Objective To assess the clinicopathological and confocal microscopic features of porokeratosis.Methods This study included 186 patients with porokeratosis.The clinical and pathological findings from the patients were retrospectively analyzed.Confocal laser scanning microscopy(CLSM)was used to observe the lesions of disseminated superficial porokeratosis in 16 patients.Results Most of the patients had characteristic lesions of porokeratosis,i.e.,papules or plaques with a thread-like elevated border.Comoid lamella was observed in all of the cases,which was unassociated with sweat glands or hair follicles in most cases(171/186),and located in sweat pore or hair follicles in a few cases(15/186).There were dyskeratocytes as well as vacuolized and degenerated basal cells beneath the cornoid lamella.Varying amounts of lymphocytes and melanophages were observed in the superficial dermis.Amyloid was deposited in the papilla dermis in 2 cases.CLSM showed dyskeratocytes in a characteristic arcuate arrangement in spinous cell layer.Conclusions The CLSM images of porokeratosis are consistent with its histopathological manifestations,and CLSM may serve as a sensitive and specific noninvasive method for the diagnosis of porokeratosis.

Objective • To extract serum IgA1 from patients with IgA nephropathy (IgAN) (end stage renal disease vs long-term stable renal function) to explore the effect on proliferation rate of human mesangial cells (HMCs) and the level of transforming growth factor-β1 (TGF-β1). Methods • Nine patients with primary IgAN were divided into rapidly progressive group (n=6) and long-term stable group (n=3). Jacalin affinity chromatography and sephacryl S-200HR molecular sieves were used to distract serum IgA1. HMCs were cultured and co-cultivated with different IgA1 concentration (10, 50, 250 and 1 000 μg/mL)at point of 12 h and 24 h respectively. The proliferation rate was measured by cell counting kit-8 (CCK8). The expression of TGF-β1 mRNA was measured via quantitative real-time PCR (qRT-PCR). Enzyme-linked immunosorbent assay (ELISA) was used to detect TGF-β1 protein. Results • Serum IgA1 from IgAN patients with different renal functions (end stage renal disease vs long-term stable renal function) activated proliferation of HMC significantly, presenting with time-dependence and concentration-dependence. The highest value showed at 250 μg/mL and 1 000 μg/mL respectively. Serum IgA1 in two groups of patients statistically increased the expression of TGF-β1 mRNA and protein. In group with end stage renal disease, the summit stood at 10 μg/mL and started to decrease by degrees afterwards; while in group with long-term stable renal function, the level of TGF-β1 increased in a parallel manner with the serum IgA1. Conclusion • Serum IgA1 from IgAN patients with different renal functions (end stage renal disease vs long-term stable renal function) can both promote the proliferation of HMC. There is no dramatical difference observed with in10-50 μg/mL, but the IgA1 from group with end stage renal disease reveals a stronger effect on TGF-β1, in accordance with the pathological type of the patients (IgA sclerosis), suggesting the prognostic value of serum IgA.

OBJECTIVETo investigate the transduction ability of PEP-1-CAT fusion protein into human umbilical vein endothelial cell (HUVECs) and the effects on hydrogen-peroxide (H2O2)-induced oxidative stress injury in these cells.

METHODSWith the use of TA-cloning program and isocaudamer technique, the pET15b-PEP-1-CAT of prokaryotic expression plasmid was successfully constructed. The recombinant plasmid was transformed into E.coli BL21 (DE3) and the protein expression was induced by IPTG. The recombinant protein has an N-terminal His-tag which could be used to purify the target protein by affinity chromatography on a Ni2+-NTA-resin column. The fusion protein PEP-1-CAT was prepared and confirmed by specific enzyme activity in vitro. The purified PEP-1-CAT fusion protein was added on cultured HUVECs in vitro. The transduction ability of PEP-1-CAT fusion protein into cells was analyzed by Western blot and specific enzyme activity. The cells were treated with H2O2 (0.5 mmol/L) alone and in combination with PEP-1-CAT fusion protein for 4 h. Then, the cell viability, lactate dehydrogenase (LDH) and malondialdehyde (MDA) contents were measured.

