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Objective:To detect mutations in a pedigree containing two brothers with oculocutaneous albinism (OCA) by whole-exome sequencing and Sanger sequencing.Methods:Clinical data were collected from a pedigree with OCA, and DNA was extracted from peripheral blood samples obtained from the proband and other family members. The whole-exome coding region of the proband was directly sequenced by whole-exome sequencing technology to identify potential pathogenic mutations, and Sanger sequencing was conducted to verify the gene mutations.Results:Both the proband and his younger brother presented with generalized white skin, golden-yellow hair, bilateral nystagmus, photophobia, translucent iris, conjunctival congestion, and refractive errors of both eyes. The proband′s parents, grandparents, maternal grandparents, and children were all phenotypically normal, and his parents′ marriage was non-consanguineous. Three heterozygous mutations were identified in the OCA2 gene of both the proband and his younger brother, including a nonsense mutation c.1290T>A, and 2 missense mutations c.1363A>G and c.1204T>C. The mutation c.1204T>C has not been previously reported, and was a novel gene mutation in the OCA2 gene. In addition, 1 heterozygous mutation c.1204T>C was identified in the OCA2 gene in the proband′s father and daughter, 2 heterozygous mutations c.1290T>A and c.1363A>G were found in the OCA2 gene in the proband′s mother, and 1 heterozygous mutation c.1290T>A was identified in the OCA2 gene in the proband′s son and the daughter of the proband′s younger brother.Conclusions:Three gene mutations were identified in the OCA2 gene in the 2 patients with OCA, and the nonsense mutation c.1290T>A may be the pathogenic mutation causing the clinical phenotype of this family. These findings expand the pathogenic mutational spectrum of the OCA gene.
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Objective To explore the protective effects of folic acid on retinas and its anti-oxidative stress mechanism in diabetic mice.Methods Thirty-two 16-week-old SPF degree male db/db mice were randomized into model group and folic acid group,and 16 matched C57BL/KsJ mice were used as controls.Folic acid was used to the mice by oral gavage once per day with the dose of 71 μg/kg (2 ml) for 60 days in the folic acid group,and the same volume of normal saline solution was used in the model group and control group in the same way.The activities,mental state,body weight,and fasting plasma glucose (FPG) of the mice were recorded during experiment.At the end of the intervention,the mice were sacrificed and the retinas and blood sample were obtained.The histopathology of the retinas was examined with hematoxylin-eosin staining;serum homocysteine (Hcy) was detected by ELISA assay;the relative expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected in the retinas by real-time fluorescence quantitative PCR;the relative expressions of B lymphoma 2 protein (bcl-2),bcl-2 related X protein (bax),3-nitrotyrosine (3-NT) and 4-Hydroxynonine (4-HNE) proteins were assayed by Western blot assay;superoxide dismutase (SOD),8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels in the retinas were detected by biochemical kits,and immunofluorescence assay was used to detect the expression of NADPH oxidation 4 (NOX4) in the retinas.The use and care of the experimental animals adhered to the ARVO Statement by the American Association for Vision and Ophthalmology Research and this study protocol was approved by Ethic Committee of Qinghai University (QHDX 2018-35).Results Over the experimental period,The FPG was normal and body weight was gradually increased in the mice of control group.The FPG＞16.7 mmol/L and the mice appeared obese.In the folic acid group,both body mass and FPG of the mice were gradually reduced.At the end of drug administration,serum Hcy concentration of the mice was (27.18± 3.18)μmol/L in the model group,which was significantly higher than (8.28±2.18) μmol/L in the control group and (13.73±2.54) μmol/L in the folic acid group (all at P＜0.05).The retinal structure was intact in the control group,and the retinas were thinning with more capillaries and inflammatory cells in the model group,the thickness of the retinas was increased and the capillaries and inflammatory cells were decreased in the folic acid group.The relative expressions of TNF-α and IL-6 mRNA in the retinas were significantly higher in the model group than those in the control group,and those in the folic acid group were reduced in comparison with the model group (all at P＜0.05).The relative expression of bcl-2 protein in the retinas of folic acid group was lower than that in the control group and higher than that in the model group,and the relative expressions of bax,3-NT and 4-HNE proteins in the retinas of the folic acid group were significantly higher than those in the control group and lower than those in the model group (all at P＜0.05).The T-SOD activity in the folic acid group was significantly stronger than that in the control group and weaker than that in the model group,and the concentrations of 8-OHdG and MDA in the retinas of the folic acid group were significantly reduced in comparison with those of the control group and elevated in comparison with those of the model group (all at P＜0.05).The expressing intensity of NOX4 protein in the retinas of the folic acid group was significantly weaker than that of the model group.Conclusions Folic acid appears aprotective effect on retinal tissue in diabetic mice by reducing serum Hcy,inhibiting oxidative stress and cell apoptosis.
