Objective To identify and observe the pathogenic gene variant and clinical phenotype in a family with Leber congenital amaurosis (LCA). MethodsA retrospective clinical study. Two patients and four family members from one LCA family (type 7), diagnosed via genetic testing at the First Affiliated Hospital of Xi'an Jiaotong University in January 2024 were included. Detailed patient and family histories were collected. All patients underwent examinations including best-corrected visual acuity (BCVA), intraocular pressure, color fundus photography, fundus autofluorescence (FAF), flash visual evoked potential (F-VEP), full-field electroretinography (ff-ERG), and optical coherence tomography (OCT). Family members underwent BCVA and color fundus photography examinations. Peripheral venous blood (5 ml) was collected from the patients and the four family members for genomic DNA extraction. High-throughput sequencing was used to screen for pathogenic gene variants. Identified variants were verified by Sanger sequencing. All variants were classified according to the American College of Medical Genetics and Genomics (ACMG) guidelines. Bioinformatics software including Mutation Taster, Polyphen-2, PROVEAN, and REVEL was used to analyze the pathogenicity of the variants. Results The proband (Ⅱ-2), a 14-year-old female, was born to consanguineous parents (first cousins). Her BCVA was 0.1 in both eyes; intraocular pressure was normal; the anterior segments showed no significant abnormalities. Color fundus photography showed waxy optic discs and a "coin-shaped," "salt-and-pepper" appearance in the retina. FAF revealed large areas of hypoautofluorescence in the macular region. OCT showed shallowing or disappearance of the foveal, disorganized retinal layers, and absence of the ellipsoid zone. F-VEP showed recordable P2 waves with no significant delay in peak time but slightly reduced amplitude. ff-ERG showed significantly reduced or non-detectable amplitudes of the scotopic and photopic a- and b-waves. The proband's elder sister (Ⅱ-1) had similar BCVA and fundus findings. The proband's parents (Ⅰ-1, Ⅰ-2), younger brother (Ⅱ-3), and younger sister (Ⅱ-4) showed no significant ocular phenotypic abnormalities. Genetic testing revealed that the proband and her elder sister were homozygous for the CRX gene variant c.122G>A:p.Arg41Gln. The proband's father, mother, and younger brother were heterozygous carriers of the same CRX variant; the younger sister showed no variation at this locus. Based on the clinical presentation, ff-ERG, and genetic test results, the final diagnosis was LCA type 7. According to ACMG guidelines, the c.122G>A variant was classified as likely pathogenic. Mutation Taster and Polyphen-2 software predicted the variant to be damaging; the REVEL score was 0.929, indicating a likely pathogenic variant. Conclusions The homozygous CRX gene variant c.122G>A:p.Arg41Gln causes autosomal recessive LCA type 7 in this family. LCA is characterized by early onset and severe visual impairment.
Objective To detect gene mutations of Fas gene death domain （exons 7-9） in 2 Chinese keloid pedigrees and to investigatethe significance of Fas gene mutations in the keloid formation.Methods The samples were selected from keloid pedigrees A and B in 2005. The polymerase chainreaction and DNA sequencing analysis technique were used to detect the sequenceof exons 7-9 of Fas gene from keloid tissues of 2 male patients in pedigree A,their peripheral vein blood and their surrounding normal skin served as their own contrast, their spouses’ peripheral vein blood served as normal contrast, the peripheral vein blood of 2 patients in pedigree B served as a contrast between different keloid pedigrees.Results No gene mutations and single nucleotidepolymorphism in Fas gene exons 7, 8 were found in all samples from pedigrees A and B. But point mutations and single nucleotide polymorphism in Fas gene exon 9were identified in 11 bp and 53 bpin 2 keloid tissue samples from Chinese keloid pedigree A.Conclusion Fas gene point mutations maybe indicate some relations in Fas protein function and keloid formation.
