Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.
Pregnancy ; Female ; Animals ; Mice ; Tetraploidy ; Blastocyst ; Embryo, Mammalian ; Cell Differentiation ; Embryonic Development

Objective:To observe the effect of selenium supplementation on 10-year survival rate of chronic Keshan disease (KD).Methods:The 10-year follow-up data of 302 patients with chronic KD at the KD surveillance sites in Shaanxi Province were collected from the Shaanxi Institute for Endemic Disease Control and Research and Xi'an Jiaotong University, 170 (56.3%) cases were given selenium supplementation (oral administration of sodium selenite tablet, once a week, 1 mg/time) until the end point of follow-up as selenium supplementation group, and the rest (132 cases) were non-selenium supplementation group. Cox proportional hazards models were used to identify the independent predictors for 10-year survival rate of chronic KD. Kaplan-Meier method was used to analyze the 10-year survival rate of patients with chronic KD during the follow-up period and the Log-rank test was used to compare the 10-year survival rate between groups.Results:The follow-up deadline was October 2019. During the follow-up period, a total of 199 patients (199/302, 65.9%) of chronic KD died, including 101 patients (101/170, 59.4%) in the selenium supplementation group and 98 patients (98/132, 74.2%) in the non-selenium supplementation group. In COX proportional hazards model, after adjustment for other baseline characteristics [age, sex, body mass index (BMI), family history of KD, smoking, blood pressure, heart rate, ECG abnormalities, initial cardiothoracic ratio, left ventricular ejection fraction (LVEF), and blood selenium content], selenium supplementation and combined use of angiotensin-converting enzyme inhibitor + β receptor blocker (ACEI + BBs) were protective factors for 10-year survival in patients with chronic KD (selenium supplementation: HR = 0.39, 95% CI: 0.28 - 0.53; ACEI + BBs: HR = 0.57, 95% CI: 0.39 - 0.84). The 10-year survival rate of chronic KD patients after selenium supplementation was significantly higher than that of non-selenium supplementation group (Log-rank test, P < 0.05). Conclusion:Selenium supplementation and combined use of ACEI + BBs in chronic KD patients, are associated with better survival during the 10-year follow-up.

We conducted the detection the Francisella spp.nucle acid from Hyalomma asiaticum asiaticum that main distribution is on railway line area from China-Kazakhstan border.The free-living ticks were collected and then identified by morphological and molecular methods.After species identification,they were detected by PCR targeting 16S rRNA and sdhA of Francisella spp.The amplified products were sequenced and the sequences was analyzed by using the Blast.A phylogenetic tree was constructed using MEGA 6 software.A total of 243 fleas were identified as H.asiaticum asiaticum.Only 35 samples were detected for Francisella spp.positive and the positive rate was 14.4％.Sequence analysis showed that two different sequences (seql and seq2) and all belong to Francisella-like endosymbionts (FLEs).Phylogenetic analyses showed that two FLEs were belong to the same cladd.This is first detection of FLEs nucleic acid from H.asiaticum Railway line area of China-Kazakhstan border.

Objective To investigate the distribution and drug resistance of pathogens in intensive care unit (ICU) so as to provide scientific basis for antibiotic adoption and the prevention and control of nosocomial infections. Methods The various specimens collected from the patients admitted into ICU in the First People's Hospital of Shunde Affiliated to the South Medical University from January 2007 to December 2014 were used to isolate the pathogens that might cause nosocomial infections and retrospectively analyze their clinical distribution and drug resistance. Kirby-Bauer paper diffusion and minimal inhibitory concentration (MIC) methods were applied to test the drug sensitivity, and according to National Committee for Clinical Laboratory Standards/Clinical and Laboratory Standards Institute (NCCLS/CLSI) standard, the results were identified.Results The sputum was the major specimen source in ICU, accounting for 68.8%, followed by urine (12.4%) and blood (6.8%). All together 557 pathogens in ICU causing nosocomial infections were isolated of which there were 377 gram-negative (G-) bacilli (67.7%), 103 gram-positive (G+) cocci (18.5%), and 77 fungi (13.8%). Among G- bacilli, the top three wereAcinetobacter baumannii (34.5%), Klebsiella pneumonia (17.8%), andPseudomonas aeruginosa (13.0%). Beside carbapenem, the drug resistance rates of Acinetobacterbaumannii to other antibiotics were more than 40%. The main G+ coccus causing nosocomial infection wasSaphylococcus aureus (36.9%) in ICU. The drug resistance rates ofSaphylococcus aureus to penicillin, gentamicin and erythromycin were higher than 50%. In 77 fungus strains,Candida albicans was ranked the first, accounting for 41.6%.Conclusion The main infection site in ICU is primarily respiratory tract, the G- bacilli are the predominate pathogens, and the drug resistance to antibiotics found in this report is serious, so clinically, the antibiotics should be properly used to avoid the occurrence of pathogenic strain with drug tolerance.

