Objective By detecting the variation of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) DNA methylation in preeclampsia-like mouse models generated by different ways, to explore the roles of multifactor and multiple pathways in preeclampsia pathogenesis on molecular basis. Methods Established preeclampsia-like mouse models in different ways and divided into groups as follows: (1) Nw-nitro-L-arginine-methyl ester (L-NAME) group: wild-type pregnant mouse received subcutaneous injection of L-NAME;(2) lipopolysaccharide (LPS) group:wild-type pregnant mouse received intraperitoneal injection of LPS; (3) apolipoprotein C-Ⅲ (ApoC3) group: ApoC3 transgenic pregnant mouse with dysregulated lipid metabolism received subcutaneous injection of L-NAME;(4)β2 glycoprotein I (β-2GPI) group:wild-type pregnant mouse received subcutaneous injection ofβ-2GPI. According to the first injection time (on day 3, 11, 16 respectively), the L-NAME, LPS and ApoC3 groups were further subdivided into:pre-implantation (PI) experimental stage, early gestation (EG) experimental stage, and late gestation (LG) experimental stage.β-2GPI group was only injected before implantation. LCHAD gene methylation levels in placental were detected in different experimental stage. Normal saline control groups were set within wild-type and ApoC3 transgenic pregnant mice simultaneously. Results (1) CG sites in LCHAD DNA:45 CG sites were detected in the range of 728 bp before LCHAD gene transcription start site, the 5, 12, 13, 14, 15, 16, 19, 24, 25, 27, 28, 29, 30, 31, 32, 34, 35, 43 CG sites were complex sites which contained two or more CG sequences, others were single site which contained one CG sequence. The 3, 5, 6, 11, 13, 14, 18, 28 sites in L-NAME, LPS, ApoC3 and β-2GPI groups showed different high levels of methylation; the 16, 25, 31, 42, 44 sites showed different low levels of methylation; other 32 sites were unmethylated. (2) Comparison of LCHAD gene methylation between different groups:the methylation levels of LCAHD gene at 3, 11, 13, 14, 18 sites in L-NAME, LPS, ApoC3 andβ-2GPI groups were significantly higher than those in the normal saline control group (P<0.05); and the methylation levels of 42, 44 sites in these groups were significantly lower than those in the normal saline control group (P<0.05). (3) Methylation of LCHAD gene at the same site between different experimental stages: ① The 3, 11, 18 sites of EG experimental stage was significantly lower than PI and LG experimental stage in L-NAME group (P<0.05);the 3, 11, 18 sites of PI experimental stage was significantly lower than EG and LG experimental stage in LPS group (P<0.05);these sites of PI experimental stage was significantly higher than EG and LG experimental stages in ApoC3 group (P<0.05).②The methylation of site 5 in L-NAME and LPS groups were significantly higher than that of the normal saline control group (P<0.05), and the LG experimental stages were significantly higher than other stages, but in ApoC3 group , only PI and EG stages were significantly higher than the normal saline control group (P<0.05).③At site 6 in L-NAME group which showed high methylation level was significantly higher than the same site in other groups which showed low methylation level (P<0.05).④At 13, 14 sites, earlier preeclampsia onset caused a lower methylation level in L-NAME group, but PI experimental stage was significantly higher than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05). ⑤ At site 28, earlier preeclampsia onset caused a higher methylation level in L-NAME group, but PI experimental stage was significantly lower than EG and LG experimental stages in LPS group (P<0.05), EG experimental stage was significantly higher than PI and LG experimental stages in ApoC3 group (P<0.05).⑥The 16, 25, 31 sites in ApoC3 group were significantly higher than other groups (P<0.05). ⑦ At site 42 in β-2GPI group was unmethylated, but it in other groups showed low methylation level, the methylation level of site 42 inβ-2GPI group was significantly lower than that in other groups (P<0.05). Conclusions The methylation of 6 and 42 CG sites may be related to LCHAD gene expression in placenta of L-NAME and β-2GPI induced preeclampsia-like models respectively;LCHAD gene expression and DNA methylation may not have obviouscorrelation in LPS and ApoC3 induced preeclampsia-like models. Differences exist in LCHAD DNA methylation in preeclampsia-like models generated by different ways, revealed a molecular basis to expand our understanding of the multi-factorial pathogenesis of preeclampsia.

