Objective To assess whether anemia is an independent predictor of poor long-term outcome after percutaneous coronary intervention(PCI).Methods The second drug-eluting stent impact on revascularization registry(DESIRE-2)is a single-center registry of 6005 patients undergoing coronary revascularization from July 2003 to September.2005.We examined the clinical data and outcome of 3809 PCI patients based on hemoglobin(Hb)value before the interventional procedure.Patients were classified as anemia using the World Health Organization deftnition(＜120 g/L in women and＜130g/L in men). 744 of the 3809 patients were anemic.We compared the clinical features and prognosis of the patients with or without anemia.Results Anemic patients were older and had a higher percentage of comorbidities as compared with the nonanemic ones.When compared with nonanemic patients,anemic patients had higher mortality(4.7%VS 1.5%,P＜0.001)and higher major adverse event end points,including nonfatal myocardial infarction,stroke and revascularization(14.0%vs10.8%,P=0.014).After adjustment for comorbidities,anemia was associated with a higher risk of mortality after percutaneous coronary intervention(RR 2.216,95%CI 1.019-4.428;P=0.024).Conclusions Anemia is an independent predictor of mortality after PCI.Since PCI iS a common procedure and anemia is a frequent condition in the general population,strategies for the management of anemic PCI patients should be developed.

Objective To construct the eukaryotic expression plasmids of short hairpin RNA (shRNA) specific for mouse Bcl-2-assoeiated athanogene 1 (BAG-1) and to observe their inhibitory effects on the expression of BAG-1 gene in mouse melanoma B16FI0 ceils. Methods Plasmids named pRNAT-U6.1/Neo-BAG-1, were designed and constructed to target the mouse BAG-1 mRNA coding region. LipofectaminTM 2000 was used to transfect plasmids into BI6F10 cells. Negative plasmid-transfected and tmtransfected B16F10 cells served as negative and blank controls respectively. Forty-eight hours following transfection, G418 was used to select the resistant cells. The mRNA and protein expression of BAG-1 gene was measured by reverse transcription-PCR and Western blot respectively about 1 month after the transfection. Results The eukaryotic expression plasmids, pRNAT-U6.1/Neo-BAG-1, were constructed, and verified by restriction enzyme digestion and DNA sequencing. The transfection rate in B16F10 cells was 20% -30%. Compared with the blank control, the mRNA and protein expression of BAG-1 in BI6FI0 cells was significantly inhibited by BAG-1 shRNA (both P<0.05), and the inhibition rates were (77±4)% and (62 ±2)%, respectively. Conclusions These results indicate that the eukaryotic expression vectors containing shRNA against BAG-1 gene, pRNAT-U6.1/Neo-BAG-1, are successfully constructed, and can significantly inhibit the expression of BAG-1 gene in mouse melanoma B16F10 cells.

Objective To investigate the effect of microRNA-let-7a on apoptosis in melanoma cell line A375 and its mechanism. Methods The expression of microRNA-let-7a was detected in A375 cells and melanocytes by real-time PCR. Then, microRNA-let-7a mimics was transfected into A375 cells followed by the measurement of apoptosis and caspase-3 protein expression by flow cytometry and Western blotting, respectively. Results The expression level of microRNA-let-7a was reduced by 0.462 folds in A375 cells compared with melanocytes. The apoptosis rate was 47.4% in A375 cells transfected with microRNA-let-7a mimics, significantly higher than that in untransfected A375 cells (16.9%). A significant decline was observed in the expression of caspase-3 protein in A375 cells after transfection. Conclusion microRNA-let-7a can promote the apoptosis and downregulate caspase-3 protein expression, in A375 human malignant melanoma cells.

