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Previous studies have linked increased frequency of glycosylphosphatidylinositol-anchor protein (GPI-AP) deficiency with genomic instability and the risk of carcinogenesis. However, the underlying mechanism is still not clear. A randomForest analysis of the gene expression array data from 55 MDS patients (GSE4619) demonstrated a significant (p = 0.0007) correlation (Pearson r =-0.4068) between GPI-anchor biosynthesis gene expression and genomic instability, in which PIGN, a gene participating in GPI-AP biosynthesis, was ranked as the third most important in predicting risk of MDS progression. Furthermore, we observed that PIGN gene expression aberrations (increased transcriptional activity but diminished to no protein production) were associated with increased frequency of GPI-AP deficiency in leukemic cells during leukemic transformation/progression. PIGN gene expression aberrations were attributed to partial intron retentions between exons 14 and 15 resulting in frameshifts and premature termination which were confirmed by examining the RNA-seq data from a group of AML patients (phs001027.v1.p1). PIGN gene expression aberration correlated with the elevation of genomic instability marker expression that was independent of the TP53 regulatory pathway. Suppression/elimination of PIGN protein expression caused a similar pattern of genomic instability that was rescued by PIGN restoration. Finally, we found that PIGN bound to the spindle assembly checkpoint protein, MAD1, and regulated its expression during the cell cycle. In conclusion, PIGN gene is crucial in regulating mitotic integrity to maintain chromosomal stability and prevents leukemic transformation/progression.

Background: Ibrutinib, idelalisib, and venetoclax are approved for treating CLL patients in the United States. However, there is no guidance as to their optimal sequence., Patients and Methods: We conducted a multicenter, retrospective analysis of CLL patients treated with kinase inhibitors (KIs) or venetoclax. We examined demographics, discontinuation reasons, overall response rates (ORR), survival, and post-KI salvage strategies. Primary endpoint was progression-free survival (PFS)., Results: A total of 683 patients were identified. Baseline characteristics were similar in the ibrutinib and idelalisib groups. ORR to ibrutinib and idelalisib as first KI was 69% and 81%, respectively. With a median follow-up of 17 months (range 1-60), median PFS and OS for the entire cohort were 35 months and not reached. Patients treated with ibrutinib (versus idelalisib) as first KI had a significantly better PFS in all settings; front-line [hazard ratios (HR) 2.8, CI 1.3-6.3, P = 0.01], relapsed-refractory (HR 2.8, CI 1.9-4.1, P < 0.001), del17p (HR 2.0, CI 1.2-3.4, P = 0.008), and complex karyotype (HR 2.5, CI 1.2-5.2, P = 0.02). At the time of initial KI failure, use of an alternate KI or venetoclax had a superior PFS when compared with chemoimmunotherapy. Furthermore, patients who discontinued ibrutinib due to progression or toxicity had marginally improved outcomes if they received venetoclax (ORR 79%) versus idelalisib (ORR 46%) (PFS HR .6, CI.3-1.0, P = 0.06)., Conclusions: In the largest real-world experience of novel agents in CLL, ibrutinib appears superior to idelalisib as first KI. Furthermore, in the setting of KI failure, alternate KI or venetoclax therapy appear superior to chemoimmunotherapy combinations. The use of venetoclax upon ibrutinib failure might be superior to idelalisib. These data support the need for trials testing sequencing strategies to optimize treatment algorithms., (© The Author 2017. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Primary myelofibrosis is a unique entity among BCR-ABL-negative myeloproliferative diseases, manifesting as bone marrow fibrosis and pancytopenia. Considerable evidence indicates that genetic and epigenetic abnormalities can result in defective clonal hematopoietic stem cell proliferation in addition to bone marrow microenvironment alteration. The "bad seeds in bad soil" theory illustrates the orchestrating efforts of hematopoietic stem cells, stromal cells, and their surrounding signaling molecules in myelofibrosis progression and malignancy transformation, though the exact mechanism of myelofibrosis is still not clear. This study reviews current concepts and questions regarding the pathogenesis of primary myelofibrosis and discusses the emerging targeted therapy aimed at restoring normal bone marrow environment and halting bone marrow fibrotic deterioration.

