Detection of Chlamydia trachomatis DNA in urine and urethral/cervical swab samples: analysis of results from 1 475 outpatients attending sexually transmitted disease clinics / 中华皮肤科杂志

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.

Effect of nucleolar protein 14 on angiogenesis in melanoma and its mechanism / 中华皮肤科杂志

Objective To evaluate the effect of nucleolar protein 14 (NOP14) on angiogenesis in melanoma.Methods Melanoma tissues were collected from 40 patients with pathologically diagnosed melanoma in Guangzhou First People's Hospital from January 2016 to December 2018,and immunohistochemical study was conducted to determine the expression of NOP14 and CD31 (expressed as microvessel density [MVD]).Melanoma cell lines A375 and SK-MEL-1 were both divided into 4 groups:empty vector group transfected with the empty vector,NOPI4 group transfected with a NOP14-overexpressing vector,siNOP14 group transfected with the siRNA targeting NOP14,and siNC group transfected with a negative control siRNA.Fluorescence-based quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of NOP14 respectively,and Western blot analysis and enzyme-linked immunosorbent assay (ELISA) to measure the expression of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) in cells and their culture media.Coculture models of human umbilical vein endothelial cells (HUVECs) and A375/SK-MEL-1 cells in the above groups were established in Transwell chambers,and cell counting kit-8 (CCK8) assay,Transwell migration and invasion assays and Matrigel-based vasculogenic mimicry assay were performed to evaluate the cellular proliferative,migratory,invasive activity and tube formation capacity respectively.A linear regression model was used to analyze the relationship between NOP14 expression and MVD in melanoma tissues,multi-way analysis of variance to analyze the difference in cellular proliferative activity,and independent-sample t test to compare other experimental indices between 2 groups.Results The expression of CD31 (MVD) was 44 ± 13 in the group with high NOP14 expression (n =20),58 ± 16 in that with moderate NOP14 expression (n =17),and 62 ± 11 in that with low NOP14 expression (n =3).The NOP14 expression was negatively correlated with MVD (r =-0.525,P =0.017).Compared with the empty vector group,the expression of VEGF and VEGFR in A375 and SK-MEL-1 cells and their culture media significantly decreased in the NOP14 group (all P ＜ 0.05).Compared with the siNC group,the expression of VEGF and VEGFR in the A375 and SK-MEL-1 cells and their culture media significantly increased in the siNOP14 group(all P ＜ 0.05).In the co-culture models of A375 cells and HUVECs,the NOP14 group showed significantly decreased proliferative activity of HUVECs (F =131.85,P ＜ 0.05),and numbers of migratory cells (22 ± 5 vs.63 ± 8,t =7.07,P =0.002),invasive cells (14 ± 5 vs.45 ± 10,t =4.94,P =0.008) and branch points (8 ± 2 vs.14 ± 3,t =5.06,P ＜ 0.001) compared with the empty vector group;compared with the siNC group,the siNOP14 group showed significantly increased proliferative activity of HUVECs (F =79.92,P ＜ 0.01),and numbers of migratory cells (152 ± 30 vs.59 ± 4,t =5.36,P =0.006),invasive cells (134 ± 21 vs.50 ± 8,t =6.40,P ＜ 0.001) and branch points (27 ± 3 vs.15 ± 4,t =6.10,P ＜ 0.001).In the co-culture models of SK-MEL-1 cells and HUVECs,the 4 groups showed the same trend of changes in the cellular proliferative,migratory,invasive activity and tube formation capacity of HUVECs as the above groups in the co-culture models of A375 cells and HUVECs.Conclusion The NOP14 expression is negatively correlated with MVD in melanoma tissues,and NOP14 can inhibit angiogenesis in melanoma.

miR-122-5p inhibits the proliferation of melanoma cells by targeting NOP14 / 南方医科大学学报

