Objective To discuss the preoperative and postoperative holistic care for patients with ectopic pregnancy. Methods A retrospective study was carried out in preoperative and postoperative holistic care and detailed discharge instruction of 64 patients with ectopic pregnancy. Results In addi-tion to 2 cases of conservative treatment, the remaining 62 cases received satisfactory treatment, the hospi-talization time lasted 7 to 14 days. Conclusions Skilled technique, sturdy nursing knowledge and proper psychological nursing of nurses can benefit early rehabilitation of patients, and preoperative and postopera-tive psychological nursing is the key of rehabilitation.

Objective To study the expression of malignant melanoma(MM) related genes by cDNA microarray technique. Methods mRNA, extracted from tissues of patients and normal controls, was reversely transcripted into cDNA and marked with 33P. The cDNA probes were hybridized to cDNA microarrays, which contained 2000 human genes each array. The down-regulation of two co-differentiated expressed genes was confirmed by quantitative real-time RT-PCR. Results Different expression between MM and normal controls was found in 4.7%-6.15% of genes by more than 2 times,0.75%-1.4% by more than 5 times, and 0.45-0.5% by more than 10 times. These genes were pro-oncogenes, tumor suppressor genes, genes related to apoptosis, cell cycle related genes, and so on. Three genes were down-regulated in all of the patients. Two of those genes, histidine triad nucleotide-binding protein (HINT) and RBP1-like protein (BCAA), were down-regulated, as identification by quantitative real-time RT-PCR. Conclusions cDNA microarray can be used effectively to reveal melanoma gene expression profiling for the propose of carcinogenesis study. HINT and BCAA are the first reported genes down-regulated in MM. However, further studies are needed for their expressive specificity and mechanism in MM.

Objective To investigate the role of mutated BRAFV599E gene in the growth of malignant melanoma cells. Methods In the previous study, plasmids containing small hairpin RNAs (shRNAs), braf1 and braf2 specific for mutated BRAFV599E gene, were designed and used to transfect A375 cells to inhibit the expression of BRAF gene in these cells. In this study, four kinds of A375 cells, including Abraf1 (transfect ed with braf1), Abraf2 (transfected with braf2), Aneg (transfected with negative plasmid) and A375 (untransfected) cells, were chosen and cultured in 96-well plate. MTT assay, plate clone forming assay, flow cytometry were applied to test the growth, clone formation, cell cycle and apoptosis of these cells respectively. Results Compared with A375 and Aneg cells, inhibited proliferation (F=25.48, P<0.001) and clone-forming rate (F=90.06, P<0.001) were observed in Abraf1 and Abraf2 cells; furthermore, flow cytometry showed a decrease in S-phase population(F=147.87, P<0.001) but an increase in G1-phase population (F=9.14, P<0.05)in Abraf1 and Abraf2 cells. However, neither Abrafl nor Abraf2 cells exhibited a significant increase in apoptosis ratio (F=2.27, P>0.05). Conclusions Mutated BRAFV599E gene could induce the switch from G1 phase to S phase in melanoma cells, subsequently accelerate the growth of melanoma cells, but it has no obvious influence on the apoptosis of these cells.

With anti-myc and ras oncogene product monoclonal antibodies p62 and p21, 27 cases of squamous epithelial tumors of the skin (SETS) specimens were examined. The results showed that 62.9%(17/27) and 66.7%(18/27) of the specimens expressed p62 and p21 proteins respectively. Thirteen specimens expressed p62 and p21 proteins simultaneously. The results also showed that p62 was mainly seen in the poorly differentiated squamous cell carcinoma specimens, while p21 was mainly in the relatively well-differentiated specimens. The authors consider that myc and ras oncogenes might have different effects on the develpment of the SETS, and their synergy might be associated with the persistant growth of SETS.

Objective To investigate the involvement of apoptosis in the lesions of patients with psoriasis. Methods The apoptosis was detected with terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL), and the expression of p53, PCNA and apoptosis suppressing protein bcl 2 was assessed with immunoperoxidase technique in psoriatic lesions and normal skin. Results A large number of keratinocytes showing biochemical and morphologic features of cells undergoing apoptosis were observed in all the suprabasal layers of the psoriatic epidermis. The plaques from all patients analysed showed marked increase in the number of PCNA positive cells in the middle and basal keratinocytes, and a dramatic reduction in the number of bcl 2 positive cells in the basal cell layer. Conclusion The increased apoptosis of keratinocytes in the lesions of psoriasis might be a homeostatic mechanism to the hyperplasia of cells.

Objective To assess the clinical feature and diagnosis of diffuse hyperpigmentation with guttate depigmentation macules.Methods A retrospective study was carried out among 10 patients with diffuse hyperpigmentation with guttate depigmentation macules collected at the Institute of Dermatology,Chinese Academy of Medical Sciences and Peking Union Medical College from 2005 to 2012.The clinical manifestations,pathological findings and disease outcomes in these patients were analyzed.Results Of the 10 patients,6 were male,and 4 were female.The median age at onset was 7 years (range,4-25),and there were only 3 adult patients among these patients.None of the patients had a family history of pigmentary disturbance.Typical clinical manifestations included densely distributed,guttate hypopigmented macules arising on diffuse and uniform hyperpigmentation.Lesions could slowly spread over the body surface without the trend towards spontaneous regression.Pathologically,there was a slight increase in pigmentation of the epidermal basal layer,as well as melanins and melanophages scattered around blood vessels in the superficial dermis,with or without focal vacuolar degeneration of the basal cell layer.Conclusions Diffuse hyperpigmentation with guttate depigmentation macules,a rare pigmentary disturbance that clinically manifests as both hyperpigmentation and hypopigmentation and is pathologically characterized by postinflammatory hyperpigmentation,often affects children.Once the lesions occur,there is no trend towards regression.No effective treatment is available for this entity at present.

