Stilbene glycoside (TSG) has been shown to have many beneficial properties. It is therefore essential to understand the absorption and metabolism of TSG in detail. We determined the recovery of TSG and its metabolites (TSG sulfate/glucuronides) in rat gastric contents, gastric mucosa, portal vein plasma, celiac arterial plasma, bile, and urine after administration of 15mg of TSG in 0.5mL physiological saline or incubation for 20min in situ in the stomach of rats. Within 20min, (64.0±9.8)% of the administered TSG disappeared from the stomach; later, TSG was recovered in both free and conjugated forms in plasma and bile, but not in urine. On the other hand, only free TSG was detected in the gastric contents and mucosa; it was also detected in the portal vein plasma as (48.1±3.5)% of the total TSG (all forms of TSG). However, the proportion of free TSG in the celiac arterial plasma and bile decreased to 4%-10%. In addition, the proportion of free TSG to total TSG in the liver microsome incubation mixture after TSG was incubated in liver microsome at 37℃ for 30min was very low [(10.6 ± 2.6)%]. These results indicate that TSG could be quickly absorbed from the rat stomach, conjugated in liver and excreted in bile. Such novel information would be helpful for the use of TSG as a beneficial natural product which may improve its proposed efficacy in preventing chronic diseases.

Objective To investigate the incidence, clinical patterns, and species distribution of pathogenic fungi of onychomyeosis in patients with chronic viral hepatitis (CVH), and to analyze the relationship between onychomycosis and CVH. Methods From November 2005 to October 2006, direct microscopy and fungal culture were performed on nail samples from CVH patients with clinically suspected onychomycosis in the two largest institutions for communicable disease control in Guangzhou city. The incidence, clinical patterns, and species distribution of pathogenic fungi of onychomycosis were assessed based on the findings in mycologic examinations. Results The study randomly recruited 995 patients with CVH, and onychomycosis was diagnosed in 116 patients. The incidence of onychomycosis was 11.66% in total, 6.20%, 8.59%, 14.09%, 19.67% in patients with mild, moderate, severe and extremely severe CVH respectively, 7.09%, 17.29%, 19.13% and 27.27% in patients with a clinical course of CVH of 0.5-9 years, 10-19 years, 20-29 years, ≥30 years respectively. The most common clinical pattern was distal and lateral subungual onychomycosis (DLSO, 69.83%), followed by total dystrophic onychomycosis (TDO, 14.66%). Among the pathogenic fungi, dermatophytes amounted to 71.43%, yeasts 21.43%, moulds 7.14%, and Trichophyton rubrum was the most frequently isolated fungus (42.86%). Conclusions The incidence of onychomycosis in patients with CVH is correlated with the severity and course of CVH. Among these patients, the most common clinical pattern is DLSO with the most frequent fungal species being dermatophytes and predominant fungal isolate being Trichophyton rubrum.

Objective To elucidate the molecular basis for induced resistance of N. gonorrhoeae to ceftriaxone in vitro. Methods The reference strain ATCC49226 and clinical isolate ZSSY00205 of N. gon-orrhoeae were exposed to subinhibitory concentration of ceftriaxone for the induction of resistance. Then,suppression subtractive hybridization was performed with the pre-induction parent strains as drivers and post-induction mutant strains as testers to create a subtractive cDNA library. Following that, a total of 192 clones were randomly selected from the library, and arrayed by spotting onto nylon membranes. Finally, dif-ferentially expressed genes were screened by hybridization with labeled-RsaI restriction fragments from the sensitive and resistant N.gonorrhoeae strains respectively, and analyzed by sequencing and homology research using Blast program. Results A subtractive library for these resistant N.gonorrhoeae strains was generated by SSH technique. Microarray analysis and homology research confirmed 5 genes related to ceftriaxone resistance, i.e. mtrR, mtrC, gyrB, rpsJ and PJD1. Conclusions The induced resistance of N. gonorrhoeae to ceftriaxone may be associated with mtrR, mtrC, gyrB, rpsJ and PJD1 genes which probably mediate the resistance by enhancing the activity of efflux pump system.