Objective:To evaluate the efficiency of using artificial intelligence reading label system in diabetic retinopathy (DR) grading training among junior ophthalmologists and medical students.Methods:520 diabetic fundus images were randomly divided into 8 groups with 65 images in each group. 13 junior ophthalmologists and medical students were selected as the research objects. Each of them read 8 groups of pictures and evaluated the DR grading of each fundus image. The sensitivity, specificity and diagnostic test consistency (Q-kappa value) of grading results were analyzed with the DR grading given by 3 senior ophthalmologists as the gold standard. The average Q-kappa values of 13 subjects were compared between the first four times and the last four times.Results:Through 8 round reading, the average Q-kappa was elevated from 0.67 to 0.81. Average Q-kappa of round 1 to 4 was 0.77, and average Q-kappa of round 5 to 8 was 0.81. The participants were divided into two groups. Participants in group 1 were junior ophthalmologists and participants in group 2 were medical students. Average Q-kappa of group 1 was elevated from 0.71 to 0.76. Average Q-kappa of group 2 was elevated from 0.63 to 0.84.Conclusions:The artificial intelligence reading label system was a useful tool in training junior ophthalmologists and medical students in doing diabetic retinopathy grading.

OBJECTIVE To investigate and analyze the bacteria on the nurse mask in ICU ward and its air effects disinfection.METHODS Sixty masks were examined for the bacteria numbers,germiculture and drug resistance.The bacteria numbers on the mask were recorded in 2,4,6 and 8 h with or without air disinfection,and the numbers of air bacteria were also estimated before and after air disinfection 2,4,6 and 8 h.The kinds and drug resistance of bacteria in the sputum of patients were recorded during the examining time.RESULTS The meticillin-resistant Staphylococcus(MRS,90.6%) and the Escherichia coli(ECO,9.4%) were the major bacteria on the mask.The numbers of bacteria on the mask at 2,4,6 and 8 h with air disinfection were 1.2?1.5,3.4 ?2.8,4.2?3.4 and 5.3?3.6 CFU/cm2,respectively.The numbers of bacteria on the air before air disinfection were 1264?314 CFU/m3 and after air disinfection 2,4,6 and 8 h were 917?213,790?176,676?179 and 483?167 CFU/m3,respectively.The numbers of bacteria on the mask and on the ward air were decreased obviously after the air disinfection comparing without disinfection.MRS was found in 26.7% patients sputum,and had the same drug-resistance spectrum as the bacteria on the mask.CONCLUSIONS MRS and ECO may be infected through air,and the effective air disinfection could reduce the numbers of bacteria on the air,and decrease the possibility of intercross infection.

Objective By detecting the DNA methylation and gene expression of long-chain 3-hydroxyacyl CoA dehydrogenase (LCHAD) in trophoblast cells, analyze the correlation of DNA methylation and gene expression in early-onset preeclampsia (EPE), hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome and antiphospholipid syndrome (APS), to investigate the molecular basis of long-chain fatty acid oxidation changes in different preeclampsia and pathological pregnancy. Methods Primary human cytotrophoblast cells and HTR8/Svneo cells were treated with serum from patients with EPE (14 cases), HELLP (12 cases), APS (14 cases), and normal pregnant women (NP, 14 cases). The methylation level of LCHAD gene promoter region through the MassARRAY platform and mRNA expression level by real-time fluorescent quantitative PCR technique were conducted. Results (1) Cytosine-phosphate-guanine (CpG)sites in human LCHAD DNA promoter region:CpG sites were detected in the range of 558 bp before LCHAD gene transcription start site, the detected CpG sites were 11 sites including 8 single sites and 3 complex sites. The position of these sites were at-984,-960,-899,-853,-811,-796,-774,-727,-615,-595,-579 respectively. (2) The sites of-899,-853,-615 and-595 showed increased methylation level in EPE and HELLP groups. The methylation level at-899,-853 and-615 sites in EPE and HELLP groups were significantly higher than those in NP group(P<0.01). The methylation level at-853 site was higher in EPE group than that in HELLP group(P<0.05). The-595 site showed the unmethylated in EPE, HELLP and APS groups. There were significantly difference between the 3 groups and EPE group(P<0.01). (3) The gene expression of LCHAD mRNA in EPE(0.048±0.005), HELLP(0.045±0.006)and APS(0.044±0.004) groups were significantly lower than NP group(0.076 ± 0.009;P<0.01). (4) The correlation of methylation level and gene expression in all groups: the methylation level at-899,-853,-727,-615 and-579 sites were negatively correlated with gene mRNA expression in EPE group (P<0.05). The methylation level at-899,-853 and-615 sites were negatively correlated with gene mRNA expression in HELLP group(P<0.05). Conclusions The variation of LCHAD DNA methylation of trophoblast cells are found among EPE, HELLP syndrome and APS. The different correlation of LCHAD DNA methylation and gene expression are different in pathological groups. LCHAD DNA methylation of EPE and HELLP syndrome were significantly increased and negatively correlated with LCHAD gene mRNA expression. These results further revealed the molecular basis of long-chain fatty acid oxidation in different preeclampsia and pathological pregnancy.

