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.

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 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.

Objective To investigate the effects of NB-UVB on the expression of Gadd45α as well as cell apoptosis and cycle of human HaCaT keratinocytes.Methods Cultured HaCaT cells were exposed to various doses (100,200,400 mJ/cm2)of NB-UVB followed by an additional culture of 6,12 and 24 hours,respectively.Reverse transcription PCR and Western blot were performed to detect the mRNA and protein expression of Gadd45α respectively in HaCaT cells,cell counting kit 8 (CCK8)to measure the proliferation of cells,and flow cytometry to determine the cell cycle distribution of HaCaT cells before and after the exposure to NB-UVB.Results Gadd45α was expressed in HaCaT cells.After exposure to NB-UVB of the three doses,the mRNA and protein levels of Gadd45α increased at 6 hours and 12 hours,but declined at 24 hours,and significant changes were observed in HaCaT cells at the three time points after exposure to NB-UVB of the three doses (all P<0.05).The Gadd45α/β-actin mRNA ratio was 1.4360±0.6551.1.8633±0.0979,1.9266±0.1724 in HaCaT cells 12 hours after irradiation to NB-UVB of 100,200 and 400 mJ/cm2,respectively,significantly higher than that in unirradiated cells(0.6000±0.1276,all P<0.05).Also,increased Gadd45α/β-actin protein ratio was noted in HaCaT cells 12 hours after irradiation to NB-UVB of 100,200 and 400 mJ/cm2 compared with unirradiated cells (0.0773±0.0005,0.1936±0.0015,0.2373±0.0015 vs.0.0290±0.0010,all P<0.05).NB-UVB inhibited the proliferation of HaCaT cells in a time-and dose-dependent manner.Flow cytometry showed that irradiated HaCaT cells were blocked in G2 phase of the cell cycle.and the percentage of HaCaT cells in G2 phase was 13.53%±1.03%,17.77%±2.25%,30.03%±4.29%afler exposure to NB-UVB of 100,200 and 400 mJ/cm2,respectively,compared to 9.24%±0.97%in unirradiated cells (all P<0.05).Conclusions The expression of Gadd45α is increased in HaCaT cells after exposure to NB-UVB,and Gadd45α may be involved in the NB-UVB-induced suprression of cell proliferation of and cell cycle arrest in HaCaT cells.

Objective To identify the gene locus and the mutation of DSRAD (double-stranded RNA adenosine deaminase) in a Chinese dyschromatosis symmetrica hereditaria(DSH) family. Methods After confirming the diagnosis of the DSH proband, the genomic DNA was extracted from the whole blood samples of every members of the pedigree. The DSRAD gene intervals were localized by linkage analysis and haplotype reconstruction. The mutation of DSRAD was detected by direct sequencing. Results The candidate gene was localized at the 1q region, consistent with the reported region. The direct sequencing results showed that there was a CAA→TAA transition at exon 2 of DSRAD in all affected family members, which consequently led to a nonsense mutation of Gln517Ter. Conclusion A nonsense mutation is found in the Chinese DSH family.

Objectives To synthesize human telomerase reverse transcriptase (hTERT)- and human tolemerase RNA (hTR)- small interfering RNA (siRNA) and investigate their effects on telomerase activity in the cutaneous T-cell lymphoma (CTCL) cell line Hut78. Methods Two types of hTERT- and hTR- siRNAs were synthesized with T7 RNA polymerase via in vitro transcription, then either mixed with Hut78 cell lysates directly or transfected into Hut78 cells by calcium phosphate co-precipitation. Telomerase activity was tested by telomeric repeat amplification and polyacrylamide gel electrophoresis. Results With T7 RNA polymerase, hTERT- and hTR- siRNAs were synthesized efficiently with a concentration of 22.4?g siRNA per 40?L siRNA reaction mix. Telomerase activity was suppressed significantly by either of the siRNAs. The inhibition rate was 87% in the cell lysate group treated with siRNA directly, and 75% in the cell group Iransfected with siRNA. Conclusions The in vitro transcription of siRNA with T7 RNA polymerase is technically simple, costeffective, and can produce siRNA in an efficient way. hTERT- and hTR-siRNA can down-regulate telomerase activity significantly in Hut78 cells.