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 effect of preloading the epidural space with normal saline (NS) on the incidence of injury to blood vessel by epidural catheter placement for cesarean section. Methods One hundred and fifty parturients with a single baby at full term in vertex presentation scheduled for cesarean section under continuous epidural anesthesia were randomly divided into 3 groups ( n = 50 each): Ⅰ group control; Ⅱ group NS needle attached to a 5 ml syringe. Loss of resistance was used to identify the epidural space. In group Ⅰ no fluid was injected into the epidural space before insertion of catheter; while in group Ⅱ and Ⅲ NS 5 ml with or without whom blood or blood tinted fluid was withdrawn from epidural catheter was recorded. Results The number of patients in whom blood or blood tinted fluid was withdrawn from epidural catheter was significantly lower in group Ⅱand Ⅲ than in group Ⅰ but was not significantly different between group Ⅱ and Ⅲ. Conclusion Preloading the epidural space with 5 ml NS can reduce the incidence of injury to blood vessel induced by insertion of epidural

Objective To investigate the placental transfer and neonatal effects of dexmedetomidine during the cesarean section under general anesthesia.Methods Thirty-eight nulliparous parturients,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 22-37 yr,weighing 56-82 kg,who were at full term with a singleton fetus,scheduled for elective caesarean section under general anesthesia,were randomly divided into 2 groups (n=19 each) using a random number table:dexmedetomidine group (group D) and normal saline group (group N).In group D,dexmedetomidine was infused in a loading dose of 0.6 μg/kg starting from 10 min before induction of anesthesia,followed by an infusion of 0.4 μg · kg-1 · h-1 until peritoneal closure.Group N received the equal volume of normal saline.Blood samples were collected from the maternal artery (MA),umbilical vein (UV),and umbilical artery (UA) for blood gas analysis and for determination of plasma dexmedetomidine concentrations (CMA,CUV and CUA) by high-performance liquid chromatography-mass spectrometry,and CUV/CMA and CUA/CUV were calculated.Apgar scores were recorded at 1 and 5 min after delivery,and the occurrence of respiratory depression was also recorded.The I-D interval (the time from administration of anesthetics to delivery) and U-D interval (the time from incision of the uterus to delivery) were recorded.Results There were no significant differences in the blood gas analysis parameters in blood samples collected from the MA,UV and UA,I-D interval,U-D interval,and Apgar scores between the two groups (P＞0.05).No neonatal respiratory depression was found in both groups.In group D,CMA,CUV and CUA were 471±119,359±88 and (321±78) ng/ml,respectively,CUV/CMA was 0.76±0.06,and CUA/CUV was 0.89±0.03.Conclusion Although the metabolism of dexmedetomidine is little after easy placental transfer,dexmedetomidine has no adverse effects on the newborn during the cesarean section under general anesthesia.

OBJECTIVE:To provide reference for ration use of TCM injection. METHODS:The utilization of TCM injections in 39 hospitals from Chongqing area during 2012-2014 was analyzed statistically in respects of consumption sum,DDDs,DDC and B/A. RESULTS:The total consumption sum of TCM injections increased slowly in 39 hospitals from Chongqing area during 2012-2014,increasing from 767 581 thousand yuan to 911 759 thousand yuan,with annual growth rate of 8.98%. Total sum of consumption sum of top 10 drugs was 1 326 019.7 thousand yuan,accounting for 51.11% of total. The consumption sum of Hong-hua injection and Shenxiong glucose injection always took up the first 2 places in those years. DDDs of Xuesaitong injection, Shenxiong glucose injection glucose,Honghua injection and Shuxuetong injection always took up the first 4 places in those years. DDC of Xuebijing injection and Kanglaite injection always took up the first and second places in those years. Among top 10 TCM injection in the list of consumption sum,most of B/A was lower than 1.00;B/A of Shuxuetong injection,Xingnaojing injection, Shenqiong glucose injection and Xueshuantong injection were close or equal to 1.00. Xuesaitong injection was the one that B/A>1.00 from beginning to end. CONCLUSIONS:Total use of TCM injection increased steadily and drug use relatively concentrated in 39 hospitals from Chongqing area during 2012-2014,and rationality of drug use by physicians had been improved gradually.

Pregnancy ; Humans ; Female ; Cesarean Section/adverse effects* ; Abdominal Cavity ; Neoplasms

Purpose To investigate the feasibility to diagnose and evaluate the vulnerability of the atherosclerotic plaque through contrast-enhanced ultrasonography. Materials and Methods Thirty-four patients with carotid atherosclerosis were enrolled in the study to take contrast-enhanced ultrasonography and were observed on whether the plaque was enhanced and the features of its enhancement. The peak intensity (PI), time to peak (TTP), and density echo (DE) were calculated according to the time-intensity curve with QLAB software. Results The ratio of enhanced malacoplakia and the mixed plaque showed no difference (χ2=0.847, P>0.05). The percentages of enhanced plaque distributed on the base, tail and the shoulder were 70.0%, 23.3% and 6.7%, respectively with significant difference (χ2=29.100, P<0.001); the distribution ratio of enhanced plaque from high to low were as follows: the plaque on the shoulder > the plaque on the base> the plaque on the tail. It was positively correlated between enhanced plaque and its distribution (r=0.404, P<0.01). The TTP of the ROI between the malacoplakias and the mixed plaques showed no difference (t=0.479, P>0.05). The PI and DE of the ROI in the malacoplakia and the mixed plaques were analyzed by the time-intensity curve and the differences proved to be statistically significant (t=7.497 and 12.224, P<0.05). Conclusion Contrast-enhanced ultrasonography could present the neovessels in the atherosclerotic plaque, which is helpful in evaluating the vulnerability of atherosclerotic plaque.