A 41-year-old female patient developed round,bright yellow patches on the left calvarial region without obvious precipitating factors 40 years prior to the presentation,which gradually grew to form plaques with age.Two years prior to the presentation,nipple-like lesions appeared in the calvarial and temporal region with an erythematous and wet surface; concurrently,black masses developed in the left temporal region and gradually enlarged with central ulceration but no subjective symptoms.At about 1 year of age,pitchy macules developed on the light tan patches located on the left jaw,posterior and anterior neck,trunk and upper limbs,and gradually increased in quantity and size with the involvement of the homolateral dorsal hand and gradual appearance of papules.Skin examination revealed two well-marginated,indurated,bright red neoplasms sized 3 cm × 2 cm and 2 cm × 1 cm respectively,with erosive and cauliflower-like surface; black or pink papules were scattered between these neoplasms.There was a ring-shaped black mass sized 1.5 cm × 1.5 cm in the left temporal region with central ulceration.Pitchy tough macules and papules were observed on the light tan patches located in the left cheek,lower mandible,posterior and anterior neck,protothorax,shoulder and back,upper limbs and dorsal hand.Based on the histopathology of multiple lesions,the cauliflower-like lesions on the head were diagnosed as syringocystadenoma papilliferum,the yellow plaques as syringocystadenoma papilliferum complicated by sebaceous adenoma,the black proliferative lesions in the temporal region as trichoblastoma accompanied by basal cell epithelioma,the black papuloid lesions and brown maculopapuloid lesions on the lower mandible as nevus spilus.The patient was diagnosed with skin adnexal tumor with multipotential differentiation (syringocystadenoma papilliferum,sebaceous adenoma,trichoblastoma and basal cell epithelioma)accompanied by nevus spilus.

Objective To evaluate the effect ofT-cell immunoglobulin and mucin domain-3 (TIM-3) on TRP-2180-188 peptide-stimulated murine spleen lymphocytes co-cultured with B16F10 murine melanoma cells.Methods A recombinant plasmid pFUSE-TIM-3-mIgG2Aae1-Fc2 encoding TIM-3 was constructed.Then,the recombinant plasmid and an empty plasmid pFUSE-mIgG2Aae1-Fc2 were transfected into human 293T epithelial cells followed by 48-hour culture for the preparation of supernatants containing TIM-3 and Ig-tail respectively.C57BL/6 mice were immunized with the TRP-2180-188 peptide vaccine for 4 sessions.One week after the last vaccination,C57BL/6 mice were sacrificed,and spleen lymphocytes were collected and then cultured with the TRP-21180-188 peptide and interleukin-2 (IL-2) for 5 days,with lymphocytes untreated with the TRP-2180-188 peptide or IL-2 serving as the control group.Mitomycin-treated B16F10 murine melanoma cells and TRP-2180-188 peptide-stimulated lymphocytes were co-cultured with the presence of supernatants of 293T cells that had been cultured for 48 hours (blank control group),TIM-3-containing supernatants (TIM-3 group) and Ig-tail-containing supernatants (negative control group) separately.After 24 and 48 hours of co-culture,cell counting kit-8 (CCK-8) assay was performed to estimate the proliferative activity of lymphocytes,enzyme-linked immunosorbent assay (ELISA) to determine the supernatant levels of interferon (INF)-γ and tumor necrosis factor (TNF)-α,flow cytometry to determine the percentage of CD8 + T cells in the co-culture system.Results Enzyme digestion and sequence analysis showed that the TIM-3 gene was successfully inserted into the eukaryotic expression plasmid.After 48-hour culture,TIM-3 and Ig-tail expressions were detected in the supernatants of 293T cells transfected with the recombinant plasmid and empty plasmid respectively.As CCK-8 assay showed,the proliferative activity of lymphocytes was significantly lower in the TIM-3 group than in the blank control group and negative control group after 24-and 48-hour culture (78.06％ ± 6.37％ vs.100.00％ ± 10.42％ and 108.70％ ± 9.90％ at 24 hours,42.93％ ± 5.93％ vs.100.00％ ± 6.24％ and 168.00％ ± 2.98％at 48 hours,all P ＜ 0.05),so was the ratio of cellular proliferative activity at 48 hours to that at 24 hours (all P ＜ 0.05).Compared with the blank control group and negative control group,the TIM-3 group showed significantly decreased supernatant levels of IFN-γ and TNF-α after 24-hour (IFN-γ:192.96 γ 5.05 ng/L vs.216.44 ± 7.85 ng/L and 223.67 ±7.79 ng/L,both P＜ 0.05;TNF-α:58.43 ± 0.26 ng/L vs.26.43 ± 0.01 ng/L and 86.85 ± 1.12 ng/L,both P＜ 0.05) and 48-hour culture (IFN-γ:54.95 ± 0.57 ng/L vs.230.06 ± 4.23 ng/L and 167.24 ± 3.33 ng/L,both P ＜ 0.05;TNF-α:30.23 ±0.26 ng/L vs.26.84 ± 0.20 ng/L and 45.34 ± 0.22 ng/L,both P ＜ 0.05).In addition,the median percentage of CD8+ T cells was significantly increased in the TIM-3 group compared with the blank control group and negative control group after 24-and 48-hour culture (3.30％ vs.0.421％ and 2.22％ at 24 hours,4.06％ vs.0.577％ and 0.691％ at 48 hours,all P＜ 0.05).Conclusion TIM-3 in vitro can suppress the proliferative activity of and secretion of IFN-γand TNF-α by lymphocytes,but increase the percentage of CD8 + T cells in the co-culture system of TRP-2180-188 peptide-stimulated lymphocytes and B16F10 cells.

