Objective To analyze KIT gene mutations in one patient with diffuse cutaneous mastocytosis (DCM),and to provide a basis for the prediction of prognosis and selection of treatment.Methods Clinical data were collected from a boy with DCM.Peripheral blood samples were obtained from the patient,his parents and 200 unrelated healthy human controls.PCR was performed to amplify 21 coding exons and their flanking sequences of the KIT gene followed by DNA sequencing.Results A heterozygous missense mutation (c.1526A ＞ T),which leads to the mutation p.Lys509Ile,was detected in the KIT gene of the patient,but not in his parents or the healthy controls.Conclusion The heterozygous missense mutation p.Lys509Ile in the KIT gene may be a cause of DCM.

Objective To report two cases of Nagashima-type palmoplantar keratoderma(NPPK), and to identify mutations in the SERPINB7 gene. Methods Clinical data were collected from two patients with NPPK and their parents, and peripheral blood samples were obtained from the two patients, their parents and 200 unrelated healthy controls. Genomic DNA was extracted from these blood samples. PCR was performed to amplify 8 exons and their flanking sequences of the SERPINB7 gene followed by DNA sequencing. Results A homozygous mutation (c.796C > T), which led to the formation of a premature termination codon at amino acid position 266 (p.R266*), was identified in both of the two patients. However, the patients′ healthy parents were heterozygous carriers of the mutation(c.796C > T). No mutation was found in the unrelated healthy controls. Conclusion The mutation c.796C > T in the SERPINB7 gene may be responsible for NPPK in the two patients.

Objective To assess mutations in the ALDH3A2 gene in two patients with Sj(o)gren-Larsson syndrome manifesting primarily as congenital ichthyosis,mental retardation and spastic paraplegia.Methods Two patients,a 2-year-old girl and a 1.5-year-old boy,with Sj(o)gren-Larsson syndrome were included in this study.None of their family members suffered from this disease.Peripheral blood samples were collected from the two patients,their family members (an elder brother and both parents),and 100 unrelated healthy controls.DNA was extracted from the blood samples,and subjected to PCR for the amplification of 10 encoding exons and their flanking sequences of the ALDH3A2 gene followed by DNA sequencing.Results A homozygous missense mutation c.325G ＞ A,which leads to the substitution of glycine by arginine at position 109,was detected in the ALDH3A2 gene of patient 1,whose parents and elder brother were heterozygous carriers of this mutation.The patient 2 carried compound heterozygous mutations,including c.1157A ＞ G (p.Asn386Ser) inherited from his father and c.1294A ＞ T (p.Arg432X) inherited from his mother.None of these mutations was detected in the unrelated healthy controls.Conclusion The homozygous mutation p.Gly109Arg and compound heterozygous mutations p.Asn386Ser and p.Arg432X present in these patients may be associated with clinical phenotypes of Sj(o)grenLarsson syndrome.

Objective To detect the mutations of COL7A1 gene in three cases of dystrophic epidermolysis bullosa pruriginosa (DEBP). Methods Clinical data were collected from 3 patients with DEBP. Skin lesions were obtained from these patients and subjected to transmission electron microscopy. DNA was extracted from the peripheral blood of the 3 patients, their 16 relatives, and 150 unrelated normal human controls, and PCR was performed to amplify all the exons and flanking sequences of COL7A1 gene followed by sequencing.Results The patient 1 and 2 had family history, whereas the case 3 was sporadic. Transmission electron microscopy showed tissue cleavage beneath lamina densa in case 1 and slightly decreased anchoring fibrils in some areas of the lesions in case 1 and 3. Three heterozygous mutations of COL7A1 gene, i.e., c. G6734T, c.G6859A and c. G5318T, which leaded to three amino acid mutations, i.e., p. G2245V, p. G1773V and p. G2287R, were found in patient 1, 2 and 3 respectively. Of them, p. G2245V and p. G1773V were novel mutations. The mutations strictly cosegregated with the phenotype in the patients of family 1 and 2. No mutation was detected in the unaffected parents of patient 3 or the 150 unrelated healthy controls. Conclusions The p. G2245V, p. G2287Rand p. G1773V mutations of COL7A1 gene may be responsible for the phenotype of DEBP in the three cases,and of them, p. G2245V and p. G1773V have never been reported.