Objective To investigate the vasodilating effects of protocatechuic aldehyde (PCA ) on rat superior mesenteric arterial rings as well as its mechanism .Methods The tension of rat superior mesenteric arterial rings was recorded by a sensitive myograph system in vitro . We measured isometric tension changes in preconstricted rat superior mesenteric artery rings induced by potassium chloride (KCl ,60 mmol/L) ,serotomin (5-HT ,10-5 mol/L ) , and phenylephrine (PE , 10-5 mol/L ) after PCA treatment at different concentrations , respectively .We also observed vasodilating effects of PCA on KCl (60 mmol/L ) preconstricted rat superior mesenteric arterial rings after incubation with different inhibitors ,i .e .,L-NAME ,Indo ,ODQ ,4-AP (KV channel blocker) ,TEA (KCa channel blocker) ,BaCl2 (Kir channel blocker) ,and Glib (KATP channel blocker) ,respectively . Results PCA (10-6 -10-3 mol/L ) could relax KCl (60 mmol/L ) and 5-HT (10-5 mol/L ) preconstricted rat superior mesenteric arterial rings in a concentration-dependent manner . Indo of endothelial mechanism inhibitor blocked the vasodilating effect of PCA . 4-AP and BaCl2 of potassium ion channel inhibitors affected the vasodilatation induced by PCA in KCl (60 mmol/L)-preconstricted rat mesenteric artery .Conclusion PCA can relax KCl (60 mmol/L) ,or 5-HT preconstricted rat superior mesenteric arterial rings .This effect is associated with the inhibition of potassium channels and endothelial mechanism .

Objective:To evaluate the efficacy and safety of haploidentical(from family member donors) hematopoietic stem cell transplantation(HSCT) for children with hematologic malignancies.Methods: Fifty-eight children under fourteen years with hematological malignancies underwent haploidentical HSCT.All patients had poor-risk clinical or cytogenetic features and reclassified into high-risk and standard-risk groups.Transplantation outcomes were analyzed.Results: Of fifty-eight patient/donor pairs,seven(12.1%) were mismatched in two HLA loci,twenty(34.5%) in three loci,and thirty-one(53.4%) in four loci.Follow-ups were performed for a median of 1 663(752-2 664) days after transplantation.All patients achieved stable engraftments.The cumulative incidence of acute graft-versus-host disease(GVHD) of grades 2-4 was(54.8?7.6)%,and that of grades 3-4 was(11.4?4.8)%.The cumulative incidence of chronic GVHD was(45.6?7.8)% for the total and(19.6?6.5)% for the extensive.Thirty-eight patients survived with a 2-year actual overall rate of survival 59.0% and 78.9% in the high-risk and standard-risk group,respectively.The 3-year probability of LFS was(58.6?8.0)% and(68.4?10.7)%,respectively.Conclusion: The results encourage extending haploidentical HSCT without T-cell depletion treatments to children with an indication for transplantation.

The present study aims to investigate the effects of soluble endoglin (sEng) on invasive ability of cultured cytotrophoblasts of first trimester of pregnancy. Cytotrophoblasts of normal 6 to 8-week pregnancy were cultured by trypsin digestion method, and were incubated with cell culture medium without (control group) and with 10 μg/L sEng (sEng group), respectively for 24 h. The invasive ability was determined by transwell invasion assay, and expressions of MMP-2, MMP-9 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively. The results showed that the invasive ability of cytotrophoblasts in sEng group was lower than that in control group (P < 0.05). Compared with control group, the expressions of MMP-2 and MMP-9 mRNA and protein of cytotrophoblasts were significantly lower (P < 0.05). In conclusion, sEng may participate in the genesis of preeclampsia by affecting the invasive ability of cytotrophoblasts through regulation of the expression of MMP-2 and MMP-9.
Antigens, CD ; pharmacology ; Cell Movement ; drug effects ; physiology ; Cells, Cultured ; Endoglin ; Female ; Humans ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Placentation ; physiology ; Pre-Eclampsia ; physiopathology ; Pregnancy ; Pregnancy Trimester, First ; RNA, Messenger ; genetics ; metabolism ; Receptors, Cell Surface ; Trophoblasts ; cytology

BACKGROUNDPreeclampsia is a multifactorial disease during pregnancy. Dysregulated lipid metabolism may be related to some preeclampsia. We investigated the relationship between triglycerides (TGs) and liver injury in different preeclampsia-like mouse models and their potential common pathways.