Objective To investigate the clinicopathologic features and differential diagnosis ofacroangiodermatitis (AM).Methods Clinical and pathological data on 12 patients with AM were retrospectively reviewed.Results Clinical manifestations of AM consisted of circumscribed brown to violaeeous macules,plaques,nodules and ulceration.Lesions were located in bilateral legs in 6 patients,and in unilateral legs in the other 6 patients.Histopathological examination revealed an increased number of lobular or clump-shaped capillaries and small veins whose lumens were round and regular,swelling of vascular endothelial cells,and different degrees of erythrocyte extravasation,hemosiderin deposition,dermal fibrosis and sparse infiltrates of inflammatory cells.The lesions were histologically located in the superficial dermis in 3 cases,in the upper and middle dermis in 8 cases,and in the entire dermis in 1 case.Immunohistochemical studies showed that vascular endothelial cells stained positive for CD31 and CD34,while perivascular cells stained negative for CD34.Conclusions AM has specific clinical and pathological manifestations,and pathological examination is essential for the diagnosis of AM.

Objective To study the inhibitory effects of siRNA targeting survivin on the expression of survivin,as well as the apoptosis,proliferation and invasion of a human melanoma cell line,M14.Methods Two siRNAs targeting survivin were designed,chemically synthesized,and used to construct the recombinant plasmids,pRAT-H1.1/neo-survivin-siRNA1 and pRNAT-H1.1/neo-survivin-siRNA2.Then,recombinant plasmids were transfected into M14 cells mediated by Lipofectamine 2000 reagent.Those cells untransfected or transfected with empty vector served as the control.After culture over various periods of time.cells were collected for the detection of mRNA and protein expression of survivin with RT-PCR and Western blotting,respectively,and for the examination of apoptosis and proliferation of M14 cells by flow cytometry and MTT methods,respectively.Also,Transwell assay was performed to detect the invasive capability of M14 cells.Results A statistical decrease in the mRNA and protein expressions of survivin was observed along with an increase in apoptotic rate(x2=31.55,P＜0.01)in M14 cells transfectcd with siRNA-containing plasmid compared with untransfected and empty vector-transfected cells.As MTT assay indicated,on day 4 after the transfcorion,the proliferation of M14 cells was inhibited by(55.4±4.3)%,(34.5±4.3)%and(13.3±4.6)%,with pRNAT-H1.1/neo-survivin-siRNA1,pRNAT-H1.1/neo-survivin-siRNA2 and empty vector,respectively:there was a significant difference among the three groups(P＜0.05).Decreased invasive capability was noticed in M14 cells transfected with siRNA-containing plasmid compared with untransfected cells(all P＜0.05).Conclusions The plasmid containing siRNA against survivin can specifically inhibit the expression of sarvivin,proliferation and invasion of tumor cells,and induce cell apoptosis.The inhibition of survivin expression by siRNA may be a rational approach to the gene therapy for malignant melanoma.

Objective To construct the pCDNA3.1-ING4 eukaryotic expression vector and investigate its effect on the proliferation of melanoma cell line M14. Methods The targeted cDNA fragment encoding ING4 was cloned by reverse transcription-PCR with normal gastric mucosa from patients with gastric ulcer, and subcloned into eukaryotie expression vector pcDNA3.1. PCR and DNA sequencing were performed to identify the eukaryotic expression vector pCDNA3.1-ING4, which was then transfected into M14 cells with Lipofectamine 2000 reagent. The expression of ING4 was detected in untransfected, pCDNA3. 1-ING4-transfected and pcDNA3.1-transfected M14 cells by Western blot and immunocyto-chemistry, and cell proliferation by MTT assay. Results A fragment of expected size (750 bp) was amplified by PCR analysis, and DNA sequencing confirmed the correctness of the recombinant plasmid. As shown by immunocytochemistry, the percentage of cells positive for ING4 protein was significantly higher in pCD NA3. 1-ING4-transfected MI4 cells than in non-transfected M14 cells and pcDNA3.1-transfected M14 cells (71.80%±9.88% vs 4.20%±3.35%, P < 0.01). Western blot also revealed an increased expression of ING4 in pCDNA3.1-ING4-transfected cells. Decreased cell viability was observed in ING4-transfected cells compared with nontransfected cells and pCDNA3.1-transfected cells (both P < 0.01). Conclusions The pCDNA3. 1-ING4 eukaryotic expression vector has been constructed successfully, and M14 cells transfected by this recombinant plasmid could effectively express ING4 protein, which may inhibit the cell proliferation of M14 cells.

The clinical course of mycosis fungoides is indolent except when large cell transformation occurs. Large cell transformation of MF is rare and easy to misdiagnose. A case of large cell transformation of mycosis fungoides is reported. A 40-year-old man presented with a 10-year history of pruritic erythema and papules in the trunk and extremities as well as a 5-month history of nodules on the nape of the neck.Histopathologically, the erythematous patch showed typical changes of mycosis fungoides, while the tumor cells were small and expressed CD3 and CD4, and only a small number of tumor cells expressed CD30. Pathological examination of nodular lesions revealed the infiltration of large pleomorphic lymphoid cells expressing CD3 and CD4 throughout the entire dermis. There was an epidermotropism of large cells, and about 40% of these cells expressed CD30. Based on the medical history and histological findings, the patient was diagnosed with large cell transformation of mycosis fungoides. The lesions improved markedly after 3-week treatment with oral acitretin (30 mg once daily), subcutaneous interferon-alpha (2 × 106 IU thrice a week) and local superficial X-ray irradiation for nodular lesions. Up to the time of this writing, the patient had been followed.