LIN28 is an RNA binding protein with important roles in early embryo development, stem cell differentiation/reprogramming, tumorigenesis and metabolism. Previous studies have focused mainly on its role in the cytosol where it interacts with Let-7 microRNA precursors or mRNAs, and few have addressed LIN28's role within the nucleus. Here, we show that LIN28 displays dynamic temporal and spatial expression during murine embryo development. Maternal LIN28 expression drops upon exit from the 2-cell stage, and zygotic LIN28 protein is induced at the forming nucleolus during 4-cell to blastocyst stage development, to become dominantly expressed in the cytosol after implantation. In cultured pluripotent stem cells (PSCs), loss of LIN28 led to nucleolar stress and activation of a 2-cell/4-cell-like transcriptional program characterized by the expression of endogenous retrovirus genes. Mechanistically, LIN28 binds to small nucleolar RNAs and rRNA to maintain nucleolar integrity, and its loss leads to nucleolar phase separation defects, ribosomal stress and activation of P53 which in turn binds to and activates 2C transcription factor Dux. LIN28 also resides in a complex containing the nucleolar factor Nucleolin (NCL) and the transcriptional repressor TRIM28, and LIN28 loss leads to reduced occupancy of the NCL/TRIM28 complex on the Dux and rDNA loci, and thus de-repressed Dux and reduced rRNA expression. Lin28 knockout cells with nucleolar stress are more likely to assume a slowly cycling, translationally inert and anabolically inactive state, which is a part of previously unappreciated 2C-like transcriptional program. These findings elucidate novel roles for nucleolar LIN28 in PSCs, and a new mechanism linking 2C program and nucleolar functions in PSCs and early embryo development.
Animals ; Cell Differentiation ; Embryo, Mammalian/metabolism* ; Embryonic Development ; Mice ; Pluripotent Stem Cells/metabolism* ; RNA, Messenger/genetics* ; RNA, Ribosomal ; RNA-Binding Proteins/metabolism* ; Transcription Factors/metabolism* ; Zygote/metabolism*

OBJECTIVE: To explore the genetic basis for a fetus with cerebellar dysplasia and widened lateral ventricles. METHODS: The couple have elected induced abortion after careful counseling. Skin tissue sample from the abortus and peripheral venous blood samples from both parents were collected for the extraction of genomic DNA, which was then subjected to whole exome sequencing. Candidate variant was verified by Sanger sequencing. RESULTS: Prenatal ultrasonography showed increased nuchal translucency (0.4 cm) and widened lateral ventricles. Magnetic resonance imaging revealed infratentorial brain dysplasia. By DNA sequencing, the fetus was found to carry compound heterozygous variants c.1A>G and c.1564G>A of the RARS2 gene, which were inherited from its father and mother, respectively. Among these, c.1A>G was known to be pathogenic, but the pathogenicity of c.1564G>A was unreported previously. Based on the American College of Medical Genetics and Genomics guidelines, the c.1564G>A variant of RARS2 gene was predicted to be likely pathogenic(PM2+PM3+PP3+PP4). CONCLUSION The compound heterozygous variants c.1A>G and c.1564G>A of RARS2 gene contributed to the fetus suffering from pontocerebellar hypoplasia type 6, which expanded variant spectrum of RARS2 gene.
Female ; Fetus ; Genomics ; Humans ; Mutation ; Olivopontocerebellar Atrophies ; Pregnancy ; Whole Exome Sequencing