Objective To explore the risk factors and optional treatment strategies for postoperative hypoxemia in patients undergoing acute Stanford A aortic dissection surgery.Methods From December 2012 to April 2014,108 patients received acute Stanford A aortic dissection surgery in Henan Provincial People' s Hospital.Of them,74 men vs.34 women,age (43.2 ± 10.1) years old,and weight (71.3 ± 18.4) kg.The operation was performed within 14 days since disease onset.All patients received surgery under deep hypothermic circulatory arrest(DHCA) and the nasal temperature goal was below 20 ℃ .Postoperative hypoxemia was defined as the arterial partial oxygen over inspired oxygen fraction(PaO2/FiO2) more than 200 after admission to ICU.Pulmonary protective ventilation management took place immediately after hypoxemia was diagnosed.Comparison analysis was performed between hypoxemia group and non-hypoxemia group in age,gender,body mass index (BMI),smoking history,duration of cardiopulmonary bypass(CPB),duration of DHCA,blood transfusion,length of mechanical ventilation,length of ICU stay and mortality.Results The incidence of postoperative hypoxemia was 40.7％ (44/108).There were no statistical differences in age,gender,hypertension,diabetes mellitus or COPD history.In the hypoxemia group,11 cases developed pulmonary infection,3 cases underwent tracheotomy and 10 cases died.In the non-hypoxemia group,4 patients died.Postoperative hypoxemia was significantly associated with more mortality,length of ventilation time,length of ICU stay and in hospital(P ＞ 0.05).Univariate analysis showed the associated factors were BMI,smoking history,preoperative hypoxemia,LVEF ＜ 0.45,operation time,aorta clamping time,DHCA time and blood transfusion.Multivariate analysis showed the independent predictive factors were preoperative hypoxemia,DHCA ＞40 min,blood transfusion ＞ 10 U and BMI ＞25 kg/ m2.Conclusion Obesity,massive blood transfusion,long DHCA time and preoperative hypoxemia are independent risk factors of postoperative hypoxemia in patients undergoing acute Stanford A aortic dissection surgery.Perioperative pulmonary protection should be paid more attention and reinforced.

ObjectiveTo evaluate the effect of preloading epidural space with 0.9％ sodium chloride on the incidence of the injury to blood vessel by epidural catheter insertion for cesarean section.Methods One hundred uterogestation patients with single birth,ASA class Ⅰ - Ⅱ,underwent caesarean section and requested continuous epidural analgesia were divided into group P and group C with each 50 cases by random digits table.After identification of the epidural space,group C was inserted epidural catheter directly,and 5 ml of 0.9％ sodium chloride was injected into epidural space through the epidural needle in group P,while the syringe plunger was held closed for 20 s to make sure the solution spreaded sufficiently,following insertion the epidural catheter.Between the two groups,mean arterial pressure and heart rate were recorded prior to anesthesia,2 min after turn to the supine horizontal position after succeeded puncture,the time when the fetus were born and when the surgery were over.The cases with bloody fluid in the epidural puncture needle during puncture,or in the epidural catheter during catheter placement,fresh blood in the epidural catheter,and bloody fluid in caudal end of epidural catheter during extubation were recorded.ResultsThe changes of mean arterial pressure and heart rate were all in the normal range,there was no obvious difference between the two groups.The incidence of rates with bloody fluid in the epidural puncture needle during puncture and the bloody fluid in caudal end of epidural catheter during extubation in group P were significantly lower than those in group C [ 10％ (5/50) vs.26％ (13/50),22％ (11/50) vs.48％ (24/50),P ＜ 0.05 or ＜ 0.01 ].ConclusionPreloading epidural space with 5 ml 0.9％ sodium chloride can reduce the incidence of the injury to blood vessel induced by insertion of epidural catheter.

OBJECTIVEThis paper aimed to determine the mRNA expression of osteoclast-related factors interleukin-6 (IL-6), interleukin-12 (IL-12) p35, IL-12p40, matrix metalloproteinase-9 (MMP-9), nuclear factor of activated T-cells cytoplasmic 1 (NFATcl), receptor activator of nuclear factor-KB (RANK), and tumor necrosis factor-α (TNF-α) mRNA in murine macrophages infected by a periodontitis patient's own tissue nucleic acid. Another aim was to investigate the effects of a periodontitis patient's own tissue nucleic acid on the differentiation of macrophages into osteoclasts.

