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Objective:To construct "Expert consensus on diagnosis and treatment of chronic inducible urticaria in China (2023) " based on the Delphi method, and to provide a methodological basis for consensus construction.Methods:After systematic search and evaluation of the literature related to chronic inducible urticaria, the first draft of "Expert consensus on diagnosis and treatment of chronic inducible urticaria in China (2023) " was written, and a questionnaire was designed for expert consultation. A representative sample of 25 experts was selected to conduct two rounds of correspondence consultation via electronic questionnaire in strict accordance with the Delphi method, and the content of the consensus was revised and improved according to the consultation results.Results:The response rates in the two rounds of questionnaire consultation were both 100%, and the expert authority coefficient was 0.92 ± 0.09. In the first round of consultation, the coefficients of variation (CV values) of 9 items were greater than 20%, and the mean agreement degree of 3 items was less than 7 points; in the second round of consultation, the CV values of all items were less than 15%, the agreement degree of the above 3 items whose mean agreement degree was less than 7 points in the first round of consultation all rose to over 7 points, and the median agreement degree of all items was greater than or equal to 8 points. Reliability analysis of the two rounds of questionnaire results showed that the Cronbach α coefficient and standardized Cronbach α coefficient were both greater than 0.9; the P values in the agreement tests by using Kendall′s coefficient of concordance for the two rounds of questionnaire results were both less than 0.001, and the Kendall′s coefficients of concordance were 0.170 and 0.219 in the first and second rounds of questionnaire consultation, respectively. Conclusion:The Delphi method-based "Expert consensus on diagnosis and treatment of chronic inducible urticaria in China (2023) " is highly representative, authoritative and reliable; this study also provides a methodological reference for the formulation and research of consensus.
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Objective:To compare efficacy and safety of 308-nm SQ light-emitting diode (LED) light versus 308-nm excimer light in the treatment of facial vitiligo.Methods:Patients with stable facial vitiligo were retrospectively collected from Department of Physical Therapy, Hospital of Dermatology, Chinese Academy of Medical Sciences from June 2018 to June 2020, who received treatment with 308-nm SQ LED light (LED group) or 308-nm excimer light (excimer light group). The treatment was performed once or twice a week, and patients who had received more than 8 sessions of treatment were included in the analysis of efficacy and safety. Statistical analysis was carried out by using chi-square test.Results:Totally, 68 patients with 90 lesions were enrolled into the LED group, including 36 males and 32 females, aged 25.01 ± 13.37 years; 20 patients with 28 lesions were enrolled into the excimer light group, including 13 males and 7 females, aged 27.15 ± 14.30 years. After 8 and 16 sessions of treatment, there was no significant difference in the response rate between the LED group (23.33%, 46.67%, respectively) and excimer light group (14.29%, 46.43%, χ2 = 1.05, < 0.001, respectively, both P > 0.05). During the treatment, 36 (52.94%) patients in the LED group developed persistent erythema, 17 (85%) in the excimer light group developed persistent erythema or blisters. The incidence of adverse reactions was significantly lower in the LED group than in the excimer light group ( χ2 = 16.43, P < 0.001) . Conclusion:Compared with the 308-nm excimer light, the 308-nm SQ LED light showed similar effect but higher safety for the treatment of facial vitiligo.
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Dermatitis and eczematous disorders are very common in dermatological clinic. Due to variations in understanding of these diseases, there are some confusions and controversies about the diagnostic terms of "dermatitis" and "eczema" in China and other countries. With the progress in research on dermatitis and eczematous disorders, the understanding of some diseases with unclear causes or etiologies has gradually been improved, and it has been recognized that neither "dermatitis" nor "eczema" should be regarded as an independent clinical diagnostic term. Standardizing the diagnostic process and terminology for related diseases can lay a foundation for better classification, management, epidemiological research, and targeted treatment and prevention of these diseases.
