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 effect of phenylhexyle isothiocyanate (PHI) on demethylation and activation of transcription gene p15 in acute leukemia cell line Molt-4. Methods DNA sequencing and modified methylation specific PCR (MSP) were used to screen p15-M and p15-U mRNA after Moh-4 cells were treated with PHI. P15 mRNA was measured by RT-PCR. Pl5 protein was detected by Western blotting. Results Hypermethylation of gene pl5 was apparently attenuated and activation of transcription p15 gene was de novo after 5 days exposure to PHI. PHI enhanced both the expression of p15 mRNA and p15 protein in a concentration-dependent manner. The ratio of the gray scale of p15 mRNA strap was 0.17±0.12 in control, 0.29±0.14 in PHI 10 μmol/L, 0.55±0.07 in PHI 20 μmol/L, 0.93±0.13 in PHI 40 μmol/L. Conclusion PHI could active demethylation and transcription of gene p15.

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 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.

Objective:To investigate the metabolism changes in corpus striatum of hemiparkinsonian monkeys after chronic high frequency stimulation in subthalamic nucleus (STN) by SPECT and PET imaging. Methods: Two hemiparkinsonian monkeys, induced by unilateral internal carotid artery infusion of 1-methyl-4-phenyl-1,2,3,6-tetrahydropy-rindine (MPTP), were implanted with stimulation electrodes in STN of lesioned side according to stereotaxic atlas of the monkey brain. SPECT and PET examination were performed before and 1, 3 months after stimulation. Results: PD symptoms of monkeys, including rigidity, bradykinesia and gait abnormality, were significantly improved. SPECT imaging indicated that dopamine transporter (DAT) binding to corpus striatum in stimulated side increased and D 2 receptor (D 2 R) decreased to the level of unlesioned side after stimulating. PET imaging showed that right corpus striatum had lower density than left corpus striatum before stimulation. One month after chronic high frequency stimulation, bilateral corpus striatum had low density, but the right was slightly higher than that of the left;3 months after stimulation the right was obviously higher than the left . Conclusion: The symptoms of hemiparkinsonian monkeys can be obviously improved by STN deep brain stimulation(DBS). The increased DAT binding and glucose metabolism of corpus striatum in stimulated side and the decreased D 2 R binding after stimulation suggest that the activity of striatal dopaminergic system may be promoted by STN DBS.

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 determine the expression of Notch pathway receptors (Notch1 and Notch4) and ligands (Jagged1 and Dll4) in cutaneous malignant melanoma (CMM) tissues,and to preliminarily explore the role of the Notch signaling pathway in the pathogenesis of CMM.Methods Immunohistochemical study was performed to determine the expression pattern and intensity of Notch1,Notch4,Jagged1 and Dll4 in 40 paraffin-embedded CMM specimens and 15 paraffin-embedded pigmented nevus specimens.Statistical analysis was carried out by chi-square test and Spearman rank correlation analysis with the SPSS 21.0 software.Results Notchl was detected in 31 (77.5％) of 40 CMM specimens,as well as in 3 of 15 pigmented nevus specimens,and the positive rates significantly differed between the two groups (x2 =15.281,P ＜ 0.001).However,no significant difference in the expression intensity of Notch1 was observed between 18 in situ melanoma tissues and 22 invasive melanoma tissues (x2 =0.631,P =0.427).In addition,the positive rates of Notch4,Jagged1 and Dll4 were also significantly higher in the CMM group than those in the pigmented nevus group (all P ＜ 0.05),and the expression intensity of Notch4,Jagged1 and Dll4 significantly differed between in situ and invasive melanoma tissues (all P ＜ 0.05).In CMM tissues,the expression of Notch1 was positively correlated with that of Jagged1 (rs =0.350,P =0.027) and Dll4 (rs =0.562,P ＜ 0.001),while the expression of Jaggedl was negatively correlated with that of Dl14 (rs =-0.734,P ＜ 0.001).Conclusion Abnormality of the Notch signaling pathway may be involved in the pathogenesis of melanoma,but further researches are still needed to elucidate the detailed mechanism.

