OBJECTIVE: To explore the genetic basis for four patients suspected for Marfan syndrome (MFS). METHODS: Four male patients with suspected MFS and their family members who were treated at West China Second Hospital of Sichuan University from September 12, 2019 to March 27, 2021 were selected as the study subjects. Peripheral venous blood samples were collected from the patients and their parents or other pedigree members for the extraction of genomic DNA. Whole exome sequencing was carried out, and candidate variants were validated by Sanger sequencing. The pathogenicity of the variants was determined based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). RESULTS: Genetic testing revealed that all four patients have harbored variants of the FBN1 gene, including c.430_433del (p.His144fs) deletional variant in exon 5, c.493C>T (p.Arg165*) nonsense variant in exon 6, c.5304_5306del (p.Asp1768del) deletional variant in exon 44 and c.5165C>G (p.Ser1722Cys) missense variant in exon 42. According to the ACMG guidelines, the c.430_433del and c.493C>T were classified as pathogenic variants (PVS1+PM2_Supporting+PP4; PVS1+PS1+PS2+PM2_Supporting+PP4). c.5304_5306del and c.5165C>G were classified as likely pathogenic variants (PS2+PM2_Supporting+PM4+PP4; PS2_Moderate+PS1+PM1+PM2_Supporting). CONCLUSION The c.430_433del and c.5304_5306del variants of the FBN1 gene identified in this study were unreported previously. Above results have enriched the variation spectrum of the FBN1 gene and provided a basis for genetic counseling and prenatal diagnosis of patients with MFS and acromicric dysplasia.
Female ; Pregnancy ; Humans ; Male ; Exons ; China ; Family ; Genetic Counseling ; Genetic Testing ; Marfan Syndrome/genetics* ; Mutation ; Fibrillin-1/genetics*

Objective:To evaluate the role of Jumonji domain-containing 3 (JMJD3) in cisplatin-induced renal fibrosis following acute kidney injury in mice.Methods:Forty-eight healthy C57BL/6 male mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n=12 each) using a random number table method: control group (group CON), control plus JMJD3 inhibitor group (group CON-A), cisplatin group (group CIS), and cisplatin plus JMJD3 inhibitor group (group CIS-A). In group CIS and group CIS-A, cisplatin was intraperitoneally administered on 1st and 14th days, respectively, to develop a renal fibrosis model in mice with acute kidney injury, and the JMJD3 inhibitor GSKJ4 10 mg/kg and equal volume of PBS were intraperitoneally injected on 4th day, respectively, once every 3 days, 6 injections in total.The equal volume of PBS and GSKJ4 10 mg/kg were intraperitoneally injected at the corresponding time points in group CON and group CON-A, respectively.Six mice in each group were selected, and orbital blood samples were collected on 3rd day after the first injection of cisplatin to determine the concentrations of serum creatinine (Cr) and blood urea nitrogen (BUN), then the animals were sacrificed, and kidney tissues were obtained for microscopic examination of pathological changes after HE and PAS staining (with a light microscope), and the damage to kidneys was assessed and scored.Six mice were sacrificed on 28th day after the first injection of cisplatin, and kidney tissues were removed for determination of the area of renal fibrosis ( via Sirius red and Masson staining), expression of fibronectin (Fn), collagen type Ⅰ (Col Ⅰ) and α-smooth muscle actin (α-SMA) (by immunofluorescence), F4/80 + cell and CD3 + cell count (using immunohistochemical method), and expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), CXC chemokine ligand 16 (CXCL16), and monocyte chemoattractant protein1 (MCP-1) mRNA (by real-time polymerase chain reaction). Results:Compared with group CON, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly increased, the expression of Fn, Col Ⅰ and α-SMA was up-regulated, the F4/80 + cell and CD3 + cell count was increased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was up-regulated in group CIS ( P<0.05), and no significant change was found in the parameters mentioned above in group CON-A ( P>0.05). Compared with group CON-A, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly increased, the expression of Fn, Col Ⅰ and α-SMA was up-regulated, the F4/80 + cell and CD3 + cell count was increased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was up-regulated in group CIS-A ( P<0.05). Compared with group CIS, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly decreased, the expression of Fn, Col Ⅰ and α-SMA was down-regulated, the F4/80 + cell and CD3 + cell count was decreased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was down-regulated in group CIS-A ( P<0.05). Conclusions:JMJD3 is involved in the process of renal fibrosis following acute kidney injury in mice, and the mechanism may be related to promotion of inflammatory responses.