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Beta (β)-thalassemia is one of the most common hemoglobinopathies worldwide, creating major public health problems and social burdens in many regions. Screening for β-thalassemia carriers is crucial for controlling this condition. To investigate the effectiveness of mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) for screening β-thalassemia, retrospective data were analyzed for 6,779 β-thalassemia carriers subjected to genetic testing following thalassemia screening in Guangdong province between January 2018 and December 2019. Prevalent mutations observed included CD41/42 (-TTCT) (38.43%), IVS-II-654 (C > T) (25.71%), -28 (A > G) (15.78%), CD17 (AAG > TAG) (10.03%), and β
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Adult , Female , Humans , Male , Middle Aged , Young Adult , China , Erythrocyte Indices , Mass Screening , Mutation , beta-Thalassemia/geneticsABSTRACT
Objective:To evaluate the effect of resveratrol on apoptosis and cell cycle of human epidermal keratinocytes (HEKs) after ultraviolet B (UVB) irradiation.Methods:After 12-hour treatment with 0 (control group) , 25, 50, 100, 150 and 200 μmol/L resveratrol, cell counting kit-8 (CCK8) assay, bromodeoxyuridine (BrdU) assay and lactate dehydrogenase (LDH) assay were performed to evaluate the effect of resveratrol on proliferative activity and death of HEKs. Some HEKs were divided into 4 groups: normal control group receiving no treatment, resveratrol group treated with 100 μmol/L resveratrol, UVB group irradiated with 50 mJ/cm 2 UVB, and UVB+resveratrol group irradiated with 50 mJ/cm 2 UVB followed by 12-hour treatment with 100 μmol/L resveratrol. Western blot analysis was conducted to determine the expression of apoptosis-related proteins poly (ADP-ribose) polymerase (PARP) and caspase-3, as well as cell cycle-related proteins cyclin B1 and cyclin E1, and flow cytometry to detect the apoptosis and determine cell cycle distribution. One-way analysis of variance was used for comparisons among multiple groups, and least significant difference- t test for multiple comparisons. Results:The proliferative activity of HEKs significantly differed among the 25-, 50-, 100-, 150-, 200-μmol/L resveratrol groups and control group ( F=46.52, P < 0.001) , and was significantly lower in these resveratrol groups than in the control group (all P < 0.001) ; the DNA synthesis activity of HEKs was significantly lower in the 100-μmol/L resveratrol group (0.761 ± 0.141) than in the control group (2.226 ± 0.141, t=17.92, P < 0.001) ; there was no significant difference in cell death rate among the 25-, 50-, 100- and 200-μmol/L resveratrol groups ( F=1.938, P > 0.05) . After treatment with or without UVB and/or resveratrol, there were significant differences in the apoptosis rate, proportion of cells at G1, G2 and S phases, expression of PARP, caspase-3, cyclin B1 and cyclin E1 among the 4 groups (all P < 0.05) . The UVB group showed significantly increased apoptosis rates (24.69% ± 3.54%) and cleavage levels of PARP and caspase-3 (2.190 ± 0.349, 0.090 ± 0.016, respectively) compared with the normal control group and UVB+resveratrol group (4.00% ± 0.81%, 0.926 ± 0.040, 0.024 ± 0.019, respectively, all P < 0.05) ; the UVB group showed significantly decreased protein expression of cyclin E1 and cyclin B1 (0.116 ± 0.017, 0.775 ± 0.080, respectively) compared with the normal control group, and the UVB+resveratrol group showed significantly increased expression of cyclin E1 (0.287 ± 0.047) , but decreased expression of cyclin B1 (0.504 ± 0.009) compared with the UVB group (all P < 0.05) ; the UVB group showed significantly decreased proportion of S-phase cells, but increased proportion of G1- and G2-phase cells compared with the UVB+resveratrol group (all P < 0.05) . Conclusion:Resveratrol can decrease the proliferative activity of HEKs, inhibit apoptosis induced by UVB radiation, and regulate cell cycle progression.