Genetic epilepsy with febrile seizures plus (GEFS+) is a new type of genetic epilepsy syndrome with a marked hereditary tendency. Febrile seizure is the most common clinical symptom, followed by febrile seizure plus, and with/without absence seizures, focal seizures, and generalized tonic-clonic seizures. Results of the polymerase chain reaction (PCR), exon sequencing and single nucleotide polymorphism (SNP) analysis showed that the occurrence of GEFS+ is mainly related to the mutation of gamma aminobutyric acid type A receptor gamma 2 subunit (GABRG2), but its pathogenesis was still unclear. The main types of GABRG2 mutations include missense mutation, nonsense mutation, frameshift mutation, point mutation and splice site mutation. All these types of mutations can reduce the function of ion channels on cell membrane, but the degree and mechanism of dysfunction are different, which may be the main mechanism of epilepsy. This article will focus on the relationship between GEFS+ and the mutation types of GABRG2 in recent years, which is of great significance for clinical accurate diagnosis, anti-epileptic treatment strategy and new drug development.
ObjectiveTo explore the relationship of the clinicopathological characteristics of non-small cell lung cancer (NSCLC) with the mutations of epidermal growth factor receptor (EGFR), K-ras and EML4-ALK fusion gene in cell blocks of pleural effusion (PLE). MethodsA total of 268 cytological specimens of PLE (pleural effusion), from Central Hospital of Zibo city were collected from advanced NSCLC patients between January 2012 year and June 2014 year. There were 165 male and 103 female patients at age of 53.6 (31-76) years. Qualitative diagnosis has been made in the 268 patients using PLE samples with conventional smear. Immunohistochemical staining combined with cell block section were used for further classification. There were 76 patients diagnosed as NSCLC with 39 patients of adenocarcinoma and 37 patients of squamous-cell carcinoma. In the 76 patients of lung biopsy specimens and PLE, EGFR and K-ras mutations, EML4-ALK fusions were tested. ResultsEGFR mutations rate was 34.21% (26/76). K-ras mutations rate was 6.58% (5/76). EML4-ALK fusions rate was 7.89% (6/76) at the same time. EGFR and K-ras mutations, EML4-ALK fusions were mostly found in young female adenocarcinoma patients who were non-smokers. EGFR and K-ras mutations or EML4-ALK fusions were not found in the same patient. ConclusionCytological specimens are feasible for detecting EGFR were K-ras mutations and EML4-ALK fusions. This will especially benefit to patients whose histological specimen can not be obtained.
Systemic therapy is the main treatment for advanced non-small cell lung cancer, but the effect of chemotherapy alone is not good. In recent years, with the discovery of the pathogenic targets of non-small cell lung cancer, new treatment methods such as targeted drugs and immune checkpoint inhibitors are available, which greatly improve the survival time and quality of life of patients with advanced non-small cell lung cancer. Genetic testing is recommended for all patients with advanced non-small cells lung cancer to obtain more precise and individualized treatment. This article focuses on different types of gene mutations and the corresponding molecular targeted drugs in advanced non-small cell lung cancer, in order to better guide clinical treatment.
Objective To systematically review the association between prothrombin gene G20210A mutation and the risk of cerebral venous thrombosis (CVT). Methods Databases including PubMed, Springer, Google Scholar, The Cochrane Library (Issue 1, 2016), CNKI, WanFang Data and CBM were searched for case-control studies concerning the association between prothrombin gene G20210A mutation and cerebral venous thrombosis risk from inception to January 2016. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Then, meta-analysis was performed using RevMan 5.3 software and Stata 12.0 software. Results A total of 26 case-control studies were included, involving 1 361 CVT cases and 6 323 controls. The results of meta-analysis showed that: there was a significant association between prothrombin gene G20210A mutation and CVT risk (OR=4.56, 95% CI 3.51 to 5.93,P<0.000 01). Sensitivity analysis showed no significant publication bias was detected confirmed the stability of results. Subgroup analysis showed that G20210A mutation increased CVT risk in adults (OR=5.02, 95% CI 3.81 to 6.60,P<0.000 01), but not in children (OR=1.99, 95% CI 0.83 to 4.79,P=0.12). Conclusion Prothrombin gene G20210A mutation can significantly increase the CVT risk. Due to the limited quality and quantity of included studies, the above results are needed to be validated by more high quality studies.