OBJECTIVE@#To investigate the expression profile of miR-122-5p in melanoma tissues and the effect of miR-122-5p on the proliferation, cell cycle and apoptosis of human melanoma cell lines SK-MEL-110 and A375.@*METHODS@#The expression profiles of miR-122-5p in melanoma and pigmented nevus tissues were detected using real-time fluorescence quantitative PCR (qRT-PCR). SK-MEL-110 and A375 cells transfected with miR-122-5p inhibitor or negative control inhibitor (NC) I were examined for miR-122- 5p expression using qRT-PCR and changes in cell proliferation, cell cycle and apoptosis using MTT assay or flow cytometry. NOP14 mRNA and protein expressions in the cells were detected using qRT- PCR and Western blotting, respectively. Luciferase reporter assay was used to confirm the identity of NOP14 as the direct target of miR-122-5p.@*RESULTS@#The relative expression of miR-122-5p in human pigmented nevus tissues and melanoma tissues was 1.23±0.270 and 7.65 ± 1.37, respectively. The relative expression of miR-122-5p in SK-MEL-110 and A375 cells transfected with miR-122-5p inhibitor was 0.21 ± 0.08 and 0.17 ± 0.05, respectively. miR-122-5p inhibitor obviously inhibited the cell proliferation and increased the percentage of cells in G1 stage in both SK-MEL-110 and A-375 cells, but did not cause obvious changes in the apoptosis of the two cells. miR-122-5p inhibitor did not significantly affect the expression level of NOP14 mRNA, but obviously increased the expression level of NOP14 protein. Luciferase reporter assay revealed a significantly lower luciferase activity in cells co-transfected with miR-122-5p mimics and wild-type psi-CHECK2-3'UTR plasmid than in the cells cotransfected with NC and wild-type psi-CHECK2-3'UTR plasmid (0.21 ± 0.14 0.56 ± 0.1, < 0.01).@*CONCLUSIONS@#miR-122-5p expression is upregulated in melanoma tissues, indicating its involvement in the development of melanoma. miR-122-5p inhibits the proliferation of SK-MEL-110 and A-375 cells possibly by affecting the cycle through NOP14.

Curative Effect of Micro Plasma on Facial Acne Atrophic Scar / 现代医院

Objective To observe the clinical efficacy of micro plasma in the treatment of acne atrophic scar on face.Methods 82 cases with facial acne atrophic scar were randomly divided into the observation group and the control group.Each group had 41 cases.The observation group received micro plasma treatment while the control group were treated with ultrapulsed fractional CO2.The treatment was carried out 6 weeks interval, in total 3 times.After each treatment, the curative effect and the side effects were compared between the two groups.Results The total effective rates had no statistical difference between two groups, and the more treatment times, the better effect.The observation group had less adverse effects, which the duration of main side effects was shorter in the observation group than in the control group, with statistically significant difference.Conclusion The micro plasma technology on treating facial acne atrophic scar was an effective skill with less adverse reactions and worth of wide use.

Expression of Caspase 8 and phospho-Akt in condyloma acuminatum lesions / 中华皮肤科杂志

Objective To determine the expression of Caspase 8 and phospho-Akt(p-Akt)in condyloma acuminatum(CA)lesions, and to evaluate their significance. Methods Skin lesion samples were collected from 30 patients with CA, cancer tissue samples from 20 with cervical cancer, and normal skin samples from 20 healthy controls. All the samples were subjected to paraffin embedding. An immunohistochemical study was conducted to determine the expression and distribution of Caspase 8 and p-Akt in the above samples. Results The expression rate of Caspase 8 was significantly lower in CA lesions (23.33%)than in normal skin samples(90%, P < 0.01)and cervical cancer lesions(80%, P < 0.001). Moreover, the expression rate of p-Akt in CA lesions(93.33%)was significantly higher than that in the normal skin samples(90%, P<0.001), but lower than that in the cervical cancer lesions(95%, P<0.001). No significant correlations were observed between the expression of Caspase 8 and p-Akt in either CA lesions or normal skin samples. However, the expression of Caspase 8 was positively correlated with the expression of p-Akt in cervical cancer lesions(r=0.369, P<0.05). Conclusion Both suppressed apoptosis initiation of Caspase 8 and anti-apoptotic effect of p-Akt may be involved in the occurrence and development of CA.