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Thalassemia is a group of genetically heterogeneous diseases characterized by hemolytic anemia. To investigate molecular characteristics of α- and β-thalassemia among young individuals of marriageable age in Guangdong Province, 24,788 subjects with suspected thalassemia were genetically tested for α- and β-thalassemia by Gap-PCR and reverse dot blot during 2018-2019. For suspected rare thalassemia cases, DNA sequencing was performed to identify rare and unknown thalassemia gene mutations. A total of 14,346 thalassemia carriers were detected, including 7,556 cases of α-thalassemia with 25 genotypes and 8 α-gene mutations identified, 5,860 cases of β-thalassemia with 18 genotypes and 18 β-gene mutations identified, and 930 cases of compound α/β-thalassemia. Among them, the frequency of --
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Adult , Female , Humans , Male , Middle Aged , Young Adult , China , Genotype , Mutation , Sequence Analysis, DNA , alpha-Thalassemia/genetics , beta-Thalassemia/geneticsABSTRACT
Objective:To evaluate the efficacy of acrivastine alone or in combination with loratadine in the treatment of chronic refractory urticaria.Methods:From March 2017 to December 2018, a multicenter, randomized, controlled clinical study was conducted in 4 centers. Patients with chronic refractory urticaria were randomly divided into two groups, i.e., combined treatment group receiving oral acrivastine capsules 8 mg thrice a day plus oral loratadine tablets 10 mg once a day, and acrivastine alone group receiving oral acrivastine capsules 8 mg thrice a day plus a placebo 10 mg once a day. The course of treatment was 4 weeks. Visits were scheduled at baseline and after 1, 2 and 4 weeks of treatment. At the same time, clinical data were collected, and adverse events were recorded. Symptom scores were evaluated based on degree of itching, number and size of wheals, duration of each attack and number of attacks per week, and symptom score reduce index (SSRI) was used to evaluate the efficacy. Repeated measures analysis of variance and chi-square test were used to evaluate the efficacy and safety.Results:Fifty-three patients in the combined treatment group and 59 in the acrivastine alone group were included in the efficacy analysis. Before treatment, there was no significant difference in symptom score or visual analogue score between the two groups. After 2 weeks of treatment, 19 patients were cured and 10 achieved marked improvement in the combined treatment group, with a response rate of 54.72%; 15 were cured and 6 achieved marked improvement in the acrivastine alone group, with a response rate of 35.59%. After 4 weeks of treatment, 23 patients were cured and 9 achieved marked improvement in the combined treatment group, with a response rate of 60.38%; 20 were cured and 2 achieved marked improvement in the acrivastine alone group, with a response rate of 37.29%. After 2 and 4 weeks of treatment, the response rates were significantly higher in the combined treatment group than in the acrivastine alone group ( χ2 = 4.13, 5.96 respectively, both P < 0.05) . The SSRI significantly differed among different follow-up time points, as well as between the 2 groups ( F = 8.62, 4.38 respectively, both P < 0.05) . Multivariate analysis of variance showed that SSRI was significantly higher in the combined treatment group (0.63 ± 0.05, 0.68 ± 0.05, respectively) than in the acrivastine alone group (0.47 ± 0.05, 0.51 ± 0.05, respectively) after 2 and 4 weeks of treatment (both P < 0.05) . Drug-related adverse reactions, including drowsiness, stomach upsets, headache and liver function abnormality, occurred in 7 patients in the combined treatment group, as well as in 3 in the acrivastine alone group. Conclusion:Acrivastine is safe and effective for the treatment of chronic refractory urticaria, and acrivastine combined with loratadine can markedly improve the efficacy.
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The COVID-19 outbreak may have some impact on the use of biologics in psoriatic patients because immunosuppressive effects of biologics may potentially alter the susceptibility of patients to the virus, deteriorate the condition of infected patients or even change the prognosis of infection. According to currently available recommendations from international psoriasis academic organizations and specialists, as well as specific situation in China, the authors provide some guidance on the use of biologics for psoriatic patients undergoing or planning to undergo treatment with biologics, those with low or high risk of infection, and for those with or without COVID-19 infection, so as to provide references for clinical practice.