B-cell receptor kinase inhibitor (KI) therapy represents a paradigm shift in chronic lymphocytic leukemia (CLL) management, but data on practice patterns after KI discontinuation and optimal sequencing are limited. We conducted a multicenter, retrospective, comprehensive analysis on 178 patients with CLL (ibrutinib = 143; idelalisib = 35) who discontinued KI therapy. We examined responses, toxicity, post-KI therapies, and overall survival (OS). Patients had a median of 3 prior therapies (range 0-11); del17p (34%), p53 mutation (27%), del11q (33%), and complex karyotype (29%). Overall response rate (ORR) to first KI was 62% (complete response 14%). The most common reasons for KI discontinuation were toxicity (51%), CLL progression (29%), and Richter transformation (RT) (8%). Median progression-free survival (PFS) and OS from KI initiation were 10.5 and 29 months, respectively. Notably, initial KI choice did not impact PFS or OS; however, RT portended significantly inferior OS (P = .0007). One hundred fourteen patients received subsequent salvage therapy following KI discontinuation with an ORR to subsequent KI at 50% and a median PFS of 11.9 months. Median PFS in KI-intolerant patients treated with an alternate KI was not reached vs 7 months for patients with CLL progression. In summary, these data demonstrate that toxicity was the most common reason for KI discontinuation, that patients who discontinue KI due to toxicity can respond to an alternate KI, and that these responses may be durable. This trial was registered at www.clinicaltrials.gov as #NCT02717611 and #NCT02742090., (© 2016 by The American Society of Hematology.)

