Increased Circulating CXCL10 in Non-Segmental Vitiligo Concomitant with Autoimmune Thyroid Disease and Alopecia Areata

BACKGROUND: Vitiligo is a common acquired pigmentary disease caused by destruction of epidermal melanocytes in underlying autoimmune response. Few studies have been focused on the role of chemokines in non-segmental vitiligo (NSV) concomitant with autoimmune thyroid disease (AITD) and alopecia areata (AA). OBJECTIVE: The aim of this study was to determine the best serum biomarker for predictive role in the progression of vitiligo and to evaluate the influence of AA and/or AITD on vitiligo by using the biomarker. METHODS: This prospective cohort study recruited 45 NSV patients: 14 without either AITD or AA, 12 with AITD, 11 with AA, and 8 with both AITD and AA. Serum levels of CXCL1, CXCL8, CXCL9, CXCL10, CXCL12, CXCL13, and CXCL16 were analyzed by ELISA. CXCR3 mRNA expression was detected on PBMCs by RT-PCR. Improvement was evaluated using repigmentation scales. RESULTS: Serum CXCL10 levels, along with the expression of CXCR3 mRNA were higher in NSV patients with AITD or AA alone than in those without AITD or AA. Moreover, serum CXCL10 levels, along with the expression of CXCR3 mRNA were higher in NSV patients with both AITD and AA than in those with AITD or AA alone. Poorer repigmentation was observed in NSV patients with both AA and AITD than in those with AA or AITD alone. CONCLUSION: CXCL10 could be a biomarker to predict the progression of NSV. Dermatologists should pay much attention to those NSV patients concomitant with AITD and/or AA, for comorbidity might lead to more active autoimmune reaction.

Regulatory role of cathepsin D in degradation of advanced glycation end products by human dermal fibroblasts / 中华皮肤科杂志

Objective To investigate the regulatory role of cathepsin D (CatD) in the degradation of intracellular advanced glycation end products (AGEs) endocytosed by human dermal fibroblasts (HDFs).Methods Cultured HDFs were treated with 1 μnol/L CA074Me (an inhibitor of CatB and CatL),75 μmol/L pepstatin A (an inhibitor of CatD) and 1 μmol/L MG-132 (an inhibitor of20S proteasome) separately for 4 hours,and then cell counting kit 8 (CCKS) assay and fluorometric assay were performed to determine the cellular viability and protease activity,respectively.The cells in the CA074Me group,pepstatin A group and MG-132 group were additionally treated with AGE-bovine serum albumin (BSA) for 8 hours,and the cells in the blank control group were treated with phosphate-buffered saline (PBS) alone.After 8-hour cultivation,the cells in the above groups were subsequently reincubated with fresh culture medium containing the corresponding inhibitors for 24 hours.Then,flow cytometry was performed to assess the mean fluorescence intensity of intracellular AGE-BSA at different time points.Some other HDFs were treated with 37.5,75 and 150 μmol/L pepstatin A and PBS separately for 4 hours,and then the cells in the 4 groups were treated with 200 mg/L AGE-BSA for 8 hours,followed by the removal of AGE-BSA from the medium and the treatment with 37.5,75 and 150 μmol/L pepstatin A and PBS respectively.Enzyme-linked immunosorbent assay (ELISA) was conducted to measure the mean concentration of intracellular AGE-BSA at different time points,and the degradation rate of AGE was calculated.Some HDFs were divided into 3 groups:blank control group receiving no treatment,NC group transfected with an empty vector,and CatD group transfected with a CatD-overexpressing lentiviral vector.Fluorescence microscopy was conducted to estimate the transfection efficiency.Reverse transcription-PCR,Western blot analysis and fluorometric assay were performed to determine the mRNA and protein expression,and activity of CatD respectively.Then,the cells in the above 3 groups were incubated with AGE-BSA for 8 hours,followed by the removal of AGE-BSA from the medium and the treatment with fresh culture medium.The detection methods were same as the above experiment,and the degradation rate was calculated.Results The cellular proliferative activity in the 1-μmol/L CA074Me group,75-μmnol/L pepstatin A group and 1-μ mol/L MG-132 group was more than 90％,and there was no significant difference between the 3 groups and the control group (100％,F =1.525,P ＞ 0.05).Twenty-four hours after the removal of AGE-BSA from the medium,the fluorescence intensities of intracellular AGE-BSA in the CA074Me + AGE-BSA group (275.00 ± 10.15) and MG-132 + AGE-BSA group (259.00 ± 11.14) significantly decreased compared with those at the 8-hour time point (295.00 ± 6.56 and 285.67±8.74 respectively;paired t test,t =4.778,6.154 respectively,both P ＜ 0.05),while no significant difference was observed in the fluorescence intensities of intracellular AGE-BSA in the pepstatin A + AGE-BSA group between the 8-hour time point and 32-hour time point (P ＞ 0.05).The degradation rates of intracellular AGE-BSA within 24 hours in the 37.5,75 and 150 μmol/L pepstatin A groups were 9.64％ ± 1.27％,5.62％ ± 0.47％ and 3.21％ ± 0.73％ respectively;there were significant differences among the 3 groups (F =45.876,P ＜ 0.05),and the degradation rate significantly decreased along with the increase of pepstatin A concentration (P ＜ 0.05).Fluorescence microscopy showed no fluorescent cells in the blank control group,while the NC group and CatD group both showed a high proportion (＞ 80％) of fluorescent cells.The mRNA and protein expression as well as the activity of CatD were significantly higher in the CatD group than in the blank control group and NC group (all P ＜ 0.05).The CatD + AGE-BSA group showed a significantly higher degradation rate of intracellular AGE-BSA within 24 hours compared with the AGE-BSA group and NC + AGE-BSA group (both P ＜ 0.05).Conclusion CatD can promote the degradation of intracellular AGE-BSA endocytosed by HDFs.