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Objective To investigate the role of proprotein convertase subtilisin/kexin type 9 (PCSK9) in the pathogenesis of psoriasis by detecting the level of PCSK9 in the plasma of patients with psoriasis and evaluating its effect on the secretion of interferon gamma (IFN-γ) and interleukin-17A (IL-17A) by peripheral CD4+ T cells.Methods Totally,30 outpatients with psoriasis vulgaris and 30 healthy volunteers (controls) were enrolled from Hospital for Skin Diseases,Chinese Academy of Medical Sciences between February 2016 and December 2017.Of the 30 patients,16 were males,and 14 were females.Their age varied from 18 to 66 years,and the course of disease ranged from 1 month to 15 years.Peripheral venous blood samples were obtained from the patients and controls,and the plasma and was performed to measure mRNA expression of PCSK9 in the PBMC,and enzyme-linked immunosorbent assay (ELISA) to determine the concentration of PCSK9 in the plasma.Peripheral CD4+ T cells were isolated from the PBMC by magnetic bead method,and divided into 2 groups to be co-cultured with (experiment group) or without PCSK9 protein (control group).After 24-hour treatment,ELISA was conducted to detect the levels of IFN-γ and IL-17A in the culture supernatant.Statistical analysis was carried out by using two-sample t test for the comparison between the two groups,and Pearson correlation analysis for analyzing correlations between the plasma level of PCSK9 and psoriasis area and severity index (PASI) score in the patients with psoriasis.Results PCSK9 mRNA expression was undetected in the PBMC of the patients with psoriasis and controls.The plasma level of PCSK9 was significantly higher in the patients with psoriasis ([243.58 ± 11.91] μg/L) than in the healthy controls ([199.74 ± 31.09] μg/L,t =5.761,P ＜ 0.001).After co-culture of the peripheral CD4+ T cells from patients with PCSK9 protein,the levels of IFN-γ and IL-17A both significantly increased ([6 150.00 ± 212.13] ng/L,[1 532.00 ± 11.31] ng/L,respectively) compared with the control group co-cultured without PCSK9 protein ([4 650.00 ± 212.13] ng/L,[698.5 ± 266.58] ng/L,respectively;t =7.071,4.418 respectively,both P ＜ 0.05).IFN-γand IL-17A were undetected in the culture supernatant of CD4+ T cells from the healthy controls in the experiment group or control group.Conclusion The plasma level of PCSK9 increases in patients with psoriasis,which may be involved in the pathogenesis of psoriasis by activating peripheral CD4+ T cells.
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Psoriasis is a common chronic inflammatory skin disease,and commonly considered as a T cell-mediated abnormal immune disease with genetic predisposition at present.Recent studies have indicated that microRNAs and lncRNAs play important roles in occurrence and development of a variety of inflammatory immune diseases.It has been confirmed that microRNAs and lncRNAs participate in the occurrence of psoriasis by regulating the proliferation and differentiation of keratinocytes,release of inflammatory factors,and angiopoiesis.This review summarizes research advances in roles of microRNAs and lncRNAs in psoriasis.
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Objective@#To explore the association of arsenic with unexplained recurrent spontaneous abortion (URSA).@*Methods@#A case-control study was conducted to select URSA patients who were admitted to the Beijing Maternal and Child Health Care Hospital affiliated to Capital Medical University from April to October 2018 as a case group. Women who had a normal pregnancy in the Family Planning Department of the hospital but volunteered to have an abortion were selected as a control group. The case and control group were paired in a 1: 1 ratio. The inclusion criteria of the case group were patients with newly diagnosed recurrent spontaneous abortion who had clinically confirmed more than 2 spontaneous abortions and had 20 weeks prior to pregnancy, excluding patients with recurrent spontaneous abortion caused by abnormal blood coagulation (anti-phospholipid antibody positive), abnormal physiological anatomy (B-ultrasound), abnormal immune factors (anti-nuclear antibody positive, anti-cardiolipin antibody, etc.), genetic chromosomal abnormalities (karyotype analysis) and pathogenic microbial infection. The control group was matched according to the age of the case group (±3 years old) and the gestational age (±2 weeks) to exclude adverse pregnancy outcomes such as stillbirth, congenital malformation, premature delivery and low birth weight infants. A total of 192 subjects were included. Questionnaires were used to collect information of all subjects, and 12 ml of peripheral venous blood was collected to detect blood arsenic levels. Blood arsenic levels were divided into low concentration group (<1.00 μg/L), medium concentration group (1.00-1.50 μg/L) and high concentration group (>1.50 μg/L). The multivariate conditional logistic regression was performed to analyze the relationship between blood arsenic exposure and URSA and explore the influencing factors of blood Arsenic.@*Results@#The geometric mean values of blood arsenic level in the cases group and control group were 1.68 (1.50-1.86) μg/L and 1.26 (1.17-1.37) μg/L, respectively. The blood arsenic level in the case group was significantly higher than that in the control group (P<0.05). The results of multivariate conditional logistic regression analysis showed that after adjusting for tobacco exposure during pregnancy, pre-pregnancy body mass index and the effects of residential decoration in past five years, the risk of URSA was higher in the high-concentration group compared with the low-concentration group (OR=2.56, 95%CI:1.06-6.24).@*Conclusion@#Blood arsenic may increase the risk of URSA in women of childbearing age.