RESULTSThe PEP-1-CAT fusion protein could be transduced into the cultured HUVECs in a dose- and time-dependent manner and be stable for at least 48 h. After H2O2 administration, cell viability was significantly reduced compared with control group (37.23%+/-5.68% vs. 100%, P<0.05), while LDH leakage (849.3 U/L+/-95.1 U/L) and MDA (8.23 nmol/L+/-1.58 nmol/L) content were significantly higher than that in control group (540.6 U/L+/-65.7 U/L and 2.46 nmol/L+/-1.42 nmol/L, respectively, all P<0.05). Preincubation with PEP-1-CAT proteins at various concentrations (0.25-2 micromol/L) significantly attenuated H2O2-induced cell injury.

CONCLUSIONThe PEP-1-CAT fusion protein could efficiently penetrate HUVECs and the transduced protein could attenuate cellular oxidative stress injury induced by H2O2. The PEP-1-CAT fusion protein might be a new strategy for preventing and treating oxidative stress induced diseases.

Catalase ; metabolism ; Cells, Cultured ; Cysteamine ; analogs & derivatives ; metabolism ; Endothelial Cells ; metabolism ; Humans ; Hydrogen Peroxide ; Oxidative Stress ; physiology ; Peptides ; metabolism ; Umbilical Veins ; cytology

Present work was designed to quantitatively evaluate the performance of diffusion-weighted magnetic resonance imaging (DWI) in the diagnosis of the presence of metastasis in lymph nodes (LNs).Eligible studies were identified from systematical PubMed and EMBASE searches.Data were extracted.Meta-analyses were performed to generate pooled sensitivity and specificity on the basis of per-node,per-lesion and per-patient,respectively.Fourteen publications (2458 LNs,404 lesions and 334 patients) were eligible.Per-node basis demonstrated the pooled sensitivity and specificity was 0.82 (P＜0.0001) and 0.90 (P＜0.0001),respectively.Per-lesion basis illustrated the pooled sensitivity and specificity was 0.73 (P=-0.0036) and 0.85 (P＜0.0001),respectively.Per-patient basis indicated the pooled sensitivity and specificity was 0.67 (P=0.0909) and 0.86 (P＜0.0001),respectively.In conclusion,DWI has rather a negative predictive value for the diagnosis of LN metastasis presence.The difference of the mean apparent diffusion coefficients between benign and malignant LNs is not yet stable.Therefore,the DWI technique has to be further improved.

Objective To monitor the levels of radioactive contamination due to the Japanese Fukushima nuclear accident in Beijing,so as to provide scientific technical information for government to draw effective controlling strategy and processing mechanism.Methods The system of nuclear emergency nmonitoring was started,then the radioactive contamination levels of atmosphere,rain water,surface water and vegetables in Beijing were detected according to the relative survey regulations and technology criterions.Results During the period from 15 to 41 d after the accident,obvious radioactive contamination was found in the atmosphere of Beijing.The maximum radioactivity concentration of 131I ( 5.89 mBq/m3 ) was detected at 22 d after the accident.The radioactivity concentrations of 137Cs and 134Cs were surveyed forming their corresponding peaks at 20 d after the accident,but they were one magnitude lower than the peak value of 131I at least.In addition,the gross β radioactivity level in the water of Chao Bai-he river was verified to be in the range of 0.314 - 0.602 Bq/L. Conclusions The radioactive contamination due to Fukushima nuclear accident has not done visible harm to the public health in Beijing,but monitoring should be continued to observe the long-term effect of the accident.

OBJECTIVETo study the effect of Banxia Houpu Decoction on a chronic mild stress model of depression and investigate the antidepressive mechanism.

METHODWith consumption of a 1% sucrose solution as an index, and by subjecting rats to a variety of mild stressors for a prolonged period of time, a chronic mild stress model was developed. The levels of the blood lipid were measured by blood lipid kits, the natural kill (NK) cell activity in the spleen was measured with the method of the enzyme lactate dehydrogenase (LDH) released, the superoxide dismutase (SOD) activity in red blood cell was assayed by the Autoxidation of Pyrogallol method, the NO synthase (NOS) activity in serum and tissue was measured by NOS kits, and the content of malondialdehyde(MDA) was measured by MDA kits.