METHODSPreeclampsia-like models (Nw-nitro-L-arginine-methyl ester [L-NAME], lipopolysaccharide [LPS], apolipoprotein C-III [Apo] transgnic mice + L-NAME, β2 glycoprotein I [βGPI]) were used in four experimental groups: L-NAME (LN), LPS, Apo-LN and βGPI, respectively, and controls received saline (LN-C, LPS-C, Apo-C, βGPI-C). The first three models were established in preimplantation (PI), early-, mid- and late-gestation (EG, MG and LG). βGPI and controls were injected before implantation. Mean arterial pressure (MAP), 24-hour urine protein, placental and fetal weight, serum TGs, total cholesterol (TC) and pathologic liver and trophocyte changes were assessed.

RESULTSMAP and proteinuria were significantly increased in the experimental groups. Placenta and fetal weight in PI, EP and MP subgroups were significantly lower than LP. Serum TGs significantly increased in most groups but controls. TC was not different between experimental and control groups. Spotty hepatic cell necrosis was observed in PI, EG, MG in LN, Apo-LN and βGPI, but no morphologic changes were observed in the LPS group. Similar trophoblastic mitochondrial damage was observed in every experimental group.

CONCLUSIONSEarlier preeclampsia onset causes a higher MAP and urine protein level, and more severe placental and fetal damage. Preeclampsia-like models generated by varied means lead to different changes in lipid metabolism and associated with liver injury. Trophoblastic mitochondrial damage may be the common terminal pathway in different preeclampsia-like models.

Animals ; Cholesterol ; blood ; Disease Models, Animal ; Female ; Fetal Weight ; physiology ; Liver ; injuries ; Male ; Mice ; Mice, Inbred C57BL ; Mitochondrial Diseases ; blood ; pathology ; Placenta ; metabolism ; Pre-Eclampsia ; blood ; pathology ; Pregnancy ; Triglycerides ; blood ; Trophoblasts ; pathology

Objective To study the efficacy and safety of human leukocyte antigen (HLA)mismatched hematopoietic stem cell transplantation (HSCT) on severe aplastic anemia(SAA). Methods From January 2006 to May 2010, 17 patients received mismatched HSCT. HLA antigens were 3-locimismatched in 9 patients, 2-loci-mismatched in 8. All patients received recombinant human granulocyte colony-stimulating factor (rhG-CSF) primed bone marrow cells plus peripheral blood stem cells after modified busulfan/cyclophosphamide + antithymocyte immunoglobulin (BU/CY + ATG ) conditioning regimen. Results All patients achieved full donor type engraftment. Grade Ⅲ-Ⅳ graft versus host disease (GVHD) occurred in 3 patients and extensive chronic GVHD in 1. With a median following-up time of 285(60-1670) d, 11 patients were alive, 9 of them had normal blood counts and the other 2 were blood transfusion independent. Six patients died of transplant-related complications. Conclusion Mismatched HSCT is a feasible and safe option for SAA patients without sibling identical donors.

Objective To analyse our series patients' data to assess its efficacy and safety of donor lymphocyte infusion (DLI) for Epstein-Barr virus (EBV) associated post-transplant lymphoproliferative disorders (PTLD) after allogeneic hematopoietic stem cell transplantation (HSCT). Methods Patients received HSCT from November 2006 to November 2009 and diagnosed as EBV associated PTLD by pathological or clinical methods were enrolled in this study. Lymphocyte was prepared by COBE collector.Related haplo-donors were the alternative if the original donors was unavailable. A range of mononuclear cell (MNC) dose of (0.5-1.0) × 108/kg was designed and the expected number of T lymphocyte included was at level of 107/kg. Cyclosporine (CsA) trough concentration was kept in a therapeutic level. Results Nine patients with PTLD received DLI 13 times, the median number of PBMC infused was 0.8 (0.16-1.03) ×108/kg, CD3+T cell number was 4.2 (1.6-5.7) × 107/kg. Seven patients received peripheral blood mononuclear cells (PBMC) from original haplo-identical donors, with 7 response and 6 complete remission.Defervescence occurred after 2 ( 1-5 ) d, and adenopathy began to recover in 6 ( 1-14 ) d after the initial infusion of leukocytes. Graft versus host diseases (GVHD) occurred in 6 recipients out of 7 evaluable patients, and all were controlled successfully. Three patients survived for 38, 23 and 3 months after PTLD.Conclusion In this small series cases, infusion of controlled dose of lymphocyte from primary donor is an effective and safe therapy for EBV associated PTLD after mismatched/haploidentical HSCT while the optimal regimen needs to be further studied.