Therapeutic effects of chimeric antigen receptor T cells (CAR-T) on relapse/refractory diffuse large B-cell lymphoma (R/R DLBCL): a meta-analysis.

OBJECTIVE: Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma (NHL). This study aimed to systematically evaluate the efficacy of chimeric antigen receptor T cells (CAR-T) in treating relapse/refractory DLBCL (R/R DLBCL) and associated complete-remission rate (CR). MATERIALS AND METHODS: PubMed, Cochrane Library, CNKI, VIP, CBM, and Wanfang databases were searched, and literature was collected up to January 2019. According to inclusion criteria and exclusion criteria, two researchers independently reviewed and screened literature, extracted required data and crossly checked them. This meta-analysis was conducted using RevMan 5.3 software. RESULTS: This study finally included 13 English literatures and 263 cases. There was no heterogeneity among all these studies, therefore, fixed effect model was used. Meta-analysis findings showed that total CR rate of R/R DLBCL treated with CAR-T was 46.8% (95% CI: 0.408-0.533). Subgroup analysis showed that CR rate of CD28 group was slightly higher [52.5%, with 95% confidence interval (CI): 0.441-0.602] compared to that of 4-1BB group (41.5%, with 95% CI: 0.324-0.510). CR rate of CD19 group was slightly higher (49.2%, with 95% CI: 0.429-0.556) compared to that of CD20 group (42.2%, with 95% CI: 0.231-0.639). Funnel chart of total CR rate, co-stimulatory factor, and target antigen demonstrated fundamental symmetry. Moreover, age, HSCT administration, CAR-T cell counts, and drug pre-treatment also affected immunotherapy on CAR-T on R/R DLBCL. CONCLUSIONS: CAR-T treatment for R/R DLBCL demonstrated evident curative effect and high complete remission rate. CAR-T cell immunotherapy would be expected to become mainstream therapy for hematolymph system tumors.

Phylogenetic Analysis and Fungicide Baseline Sensitivities of Monilia mumecola in China.

Monilia mumecola is one of the causal agents of peach brown rot in China. In this study, M. mumecola isolates from different locations and hosts were used to analyze the genetic diversity and to assay the sensitivity to four generally used fungicides: carbendazim, tebuconazole, azoxystrobin, and boscalid. Results showed that isolates from different locations tended to be separated. Interestingly, isolates from different hosts (e.g., peach and apricot) at the same locations generally clustered together, indicating that the M. mumecola isolates may infect different hosts in the same areas. The fungicide sensitivity assay of 93 M. mumecola isolates showed that the average effective concentration for 50% mycelial growth inhibition values for carbendazim, tebuconazole, azoxystrobin, and boscalid were 0.103, 0.034, 0.325, and 0.419 µg/ml, respectively. The sensitivity distributions of the tested isolates to the four fungicides showed continuous unimodal curves, indicating no qualitative shift of resistance. No significant difference of sensitivity to tested fungicides was observed among isolates from either different locations or different hosts.

Significance of detecting IgH and TCRγ gene rearrangements in patients with hemopoietic maligancies by real-time quantitative PCR.

The aim of our study was to investigate the association of IgH and TCRγ gene rearrangements in hematological malignancies with the disease and clinical application. IgH and TCRγ gene rearrangements were determined in 69 paraffin and bone marrow specimens with SYBR Green I fluorescent dye and RQ-PCR method, including 21 paraffin-embedded tissues of the onset cases and 48 bone marrow samples, representing 15 ALL and 25 AML cases. After chemotherapy, 8 cases were NHL; the 10 cases of the negative control group were healthy people. Among the ALL cases, the IgH rearrangement occurred in 80.0%, the TCRγ rearrangement in 46.7%, and both gene rearrangements in 46.7%. Among the AML cases, the IgH rearrangement occurred in 72.0%, the TCRγ rearrangement in 68.0%, and both gene rearrangements in 60.0%. In the lymphoma cases, the IgH rearrangement occurred in 93.1%, the TCRγ rearrangement in 51.7%, and both gene rearrangements in 44.8%. In the negative control group, the 10 cases were all negative. There was the phenomenon of "sequence-non-fidelity" in the hematologic malignancies; the detection rate of both genes was much higher than that of the single gene. The application of the RQ-PCR method in the detection of IgH and TCRγ gene rearrangements in hematologic malignancies has important clinical significance in MRD monitoring.