Croton (Codiaeum variegatum (Linn.) var. pictum (Lodd.)) is an ornamental plant commonly grown in southern China. In March 2014, severe powdery mildew infections were observed on crotons in gardens of Hainan University (20.1°N and 110.3°E), Haikou, Hainan province. Disease incidence was estimated in a random batch of 100 plants in three replicates, with the average value approaching 80%. Symptoms first appeared as white circular patches on the adaxial surface and expanded to the abaxial surface, petioles, and stems. The top leaves were the most affected. Upper surfaces of the infected leaves were covered by white, dense mycelia. As the disease progressed, infected leaves turned purple on the lower surfaces and curly before becoming necrotic and abscising from the plant. Powdery mildew was more severe in shaded environments, especially during rainy or foggy weather in early spring. Two hundred conidiophores and conidia were observed microscopically. The conidiophores were straight or occasionally flexuous, 62.3 to 127.6 × 6.2 to 10.2 μm, consisting of two to three straight cells. Conidia were born in solitary on the top of conidiophores. Conidia were hyaline, ellipsoidal, 26.4 to 42.2 × 11.7 to 23.4 μm (average 32.5 × 16.5 μm), contained no distinct fibrosin bodies, and produced a subterminal germ tube. The wrinkling pattern of the outer walls of older conidia was angular or reticulated. Appressoria were single and multilobed. Cleistothecia were not observed. Based on morphological characteristics, the fungus was identified as Oidium neolycopersici (2), which was recently renamed Pseudoidium neolycopersici (L. Kiss) (3). The identity was confirmed by sequence analysis. Genomic DNA was extracted from the foliar powdery mildew colonies using Chelex-100 (Bio-Rad, Shanghai, China). The rDNA internal transcribed spacer (ITS) region was amplified with primers ITS1 and ITS4 (5). The ITS sequence of the representative isolates C01 (GenBank Accession No. KJ890378.1) and four other powdery mildew samples collected from crotons in Hainan University was 100% identical to that of P. neolycopersici isolates from tomato plants such as JQ972700 and AB163927. Inoculations were made by gently pressing diseased leaves onto leaves of five healthy plants of croton and tomato ('Money maker'). Five non-inoculated croton and tomato plants served as controls. Inoculated and non-inoculated plants were maintained in an incubator at 25°C with a 12-h photoperiod. After eight days, typical powdery mildew symptoms developed on 93% of the inoculated plants, while no symptom developed on the non-inoculated plants. The pathogenicity tests were repeated three times. The same fungus was always re-isolated from the diseased tissue according to Koch's postulates. The pathogenicity tests further confirmed that the pathogen from crotons is P. neolycopersici (Basionym. Oidium neolycopersici (KJ890378.1)), which is commonly known as the tomato powdery mildew. P. neolycopersici is also a pathogen of Normania triphylla (1) and papaya (4). To our knowledge, this is the first report of P. neolycopersici infecting croton. The avenue of this pathogen entering gardens of Hainan University remains unknown. The gardens are located far away from tomato farms. Also no symptom of powdery mildew on croton was observed during surveys in other locations in Haikou. The origin of the pathogen warrants additional research. References: (1) D. Delmail et al. Mycotaxon 113:269, 2010. (2) L. Kiss et al. Mycol. Res. 105:684, 2001. (3) L. Kiss et al. Mycol. Res. 115:612, 2011. (4) J. G. Tsay et al. Plant Dis. 95:1188, 2011. (5) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Cotton (Gossypium hirsutum L.) is one of the most important economic crops in China and fungal diseases are the major limiting factors in its production. In September 2013, cotton plants infected with leaf spots were observed in Sanya, Hainan Province, China. Initial symptoms developed as brick-red dots that led to the formation of irregular to circular lesions with gray centers surrounded by brown borders. Individual leaf spots formed concentric rings of alternating light and dark brown bands. Leaf tissue segments collected from the border between symptomatic and healthy tissue were surface disinfested in 75% ethanol for 1 min, then rinsed three times in sterile water with streptomycin sulfate. Fungal isolates obtained from these segments were purified by the single spore technique on potato dextrose agar (PDA) at 28°C. The initial color of the colonies was olivaceous, turning dark brown after 5 days. Conidiophores were scattered or clustered, brown, straight to curved, unbranched, and glabrous. Conidia had 4 to 12 pseudosepta and were 56 to 230 μm long and 5 to 15 μm wide, brown, straight to slightly curved, obclavate to cylindrical, glabrous, and apex obtuse. These characteristics were consistent with the description of Corynespora cassiicola (Berk. & M.A. Curtis.) C.T. Wei (3). A pathogenicity test was conducted with the four isolates on detached young cotton leaves (two to four true leaf stage). For each isolate, three slightly wounded and three unwounded leaves were inoculated with 5.5-mm-diameter mycelial plugs. For the control treatment, wounded and unwounded leaves were mock inoculated with sterile PDA plugs of the same size. The inoculated leaves were placed in a moist chamber and incubated with a 12-h photoperiod at 28°C. Necrotic lesions appeared on wounded spots after 2 days of incubation and on unwounded leaves 3 days after incubation. All symptoms were similar to those observed in the field. Symptoms were not observed on control leaves. The same fungus was always re-isolated from the diseased tissue according to Koch's postulates. To confirm the identity of the pathogen, DNA was extracted from a 1-week-old culture grown on PDA and the internal transcribed spacer region (ITS) of one isolate (GenBank Accession No. KF924624) was amplified using primers ITS1 and ITS4 (4) and sequenced. BLAST search in GenBank revealed 100% homology with sequences of C. cassiicola (EU364535.1, EU364536.1, FJ852574.1, and FJ852575.1). Based on the symptoms, fungal morphology, ITS sequence comparison, and pathogenicity test, this fungus was identified as C. cassiicola. Target spot of cotton associated with C. cassiicola has been reported in Georgia (2) and Alabama (1). To our knowledge, this is the first report that C. cassiicola can infect cotton in China inducing target spot of cotton (2). This report will establish a foundation for further study of C. cassiicola to aid disease measurement and control. References: (1) K. N. Conner et al. Plant Dis. 97:1379, 2013. (2) A. M. Fulmer et al. Plant Dis. 96:1066, 2012. (3) J. Y. Lu. Page 407 in: Plant Pathogenic Mycology. China Agricultural Press, Beijing, 2000. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Objective: To compare efficacy differences between fire filiform needle combined with mild moxibustion and gabapentin combined with sham acupuncture for postherpetic neuralgia (PHN)., Methods: One hundred cases of PHN were randomly divided into a needle group and a medicine group, 50 cases in each one. In the needle group, pricking method of fire filiform needle was given at the Ashi points, and then mild moxibustion was applied for 15 min. In the medicine group, the oral administration of gabapentin capsule and sham acupuncture at non-acupoints in the distal end of lesions were applied. The treatment was required for 21 days in both groups. The visual analogue score (VAS) was recorded before treatment and on the 1st day, 2nd day, 3rd day, 6th day, 9th day and 12th day of treatment. The most severity of pain within last 24 h, preset severity of pain, immediate analgesia effect and starting time of pain relief were observed, also the efficacy was assessed and improvement of symptoms was observed in the follow-up visit., Results: The total effective rate was 94.0% (47/50) in the fire filiform needle group, which was superior to 86.0% (43/50) in the medicine group (P < 0.05). Compared with medicine group, the VAS of the most severity of pain within last 24 h was obviously reduced after the 2nd treatment in the fire filiform needle group while that of present severity of pain was relieved after the 1st treatment (both P < 0.05). The immediate analgesia effect in the fire filiform needle group was obviously superior to that in the medicine group in the first three times of treatment (all P < 0.05). The average time of pain relief was (3.91 +/- 0.82) days in the fire filiform needle group, which was significantly earlier to (6.53 +/- 1.13) days in the medicine group (P < 0.05). 26 cases were cured in the fire filiform needle group in the follow-up visit, which was superior to 2 cases in the medicine group (P < 0.05). The improvement of VAS, pain range and sleep quality in the needle group were also superior to those in the medicine group (all P < 0.05). The direct medical cost in the fire filiform needle group was (232.32 +/- 48.108) yuan, which was significantly lower than (466.00 +/- 41.09) yuan in the medicine group (P < 0.05). There was only one case of adverse effect in the medicine group during the treatment., Conclusion: The fire filiform needle combined with mild moxibustion could obviously relieve the pain in PHN patients, which has superior immediate analgesia effect and pain relieving time compared with gabapentin, which also has less adverse effects and cheap cost.

