Understanding the regulatory networks for germ cell fate specification is necessary to developing strategies for improving the efficiency of germ cell production in vitro. In this study, we developed a coupled screening strategy that took advantage of an arrayed bi-molecular fluorescence complementation (BiFC) platform for protein-protein interaction screens and epiblast-like cell (EpiLC)-induction assays using reporter mouse embryonic stem cells (mESCs). Investigation of candidate interaction partners of core human pluripotent factors OCT4, NANOG, KLF4 and SOX2 in EpiLC differentiation assays identified novel primordial germ cell (PGC)-inducing factors including BEN-domain (BEND/Bend) family members. Through RNA-seq, ChIP-seq, and ATAC-seq analyses, we showed that Bend5 worked together with Bend4 and helped mark chromatin boundaries to promote EpiLC induction in vitro. Our findings suggest that BEND/Bend proteins represent a new family of transcriptional modulators and chromatin boundary factors that participate in gene expression regulation during early germline development.
Animals ; Cell Differentiation/genetics* ; Chromatin/metabolism* ; Embryonic Stem Cells ; Germ Cells/metabolism* ; Germ Layers/metabolism* ; Mice

Objective: To explore the relevant gene, signaling pathway for permanent atrial fibrillation (pAF) occurrence in order to provide the molecular basis of the pathogenesis of pAF. Methods: Our research included in 2 groups: pAF group, n=7 patients and Control group, n=4 healthy subjects with sinus rhythm. Agilent 4x44K microarray was used to analyze the mRNA in left atrium for differential gene expression profile. Based on Gene Ontology, KEGG and Biocarta databases, differentially expressed genes were studied for their relevant function and signaling pathway. Furthermore, the genes with significant differences were verified by quantitative real time PCR (qRT-PCR) in pathological specimen from 5 pAF patients and 5 normal heart donors. Results: The expression profile identified 987 abnormally expressed genes, 567 of them were down-regulated and 420 were up-regulated. 9 genes with significant differences were verified by qRT-PCR in pathological specimen and the changes were similar to microarray; those genes were closely related to pAF by involving left atrium fibrosis, electrical remodeling, inflammation, cellular stress response, metabolism and transcription regulation. GO and Pathway analysis indicated that down-regulated genes were mainly involved in metabolic processes; up-regulated genes had the effects on cellular stress response, immune response and platelet activation. Conclusion: Microarray technology identified some important genes related to pAF occurrence; such genes involved in left atrial structural and functional remodeling via affecting cellular metabolism, inflammation, immune response and thrombogenesis in relevant patients.

Quantitative models were established to analyze the content of chlorogenic acid and soluble solid content in the liquid-liquid extraction of Reduning injection by near-infrared (NIR) spectroscopy. Seven batches of extraction solution from the liquid-liquid extraction of Lonicerae Japonicae Flos and Artemisiae Annuae Herba were collected and NIR off-line spectra were acquired. The content of chlorogenic acid and soluble solid content were determined by the reference methods. The partial least square (PLS) and artificial neural networks (ANN) were used to build models to predict the content of chlorogenic acid and soluble solid content in the unknown samples. For PLS models, the R2 of calibration set were 0.9872, 0.9812, RMSEC were 0.1533, 0.7943, the R2 of prediction set were 0.9837, 0.9733, RMSEP were 0.2464, 1.2594, RSEP were 3.25%, 3.31%, for chlorogenic acid and soluble solid content, respectively. For ANN models, the R2 of calibration set were 0.9903, 0.9882, RMSEC were 0.0974, 0.4543, the R2 of prediction set were 0.9868, 0.9699, RMSEP were 0.1920, 0.9427, RSEP were 2.61%, 2.75%, for chlorogenic acid and soluble solid content, respectively. Both the RSEP values of chlorogenic acid and soluble solid content were lower than 6%, which can satisfy the quality control standard in the traditional Chinese medicine production process. The RSEP values of ANN models were lower than PLS models, which indicated the ANN models had better predictive performance for chlorogenic acid and soluble solid content. The established method can rapidly measure the content of chlorogenic acid and soluble solid content. The method is simple, accurate anc reliable, thus can be used for quality control of the liquid-liquid extraction of Reduning injection.
Chlorogenic Acid ; analysis ; Drugs, Chinese Herbal ; analysis ; Injections ; Least-Squares Analysis ; Liquid-Liquid Extraction ; standards ; Neural Networks (Computer) ; Quality Control ; Spectroscopy, Near-Infrared ; methods