Sensitivity of Monilinia fructicola from Peach Farms in China to Four Fungicides and Characterization of Isolates Resistant to Carbendazim and Azoxystrobin.

Brown rot of peach caused by Monilinia fructicola can cause considerable preharvest and postharvest losses in China. Fungicides are increasingly utilized to minimize such losses. Eighty isolates of M. fructicola were collected from commercial peach orchards located in five provinces in China and the sensitivity to carbendazim, azoxystrobin, tebuconazole, and boscalid was determined. Resistance to carbendazim was detected only in the Yunnan province in 15 of 16 isolates. Characterization of carbendazim-resistant isolates revealed stable resistance, no fitness penalty, and negative cross resistance to diethofencarb. Resistant isolates produced disease symptoms on detached fruit sprayed with label rates of formulated carbendazim and possessed the amino acid mutation E198A in ß-tubulin. Resistance to azoxystrobin was detected in 3 of 10 isolates from Fujian. In contrast to carbendazim resistance, however, azoxystrobin resistance was unstable, associated with a fitness penalty, and not associated with mutations in the target gene cytochrome b. The concentration at which mycelial growth is inhibited 50% (EC50) values of the azoxystrobin-sensitive isolates were 0.02 to 1.94 µg/ml, with a mean value of 0.54 µg/ml. All isolates were sensitive to tebuconazole, with a mean EC50 value of 0.03 µg/ml. The EC50 values for boscalid were 0.01 to 3.85 µg/ml, with a mean value of 1.02 µg/ml. Our results indicate that methyl benzimidazole carbamates (MBCs), quionon outside inhibitors, demethylation inhibitor fungicides, and succinate dehydrogenase inhibitors are likely to be very effective in controlling brown rot in many peach production areas in China, but that resistance to MBCs is emerging.

First Report of Brown Rot of Apricot Caused by Monilia mumecola.

In May 2013, apricot (Prunus armeniaca) fruits covered with grayish, conidial masses were collected from an unknown cultivar in an experimental field of Huazhong Agricultural University, Wuhan, Hubei Province. About 3 to 5% of fruit was infected and affected apricots had tan to white zones of sporulation, which resembled brown rot caused by Monilia species. Conidia were harvested from the surface of the sporulating apricot fruit and spread onto shallow potato dextrose agar (PDA) media (about 2 mm in thickness) using sterile cotton swabs. Conidia were lemon-shaped and mean size was 15.7 (10 to 22.5) × 25 (16.25 to 35) µm. Conidia on PDA were incubated at 23°C for 3 h in darkness, then observed under microscope. More than two germ tubes were produced from each conidium, which was the distinctive trait of Monilia mumecola species (2). Single-spore isolates were obtained and 3 isolates were cultured on PDA in petri dishes. Mycelium grew at an average of 15 mm per day, and the colony showed concentric rings of mycelium with lobbed margins at 23°C in darkness. A 712-bp fragment was PCR amplified from ß-tubulin gene (TUB2) of all the nine isolates investigated indicative of M. mumecola (2). The ribosomal ITS1-5.8S-ITS2 regions of nine isolates were also PCR-amplified from genomic DNA using primers ITS1 and ITS4 and then sequenced (4). ITS sequences were identical to ITS sequences of M. mumecola from China (HQ908786) and Japan (AB125613, AB125614, and AB125620), but only has 98% and 97% identity with the closest species M. laxa (EU042149) and M. fructicola (HQ908789) according to BLAST search in GenBank. Pathogenicity was confirmed by inoculating mycelial plugs of three isolates into six surface-sterilized apricots wounded with a 6-mm diameter sterile cork borer. Control fruit received plain PDA plugs and was incubated in a moist chamber at 23°C with 12 h light/12 h dark. All inoculated fruit developed typical brown rot symptoms with sporulating areas as described above after 3 days of incubation, while control fruits remained healthy. The developing spores on inoculated fruit were re-isolated and confirmed to be M. mumecola. M. mumecola was first isolated from Prunus mume in Japan in 1982 as an unknown Monilia species (3), then identified and the nomenclature was provided in 2004 (1). To our knowledge, this is the first report of M. mumecola on P. armeniaca indicating that M. mumecola has spread to different hosts. References: (1) Y. Harada et al. J. Gen. Plant Pathol. 70:297, 2004. (2) M. J. Hu et al. Plos One, 6(9):e24990, 2011. (3) S. Nakao. Kongetsu-no-noyaku, 1:92, 1992. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

First Report of Brown Rot Caused by Monilia mumecola on Chinese Sour Cherry in Chongqing Municipality, China.