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Objective To compare the efficacy and safety of the long-term intermittent maintenance treatment with tacrolimus 0.03％ ointment versus traditional treatment in reducing relapses and prolonging the recurrence interval in children with moderate to severe atopic dermatitis (AD).Methods A two-phase randomized,open-labelled,controlled clinical trial was conducted from September 2012 to November 2013.In the first phase,a total of 171 children aged 2-15 years with moderate to severe AD were enrolled from 7 hospitals in China,and received conventional treatment with tacrolimus 0.03％ ointment twice a day for 2-6 weeks.At the end of the treatment,the patients who achieved an investigator's global assessment (IGA) score ≤ 2 (n =125) were randomly classified into 2 groups to receive the second-phase treatment:test group (n =62) receiving intermittent maintenance treatment with tacrolimus 0.03％ ointment twice a week (Monday and Thursday),and control group (n =63) receiving no treatment.If the patients in the 2 groups experienced relapse,they received conventional treatment with tacrolimus 0.03％ ointment twice a day.The overall observation period was 6 months.The primary endpoint was the time to the first relapse,which was defined as the number of days from the end of the first-phase treatment to the first relapse.The secondary endpoints included the number of relapses at the second-phase trial,the disease severity at the time of relapse,the duration of relapse,the pruritus score at the time of relapse,the total amount of tacrolimus ointment used,the total response rate at the second-phase trial,and the incidence of adverse events.Results A total of 125 children with AD were enrolled into the second-phase trial,and 121 of them completed the follow-up.Among the 121 patients,the recurrence rate was significantly lower in the test group (25/60,41.7％) than in the control group (46/61,75.4％;x2 =14.20,P ＜ 0.001).The time to the first relapse was significantly longer in the test group (46.9 ± 37.7 d) than in the control group (28.8 ± 32.3 d;Z =1 093.50,P =0.020).The total number of recurrence was 31 and 86 in the test group and control group respectively,and the mean number of recurrence in each patient was significantly lower in the test group (0.52 ± 0.68) than in the control group (1.41 ± 1.23,t =4.96,P ＜ 0.001).There were no significant differences between the two groups regarding disease severity during relapse (eczema area and severity index:Z =971.50,P =0.39),duration of relapse (Z =747.00,P =0.07),and pruritus score during relapse (Z =894.00,P =0.95).The therapeutic drug was tolerated well in all the children,and no tacrolimus-related serious adverse events occurred.Conclusion The intermittent maintenance treatment with tacrolimus 0.03％ ointment twice a week for 6 months can effectively and safely prevent and reduce relapses,and prolong the recurrence interval in children with moderate to severe AD.
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Retrospective analysis of clinical data was conducted in the patients with secondary hyperparathyroidism( SHPT) undergone parathyroidectomy ( PTX ) in our department from September 2015 to September 2016. Uni- and multivariate analyses were conducted to define independent influencing factors for persistent severe hypocalcemia. The results showed that 27 cases(31％ ) suffered from persistent severe hypocalcemia in a total of 87 patients. Univariate analysis revealed that age, weight, hospitalization time after operation, the year of increased serum creatinine, preoperative alkaline phosphatase ( ALP), preoperative intact parathyroid hormone (iPTH), preoperative albumin, postoperative 1 d serum calcium and phosphorus level, renal hypertension, and degeneration of bone were risk factors of persistent severe hypocalcemia. Multivariate analysis revealed that the year of increased serum creatinine, preoperative ALP, and preoperative albumin were the independent influence factors for persistent severe hypocalcemia.
