Objective To investigate the effect of targeted interruption of cyclooxygenase-2 (COX-2) gene by small interference RNA (siRNA) on the expression of COX-2, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in eutopic and ectopic endometrial stromal cells (ESC) with endometriosis, and the effect on the content of 6-keto-prostaglandin-F1α (6-keto-PGF1α, metabolites of COX) and the apoptosis of eutopic and ectopic ESC with endometriosis. Methods Ectopic and eutopic ESC from 30 women with endometriosis were isolated and cultured respectively. Then, ESC were classified into three groups: interference group, negative control group and blank control group. ESC in interference group were injected into siRNA transfection complex while ESC in negative control group were injected into negative control transfection complex. ESC from 10 participants without endometriosis were the normal control group. The mRNA and protein expression of COX-2, VEGF, MMP-9 in pre-transfected and post-transfected eutopic and ectopic ESC were detected through real time reverse transcription PCR and western blot. The content of 6-keto-PGF1α was determined by ELISA, the apoptotic cells were detected by flow cytometry. Results After interruption of COX-2 gene, there were no significant difference in the mRNA and protein expression of COX-2, VEGF and MMP-9 between the negative control group and blank control group (P>0.05); the mRNA and protein expression of the three genes in interference group were significantly lower than those in negative control group and blank control group (P<0.05); the mRNA expression of the three genes in interference group of eutopic ESC were 0.87±0.06, 1.76±0.59, 1.04±0.32, in interference group of ectopic ESC were 0.75±0.12, 1.62±0.47, 0.88±0.25, the protein expression of the three genes in interference group of eutopic ESC were 0.457 ± 0.019, 0.500 ± 0.012, 0.361 ± 0.008, in interference group of ectopic ESC were 0.323 ± 0.018, 0.474 ± 0.016, 0.339 ± 0.009;the mRNA and protein expression of the three genes in ectopic ESC had a more reduction than those in eutopic ESC (P<0.05). The results from ELISA revealed that the content of 6-keto-PGF1α in the normal control group [(17.7 ± 1.9) pg/ml] were significantly lower than those in the blank control group (P<0.05), the content of 6-keto-PGF1α in ectopic ESC were significantly higher than that in eutopic ESC (P<0.05), the content of 6-keto-PGF1α in the blank control group of eutopic and ectopic ESC were (32.4±2.6) pg/ml, (38.2±3.7) pg/ml;there was no significant difference in the content of 6-keto-PGF1α between the negative control group and blank control group (P>0.05);compared with those of negative control group and blank control group, the content of 6-keto-PGF1αin interference group decreased significantly (P<0.05), the content of 6-keto-PGF1α in interference group of eutopic and ectopic ESC were (17.1 ± 2.4) pg/ml, (20.9 ± 2.7) pg/ml; the content of 6-keto-PGF1α in eutopic ESC had a slightly more reduction than that in ectopic ESC (P>0.05). The results from flow cytometry displayed that, there was no significant difference in apoptotic cells between the negative control group and blank control group (P>0.05);compared with those of negative control group and blank control group, more apoptotic cells were detected in interference group and the difference was significant (P<0.01);the apoptotic cells in ectopic ESC were significantly more than that in eutopic ESC (P<0.05); the apoptosis rate in interference group of eutopic and ectopic ESC were (33.76 ± 0.06)%, (47.18 ± 0.12)%. Conclusions Our results suggested the targeted interruption of COX-2 gene by siRNA effectively inhibited the mRNA and protein expression of COX-2, VEGF and MMP-9 in both eutopic ESC and ectopic ESC with endometriosis, greatly increased the apoptotic rate of cells and obviously reduced the content of 6-keto-PGF1αby inhibiting the activity of COX-2. And the changes in ectopic endometrium were more evident than those in eutopic endometrium.

Objective:To analyze clinical and histopathological characteristics of infantile congenital melanocytic nevi (ICMN) .Methods:Clinical and pathological data were collected from 126 infants with confirmedly diagnosed congenital melanocytic nevi in Department of Dermatology, Xijing Hospital from January 2015 to January 2020, and were retrospectively analyzed. Chi-square test was used for comparisons of enumeration data.Results:Among the 126 patients with ICMN, 68 were males and 58 were females; 109 (86.5%) presented with skin lesions at birth; 73 (57.9%) were 2 - 3 years old at the first clinic visit. The skin lesions occurred on the head and face (76 cases, 60.3%) , trunk (24 cases, 19.1%) or extremities (26 cases, 20.6%) . There were 36 (28.6%) patients with small congenital nevi, 68 (54.0%) with M1-type medium-sized nevi, 13 (10.3%) with M2-type medium-sized nevi and 9 (7.1%) with giant nevi. Of 126 cases of ICMN, 121 (96.0%) had solitary lesions, 5 (4.0%) had multiple lesions, 44 (34.9%) had nevi with coarse hairs, 15 (11.9%) had nevi complicated by papules or hyperplastic nodules, and 6 (4.8%) had satellite lesions. Pathological subtypes included compound nevus (120 cases, 95.2%) , intradermal nevus (4 cases, 3.2%) , and junctional nevus (2 cases, 1.6%) . Under the microscope, the depth of the skin lesions was < 1 mm in 38 (30.1%) cases, 1 - 2 mm in 61 (48.4%) and > 2 mm in 25 (19.8%) , and 45 (35.7%) cases showed nevus cells infiltrating the subcutaneous fat layer or deeper tissues. Among the 126 ICMN lesions, common pathological features included nevus tissue maturation (100%, 2 cases of junctional nevi were excluded) , pigment granules in the stratum corneum (53 cases, 42.1%) , disordered/asymmetric distribution of nevus cells (80 cases, 63.5%) , scattered epidermal nevus cells (91 cases, 72.2%) , pagetoid spread of epidermal nevus cells (67 cases, 53.2%) , melanophages in the dermis (71 cases, 56.4%) , and nevus cells distributed along hair follicles/sebaceous glands (82 cases, 65.1%) . Special pathological features included nevus cells embedded in the vascular/lymphatic vessels (42 cases, 33.3%) , nevus cell lysis (45 cases, 35.7%) , fibromatous changes (25 cases, 19.8%) , involvement of the arrector pilli muscles (31 cases, 24.6%) , and mast cell infiltration (30 cases, 23.8%) . Pathological patterns of ICMN with different clinical features: the incidences of infiltration depth > 2 mm, pigment granules and columnar pigment granules in the stratum corneum were significantly higher in the giant nevi than in the small and medium-sized nevi ( χ2 = 7.93, 10.76, 5.89 respectively, all P < 0.05) ; the incidences of infiltration depth > 2 mm, epidermal spongiosis with scattered nevus cells, nevus cell nests distributed along the hair follicles/sebaceous glands, fibromatous changes and mast cell infiltration were significantly higher in the skin lesions with coarse hairs than in those without ( χ2 = 28.29, 8.11, 6.22, 7.92, 8.19 respectively, all P < 0.01) ; the incidences of pagetoid spread of epidermal nevus cells and atypical nevus cells were significantly higher in the skin lesions with papules/hyperplastic nodules than in those without papules/hyperplastic nodules ( χ2 = 4.92, 6.30 respectively, both P < 0.05) . Conclusions:The clinical and histopathological characteristics of ICMN are unique, and atypical nevus cells are common in ICMN. The diagnosis and treatment of ICMN need to be based on the combination of clinical and pathological characteristics.