Alveolar epithelial type II (AT II) cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells (fibroblasts). Progenitor cells are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A (SPA) suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase (rAAV-SPA-TK) system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells (H441) transfected with TK gene had higher sensitivity to ganciclovir (GCV) than SPA low expression cells (A549). In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell (AT II) niche in vitro and in vivo.
Antiviral Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cells, Cultured ; Dependovirus ; genetics ; Dose-Response Relationship, Drug ; Electrophoresis, Polyacrylamide Gel ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Ganciclovir ; pharmacology ; Gene Expression Regulation, Neoplastic ; Genes, Transgenic, Suicide ; genetics ; Genetic Vectors ; genetics ; Humans ; In Situ Nick-End Labeling ; Luciferases ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics ; Pulmonary Alveoli ; cytology ; metabolism ; Pulmonary Surfactant-Associated Protein A ; genetics ; metabolism ; Thymidine Kinase ; genetics ; metabolism

Alveolar epithelial type II (AT II) cells are essential for lung development and remodeling, as they are precursors for type I cells and also produce other non-repair cells (fibroblasts). Progenitor cells are believed to possess capability of multi-potent transdifferentiation, which is closely related to the niche, suggesting the importance of establishment of a lung progenitor cell niche model. We hypothesized that pulmonary surfactant-associated protein A (SPA) suicide gene system would cause AT II cell to kill itself through apoptosis and leave its niche. In vitro, the recombinant adeno-associated virus vectors-SPA-thymidine kinase (rAAV-SPA-TK) system was established to get targeted apoptotic AT II cells. The apoptosis of AT II cells was detected by using MTT. The results showed that cloned SPA gene promoter had specific transcriptional activity in SPA high expression cells, and SPA high expression cells (H441) transfected with TK gene had higher sensitivity to ganciclovir (GCV) than SPA low expression cells (A549). In vivo, increased apoptosis of AT II cells induced by GCV in rAAV-SPA-TK system was observed by TUNEL. Finally, the successful packaging and application of rAAV-SPA-TK system provide experimental basis to get specific lung progenitor cell (AT II) niche in vitro and in vivo.

Background Selective laser trabeculoplasty(SLT)has been widely accepted for the treatment of open-angle glaucoma.However,there are few reports about its application in treating angle-closure glaucoma so far.Objective This study was to clinically evaluate the safety and efficacy of selective laser trabeculoplasty for primary angle-closure glaucoma with opened anterior angle follow-up surgery.Methods A case-observational design was adopted in this study.A total of 33 eyes from 23 angle-closure glaucomatous patients with opened 180° angle were enrolled.All the patients understood this trial and written informed consent was obtained prior to the medical treatment.The average preoperative intraocular pressure(IOP)and number of medication were(23.07±4.01)mmHg and 1.52±0.66,respectively.A frequency doubled Q-switched Nd:YAG laser was used to irradiate the trabecular meshwork with 70-80 laser spots over 180°.The same preoperative anti-glaucoma drugs were topically administered during the follow-up period.Visual acuity,IOP,slit lamp examination,gonioscopy and the coefficient of outflow facility were regularly assessed and performed after laser surgery.Results The IOP values of the 33 eyes were(18.78±4.87)mmHg,(18.83±3.95)mmHg,(19.78±4.48)mmHg,(19.63±3.68)mmHg,(19.11±3.36)mmHg,and(21.17±3.14)mmHg 1 day,l week,and 1,3,6,9 months after operation,showing significant decline in comparison with preoperation(P＜0.01).The mean drug uses were 1.63±0.85,1.23±1.01,1.25±0.98,1.21±1.02,1.22±0.86,and 1.69±0.53 at the same time point after operation,respectively,and significant differences were found 1 week,1,3,6 months in postoperation in comparison with preoperation(P =0.020,0.026,0.020,0.031).Ciliary congestion and aqueous flare naturally disappeared 1 week after surgery.The coefficients of outflow facility were 0.11±0.09 and 0.23±0.15,respectively at pretreatment and 3 months after SLT with a statistically significant difference (P=0.002).Conclusions Selective laser trabeculoplasty is effective and safe for angle-closure glaucoma.However,it is essential to have 1/2 circle of anterior chamber open for SLT in angle-closure glaucoma.

