Analysis of clinical phenotype and gene variants in a patient with classic tuberous sclerosis complex / 中华皮肤科杂志

Objective:To analyze clinical phenotypes and pathogenic mutations of a patient with classic tuberous sclerosis complex.Methods:Clinical data was collected from a patient with classic tuberous sclerosis complex. Next-generation sequencing was performed to screen pathogenic gene variants, and Sanger sequencing to verify the mutations. Minigene plasmids were constructed and transfected into the human renal epithelial cell line 293T, and RNA was extracted for transcriptional analysis.Results:The patient clinically presented with recurrent epileptic seizures, facial angiofibroma, periungual fibroma, pulmonary lymphangioleiomyomatosis, renal angiomyolipoma and multiple osteosclerosis. Next-generation sequencing revealed a suspected pathogenic variant in the TSC2 gene in the patient. Sanger sequencing identified a heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene in the patient, but not in his parents or 100 unrelated healthy controls. Moreover, this mutation had not been previously reported. The minigene experiment showed changed mRNA sequence of the TSC2 gene in this patient with loss of the authentic splice site in exon 4 and insertion of a 74-bp intron, which shifted the splice site 90 bp downstream (r.336delins336+16_336+90) .Conclusion:The novel heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene can lead to aberrant splicing, and may contribute to tuberous sclerosis complex in this patient.

Study on the Spectrum-effect Relationship of Antioxidant Activity of Ethanol Extract from Melastoma dodecandrum / 中国药房

OBJECTIVE： To establish the fingerprint of the ethanol e xtract from Melastoma dodecandrum ， to study spectrum-effect relationship of its antioxidant activity. METHODS ：HPLC method and Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）were used to establish the fingerprints of 15 batches of ethanol extracts from M. dodecandrum，and heir similarity was evaluated. The common peaks were identified by comparing with substance control. DPPH free radical scavenging method ，ABTS free radical scavenging method and total antioxidant capacity determination method （FRAP） were used to determine antioxidant activity in vitro of 15 batches of ethanol extracts from M. dodecandrum . Principal component analysis，bivariate correlation analysis and partial least squares regression analysis were used to study the spectrum-effect relationship of the antioxidant activity of the ethanol extracts from M. dodecandrum . RESULTS ：Totally 20 common peaks were identified in HPLC fingerprints of 15 batches of ethanol extracts from M. dodecandrum ；its similarity with the control fingerprint was not less than 0.831；it was identified that peak 3 was gallic acid ，peak 13 was vitexin ，peak 17 was rutin and peak 19 was ellagic acid. The IC 50 values of DPPH radical scavenging method of 15 batches of ethanol extracts from M. dodecandrum were 21.98-57.87 μ g/mL，that of ABTS radical scavenging method were 40.94-101.88 μ g/mL，the results of FRAP method were 0.19-0.48 mg/mL. Principal component analysis showed that the contribution rate of IC 50 variance of DPPH was 80.77％. Bivariate correlation analysis showed that the peak areas of peak 2 （positive correlation ） and peak 11 （negative correlation ） were significantly correlated with antioxidant activity （P＜0.05）；partial least squares regression analysis showed that ，the variable projection importance （VIP）in descending order was peak 11＞peak 2＞peak 16＞peak 15＞peak 12＞peak 13＞peak 18，and their VIP values were greater than 1. Peaks 2，13，15，16 and 18 were positively correlated with antioxidant activity ，and peaks 11 and 12 were negatively correlate d with antioxidant activity ，and the absolute value of standardized regression coefficient were greater than 0.1. CONCLUSIONS ：Fifteen batches of ethanol extracts of M. dodecandrum have antioxidant activity in vitro . The compounds corresponding to common peaks 2，11，12， 13，15，16 and 18 may be the material basis of antioxidant activity of M. dodecandrum .

Identification of a novel TSC2 gene variant in a patient with tuberous sclerosis complex / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a patient diagnosed with tuberous sclerosis complex (TSC).@*METHODS@#Peripheral blood samples of the patient and his parents were collected for the extraction of genomic DNA. Next generation sequencing (NGS) was carried out to detect potential variant, and the result was verified by Sanger sequencing.@*RESULTS@#The patient was found to harbor a heterozygous c.1053delG (p.Glu352SerfsX10) frameshifting variant of the TSC2 gene. The same variant was not found in his unaffected parents and 100 unrelated healthy controls. Based on the American College of Medical Genetics and Genomics guidelines, the variant was predicted to be pathogenic (PVS1+PS2+PM2).@*CONCLUSION@#The novel c.1053delG (p.Glu352SerfsX10) frameshifting variant of the TSC2 gene probably underlay the TSC in this patient.