Objective:Sepsis patients usually have a fever, but it is still controversial about whether sepsis patients with fever need cooling treatment. This study aimed to evaluate the effect of external physical cooling on the prognosis of sepsis patients.Methods:This study was a single-center, open-label, randomized clinical trial. Adult sepsis patients with body temperature above 38.3 °C admitted to the Critical Care Medicine of Northern Jiangsu People's Hospital from June 2020 to December 2020 were selected, and randomly assigned in a 1∶1 ratio to the cooling group and control group. Patients in the cooling group used external physical cooling methods to reduce their core body temperature to the normal range (36.5-37.5°C) within 4 h of enrollment and maintained for 48 h. Standard care was implemented in the control group at all times, and all antipyretic treatments were prohibited. The 28-day mortality, 72 h-Δ sequential organ failure assessment (SOFA) score (SOFA score at enrollment–SOFA score after 72 h), length of hospital stay and length of ICU stay were compared between the two groups.Results:A total of 53 patients (32 males and 21 females) were enrolled in the study, including 26 patients in the cooling group and 27 patients in the control group. There were no statistical differences in age, sex, source of infection, SOFA score and body temperature between the two groups (all P>0.05). There was no significant difference in the 28-day mortality between the cooling group and the control group ( RR=1.38, 95% CI: 0.62-3.07, P=0.430). The 72 h-ΔSOFA score of the cooling group was significantly higher than that of the control group, the mean difference between the two groups was 1.90 (95% CI: 0.09-3.71, P=0.040), and there was no significant difference in length of hospital stay, length of ICU stay and 28-day mortality between the two groups. Conclusions:External physical cooling management can not significantly reduce the 28-day mortality of sepsis patients. However, external physical cooling can reduce the 72-h SOFA score in sepsis patients, and improve the organ function of patients.

Objective:To evaluate the role of natural killer T cells in renal fibrosis in mice with acute kidney injury (AKI).Methods:Twenty-four clean-grade healthy male C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), AKI group (group A), control plus CD1d antibody group (group C-MA), and AKI plus CD1d antibody group (group A-MA). The model of renal fibrosis in mice with AKI was established by intraperitoneal injection of folic acid 250 mg/kg.In group C, homotypic control antibody 20 mg/kg was injected via the tail vein.In group AKI, homotypic control antibody 20 mg/kg was injected via the tail vein at 24 h before establishing the model. In group C-MA, anti-CD1d monoclonal antibody 20 mg/kg was injected via the tail vein.In group A-MA, anti-CD1d monoclonal antibody 20 mg/kg was injected via the tail vein at 24 h before establishing the model.On the 14th day after folic acid injection, blood samples were taken from eyeballs to determine the concentrations of blood urea nitrogen (BUN) and creatinine (Cr) in serum.Then the mice were sacrificed, and the renal tissues were taken for Sirius red staining and HE staining to determine the area of renal fibrosis, and renal injury was scored.The expression of fibronectin (FN), type I collagen (Col-Ⅰ) and alpha-smooth muscle actin (α-SMA) in renal tissues was detected by immunofluorescence method.The expression of interleukin (IL)-4, IL-13, arginase-1 (Arg-1) and found in inflammatory zone 1 (FIZZ1) mRNA in renal tissues was detected by real-time polymerase chain reaction. Results:Compared with group C, the concentrations of BUN and Cr in serum, renal injury score, and area of renal fibrosis were significantly increased, the expression of FN, Col-Ⅰ and α-SMA and IL-4, IL-13, Arg-1 and FIZZ1 mRNA was up-regulated in A and A-MA groups ( P<0.05), and no significant change was found in the above indexes in group C-MA ( P>0.05). Compared with group A, the concentrations of BUN and Cr in serum, renal injury score, and area of renal fibrosis were significantly decreased, the expression of FN, Col-Ⅰ and α-SMA and IL-4, IL-13, Arg-1 and FIZZ1 mRNA was down-regulated in group A-MA ( P<0.05). Conclusion:Activation of natural killer T cells is involved in the process of renal fibrosis in mice with AKI, and the mechanism may be related to promoting the release of Th2 cytokines and M2 polarization of macrophages.

