Objective Coronary heart disease (CHD) is a multifactorial disease. This meta-analysis was performed to evaluate the relationship between angiotensinogen gene polymorphisms and CHD in the Chinese population. Methods We searched literature in pubmed (1990-2010.8) and CNKI (1990-2010.8) for all the relevant studies on 2 angiotensinogen polymorphisms (M235T and T174M) and risk of CHD. The meta-analysis software Stata 10.0 was used for ascertaining heterogeneity among individual studies and for combining all the studies. Furthermore,Egger's test and sensitivity analysis were performed to insure authenticity of the outcome.Results Ten associations studies on 2 angiotensinogen polymorphisms (M235T and T174M) were included in this meta-analysis. In a combined analysis, the summary per-allele odds ratio for CHD of the M235T polymorphism was 1.374 (95% confidence interval, 1.019 to 1.852) and T174M polymorphism was 4.089 (95% confidence interval, 1.697 to 9.851). Conclusions The M235T polymorphism had weak but statistically significant association with CHD while the T174M polymorphism was more strongly associated with a CHD risk in Chinese population, but further confirmation studies are needed.

Interleukin 33 (IL 33),one member of the IL-1 family,is expressed in many types' tissue and regulation of multiple target cells via its suppression of tumorigenicity 2 (ST2) receptor.Therefore,the crucial roles of the novel cytokine IL-33 in allergic,endocrine diseases,infectious diseases and cancer are becoming characterized.The function of IL-33 in different parasite infection is distinctive in parasitic infections,due to the difference in pathogenic mechanism and in the time course of IL-33 expression.

Objective To investigate the expressions of c-Fos,5-HT and spontaneous activities in single prolonged stress (SPS) stress rats,and to study the possible mechanism of posttraumatic stress disorder(PTSD).Methods forty-eight male SD rats were randomly divided into 6 groups:Individual living + SPS group (IS group),Individual living+control group (IC group),Enriched environment+SPS group (ES group),Enriched environment+control group (EC group),control+SPS group(CS group),control group(C group),8 rats in each group,spontaneous activities were measured before the experiment and post experiment at week 1,2.And the expressions of c-Fos,5-HT in the hippocampus were examined by immunohistochemical staining.Results (1) Spontaneous activity:there were no differences of crossing number and distance among 6 groups before experiment (P＞0.05).On SPS 14 days the crossing number and distance of IS group (12.12±9.64,(2.71 ± 1.99)m) decreased compared with the CS group(45.25±8.37,(6.37± 1.18) m,P＜0.05),and ES group (69.75± 10.05,(10.69± 1.50) m)showed significant increase compared with CS group (P＜0.05).(2)5-HT:the expression of 5-HT in the hippocampus of IS group(0.1125±0.0095) was significantly higher than that in CS group(0.6138±0.0059,P＜0.05),menwhile lower in ES group(0.0495±0.0074,P＜0.05).(3)c-Fos:compared with CS group(0.3108±0.0074),the expression of c-Fos in the hippocampus of IS group(0.3585±0.0150,P＜0.05) significantly increased,but decreased in ES group (0.2613±0.0063,P＜ 0.05).Conclusion Enriched environmental can improve the level of activities in PTSD rats,and to reduce the the expression of c-Fos and 5-HT in the hippocampal neurons.

