Objective To investigat the changes of hemodynamics and arterial blood gas during the establishment of ECMO model and provide an ideal platform for the study of the mechanism of DCD protection using ECMO, a porcine model of controlled donors of cardiac death (CDCD) with extracorporeal support was established in the paper. Methods Eight domestic crossbred pigs were anes-thetized, atracurium (1mg/kg) and heparin (150~200 U/kg) was administered and the ventilator was discontinued while the animal was under deep anesthesia to kill the animal.Meanwhile, blood was drained from the left jugular vein catheter into the ECMO circuit and re-turned into the left femoral artery to establish the porcine model of CDCD with extracorporeal support . The average blood press and arterial blood gas were ministered at normal, cardiac arrest and ECMO supporting 1 h, 3 h and 5 h. Results The model of CDCD with extra-corporeal support was established successfully.Two animals restored the beating of the heart and the high doses of potassium chloride were given untill cardiac arrest during extracorporeal support.Blood gases throughout the apneic time showed a severe hypoxemia and acidosis, pH and PaO2 reduced to 7.09 ±0.24 and 5.3 ±0.93mmHg respective-ly, which can be improved during extracorporeal support.MAP and PaO2 were maintained over 300mmHg and 65mmHg respectively. Conclusion A new porcine model of controlled donors of cardiacdeath with extracorporeal support has been successfully established and showed to be feasible for related pathophysiological research work.

Objective To evaluate the effect of extracorporeal membranous oxygenation (ECMO) on the small bowel preservation in a porcine model of controlled donors after cardiac death (CDCD).Method The domestic crossbred donor pigs were anesthetized and ventilated with 100％ oxyger.An intravenous cannula was placed through iliac arteries and jugular vein,and connected to ECMO system.The ECMO was performed to infuse abdominal organs when cardiac death was declared.Pathology,electron microscope,energy metabolism and cell apoptosis level of intestinal mucosa were evaluated before operation,in cardiac death and at the end of extracorporeal support,respectively.Result The normothermic extracorporeal support could quickly restore graft blood supply and oxygenation.One-h extracorporeal support could improve the energy status in intestine of donation after cardiac death (DCD).The histologic damage and apoptosis of 1-h extracorporeal support had no significant difference with those before operation and in cardiac death (P＞0.05).With the extension of extracorporeal support,the intestinal mucosa damage degree was gradually increased,and the content of adenosine triphosphate in intestinal mucosa reduced gradually.Conclusion The normothermic extracorporeal support for 1 h in DCD is beneficial for improving the viability of small bowel.However,the integrity of intestinal mucosa is destroyed gradually as extracorporeal supporting time over,which may be initiated by the activation of intestinal epithelial apoptosis.

0.05).Conclusion The local relapse rate and the survival rate within 3 years has no significant difference between patients with lower rectal cancer of Duck stage A after sphincter-preserving surgery and Miles surgery.

Objective An ideal small bowel transplantation model is the prerequisite and guarantee of small bowel transplan-tation-related researches.The aim of this study is to establish a pig model of orthotopic segmental small bowel transplantation by modifi-cation of blood vessel reconstruction and surgical techniques. Methods Twenty healthy pigs were equally assigned as donors and re-cipients.The superior mesenteric vessels were freed and the distal ileum harvested as grafts.After irrigation and preservation, the small bowel was transplanted by end-to-end anastomosis of the superior mesenteric arteries and veins, and then the small bowel was re-constructed simultaneously to restore intestinal continuity, with the proximal part of the graft brought out as a stoma for the purpose of graft monitoring. Results Transplantation was successfully performed in 9 of the 10 cases, with a success rate of 90%.The donor operations averaged 100 min, recipient operations 172 min, warm ischemia 1.3 min, cold ischemia 96 min, and vascular anastomosis 27.6 min.No postoperative anastomosis fistula occurred in any of the cases. Conclusion The pig model of orthotopic segmental small bowel transplantation is ideal for small bowel transplantation-related researches.