Cherry is widely planted in China, from Liaoning, Beijing, Hebei, Shandong, Zhejiang, Jiangsu, and Anhui provinces (eastern China), to Shaanxi, Sichuan, Chongqing, and Guizhou provinces (western China). The brown rot fungus Monilinia fructigena causes considerable production losses in cherry production in Liaoning Province (3). In May 2013, Chinese sour cherry (Prunus pseudocerasus) cv. Wupi displaying symptoms of brown rot was found in an orchard in Chongqing municipality. Diseased cherry fruit had a brown rot sporulating with grayish, conidial tufts. The fruit later succumbed to the soft rot or shivered and became a mummy. Single-spore isolations on PDA resulted in colonies with concentric rings of pigmented mycelium with lobbed margins. Conidia were broadly ellipsoid to subglobose, occasionally even globose, with an average size of 16 × 12.7 µm. Multiple germ tubes were produced from each conidium, a germination pattern unique to Monilia mumecola (1,2,4). The pathogen identity was confirmed by multiplex PCR as described by Hu et al. (2). The PCR resulted in a 712-bp amplicon, which is diagnostic of M. mumecola. Further sequencing of the internal transcribed spacer (ITS) region 1 and 2 and 5.8S gene further indicated 100% identity with that of M. mumecola isolates from China (Accession No. HQ908786) and from Japan (AB125613, AB125614, and AB125620). Koch's postulates were confirmed by inoculating mature cherry fruit with mycelia plugs. Inoculated fruit were placed in a sterilized moist chamber, and incubated at 22°C with 12 h light/dark cycle. Inoculated fruit developed typical brown rot symptoms only 2 days after inoculation, while the control fruit, inoculated with a sterile PDA plug, remained healthy. The pathogen isolated from inoculated symptomatic fruit was confirmed to be M. mumecola based on morphological characteristics and germination pattern. It should be noted that the conidia on inoculated fruit showed an average size of 20 × 15.3 µm, significantly bigger than that of from PDA, and most produced more than three germ tubes. The inoculation experiments were performed in triplicates. M. mumecola was first reported as the causal agent of brown rot of mume in Japan in 2004 (1). Later studies demonstrated that it is also pathogen on other stone fruits, e.g., peach, nectarine (2), and apricot (4). To our knowledge, this is the first report of cherry brown fruit rot caused by M. mumecola, and the first report of M. mumecola in Chongqing municipality. References: (1) Y. Harada et al. J. Gen. Plant Pathol. 70:297, 2004. (2) M. J. Hu et al. Plos One 6(9): e24990, 2011. (3) Z. H. Liu et al. J. Fruit Sci. 29:423, 2012. (4) L. F. Yin et al. Plant Dis. 98:694, 2014.

First Report of Peach Brown Rot Caused by Monilinia fructicola in Central and Western China.