Objective To research the clinical characteristics and the arysulfatase A(ARSA) gene screening inafamily withametachromatic leukodystrophy and epilepsy child. Methods Clinical data were collected and ARSA gene were tested by PCR and Sanger sequencing in the pedigree． Results Two mutations in exon 2 of ARSA gene was identified in the proband includingaknown heterozygous missense mutation c.293C>T which was also found in his mother andanovel frameshift mutation c.302de1G. None of them was found in the proband’s brother. Conclusion The intractable epilepsy of the proband was related to his metachromatic leukodystrophy. Andanew frameshift mutation c.302delG was found in his ARSA gene, which haven’t reported around the world yet. Combined with the patient’s typical late infantile presentation, we speculated that the frameshift mutation c.302delG may be the cause of MLD.
Objective To study the mutations at 1 573 fragment of TNF receptor II (TNFR-II) gene in patients with keloid. Methods The tissue DNA was extracted from 22 samples of keloids donated by 22 patients (6 males and 16 females, aged 18-53 years), and all keloids were examined and classified by pathologist. The peri pheral blood DNA was extracted from the same patients as the control. PCR was used to ampl ify the 1 573 fragment of TNFR-II gene from the keloid tissue DNA and peripheral blood DNA. The PCR products were sequenced directly and then compared with the GeneBankdata. Results All the concentration of the extracted DNA in trial were higher than 0.50 μg/μL and the purity (A260/A280) ofthe extracted DNA were higher than 1.5. It closed to the magnitude of the design DNA fragment by agarose gel electrophoresis examining, and corresponded with the test requirement. Mutations at 1 573 fragment of TNFR-II gene were detected in 13 out of 22 keloids. The mutation incidence was 59.1%. Among them, 9 had point mutation at codon 1 663, accounting 40.9%. No TNFR-II gene mutation was detected in all peripheral blood samples. There were significant difference between keloids DNA and peripheral blood DNA (P lt;0.01). The mutations involved point mutation, deletion and insertion as well as multisite and multitype. Conclusion There is a correlation between the mutation at 1 573 fragment of TNFR-II gene and keloid.
Objective To investigate the feature of c-kit gene mutation in gastrointestinal stromal tumor （GIST） and its correlation with clinicolpathology， molecular targeted therapy，and prognosis. Methods The related literatures about the molecular genetic mechanism of GIST were reviewed. Results The c-kit gene mutation， which is prevalent in GIST， may be the early genomic events， and they are not the independent prognostic factor. However， different molecular subtype as a new indicator to regulate biological behaviors and assess prognosis of GIST is still controversial. Conclusions The study of genotype in GIST has advanced our understanding of pathogenesis， evaluating the prognosis and conducting treatment optimization. However， subsequent work remains to be done.
ObjectiveTo explore the action of dominant-negative effect on mutant insulin gene-induced diabetes.Methods293T cells were transfected with a recombinant plasmid containing mutant preproinsulinogen complementary DNA (cDNA) and a recombinant plasmid containing human wild-type preproinsulinogen cDNA. There were 5 mutant groups which mutant preproinsulins respectively bear substitutions V(A3)L, C(A7)Y, R(SP6)H, G(B8)S or G(C28)R. Wild-type mouse preproinsulin and wild-type human preproinsulin were co-transfected as normal control group. After 48 hours, medium and cells were collected. Human proinsulin were detected by human-specific proinsulin radioimmunoassay.ResultsCompared with the control group [(135.84±1.89) pmol/L], human proinsulin levels in medium of C(A7)Y group [(29.28±6.85) pmol/L] and G(B8)S group[(33.62±10.52) pmol/L] decreased significantly (P<0.01). There was no significant difference in human proinsulin level between the other groups and the control group (P>0.05).ConclusionMutants C(A7)Y and G(B8)S induce the dominant-negative effect on co-existing wild-type proinsulin.