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Objective To evaluate the regulatory effects of lycopene on the key signaling receptors in human cutaneous squamous cell carcinoma cell line COLO16.Methods Cultured COLO16 cells were divided into 6 groups to be treated with lycopene at different concentrations of 0,5,10,15,20,and 25 μmol/L,respectively,for 24 hours (control group and 5,10,15,20,25 μmol/L lycopene groups),followed by estimation of the cell viability by lactate dehydrogenase (LDH) assay.Lycopene at a safe concentration was selected based on the LDH assay,and used for the determination of expression of signaling receptors,and Western blot analysis was performed to measure the expression of key signaling receptor proteins,including epidermal growth factor receptor (EGFR),glucocorticoid receptor (GR),retinoic acid receptor-alpha (RAR-α),retinoid X receptor-alpha (RXR-α),androgen receptor (AR) and progesterone receptor (PR).Statistical analysis was carried out by one-way analysis of variance (ANOVA),Tukey multiple comparison teat and Brown-Forsythe test with the SPSS software.Results After 24-hour treatment with lycopene at different concentrations,there were significant differences in the rate of cell death among these groups (F =13.116,P ＜ 0.05),and the rate of cell death in the 25 μmol/L lycopene group significantly differed from that in the control group (P ＜ 0.05).Therefor,lycopene at concentrations of 5,10 and 20 μmol/L were selected to treat COLO16 and HaCaT cells as well as human epidermal keratinocyte (HEK) for 24 hours in the following experiment.The treatment with lycopene significantly decreased the phosphorylation level of EGFR (P ＜ 0.05),but significantly increased the expression of GR protein (P ＜ 0.05),and showed no significant effects on the protein expression of RAR-α,RXR-α,AR,and PR in COLO16 cells.After 24-hour treatment with lycopene at concentrations of 5,10 and 20 μmol/L,there were no significant changes in the phosphorylation level of EGFR protein or the expression of GR protein in HaCaT cells and HEK (all P ＞ 0.05) compared with those without lycopene treatment.Conclusion Lycopene can decrease the viability of COLO16 cells,inhibit the activation of EGFR protein,and up-regulate the expression of GR,and these effects may be specific for tumor cells.
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Objective To analyze clinical features and treatment outcomes of female children with condyloma acuminatum.Methods Female outpatients with condyloma acuminatum aged less than 12 years were enrolled into this study.Through a questionnaire survey,medical histories were collected,and a physical examination was performed in these patients.Polymerase chain reaction (PCR) was conducted to determine the genotypes of human papillomavirus (HPV).Descriptive statistical methods were used to identify high risk factors for condyloma acuminatum,manifestations of skin lesions,and results of HPV test.The number of sessions of laser therapy and photodynamic therapy was also recorded.Results A total of 51 female children aged under 12 years with a diagnosis of condyloma acuminatum were enrolled into this study.Their median age and course of disease (M [P25-P75]) were 3 (2-5) years and 6 (4-8) months respectively.Of the 51 patients,29 (56.9％) lived in the small towns or suburb of a city,and 16 (31.4％) lived in the rural areas.Only 2 children's parents had a definite HPV infection history.Twelve children (23.6％) usually wore open-seat pants,24 (47％) often took a bath in public bath houses,and only 1 child had a definite history of sexual assault.Among these patients,39 (76.5％) mainly had perianal skin lesions,and all cauliflower-like lesions occurred on the perianal area.A total of 35 patients underwent the HPV test in the exfoliated cells.The positive rate of HPV was 71.4％,and all the HPV-positive patients were infected by low-risk HPV types (HPV6/11).During the treatment,5 patients were lost to follow-up.In the remaining 46 patients,the median number of sessions of laser therapy and photodynamic therapy was 2 (1-4) and 3 (3-4) respectively.The median course of treatment was 4 (2-6) months.After the treatment,all the remaining 46 patients were cured with a recovery rate of 90.2％ (46/51).Conclusions In these female children with condyloma acuminatum,the source of infection may maninly come from the environment,and skin lesions mostly occur on the perianal area.The prognosis is good after laser and photodynamic therapy.
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Objective To investigate molecular mechanisms underlying the synergistic damage to the human squamous cell carcinoma cell line COLO-16 by everolimus and cisplatin.Methods In the signaling pathway experiment,COLO-16 cells were divided into 4 groups:control group receiving no treatment,50,100 and 200 nmol/L everolimus groups treated with 50,100 and 200 nmol/L everolimus respectively.In the combined experiment,COLO-16 cells were divided into another 4 groups:control group,50 nmol/L everolimus group,25 mol/L cisplatin group,and 50 nmol/L everolimus + 25 mol/L cisplatin group.Western blot analysis was performed to analyze changes in mammalian target of rapamycin (mTOR) pathway,Akt pathway,DNA damage-related pathway and Csk homologous kinase (Chk) pathway.Results After the treatment with everolimus at different concentrations of 50,100 and 200 nmol/L for 12 and 24 hours,the phosphorylation levels of mTOR at ser2448 and ser2481 as well as Rictor at thr1 135 in COLO-16 cells were all decreased compared with the control group.However,there were no significant changes in the phosphorylation levels of downstream signals ULK1 at ser757,p70 S6 at thr389 and PKCα at thr638/64.The treatment with everolimus did not change the total protein level and phosphorylation of Akt.After the treatment with cisplatin for 12 and 24 hours,the phosphorylation levels of Rictor at thr1135 and Chk1 at ser345 were significantly increased,but the treatment with everolimus alone showed no such effects.After the combined treatment with everolimus and cisplatin for 12 and 24 hours,the upregulation of Chk1 and Rictor phosphorylation were significantly inhibited compared with the cisplatin alone group.Conclusions mTOR signaling is sensitive to everolimus in COLO-16 cells,but its targeted pathway is not regulated simultaneously to develop a cascade reaction.Everolimus may increase the cisplatin-induced death of COLO-16 cells by inhibiting the activation of Chk 1,but can not aggravate DNA damage induced by cisplatin.
