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 compare and evaluate different DNA extraction methods for batches of bloodstain samples during the establishment of national DNA database. MethodsDNA was extracted from bloodstain samples collected for the establishment of DNA database using manual Chelex-100, Biomek 3000 automation workstation with Chelex-100 and DNA-IQTM magnetic bead method respectively, and the quantity of DNA was determined by the real-time fluorescent quantitative PCR technique. Genotyping was performed using the AmpFLSTR STR Identifiler kit and 3100 genetic analyzer. ResultsThe concentrations of DNA templates obtained by manual Chelex-100, automatic Chelex-100 and DNA-IQTM magnetic bead method were 0.593?0.131ng/?l, 0.579?0.096ng/?l and 0.447?0.056ng/?l respectively, and the success rates of genotyping with these DNA templates were 100%, 98.9% and 99.5% respectively. ConclusionThe automatic Chelex-100 method proposed by this study can be applied in the establishment of large-scale DNA databases.

ObjectiveTo investigate the intervention effects of moxibustion and moxa smoke on blood lipids,hepatic pathological changes and intrahepatocytic molecules related to cholesterol metabolism and analyze the regulating effects of moxibustion and moxa smoke on cholesterol metabolism and explore the mechanisms of actions of moxibustion and moxa smoke. MethodFifty-one 8-week-old ApoE-/-mice were randomized into model, moxa smoke and moxibustion groups, 17 mice each. Twenty C57BL/6 mice comprised a blank control group. The normal and model groups of mice were routinely grabbed and fastened. The moxa smoke group of mice was exposed to 10-15 mg/m3moxa smoke circumstances. The moxibustion group of mice was given moxibustion on point Guanyuan(CV4). All interventions were made 20 min daily, 6 times a week, for 12 consecutive weeks. Total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were measured using an automatic biochemical analyzer. Hepatic pathologic morphology was observed by HE staining. Hepatic CD36 and ABCA1 expressions were determined by immunohistochemical method.ResultIn the model group of mice, serum TG and LDL-C contents were significantly higher than in the normal group (P=0.003;P=0.001);HDL-C content was significantly lower than in the normal group (P=0.007); TC content had no significant difference compared with the normal group (P>0.05). In the moxibustion group of mice, serum TG and LDL-C contents were significantly lower than in the model group (P=0.03;P=0.001) and HDL-C content had no significant difference compared with the model group (P=0.11). In the moxa smoke group of mice, serum TG and LDL contents were significantly lower than in the model group (P=0.01;P=0.008) and HDL content had no significant difference compared with the model group (P=0.11). There were no significant differences in various blood lipid indicators between the moxibustion and moxa smoke groups (P>0.05). There were hepatic cell cord and sinusoid derangement and obvious hepatocytic swelling in the model group of mice. In the moxa smoke and moxibustion groups, hepatocytic swelling subsided significantly, and inflammatory cell infiltration reduced compared with the model group. In the model group,CD36 expression was significantly higher than in the normal group (P=0.004) and ABCA1 expression was significantly lower than in the normal group (P=0.001). In the moxibustion group, CD36 expression had no significant difference compared with the model group (P=0.09) and ABCA1 expression was significantly higher than in the model group (P=0.03). In the moxa smoke group, CD36 expression was significantly lower than in the normal group (P=0.02) and ABCA1 expression was significantly higher than in the model group (P=0.002). There were no significant differences in CD36 and ABCA1 expressions between the moxibustion and moxa smoke groups (P>0.05).ConclusionEarly moxibustion on point Guanyuan can regulate disorders of blood lipid metabolism, delay the occurrence of hepatic lesions and reduce intrahepatic accumulation of cholesterol to a certain extent in an ApoE-/-mouse model of atherosclerosis. That may be one of the mechanisms by which moxibustion therapy prevents atherosclerosis. Moxa smoke as the product of moxibustion is an effective factor in moxibustion producing a therapeutic effect.

ObjectiveTo investigate the effects of moxa smoke versus tobaccosmoke on autonomous behaviors and hippocampal expression of glial fibrillary acidic protein (GFAP) in apolipoprotein E gene knockout (ApoE-/-) mice.Method Thirteen 8-week-old C57BL/6 mice constituted a blank control group. Twenty-seven ApoE-/-mice of the same age were randomized into ApoE-/-model, moxa smoke and tobacco smoke groups. The tobacco smoke and moxa smoke groups of mice were exposed to smoke 5-15 mg/m3circumstances. Every group of mice was intervened in 20 min. daily, six days a week, for atotal of 12 weeks. A behavioral test was conducted in week 13. The animals were then sacrificed to take the materials. Hippocampal GFAP in the brain was measured by an immunohistochemical method.ResultAutonomous activities were significantly more in theblank group than in the model group (P<0.05) and significantly fewer in the tobacco smoke group than in the blank group (P<0.05). There was no statistically significant difference in autonomous activities between the tobacco smoke group and the model or moxa smoke group (P>0.05). Moving distance was longer in the moxa smoke group than in the model group (P<0.05). Standing-up number was smaller in the moxa smoke group than in the blank group (P<0.05). Integral optical density of GFAP immune reaction products in the hippocampus was significantlyhigher in the model group of mice than in the blank and moxa smoke groups (P<0.05). Hippocampal GFAP expression was significantly higher in the group of mice than in the moxa smoke and blank groups (P<0.05).Conclusion Moxa smoke can increase the excitability of central nervous system in mice and reduce hippocampal GFAP expression in a mice model of Alzheimer disease.

