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Objective:To investigate the clinical and genetic mutation characteristics of a pedigree with familial exudative vitreoretinopathy (FEVR) associated with a LRP5 gene mutation. Methods:A pedigree investigation was performed in a two-generation Chinese Han family with FEVR, which was diagnosed in The First Affiliated Hospital of Xi'an Jiaotong University.Three family members, the proband and his parents, underwent ophthalmic examination, including visual acuity, intraocular pressure, slit-lamp microscopy, fundoscopy and wild-field fundus fluorescein angiography (FFA), to clinically characterize the FEVR phenotype.Peripheral blood of the families were collected for high-throughput sequencing and bioinformatics analysis to identify the pathogenic gene.Sanger sequencing verification was conducted for the detected mutation.The pathogenicity of identified mutations was analyzed according to the guidelines of the American Association of Medical Genetics (ACMG) with software such as Mutation Taster, Polyphen-2, PROVEN and REVEL.This study adhered to the Declaration of Helsinki and was approved by an Ethics Committee of The First Affiliated Hospital of Xi'an Jiaotong University (No.2017-740). Written informed consent was obtained from each subject.Results:The proband was a 27-year-old male.Uncorrected visual acuity (UCVA) was 1.0 for his right eye and 1.2 for his left eye.Fundoscopy showed vascular tortuosity and vasodilation in the temporal peripheral retina of both eyes.FFA indicated that hairbrush-like vasculature and non-perfusion lesion in peripheral retina.The best-corrected visual acuity (BCVA) of the proband's mother, a 51-year old female, was 1.0 for the both eyes.Temporal retinal vascular tortuosity in her left eye was found with fluorescein leakage detected by FFA.The BCVA of the proband's father, a 56-year-old male, was 1.0 for both eyes.Leopard fundus and optic disc atrophy were visible.FFA result was normal.The result of genetic test showed that there were two novel gene mutations in the family with LRP5 gene c. 4110T>G(p.Cys1370Trp) and FSCN2 gene c. 1495G>A(p.Gly499Ser). According to guidelines of ACMG, LPR5 c. 4110T>G was a mutation with uncertain significance.Multiple software, such as MutationTaster, Polyphen-2, PROVEN and REVEL predicted that LRP5 c. 4110T>G mutation might cause detrimental effects on genes or gene products.REVEL scale was 0.93, which indicated that it might be a pathogenic variant. Conclusions:LRP5 gene c. 4110T>G(p.Cys1370Trp) may be a novel mutation for FEVR, which enriches the mutation spectrum of LRP5 gene.
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Alternative splicing is a tightly regulated process that contributes to cancer development. CRNDE is a long noncoding RNA with alternative splicing and is implicated in the pathogenesis of several cancers. However, whether deregulated expression of CRNDE is common and which isoforms are mainly involved in cancers remain unclear. In this study, we report that CRNDE is aberrantly expressed in the majority of solid and hematopoietic malignancies. The investigation of CRNDE expression in normal samples revealed that CRNDE was expressed in a tissue- and cell-specific manner. Further comparison of CRNDE expression in 2938 patient samples from 15 solid and hematopoietic tumors showed that CRNDE was significantly overexpressed in 11 malignancies, including 3 reported and 8 unreported, and also implicated that the overexpressed isoforms differed in various cancer types. Furthermore, anti-cancer drugs could efficiently repress CRNDE overexpression in cancer cell lines and primary samples, and even had different impacts on the expression of CRNDE isoforms. Finally, experimental profiles of 12 alternatively spliced isoforms demonstrated that the spliced variant CRNDE-g was the most highly expressed isoform in multiple cancer types. Collectively, our results emphasize the cancer-associated feature of CRNDE and its spliced isoforms, and may provide promising targets for cancer diagnosis and therapy.
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[Objective]The oxidative injury of retinal pigment epithelium(RPE)plays a key role in the pathogenesis of age-related macular degeneration(ARMD). This study is to investigate the effects of endoplasmic reticulum stress and the vital transcriptional factor X-box binding protein 1(XBP1)in acrolein-induced oxidative damage of RPE.[Methods]RPE cells were treated with acrolein (75μmol/L)for 2~24 h,expression of glucose regulated protein 78(GRP78)and XBP1 was determined by Western blot analysis. After being transfected with XBP1 siRNA with 24 h,the expression of XBP1 was knocked-down in RPE cells. Protein level of Nrf2 and SOD2 was then determined by Western blot analysis and intracellular Reactive Oxygen Species(ROS)generation was determined by DCF staining. Acrolein was added for 8 h after being transfected with XBP1 siRNA or control siRNA for 24 h. Apoptosis was detected by TUNEL assay before and after the treatment. Subretinal injection of Cre or GFP adenovirus was performed in XBP 1flox mice. After 1 week the mice were sacrificed. Total RNA was extracted from mice eyecups using TRIzol and real-time RT-PCR was performed to determine the two XBP1 down-stream genes,ERdj4 and p58IPK. Cryosectioning and immunofluorescent staining were performed to look at the expression of XBP1,Nrf2 and SOD2 in mice RPE.[Results]Protein level of GRP78 was significantly un-regulated after exposure to acrolein for 2 and 4 h. XBP1 was activated after acrolein treatment for 6 h. Knock-down of XBP1 by siRNA down-regu?lates anti-oxidant genes expression and increased ROS generation in RPE cells. Loss of XBP1 exacerbates acrolein-induced cell apoptosis. XBP1 was knocked-down in the RPE of XBP1flox mice after subretinal injection of Cre adenovirus. Decreased mRNA level of ERdj4 and p58IPK,and decreased Nrf2 and SOD2 expression were seen in the Cre-injected group.[Conclusions]Acrolein induces ER stress and activates XBP1 in RPE cells. Knock-down of XBP1 down-regulates anti-oxidant genes expression ,increases ROS generation,and exacerbates acrolein-induced cell apoptosis. XBP1 plays a role in the anti-oxidant defense in the RPE cells.