METHODSInflammatory periodontal tissue samples of chronic periodontitis patients were taken during periodontal flap surgery, and healthy gingival tissue samples were taken from orthodontic patients during tooth extractions. Total RNA from periodontal tissue was extracted and reversely transcribed into cDNA and then cryo-preserved until further use. First, specific sequence oligodeoxynucleotide MT0I at a concentration of 1 µg · mL⁻¹ was added in murine macrophage RAW264.7, and the cells were incubated for 3 hours. Cells with PBS (1 µg · mL⁻¹) were used as negative controls. The inflammatory periodontal tissue cDNA and healthy periodontal tissue cDNA (1 µg · mL⁻¹) was added subsequently. There were four experimental groups: healthy periodontal tissue cDNA+ RAW264.7, inflammatory periodontal tissue cDNA+RAW264.7, MT01+healthy periodontal tissue cDNA+RAW264.7, and MT01+inflammatory periodontal tissue cDNA+RAW264.7. Real-time quantitative polymerase chain reaction was used to detect the mRNA expression of osteoclast-related factors IL-6, IL-12p35, IL-12p4O, MMP-9, NFATcl, RANK, and TNF-α mRNA after 3, 6, 12, and 24-hours.

RESULTSThe mRNA levels of osteoclast-related factors NFATc1, MMP-9, TNF-a, IL-6, IL-12p40, IL-12p35, and RANK in RAW264.7 were markedly upregulated with the treatment of periodontitis patient's own tissue nucleic acid. However, the mRNA expression of osteoclast-related factors was inhibited by use of an immunosuppressant MT01.

CONCLUSIONThe periodontitis patient's own tissue nucleic acid could promote the differentiation of murine macrophage into osteoclasts.

Animals ; Cell Differentiation ; Cytokines ; metabolism ; Gene Expression ; Gingiva ; Humans ; Interleukin-12 Subunit p40 ; Interleukin-6 ; Macrophages ; Matrix Metalloproteinase 9 ; Mice ; Osteoclasts ; metabolism ; Periodontitis ; RNA, Messenger ; Tumor Necrosis Factor-alpha

OBJECTIVETo investigate the therapeutic effect of IL-6 mAb on experimental autoimmune myocarditis (EAM) in rats, and search the mechanism of the role of IL-6, helper T cells 17 (Th17) and regulative T cells (Treg) in EAM pathogenesis.

METHODSThirty-four Lewis rats were divided into three groups randomly, i.e. control group (n = 6), EAM group (n = 12), and IL-6 mAb intervention group (n = 16). Rats in EAM group and IL-6 mAb intervention group were injected intracutaneously with myosin to establish EAM model. Rats in IL-6 mAb intervention group were injected intraperitoneally with 1 mg IL-6 mAb on 1st, 7th to 20th day after cardiac myosin immune injection. Myocardial inflammation was examined by HE stain, Masson stain, and TdT assay (TUNEL reaction) on 21st and 84th day after IL-6 mAb therapy in order to assess the therapeutic role. Spleen cells were analyzed by flow cytometry to illustrate Th17 and Treg cells? number and function. The serum concentration of IL-6, IL-10, IL-17, and TGF-beta in each group was measured by ELISA, concentration of STAT3, RORgammat, and Foxp3 mRNA in each group was determined with RT-PCR. Spleen cells derived from EAM were stimulated by IL-6 mAb in vitro, and the concentration of IL-10, IL-17 and TGF-beta was measured by ELISA.

RESULTSInflammation score, fibrosis score, and apoptosis index in IL-6 mAb intervention group were significantly decreased as compared with those in EAM group (P < 0.01). The number of Th17 and Treg cells in EAM group on the 21st day (experimental acute peak stage) were increased, and those in intervention group on the 21st day were significantly inhibited (P < 0.01). The concentration of serum IL-6, IL-10, IL-17 and TGF-beta in intervention group on the 21st day was decreased dramatically in comparison with that in EAM group on the same day (P < 0.01). The levels of peripheral blood STAT3, RORgammat, Foxp3 mRNA in intervention group on the 21st day was decreased significantly as compared with that in EAM group (P < 0.01). The expression of IL-10, IL-17 and TGF-beta was increased significantly (P < 0.01) by stimulation of IL-6 mAb on spleen cells derived from EAM in vitro.

CONCLUSIONSIL-6 mAb could neutralize IL-6, and ameliorate myocarditis and reduce heart autoimmune responses. IL-6 mAb has significantly protective effects on EAM by suppressing Th17 and Treg cells.

Animals ; Antibodies, Monoclonal ; therapeutic use ; Autoimmune Diseases ; drug therapy ; immunology ; Disease Models, Animal ; Forkhead Transcription Factors ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-17 ; metabolism ; Interleukin-6 ; immunology ; Male ; Myocarditis ; drug therapy ; immunology ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; metabolism ; Rats ; Rats, Inbred Lew ; STAT3 Transcription Factor ; metabolism ; Th17 Cells ; immunology ; Transforming Growth Factor beta1 ; metabolism