To establish a proteomic reference map of Musa acuminate Colla (banana) leaf, we separated and identified leaf proteins using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). Tryptic digests of 44 spots were subjected to peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS. Three spots that were not identified by MALDI-TOF MS analysis were identified by searching against the NCBInr, SwissProt, and expressed sequence tag (EST) databases. We identified 41 unique proteins. The majority of the identified leaf proteins were found to be involved in energy metabolism. The results indicate that 2D-PAGE is a sensitive and powerful technique for the separation and identification of Musa leaf proteins. A summary of the identified proteins and their putative functions is discussed.

The interaction between ovalbumin (OVA) and three purine alkaloids (caffeine, theophylline and diprophylline) was investigated by the aid of intrinsic and synchronous fluorescence, ultraviolet-vis absorbance, resonance light-scattering spectra and three-dimensional fluorescence spectra techniques. Results showed that the formation of complexes gave rise to the fluorescence quenching of OVA by caffeine, theophylline, and diprophylline. Static quenching was confirmed to results in the fluorescence quenching. The binding site number n, apparent binding constant KA and corresponding thermodynamic parameters were measured at different temperatures. The binding process was spontaneous molecular interaction procedures in which both enthalpy and Gibbs free energy decreased. Van der Waals forces and hydrogen bond played a major role in stabilizing the complex. The comparison between caffeine, theophylline, and diprophylline was made, and thermodynamic results showed that diprophylline was the strongest quencher and bound to OVA with the highest affinity among three compounds. The influence of molecular structure on the binding aspects was reported.

Impacts of surface wind direction, surface wind speed, surface air temperature and sunshine hours on the CH4 concentration at Lin'an regional atmospheric background station were studied based on the results from Jan. 2009 to Dec. 2011. The results revealed that the diurnal variation of atmospheric CH4 concentration presented a single-peak curve at Lin'an regional background station. The diurnal amplitude varied from 19.0 x 10(-9) to 74.7 x 10(-9), with the lowest value observed in the afternoon and the highest at dawn. The monthly mean CH4 concentrations varied from 1955.7 x 10(-9) to 2036.2 x 10(-9), with the highest concentration observed in autumn and the lowest in spring. The wind directions NE-SSE could induce higher CH4 concentrations while SW-NNW wind directions had negative effects on the observed results. The CH4 concentration turned out to be lower with higher surface wind speed. With the increase of surface air temperature or sunshine hours, the CH4 concentration went up first till reaching a peak, and then decreased.