RESULTBanxia Houpu Decoction significantly increased the consumption of sucrose solution, increased the level of the high density lipoprotein (HDL-C), decreased the level of the Triglyceride(TG) in serum, enhanced the activity of the NK in spleen, decreased the activity of the SOD in red blood and the activity of the NOS in serum and tissue, and reduced the content of MDA in tissue by effect on lipid Peroxidation in CNS model of depression.

CONCLUSIONBanxia Houpu Decoction has antidepressant effect in different ways.

Animals ; Antidepressive Agents ; pharmacology ; Depression ; immunology ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Killer Cells, Natural ; immunology ; Male ; Malondialdehyde ; blood ; metabolism ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stress, Psychological ; complications ; Sucrose ; metabolism ; Superoxide Dismutase ; metabolism ; Triglycerides ; blood

OBJECTIVETo observe the effect of intracerebroventricular injection of rAAV-HIF-1α on hippocampal neuronal apoptosis in a rat model of Alzheimer disease (AD).

METHODSThirty-two male SD rats (250-300 g) were randomized into 4 groups (n=8), including the normal control group without any treatment, AD model group with right intracerebroventricular injection of 2 µl Aβ25-35 (10 mg/m1), sham-operated group with right intracerebroventricular injection of 2 µl normal saline, and AD+ rAAV-HIF-1α group with right intracerebroventricular injection of 10 µl rAAV-HIF-1a (1×10¹² v.g./m1) one week after Aβ25-35 injection. The rats were sacrificed to detect the expression of HIF-1α and apoptosis of hippocampal neurons 5 weeks after Aβ25-35 or saline injection.

RESULTSWestern blotting showed that the expression of HIF-1α was significantly higher in AD+rAAV-HIF-1α group (451.59±34.39) than in normal control group (229.05±41.28) and sham-operated group (216.29±37.08) (P<0.05) without significant difference between the latter two groups. The apoptotic ratio of the hippocampal neurons was significantly higher in AD model group ([19.49±2.59]%) than in normal control group ([5.41±0.75]%) and sham-operated group ([5.28±0.66]%) in (P<0.05), and intracerebroventricular injection of rAAV-HIF-1α resulted in a significant reduction of the apoptotic ratio in the AD rats ([12.07±2.06]%) (P<0.05).

CONCLUSIONIntracerebroventricular injection of rAAV-HIF-1α can inhibit hippocampal neuronal apoptosis in the rat model of AD.

Alzheimer Disease ; metabolism ; therapy ; Animals ; Apoptosis ; Disease Models, Animal ; Hippocampus ; cytology ; Hypoxia-Inducible Factor 1, alpha Subunit ; administration & dosage ; genetics ; metabolism ; Lateral Ventricles ; Male ; Neurons ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley

The megakaryocyte and platelet lineage may be one of the major sites of human cytomegalovirus (HCMV) infection. However, whether HCMV aggravates apoptosis in normal megakaryocytes was not well investigated. Megakaryocytic cell line CHRF-288-11 and HCMV AD 169 strain were co-cultured in this study. PCR was used to detect the direct infection of the cells by HCMV IEA expression. The apoptotic cells were analyzed by morphologic observation, DNA ladder formation, annexin V/PI and PI assay with flow cytometry. The results showed that HCMV significantly inhibited the growth of CHRF cells in three different concentrations of viral infection groups (10(-3), 10(-2), 10(-1)). The viability levels in each infection groups were 77%, 73% and 68% respectively after incubation for 7 days, compared with 98% in the control group. Using annexin V/PI with flow cytometry, it was shown that the percentages of apoptotic cells viral infection in groups (10(-3), 10(-2), 10(-1)) were (21.3 +/- 2.49)%, (25.8 +/- 3.65)% and (31.4 +/- 3.91)% at 7 days after infection, while the control was (3.68 +/- 1.47)%. The apoptotic cells were further confirmed by morphologic observation and DNA ladder formation. Furthermore, PCR detection also showed the direct infection by identification of HCMV IEA expression in CHRF cells. This study suggested that HCMV could directly infect megakaryocytes and aggravated apoptosis in HCMV-infected megakaryocytes.
Apoptosis ; Cell Survival ; Cells, Cultured ; Cytomegalovirus ; pathogenicity ; DNA, Viral ; analysis ; Humans ; Megakaryocytes ; cytology ; virology ; Polymerase Chain Reaction

OBJECTIVETo investigate the penetrating ability of fusion protein PEP-1-EGFP with human umbilical vein endothelial cells.

