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 analyze the epidemic characteristics of multidrug-resistant tuberculosis(MDR-TB)in Hunan Province from 2013 to 2016,and provide theoretical basis for the prevention and control of tuberculosis.Methods Information about TB patients in Hunan Province reported by China Information System for Disease Control and Prevention between January 2013 and December 2016 was analyzed retrospectively.Results From 2013 to 2016,the total drug resistance registration rate in Hunan Province was 5.53/million(1 496/270 330 000),multidrug registration rate was 5.40/million(1 459/270 330 000),drug resistance rate and multidrug resistance rate showed an upward trends(trend x2 =113.605,96.590,respectively,both P<0.001).Among MDR-TB patients,male were more than females(74.09%vs 25.91%),most were more than 25 years of age,especially 45～age group(27.07%);the proportion of patients with MDR-TB retreatment was higher than that of the initial treatment(69.91%vs 30.09%).From 2013 to 2016,distribution range of MDR registration rates in different regions were 4.07/million-7.23/million.Conclusion MDR-TB in Hunan Province in 2013-2016 is increasing year by year,and mainly concentrate on young people over 20 years old.There are more cases of male and retreatment;it is necessary to strengthen regular treatment and prevention of key population,enhance the ability to identify and diagnose MDR-TB patients,and reduce the spread of MDR-TB.

Objective To assess the clinical efficacy and safety of the combination of intracoronary tirofiban infusion(ICTI) plus percutaneous coronary intervention(PCI) in patients with acute ST-elevation myocardial infarction (STEAMI). Methods The 128 cases with STEAMI were enrolled in this study. They were randomly divided into trial group and control group. The 10 μg/kg tirofiban were infused into the infarct related artery (IRA) within 5 minutes through the cather after coronary angiography in trial group (n=64). Normal saline in matched dose was infused into IRA after coronary angiography in control group (n=64). The coronary thrombosis and revascularization status were assessed within 10 minutes after injection. Postoperative bleeding complications were observed in all cases. Adverse cardiovascular events and cardiac function were followed up within 1 month in all cases. Results There were 33 cases whose thrombus burden was reduced within 10 minutes after the infusion of tirofiban in trial group, including 26 cases of thrombolysis in myocardial infarction (TIMI) ≥1. There were 6 cases whose thrombus burden was reduced within 10 minutes after the infusion of normal saline in control group, including 3 cases TIMI ≥ 1. The coronary thrombosis and revascularization status were better in trial group rather than in control group (P＜0.01). Adverse cardiovascular events occurred in 5 cases within 1 month, including 2 cases in trial group and 3 cases in control group (P＞0.05). New York heart association functional class and ejection fraction values were better in trial group rather than in control group within 1 month (P＜0.05). Postoperative bleeding complications occurred in 14 cases by TIMI criteria , including severe and mild bleeding in 2 cases in trial group and 1 cases in control group (P＞0.05), but no significant bleeding occurred in 8 cases in trial group and in 3 cases in control group (P＜0.01). Conclusions The combination of intracoronary infusion of tirofiban plus PCI is effective and safe for thrombolysis and revascularization in IRA in patients with STEAMI.

Objective To discuss the safety and operation effect of cesarean sectionof patients with early onset severe preeclampsia.Methods Clinical data were retrospectively analyzed,the treatment group made up of 236 patients with severe preeclampsia whose gestational age from 28 to 33 +6 weeks and the control group with 399 pregnancy women above 34 weeks.Main analytical items included:onset of gestational age,gestational age of termination of pregnancy,protensive of gestation,methods of termination of pregnancy,status of the occurrence of serious complications,fetal and neonatal mortality and the rate of neonatal asphyxia.Results The average conservative treatment time of treatment group was was significantly longer than that of control group [ ( 13.5 ± 1.2) d vs (9.6 ± 1.0 ) d,t =3.760,P ＜ 0.001].There was no significant difference on the cesarean section rate[70.8％ (167/236) vs 71.2％ (284/399),x2 =0.012,P =0.911 ] and neonatal mortality(2.5％vs 2.8 ％,x2 =0.026,P =0.871 ) between these two gourps.However there was significant difference on the rate of neonatal asphyxia(25.8％ vs 10.8％,x2 =20.792,P ＜ 0.001 ) and perinatal mortalit (22.0％ vs 6.9％,x2=27.782,P ＜ 0.001 ) between these two groups.Conclusion For the patients with severe early onset preeclampsia,to take conservative expectant treatment appropriately based on the safety of motherhood could avoid or reduce the possibility of the occurrence of serious complications,could extend the gestational age appropriately as well as reduce fetal and neonatal prevalence and mortality.Cesarean section is proper for the termination of pregnancy.

