Construction and Identification of Leukemia Cell Line Stably Expressing CD123 and CLL-1 / 中国实验血液学杂志

OBJECTIVE@#To construct an acute myeloid leukemia cell line stably expressing CD123-CLL1 so as to provide an "in vitro" model for studying the role of CD123 and CLL-1 in leukemia and the treatment targeting CD123 and CLL-1.@*METHODS@#The recombinant plasmid of lentivirus was constructed by synthesizing CD123 and CLL-1 sequences and PCR homologous recombination. The lentivirus vector was packaged by three-plasmid packaging system. After collecting the supernatant of lentivirus, the virus titer was determined by quantitative PCR. K562 leukemia cells were collected and transtected with virus supernatant. Leukemia cell line stably expressing the target gene were screened by purinomycin. The expression levels of CD123 and CLL-1 were detected by RT-PCR and flow cytometry.@*RESULTS@#The lentiviral vector was successfully constructed, and identified by agarose gel electrophoresis and gene sequencing, then the virus titer of the supernatant was up to 5.81×10@*CONCLUSION@#Lentiviral vector expressing CD123-CLL1 has been successfully constructed, and K562 leukemia cell line stably expressing CD123 and CLL-1 has been successfully obtained.

[Simulation and validation of diffuse radiation in Qianyanzhou area, Jiangxi, China].

Accurate estimation of diffuse radiation is of great significance for evaluating its effect on terrestrial ecosystem carbon exchange. Based on the observed diffuse radiation data in the meteorological observation field in mid-subtropical Qianyanzhou, Jiangxi, China from March 1, 2012 to February 28, 2013, the simulated results of five widely used diffuse radiation decomposition models (Reindl-1, Reindl-2, Reindl-3, Boland, BRL) were validated. The results indicated that, on the 30 min scale, all of the five models could well simulate the diffuse radiation of this area overall. But the effect of models decreased significantly with the rising of clearness index (kt). Especially when kt>0.75, each model was.unable to simulate diffuse radiation in the region. Regarding the simulation of seasonal change of diffuse radiation, the five models could simulate diffuse radiation well in most months. Relative deviation between simulated and observed values of yearly diffuse radiation of five models had a maximum of 7.1% (BRL), a minimum of 0.04% (Reindl-1), and an average of 3.6%. The simulated values of the five models appeared to be overestimated in the summer when radiation was strongest, temperature was highest, and precipitation was relatively low. For example, in July, the diffuse radiation was overestimated by 14.5%-28.2%, 21.2% on average. This was primarily due to the. method of estimating diffuse radiation under the condition of high kt. The uncertainty requires further evaluation in the model application. Considering the results of validation, simulation precision and the accessibility of input variables, the order of the simulation performance of five models was BRL>Reindl-3>Reindl-2>Reindl-1 >Boland.

[Comparative observation of diffuse radiation in Qianyanzhou during the spring of 2012].

Global radiation and diffuse radiation were measured from March to June of 2012 in Qianyanzhou Experimental Station of Red Soil and Hilly Land, Chinese Academy of Sciences by ising three types of pyranometers, including CMP11 attached with a shadow ring, SPN1 and RSR3, which were placed in parallel. The observations showed that both global radiation and diffuse radiation measured by these pyranometers had a good linear correlation. The global radiation measured by SPN1 and RSR3 was respectively 3. 0% and 20.5% lower than that measured by the CMP 11. The diffuse radiation measured by SPN1 and RSR3. was respectively 5.5% and 7.9% lower of than that measured by the CMP11. Under the sunny, cloudy and overcast conditions, the daily variations of diffuse radiation measured by the three pyranometers were similar, and hence, the diffuse radiation values at a specific solar elevation angle measured by the different pyranometers were also similar. There was difference in daily accumulative diffuse radiation measured by the different pyranometers. Monthly diffuse fractions of March, April and May were 0.56, 0.59 and 0.70, respectively. In the subtropical area of southern China, the diffuse radiation accounted for a relatively large proportion of the global solar radiation and varied considerably over time. It is necessary to conduct long-term continuous measurements to capture the variability of diffuse radiation over different underlying surfaces.

Allogeneic CAR-T for treatment of relapsed and/or refractory multiple myeloma: four cases report and literatures review / 中华血液学杂志

Objective: To investigate the safety and efficacy of allogeneic CAR-T cells in the treatment of relapsed/refractory multiple myeloma (RRMM) . Methods: CAR-T cells were prepared from peripheral blood lymphocytes of HLA mismatch healthy donors. Median age was 55 (48-60) . Allogeneic cells were derived from 3 HLA haploidentical donors and 1 HLA completely mismatch unrelated donor. Four patients with RRMM were conditioned with FC regimen followed by CAR-T cell transfusion. They were infused into CART-19 (1×10(7)/kg on day 0) and (4.0-6.8) ×10(7)/kg CART-BCMA cells as split-dose infusions (40% on day 1 and 60% on day 2) . The adverse reactions and clinical efficacy were observed during follow-up after infusion, and the amplification and duration of CAR-T cells in vivo were monitored by PCR technique. Results: CAR-T cells were successfully infused in 3 of the 4 RRMM patients according to the study plan, and the infusion in one patient was delayed by 1 day due to high fever and elevated creatinine levels on day 3. The side effects included hematological and non-hematological toxicity, grade 3 hematological toxicity in 2 patients, grade 3 CRS in 1 one, grade 1 CRES in 1 one, prolonged APTT in 3 ones, tumor lysis syndrome in 1 one, mixed chimerism detected STR and clinical GVHD manifestation in 1 one. According to the efficacy criterias of IMWG, 2 patients acquired PR, 1 MR, and 1 SD respectively. Progression-free survival was 4 (3-5) weeks and overall survival was 63 (3-81) weeks. CAR T cells were amplified 2.2 (2-14) times in the patients with a median survival time of 10 (8-36) days. Conclusions: Small sample studies suggested that GVHD may be present in the treatment of RRMM with allogeneic CAR-T cells. There were early clinical transient events after transfusion. Low amplification and short duration of CAR-T cells in vivo may be the main factors affecting the efficacy.