Objective To construct a eukaryotic expression plasmid carrying the PKCI-1/HINT1 gene,to investigate its expression in A375 melanoma cells after transfection,and to evaluate its effects on apoptosis and autophagy of A375 cells.Methods The PKCI-1/HINT1 gene sequence was amplified by reverse transcription PCR (RT-PCR) with total RNA extracted from A375 cells as the template,then inserted into the eukaryotic expression plasmid PCDNA3.1 (+) to construct a recombinant plasmid,PCDNA3.1 (+)-PKCI-1/HINT1.Some A375 cells were classified into two groups to be transiently transfected with the recombinant plasmid (PCDNA3.1 (+)-PKCI-1/HINT1 group) or the empty plasmid PCDNA3.1 (+) (control group).After additional 48-hour culture,RT-PCR and Western blot analysis were performed to quantify the mRNA and protein expressions of PKCI-1/HINT1 respectively,Hoechst 33342 staining was conducted to detect apoptosis of A375 cells,Western blot analysis to detect the expressions of intracellular caspase-3 and autophagy-associated protein beclin1,and cell autophagy was observed by using the green fluorescent protein (GFP)-microtubule-associated protein 1 light chain 3 (LC3) labelling method combined with confocal laser scanning microscopy.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of A375 cells at 24,48,72 and 96 hours after transfection.Results Enzyme digestion and sequencing analysis confirmed that the eukaryotic expression plasmid PCDNA3.1 (+)-PKCI-1/HINT1 was successfully constructed and effectively expressed in the transfected A375 cells.MTT assay showed that PKCI-1/HINT1 could obviously inhibit the proliferation of A375 cells,and the number of live cells was decreased by 17.0％,25.6％ and 29.4％ in the PCDNA3.1 (+)-PKCI-1/HINT1 group at 48,72 and 96 hours,respectively,compared with the control group (all P ＜ 0.05).Hoechest 33258 staining revealed that PKCI-1/HINT1 could promote the formation of apoptotic bodies in A375 cells.Confocal laser scanning microscopy demonstrated that the overexpression of PKCI-1/HINT1 increased GFP-LC3 puncta formation in A375 cells.In addition,Western blot analysis indicated that PKCI-1/HINT1 up-regulated the protein expressions of caspase-3 and beelin1 in A375 cells.Conclusions The eukaryotic expression plasmid PCDNA3.1 (+)-PKCI-1/HINT1 was successfully constructed,and PKCI-1/HINT1 could be effectively expressed in A375 cells.High-level expression of PKCI-1/HINT1 could suppress cellular proliferation,promote apoptosis,and induce autophagy,of A375 cells.

Objective To observe the effect of electroacupuncture and nerve and tendon gliding exercises on mild to moderate carpal tunnel syndrome.Methods Forty patients with mild to moderate carpal tunnel syndrome were randomly divided into an observation group and a control group,each of 20.Both groups were given neurotrophic drugs.The observation group was additionally provided with electroacupuncture and nerve and tendon gliding exercises,while the control group was given routine health education for 4 weeks.The Boston carpal tunnel syndrome questionnaire (BCTQ),a visual analogue scale (VAS),thumb and middle finger to wrist motor conduction velocity (MCV),sensory nerve action potentials (SNAPs),median nerve distal motor latency (DML) and the amplitude of compound muscle action potential (CMAP) of the musculi abductor pollicis brevis were evaluated.Results There were no significant differences in any of the measurements between the two groups before the treatment.After four weeks of treatment,significantly better improvement was observed in the average SCV of the thumb and wrist,the SCV of the digitus medius and wrist,the SNAP of the thumb and wrist,the SNAP of the digitus medius and wrist,the DML of the nervus medianus and the CMAP of the abductor pollicis brevis in the observation group compared to before the treatment and compared to the control group.Moreover,there was a significant decrease in the observation group's average BCTQ symptom score and function score and in their VAS ratings.However,in the control group significant differences were only found in the average SCV of the thumb and wrist,the DML of the nervus medianus and in their VAS ratings.Conclusions Electroacupuncture and nerve and tendon gliding exercises can significantly relieve symptoms and restore hand function.They are worth applying in clinical practice.