Correlation analysis between the expression of cathepsin D and the accumulation of advanced glycation end products in skin tissues / 中华皮肤科杂志

Objective To determine the expression of cathepsin D and advanced glycation end products (AGEs)in skin tissues from patients of different ages or skin tissues with different degrees of sun exposure,to evaluate their correlation,and to preliminarily investigate the role of cathepsin D in the degradation and accumulation of AGEs in photoaged skin.Methods Skin tissues were collected from sunexposed and sun-protected body sites in patients aged 15-20 years,35-40 years,55-60 years or 75-80 years.These skin tissues were divided into 8 groups according to age of patients and degrees of sun exposure,and there were 6 specimens in each group.Immunohistochemical and immunofluorescent methods were used to measure the expression of cathepsin D and AGEs in the skin tissues.Statistical analysis was carried out by factorial design analysis of variance,Wilcoxon rank sum test and Kruskal-Wallis rank sum test for analyzing associations of the expression of cathepsin D and AGEs with age and sun exposure,as well as by Pearson correlation analysis for assessing the correlation between cathepsin D expression and AGEs expression.Results Immunohistochemical study showed that the expression of cathepsin D markedly decreased along with the increase of age,but the accumulation of AGEs gradually increased along with the increase of age.In the same age group,the cathepsin D expression was lower in the sun-exposed skin tissues than in the sun-protected skin tissues,while the accumulation of AGEs was more in the sun-exposed skin tissues than in the sun-protected skin tissues.Factorial design analysis of variance showed that sun exposure could decrease the expression of cathepsin D (F =58.70,P ＜ 0.001),but increase the accumulation of AGEs (F =158.18,P ＜ 0.001).Moreover,the increase of age could lead to decreased expression of cathepsin D (F =79.49,P ＜ 0.001),and increased expression of AGEs (F =106.06,P ＜0.001).Compared with the sun-protected skin tissues,the sun-exposed skin tissues in all the age groups showed significantly lower absorbance value of cathepsin D (35-40 years:0.020 ± 0.005 vs.0.032 ± 0.005;55-60 years:0.012 ± 0.004 vs.0.026 ± 0.002;75-80 years:0.002 ± 0.001 vs.0.013 ± 0.004;all P ＜0.001),but higher absorbance value of AGEs (35-40 years:0.030 ± 0.008 vs.0.010 ± 0.003;55-60years:0.066 ± 0.010 vs.0.021 ± 0.004;75-80 years:0.085 ± 0.015 vs.0.035 ± 0.009;all P ＜ 0.001)except the age group of 15-20 years.No matter whether the skin tissues were sun-exposed or sunprotected,there were significant differences in the expression of cathepsin D and AGEs among different age groups (all P ＜ 0.001).The results of double immunofluorescence staining were similar to those of immunohistochemical study.Pearson correlation analysis showed that the expression of cathepsin D in the sun-exposed skin tissues was highly negatively correlated with the accumulation of AGEs (r =-0.915,P ＜0.05),while they were moderately negatively correlated in the sun-protected skin tissues (r =-0.730,P ＜0.05).Conclusions Along with the increase of age,the expression of cathepsin D in skin tissues decreased,but the expression of AGEs increased.In the sun-protected skin tissues,the expression of cathepsin D was moderately negatively correlated with the expression of AGEs,while they were highly negatively correlated in the sun-exposed skin tissues,suggesting that cathepsin D may play an important role in the degradation and accumulation of AGEs in photoaged skin.