Objective:To explore the mechanism of pioglitazone in reducing lung injury induced by acute pancreatitis.Methods:Thirty healthy male SD rats were randomly(random number) divided into the sham operation group, model group and pioglitazone group, with 10 rats in each group. After anesthesia, the rats in the sham operation group were injected with normal saline retrogradely through the pancreaticobiliary duct. In the model group, after anesthesia, the rats were retrogradely injected with sodium taurocholate into the pancreaticobiliary duct to construct the lung injury model of severe acute pancreatitis. In the pioglitazone group, the model was established after intraperitoneal injection of pioglitazone. Six rats in each group were randomly selected and killed 12 h after operation, and then lung tissue and venous blood were collected. The levels of serum amylase and TNF-α and NO in lung tissue homogenate were detected and compared among the three groups; the expression of TLR2 mRNA and TLR4 mRNA in lung tissue was detected by RT-PCR and compared among the three groups; the lung tissue pathological injury score and lung leakage index were calculated and compared among the three groups. The correlation of TLR2 and TLR4’s mRNA expression with lung tissue pathological injury score and lung leakage index was analyzed.Results:The levels of serum amylase and the levels of TNF-α and NO in lung tissue homogenate in the model group were significantly higher than those in the sham operation group, and the above indexes in the pioglitazone group were significantly lower than those in the model group ( P<0.05). The expression levels of TLR2 mRNA and TLR4 mRNA in lung tissue, the lung tissue pathological injury score and lung leakage index in the model group were significantly higher than those in the sham operation group, and the above indexes in the pioglitazone group were significantly lower than those in the model group ( P<0.05). Spearman correlation analysis showed that the expression levels of TLR2 mRNA and TLR4 mRNA in lung tissue were significantly positively correlated with the lung tissue pathological injury score ( rs=0.959, P<0.001; rs=0.924, P<0.001). Pearson correlation analysis showed that the expression levels of TLR2 mRNA and TLR4 mRNA in lung tissue were significantly positively correlated with the lung leakage index ( r=0.957, P<0.001; r=0.958, P<0.001). Conclusions:Pioglitazone may reduce the severity of severe acute pancreatitis induced lung injury by inhibiting the expression of TLR2 mRNA and TLR4 mRNA in lung tissue.

Objective:To explore the mechanism of miR-494 negatively regulating ROCK1 and PTEN in inhibiting apoptosis of pancreatic cells and participating in the occurrence and development of acute pancreatitis.Methods:Pancreatic acinar cells AR42J from rats were treated by caerulein, and then the levels of amylase, tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1) and IL-6 in the supernatant of cell culture were detected by ELISA to verify the cell model of acute pancreatitis. RT-PCR was used to detect the expression of miR-494 in normal AR42J cells (control group) and acute pancreatitis cell model (model group). Flow cytometry was used to detect the apoptosis of the control group, negative control miRNA transfected acute pancreatitis cell model (negative control group) and miR-494 transfected acute pancreatitis cell model (miR-494 transfection group). Western blot was used to detect the expression of ROCK1 and PTEN in the control group, negative control group and miR-494 transfection group.Results:The levels of amylase, TNF-α, IL-1 and IL-6 in the supernatant of AR42J cells treated with caerulein for 8 h and 12 h were significantly higher than those at 0 h and the control group ( P<0.05), indicating that the model was successfully constructed. The expression levels of miR-494 at 8 h, 12 h and 24 h after the establishment of acute pancreatitis cell model were significantly higher than those at 4 h and the control group ( P < 0.05). The apoptosis rate of the model group was significantly higher than that of the control group ( P<0.05), and the apoptosis rate of the miR-494 transfection group was significantly lower than that of the model group ( P<0.05). The expression levels of ROCK1 and PTEN in the miR-494 transfection group were significantly lower than those in the model group and negative control group ( P<0.05). Conclusions:When acute pancreatitis occurs, overexpression of miR-494 can inhibit the expression of pro-apoptotic protein, thus inhibiting the apoptosis of pancreatic acinar cells and promoting the development of acute pancreatitis.

Objective To detect potential variant in a male fetus suspected for Ectrodactyly,Ectodermal dysplasia,Cleft lip/palate (EEC) syndrome.Methods Peripheral blood samples of the fetus and his parents were collected for the extraction of DNA.Whole-exome sequencing was carried out to detect potential variants.Suspected variants were verified by Sanger sequencing.Results The fetus was found to carry a heterozygous c.673C＞T missense variant of the Tp63 gene,which was known to underlie splithand/split-foot malformation.The same variant was not found in either parents.Conclusion The heterozygous c.673C＞ T missense variant of the Tp63 gene probably underlies the EEC syndrome in the fetus.Above finding also expanded the phenotypic spectrum for this variant.

