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

Chemically defined medium is widely used for culturing mouse embryonic stem cells (mESCs), in which N2B27 works as a substitution for serum, and GSK3β and MEK inhibitors (2i) help to promote ground-state pluripotency. However, recent studies suggested that MEKi might cause irreversible defects that compromise the developmental potential of mESCs. Here, we demonstrated the deficient bone morphogenetic protein (BMP) signal in the chemically defined condition is one of the main causes for the impaired pluripotency. Mechanistically, activating the BMP signal pathway by BMP4 could safeguard the chromosomal integrity and proliferation capacity of mESCs through regulating downstream targets Ube2s and Chmp4b. More importantly, BMP4 promotes a distinct in vivo developmental potential and a long-term pluripotency preservation. Besides, the pluripotent improvements driven by BMP4 are superior to those by attenuating MEK suppression. Taken together, our study shows appropriate activation of BMP signal is essential for regulating functional pluripotency and reveals that BMP4 should be applied in the serum-free culture system.
Animals ; Bone Morphogenetic Protein 4/metabolism* ; Cell Differentiation ; Chromosomal Instability ; Endosomal Sorting Complexes Required for Transport ; Mice ; Mitogen-Activated Protein Kinase Kinases/metabolism* ; Mouse Embryonic Stem Cells/cytology* ; Pluripotent Stem Cells/cytology* ; Signal Transduction ; Ubiquitin-Conjugating Enzymes

Objective To investigate the inhibition and possible mechanism of SAHA alone and combined with Er-lotinib on EGFR-TKI resistant lung cancer cell line H1975 cells. Methods The inhibitory effects of SAHA, Erlo-tinib and combined effect on cells were evaluated by CCK-8, colony formation assay and Transwell invasion assay. Western blot analysis was used to analyze the expression of PI3K, AKT, p-AKT, mTOR and p-mTOR protein in cells. Results SAHA and Erlotinib showed growth inhibition effects on H1975 cells with a dose-dependent man-nerand displayed synergistic inhibition effect. The combination of the drugs was significantly stronger than the single drug in the formation and invasion inhibition of tumor cells. The combined group had a strong inhibitory effect on the PI3K/AKT/mTOR signaling pathway. Conclusion SAHA combined with Erlotinib has synergistic inhibitory effects on the growth, cloning and invasion of EGFR-TKI lung cancer cell line H1975, which may be associated with inhibitory effects on the PI3K/AKT/mTOR signaling pathway.

Objective To analyze the risk factors and pathogen distribution of ventilator-associated pneumonia (VAP) in patients following coronary artery bypass grafting (CABG).Methods The clinical data of 1414 patients undergoing CABG in Teda International Cardiovascular Hospital from June 2014 to June 2016 were retrospectively analyzed .The VAP developed following CABG in 42 patients ( VAP group ) and not occurred in 1372 patients ( non-VAP group) .Multivariate logistic regression was used to analyze the risk factors of VAP in patients after CABG , and the microbial culture and drug sensitivity test were performed in VAP patients.Results The incidence rate of VAP after CABG was 2.97％ (42/1414).Multivariate logistic regression analysis showed that history of smoking ( OR =2.216, 95％ CI 1.018 -4.825, P <0.05), mechanical ventilation time >48 h (OR=7.457, 95％ CI 3.443-16.161, P<0.01), LVEF<40％(OR=3.524 , 95％ CI 1.203-10.325, P<0.05) and postoperative acute kidney injury (OR=16.239, 95％ CI 7.551 -34.924, P <0.01) were independent risk factors for VAP in patients after CABG.A total of 42 pathogen strains were detected in 42 patients with VAP, including 37 strains of Gram-negative bacteria, 2 strains of Gram-positive bacteria, and 3 strains of fungus.Gram-negative bacteria mainly were Klebsiella pneumoniae subspecies ( n =23, 54.76％) and Burkholderia cepacia ( n =6, 14.27％);the Gram-positive bacteria were Staphylococcus aureus ( n =2, 4.76％);the fungus was Candida albicans ( n =3, 7.14％).Klebsiella pneumoniae was sensitive to many antibiotics;and the resistance rate to amikacin , aztreonam , meropenem , and levofloxacin was <10％, the resistance rate to ceftazidime and piperacillin was <25％. Burkholderia cepacia was naturally resistant to amikacin , ampicillin, aztreonam, cefazolin, gentamicin and sulfamethoxazole .Conclusion The incidence of VAP was higher in patients after CABG , and the involved pathogens were mainly Gram-negative bacteria .Clinically , it is necessary to take necessary measures to prevent and treat VAP in order to improve the prognosis of patients undergoing CABG .

Objective To complete an information chain to support stomatological materials purchase and providing manage-ment.MethodsThe main problems of the materials management information chain were analyzed.The system construction,technical architecture and function design were designed based on B/S and C/S structure.Results The vendor cloud platform completed the information chain for purchase and distribution of stomatological materials,and provided technical support to hospital consumables management.Conclusion The system normalizes stomatological materials management,guarantees the safety,timeliness and accuracy of materials supply,and thus is worthy applying practically.

Objective To propose a method for equipment parameter demonstration to improve whole-life cycle management of equipment.Methods The measures and principles of equipment parameter demonstration were put forward based on its present situation and problems.The significance of equipment parameter demonstration for whole-life management was discussed from the aspects of biding,bidding evaluation and post-procurement management.Results The method enhanced equipment procurement,saved human resource,materials and economic cost,and solved the problems such as single source,unutilized equipment and high maintenance cost.Conclusion The method of equipment parameter demonstration maximizes the benefits,and promotes whole-life management of medical equipment.