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Objective@#To explore the protective effects of folic acid on retinas and its anti-oxidative stress mechanism in diabetic mice.@*Methods@#Thirty-two 16-week-old SPF degree male db/db mice were randomized into model group and folic acid group, and 16 matched C57BL/KsJ mice were used as controls.Folic acid was used to the mice by oral gavage once per day with the dose of 71 μg/kg (2 ml) for 60 days in the folic acid group, and the same volume of normal saline solution was used in the model group and control group in the same way.The activities, mental state, body weight, and fasting plasma glucose (FPG) of the mice were recorded during experiment.At the end of the intervention, the mice were sacrificed and the retinas and blood sample were obtained.The histopathology of the retinas was examined with hematoxylin- eosin staining; serum homocysteine (Hcy) was detected by ELISA assay; the relative expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected in the retinas by real-time fluorescence quantitative PCR; the relative expressions of B lymphoma 2 protein (bcl-2), bcl-2 related X protein (bax), 3-nitrotyrosine (3-NT) and 4-Hydroxynonine (4-HNE) proteins were assayed by Western blot assay; superoxide dismutase (SOD), 8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels in the retinas were detected by biochemical kits, and immunofluorescence assay was used to detect the expression of NADPH oxidation 4 (NOX4) in the retinas.The use and care of the experimental animals adhered to the ARVO Statement by the American Association for Vision and Ophthalmology Research and this study protocol was approved by Ethic Committee of Qinghai University (QHDX 2018-35).@*Results@#Over the experimental period, The FPG was normal and body weight was gradually increased in the mice of control group.The FPG>16.7 mmol/L and the mice appeared obese.In the folic acid group, both body mass and FPG of the mice were gradually reduced.At the end of drug administration, serum Hcy concentration of the mice was (27.18±3.18)μmol/L in the model group, which was significantly higher than (8.28±2.18)μmol/L in the control group and (13.73±2.54)μmol/L in the folic acid group (all at P<0.05). The retinal structure was intact in the control group, and the retinas were thinning with more capillaries and inflammatory cells in the model group, the thickness of the retinas was increased and the capillaries and inflammatory cells were decreased in the folic acid group.The relative expressions of TNF-α and IL-6 mRNA in the retinas were significantly higher in the model group than those in the control group, and those in the folic acid group were reduced in comparison with the model group (all at P<0.05). The relative expression of bcl-2 protein in the retinas of folic acid group was lower than that in the control group and higher than that in the model group, and the relative expressions of bax, 3-NT and 4-HNE proteins in the retinas of the folic acid group were significantly higher than those in the control group and lower than those in the model group (all at P<0.05 ). The T-SOD activity in the folic acid group was significantly stronger than that in the control group and weaker than that in the model group, and the concentrations of 8-OHdG and MDA in the retinas of the folic acid group were significantly reduced in comparison with those of the control group and elevated in comparison with those of the model group (all at P<0.05). The expressing intensity of NOX4 protein in the retinas of the folic acid group was significantly weaker than that of the model group.@*Conclusions@#Folic acid appears aprotective effect on retinal tissue in diabetic mice by reducing serum Hcy, inhibiting oxidative stress and cell apoptosis.
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Objective To determine the expression of disabled homolog 2 interacting protein (DAB2IP) gene in the basal cell carcinoma (BCC) of the skin,and to investigate its clinical significance.Methods Clinical data were retrospectively analyzed in 105 outpatients and inpatients who received skin mass resection in Department of Dermatology,Guangdong Second Provincial General Hospital and Guangzhou Institute of Dermatology between January 2012 and November 2017.Totally,79 patients with pathologically diagnosed BCC of the skin served as patient group,and 26 patients with pathologically diagnosed skin tag but without other clinical manifestations served as control group.Immunohistochemical staining was performed to determine the expression of DAB2IP in the two groups,and correlations of the DAB2IP expression with the clinical phenotype and pathological features of BCC of the skin were analyzed.Statistical analysis was carried out with SPSS21.0 software by using chi-square test for the comparison of enumeration data.Results The protein expression of DAB2IP was observed in 11 (42.3％) of 26 patients in the control group,as well as in 74 (93.7％) of 79 patients in the patient group,and there was a significant difference in the positive rate of DAB2IP protein between the two groups (x2 =33.50,P ＜ 0.05).The expression of DAB2IP was uncorrelated with gender or age of patients with BCC of the skin,or with the tumor size (all P ＞ 0.05).The positive rate of DAB2IP protein significantly differed between the patients with superficial BCC (5/7) and those with invasive BCC (95.8％,69/72;x2 =6.47,P ＜ 0.05).Of the 79 patients with BCC of the skin,Ki-67 protein was detected in 31 (39.2％),and the cancer cells expressing Ki-67 protein also expressed DAB2IP protein.Conclusion The expression of DAB2IP increases in BCC of the skin,which may be associated with the occurrence and infiltration of BCC of the skin.