Objective To investigate the effect and application value of HC visual laryngoscope used for the emergency tracheal intubation on obese patients in the Emergency Department. Methods Totally 80 obese patients enrolled from January 2014 to December 2016 from Emergency Department, Second affiliated hospital of Xi'an Jiaotong University who needed the emergency tracheal intubation were randomly (random number) divided into two groups. Patients in group T were operated with traditional laryngoscope, and patients in group HC with HC visual laryngoscope. Then the success rate of glottis exposure, the trial times, operative time, success rate and complication rate of tracheal intubation were compared between the two groups. Results The success rate of glottis exposure in group HC was significantly higher than that in group T (95％ vs 77.5％, P<0.05). The one-time success rate of tracheal intubation and the total success rate of tracheal intubation in group HC were significantly higher than those in group T (72.5％ vs 37.5％, and 95％ vs 62.5％, respectively, P<0.05). However, the trial times of successful intubation cases and the operative time of successful intubation cases were significantly less than those in group T (1.26±0.40) vs (1.64±0.82), and (30.74±6.17) s vs (44.2±7.68) s, respectively, P<0.05. The complication rate of tracheal intubation in group HC was significantly less than that in group T (12.5％ vs 35％, P<0.05). Conclusions HC visual laryngoscope used for the obese patients in Emergency Department can not only increase the success rate of glottis exposure, decrease trial times and shorten operative time of intubation, but also improve the success rate and decrease the complication rate of emergency tracheal intubation, thus having a certain application value.

Objective To discuss the ethical necessity and feasibility of implementing multiple comprehensive psychological intervention measures in intensive care unit (ICU), and to investigate the effect of the measures on treatment compliance. Methods The conscious adult patients hospitalized in emergency ICU (EICU) of the Second Affiliated Hospital of Xi'an Jiaotong University from 2015 to 2017 were enrolled, and divided into control group and intervention group according to random number table, with 40 patients in each group. Patients in control group were treated with routine care and routine diagnosis and treatment, and patients in intervention group were treated with the help of psychologists and with multiple comprehensive psychological intervention measures (including nursing care, medical treatment, family and management) based on routine care and routine diagnosis and treatment. After 5 days of intervention, the treatment compliance, the medical coping attitude and the mental health were assessed respectively using the Treatment Compliance Scale, the Medical Coping Modes Questionnaire (MCMQ) and the Symptom Checklist Scale (SCL-90), and then comparation were done between the two groups. Results After 5 days of intervention, 3 patients in the control group and 4 patients in the intervention group withdraw the study because of the change of illness, and at last 37 patients in the control group and 36 in the intervention group were enrolled. There was no significant difference in basic data such as gender, age, education level, disease, course of disease, the length of EICU stay, and acute physiology and chronic health evaluationⅡ (APACHEⅡ) score between the two groups. The scores of treatment compliance in intervention group was significantly higher than that in control group (2.89±0.67 vs. 2.32±0.91, P < 0.01). Compared with the control group, the "confrontation" dimension score of the MCMQ questionnaire in intervention group was significantly increased (19.75±2.08 vs. 18.62±2.65, P < 0.05), while the "avoidance" and"surrender" dimensions scores were significantly decreased (14.22±1.91 vs. 15.14±1.92, 8.83±1.54 vs. 9.73±2.10, both P < 0.05). In the SCL-90 scale, the scores of the 5 factors such as "somatization", "interpersonal sensitivity","depression", "anxiety" and "photic anxiety" in intervention group were significantly less than those in control group (1.59±0.38 vs. 1.81±0.37, 1.72±0.40 vs. 1.93±0.42, 1.76±0.32 vs. 1.92±0.29, 1.82±0.40 vs. 2.14±0.40, 1.44±0.30 vs. 1.60±0.38, all P < 0.05), while there was no significantly difference in the scores of the other 4 factors as "obsessive-compulsive symptoms", "hostile", "bigotry" and "psychoticism" between the two groups (all P > 0.05). Conclusion The multiple comprehensive psychological intervention measures used for ICU conscious patients could improve the treatment compliance, make the patients' medical coping attitude more active and improve the mental health of the patients, so it has ethical necessity and certain feasibility.