Brown rot of peach (Prunus persica) in China has been reported to be caused by at least three Monilinia species (1). In the present study, peaches with symptoms resembling brown rot caused by Monilinia species were collected from commercial orchards in the northwestern province of Gansu in August 2010, the southwestern province of Yunnan in July 2011, and in the central province of Hubei in July 2012. Affected fruit showed the typical symptoms of brown rot with zones of sporulation. Fungal isolates were single-spored and cultured on potato dextrose agar (PDA). Colonies showed grayness with concentric rings of sporulation after incubation at 25°C in the dark. Mean mycelial growth of isolates YHC11-1a and YHC11-2a from Yunnan, GTC10-1a and GTC10-2a from Gansu, and HWC12-14a and HWC12-23a from Hubei, was 4.6 ± 0.4 and 7.5 ± 0.7 cm after 3 and 5 days incubation, respectively. Conidia were lemon shaped and formed in branched monilioid chains, and the mean size was 9.3 (6.7 to 11.5) × 12.5 (7.9 to 17.8) µm, which was consistent with the characteristics of Monilinia fructicola (1,2). The species identification was confirmed by sequencing of the ribosomal ITS sequences. The ribosomal ITS1-5.8S-ITS2 region was amplified from each of the six isolates using primers ITS1 and ITS4 (3). Results indicated that the ITS sequences of these isolates were identical and showed the highest similarity (100%) with M. fructicola ITS sequences from isolates collected in China (GenBank Accession Nos. HQ893748, FJ515894, and AM887528), Slovenia (GU967379), Italy (FJ411109), and Spain (EF207423). The pathogen was also confirmed to be M. fructicola based on the detection of an M. fructicola- specific band (534 bp) using a PCR-based molecular tool developed for distinguishing Chinese Monilinia species affecting peach (1). Pathogenicity was tested on surface-sterilized, mature peaches (Shui Mi Tao) with representative isolates. Fruits were holed at three equidistant positions to a depth of 5 mm using a sterile cork borer. Mycelial plugs (5 mm in diameter) from the periphery of a 4-day-old colony of each isolate were placed upside down into each hole, control fruits received water agar. After 3 days of incubation at 22°C in a moist chamber, inoculated fruits developed typical brown rot symptoms while control fruits remained healthy. Pathogens from the inoculated fruit were confirmed to be M. fructicola based on morphological characteristics. To our knowledge, this is the first report of occurrence of M. fructicola in Gansu, Yunnan, and Hubei provinces, thousands of kilometers away from eastern China where occurrence of peach brown rot caused by M. fructicola has been confirmed (2,4). The results indicated the further geographical spread of the M. fructicola in China. References: (1) M. J. Hu et al. Plos One 6(9):e24990, 2011. (2) M. J. Hu et al. Plant Dis. 95:225, 2011. (3) T. J. White et al. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Academic Press, San Diego, 1990. (4) X. Q. Zhu et al. Plant Pathol. 54:575, 2005.

Tuning the metal-insulator transition in manganite films through surface exchange coupling with magnetic nanodots.

In strongly correlated electronic systems, the global transport behavior depends sensitively on spin ordering. We show that spin ordering in manganites can be controlled by depositing isolated ferromagnetic nanodots at the surface. The exchange field at the interface is tunable with nanodot density and makes it possible to overcome dimensionality and strain effects in frustrated systems to greatly increasing the metal-insulator transition and magnetoresistance. These findings indicate that electronic phase separation can be controlled by the presence of magnetic nanodots.

First Report of Brown Rot of Peach Caused by Monilinia fructicola in Southeastern China.

In 2009 and 2010, peaches (Prunus persica) with brown rot symptoms were collected from Zhejiang Plant Protection State Research Farm and a commercial orchard in Fujian Province in southeastern China. Affected fruit showed brown decay with zones of sporulation. Single-spore isolates from the diseased fruit were cultured on potato dextrose agar. After incubation at 25°C in the dark for 5 days, colonies were gray with concentric rings of sporulation. Mean mycelial growth of isolates MZ09-2a from Zhejiang Province and 0907-a from Fujian Province was 4.46 ± 0.58 and 7.05 ± 0.81 cm after 4 and 7 days of incubation, respectively. Lemon-shaped conidia were formed in branched, monilioid chains and mean size was 14.6 (9.6 to 21.6) × 10.3 (7.2 to 13.2) µm. Mean conidial germination was 97 ± 1% with one straight germ tube per conidium. These characteristics were consistent with descriptions of Monilinia fructicola (G. Wint.) Honey (3). Morphology-based species identification was confirmed by sequencing and analysis of ribosomal internal transcribed spacer (ITS) sequences. A 496-bp fragment including ITS 1 and 2 and the gene encoding the 5.8S small subunit of the ribosomal RNA from isolates MZ09-2a and 0907-a was amplified using the universal primer pair ITS1/ITS4 (4) and sequenced. Nucleotide sequences of both isolates were identical. Blast searches of the ITS sequences in GenBank showed the highest similarity (100%) with sequences of M. fructicola isolates from China (FJ515894), Italy (FJ411109), and Spain (EF207423). The isolates were also identified as M. fructicola using the Monilinia spp. PCR detection protocol based on sequence-characterized amplification region marker DNA sequences (2). Pathogenicity was confirmed by inoculating surface-sterilized, mature cv. Zhonghua 2 peaches with mycelial plugs of representative isolates. Fruit was stabbed at two points with a 5-mm-diameter sterile cork borer, mycelial plugs (5 mm in diameter) were removed from the periphery of a 4-day-old colony of each isolate and placed upside down into each wound; control fruit received water agar. Inoculated fruit developed typical brown rot symptoms with sporulating fungi while control fruit remained healthy after 3 days of incubation at 22°C in a moist chamber. Pathogens were reisolated from the inoculated fruit and confirmed to be M. fructicola on the basis of morphological characteristics. To our knowledge, this is the first report of M. fructicola in Zhejiang and Fujian provinces. Both provinces are located more than 1,000 km south of Beijing, Hebei, and Shandong provinces, where M. fructicola had been reported previously (1). References: (1) J. Y. Fan et al. Acta Phytophylacica Sin. (in Chinese) 34:289, 2007. (2) I. Gell et al. J. Appl. Microbiol. 103:2629, 2007. (3) G. C. M. van Leeuwen and H. A. van Kesteren. Can. J. Bot. 76:2041, 1998. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds., Academic Press, San Diego, 1990.