Effects of miR-145 on the proliferation, apoptosis and cell cycle of a human keratinocyte cell line HaCaT / 中华皮肤科杂志

Objective To investigate the regulatory effects of miR-145 on the proliferation,cell cycle and apoptosis of a human keratinocyte cell line HaCaT.Methods miR-145 mimics and negative control (NC) mimics were chemically synthesized and then transiently transfected into HaCaT cells respectively.After additional culture for different durations,real-time PCR was performed to determine the expression level of miR-145,MTS assay to estimate cell proliferation,and flow cytometry to detect cell apoptosis and cycle.Luciferase assay,real-time PCR and Western blot were conducted to determine whether NRAS was the target gene of miR-145.Results The miR-145 expression level in miR-145 mimic-transfected cells increased by 85.00 ± 1.21 folds compared with NC mimic-transfected cells (t =115.90,P ＜ 0.0001).The transfection with miR-145 mimics significantly inhibited the proliferation of HaCaT cells (F =8.76,P =0.008),and the inhibitory effect significantly varied with the duration (24-96 hours) of culture after transfection,with no interaction effect between the transfection with miR-145 mimics and culture duation (F =1.21,P =0.18).Compared with NC mimic-transfected cells,those transfected with miR-145 mimics showed a significant increase in the proportion of early apoptotic cells (18.9％ ± 4.1％ vs.4.3％ ± 1.2％,t =7.126,P ＜ 0.01),late apoptotic cells (9.3％ ± 2.3％ vs.3.6％ ± 1.6％,t =12.38,P ＜ 0.01),G1-phase cells (85.83％ ± 5.2％ vs.62.08％ ± 6.23％,t =11.78,P =0.007),but a significant decrease in the percentage of G2-phase cells (6.26％ ± 1.2％ vs.19.36％ ± 3.45％,t =7.610,P =0.017) and S-phase cells (7.91％ ± 1.3％ vs.18.56％ ± 5.23％,t =7.230,P=0.019).As luciferase assay showed,luciferase activity was significantly lower in HaCaT cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-wild carrying the wild-type 3'UTR of NRAS than in those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-wild (t =11.09,P =0.008),but similar between cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-mut carrying the mutant-type 3'UTR of NRAS and those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-mut (P ＞ 0.05).Real-time PCR and Western blot revealed that the overexpression of miR-145 mimics had no significant effect on NRAS mRNA expression (P ＞ 0.05),but significantly inhibited NRAS protein expression (1.52 ± 0.07 vs.0.20 ± 0.02,t =28.43,P＜ 0.01).Conclusion miR-145 might inhibit proliferation and promote apoptosis of HaCaT cells by influencing cell cycle via NRAS.

Expression of Toll-like receptor 4 (TLR4), TLR9 and DC-specific intracellular adhesion molecule-3 grabbing non-integrin (SIGN) in condyloma acuminatum lesions / 中华皮肤科杂志

Objective To investigate the expression and significance of TLR4, TLR9 and DC-SIGN in primary and recurrent condyloma acuminatum (CA) lesions. Methods An immunohistochemical method using streptavidin-peroxidase (SP) was performed to detect the expressions and distribution of TLR4, TLR9 and DC-SIGN in tissue specimens obtained from the recurrent CA lesions of 30 patients, primary CA lesions of 30 patients, and from the foreskin of 20 normal human controls. Results The expression levels of TLR4, TLR9 and DC-SIGN in primary and recurrent CA lesions were significantly higher than those in normal control tissue (all P < 0.001), and the cells expressing TLR4, TLR9 or DC-SIGN were mainly located in the basal and spinous layer in CA lesions. There was no significant difference in the expressions of TLR4, TLR9 or DC-SIGN between primary and recurrent CA lesions (all P> 0.05). A positive correlation was found between the expression of TLR4, TLR9 and DC-SIGN in CA lesions. Conclusion The overexpression of TLR4, TLR9 and DC-SIGN probably plays an important role in the occurrence and recurrence of CA.

Treatment of 60 Cases of Senile Herpes Zoster by Encircled Acupuncture plus Valaciclovir / 针灸推拿医学（英文版）

To observe the clinical therapeutic effect and safety of local encircled acupuncture plus valaciclovir in treating senile herpes zoster.Methods:Sixty senile patients with herpes zoster were divided into two groups.In acupuncture and medicine group,the patients were treated by encircled acupuncture plus valaciclovir.The needle was inserted about 0.8 cun away from herpes and to form an angle of 15°with the skin around the skin lesion.During treatment,valaciclovir was taken orally 300 mg every time,twice every day for successive 10 days.In westem medicine group,valaciclovir was taken orally 300 mg every time,twice every day for successive 10 days.Results:The time of stopping herpes,relieving pain andscabbing in acupuncture and medicine group was significantly lower than that in western group.Conclusion:Local encircled acupuncture plus valaciclovir in treating senile herpes zoster got effects quickly and could effectively shorten the course of disease and reduce the incidence rateofresidual neuralgia.