Objective To develop a machine simultaneously integrating mechanized environmental cleaning and automatic mollusciciding and to evaluate its effectiveness of field application,so as to provide a novel Oncomelania hupensis snail control technique in the large?scale marshlands. Methods The machine simultaneously integrating mechanized environmental clean?ing and automatic mollusciciding,which was suitable for use in complex marshland areas,was developed according to the mech?anization and automation principles,and was used for O. hupensis snail control in the marshland. The effect of the machine on environmental cleaning and plough was evaluated,and the distribution of living snails was observed at various soil layers follow? ing plough. The snail control effects of plough alone and plough followed by mollusciciding were compared. Results The ma?chine could simultaneously complete the procedures of getting vegetation down and cut vegetation into pieces,plough and snail control by spraying niclosamide. After plough,the constituent ratios of living snails were 36.31% ,25.60% ,22.62% and 15.48% in the soil layers at depths of 0-5,6-10,11-15 cm and 16-20 cm respectively,and 61.91% living snails were found in the 0-10 cm soil layers. Seven and fifteen days after the experiment,the mortality rates of snails were 9.38% and 8.29% in the plough alone group,and 63.04% and 80.70% in the plough + mollusciciding group respectively(c27 d = 42.74,c215 d =155.56,both P values < 0.01). Thirty days after the experiment,the densities of snails were 3.02 snails/0.1 m2 and 0.53 snails/0.1 m2 in the soil surface of the plough alone group and the plough + mollusciciding group,which decreased by 64.92% and 93.60% ,respectively,and the decrease rate of snail density was approximately 30% higher in the plough + mollusciciding group than that in the plough alone group. Conclusions The machine simultaneously integrating mechanized environmental cleaning and automatic mollusciciding achieves the integration of mechanical environmental cleaning and automatic niclosamide spraying in the complex marshland areas,which provides a novel technique of field snail control in the large?scale setting in Chi?na.

Objective To study the relationship between phosphodiesterase 4D (PDE4D) gene rs966221 single nucleotide polymorphisms (SNPs) and ischemic stroke (IS) in Guangxi Zhuang population.Methods One hundred and one IS patients from Guangxi Zhuang Autonomous Region served as IS pgroup and 104 healthy subjects undergoing physical ecamination served as control group in this study.Their PDE4D gene rs966221 SNPs were detected by SNaPshot technique.The genotypes and frequencies of alleles were compared between the two groups and the relationship between PDE4D gene rs966221 SNPs and IS was analyzed.Results No significant difference was found in the GG,GA,AA genotypes and in the frequencies of G and A alleles between the two groups (0.99％ vs 3.85％,29.70％ vs 21.15％,69.31％ vs 75.00％,P＞0.05;15.84％ vs 14.42％,84.16％ vs 85.58％,P＞0.05).Univariate and multivariate logistic regression analysis showed that the PDE4D gene rs966221 SNPs were not related with the risk of IS in dominant AA vs GG+GA,recessive GG vs AA+GA and additive GG vs AA genetic models (P＞0.05).Conclusion The PDE4D gene rs966221 SNPs are not related with IS in Guangxi Zhuang population.

Chromosomal microdeletions and microduplications have been proven to be a significant proportion of genetic factors underlying birth defects. Chromosomal microarray analysis (CMA) and next generation sequencing-based copy number variation (CNV-seq) assay have been recommended as first-tier tests for prenatal evaluation of disease-causing CNV across the genome. With the broad application of such technologies in prenatal genetic diagnosis, there is a needed to enhance the consistency in interpretation and reporting of CNV results in clinical laboratories across China. In addition, a standard guideline for prenatal analysis and reporting of regions of homozygosity (ROH) is also required. To assist the classification, interpretation and reporting of CNV/ROH, the following recommendations have been developed, which may enhance a standard application of CMA/CNV-seq techniques in prenatal genetic diagnosis.

Genomic disorders caused by pathogenic copy number variation (pCNV) have proven to underlie a significant proportion of birth defects. With technological advance, improvement of bioinformatics analysis procedure, and accumulation of clinical data, non-invasive prenatal screening of pCNV (NIPS-pCNV) by high-throughput sequencing of maternal plasma cell-free DNA has been put to use in clinical settings. Specialized standards for clinical application of NIPS-pCNV are required. Based on the discussion, 10 pCNV-associated diseases with well-defined conditions and 5 common chromosomal aneuploidy syndromes are recommended as the target of screening in this consensus. Meanwhile, a standardized procedure for NIPS-pCNV is also provided, which may facilitate propagation of this technique in clinical settings.
Aneuploidy ; Cell-Free Nucleic Acids/genetics* ; Consensus ; DNA Copy Number Variations ; Female ; High-Throughput Nucleotide Sequencing ; Humans ; Pregnancy ; Prenatal Diagnosis