Effect of photoaging on the degradation of advanced glycation end products by human dermal fibroblasts / 中华皮肤科杂志

Objective To evaluate the effect of photoaging on the degradation of advanced glycation end products (AGEs) by human dermal fibroblasts.Methods Some cultured human dermal fibroblasts were subjected to repetitive ultraviolet A (UVA) radiation (UVA radiation group) to establish a photoaging cell model,which was then evaluated by cell counting kit 8 (CCK-8) assay,senescenceassociated β-galactosidase staining and detection of apoptosis rate.Moreover,fibroblasts receiving no treatment served as control group.Some other primary fibroblasts were divided into 4 groups:photoaged group receiving UVA radiation,non-photoaged group receiving no treatment,AGE-treated photoaged group treated with UVA radiation followed by the treatment with 200 mg/L AGE-bovine serum albumin (BSA),and AGE-treated non-photoaged group treated with 200 mg/L AGE-BSA alone.After the treatment with AGE-BSA for 4-72 hours,flow cytometry was performed to determine the fluorescence intensity of AGE-BSA in fibroblasts of the above groups.After 8-hour treatment with AGE-BSA,confocal laser scanning microscopy was performed to localize and semiquantitatively detect AGE-BSA in fibroblasts,and enzymelinked immunosorbent assay (ELISA) was conducted to detect AGE-BSA levels in fibroblasts,as well as changes in the intracellular AGE-BSA level within 24 hours after the removal of AGE-BSA.Results Compared with the control group,the UVA radiation group showed significantly decreased cellular proliferative activity (t =7.559,P ＜ 0.05),but significantly increased apoptosis rate and percentage of β-galactosidase-positive fibroblasts (t =14.075,43.524 respectively,both P ＜ 0.05).Flow cytometry revealed that the average fluorescence intensities of AGE-BSA after 4-,8-,16-,24-,48-and 72-hour treatment with AGE-BSA were significantly higher in the AGE-treated photoaged group (293.00 ± 8.19,359.67 ± 11.59,347.00 ± 12.29,338.00 ± 12.77,334.67 ± 14.22 and 336.30 ± 10.21,respectively) than in the photoaged group (all P ＜ 0.05),as well as in the AGE-treated non-photoaged group (222.33 ± 8.74,276.33 ± 6.11,256.33 ± 5.51,243.00 ± 10.15,236.33 ± 1.53 and 240.33 ± 1.52,respectively) than in the non-photoaged group (all P ＜ 0.05).Moreover,the average fluorescence intensities of AGE-BSA at different time points were all significantly higher in the AGE-treated photoaged group than in the AGE-treated non-photoaged group (all P ＜ 0.05).Confocal laser scanning microscopy showed that AGE-BSA was mainly localized in lysosomes after endocytic uptake into the fibroblasts,and the AGE-treated photoaged group showed significantly increased fluorescence intensity of AGE-BSA compared with the AGE-treated non-photoaged group (P ＜ 0.05).ELISA revealed that the intracellular AGE level in the AGE-treated non-photoaged group at 24 hours after the removal of AGE-BSA was decreased by (14.6 ± 1.2)％ compared with that before the removal,and the degradation rate of AGE-BSA was significantly higher in the AGE-treated non-photoaged group than in the AGE-treated photoaged group (7.6％ ± 1.4％,t =6.604,P ＜ 0.05).Conclusion The internalized AGE-degradating ability decreases in photoaged fibroblasts,which may induce the accumulation of AGEs in photoaged skin.