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<p><b>OBJECTIVE</b>To delineate the phenotypic characteristics of 22q11.2 deletion syndrome and the role of CRKL gene in the pathogenesis of cardiac abnormalities.</p><p><b>METHODS</b>G-banded karyotyping, single nucleotide polymorphism (SNP) array and fluorescence in situ hybridization (FISH) were performed on a fetus with tetralogy of Fallot detected by ultrasound. Correlation between the genotype and phenotype was explored after precise mapping of the breakpoints on chromosome 22q11.2. SNP array was also performed on peripheral blood samples from both parents to clarify its origin.</p><p><b>RESULTS</b>The fetus showed a normal karyotype of 46,XY. SNP array performed on fetal blood sample revealed a 749 kb deletion (chr22: 20 716 876-21 465 659) at 22q11.21, which encompassed the CRKL gene but not TBX1, HIRA, COMT and MAPK1. Precise mapping of the breakpoints suggested that the deleted region has overlapped with that of central 22q11.2 deletion syndrome. SNP array analysis of the parental blood samples suggested that the 22q11.21 deletion has a de novo origin. The presence of 22q11.21 deletion in the fetus was also confirmed by FISH analysis.</p><p><b>CONCLUSION</b>Central 22q11.21 deletion probably accounts for the cardiac abnormalities in the fetus, for which the CRKL gene should be considered as an important candidate.</p>
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Adult , Female , Humans , Male , Pregnancy , Adaptor Proteins, Signal Transducing , Genetics , Chromosome Deletion , Chromosomes, Human, Pair 22 , Genetics , DiGeorge Syndrome , Diagnosis , Embryology , Genetics , Fetal Diseases , Diagnosis , Genetics , Genotype , In Situ Hybridization, Fluorescence , Nuclear Proteins , Genetics , Phenotype , Prenatal DiagnosisABSTRACT
<p><b>OBJECTIVE</b>To investigate the value of single nucleotide polymorphism array (SNP array) for the identification of de novo mutations in the DMD gene among fetuses.</p><p><b>METHODS</b>G-banded karyotyping and SNP array were performed on a fetus with intrauterine growth restriction but without family history of Duchenne/Becker muscular dystrophy (DMD/BMD). Multiplex ligation-dependent probe amplification (MLPA) was subsequently applied on amniocytes and maternal peripheral blood sample to detect DMD gene deletion/duplication mutations.</p><p><b>RESULTS</b>Karyotyping of amniocytes showed a normal 46, XY karyotype. SNP array on amniocytes detected a 116 kb deletion (chrX: 32 455 741-32 571 504) at Xp21.1 with breakpoints at introns 16 and 30 respectively, encompassing exons 17-29 of the DMD gene. In addition, MLPA analysis of the DMD gene on amniocytes confirmed the deletion of exons 17 to 29 identified by SNP array. However, no deletion/duplication mutation was detected by MLPA in the mother.</p><p><b>CONCLUSION</b>The de novo deletion of exons 17 to 29 of the DMD gene detected in the fetus may result in BMD or DMD. SNP array can improve the efficiency for detecting genomic disorders in fetuses with unidentified pathogenic genes, negative family history and nonspecific phenotypes.</p>
Subject(s)
Adult , Female , Humans , Pregnancy , Dystrophin , Genetics , Exons , Genetics , Fetus , Congenital Abnormalities , Gene Deletion , Muscular Dystrophy, Duchenne , Genetics , Phenotype , Polymorphism, Single Nucleotide , GeneticsABSTRACT