Histopathological and clinical analysis of 150 cases of dermatofibroma / 中华皮肤科杂志

Objective To analyze histopathological and clinical features of dermatofibroma,and to explore the relationship between them.Methods Clinical and histopathological data were collected from 150 patients with histopathologically confirmed dermatofibroma in Department of Pathology,Shanghai Skin Disease Hospital from September 2017 to August 2018,and analyzed retrospectively.Results Among the 150 patients,65 were males,and 85 were females.Their age was 42 ± 13.8 years,and the course of disease ranged from 3 months to 30 years.Some of the patients had concomitant symptoms,mainly manifesting as itching,some had spontaneous pain and mild tenderness,and 18 patients had a history of injury,insect bite or infection at lesion sites.Skin lesions mainly occurred on the extremities (107 cases,71.3％),and most were solitary (105 cases,70％).Before pathological examinations,102 cases were clinically diagnosed as dermatofibroma,16 as epidermoid cyst,13 as pigmented nevus,3 as keloid,12 as skin mass,1 as malignant melanoma,1 as xanthogranuloma,1 as prurigo nodularis,and 1 as neurofibroma.Among 169 hematoxylin and eosin (HE)-stained sections,25 (14.8％) appeared to be consistent with aneurysmal dermatofibroma,66 (39.1％)with cellular dermatofibroma,36 (21.3％) with sclerosing dermatofibroma,and 22 (13.0％)with epithelioid dermatofibroma.Coexistence of two or more subtypes could be seen in 12 sections.There were also a few new variants,such as dermatofibroma with hyperplastic sweat duct (1 case),deep dermatofibroma (3 cases),dermatofibroma with epithelioid cells intermingled with hyperplastic collagen (1 case).The duration of aneurysmal dermatofibroma varied from 7 months to 30 years,and most manifested as skin masses on the lower extremities.A relatively short course of disease was observed in patients with cellular dermatofibroma,who often visited a hospital several months after the onset,and cellular dermatofibroma was commonly observed on the extremities and frequently accompanied with itching and pain.The duration of sclerosing or atrophic dermatofibroma was usually long for years or decades,and it commonly occurred on the upper limbs without concomitant symptoms.Epithelioid dermatofibroma of varied durations had various clinical manifestations,frequently occurred on the lower limbs without concomitant symptoms.Conclusions The clinical and pathological manifestations of dermatofibroma are diverse.Different dermatofibroma lesions can share similar typical histopathological manifestations,and atypical pathological features can interfere with the diagnosis of dermatofibroma.

Genetic diagnosis of a patient with long-time misdiagnosis of epilepsy / 中华医学遗传学杂志

OBJECTIVE@#To identify pathogenic mutation of TSC1 and TSC2 genes in a patient with long-time misdiagnosis of epilepsy.@*METHODS@#Peripheral blood samples and clinical data of the patient and her 2 parents were collected. Potential mutation of TSC1 and TSC2 genes were detected by direct sequencing.@*RESULTS@#The patient had frequent episodes of epilepsy in addition with Shagreen patches for 10 years. A frame-shifting mutation c.2509_2512delAACA was detected in exon 20 of the TSC1 gene. This same mutation was not found in her unaffected parents.@*CONCLUSION@#The recurrent frame-shifting mutation c.2509_2512delAACA (p.Asn837ValfsX11) of the TSC1 gene probably underlies the disease in this patient.

Genetic diagnosis of a patient with long-time misdiagnosis of epilepsy / 中华医学遗传学杂志

Objective@#To identify pathogenic mutation of TSC1 and TSC2 genes in a patient with long-time misdiagnosis of epilepsy.@*Methods@#Peripheral blood samples and clinical data of the patient and her 2 parents were collected. Potential mutation of TSC1 and TSC2 genes were detected by direct sequencing.@*Results@#The patient had frequent episodes of epilepsy in addition with Shagreen patches for 10 years. A frame-shifting mutation c. 2509_2512delAACA was detected in exon 20 of the TSC1 gene. This same mutation was not found in her unaffected parents.@*Conclusion@#The recurrent frame-shifting mutation c. 2509_2512delAACA (p.Asn837ValfsX11) of the TSC1 gene probably underlies the disease in this patient.

Analysis of TSC gene mutations in five patients with tuberous sclerosis complex / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify pathogenic mutations of TSC1 and TSC2 genes in two familial and one sporadic cases with tuberous sclerosis complex (TSC).</p><p><b>METHODS</b>For five patients and their family members, potential mutations of the TSC1 and TSC2 genes were detected by direct sequencing.</p><p><b>RESULTS</b>For one family, a novel missense mutation c.1964C>T (p.S655F) was detected in the exon 19 of the TSC2 gene. For the sporadic patient, a repeat substitution with deletion mutation c.5238-5255delCATCAAGCGGCTCCGCCA (p.His1746GlnfsX56) was detected in the exon 40 of the TSC2 gene, which led to a stop codon TGA after the 56th amino acids. No mutation was found in another family.</p><p><b>CONCLUSION</b>The missense mutation c.1964C>T(P.S655F) and the substitution with deletion mutation 5238-5255delCATCAAGCGGCTCCGCCA(p.His1746GlnfsX56) of the TSC2 gene probably underlie the disease in the first family and the sporadic case.</p>