Effect analysis of nursing intervention on patients with recurrent abortion treated with progesterone / 中国生化药物杂志

Objective To investigate recurrent spontaneous abortion patients received bends the clinical application effect of nursing intervention during progesterone treatment. Methods Two patients with recurrent spontaneous abortion group were treated with progesterone treatment, the control group in the conventional nursing service only provides flexor progesterone treatment on the basis of comprehensive nursing service study group progesterone treatment on the basis of routine the combination of nursing and nursing intervention in the 2 groups of patients with recurrent spontaneous abortion. The success rate of pregnancy were recorded, the rate of abortion, the data will be given the statistical analysis (using SPSS software) after the conclusion. Results The study group of patients with recurrent spontaneous abortion pregnancy success rate (95.45%) and control group (90.91%) there is no significant difference between the study group of patients with recurrent spontaneous abortion abortion rate (6.82%) was significantly lower than the control group (34.09%), there was significant difference between the groups, P<0.05. Conclusion Recurrence During the treatment of flexor progesterone abortion were given routine nursing and nursing intervention combined with the comprehensive clinical nursing service can obtain ideal success rate of pregnancy, to reduce the rate of abortion is also of positive significance, is conducive to the protection of patients' quality of life, physical and mental health.

Effect of Fukeqianjin tablets on uterine bleeding patients serum immunoglobulin IgM, IgA, IL-2 in serum and its soluble receptor function / 中国生化药物杂志

Objective To investigate the effect of Fukeqianjin tablets combined with progesterone on serum immunoglobulin,serum IL-2 and soluble receptor levels in patients with functional uterine bleeding,and to explore the optimal regimen for patients with functional uterine bleeding.Methods A total of 98 patients with functional uterine bleeding diagnosed in our hospital from June,2015 to June,2016 were randomLy divided into observation group and control group(n=49).The patients in the control group were treated with oral progesterone and the observation group.The control group was treated on the basis of the combination of gynecological Qianjin Tablet,compared the two groups of patients before and after treatment of menstrual conditions,serum IgM,IgA,serum IL-2 and soluble receptor levels.Results In the first month,the second month and the third month after treatment,the menstrual volume in the observation group were(103.5±21.5)mL,(93.5±14.7)mL,(81.4± 12.2)(7.68±0.92)d,(6.81±0.87)d and(6.14±0.78)d respectively in the observation group were significantly shorter than those in the control group(P<0.05).After treatment,the menstrual period The serum levels of IgA and IgM were(3.6±0.8),(2.6±0.8),(1.8±0.3)and(2.9±0.5),(2.2±0.3),(1.1±0.2)in the first month,the second month and the third month respectively,Significantly lower than the control group,the difference was statistically significant; the first month after treatment,the first two months,the first three months,observation group of patients with serum IL-2 and soluble receptor levels significantly Lower than the control group.Conclusion Fukeqianjin tablets combined with progesterone can improve the immune function of patients with functional uterine bleeding,reduce the level of inflammation and soluble receptor levels,compared with single progesterone treatment better,the program better.