Objective To observe the effect of PM10 (inhalable particles) in moxa smoke on apoptosis of human lung adenocarcinoma cell line-A549 cells, and explore the possible mechanisms of inducing apoptosisMethods The A549 cells were studied in vitro experiment method, which were stained by Hoechest33258 staining method. Their morphological changes of apoptosis were observed by the fluorescence microscopy. The levels of intracellular Ca2+ and reactive oxygen species (ROS), and expression of NF-κB p65 were also measured.Results Some apoptotic cells were observed after treated with moxa smoke PM10 in the concerntration of 400μg/mL. After 4 hours intervention by moxa smoke PM10 in A549 cells, the intracellular Ca2+ level increased significantly (P<0.01). Compared with the blank control group, the expression of NF-κB p65 decreased significantly (P<0.05) after intervention of moxa smoke PM10 with different concerntrations for 4 hours. When the A549 cells were cultured with moxa smoke PM10 for 20 hours, the expression of p65 decreased significantly (P<0.05) in the concerntrations of 70, 280μg/mL, while there was no significant change in the concerntration of 140μg/mL (P>0.05). Compared with the blank control group, ROS level was significantly lower (P<0.05) in A549 cells after intervention of moxa smoke PM10.Conclusion PM10 in moxa smoke could induce apoptosis of A549 cells, could increase cytosolic Ca2+ level.

Objective To study the effect of Perfadex and Wisconsin Unviersity solution on the function of pulmonary artery endothelium.Methods Small lobe pulmonary arteries were dissected from nine porcine lungs.The artery from each lung was cut into six rings in 2mm.Two of them were randomly incubated in Krebs,Perfadex or Wisconsin Unviersity solution (UW solution) at 4℃ for 4 hours.Endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxation (percentage of-7.5 logM U46619 precontraction) induced by bradykinin or calcium ionophone A23187 in the present of indomethacin,L-NNA and oxyhemoglobin were measured at 37 ℃ in the organ chambers.Results Vasoconstriction induced by U46619 is no significant difference among three groups (P＞ 0.05).Compared with Krebs group,the relaxation induced by bradykinin or A23187 was significantly decreased in Perfadex group (P＜0.01),while there is no significant difference in UW group (P＞0.05).Conclusion Endothelium-derived hyperpolarizing factor (EDHF) plays an important role in endothelium-mediated relaxation of porcine pulmonary artery.EDHF-mediated relaxation is impaired when the lung preserved with Wisconsin Unviersity solution,wheras its function is not affected by Perfadex solution.

Objective To detect the expression of tissue inhibitor of metalloproteinase-4 (TIMP-4) in cutaneous malignant melanoma (CMM) tissue and to assess its relationship with melanoma proliferation,invasion and metastasis.Methods Western blot was conducted to measure the protein expression of TIMP-4 in five fresh lesional and paratumoral tissue specimens of CMM and three fresh tissue specimens of nevi.Immunohistochemistry was carried out to quantify the expression of TIMP-4,Ki-67,matrix metalloproteinase-2 (MMP-2),vascular endothelial growth factor (VEGF) and CD63 in paraffin-embedded tissue samples from 43 cases of CMM and 51 cases of nevi.The degree of malignancy of melanoma was evaluated in these lesions.Results Western blot analysis showed that the expression of TIMP-4 was significantly higher in 4 of 5 CMM tissue specimens than in corresponding paratumoral tissue specimens and nevus tissue specimens.Immunohistochemistry revealed that the expression rate of TIMP-4 was 86.04％ (37/43) in melanoma tissue,compared to 19.6％ (10/51) in nevus tissue (x2 =31.55,P ＜ 0.05).The expression of TIMP-4 increased sequentially from in situ melanoma to invasive and metastatic melanoma (rs =0.309,P ＜ 0.05).As far as CMM was concerned,the TIMP-4 expression was uncorrelated with any of the known prognostic variables including clinical stage,Clark level,Breslow depth,presence of ulcer,and Ki-67 expression (all P ＞ 0.05),but positively correlated with the expressions of VEGF (rs =0.345,P ＜ 0.05) and CD63 (rs =0.555,P ＜ 0.01).The median expression level of TIMP-4 was significantly higher in MMP-2-positive than in MMP-2-negative melanoma tissue samples (3 vs.0,P ＜ 0.01).Conclusions TIMP-4 protein is highly expressed in CMM tissue,which may be closely associated with the initiation and progression of CMM,especially with the metastasis of and angiogenesis in CMM.