Objective:Explored the clinical application value of the current reference interval of serum creatinine (using the 2015 health industry standard) and the original reference interval (using the third edition of the clinical laboratory operating procedures).Methods:This paper is a retrospective study. The study collected 360 normal serum samples of the adults who visited the First Hospital of Hebei Medical University from February to April 2019 and,aged 20-79 years old (including 90 males aged 20-59, 90 females aged 20-59, 90 males aged 60-79, and 90 females aged 60-79). The concentration of cystatin C was determined in the above specimens, combined with the concentration of creatinine, and the results calculated by the Glomerular Filtration Rate Estimation Formula (eGFR) was used as the standard for judgment. In this study, the chi-square test was used to statistically analyze the agreement between the results of the new and old reference intervals of creatinine and the results calculated by the eGFR formula.Results:The total coincidence rate of Scr and eGFR in 360 adults aged 20-79 years was 83.40% in the current RI, and 77.00% in the original RI. The coincidence rate of the former was significantly higher than that of the latter ( P=0.03). The positive coincidence rate of Scr and eGFR (214 patients with eGFR positive) was 91.86% in current RI, 41.59% in the original RI, the coincidence rate of the former was significantly higher than that of the latter ( P<0.01). The negative coincidence rate of Scr and eGFR (146 patients with eGFR negative) was 74.66% in current RI, and 99.32% in original RI, the coincidence rate of the latter was significantly higher than that of the former ( P<0.01). Consistent results were shown in groups of males aged 20-59, females aged 20-59 and females aged 60-79. However, in the group of males aged 60-79 years, the total coincidence rate of Scr and eGFR was 75.56% in current RI, and 88.89% in original RI, the coincidence rate of the latter was significantly higher than that of the former ( P =0.01); positive coincidence rate of Scr and eGFR was 57.69% in current RI, and 98.08% in original RI, the coincidence rate of the latter was significantly higher than that of the former ( P<0.01); negative coincidence rate of Scr and eGFR was 100.0% in current RI, and 76.32% in original RI, the coincidence rate of the former was significantly higher than that of the latter ( P<0.01). Conclusions:The current reference interval is better than the original reference interval in screening for decreased glomerular filtration rate, and it is favorable for the early detection of patients with mildly decreased renal function. However, among males aged 60-79 years old, the accuracy of the current RI and the ability to screen for decreased glomerular filtration rate are inferior to the original RI, which needs to be further observed and studied.

Objective During pregnancy , exosomes can be released from the placenta into maternal circulation and play im-portant roles in normal pregnancy or placenta-related diseases .We aimed to establish a simple and efficient method for isolating and i-dentifying placental exosomes from maternal serum and lay a foundation for the studies of pregnancy -related diseases . Methods Using sucrose gradient centrifugation with 8% PEG6000 precipitation twice , we isolated and purified placenta-derived exosomes from normal maternal serum and detected their molecular markers CD 63 , CD81 and PLAP by Western blot , followed by silver staining anal-ysis of the protein profile of the exosome pellet .We identified the morphology of the placenta-derived exosomes by transmission electron microscopy ( TEM) and measured the size and distribution of the particles by dynamic light scattering ( DLS) . Results Silver stai-ning of the protein profiles of the exosomes after sucrose gradient centrifugation clearly revealed the bands of the protein molecules . Western blot showed the expressions of CD 63, CD81, and PLAP in the 21-34%density layer, which demonstrated the presence of serum placental exosomes mainly in the 1.09-1.16 g/mL density layer.TEM exhibited that the placenta-derived exosomes were round or oval cup-shaped, specifically expressing PLAP, and the particles were uniform in size, with a mean diameter of (41.79 ±11.94) nm. Conclusion A simple, fast, and efficient method was successfully established for isolating placenta-derived exosomes from ma-ternal serum, which provides a basis for studying the roles of placental exosomes in normal pregnancy and placenta -related diseases.

This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G(1) phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G(1) phase. L929 cells were sensitive to s-TNF-α when synchronized in G(1) phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G(1)/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.

