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 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 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 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.

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 estimate the diagnostic value of interstitial infiltration pattern for early morphea.Methods Twenty-five cases of early morphea pathologically characterized by interstitial infiltration of inflammatory cells were collected from 2010 to 2012.The clinicopathological features of these cases were retrospectively analyzed.Results The average clinical course was 7.5 months.The primary manifestation was edematous dark erythematous plaques,and interstitial or mixed infiltrate of inflammatory cells was the characteristic histopathological presentation.After anti-inflammatory treatment,lesions markedly improved or disappeared in 70％ of these patients.Conclusions Interstitial infiltration of inflammatory cells is a rare histologic pattern in early morphea.To learn and recognize this pattern may be beneficial to the diagnosis and treatment of early morphea.

Objective To study the effects of triptolide on the apoptosis in and proliferation of a human melanoma cell line M14.Methods M14 cells were cultured with the presence of 5 concentrations (12.5,25,50,100,200 nmol/L) of triptolide for 24,48 and 72 hours respectively,and cell counting kit-8 (CCK-8) was used for the detection of cell proliferation.Some M14 cells were treated with triptolide at 10 nmol/L,20 nmol/L and 30 nmol/L for 48 hours followed by the analysis of cell cycle by flow cytometry and detection of cell apoptosis by flow cytometry following annexin V-fluorescein isothiocyanate (FITC)/propidium iodide double staining.The morphological changes of M14 cells treated by triptolide at 30 nmol/L for 48 hours were observed by Hoechest 33258 staining.Results Compared with untreated M14 cells,an increase of cell population in S phase was observed in triptolide-treated cells,along with a decline in cell population in G2/M phase.The apoptosis rate was (2.92 ± 0.17)％,(20.99 ± 0.40)％,(34.28 ± 2.04)％ and (63.38 ± 0.71) ％ respectively in M14 cells treated with triptolide at 0,10,20 and 30 nmol/L for 48 hours,suggesting that triptolide enhanced the proliferation of M14 cells in a dose-dependent manner.After treatment with triptolide of 30 nmol/L,M14 cells showed morphological changes characteristic of apoptosis.Conclusion Triptolide could inhibit the proliferation of and induce the apoptosis in M14 human melanoma cells.

Objective To compare three methods for the extraction of mycobacterial DNA.Methods Two commercial DNA extraction kits and an ordinary freeze-thawing method were used to extract DNA from the pure suspensions of three species of Mycobacteria (M.tuberculosis,M.leprae and M.smegmatis) at different densities (1 × 10 to 1 × 105 cells/ml),simulated clinical specimens containing different concentrations of mycobacterial cells (1 × 10 to 1 × 104 cells/ml).The purity and concentration of the extracted DNA were evaluated.Then,PCR was performed to amplify the 16S rRNA region of Mycobacteria.The performance of the three methods was compared by the purity and concentration of extracted DNA as well as the results of PCR.Further more,76 clinical skin specimens suspected to be infected with Mycobacteria were used to further validate the performance of these methods.Results All the extracted DNA samples could be detected by PCR.The highest purity of DNA was obtained by the kit A,followed sequentially by the freeze-thawing method and the kit B.When pure suspensions were used,the detection limit was consistently 1 × 102 cells/ml for all the three methods.With simulated specimens,the detection rate was consistently 100％ for all the three methods at the concentration of 1 × 103 cells/ml,60％ (12/20),55％ (11/20) and 55％ (11/20) for the kit A,kit B and freeze-thawing method respectively at the concentration of 1 × 102 cells/ml.The analysis of clinical specimens showed that the kit B could be used to extract DNA from paraffin-embedded specimens,with the detection rate similar to that of kit A and freeze-thawing method.Conclusions The kit A could rapidly yield high-quality genomic DNA of Mycobacteria by repeated cleaning of columns,and may serve as the optimal method for scientific and clinical studies,and the kit B is suitable for extracting mycobacterial DNA from fresh tissue specimens besides paraffin-embedded specimens.