Objective To evaluate the effect of ultra short-term low-dose gamma knife radiation on invasive properties of glioma cells at the genetic level.Methods Malignant glioma cell lines U87 and U251 were treated with gamma knife radiation,respectively,with a marginal doses of 0,4,6,8 and 10 Gy (all to the 80％ isodose line).Real time-PCR was used to measure the mRNA expressions of AKT-1,AKT-2,β-catenin and TCF-4 at 30 min after radiation.Results The expressions of AKT-1,AKT-2,β-catenin,TCF-4 in glioma cell lines after low-dose gamma knife radiation with a marginal dose of 6-8 Gy were significantly increased as compared with those after 0 Gy (P＜0.05); reduced expressions ofAKT-1,AKT-2,β-catenin and TCF-4 after a marginal dose of 10 Gy were found as compared with those after 0 Gy (P＜0.05); the β-catenin and AKT-1 mRNA expressions in U87 cells and TCF-4,AKT-1 and AKT-2 mRNA expressions in U251 cells enjoyed the most obvious increase at a marginal dose of 6 Gy; the TCF-4 and AKT-2 mRNA expressions in U87 cells and β-catenin mRNA expression in U251 cells enjoyed the most obvious increase at a marginal dose of 8 Gy.Conclusion Low-dose gamma knife radiation (6-8 Gy) could increase the β-catenin,TCF-4,AKT-1 and AKT-2 expressions,while high dose (10 Gy) could inhibit these effects in a short time period of 30 min.

Carbonyl reductases catalyze carbonyl compounds to chiral alcohols that are important building blocks in fine chemical industry. To study carbonyl reductase from Pichia pastoris GS115 (ppcr), we discovered a new gene (ppcr) encoding an NADPH-dependent carbonyl reductase by genomic data mining. It was amplified by PCR from the genomic DNA, and expressed in Escherichia coli BL21 (DE3). The recombinant protein was purified to homogeneity. The optimum temperature was 37 degrees C and the optimum pH of PPCR was 6.0. PPCR was stable below 45 degrees C. The Km and k(cat) value of the enzyme for ethyl 3-methyl-2-oxobutanoate were 9.48 mmol/L and 0.12 s, respectively. The enzyme had broad substrate specificity and high enantioselectivity. It catalyzed the reduction of aldehydes, a-ketoesters, beta-ketoesters and aryl ketones to give the corresponding alcohols with >97% ee with only a few exceptions, showing its application potential in the synthesis of chiral alcohols.
Alcohol Oxidoreductases ; biosynthesis ; chemistry ; genetics ; Amino Acid Sequence ; Biotechnology ; methods ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Pichia ; enzymology ; Recombinant Proteins ; biosynthesis ; chemistry ; genetics ; Stereoisomerism ; Substrate Specificity ; Temperature

Asymmetric reduction of bulky diaryl ketones is still one of the challenging tasks in biocatalysis. By genomic data mining, a putative carbonyl reductase gene pascr was found in Pichia pastoris GS115. pascr was cloned and over-expressed in Escherichia coli Rosseta2 (DE3). The recombinant enzyme was purified to homogeneity by Ni-NTA column and its catalytic properties were studied. PasCR strictly used NADPH as cofactor, gel filtration and SDS-PAGE analysis suggested that the native form of PasCR was a dimmer. PasCR exhibited the highest activity at 35 degrees C in phosphate buffer at pH 6.5. The enzyme catalyzed the reduction of some bulky diaryl ketones, such as 4-methylbenzophenone, 2-methylbenzophenone and 4-chlorobenzophenone, especially for 4-methylbenzophenone, the product S--alcohol was obtained with 85% ee.
Alcohol Oxidoreductases ; genetics ; Amino Acid Sequence ; Catalysis ; Cloning, Molecular ; Ketones ; chemistry ; Molecular Sequence Data ; Pichia ; enzymology ; genetics ; Stereoisomerism

Objective To analyse the clinical effects on colon cancer with acute intestinal obstruction.Methods 56 patients with acute intestinal obstruction caused by colon cancer were studied retrospectively.The experience of diagnosis was summarized.Results 47 patients with acute intestinal obstruction caused by colon cancer received Stage Ⅰ tumor resection and the other 9 patients received different stages.54 patients healed (96.4％) and 1perioperative deaths (1.8 ％).Postoperative complications occurred in 18 cases (32.1％) including incision infection,intraperitoneal infection and intestinal fistula.Conclusion Stage Ⅰ tumor resection is feasible and safe surgical procedures for acute intestinal obstruction caused by colon cancer.

Objective To evaluate and improve the geometrical accuracy of the magnetic resonance imaging used for gamma knife radiosurgery localization. Methods Four common MRI scan programs were used in this study. The images were transferred to the Gamma Plan treatment planning system.Deviations between stereotactic coordinates based on MR images and real geometrical positions given by the construction of the phantom insert were evaluated for each study.The deviations were further investigated as a function of the spatial position of measured points in the investigated volume.Results The geometrical coordinate errors for axial and coronal images of the selected scan programs were all less than 1 mm.Optimal imaging procedure in terms of geometrical accuracy was observed using spin echo T1 and 3D FSPGR MRI sequences in the unit.Statistically significant differences were found in axial images amongSE T1,FSE T2,FRFSE Pd,FSPGR 3D sequences (F=1497.51,1873.63,2.87,3.00,P ＜0.05).The error increased along with the increase of the distance between node point and centricity.Conclusions The geometrical coordinate errors is related with the imaging program in M(il)l. Proper imaging programs and parameters should be selected according to the imageology character of patients' lesions when the radiosurgery is performed.

ObjectiveTo observe the effects of intravesical electric stimulation (IVES) on bladder sensation of neurogenic bladder.Methods28 patients with neurogenic bladder was evaluated, which all underwent intravesical bladder stimulation. Their bladder sensation were analyzed before and after the treatment.Results57.1% of the patients increased bladder sensation after treatment, 32.1% appeared the first sensation and urge sensation after treatment, 42.9% remained stable.ConclusionIntravesical bladder stimulation is effective to improve bladder sensation in a majority of the patients with neurogenic bladder.