The secondary structure is a fundamental feature of both non-coding RNAs (ncRNAs) and messenger RNAs (mRNAs). However, our understanding of the secondary structures of mRNAs, especially those of the coding regions, remains elusive, likely due to translation and the lack of RNA-binding proteins that sustain the consensus structure like those binding to ncRNAs. Indeed, mRNAs have recently been found to adopt diverse alternative structures, but the overall functional significance remains untested. We hereby approach this problem by estimating the folding specificity, i.e., the probability that a fragment of an mRNA folds back to the same partner once refolded. We show that the folding specificity of mRNAs is lower than that of ncRNAs and exhibits moderate evolutionary conservation. Notably, we find that specific rather than alternative folding is likely evolutionarily adaptive since specific folding is frequently associated with func-tionally important genes or sites within a gene. Additional analysis in combination with ribosome density suggests the ability to modulate ribosome movement as one potential functional advantage provided by specific folding. Our findings reveal a novel facet of the RNA structurome with important functional and evolutionary implications and indicate a potential method for distinguishing the mRNA secondary structures maintained by natural selection from molecular noise.

The dynamic coupling of stent degradation and vessel remodeling can influence not only the structural morphology and material property of stent and vessel, but also the development of in-stent restenosis. The research achievements of biomechanical modelling and analysis of stent degradation and vessel remodeling were reviewed; several noteworthy research perspectives were addressed, a stent-vessel coupling model was developed based on stent damage function and vessel growth function, and then concepts of matching ratio and risk factor were established so as to evaluate the treatment effect of stent intervention, which may lay the scientific foundation for the structure design, mechanical analysis and clinical application of biodegradable stent.
Biomechanical Phenomena ; Constriction, Pathologic ; Humans ; Stents

Objective: To evaluate the diagnostic efficacy of Japan Narrow Band Imaging Expert Team(JNET) classification under narrow-band imaging (NBI) for colorectal laterally spreading tumors. Methods: Data of 170 laterally spreading tumors (LST) detected by NBI and pigment dyeing were reviewed in the retrospective study. JNET classification under NBI was used for rediagnosis based on surface pattern and vessel pattern. Pit pattern(PP) was observed under pigment dyeing using PP classification. The results were compared with histologic results after endoscopic resection or surgery. Results: The diagnostic sensitivity, specificity, positive predictive value, negative predictive value and accuracy of JNET classification and PP classification were 92.2% VS 70.3%, 82.3% VS 85.0%, 74.7% VS 72.6%, 94.9% VS 83.5%, 85.9% VS 79.7%, respectively (P=0.159). The consistency rates of JNET classification and PP classification in predicting shallow invasion depth of LST were 6.1% and 8.3% respectively and the consistency rates in predicting deep invasion were 30.8% and 4.8%, respectively. Conclusion JNET classification under NBI is effective in predicting malignant laterally spreading tumors, however, its efficacy in predicting tumor invasion depth is unsatisfied. PP classification can be used to improve the diagnostic accuracy for those with diagnostic difficulty.