Objective To detect the mutation of CARD15 gene in a patient with sarcoidosis and tuberculosis.Methods Clinical data were collected from a 32-year-old male patient with early-onset sarcoidosis and tuberculosis.Peripheral blood was obtained from the patient,both of his parents,and 102 healthy controls.All the 12 exons of the coding regions as well as flanking intronic sequences of the CARD15 gene were amplified by PCR followed by direct sequencing.The resulted sequences were blasted against the reference sequences of CARD15 gene.Results Both clinical features and histopathological findings of the patient were consistent with sarcoidosis.Furthermore,the patient presented with flexion contractures of fingers and toes,as well as iridocyclitis.A heterozygous missense recurrent mutation c.1000C ＞ T (p.R334W) was detected in exon 4 of the CARD15 gene in the patient,but not in either of his parents or any of the 102 healthy controls.Conclusions A p.R334W mutation in the CARD15 is identified in the patient,which may be responsible for the clinical phenotype of earlyonset sarcoidosis.Gene analysis may be a useful method to clarify the etiology of early-onset sarcoidosis.

Objective To study cutaneous ultrastructural changes and FERMT1 gene mutations in a patient with Kindler syndrome. Methods Clinical data were collected, and tissue samples obtained from the lesions of poikiloderma were observed by using transmission electron microscopy. Fifteen coding exons and their flanking sequences of the FERMT1 gene were amplified by PCR and DNA sequencing was followed.Results Reduplication of lamina densa was seen between the dermal-epidermal junctions of the lesional skin. The patient was found to be homozygous for a novel splice-site mutation (IVS9 + 1G ＞ A) in FERMT1 gene, and his parents were heterozygous for it. The mutation was undetected in fifty normal control individuals.Conclusions Transmission electron microscopy may serve as an ancillary examination for the diagnosis of Kindler syndrome. The IVS9+1G＞A mutation of FERMT1 gene may contribute to the clinical phenotype of Kindler syndrome in this patient.

Objective To observe the ultrastructural features of recessive dystrophic epidermolysis bullosa inversa(RDEB-Ⅰ)and to detect the mutations of COL7A1 gene in a family with RDEB-Ⅰ.Methods A 24-year-old male patient complained of recurrent vesicles in the skin for 24 years.The lesions began as generalized pruritic vesicles and bullae soon after birth,with a predilection for areas subject to friction,and showed a trend to be worse in summer but mild in winter.No photosensitivity was observed.When he was 3 to 4 years old,the lesions were decreased in number,with the only involvement of the trunk and abdomen;thereafter,the lesions were improved year by year.The patient suffered from nephritis at the age of 5 years,which progressed into renal failure at the age of 15 years.He received renal transplantation and was given long-term oral tacrolimus and mycophenolate mofetil,which leaded to an improvement in the lesions.The family history was unremarkable,and the marriage between her parents was not consanguineous.Dermatological examination revealed large areas of irregularly-marginated,hypopigmented,atrophic scar on the waist,back and abdomen with onychodystrophy involving multiple nails.No vesicles were observed.Immunofluorescence antigen mapping and transmission electron microscopy were conducted to observe the expression of type Ⅶ collagen in and ultrastructure of cutaneous lesions from the patient.Venous blood samples were obtained from the patient as well as his parents and 3 sisters,and drill biopsy specimens were obtained from the margin of vesicular lesions and unaffected anterior tibial skin of the patient.DNA specimens were obtained from the blood samples of the family members and 150 unrelated healthy controls,and RNA was extracted from the biopsy samples of the patient.PCR and direct sequencing were carried out to detect mutations in COL7A1 gene,and reverse transcription-PCR was conducted to confirm the mutation at mRNA level.Results Skin cleavage was observed under lamina densa in the dermis,with a decrease in anchoring fibrils and expression of type Ⅶ collagen in the lesions of the patient.A heterozygous synonymous mutation c.C5499T which created a new splicing site and leaded to a premature terminal codon,as well as a heterozygous missense mutation c.C6205T(C-T transition at codon 2069:CGT to TGT)which leaded to the substitution of arginine by cysteine,were identified in the COL7A1 gene of the proband and all of his sisters,but not in any of the unrelated controls.The c.C5499T and c.C6205T mutations were inherited from the patient's father and mother respectively.Conclusion The compound heterozygous mutations c.C6205T and c.C5499T may be responsible for RDEB-Ⅰ in this patient.