Objective To explore the relationship between interleukin 1 receptor accessory protein-like 2 (IL1RAPL2) gene and cognitive abilities of children in Qin-Ba mountainous region. Methods Four tagged SNPs (rs5962434,rs5916817, rs3764765 and rs5962298 ) in IL1RAPL2 were selected, and then genotyped by PCR-SSCP method in a 320 children sample aged from six to fourteen years old. Results The results showed the rs5962434, rs5916817 ,rs3764765 and rs5962298 had no deviations from Hardy-Weinberg equilibrium (P＞0.05),and there were no significant statistical differences in the average psychometric scores of general cognitive ability(P=0.81,0.53,0.79,0.90) ,verbal comprehension (P=0.58,0.47,0.69,0.87 ) ,memory and concentration (P=0.69,0.35,0.76,0.90) among the compared genotype groups at each of the markers. Furthermore,the results also indicated that the four SNPs were not associated with perceptual organization in males and females respectively (P = 0.70,0.85,0.76,0.90,0.65,0.22,0.98,0.90 ). Conclusion The present work suggests that the human general cognitive ability, the three cognitive factors of C-WISC scale are not influenced manifestly by the genetic variations in IL1RAPL2.

Objective To evaluate effectiveness of peripheral Mohs micrographic surgery for the treatment of extramammary Paget′s disease (EMPD). Methods A total of 28 patients with EMPD were treated with peripheral Mohs micrographic surgery. The depth and extent of tumor infiltration were evaluated before the surgery. One day before the surgery, 20% aminolevulinic acid hydrochloride was topically applied to determine and label surgical margins under a Wood′s lamp. After fluorescence-based localization, peritumoral skin tissues were resected and underwent frozen-section examination according to the protocol for Mohs micrographic surgery. Meanwhile, the tumor was resected. After surgery, patients were followed up every 3 - 6 months to detect local recurrence and metastasis. Results Of the 28 patients, 25 were male and 3 were female. Six patients each underwent 3 sessions of frozen-section examination, and 12 patients each received 2 sessions, with an average of 1.86 sessions for each patient. During the follow-up for 5 - 72 months, local recurrence occurred in 3 cases, and 1 patient died of tumor metastasis and uremia after 2 years of follow-up. Conclusion Peripheral Mohs micrographic surgery is a time-saving and effective treatment for EMPD.

Objective To observe the expression changes of cathepsin K (CatK) in human dermal fibroblasts at different time points after different doses of ultraviolet A (UVA) irradiation.Methods Dermal fibroblasts were isolated from circumcised foreskins of children,and subjected to primary culture and subculture.Cells at third-tenth passage were used in the following experiment.Some fibroblasts were irradiated with UVA of 10 J/cm2 and collected at 24,48 and 72 hours separately after the irradiation,and some fibroblasts were irradiated with UVA of 10,20 and 30 J/cm2 separately and harvested 48 hours later.The fibroblasts receiving no irradiation served as the control group.Reverse transcription PCR and Western blot were carried out to detect the mRNA and protein expressions of CatK in fibroblasts,respectively.Results Compared with the control fibroblasts,those irradiated with UVA of 10 J/cm2 showed a significant elevation in the mRNA and protein expression levels of CatK on day 1 (0.351 ± 0.038 vs.0.177 ± 0.006,1.76 ± 0.27 vs.0.82 ± 0.45,respectively,both P＜ 0.05),day 2 (0.510 ± 0.017 vs.0.176 ± 0.002,2.97 ± 0.36 vs.1.58 ± 0.15,respectively,both P＜ 0.05) and day 3 (0.313 ± 0.012 vs.0.173 ± 0.002,2.23 ± 0.14 vs.1.29 ± 0.32,respectively,both P ＜ 0.05),with the highest expressions of CatK mRNA and protein observed on day 2.Within the range of 10-30 J/cm2,UVA enhanced the CatK mRNA and protein expression levels in a dose-dependent manner.In detail,at 48 hours after the irradiation with UVA of 10,20 and 30 J/cm2,the CatK mRNA expression level in the irradiated fibroblasts was 2.34,2.91 and 3.18 times,and the CatK protein expression level 1.77,2.82 and 3.64 times,respectively,that in the control fibroblasts (all P ＜ 0.05).Conclusion The expression of CatK is up-regulated in human dermal fibroblasts after UVA irradiation.