A 59?year?old female patient, who received bilateral lower limb amputation 39 years ago, presented with eczematoid changes in both lower limbs for over 20 years, and with chronic granuloma?like lesions complicated by verrucous hyperplasia for more than 10 years. There were large areas of infiltrative and proliferative lesions with exudation and peripheral erythema at the amputation sites in both knee joints. The lesions were hard with tenderness on palpation. Microscopic examination of lesional scales with 10%KOH showed negative results for fungi. However, three times of culture on the Sabouraud dextrose agar(SDA)medium all grew the same kind of fungus, and the front side and reverse side of its filamentous colony were white and orange yellow respectively. Microculture showed that linear hyaline conidiophores came out from lageniform mother cells with conidia ascending alongside. The conidia looked like dark brown eye lens, with an equatorial germ slit. Based on these findings, this fungus was identified as Arthrinium phaeospermum. Periodic acid?Schiff (PAS) staining showed scattered hyphae in the stratum corneum. The internal transcribe spacer(ITS)sequence of the isolated fungus showed 99%consistency with that of Arthrinium phaeospermum. The patient was diagnosed with cutaneous Arthrinium phaeospermum infection, and treated with oral itraconazole capsules 200 mg/d for 16 days. One month later, follow?up showed satisfactory outcomes.

Objective To understand the application of drug eluting stent (DES) in renal functional insufficient patients, and to assess its safety and effectiveness, especially the occurrence of stent thrombosis(ST) after DES implantation and its related factors. Methods The subjects were all the patients underwent percutaneous coronary intervention (PCI) as well as at least one DES admitted to Beijing Anzhen Hospital consecutively from July 2003 to June 2005. All patients were divided into 2 groups: Group Ⅰ with normal or mild renal insufficiency (Ccr≥60 ml/min),and Group Ⅱ with moderate to severe renal functional insufficiency (Ccr ＜ 60 ml/min). All of the clinical, angiography and intervention data were recorded. ST was adjudicated by the definition of ARC Dublin. The rates of MACCE in hospital and during the follow-up between the 2 groups were compared. Results There were 2377 patients enrolled in the study, of which 2020 ( 85.0% ) patients presented Ccr ≥ 60 ml/min, and 357( 15.0% ) presented Ccr ＜ 60ml/min. The case fatality during follow-up in group Ⅰ was significantly higher than that in group Ⅱ (4. 5% vs. 1.2%, P ＜ 0. 001 ). However, the incidences of ST were not significantly different between each stage of disease( P ＞0. 05 ). The results from Cox regression showed that renal functional insufficiency was not a risk factor of death,whereas multivessel coronary artery disease [OR = 1. 929(95% CI: 1. 178 -3. 157),P =0. 009] ,diabetes [OR = 1. 914(95% CI:1. 055 -3. 470) ,P =0. 033] and age [OR = 1. 051 (95% CI:1. 005 -1. 099 ) ,P = 0. 030] were independent risk factor of death after DES implantation in patients with moderate to severe renal functional insufficiency. Conclusions Compared with normal renal function or mild renal patients, the longterm case fatality is higher in moderate and severe renal functional insufficiency patients. However, the higher case fatality does not due to the increase of ST.