Effects of different doses of dexmedetomidine on etomidate in inhibiting the EC50 of laryngeal mask placement / 中国生化药物杂志

Objective To investigatethe EC50 of different doses of dexmedetomidine on etomidate inhibited responses tolaryngeal maskinsertion in patients.Methods 88 with breast cancerfrom surgical department in Zhuji Hospital of Traditional Chinese Medical from August 2014 to August 2015were selected and randomly divided into the control group(groupA)and the experiment group(group B1,group B2 and group B3)with 22 casesin each group,respectively,intravenous pump 0.9％sodium chloride solution and DEX(dose followed by 0.3,0.6,0.9μg/kg).The next sequential intravenous infusion of etomidateafter 10 min.The EC50 and the 95％confidence interval of etomidate were determined by sequential method in each group of patients,the changes of vital signs and adverse reactionsin patients were monitored.Results Four groups of patients with LMA EC50 and 95％confidence interval of etomidate respectively:0.78(0.723～0.835)μg/mL in group A,0.66(0.612～0.711)μg/mL in group B1,0.58(0.532～0.627)μg/mL in group B2,0.46(0.416～0.521)μg/mL in group B3.Four groups of patients with laryngeal mask insertion immediately before MAP and HR were lower than the baseline value,the difference was statistically significant(P<0.05),but the LMA elevated after one min.The control group of four patients with respiratory depression,three cases of patients with bradycardia in group B3,two cases of patients with hypotension were improved after symptomatic treatment.Conclusion In a certain range,increasing the dose of dexmedetomidine reduces the effective concentration of etomidate,which inhibits the laryngeal mask placement reaction.

Effect of lentinan injection in different treatment administration of oxaliplatin combined with capecitabine in gastric cancer and the effect on the peripheral blood of patients with CEA and CA199 levels / 中国生化药物杂志

Objective To investigate the effect of lentinan injection in different treatment administration of oxaliplatin combined with capecitabine in treatment of gastric cancer and the effect on the peripheral blood of patients with CEA and CA199 levels.Methods 90 cases in our hospital from September 2009 to September 2012 were selected as research subjects, according to patients with lentinan injection treatment, the patients were divided into A, B and C three groups, the effect and the adverse effects of chemotherapy in three groups of patients, detection of serum CEA and CA-199 levels. Results CR, PR, SD and PD of A groups were 12, 7, 8, 3 cases, RR was 63.3%, obviously higher than that of B, C two groups, the difference was statistically significant (P<0.05), group A patients with leukopenia I,II and III degree were 15, 10, 5 cases of gastrointestinal reaction, I, II, III were 12, 10, 8 cases of abnormal liver function, I, II and III degree were 14, 9, 7 cases.A group, leukopenia, gastrointestinal tract, liver function abnormal lesion was lower than the B group and C group, the difference was statistically significant (P<0.05), CEA, CA199 level before treatment compared to no difference, one course of chemotherapy, after two courses of A, B, C three groups of patients with CEA and CA199 level were before treatment decreased, and the level of group A was significantly lower than that of B, C two B, C group, no significant difference between the two groups. Conclusion Lentinan injection helps to improve the sensitivity of chemotherapy in patients with gastric cancer, reduce adverse drug reactions, reduce the level of tumor markers, improve the prognosis of patients with gastric cancer.