Objective To investigate whether ultraviolet A UVA)-induced CatK expression is regulated by the mitogen-activated protein kinases (MAPK) signaling pathway in human dermal fibroblasts in vitro.Methods Human dermal fibroblasts were obtained from circumcised foreskin of children,and subjected to primary culture.After several passages of subculture,some fibroblasts were irradiated with UVA at a dose of 10 J/cm2.Western blot was performed to measure the expressions of total and phosphorylated JNK (t-and p-JNK) and P38 (t-and p-P38) at 0.75,1.5,3 and 6 hours after the irradiation.Some fibroblasts were divided into six groups:control group receiving no treatment,SP group treated with SP600125 of 800 nmol/L,SB group treated with SB203580 of 10 μmol/L,UVA group irradiated with UVA at a dose of 10 J/cm2,UVA-SP group treated with SP600125 for 1 hour before and for 1.5 or 48 hours after UVA irradiation at 10 J/cm2,UVA-SB group treated with SB203580 for 1 hour before and for 1.5 or 48 hours after UVA radiation at 10 J/cm2.Subsequently,Western blot was performed to determine the expressions of p-c-Jun and p-MAPKAPK2 in these groups at 1.5 hours after the UVA irradiation,and reverse transcription (RT)-PCR and Western blot to detect the mRNA and protein expressions of CatK at 48 hours after the UVA irradiation,respectively.Statistical analysis was carried out by t test,one way analysis of variance and least significant difference (LSD)-t test.Results The expression levels (gray values) of p-JNK and p-P38 were significantly increased at 0.75 hour (4.77 ± 0.19 and 2.44 ± 0.13 respectively,both P ＜ 0.05) and 1.5 hours (4.68 ± 0.09 and 2.30 ± 0.04 respectively,both P ＜ 0.05),but showed no significant changes at 3 hours (both P ＞ 0.05) and 6 hours (both P ＞ 0.05) after the UVA irradiation compared with those before the irradiation (3.2 ± 0.27 and 1.61 ± 0.08 respectively).A significant decrease was observed in the expression of p-c-Jun in the UVA-SP group and p-MAPKAPK2 in the UVA-SB group compared with the UVA group (p-c-Jun,2.55 ± 0.48 vs.4.85 ±0.96; p-MAPKAPK2,1.16 ± 0.12 vs.2.46 ± 0.09,both P ＜ 0.05).The CatK mRNA and protein expressions were attenuated by 61.1％ and 44.3％ respectively in the UVA-SP group (both P ＜ 0.05),and by 71.3％ and 50.4％ respectively in the UVA-SB group (both P ＜ 0.05) in comparison with the UVA group.The UVA-SP group also showed a significant reduction in CatK mRNA and protein expressions as compared with the UVA-SB group (both P ＜ 0.05).Conclusion Both JNK and P38 signaling pathways,especially the JNK pathway,may contribute to the upregulation of CatK expression in dermal fibroblasts induced by UVA irradiation.

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.