Objective To measure the expression of hypoxia-inducible factor (HIF)-1α in acral malignant melanoma (MM) tissue and to investigate its relationship with the stem cell factor (SCF)/c-kit pathway.Methods Immunohistochemical staining was performed to measure the expression of HIF-1α in tissue specimens from lesions of 93 patients with acral MM,21 with non-acral MM,39 with acral melanocytic nevi,and from the normal acral skin of 15 healthy human controls.Meanwhile,the expression of c-kit was detected by immunohistochemical staining in the 93 acral MM tissue specimens.Statistical comparisons were carried out by chi-square test and Mann-Whitney U test.The relationship of HIF-1α expression with c-kit expression as well as tumor progression and staging was assessed by Spearman correlation analysis.Results Immunohistochemistry showed that the expression rate of HIF-1α was 87.10％ (81/93) in acral MM specimens,90.48％ (19/21) in non-acral MM specimens,15.38％ (6/39) in acral melanocytic nevus specimens,but 0 (0/15) in the normal acral skin specimens.The expression of HIF-1α was significantly higher in acral MM lesions than in normal acral skin and acral melanocytic nevus lesions (both P ＜ 0.01),and significantly different between acral MM and non-acral MM lesions (P ＜ 0.01).Moreover,HIF-1α expression was positively correlated with Clark level and Breslow depth of melanoma (rs =0.442,0.368,respectively,both P ＜ 0.01),with the progression of acral MM (from in situ to aggressive and metastatic MM) (rs =0.420,P ＜ 0.01),and with the expression of c-kit (rs =0.307,P ＜ 0.01).Conclusions HIF-1α is highly expressed in acral MM,positively correlated with the staging,progression and aggression of MM,and co-expressed with c-kit in acral MM tissue,suggesting that both HIF-1α and c-kit take part in the pathogenesis of acral MM.

Objective To investigate the diagnostic value of trans-gastric peritoneoscopy with technique of natural orifice transluminal endoscopic surgery(NOTES)for tuberculosis peritonitis.Methods Clinical data of 20 patients with tuberculosis peritonitis diagnosed by trans-gastric peritoneoscopy via NOTES were retrospectively analyzed.Results All diagnoses were confirmed by biopsy.The findings of peritoneoscopy were defined as miliary type with miliary nodes scattered in ascites and on peritoneum,adhesive type with thickening of peritoneum and adhesion between peritoneum and intestines,cheese-like type with parietal peritoneal ulcer and cheese-like substances,and mixed type with 2 or 3 of above mentioned types.Positive findings in other laboratory examinations were hemoglobin decrease in 10(50％)patients,blood sedimentation rate increase in 16(80％),C reactive protein increase in 13(65％),CA125 increase in 18(90％),and positive tuberculin test in 9(45％).Abnormal findings were detected by chest X-ray in 8(40％)patients,by abdominal ultrasonography examination in 2(10％),by abdominal CT in 7(35％),and by colonoscopy in 1(5％).No abnormal results were found in all patients in anti-tuberculosis antibody test,ascites bacteria culture and gastroscopy.Conclusion Trans-gastric peritoneoscopy via NOTES with biopsy is effective for diagnosis of tuberculosis peritonitis.

Objective To investigate the curative effect of sodium nitroprusside in the treatment of EV71-related pneumoedema/pneumorrhagia and hypotension in this study. Methods This was a retrospective study of a total 10 children with EV71-related pneumoedema/pneumorrhagia treated with sodium nitroprusside using case-control methods. The clinical manifestations and outcomes of the 10 children (present cohort) were compared with those of 8 children (past cohort) who had EV71-related pneumoedema/pneumorrhagia without sodium nitroprusside. Results Among these 10 patients,all were at the appearance of pneumoedema/pneumorrhagia, cardiac arrhythmia and hypotension. Two patients showed severe cardiac arrhythmia, two patients showed cardiac arrest for many times. All 10 patients were treated with mechanical ventilation and other conventional treatments. At the first time of the manifest of hypotension,sodium nitroprusside were put into practice in all 10 patients. Ten patients were treated with intravenously sodium nitroprusside in the stage of hypotension by maxi30 min ～ 1 h,the patients showed an improvement in vultus,pulse and peripheral circulation and decrease of heart rate and elevation of blood pressure after 30 min ～2 h,but at least 2～6 h later,the blood pressure tended to stabilize at normal standard. Conclusion Hypotension is the intensive stage in EV71-related hand,foot and mouth disease ,and the shock syndrome caused by acute left ventricular disorder is related to sympathetic nerve activity. Sodium nitroprusside can effectively reduce the cardiac afterload,and correct shock and improve the prognosis.