Background: Patients with paroxysmal nocturnal hemoglobinuria harbor clonal glycosylphosphatidylinositol-anchor deficient cells arising from a multipotent hematopoietic stem cell acquiring a PIG-A mutation. Many patients with aplastic anemia and myelodysplastic syndromes also harbor small populations of glycosylphosphatidylinositol-anchor deficient cells. Patients with aplastic anemia often evolve into paroxysmal nocturnal hemoglobinuria; however, myelodysplastic syndromes seldom evolve into paroxysmal nocturnal hemoglobinuria. Here, we investigate the origin and clonality of small glycosylphosphatidylinositol-anchor deficient cell populations in aplastic anemia and myelodysplastic syndromes., Design and Methods: We used peripheral blood flow cytometry to identify glycosylphosphatidylinositol-anchor deficient blood cells, a proaerolysin-resistant colony forming cell assay to select glycosylphosphatidylinositol-anchor deficient progenitor cells, a novel T-lymphocyte enrichment culture assay with proaerolysin selection to expand glycosylphosphatidylinositol-anchor deficient T lymphocytes, and PIG-A gene sequencing assays to identify and analyze PIG-A mutations in patients with aplastic anemia and myelodysplastic syndromes., Results: Twelve of 15 aplastic anemia patients were found to harbor a small population of glycosylphosphatidylinositol-anchor deficient granulocytes; 11 of them were found to harbor a small population of glycosylphosphatidylinositol-anchor deficient erythrocytes, 10 patients were detected to harbor glycosylphosphatidylinositol-anchor deficient T lymphocytes, and 3 of them were detected only after T-lymphocyte enrichment in proaerolysin selection. PIG-A mutation analyses on 3 patients showed that all of them harbored a matching PIG-A mutation between CFU-GM and enriched T lymphocytes. Two of 26 myelodysplastic syndromes were found to harbor small populations of glycosylphosphatidylinositol-anchor deficient granulocytes and erythrocytes transiently. Bone marrow derived CD34(+) cells from 4 patients grew proaerolysin-resistant colony forming cells bearing PIG-A mutations. No glycosylphosphatidylinositol-anchor deficient T lymphocytes were detected in myelodysplastic syndrome patients., Conclusions: In contrast to aplastic anemia and paroxysmal nocturnal hemoglobinuria, where PIG-A mutations arise from multipotent hematopoietic stem cells, glycosylphosphatidylinositol-anchor deficient cells in myelodysplastic syndromes appear to arise from more committed progenitors.

Commercial banana varieties are highly susceptible to fungal pathogens, as well as bacterial pathogens, nematodes, viruses, and insect pests. The largest known family of plant resistance genes encodes proteins with nucleotide-binding site (NBS) and C-terminal leucine-rich repeat (LRR) domains. Conserved motifs in such genes in diverse plant species offer a means for the isolation of candidate genes in banana that may be involved in plant defense. Six degenerate PCR primers were designed to target NBS and additional domains were tested on commercial banana species Musa acuminata subsp malaccensis and the Musa AAB Group propagated in vitro and plants maintained in a greenhouse. Total DNA was isolated by a modified CTAB extraction technique. Four resistance gene analogs were amplified and deposited in GenBank and assigned numbers HQ199833-HQ199836. The predicted amino acid sequences compared to the amino acid sequences of known resistance genes (MRGL1, MRGL2, MRGL3, and MRGL4) revealed significant sequence similarity. The presence of consensus domains, namely kinase-1a, kinase-2 and hydrophobic domain, provided evidence that the cloned sequences belong to the typical non-Toll/interleukin-1 receptor-like domain NBS-LRR gene family.