Objective To study the risk factors of lymph node metastasis in the central neck compartment of thyroid carcinoma,and to explore the reasonable range of lymph node dissection in central neck dissection for clinically node-negative papillary thyroid microcarcinoma patients.Methods From Dec.2015 to Dec.2016,a total of 200 patients with CN0 papillary thyroid carcinoma were randomly divided into two groups according to the registration number:unilateral central neck dissection group and bilateral central neck dissection group in Department of Thyroid Surgery,Fujian Medical University Union Hospital.The risk factors of lymph node metastasis and value of bilateral central neck dissection were analyzed.Results The risk factors of lymph node metastasis in the central papillary thyroid carcinoma were ≥0.7 cm in diameter and older than 45 years in age and gender in male.Further analysis found that contralateral central lymph node metastasis occurred in patients with tumor diameter ≥0.5 cm.The positive rate was 22％.The number of lymph nodes detected in the unilateral and bilateral central areas was 9.53±6.04 and 12.19±7.18,P=0.035,respectively.The positive numbers of lymph nodes were 1.17±1.47 and 2.11±2.75,P=0,022 respectively.Conclusion In patients with tumor diameter ≥0.5 cm,bilateral central neck dissection is conducive to improving the thoroughness of tumor dissection and does not increase the risk of complications.

Objective To explore the predictive value of serum CEA and cytokeratin-19 fragments (CYFRA21-1)prior treatment for the epidermal growth factor receptor (EGFR) mutation and efficacy of tyrosine kinase inhibitors ( TKI ) in patients with non-small cell lung cancer.Methods The study was a clinical research.Totally 101 matched tissue and plasma samples were collected from Peking University People′s Hospital from 2012 to 2013.All clinical specimens were analyzed for EGFR mutations in exons of 18, 19, 20 and 21 by ADx-ARMS and direct sequencing, and the serum levels of CEA and CYFRA21-1 were analyzed by ECLI.The correlation between EGFR mutant status and efficacy of EGFR-TKI and clinicopathological parameters were analyzed by χ2 test, Log-rank text and Cox proportional hazards regression model.Results The mutation rate was 60.4%(61/101) by ADx-ARMS and 33.7%(34/101) by direct sequencing.Mutations were more frequently observed in the higher serum CEA level patients(≥5μg/L,78.8%).However, the rates of EGFR mutations of different CEA levels were similar.Among the patients receiving TKI therapy, the efficacy of EGFR-TKI was closely related to serum CYFRA21-1 level prior treatment and EGFR mutation (χ2 =8.903, P =0.003; χ2 =28.590, P <0.001 ).And serum CYFRA21-1 level prior treatment and EGFR mutation were independent factors for EGFR-TKI treatment affecting PFS (RR=0.298, P<0.001;RR=0.086, P<0.001).Conclusion The mutation rate of EGFR was significantly related with the expression level of CEA prior treatment, and serum CEA and CYFRA21-1 levels prior treatment could be potential predictors of EGFR-TKI efficacy.

OBJECTIVETo establish a canine cell line with p53 gene knockdown by lentivirus- mediated RNA interference (RNAi).

METHODSFour pairs of oligonucleotide sequences of p53 gene were synthesized and cloned into the pGMLV-SC5 RNAi vector. Following confirmation by PCR and DNA sequencing, the recombinant pGMLV-p53 plasmids and packaging mix vectors were packaged into mature lentivirus to infect 293T cells. The supernatant of the infected cells was harvested to infect WRD cells, in which p53 expression was detected using Western blotting and real-time PCR. The lentivirus with the strongest p53⁻ silencing effect was packaged for infecting WRD cells at the optimal multiplicity of infection (MOI) to establish a stably infected cell line (WRD/p53⁻) screened using puromycin.

RESULTSThe lentivirus carrying p53 shRNA was constructed successfully and a virus titer of 1 × 10⁹ TU/ml. The 4 pGMLV-p53 plasmids all produced strong p53 interference affects, and pGMLV- p53A1 with the strongest effect. The stably infected cells line WRD/p53⁻ was established successfully using pGMLV-p53A1 plasmid.

CONCLUSIONThe canine cell line WRD/p53⁻ with stable lentivirus-mediated p53 silencing has been established successfully.

Animals ; Cell Line ; Dogs ; Gene Knockdown Techniques ; Genes, p53 ; Genetic Vectors ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction

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