Objective To detect mutations of the ABCA12 gene in 2 Chinese families with autosomal recessive congenital ichthyosis (ARCI).Methods According to the typical clinical manifestations,two probands were diagnosed with ARCI.DNA was extracted from the peripheral blood samples collected from the patients and their parents.High-throughput sequencing was conducted by using multi-gene array for genetic skin disorders to determine mutation sites in the probands,and then DNA isolated from the probands and their parents were bidirectionally verified by Sanger sequencing.Results Two compound heterozygous mutations (c.2759A＞G and c.7004A＞G) in the ABCA12 gene were found in the proband 1,and another two compound heterozygous mutations (c.6163_6164insT and c.7406G＞A) were identified in the proband 2.The parents of the two probands were heterozygous carriers of one of the two mutations in the ABCA12 gene.Function prediction for the 4 mutations showed that all of the 3 missense mutations (c.2759A＞G,c.7004A＞G and c.7406G＞A) may exert pathogenic effect,and fragnin encoded by the frameshift mutation c.6163_6164insT may also affect protein function,c.2759A＞G and c.6163_6164insT were newly identified mutation sites.Conclusion The compound heterozygous mutations in the ABCA 12 gene are the causative mutations responsible for ARCI in the two probands of the two pedigrees.

Objective:To understand the genome characteristics of group A rotavirus (RVA) strains among hospitalized children under 5 years of age with acute diarrhea in Fuzhou sentinel hospital in 2020.Methods:The ELISA method was used for screening RVA-positive stool samples of hospitalized children under 5 years of age, then 11 gene segments of RVA-positive samples were sequenced and typed after amplification by RT-PCR, and their homology and phylogenetic analysis were performed by Molecular Evolutionary Genetics Analysis (MEGA).Results:Twenty RVA whole genome sequences were successfully obtained, including 4 kinds of G/P gene combinations-G9P[8] (55%), G3P[8] (25%), G8P[8] (15%) and G2P[4] (5%). DS-1-like reassortant strains accounted for 40% of the whole genomes. Strains of the same type have high sequence homology, are closely related to the virus strains that currently circulating in the world. There are mutations at multiple important antigenic sites on VP7, VP4 and NSP4 fragments. The variation of amino acid substitutions of VP7, VP4 and NSP4 fragments is complicated, and there are many amino acid substitutions in the antigenic regions.Conclusions:G3P[8] and G8P[8] DS-1-like reassortants were detected for the first time in Fuzhou, amino acid substitutions were observed in the antigenic regions of the VP7, VP4 and NSP4 gene. Due to the emergence of uncommon DS-1-like reassortant strains and multiple important antigenic regions substitutions, it is necessary to continuously monitoring genome characteristics of RVA strains to provide scientific evidence for the pandemic prevention and vaccine immunization strategies.

Objective To explore the efficacy and safety of ultrasound-guided radiofrequency ablation combined with foam sclerotherapy in the treatment of primary small saphenous vein varicosities.Methods A total of 46 patients with primary small saphenous vein varicosities in our department from November 2021 to November 2022 were retrospectively analyzed.Depending on the patient's choice of surgical method,they underwent radiofrequency ablation combined with sclerotherapy(observation group,n=23)or high ligation and stripping of the saphenous vein(control group,n=23).The surgical time,intraoperative blood loss,length of hospital stay,postoperative Venous Clinical Severity Score(VCSS),incidence of postoperative complications,treatment satisfaction,and recurrence were compared between the two groups.Results Compared with the control group,the observation group had shorter surgical time[(56.1±6.0)min vs.(81.7±10.6)min,t=-10.128,P=0.000],less intraoperative blood loss[(27.0±4.1)ml vs.(41.4±4.8)ml,t=-11.016,P=0.000],shorter hospitalization time[(2.0±0.7)dvs.(5.6±0.9)d,t=-14.319,P=0.000],lower VCSS scores at 3,6,and 12 months after surgery[(7.4±2.0)points vs.(8.9±2.5)points,t=-2.165,P=0.036;(5.3±1.8)points vs.(6.5±2.2)points,t=-2.149,P=0.037;(2.6±1.3)points vs.(4.0±1.8)points,t=-2.912,P=0.006],higher satisfaction[satisfied,somewhat satisfied,dissatisfied in 18,4,and 1 cases in the observation group and 8,10,and 5 cases in the control group,respectively,Z=-2.967,P=0.003].There were no statistically significant differences in the complication incidence and recurrence rate between the two groups(P＞0.05).Conclusion Ultrasound-guided radiofrequency ablation combined with foam sclerotherapy in the treatment of primary small saphenous vein varicosities is safe and effective.