A 16-year-old woman presented plaques on the left auricle and face over a period of 3 years. Fungal culture grew black-grey or dust velvety colony on Sabouraud's dextrose agar plate. A slide culture on potato dextrose agar plate showed conidiophores which were unbranched or occasionally loosely branched. The conidia were sympodial, zero- to two- septate, with rounded apices and truncated bases. The optimum growth temperature was 26℃ - 30℃. The fungus had the ability to liquefy glutin and hydrolyze starch. Anti-fungal susceptibility test showed the fungus was susceptible to itraconazole, terbinafine and amphoterecin B, but resistant to fluconazole. Cutaneous biopsy specimens revealed brown hyphae and budding yeast cells. The sequence of internal transcribed spacer (ITS) 1-ITS4 region of the isolate rDNA was assessed and compared against the Genebank databases. A 99% consistence was observed in the ITS sequence between clinical isolate and reference strain of Veronaea botryose Ciferri et Momtemartini. Based on the above findings, the mold was identified as Veronaea botryose Ciferri et Momtemartini. The lesions gradually subsided after 8-month treatment with oral itraconazole of 100 mg twice daily.

Objective To investigate the in vitro anti-proliferation effect of a histone deacetylase inhibitor,chidamide,on a cutaneous malignant melanoma cell line,A375.Methods Cultured A375 cells were treated with different concentrations of chidamide(5,10,50,100,500 μmol/L)and aichostatin A (TSA)(0.1,0.25,0.5,1.0 μmol/L),respectively,for various durations(24,48,72,96,120 hours).Subsequently,cell proliferation,apoptosis and cell cycle were detected by MTT assay,annexin Vfluorescein isothiocyanate and propidium iodide double staining,and DNA ploid analysis,respectively.Results The proliferation of A375 cells was inhibited in a dose-dependent manner by chidamide of 5-500μmol/L and TSA of 0.1-1 μmol/L,and in a time-dependent manner from 0 to 120 hours after the beginning of trealment with ehidamide of 5-500μmol/L and TSA of 0.25-1μmol/L.The 48-hour 50% growth inhibition concentration(IC50)of ehidamide and TSA on A375 cells was about 250 μmol/L and 0.7μmol/L,respectively.After 48-hour treatment,the apoptosis mte was 80.27%±3.06%,79.53%±5.70%,83.13%±6.90%in A375 cells treated with chidamide of 62.5,125,250 μmol/L,respectively,16.27%±2.46%,28.83%±2.55%,83.40%±8.65%in those treated with TSA of 0.175,0.35,0.7 μmol/L,respectively,10.43%±0.96%in ontreated cells;a statistical increase was noticed in chidamide-treated cells and TSA-treated cells vs.untreated cells(all P＜0.001).A positive correlation was observed between the apoptosis rate and concentrations of TSA(r=0.955,P=0.000).Cell cycle analysis indicated that treatment with chidamide induced cell cycle arrest in G0/G1 phase,with the cell proportion in G0/G1 phase being 76.30%±6.06%,82.79%±0.74%,88.91%±5.29%in A375 cells treated with chidamide of 62.5,125,250μmol/L,respectively,versus 38.73%±3.36%in untreated cells.While after 48-hour treatment with TSA of 0.35 and 0.7 μmol/L,the proportion of cells in G2/M phases was 25.15%±2.71%and 58.71%±3.45%,respectively,compared to 15.73%±0.23%in untreated cells(P＜0.01).Conclusion Chidamide and TSA could induce cell cycle arrest and apoptosis,as well as inhibit the growth of A375 ceils in vitro.

Objective To investigate the effect of PCI/CABG for the patients with coronary artery disease(CAD) with LVEF≤40%. Methods 196 CAD patients with LVEF≤40% were divided into PCI (n = 106) and CABG group (n=90). The clinical factors, revaseularization, medication, and in-hospital and follow-up malignant cardiological events(MACCE) were retrospectively compared,and the survival curve were drew by COX regression.Results Survival rate of the PCI group and CABG group in 30-day (96% vs. 94% ), 1-year(95 % vs. 92% ), and 2-year(93% vs. 87% )were similar. There was no MACCE within 30 days in the two groups(92% vs. 92% ) and 1-year survival rate without MACCE was similar in the two groups(86% vs. 88% ). CABG group had higher 2- year survival rate without MACCE than PCI group (83%vs. 72% ). But the COX survival curves of the two group were not signifi-cantly different (P=0. 662). Conclusion The patients with LVEF ≤ 40% undergoing PCI have short-term and long-term survival rate similar to those accepting CABG.