METHODSTwo prokaryotic expression plasmids pET15b-EGFP and pET15b-PEP-1-EGFP were constructed and transformed into E. coli BL21 (DE3) to express EGFP and fusion protein PEP-1-EGFP, respectively. The expressed EGFP and PEP-1-EGFP were purified with Ni(2+) -resin affinity chromatography, and their capabilities of transduction into human umbilical vein endothelial cells were evaluated. The time- and dose-dependent transduction of the fusion protein PEP-1-EGFP and its stability in the human umbilical vein endothelial cells were observed. The toxicity of the fusion protein PEP-1-EGFP was detected by MTT method.

RESULTSEGFP failed to be transduced into human umbilical vein endothelial cells, whereas PEP-1-EGFP fusion protein was transduced into cells shortly in 5 minutes. Its transduction was time- and dose-dependent and the fluorescence in the cells were detected even 27 hours later. No cytotoxicity of the fusion protein PEP-1-EGFP to human umbilical vein endothelial cells was detected even when the dose reached up to 200 micromol/L.

CONCLUSIONPEP-1-EGFP fusion protein can efficiently transduce the target protein into human umbilical vein endothelial cells, which provides a basis for future researches on the transduction of antioxidant enzymes mediated by the cell-penetrating peptide, PEP-1, in ischemia-reperfusion injury therapy.

Cells, Cultured ; Cysteamine ; analogs & derivatives ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Green Fluorescent Proteins ; metabolism ; Humans ; Peptides ; metabolism ; Protein Transport ; Recombinant Fusion Proteins ; metabolism ; toxicity ; Umbilical Veins ; cytology

Purpose To investigate the clinicopathologic characteristics, diagnosis, differential diagnosis and prognosis of malignant solitary fibrous tumor/hemangiopericytoma ( SFT/HPC). Methods Sixteen cases of intracranial malignant SFT/HPC were retrospectively studied. The clinical data, imaging features, histopathological and immunohistochemical characteris-tics were analyzed. Results The 8 male and 8 female patients were between 31 and 71 years of age ( mean 51). The median age was 51 years (range, 31-71 years). 16 malignant SFT/HPC cases were originated from intracalvarium. The imaging features showed intracranial neoplasms with relatively clear surrounding boundaries. Microscopically spindle shaped cells were hypercel-lular, and exhibited≥5 mitoses per 10 HPF. Cytological atypia was mild. The clinicopathologic characteristics included pattern-less growth pattern, storiform or fascicular growth pattern, solita-ry fibrous tumor-like regions and hemangiopericytoma-like re-gions. Tnere were 2 cases with abundant papillary structure and 2 with sarcomatous structure, 2 with focal necrosis, 2 with inva-ded cerebral tissues, and 10 with invaded meninges. Immuno-histochemically, 93. 75% ( 15/16 ) cases were positive for STAT6, with 15/16 showing diffuse staining. 87. 5% (14/16) cases were positive for CD34, with 37. 5% (6/16) showing dif-fuse staining. 81. 25% (13/16) cases were positive for BCL-2. 68. 75% (11/16) cases were positive for CD99. The Ki-67 in-dex ranged from 5% to 40% . Sixteen patients were followed up for 1-64 months, and 7 patients ( 43. 75% ) had recurrences. Conclusion Malignant SFT/HPC shares malignant behaviours. STAT6 is a specific marker for the diagnosis of this tumor. The prognosis of malignant SFT/HPC is related to the extent of tumor excision and long-term follow-up.