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Objective To study the ultramicrostructure change and keartin(KRT1) expression in skin lesion of symmetrical acral keratoderma(SAK) .Methods Thirteen cases of SAK in the First Affiliated Hospital of Fujian Medical University and the outpatient department of the Dongguan Municipal Sixth People′s Hospital were selected as the study subjects .The histopathological samples were taken from the wrist site .The retinoic acid preparation or corticosteroid preparation or Chinese medicine preparation were not externally used within 2 months before taking skin lesion sample .The healthy control skin samples were the normal skin in 12 cases by plastic surgical resection .The ultramicrostructural change were observed by the transmission electron microscopy .The KRT1 expression in skin lesion of 13 cases of SAK and healthy skin tissue of 12 cases were measured by immunohistochemistry method .Results The SAK ultramicrostructures manifested by the interruption of keratinizing envelope continuity in horny layer , and remarkable aggregation of keratin filament in upper stratum spinosum and surrounding nucleus of granular layer .KRT1 was ex-pressed in the cells of SAK skin lesion and basal layer ,spinous layer ,granular layer and horny layer .The cytoplasm and cytomem-brane staining was common .The KRT1 expression in skin lesion was significantly higher than that in normal skin (t=2 .210 ,P=0 .038) .Conclusion The ultramicrostructure features of SAK skin lesion are abnormal differentiation of epidermis keratin fila-ments ,which might be related with overexpression of KRT 1 .
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Objective To detect mutations in the filaggrin (FLG) gene and expressions of FLG and loricrin in patients with ichthyosis vulgaris (Ⅳ),and to investigate their clinical significance.Methods Tissue specimens were resected from the skin lesions of 10 patients with Ⅳ and normal skin of 14 healthy human controls,and immunohistochemical SP method was used to detect the expressions of filaggrin and loricrin.The expression intensity was determined by the Image-Pro Plus (IPP) software,and expressed as positive units (PU).Blood samples were collected from 10 patients of Han nationality with Ⅳ and 100 healthy human controls followed by DNA extraction.PCR and DNA sequencing were performed to detect the presence of 13 mutations (3321delA,441delA,1249insG,E1795X,S3296X,R501X,2282de14,R2447X,S2889X,7945delA,3702delG,Q2417X,R4307X) in the FLG gene.Results FLG was mainly expressed in the cytoplasm of keratinocytes in the stratum corneum,granular layer,prickle layer and basal layer,and loricrin was observed in the cytoplasm and nuclei of keratinocytes in the granular layer,prickle layer and basal layer,in both the lesional and normal skin.Compared with the normal skin,the lesional skin showed significantly weaker expressions of FLG (0.208 2 ± 0.008 0 vs.0.230 0 ± 0.0228,t =3.30,P ＜ 0.01) and loricrin (0.137 0 ± 0.011 2 vs.0.149 3 ± 0.007 3,t =3.07,P ＜ 0.01).Sequencing analysis identified two mutations,including 3321delA in 7 patients and 441delA in 2 patients.No mutations were detected in the healthy controls.Conclusions The 3321delA and 441delA mutations in the FLG gene may represent the most frequent genetic cause of Ⅳ in patients of Han nationality.The low expressions of FLG and loricrin may be associated with the impairment of skin barrier function in patients with Ⅳ.
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Objective To analyze the clinical,demographic and pathologic features of symmetric acral keratoderma.Methods Totally,62 outpatients with symmetric acral keratoderma were collected at the Dongguan Hospital for Chronic Disease Prevention and Control from May 2003 to June 2012.Data on these patients,including demographic information,family history,physical and pathological examination findings,were reviewed.Results There were 55 males and 7 females among these patients.The age at onset and disease duration varied from 4 to 53 years (average: 24.02 years) and from 15 days to 10 years (average: 26.65 years) respectively.Characterized manifestation was brown keratinized patches on the dorsum of hands and digits,wrists,elbows,knees,ankles,and along the lateral margin of palms,which became white 3 to 6 minutes after immersing in water.Lesions usually resolved spontaneously in winter.Microscopic examination of lesions for fungal elements was negative.Histopathology revealed epidermal hyperkeratosis,acanthosis and mild papillomatous hyperplasia as well as dermal infiltrates with a few lymphocytes.Conclusion Symmetric acral keratoderma is a skin disease characterized by symmetric acral keratosis with apparent seasonality.