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Objective To analyze the immune privilege of lung in acute graft-versus-host disease (aGVHD).Methods The models of aGVHD were established,and C57BL/6J→C57BL/6J model was used as control.The clinical scores and survival were observe& The pathological injuries were compared between the lung and traditional target organs (liver,small intestines and skin).The expression of IFN-γ in different organs after transplantation was detected by ELISA.Results Allogeneic hematopoietic stem cell transplantation (HSCT) mice had high 42-day survival rate (20 ％) post-transplantation,and recipients of allogeneic grafts showed classical symptoms and histological injury,and pathological changes of lung were not as serious as the liver,small intestine,skin at day 28 after transplantation.Syngeneic mice all survived at day 42 after transplantation,without GVHD symptoms and pathological changes.The mice in MHC-disparate mice (2 ∶ 1 and 4 ∶ 1 groups) died significantly faster at a median of 12 days after transplantation,with severe changes of clinical symptoms and pathology of classical organs,and MHC-disparate mice (4∶1 groups) had developed severe interstitial pneumonitis.The mean IFN-γ concentration in the lung of allogeneic HSCT mice was obviously increased in the first and second week after transplantation,the IFN-γ concentrations of target organs (liver,small intestines,skin) were slightly increased in the first and second week after transplantation,and there were statistically significant difference from lung (P＜0.05).Condmion There wasrelativeimmune privilege of lung in a murine model of aGVHD induced by HSCT,which was associated with the expression of MHCin the mice and IFN-gamma of lungs after transplantation.
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OBJECTIVE To analyse the infection of Acinetobacter baumannii(ABA) in ICU of Cardiac Surgery,and make analysis for its resistance to 14 antibiotics.METHODS The collection of clinic stains of ABA isolated from ICU in 2008.The pathogens were identified by VITEK-Jr.RESULTS The drug resistance rates of Imipenem and cefoperazone/sulbactam were 74.4% and 18.6%.The resistance rates to other drugs were more than 70%.CONCLUSIONS ABA strains isolated show the trend of multi-durg resistance.We should pay more attention to the monitor of the resistance to antibiotics.
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Objective To analyze the clinical features,and prognosis of the interstitial lung disease (ILD) in patients with dermatomyositis (DM) and polymyositis (PM) by chest X-ray,chest high-resolution CT scan (HRCT) and pulmonary lung function.Methods Thirty-three patients hospitalized with DM/PM associated ILD were retrospectively analyzed.Results Thirty-three patients with ILD were confirmed by HRCT.Abnormal pulmonary function tests were available in 82% of patients.Clinical-imaging analysis revealed that the pathological features of ILD were non-specific interstitial pneumonia (NSIP,57%) and unusual interstitial pneumonia (UIP,25%).UIP types showed a poor prognosis and high mortality (70%).Conclusion This study shows that HRCT is more sensitive for the diagnosis of ILD than lung function tests and chest X-ray.Combined HRCT and chest X-ray with lung function tests and blood gas analysis have shown that the major pathological types of ILD are NSIP and UIP,in which UIP are associated with high mortality and poor prognosis.
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To construct a pUCP18/lasRantisense plasmid carrying the reversed gene and analyze its effect on the virulence of Pseudomonas aeruginosus, LasR gene was amplified from the genome of Pseudomonas aeruginosus by PCR and reversely recombined with plasmid pUCP18. The recombinant pUCP18/lasRantisense was verified by enzyme digestion, PCR and sequencing. The biological effects of pUCP18/lasRantisense were examined by using RT-PCR, NAD method and the assay of pyocyanin. Our results showed that the expected full length lasR fragment (721 bp) was extended from Pseudomonas aeruginosus gene with PCR. And it is consistent with LasR gene of Pseudomonas aeruginosa in GenBank (No. NC_002516). The recombinant plasmid was successfully constructed and transferred into Pseudomonas aeruginosus. The antisense nucleic acid of LasR gene could reduce the virulence of Pseudomonas aeruginosus and might serve as a new target site for treatment purpose.
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The LasR gene was amplified from the genome of Pseudomonas aeruginosus by PCR and recombined with plasmid pUCP18 reversely. The recombinant pUCP18/lasRantisense was verified with restriction analysis, PCR and sequence and was transformed in Pseudomonas aeruginosus. The biological effect of pUCP18/lasRantisense was detected by RT-PCR, NAD method and the assay of pyocyanin. The air tubes of rats were infected by pUCP18/lasRantisense strain and then carried on histopathologic slide check. Expected full length LasR fragment (721bp) can be extended from Pseudomonas aeruginosus gene with PCR technology. And it is consistent with LasR gene of Pseudomonas aeruginosa covered in GenBank (NO. NC_002516). The recombinant plasmid was constructed and transformed into Pseudomonas aeruginosus sucessfully. Compared with the rats which were infected by standard strain, the bronchitis of the rats which were infected by pUCP18/lasRantisense strain was obviously eased. It can be concluded that the antisensenucleic acid of LasR gene can depress the virulence of Pseudomonas aeruginosus and reveal a new target site for treatment.
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AIM To Investigate the effect of breviscapine on delayed outward potassium current in single ventricular cell of guinea pig. METHOD Using whole cell patch clamp technique. RESULTS Breviscapine enhanced I k dose dependently. Breviscapine with the concentration of 0^01 and 0^02 mg?L -1 increased I k by 44.6% and 78.3%,respectively. The effect could be reversed by washing. CONCLUSION Breviscapine can open the K + channel and incrase I k .