Characteristics of Atmospheric CO2 concentration obtained by Flask measurements were analyzed at Lin'an regional background station from August 2006 to July 2009. According to the simulation results of carbon tracking model, the impact of carbon sources and sinks on CO2 concentration was evaluated in Yangtze River Delta. The results revealed that atmospheric CO2 concentrations at Lin'an regional background station were between 368.3 x 10(-6) and 414.8 x 10(-6). The CO2 concentration varied as seasons change, with maximum in winter and minimum in summer; the annual difference was about 20.5 x 10(-6). The long-term trend of CO2 concentration showed rapid growth year by year; the average growth rate was about 3.2 x 10(-6)/a. CO2 flux of Yangtze River Delta was mainly contributed by fossil fuel burning, terrestrial biosphere exchange and ocean exchange, while the contribution of fire emission was small. CO2 flux from fossil fuel burning played an important role in carbon source; terrestrial biosphere and ocean were important carbon sinks in this area. Seasonal variations of CO2 concentration at Lin'an regional background station were consistent with CO2 fluxes from fossil fuel burning and terrestrial biosphere exchange.

Objective:  To investigate the natural history of paroxysmal nocturnal hemoglobinuria (PNH) clones in patients with acquired aplastic anemia (AA)., Patients and Methods:  Twenty-seven patients with AA and a detectable PNH clone were monitored for a median of 5.7 years (range 1.5-11.5 years). Twenty-two patients received high-dose cyclophosphamide (HiCy) therapy. The erythrocyte and granulocyte PNH clone sizes were measured using flow cytometry and analyzed via CellQuest software. PE-conjugated anti-glycophorin A, anti-CD15, FITC-conjugated anti-CD59, and FLAER staining were used to define glycosylphosphatidylinositol-AP-deficient cells., Results: We found a linear relationship between PNH clone size and the development of intravascular hemolysis, assessed by lactate dehydrogenase (LDH) values (Pearson correlation coefficient = 0.80, P < 0.001 for erythrocyte PNH clones; and Pearson correlation coefficient = 0.73, P < 0.0001 for granulocyte PNH clones). An erythrocyte PNH size of 3-5% and granulocyte PNH size of 23% were the thresholds to predict hemolysis as measured by an elevated LDH (receiver operating characteristic analyses with AUC = 0.96 for erythrocyte PNH clone sizes and AUC = 0.88 for granulocyte PNH clone sizes). Patients with small (≤15%) initial PNH clone sizes were less likely to develop an elevated LDH (mean ± SD: 236.9 ± 109.9 vs. 423.1 ± 248.8; P = 0.02). Over time, the PNH clone sizes remained stable in 25.9% of patients; 48.1% experienced a rise in the PNH clone size; and 25.9% experienced a decrease., Conclusion: The risk of developing clinically significant PNH after HiCy therapy appears to be low in AA patients with PNH clones, especially for those with small initial PNH clones and for those who respond to HiCy therapy., (© 2011 John Wiley & Sons A/S.)

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare hematologic disease that presents with protean manifestations. Clinical and laboratory investigation over the past 25 years has uncovered most of the basic science underpinnings of PNH and has led to the development of a highly effective targeted therapy. PNH originates from a multipotent hematopoietic stem cell (HSC) that acquires a somatic mutation in a gene called phosphatidylinositol glycan anchor biosynthesis, class A (PIG-A). The PIG-A gene is required for the first step in glycosylphosphatidylinositol (GPI) anchor biosynthesis. Failure to synthesize GPI anchors leads to an absence of all proteins that utilize GPI to attach to the plasma membrane. Two GPI-anchor proteins, CD55 and CD59, are complement regulatory proteins; their absence on the surface of PNH cells leads to complement-mediated hemolysis. The release of free hemoglobin leads to scavenging of nitric oxide and contributes to many clinical manifestations, including esophageal spasm, fatigue, and possibly thrombosis. Aerolysin is a pore-forming toxin that binds GPI-anchored proteins and kills normal cells, but not PNH cells. A fluorescinated aerolysin variant (FLAER) binds GPI-anchor and serves as a novel reagent diagnosing PNH. Eculizumab, a humanized monoclonal antibody against C5, is the first effective drug therapy for PNH., (© 2011 Wiley Periodicals, Inc.)