Objective We investigated the value of endoscopic evacuation and craniotomy of the hypertensive in?tracerebral hemorrhage to determine which methods are more suitable for the patients. Methods One hundred twenty pa?tients with hypertensive intracerebral hemorrhage participated this study. They were divided into classic surgical evalua?tion group (n=60) and endoscopic surgical evaluation group (n=60) according to their corresponding surgery strategies. Each patient was assessed by the preoperative Glasgow Coma Scale (GCS), the mean rate and time of hematoma evacua?tion from onset to operation, the postoperative GCS, the mean time of admission in neuro-intensive care unit (NICU) and Glasgow Outcome Score (GOS) at 3 month after surgery. Results The continuous (≥3 months) follow-up surveys were all completed by 120 patients. There was no statistical difference in clinical data before operation between two groups (P>0.05). However, clearance of hematoma was much faster and more efficient in endoscopic surgical group than in classic surgical evaluation group (1.5 ± 0.4 vs.3.9 ± 0.6 h, P<0.01; 95.84 ± 2.72% vs.87.48 ± 7.84%, P<0.01). The GCS scores were 10(6,12),12(8,13) and 13(10,13) in endoscopic surgical group whereas were 6(5,9),7(5,11).8(5, 12) in craniotomy group at 1,3 and 7 d followed operation. GCS scores were higher in surgical group than in craniotomy group at all time points (P<0.01). In addition, patients receiving endoscopic treatment showed a shorter NICU admission time than those receiving craniotomy (3.55±4.21d vs. 9.10±4.72d, P<0.01). The intracranial infection and hypostatic pneumonia were sig?nificantly lower in endoscopic than in craniotomy surgery group (0 vs.6 cases; 5 vs. 41 cases, P<0.05). The endoscopic treatment significantly improved the GOS score compared with craniotomy [3(3, 4)vs. 2(2, 3)] (P<0.01). Conclusion Endoscopic evacuation of hematoma for hypertensive intracerebral hemorrhage is efficient and minimally invasive, which is superior to craniotomy.

Objective:To research on the genetic polymorphism distributions of fifteen short tandem repeat ( STR ) loci (D8S1179,D21S11,D7S820,CSF1PO,D3S1358,TH01,D13S317,D16S539,D2S1338,D19S433,VWA,TPOX,D18S51,D5S818, FGA) in Han race of Yunnan. Methods:A total of 313 specimens were collected from the unrelated individuals in Yunnan Han popu-lation. Genome DNA was extracted and amplified by multiplex PCR technique,the PCR products were analyzed by ABI-3130 genetic analyzer capillary electrophoresis detection, collected statistics of each STR loci genotypic frequency, and carried out the Hardy-Weinberg Genetic balance test. Results: No significant deviation from the Hardy-Weinberg Equilibrium was observed ( P>0. 05 ) , the heterozygosity of the fifteen STR loci in Yunnan Han population were found to be 0. 636-0. 901, Probability match was 0. 034-0. 220. Discrimination power of signal STR loci was 0. 780-0. 966, power of paternity exclusion was 0. 336-0. 797, polymorphism information content was 0. 555-0. 860,the combined accumulation discrimination power and exclusion probability for the 15 STR loci in Yunnan Han population were determined to be more than 0. 999 999 99 and 0. 999 998 408. The allele frequency of the 15 STR loci had a similarity compared with other areas in China,but also had a slight regional differences. Conclusion: The 15 STR loci( D8S1179, D21S11,D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, FGA ) demonstrate high genetic polymorphism in Yunnan Han population, they have a high forensic science application value in paternity testing and individual identification.

BACKGROUND:Neuromuscular junction structure has defects in patients with Duchenne muscular dystrophy, mainly presenting acetylcholine receptor structure fragmentation and the reduction of spine-like processes on thepostsynaptic membrane. It is generaly recognized that the structural defects are induced by structural damage of muscle cels and muscle fiber necrosis. OBJECTIVE:To explore the reasons of damage on neuromuscular junction in mouse models of Duchenne muscular dystrophy. METHODS: We introduced Duchenne muscular dystrophy models of male mdx mice and male Dko mice. After gene identification, they were used for tests. Male C57BL/6 mice were selected as normla controls. Hematoxylin-eosin staining was utilized to detect pathological changes in muscles. Neuromuscular junction structure was revealed using immunofluorescence staining. The differences in dystrophin expression, pathological features and neuromuscular junction structure were compared in mouse models of two kinds of Duchenne muscular dystrophy. RESULTS AND CONCLUSION:The introduced mouse models were accorded with the requirement of our experiment in aspects of genotype and protein expression levels. The number of acetylcholine receptor was apparently reduced in the neuromuscular junction of two kinds of mouse models. Although dko mouse muscles presented more obvious inflammatory infiltration and muscle fiber damage compared with mdx mice, but there was no significant difference in the damage to neuromuscular junction between them, and acetylcholine receptor fragmentation was identical. The evidence suggested that structural damage of neuromuscular junction and inflammatory pathological response are independent events. There is no direct relationship between them. Dystrophin gene deficiency is the main cause of the fragmentation of the acetylcholine receptor.

Objective To investigate the number, structure and demand of rehabilitation professionals hospitals above grade 2A in Jiangsu. Methods 176 hospitals above grade 2A in Jiangsu were investigated with questionnaire in the topics of general condition of department of rehabilitation medicine, structure of rehabilitation professionals and demand of rehabilitation professional. Results 64% of the hospitals has established department of rehabilitation. Rehabilitation professionals, such as the rehabilitation physician, therapists, nurses and engineer were in a total of 1657, and 231 professionals were needed further. Conclusion There are problems of lack of the department of rehabilitation medicine and rehabilitation professional, with the imbalance structure of professional in hospitals above grade 2A in Jiangsu.