Objective:To study the expression of methyltransferase-like protein 14 (METTL14) in epithelial ovarian cancer and its clinical significance, and to explore the effect of METTL14 expression on the proliferation, invasion and migration of ovarian cancer cells.Methods:Immunohistochemistry (IHC) was used to detect METTL14 expression in tumor tissue samples, and analyze the relationships among METTL14 expression, clinicopathological factors, and prognosis in ovarian cancer. Lentiviral vectors and small interfering RNA (siRNA) were used to up-regulate and down-regulate the METTL14 expression in ovarian cancer cell lines A2780 and SKOV3, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was used to detect the N6-methyladenosine (m6A) content in ovarian cancer cells. Cell counting kit-8 (CCK-8), wound healing assay, and transwell assay were used to examine the function of METTL14 expression in the cells.Results:(1) The IHC score of METTL14 protein was 6.2±3.7 in 20 samples of ovarian cancer tissues and 3.3±2.5 in 15 samples of normal ovarian tissues, and the difference was statistically significant ( t=-2.64, P=0.012). Among the patients who suffered from ovarian cancer, there were 69 cases with high expression of METTL14 protein (IHC score≥6), accounting for 57.0% (69/121), and the cases with low expression of METTL14 protein (IHC score<6) accounting for 43.0% (52/121). Compared with the patients with low expression of METTL14, the patients with high expression of METTL14 had later stages, higher rates of lymph node metastasis, abdominal metastasis, and more ascite amount. The differences were statistically significant (all P<0.05). The overall survival rate was significantly lower in patients with high METTL14 expression than the low expression ( P=0.009). (2) LC-MS/MS data showed that the relative expression of m6A in A2780 and SKOV3 cells in the lentivirus (LV)-METTL14 group were 0.213±0.024 and 0.181±0.018, which were significantly higher than those in the LV-normal control (NC) group (0.109±0.022 and 0.128±0.020; all P<0.05). While the relative expression of m6A in A2780 and SKOV3 cells in the si-METTL14 group were 0.063±0.012 and 0.069±0.015, which were significantly lower than the expression in si-NC group of 0.108±0.014 and 0.121±0.014 (all P<0.05). CCK-8 assay showed that the absorbance values were significantly lower in the si-METTL14 group compared with the si-NC group at 36, 48, 60 hours (all P<0.05); while were significantly increased in the LV-METTL14 group compared with the LV-NC group at 48, 60 hours (all P<0.01). Scratch wound assays showed that the migration rate of the si-METTL14 group was lower than those of the si-NC group, while the LV-METTL14 group were higher than the LV-NC group by 24 hours, the differences were statistically significant (all P<0.01). Cell migration and invasion were detected by transwell migration and invasion assays. After cultivated for 24 hours, the invasion cell number and the migration cell number in the si-METTL14 group were less than those in the si-NC group. While the invasion cell number and the migration cell number in the LV-METTL14 group were more than those in the LV-NC group, respectively. The differences were statistically significant (all P<0.01). Conclusion:Patients with high METTL14 expression have a worse prognosis in ovarian cancer, which may increase the m6A modification of ovarian cancer cells and promote cells proliferation, invasion and migration.

With the extensive application of highly sensitive genetic techniques in the field of prenatal diagnosis, prenatal chromosomal mosaicisms including true fetal mosaicisms and confined placental mosaicisms are frequently identified in clinical settings, and the diagnostic criteria and principle of genetic counseling and clinical management for such cases may vary significantly among healthcare centers across the country. This not only has brought challenges to laboratory technician, genetic counselor and fetal medicine doctor, but can also cause confusion and anxiety of the pregnant woman and their family members. In this regard, we have formulated a consensus over the prenatal diagnosis and genetic counseling for chromosomal mosaicisms with the aim to promote more accurate and rational evaluation for fetal chromosomal mosaicisms in prenatal clinics.
Consensus ; Female ; Genetic Counseling ; Humans ; Mosaicism ; Placenta ; Pregnancy ; Prenatal Diagnosis/methods*