Gallbladder carcinosarcoma is a rare gastrointestinal tract malignant tumour, which contains both epithelial cancer component and mesenchymal sarcoma component. Because of its unique anatomic location and unspecific medical presentation, preoperative diagnosis is difficult. The prognosis of gallbladder carcinoma is poor with median survival time of 5.5 months. T- and N-staging system has no role in cancer prognostic stratification. Currently, we still have limited experience in managing this form of notorious cancer. Surgical resection is the common practice in treating gallbladder carcinoma though recurrence rate is high (80%). Here, we report a 59-year-old female with new diagnosed gallbladder carcinosarcoma. She was found to have a carcinosarcoma masse at gallbladder extending into proximal bile ducts with no metastatic lesion. She underwent a radical cholangiocholecystocholedochectomy and Roux-en-Y hepatocholangioduodenostomy. Postsurgery, she received six cycles of adjuvant chemotherapy consisting of oxaliplatin and 5-fluorouracil. She maintains in complete remission 6 months after adjuvant chemotherapy.

Background: Kell and XK, two distinct red blood cell membrane proteins, are linked by a disulfide bond and form the Kell blood group complex. Kell surface antigens are expressed early during erythropoiesis but the onset of expression of XK which carries the Kx antigen is unknown., Study Design and Methods: To determine whether Kell and XK are synchronously expressed, sorted human hematopoietic progenitor cells and mouse progenitor cells of defined lineage were studied. To determine the onset of expression, human marrow and cord blood cells were sorted into three subpopulations, representing stem, multipotent, and erythroid progenitor cells, and the expression of Kell and XK was determined by reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. Mouse Kell and XK transcripts were determined by cDNA blotting of progenitor cells of defined lineage., Results: By RT-PCR, human peripheral blood progenitor cells had weak expression of Kell and XK transcripts but FACS analysis did not detect surface antigens. Kell and XK transcripts are expressed in multipotent progenitor cells and these cells express Kell surface antigens. The expression of Kx antigen in progenitor cells was not determined owing to nonspecific reactions with the antibody. By cDNA blotting, mouse Kell expression was detected in bipotential megakaryocytes-erythroid cells and in colony-forming units-erythroid (CFU-E) and burst-forming units-erythroid (BFU-E), whereas XK was only detected in CFU-E and BFU-E., Conclusion: Both Kell and XK transcripts occur early during erythropoiesis; however, expression may not be coincident because, in mice, Kell transcripts, but not XK, occur in bipotential megakaryocytes-erythroid progenitor cells.

Erythropoietin (Epo) transduces mitogenic and chemoattractant signals to human endothelial cells. Identifications of Epo-responsive genes are important for understanding the molecular nature of Epo signaling in endothelial cells. The effects of Epo on differential expression of various genes were examined in human microvascular endothelial cells (HMVEC) by differential display reverse transcriptase polymerase chain reaction (RT-PCR). In the current study we obtained from Epo-treated HMVEC a cDNA fragment with characteristics of the 3' end of mRNA. Using the cDNA fragment, we then selectively isolated a full-length clone by screening an unamplified endothelial cell cDNA library followed by 5' rapid amplification of cDNA ends by polymerase chain reaction (RACE-PCR). The nucleotide sequence of the longest cDNA revealed an open reading frame of 3311 nucleotides that encodes a protein consisting of approximately 906 amino acids with a predicted MW of approximately 100 kDa. The nucleotide sequence of the cDNA is nearly identical to that of transforming acidic coiled coil-containing (TACC2) and anti-zuai-1 (AZU-1) cDNA clones except at the 5'- and 3'-ends. Northern blot analysis showed an increase in endothelial-TACC-related mRNA levels in Epo-treated cells in comparison to that of the control cells. Endothelial-TACC-related mRNA was highly expressed in heart and skeletal muscle tissue. Placenta and brain tissue exhibited low levels of expression of endothelial-TACC-related gene. Southern blot analysis of genomic DNA from somatic cell hybrids showed that endothelial-TACC-related cDNA maps to chromosome 10. Immunofluorescence microscopy and the occurrence of several putative phosphorylation and SH3 binding sites on the deduced protein suggest that endothelial-TACC-related protein may be involved in Epo signaling cascades in endothelial cells., (Copyright 2001 Academic Press.)