Diagnosis of intravenous leiomyomatosis extending to heart with emphasis on magnetic resonance imaging / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Intravenous leiomyomatosis is a rare neoplasm, and its cardiac extension is often overlooked or misdiagnosed. The purpose of this study was to explore the imaging features of intravenous leiomyomatosis with cardiac extension, especially the value of magnetic resonance imaging in differential diagnosis.</p><p><b>METHODS</b>Between July 2005 and August 2008, 4 cases of intravenous leiomyomatosis with cardiac extension were resected in Cangzhou Central Hospital. Three cases had echocardiography performed, two had post contrast scans of CT performed, and two had MRI performed. Between July 2005 and May 2010, 25 cases of histopathologically proven other kinds of tumors involving the inferior vena cava and right atrium were compared for discussion of differential diagnosis.</p><p><b>RESULTS</b>Intravenous leiomyomatosis with cardiac extension demonstrated a hyperechoic elongated mobile mass extending from the inferior vena cava to the right atrium with or without evidence of protruding into the right ventricle on echocardiography. The lesion was enhanced heterogeneously on post contrast scans of CT and was of relatively lower density compared to the enhanced blood in the inferior vena cava and right atrium, with common iliac vein and the ipsilateral internal iliac and ovarian veins involved in some cases. The untreated uterus myoma demonstrated enlargement of the uterus with heterogeneous contrast enhancement. On MRI, the lesion looked like a luffa vegetable sponge on FIESTA coronal images and a sieve pore on T2-weighted axial images. All four tumors were removed successfully, and follow up of one to four years revealed no recurrence. The 25 cases of histopathologically proven other kinds of tumors involving inferior vena cava and right atrium had their own imaging features different from those seen on intravenous leiomyomatosis with cardiac extension. With reference to their medical history, differential diagnosis can often be made.</p><p><b>CONCLUSION</b>The imaging appearance of intravenous leiomyomatosis has some unique features, and the luffa vegetable sponge and sieve pore like appearance on MRI may be helpful for differential diagnosis.</p>

Preliminary study on CT perfusion imaging in guiding biopsy of pulmonary lumps / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>CT perfusion imaging (CTP) has been proved to be a powerful functional imaging technique. This study aimed to evaluate the value of CTP in guiding biopsy of pulmonary lumps.</p><p><b>METHODS</b>A total of 147 patients with pulmonary lumps who had CT guided biopsies were enrolled in this study from February 2005 to June 2007. The patients were assigned to 3 groups: 33 cases guided by CTP as group I, 45 cases guided by contrast-enhanced scan of CT as group II, and 69 cases guided by plain scan of CT as group III. Each group was subdivided into central and peripheral types according to the location of the lumps. The achievement ratio of biopsy, the accuracy in grouping, and grading of lung cancer, and the incidence of complication were compared.</p><p><b>RESULTS</b>The total achievement ratios of biopsy from group I to III were 100% (33/33), 91% (41/45), and 80% (55/69) respectively, and the difference was statistically significant between group I and III (P < 0.05). For the central type, they were 100% (18/18), 88% (15/17), and 79% (11/14) respectively, and the difference was also statistically significant between group I and III (P < 0.05). For the peripheral type, they were 100% (15/15), 93% (26/28), and 80% (44/55) respectivelies, and the difference was not statistically significant among the three groups. The total accuracies in grouping and grading of lung cancer from group I to III were 100% (27/27), 91% (31/34), and 72% (33/46) respectively, and the difference was statistically significant between group I and III and between group II and III (P < 0.05). For the central type, they were 100% (16/16), 94% (16/17), and 70% (8/12) respectively, and the difference was statistically significant between group I and III (P < 0.05). For the peripheral type, they were 100% (11/11), 88% (15/17), and 72% (26/36) respectively, and the difference was statistically significant between group I and III (P < 0.05). The total incidence of complication from group I to III were 15% (5/33), 27% (12/45), and 43% (30/69) respectively, and the difference was statistically significant between group I and III (P < 0.01). For the central type, they were 11% (2/18), 24% (4/17), and 57% (8/14) respectively, and the difference was statistically significant between group I and III (P < 0.01). For the peripheral type, they were 20% (3/15), 29% (8/28), and 40% (22/55) respectively, and no statistically significant difference was found among the three groups.</p><p><b>CONCLUSIONS</b>CTP guided biopsy of pulmonary lumps using multi-detector row CT has the potential to improve the accuracy of histopathological diagnosis with a lower risk and higher achievement ratio. More research and technical improvements are needed before it is widely used.</p>