Objective To explore the application value of the modified hypotonic method of intestinal system contrast ultra-sonography in elder patients with small intestinal diseases .Methods Group A were 28 patients with small intestine disease and 8 patients who was found abdominal mass suspected source of intestinal tumors by routine abdominal ultrasound inspection .We con-trast the mannitol intestinal system contrast ultrasound with modified hypotonic method of intestinal system ultrasound contrast , and compare with gastrointestinal ,gastrointestinal barium meal contrast ,then contrast the results of pathology .Group B were 37 patients ,with definite diagnosis with duodenal ulcers ,polyps ,the descending part of the diverticulum ,who respectively treated with drinking water method and modified hypotonic method of intestinal system ultrasound contrast ,then carry the two methods into comparison .Results The nidus detection increase from 9 to 12 in group A ;the nidus detection increase from 13 to 33 in group B . The modified hypotonic method of intestinal system contrast ultrasonography ,which eliminate intestinal gas more obvious and relax the intestinal lumen more sufficient than conventional contrast ,can improve the nidus detection rate .Conclusion The modified hy-potonic method of intestinal system contrast ultrasonography could be as a routine inspection method for elder small intestinal dis-ease paitents .

Objective:To investigate the acceptance of advance care planning and its influencing factors in heart failure patients.Methods:A total of 208 patients with heart failure were surveyed by general data questionnaires and advance care planning acceptance questionnaires.Results:The total score of advance care planning acceptance of heart failure patients was (44.26 ± 11.73), the score of feeling dimension was (13.67 ± 5.72), the score of attitude dimension was (30.59 ± 6.33). 53.4%(111/208) of patients were willing to accept the talking about advance care planning. Regression analysis results showed that education level, New York Heart Association (NYHA) classification, communication status with medical staff and whether they had received life-sustaining treatment were important factors influencing of the acceptance of advance care planning in patients with heart failure.Conclusion:Patients with heart failure had higher acceptance of advance care planning. In clinical work, it is necessary to strengthen the scientific popularization of advance care planning in patients with low education level, low NYHA grade and no exposure to life-sustaining treatment. And strengthen the daily communication with patients to prepare for the follow-up advance care planning related communication.

Objective To study the clinical characteristics,muscle pathological features,diagnosis and prognosis of TK2-related mitochondrial DNA depletion syndrome(MDS).Methods Clinical and laboratory data of 2 cases of TK2-related myopathic MDS were reported.And data of previously reported 58 TK2-related MDS cases were reviewed.Results Total 60 patients consisted of 35 male and 25 female.The age of onset ranged from the birth to the age of 74 years old,and 54 of the patients were attacked at the age younger than 3 years old.Muscle weakness and hypotonia were detected in all patients,with 40 patients(including the newly diagnosed 2 cases) manifested as pure myopathic form,and 20 patients with other multiple organs involvement.Serum creatine kinase was mildly increased (211-6 500 IU/L) in 53 patients.Elevated serum lactic acid level (2.3-12.0 mmol/L)was observed in 24 patients.Muscle biopsy was available from 55 patients,and ragged red fibers and/or cytochrome C oxidase (COX)-negative fibers were detected in 48 out of them.Nine out of 11 patients received electronic microscope study showed proliferation of abnormal mitochondria.Respiratory chain enzymatic activities in skeletal muscle were reduced in 31 out of 33 patients.Marked mtDNA content reduction was observed in 36 out of 41 patients (4％-25％ of age-and tissue-matched controls).A total of 42 TK2 mutations were found in 60 patients,including 2 novel mutations c.923A ＞ G and c.619-2A ＞ T in this study.Conclusions The most common clinical manifestations of TK2-related MDS are severely,rapidly progressing myopathy with infantile or early childhood onset.As the detection rate of characteristic pathologic features in muscle is high,muscle biopsy is important for the diagnosis of TK2-related MDS.