Objective To study the effect of new appointment service model on improving the efficiency of diagnosis and treatment of large comprehensive hospitals.Methods All patients seeing doctors at out-patient department were designated as experimental group that implemented new appointment service model of diagnosis and treatment during July to August of 2011,while the patients at out-patient department were designated as control group that implemented routine service model during May to June of 2011.Then,differences in appointment time,non-effective appointment rate and patients' satisfactory rate between these two groups were compared.Results In experimental group,X-ray appointment examination time:68.8％ patienta were ≤ 1 day and 31.2％ patients were ≥2 days,ultrasound appointment examination time:67.2％ patients were≤1 day and 32.8％ patients were ≥ 2 days,endoscope inspection appointment examination time:85.6％ patients were ≤2 days and 14.4％ patients were ≥ 3 days;appointment examination time of cardiovascular ultrasound:75.9％patients were ≤ 2 days and 24.1％ patients were ≥ 3 days ; and those significantly different with that of control group ( x2 =4.838,9.199,28.355,10.359,respectively; P ＜ 0.05).Non-effective appointment rate of experimental group was that radiology department 1.9％,ultrasound department 2.4％,endoscope inspection room 4.1％,cardiovascular ultrasound 2.5％,and significantly lower than that of the control group ( x2 =9.982,19.687,8.684,7.623,respectively; P ＜ 0.05 ).The patients' satisfactory rate of experimental group was that:radiology department 94.9％,ultrasound department 93.6％,endoscope inspection room 96.3％,cardiovascular ultrasound 95.80％.And the parameters of experimental group were obviously better than that of control group ( x2 =30.623,4.120,15.640,19.231,respectively; P ＜ 0.05 ).Conclusions Through implementing new appointment service model of diagnosis and treatment,the patients' appointment time was shortened,non-effective or repeatedly-changed appointment was decreased,and the efficiency of diagnosis and treatment and the patients' satisfactory rate were improved obviously.

OBJECTIVETo investigate the changes of thyroxin and monocyte human leukocyte antigen-DR expression in senior patients with sepsis and explore their clinical significance.

METHODSAccording to the 2001 SCCM/ESICM/ACCP/ATS/SIS international sepsis definitions, 125 senior patients with sepsis free of thyroid conditions were divided into non-severe sepsis group (n=86) and severe sepsis group (n=39), with another 30 healthy subjects as the control. Thyroid function was assayed by chemoluminescence method in these patients and monocyte HLA-DR expression was determined by flow cytometry.

RESULTSCompared with the control group and non-severe sepsis cases, the levels of free T3 (FT3), free T4 (FT4), T3, T4 and monocyte HLA-DR expression were significantly lower in severe sepsis cases (P<0.05), but the levels of thyroid stimulating hormone (TSH) were comparable between the 3 groups (P>0.05). The non-severe sepsis cases showed significantly lower levels of FT3, FT4, T3, T4, TSH and monocyte HLA-DR expression than the control group (P<0.05). In severe sepsis group, the levels of FT3, FT4, T3, T4 and monocyte HLA-DR expression showed significant differences between the fatal cases and surviving cases (P<0.05).