Objective:To investigate the effect and mechanism of short-chain fatty acids (SCFAs) on the side-effect of tacrolimus on blood glucose.Methods:The C57BL/6 mice were treated with tacrolimus orally (10 mg/kg, tacrolimus group), tacrolimus plus 150 mmol/L sodium butyrate and isovalerate mixed solution (SCFAs group), broad-spectrum antibiotics (antibiotic group), and tacrolimus plus broad-spectrum antibiotics (tac&abx group). After 8 weeks intervention, the fasting blood glucose (FBG), oral glucose tolerance test (OGTT), hemoglobin A1C (HbA1c) were tested as indicators of glucose metabolism, and the gut microbiota, SCFAs concentration in the ileocecal, serum glucagon-like peptide-1 (GLP-1), fasting serum insulin, and GLP-1 expression in intestinal mucosa were performed for intestinal-glucose metabolism mechanism.Results:The FBG and HbA1c were significantly increased in tacrolimus group[(7.31±0.97)mmol/L, (8.34±1.12)%] than control group [(5.23±0.30)mmol/L, (4.32±0.80)%, all P<0.05], which remained normal in antibiotic group [(4.92±0.31)mmol/L, (5.61±0.98)%)], tac&abx group[(5.95±0.37)mmol/L, (4.56±0.26)%] and SCFAs groups [(5.87±0.68)mmol/L, (5.07±1.79)%]. The OGTT in the tacrolimus group showed glucose tolerance impairment, while other groups remained normal. The ileocecal butyric acid and isovaleric acid concentrations in the tacrolimus group were (722.3±262.2) μg/g and (10.0±5.1)μg/g, lower than the control group[ (1 321.3±165.5) μg/g, (19.7±3.6)μg/g, P<0.05]. The above acids in the SCFAs group remained normal as in the control group [(1 375.7±451.6) μg/g, (24.5±11.5)μg/g)]. The fasting serum insulin in the tacrolimus group decreased significantly to (3.2 ± 0.6)mIU/L, compared with control[ (4.4±0.9) mIU/L]and SCFAs groups [(7.0±1.1) mIU/L]. The GLP-1 test indicated a significant decrease in the tacrolimus group[ (4.7±2.9)pg/ml, P<0.05] compared with the SCFAs group and control group [(42.5±19.9) pg/ml, (33.1±9.1) pg/ml]. Conclusions:Tacrolimus affects glucose metabolism through the SCFAs-associated GLP-1 pathway in the intestine, and oral supplementation with mixed SCFAs provides a new insight for the prevention and treatment of tacrolimus-induced hyperglycemia in transplant recipients.

Objective:To compare the detection rate of genital Chlamydia trachomatis (CT) DNA between urine and urethral/cervical swab samples. Methods:From December 2018 to December 2019, a total of 1 475 outpatients were collected from sexually transmitted disease clinics in 7 medical institutions, such as Department of Venereology, Guangzhou Institute of Dermatology, including 1 118 males and 357 females. One urethral/cervical swab sample and one urine sample were collected successively from each patient. Real-time fluorescence-based PCR was performed to detect CT DNA in urine and urethral/cervical swab samples, and paired chi-square test was used to compare the positive rate of CT DNA between the 2 kinds of samples. Random- or fixed-effect meta-analysis was conducted for the test of heterogeneity and merging of positive rates of CT DNA in the urine and urethral/cervical swabs among 7 medical institutions.Results:The positive rate of CT DNA in the urine samples was significantly higher than that in the swab samples from 4 medical institutions (all P < 0.05) , while there was no significant difference in the positive rate of CT DNA between the 2 kinds of samples from 3 medical institutions (all P > 0.05) . The heterogeneity ( I2) estimates of the CT-DNA positive rate in urine and swab samples among different medical institutions were 78.6% (95% CI: 55.9% - 89.6%) and 73.7% (95% CI: 43.7% - 87.7%) , respectively; meta-analysis showed that the total merged positive rate of CT DNA in the urine samples was 10.8% (95% CI: 7.2% - 15.9%) , which was significantly higher than that in the swab samples (7.8%, 95% CI: 4.9% - 12.1%; χ2 = 39.2, P < 0.05) . Compared with the swab sample-based CT-DNA detection method, the sensitivity, specificity, positive predictive value, negative predictive value and consistency rate of the urine sample-based CT-DNA detection method were 97.0% (128/132) , 96.3% (1 293/1 343) , 71.9% (128/178) , 99.7% (1 293/1 297) , and 96.3% (1 421/1 475) , respectively. The positive rate of CT DNA in the urine samples from 1 118 male patients was 11.0% (95% CI: 7.2% - 16.5%) , which was significantly higher than that in the swab samples (7.6%, 95% CI: 4.9% - 11.8%; χ2 = 34.3, P < 0.05) . There was no significant difference in the positive rate of CT DNA between the urine (11.9%, 95% CI: 7.7% - 17.9%) and cervical swab samples from 357 female patients (10.4%, 95% CI: 7.6% - 14.0%; χ2 = 3.2, P > 0.05) . Conclusions:The positive rate of CT DNA in urine samples is higher than or similar to that in urethral/cervical swab samples. The urine sample-based CT-DNA detection method has characteristics of convenience, non-invasiveness, painlessness and low cost, and is worthy of clinical promotion.