Tunable metallicity of the La5/8Ca3/8MnO3(001) surface by an oxygen overlayer.

We studied the surface structure of La_{5/8}Ca_{3/8}MnO_{3}(001) thin films using in situ scanning tunneling microscopy (STM). Atomically resolved STM images reveal that a (sqrt[2]xsqrt[2])R45;{ degrees } reconstructed surface and a (1x1) surface can be converted back and forth through adsorption and desorption of oxygen at the surface. The electrical properties of the surfaces are investigated by scanning tunneling spectroscopy. I-V curves clearly show that the presence of an oxygen overlayer renders the surface insulating while the (1x1) surface without the oxygen overlayer is metallic.

Time-resolved electronic phase transitions in manganites.

The dynamics of first-order electronic phase transitions in complex transition metal oxides are not well understood but are crucial in understanding the emergent phenomena of electronic phase separation. We show that a manganite system reduced to the scale of its inherent electronic charge-ordered insulating and ferromagnetic metal phase domains allows for the direct observation of single electronic phase domain fluctuations within a critical regime of temperature and magnetic field at the metal-insulator transition.

Reemergent metal-insulator transitions in manganites exposed with spatial confinement.

The metal-insulator transition is characterized as a single peak in the temperature-dependent resistivity measurements; exceptions to this have never been seen in any single crystal material system. We show that by reducing a single crystal manganite thin film to a wire with a width comparable to the mesoscopic phase-separated domains inherent in the material, a second and robust metal-insulator transition peak appears in the resistivity versus temperature measurement. This new observation suggests that spatial confinement is a promising route for the discovery of emergent physical phenomena in complex oxides.

Magnetocrystalline anisotropy in permalloy revisited.

Permalloy with a body-centered-cubic structure has been grown on GaAs(001) by molecular beam epitaxy. Its magnetism, Curie temperature, and magnetic anisotropy are determined experimentally and compared to those of conventional face-centered-cubic Permalloy. Unexpectedly the vanishing magnetic cubic anisotropy in Permalloy is found to be independent of its atomic structure but depends only upon the stoichiometry of Fe and Ni in the FexNi1-x alloy. This observation is further investigated and confirmed by first-principles electronic band calculations, which help to understand the long-standing issue of why Permalloy should be a soft magnet.

Body-centered-cubic Ni and its magnetic properties.

The body-centered-cubic (bcc) phase of Ni, which does not exist in nature, has been achieved as a thin film on GaAs(001) at 170 K via molecular beam epitaxy. The bcc Ni is ferromagnetic with a Curie temperature of 456 K and possesses a magnetic moment of 0.52+/-0.08 micro(B)/atom. The cubic magnetocrystalline anisotropy of bcc Ni is determined to be +4.0x10(5) ergs x cm(-3), as opposed to -5.7x10(4) ergs x cm(-3) for the naturally occurring face-centered-cubic (fcc) Ni. This sharp contrast in the magnetic anisotropy is attributed to the different electronic band structures between bcc Ni and fcc Ni, which are determined using angle-resolved photoemission with synchrotron radiation.