Objective: To explore the role of toll-like receptor 4 (TLR4)/NF-κB signaling pathway in acute necrotizing pancreatitis (ANP)-associated lung injury and the intervention of lipoxin A4 (LXA4) analogue. Methods: Forty-five Sprague-Dawley rats were randomly(random number)divided into the sham operation group, experimental group, and intervention group, each group containing 15 rats. ANP animal models were prepared by injecting sodium taurocholate into biliopancreatic tube in the experimental group. No sodium taurocholate was injected into biliopancreatic duct in the sham operation group. After the preparation of ANP animal models in the intervention group, LXA4 was injected through the tail vein. Rats in each group were randomly divided into 3 subgroups (n=5 each subgroup). The serum amylase, TNF- α, IL-1β, IL-6 and endotoxin levels were detected 6, 12 and 24 h after the operation. The lung injury scores were assessed and the lung wet/dry weight ratio was calculated. The expressions of TLR4 and NF-κB p65 in lung tissues were detected by Western blot. Results: Serum levels of amylase, TNF-α, IL-1β, IL-6 and endotoxin in the experimental and intervention groups were significantly higher than those in the sham operation group, while the levels of the above indicators in the intervention group was significantly lower than those in the experimental group, and the differences were statistically significant (P<0.05). Postoperative lung injury scores and lung wet/dry weight ratio in the experimental group were significantly higher than those in the sham operation group, and the differences were statistically significant (P<0.05). Lung injury scores in the intervention group 6 h after operation had no significant difference compared with those in the sham operation group (P>0.05), while lung wet/dry weight ratio in the intervention group 6 h after operation, and lung injury scores and lung wet/dry weight ratio in the intervention group 12 h or 24 h respectively after operation were significantly higher than those in the sham operation group, with statistically significant differences (P<0.05). Postoperative lung injury scores and lung wet/dry weight ratio in the intervention group were significantly lower than those in the experimental group, and the differences were statistically significant (P<0.05). The expressions of TLR4 and p65 in the lung tissues of the experimental and intervention groups were significantly higher than those of the sham operation group, and the expressions of TLR4 and p65 in the lung tissues of the intervention group were significantly lower than those of the experimental group, with statistically significant differences (P<0.05). Conclusions LXA4 can reduce the severity of acute necrotizing pancreatitis-associated lung injury, and its mechanism is related to reducing the stimulation of endotoxin, thus inhibiting TLR4 signaling pathway and the activation of p65 to down-regulate the level of pro-inflammatory cytokines.

Alzheimer's disease (AD) is a typical neurodegenerative disease, which is clinically manifested as amnesia, loss of language ability and self-care ability, and so on. So far, the cause of the disease has still been unclear and the course of the disease is irreversible, and there has been no cure for the disease yet. Hence, early prognosis of AD is important for the development of new drugs and measures to slow the progression of the disease. Mild cognitive impairment (MCI) is a state between AD and healthy controls (HC). Studies have shown that patients with MCI are more likely to develop AD than those without MCI. Therefore, accurate screening of MCI patients has become one of the research hotspots of early prognosis of AD. With the rapid development of neuroimaging techniques and deep learning, more and more researchers employ deep learning methods to analyze brain neuroimaging images, such as magnetic resonance imaging (MRI), for early prognosis of AD. Hence, in this paper, a three-dimensional multi-slice classifiers ensemble based on convolutional neural network (CNN) and ensemble learning for early prognosis of AD has been proposed. Compared with the CNN classification model based on a single slice, the proposed classifiers ensemble based on multiple two-dimensional slices from three dimensions could use more effective information contained in MRI to improve classification accuracy and stability in a parallel computing mode.
Alzheimer Disease ; diagnosis ; Brain ; diagnostic imaging ; Cognitive Dysfunction ; Deep Learning ; Humans ; Neural Networks (Computer) ; Neuroimaging ; Prognosis