CONCLUSIONThe levels of thyroxin and monocyte human leukocyte antigen-DR expression are obviously lower in senior patients with severe sepsis, and their detection may well indicate the severity of the condition and help make prognostic judgment.

Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; HLA-DR Antigens ; blood ; immunology ; Humans ; Male ; Monocytes ; metabolism ; Pneumonia ; complications ; Sepsis ; blood ; etiology ; immunology ; Thyrotropin ; blood ; Thyroxine ; blood ; Triiodothyronine ; blood

OBJECTIVETo analyze the clinical characteristics of living-related kidney transplantation (LRKT).

METHODSFrom January, 2004 to December, 2008, 175 LRKT were performed including 63 cases (36%) of parent-child relations and 49 cases (28%) of sibling relations between the recipients and donors. Out of 175 donors, 52 were 50 years old or above, 4 had microscopic hematuria (including 2 with also hypertension), 2 had kidney stone, and 2 had high body mass index (BMI). Zero-point graft biopsy was performed in 59 donors, and abnormalities were found in 15 of them. The recipients were at the age of 33-/+10.5 years, and the primary diseases are mainly dominant glomerular nephritis (72.6%, 127/175), and with a few cases of diabetes (4%, 7/175) and hypertensive nephropathy (4%, 7/175).

RESULTSSerum creatinine of the donors was 102-/+22.5 micromol/L at 7 days postoperatively, and 92-/+19.1 micromol/L at one month. One recipient died of severe pulmonary infection. Two recipients underwent graft nephrectomy due to anastomotic stenosis with concomitant acute graft rejection and renal arterial embolism. The one-year survival rates of the patients and grafts were 99.3% and 98.2%, respectively. The incident rates of accelerated rejection and acute rejection were 1.1% and 14.9%, respectively. Other complications included impaired liver function (22.3%), infection (9.7%) and leucopenia (4.6%). The renal arterial stenosis occurred in 2.3% (4/175) of the recipients.

CONCLUSIONSThe recipients of living-related and cadaveric kidney transplant have different primary kidney disease spectrums. Differential diagnosis and treatment of acute rejection and renal artery or anastomotic stenosis can be of vital importance. Marginal donor kidneys with appropriate inclusion criteria can be safely used for transplantation. With good short-term patient and graft survival, LRKT needs further study to evaluate its long-term effect.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Family ; Female ; Glomerulonephritis ; surgery ; Graft Rejection ; epidemiology ; Humans ; Kidney Transplantation ; adverse effects ; Living Donors ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the value of 3D FIESTA sequence in magnetic resonance sialography (MRS) in the diagnosis of obstructive salivary diseases.

METHODSEleven patients with obstructive salivary diseases underwent MRS, and three-dimensional (3D) reconstruction and virtual endoscopic images of the salivary gland ducts were obtained after MRS data post-processing for comparison with those of sialoendoscopy.

RESULTSThe diagnostic accuracy of MRS was 72.7% for obstructive salivary diseases. The virtual endoscopy provided a visual field highly consistent with that by sialoendoscopy.

CONCLUSIONMRS is capable of visualizing the tracts of salivary glands. MR virtual endoscopy can provide sufficient morphological and pathological data for preoperative assessment of salivary operations with sialoendoscopy.

Adult ; Constriction, Pathologic ; diagnosis ; Female ; Humans ; Imaging, Three-Dimensional ; methods ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Parotid Diseases ; diagnosis ; pathology ; Salivary Ducts ; pathology ; Salivary Gland Calculi ; diagnosis ; Salivary Gland Diseases ; diagnosis ; pathology ; Sialography ; methods ; Submandibular Gland Diseases ; diagnosis ; pathology ; Young Adult