Objective To evaluate the role of phosphatidylinositol 3-kinase/serine-threonine kinase(PI3K/Akt) signaling pathway in mitochondrial fission in endotoxin-challenged alveolar type Ⅱ epithelial cells of rats.Methods Rat alveolar type Ⅱ epithelial cells CCL-149 were seeded in 6-well plates at a density of 2×105 cells/ml.CCL-149 cells were divided into 6 groups (n =10 each) using a random number table:control group (group C),lipopolysaccharide (LPS) group (group L),LPS plus CO-releasing molecule-2 (CORM-2) group (group L+CO),LPS plus PI3K inhibitor LY294002 group (group L+LY),LPS plus iCORM-2 group (group L+iCO) and LPS plus dimethyl sulfoxide (DMSO) group (group L+D).CCL149 cells were stimulated with 10 μg/ml LPS for 24 h in L,L+CO,L+LY,L+iCO and L+D groups.CORM-2 100 μmol,LY294002 25 μg and iCORM-2 100 μmol were added at 1 h before stimulation with LPS in L+CO,L+LY and LPS+iC0 groups,respectively.In group L+D,0.1％ DMSO 100 μmol was added at 1 h before stimulation with LPS.After the end of incubation,the concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the culture medium were determined by enzyme-linked immunosorbent assay,and the expression of phosphorylated-Akt (p-Akt),heme oxygenase-1 (HO-1),dynamin-related protein 1 (Drpl) and fissionl (Fisl) was detected by Western blot.Results Compared with group C,IL-6 and TNF-α concentrations in the culture medium were significantly increased,and the expression ofp-Akt,HO-1,Drp1and FIS1 was up-regulated in L,L+CO,L+LY,L+iCO and L+D groups (P＜0.05).Compared with group L,IL-6 and TNF-α concentrations in the culture medium were significantly decreased,the expression of p-Akt and HO-1 was up-regulated,and the expression of Drp1 and Fis1 was down-regulated in group L+CO,and IL-6 and TNF-α concentrations in the culture medium were significantly increased,the expression of p-Akt and HO-l was down-regulated,and the expression of Drpl and Fisl was up-regulated in group L+LY (P＜0.05).Conclusion Activation of PI3K/Akt signaling pathway can inhibit mitochondrial fission in endotoxin-challenged alveolar type Ⅱ epithelial cells of rats.

Background:DNA methylation is a vital part of epigenetic modification,and is closely related with the development and progress of multiple tumors such as colorectal cancer. However,its mechanism is not fully clarified. Screening specific methylation gene and construct the methylation expression profile of tumor has become the current research hotspot. Aims:To screen the differential methylation loci in colorectal cancer and para-cancerous normal tissue by gene methylation microarray technique,and to construct specific differential methylation gene profile of colorectal cancer. Methods:Methylation 450K bead-chip was applied to detect the methylation status in colorectal cancer and para-cancerous normal tissues of 6 cases. A total of 431 467 loci were analyzed and compared. Aberrant methylation loci were screened according to P value,and the hypermethylation loci and hypomethylation loci were differentiated by delta beta value. Moreover,the function of differential methylation gene was further analyzed by GO analysis and KEGG analysis. Results:A total of 3 649 differential methylation loci were found by comparing colorectal cancer tissue and para-cancerous normal tissue,including 1 259 hypermethylation loci,which mainly located in promoter and genosome,and 2 390 hypomethylation loci,which mainly located in intergenic region and genosome. A panel of aberrant methylation gene loci was screened out,including hypermethylation gene loci such as SLC15A3 and hypomethylation gene loci such as ACOT2,TTLL8 and UHRF1. GO analysis and KEGG analysis showed that the function of these genes might be correlated with DNA binding,transcription factor activity and signal transduction pathway. Conclusions:There are many differential methylation loci in colorectal cancer and para-cancerous normal tissues,suggesting that aberrant DNA methylation is closely related with the development and progress of colorectal cancer. DNA methylation microarray technique could be applied for preliminary screening of differential methylation loci. However,constructing the differential methylation profile in colorectal cancer as a clinical biomarker should be further validated.

Non-coding RNA (ncRNA) is a class of RNA not coding the protein and plays a significant role in the process of growth and development of diseases.Barrett''s esophagus (BE) is considered as the precancerous lesion of esophageal adenocarcinoma.Several studies showed that ncRNA has the potential value in diagnosis, treatment and designing of targeting drugs.This article reviewed the role of ncRNA, especially microRNA, long non-coding RNA and circular RNA in the development and carcinogenic process of BE.