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 detect the plasma macrophage inflammatory protein (MIP) levels in patients with early Parkinson' s disease (PD) and to investigate whether plasma MIP was associated with motor and non-motor symptoms in early PD.Methods Fifty-nine patients with early idiopathic PD (Hoehn-Yahr Staging Scale from 1.0 to 2.5) treated in our hospital from January 28,2013 to September 30,2013 and 54 healthy controls were recruited.Plasma MIP-1α and MIP-1β levels were measured by enzyme-linked immunosorbent assay.Motor function was assessed by Unified Parkinson' s Disease Rating Scale Part Ⅲ and Hoehn-Yahr Staging Scale during “on” period.Total non-motor symptoms were assessed by Non-motor Symptoms Questionnaire.Cognitive dysfunction was assessed by Mini Mental State Examination.Autonotic dysfunction was assessed by Scales for Outcomes in Parkinson' s disease-Autonomic.Depression was assessed by Hamilton Depressive Scale (HAMD).Rapid eye movement (REM) sleep behavior disorder was assessed by REM sleep behavior disorder screening questionnaire (RBDSQ).Correlation between plasma MIP levels and scale scores was analyzed by Spearman rank correlation.Results Plasma MIP-1o and MIP-1β levels were not significantly different between early PD patients and healthy controls.However,plasma MIP-1 α level negatively correlated with depression (HAMD score,r =-0.520,P =0.027) and rapid eye movement sleep behavior disorder (RBDSQ score,r =-0.537,P =0.039).Conclusion MIP-1 α may be correlated with depression and RBD in early PD.

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.

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.

A case of benign follicular neoplasm-trichogerminoma-is reported.A 48-year-old man presented with a 10-year history of asymptomatic subcutaneous nodule in the chest.Histological examination revealed a well-circumscribed lesion composed of variously sized lobuli and cysts in the deep dermis and separated from the surrounding tissue by a fibrous capsule.Most lobuli consisted of concentrically arranged clear cells in the central area and basophilic cells in a palisade arrangement in the peripheral area.The tumor cells displayed a multi-directional differentiation toward hair bulb,inner root sheath,outer root sheath and infundibulum of hair follicles.Immunohistochemically,the tumor cells expressed AE1/AE3,CK5/6 and CK17,but were negative for CK20 or CK7.There was a sharp contrast in immunohistochemical findings between the central clear cells and peripheral basophilic cells.Based on the histological and immunohistochemical features,a diagnosis of trichogerminoma was made.

Objective To identify the outcomes and effect of applying the health check-up nursing quality control system developed as supervised by JCI quality assessment standards.Methods The monitoring system for nursing quality in the health check-up department is established within the framework of the hospital quality improvement committee and in line with characteristics of the department.Quality improvement tools may be called into play for analysis and decision making to revolve critical problems found in health check-up nursing,upgrading nursing quality and customer satisfaction.Results Significant rise of health check-up quality and customer satisfaction for nurses,and the nurses are trained in quality control knowledge and get further involved in quality management of their department.Conclusion Health check-up nursing management system under the JCI standard is conducive to raising the nursing quality,and helps nurses with problem analysis and solution.

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.

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.

Objective To construct the short hairpin RNA (shRNA)-expressing plasmid vectors specific for BRAF gene, and to test their effects in BRAF knockdown in human melanoma cell lines. Methods Two pairs of specific BRAF shRNA oligoes and a pair of randomly synthesized non-specific shRNA oligo were synthesized and inserted into plasmid pGenesil-1. Their fidelity was confirmed by double endonuclease digestion and sequencing. The constructed plasmids were transfected into human melanoma cell lines A375 and M14. The expression of BRAF mRNA and BRAF protein were detected by RT-PCR and Western blotting, respectively. Results The designed shRNA oligoes were precisely cloned into the plasmid pGenesil-1. The expression of BRAF mRNA and protein were down-regulated by specific plasmid braf 1 and braf 2, except to non-specific plasmid neg. The plasmid braf 1 was more effective, reducing BRAF gene expression by 90 per cent. Conclusions Plasmid mediated shRNA could successfully knockdown BRAF expression in human melanoma cells, and the suppression of the gene expression could maintain for 1 month at least.