ObjectiveTo investigate the correlation between the changes in routine blood test results and antiviral effect in patients with refractory chronic hepatitis C receiving standard treatment. MethodsA total of 63 previously untreated patients with refractory chronic hepatitis C who visited Beijing YouAn Hospital, Capital Medical University, from September 2011 to December 2012 were enrolled. The patients were treated with pegylated interferon-α-2a 180 μg/week and ribavirin (RBV) for 48 weeks. HCV RNA measurement and routine blood tests were performed at baseline and weeks 4, 12, and 24 of treatment to analyze the correlation between routine blood test results and antiviral effect. According to the status of sustained virologic response (SVR), the patients were divided into SVR group and nSVR group. The t-test was used for comparison of continuous data between groups, the chi-square test was used for comparison of categorical data between groups, and a simple linear correlation analysis was performed. ResultsOf the 63 patients, three were lost to follow-up, and the other 60 patients completed the antiviral therapy and 24-week follow-up; among the 60 patients, 46 achieved SVR, resulting in an SVR rate of 76.7%. Compared with the nSVR group, the SVR group showed lower absolute lymphocyte count (LYPH), white blood cell count (WBC), and absolute neutrophil count (NUET), and the three indicators were significantly different between the two groups at 12 weeks (t=3.398, 2.766, and 2.037, all P＜0.05); WBC and NUET still showed significant differences between the two groups at 24 weeks (t=2.559 and 2.151, both P＜0.05). In addition, at week 4 of treatment, the SVR group showed significantly greater reductions in these three indicators from baseline compared with the nSVR group, LYPH showed significant different between the two groups(t=226,P=003). At weeks 4 and 12 of treatment, LYPH, WBC, and NEUT were positively correlated with the reduction in HCV RNA (r=0.36, 045, 0.37, 0.47, 0.61, and 0.33, all P＜0.05). ConclusionThe reductions in LYPH, WBC, and NEUT can be used as predictive indicators for antiviral effect in patients with refractory chronic hepatitis C.

Objective:To develop a two-step method for purification of monoclonal antibody rhTF243 protein from mouse ascites by using hydrophobic charge induction chromatography(HCIC) and affinity chromatography with protein A sepharose CL-4B.Methods:The ascites was first purified by HCIC after centrifugation and filtration.Then the fraction containing the protein of interest was directly purified by affinity chromatography with protein A sepharose CL-4B.Results:The purity of the obtained monoclonal antibody was up to 97% with recovery of 73% and of high activity.Conclusion:The method for purification of monoclonal antibody is developed using HCIC and Protein A affinity chromatography and the obtained antibodies are of high purity and activity.

Objective To assess the value of peripheral blood thyroid stimulating hormone receptor(TSHR) mRNA determination in differential diagnosis of benign and malignant thyroid nodules.Methods Fine needle aspiration cytology (FNAC) and (or) postoperative histopathology as the gold standard were carried out,the expression of circulating TSHR mRNA was determined by RT-PCR in 33 patients with benign thyroid nodules,39 patients with thyroid cancer,and 20 normal controls.Results TSHR mRNA signals were not detected in normal controls,the positive rate of TSHR mRNA was higher in the group with malignant nodules than the group of benign nodules (91.2％ vs 48.5％,P＜0.01).TSHR mRNA level in the preoperative malignant group was significantly higher than that in the normal,benign,and postoperative cancer groups (all P ＜ 0.01).Using peripheral blood TSHR mRNA for differentiating benign or malignant of thyroid nodule had a sensitivity,specificity,and accuracy of 91.2％,51.5％ and 71.6％,respectively.The sensitivities of TSHR mRNA,FNAC,and these two methods combined in detecting malignant nodules were 91.3％,86.9％,and 100.0％ respectively,while diagnostic accuracies were respectively 84.0％,80.0％,and 92.0％.TSHR mRNA expression showed no significant relationship with sex,age,size,and number of nodule in these patients (all P ＞ 0.05),but it did exhibit significant difference between benign and malignant nodules(P＜0.01).Conclusion The peripheral blood TSHR mRNA could be used as a molecular marker for thyroid cancer,and it would help enhance the preoperative differentiation of benign and malignant thyroid nodules.