and 24-hour urinary cortisol. CT and MRI are useful imaging modalities to localize the ACTH-secreting tumor in patients with ectopic ACTH syndrome. The patients with overt ACTH-secreting tumors had surgical curative resection soon after diagnosis. Among patients with occult ACTH-secreting tumors, three underwent subtotal bilateral adrenalectomy, two underwent right adrenalectomy, four received inhibitor of steroidogenesis aminoglutethimide. Their hypercortisolism was controlled. Conclusion Surgical curative resection is the optimal treatment of ectopic ACTH syndrome with overt ACTH-secreting tumor. Bilateral adrenalectomy, right adrenal ectomy or chemotherapy to control hypercortisolism is an available treatment of ectopic ACTH syndrome with occult ACTH-secreting tumors.

Inherited afibrinogenemia is a rare autosomal recessive bleeding disease characterized by complete absence of fibrinogen in blood. To identify the genotype in a Chinese family with inherited afibrinogenemia, the samples of peripheral blood were collected from 6 members of 3 generations. The activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) and fibrinogen (Fg, clauss) were tested. Fg was also analyzed by using immunoturbidimetry method. DNAs of six members were extracted by using a DNA extract kit. All the exons and exon-intron boundaries of the three fibrinogen genes were amplified by using PCR and analyzed by direct sequencing. The results showed that the parents of proband were 3 degree consanguinity. A homozygous c.934_935insA in FGA was found in proband which results in the change of protein p.Ser312fsX42. The parents, grandmother, maternal grandmother and father's sister were all detected with heterozygous mutation which was same as that in proband. In conclusion homozygous c.934_935insA in FGA is a cause of inherited afibrinogenemia and a novel mutation being reported.
Afibrinogenemia ; etiology ; genetics ; Child ; Exons ; Female ; Fibrinogen ; genetics ; Frameshift Mutation ; Heterozygote ; Humans ; Male ; Pedigree