Objective To evaluate the synergistic effect of everolimus on cisplatin-mediated cytotoxicity against human cutaneous squamous cell carcinoma COLO-16 cells.Methods Cultured COLO-16 cells were divided into several groups to be treated with everolimus at different concentrations of 50,100 and 200 nmol/L or 25 μmol/L cisplatin for 12 and 24 hours.Acridine orange (AO)-labeled autophagic vesicles combined with lysomal enzyme inhibitors (E64d and pepstatin) were used to detect the levels of autophagy and autophagic flow.Western blot analysis was performed to track the conversion of the autophagosome marker microtubule-associated protein 1 light chain-3 (LC3)-Ⅰ to LC3-Ⅱ,as well as to detect cleavage levels of Caspase 3 and poly-ADP-ribose polymerase (PARP).Lactate dehydrogenase (LDH) assay was conducted to detect cell death,and Annexin V-EGFP staining to evaluate cell apoptosis.Results The LC3-Ⅱ / LC3-Ⅰ ratios (LC3-Ⅰ conversion to LC3-Ⅱ) after 12-and 24-hour treatment did not differ among the 50-,100-and 200-nmol/L everolimus groups (12 hours:3.52 ± 0.21 vs.4.03 ± 0.39 vs.5.05 ± 0.22,P ＞ 0.05;24 hours:3.38 ± 0.26 vs.3.29 ± 0.06 vs.6.57 ± 0.16,P ＞ 0.05),but were significantly higher in the three everolimus groups than in the control group receiving no treatment (12 hours:2.07 ± 0.05,P ＜ 0.05;24 hours:2.61 ± 0.16,P ＜ 0.05).After 12-hour treatment,no significant differences were observed in the ratio of LC3-Ⅱ to β-actin between the 50-nmol/L everolimus + E64d + pepstatin group (1.26 ± 0.40),100-nmol/L everolimus ± E64d + pepstatin group (1.16 ± 0.34),200-nmol/L everolimus + E64d + pepstatin group (1.21 ± 0.39) and E64d + pepstatin group (1.19 ± 0.27,P ＞ 0.05).Moreover,there was no significant difference in the percentages of autophagic vesicle-positive cells between the 100-nmol/L everolimus + E64d + pepstatin group and E64d + pepstatin group (2.06％ ± 0.61％ vs.1.68％ ± 0.62％,P ＞ 0.05).After 24-hour treatment,the everolimus + cisplatin group showed significantly increased rate of cell death compared with the cisplatin alone group (42.58％ ± 0.93％ vs.18.20％ ± 1.46％).However,no significant differences were observed in the cleavage levels of Caspase 3 and PARP,the number of annexin V-labelled cells and ratio of LC3-Ⅱ to β-actin between the everolimus + cisplatin group and the cisplatin-alone group (P ＞ 0.05).Conclusion Everolimus has a synergistic effect on the cisplatin-mediated COLO-16 cell death,and this effect does not depend on cell apoptosis or autophagy.
ABSTRACT
Objective To evaluate the efficacy and safety of Qingpeng ointment for the treatment of localized eczema in children.Methods A multicenter,randomized,open-labeled,tacrolimus 0.03％ ointment-controlled clinical trial was conducted.A total of 442 children with localized eczema were enrolled into this study,and divided into 2 groups to topically apply Qingpeng ointment (Qingpeng group) and tacrolimus 0.03％ ointment (tacrolimus group) respectively twice a day for 2 weeks.The children were followed up before the treatment and 1,2 weeks after the treatment,and the efficacy and safety were evaluated at the same time.Results Clinical data from 426 children were included in the efficacy analysis.At 1,2 weeks after the treatment,there were no significant differences in the response rate between the Qingpeng group and tacrolimus group (1 week after the treatment:78.92％ vs.81.77％,x2 =0.545,P =0.460;2 weeks after the treatment:98.65％ vs.97.54％,x2 =0.721,P =0.396).However,the pruritus scores at 1,2 weeks after the treatment were both significantly lower in the Qingpeng group than in the tacrolimus group (1 week:F =14.001,P =0.000;2 weeks:F =11.252,P =0.001).At 1 week after the treatment,the incidence rate of adverse reactions was significantly lower in the Qingpeng group than in the tacrolimus group (P ＜ 0.05).Mild itching,burning sensation and other local irritant sensations were the most common adverse reactions.Conclusion Qingpeng ointment is safe and effective for the treatment of localized eczema in children with good tolerability.