Analysis of SLC39A4 gene mutation in a patient with acrodermatitis enteropathica / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect pathogenic mutation of the SLC39A4 gene in a male patient with acrodermatitis enteropathica (AE).</p><p><b>METHODS</b>Peripheral venous blood sample and clinical data from the patient and his parents were collected. One hundred unrelated healthy individuals were recruited as controls. All coding exons and flanking exon-intron sequences of the SLC39A4 gene were analyzed by PCR and direct sequencing.</p><p><b>RESULTS</b>The results revealed that the patient and his mother have both carried a novel frame-shift mutation c.1110InsG (p.Gly370GlyfsX47 to TGA) in exon 6. A novel nonsense mutation c.958C to T (p.Q320X) in exon 5 was also detected in the patient and his father and grandmother. This novel mutation was not detected in the unaffected family members and 100 unrelated healthy controls.</p><p><b>CONCLUSION</b>The novel frame-shift mutation c.1110InsG (p.Gly370GlyfsX47 to TGA) derived from the mother and nonsense mutation c.958C to T (p.Q320X) of the SLC39A4 gene derived from the father may underlie the disease in the patient.</p>

Two novel mutations of the ADAR1 gene associated with dyschromatosis symmetrica hereditaria / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify potential mutation of the ADAR1 gene in a Chinese family and a sporadic case affected with dyschromatosis symmetrica hereditaria(DSH).</p><p><b>METHODS</b>Clinical data and peripheral blood samples from the pedigree and the sporadic patient were collected. Following extraction of genomic DNA, all 15 exons and exon-intron flanking sequences of the ADAR1 gene were amplified by polymerase chain reaction and subjected to direct sequencing.</p><p><b>RESULTS</b>A novel frame-shift mutation c.2638delG (p.Asp880ThrfsX15) from the patients of the pedigree was detected in exon 8 of the ADAR1 gene. And a novel nonsense mutation c.2867C>A (p.Ser956X) was detected in exon 10 of the ADAR1 gene from the sporadic case. Neither mutation was identified among the unaffected family members nor 100 unrelated healthy controls.</p><p><b>CONCLUSION</b>The frame-shift mutation c.2638delG (p.Asp880ThrfsX15) and the nonsense mutation c.2867C>A (p.Ser956X) in the ADAR1 gene probably underlie the DSH in our patients.</p>

Analysis of TSC gene mutation in a patient with tuberous sclerosis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify pathogenic mutation of the TSC1 and TSC2 genes in a patient with tuberous sclerosis.</p><p><b>METHODS</b>Peripheral venous blood samples and clinical data of a pregnant woman with tuberous sclerosis and 4 family members (parents, uncle and husband) were collected. Genomic DNA was extracted. All coding exons of the TSC1 and TSC2 genes and their flanking intronic sequences were amplified by polymerase chain reaction and subjected to direct sequencing.</p><p><b>RESULTS</b>The patient has presented facial angiofibroma and prefrons fibrous plaque for 20 years, and lumbar connective tissue nevus for 10 years. She also had mental retardation but no epilepsy. A novel frame-shift mutation c.4258-4261delTCAG was detected in exon 34 of the TSC2 gene, which had led to a premature stop codon TAG after the 55th amino acids. The same mutation was not found in the unaffected family members and 100 unrelated healthy controls.</p><p><b>CONCLUSION</b>The novel frame-shifting mutation c.4258-4261delTCAG (p.Ser1420GlyfsX55) in the TSC2 gene may be responsible for the disease in the patient.</p>

Brain protection of adenosine preconditioning in infants undergoing radical correction for fallot tetrad / 中国综合临床

Objective To investigate the brain protection of adenosine preconditioning in infants with tetralngy of Fallot (TOF) undergoing open heart surgery. Methods 120 patients with TOF undergoing open heart surgery were randomly divided into two groups: adenosine group and control group. Total dosage of 1.5 mg/kg adenosine was intravenously dripped in the adenosine group before the cardiopulmonary bypass. Serum S-100β and neurone specific enolase (NSE) were measured before the cardipulminary bypass, 30 min and 12 hours after the aorta off clamping. Results At 30 min and 12 hour after opening the aorta, the S-100β[(0. 346±0. 162)μg/L vs. (0.717±0.260) μg/L and (0.541±0.272) μg/L vs. (1.140±0.567) μg/L,P <0.001 ]and NSE[(10.43±5.54) μg/L vs. (15.75±7.96)μg/Land (15.54±7. 98)μg/L vs. (23.67±8.00)μg/L,P<0.001]were both lower in adenosine group than that in control group. Conclusion Adenosine preconditioning can reduce the S-100β and NES in infants with TOF undergoing open heart surgery and has brain protection.