Effects of advanced glycation end products on the expressions and activity of cathepsin D in ultraviolet A- irradiated human dermal fibroblasts / 中华皮肤科杂志

Objective To investigate the effects of advanced glycation end products(AGE)on the expressions and activity of cathepsin D(CatD)in ultraviolet A(UVA)?irradiated human dermal fibroblasts. Methods Human dermal fibroblasts were isolated and harvested from the circumcised foreskin of children, and subjected to a primary culture. CCK?8 assay was performed to screen non?cytotoxic concentrations of AGE?bovine serum albumin (BSA). Some fibroblasts were incubated with 50, 100 and 300 mg/L AGE?BSA separately for 24 hours, with untreated cells as the control group. Then, reverse transcription(RT)?PCR, Western?blot analysis and a fluorimetric assay were performed to measure the mRNA and protein expressions as well as activity of CatD, respectively. Some fibroblasts were classified into six groups: control group receiving no treatment, AGE?BSA group and BSA group treated with the highest non?cytotoxic concentration of AGE?BSA and the same concentration of BSA respectively for 24 hours, UVA group irradiated by 10 J/cm2 UVA, UVA?AGE?BSA group and UVA?BSA group treated with AGE?BSA and BSA at the above non?cytotoxic concentration respectively for 24 hours both before and after UVA radiation at 10 J/cm2. After the treatments, RT?PCR, Western?blot analysis and a fluorimetric assay were conducted to detect mRNA and protein expressions and activity of CatD respectively. Results AGE?BSA of 50- 200 mg/L exhibited no obvious influence on cellular proliferation of fibroblasts. The fibroblasts incubated with AGE?BSA of 50, 100 and 200 mg/L showed a significant increase in the mRNA expression(0.267 ± 0.007, 0.348 ± 0.007, and 0.418 ± 0.006 respectively), protein expression (1.403 ± 0.181, 2.233 ± 0.090 and 2.477 ± 0.111 respectively), and activity(1.760 ± 0.080, 2.330 ± 0.060 and 2.890 ± 0.080 respectively)of CatD compared with the control group(mRNA:0.161 ± 0.006;protein:0.903 ± 0.200;activity:1.100 ± 0.090, all P < 0.05). AGE?BSA increased CatD expressions and activity in a dose?dependent manner. The mRNA and protein expressions as well as activity of CatD were significantly higher in the UVA group than in the control group (mRNA expression: 0.480 ± 0.005 vs. 0.155 ± 0.005; protein expression: 2.583 ± 0.199 vs. 0.920 ± 0.235;activity:2.970 ± 0.110 vs. 1.110 ± 0.040, all P<0.05), but significantly lower in the UVA?AGE?BSA group than in the UVA group(mRNA expression:0.394 ± 0.008 vs. 0.480 ± 0.005;protein expression:2.070 ± 0.125 vs. 2.583 ± 0.199;activity: 2.560 ± 0.060 vs. 2.970 ± 0.110, all P < 0.05). Conclusion AGEs could increase CatD expressions and activity in human dermal fibroblasts not receiving UVA irradiation, but inhibit their increase in UVA?induced human dermal fibroblasts.

Effect of ShenQi FuZheng injection on peripheral blood AngⅡ, ET-1 levels and clinical curative effect in patients with congestive heart failure / 中国生化药物杂志

Objective To analysis effect of ShenQi FuZheng injection on peripheral blood angiotensin II(AngⅡ), endothelin-1(ET-1) levels and clinical curative effect in patients with congestive heart failure.Methods 52 patients who were diagnosed with chronic heart failure were collected.All patients were randomly divided into experimental group and control group, 26 cases in each group.Patients in the control group received conventional western medicine treatment, patients in the experimental group were given ShenQi FuZheng injection on the basis of control group treatment, after the treatment, the plasma levels of AngⅡ and ET-1, cardiac function and clinical efficacy were detected in all patients.Results After treatment, compared with control group, the plasma levels of AngⅡ was lower in the experimental group(P<0.05);the plasma levels of ET-1 was lower in the experimental group(P<0.05); the LVEF,CO and SV levels were higher,and the LVED level was lower in the experimental group(P<0.05); the effective rate of patients in the experimental group was higher(P<0.05).Conclusion The ShenQi FuZheng injection can significantly reduce the plasma AngⅡand ET-1 levels in patients with congestive heart failure, improve heart function and clinical curative effect.