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A 10-year-old boy presented with a 3-year history of erythematous flat keratotic papules and brown-yellow, nail-like prominent keratotic plaques all over the body surface. Dermatological examination showed verrucous or nail-like prominence over multiple erythematous keratotic plaques on the head, face,trunk and limbs. The lesions, most of which confluenced, were covered with brown-yellow and greasy crusts,and gave a porcupine-like appearance. Skin biopsy of lesions from the back revealed epidermal hyperkeratosis,focal columnar parakeratosis, acanthosis, few acantholytic and dyskeratotic cells in stratum corneum, irregular upward proliferation of dermal papilla cells, and a superficial perivascular lymphocytic infiltration. A diagnosis of ichthyosis hystrix was established based on the histopathological findings. The boy was treated with oral acitretin and topical 0.1% acitretin cream for 8 years. The initial and maximum dose of oral acitretin was 0.5 mg·kg-1·d-1 and 1 mg·kg-1·d-1, respectively. Liver and kidney function, body height and weight were examined during the treatment, and no side effect was observed except for skin xerosis.
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Objective To evaluate the therapeutic effects and safety of acitretin for severe inherited keratodermas in children and adolescents. Methods Acitretin was given to 23 children and adolescents with either lamellar ichthyosis, bulbous ichthyosiform erythroderma, pityriasis rubra pillars, progressive sym- metrical erythrokeratoderma, keratitis ichthyosis deafness syndrome, generalized porokeratosis, inflammatory liner verrucous epidermal nevus, ichthyosis hystrix and non-bullous ichthyosiform erythroderma. The thera- peutic dosage was 0.67-1.07 mg/(kg?d),and maintenance dosage 0.08-0.94 mg/(kg?d).The effects on the patients' growth and development of the drug were evaluated based on the changes of body weight and height in the children. The total follow-up period was 6-35 months in an interval of 1-3 months. Results The considerable overall improvement was achieved after 1-6 months' treatment, with an overall clinical cure rate of 82.6%. Only one case responded poorly to the therapy. The excellent responses were observed in patients with bulbous ichthyosiform erythroderma, lamellar ichthyosis, and pityriasis rubra pillars, etc, and the much poor responses in ichthyosis hystrix. The most frequent adverse reaction was mild to moderate dry lips (65.2%),the next were pruritus(39.1%),skin fragility(34.8%),and dry mouth(30.4%).The less frequent adverse reactions were alopecia(13%),anorexia(8.7%),headache (4.3%) and hypoacusis (4.3%).No effects on the growth and development were found in those children during the follow up period. Conclusions The considerable overall improvement is achieved with the acitretin therapy for children and adolescents with inherited keratodermas, with only mild to moderate adverse reactions and no effects on the growth and development in the children.
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Objective To investigate the clinical effect and safety of Chinese medicinal liniment pourmask and smearing in the treatment of acne. Methods Two-hundred and nine cases of acne were divided into two groups: treatment group (106 cases) was treated with Chinese medicinal liniment pourmask and smearing for 1 to 4 weeks, and control group was treated with gypsum fibrosum pourmask combining chloramphenicol and salicyic acid tincture for 1 to 4 weeks. Results At 8th week the cure rate, effective rate and improved rate in the treatment group and control group were 70.75 %, 29.25 %, 0 % and 34.95 %, 42.72 %, and 22.33 % respectively. Side effects were not found in the treatment group. Ten cases (9.71 %) in control group had contact dermatitis. Conclusion Chinese medicinal liniment pourmask and smearing have a good efficacy and safety in the treatment of acne.
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Objective To detect the mutations of GJB2 and GJB6 genes in the first Chinese case of keratitis, ichthyosis and deafness (KID) syndrome. Methods Genomic DNA was extracted from the patient with KID syndrome and his family members. All encoding exons and adjacent splice sites of the GJB2 and GJB6 genes were amplified by PCR. Mutation scanning was carried out by direct bidirectional DNA sequencing. Results No mutation was found in GJB6. A G148A mutation was found at exon2 of GJB2 in the patient, which caused a change from aspartic acid to asparagine at codon 50(D50N). Conclusion This case of KID syndrome may be caused by the mutation in GJB2.