Ag-carbon dots with peroxidase-like activity for colorimetric and SERS dual mode detection of glucose and glutathione.

Currently, nanozymes have made important research progress in the fields of catalysis, biosensing and tumor therapy, but most of nanozymes sensing systems are single-mode detection, which are easily affected by environment and operation, so it is crucial to construct nanozymes sensing system with dual-signal detection to obtain a more stable and reliable performance. In this paper, Ag-carbon dots (Ag-CDs) bifunctional nanomaterials were synthesized using carbon dots as reducing agent and protective agent by a facile and green one-step method. A simple and sensitive colorimetric-SERS dual-mode sensing platform was constructed for the detection of glucose and glutathione(GSH) in body fluids by taking advantage of good peroxidase-like and SERS activities of Ag-CDs. Ag-CDs catalyzes H2O2 to hydroxyl radicals(•OH), which oxidized TMB to form ox-TMB blue solution with characteristic absorption peak at 652 nm and Raman characteristic peak at 1607 cm-1. Ag-CDs sensing method exhibited high performance for glucose and GSH with detection limits for colorimetric and SERS as low as 11.30 µM and 3.54 µM, 0.38 µM and 0.24 µM respectively (S/N = 3). In addition, Ag-CDs have good stability and uniformity, ensuring long-term applicability of catalytic system. This colorimetric-SERS dual-mode sensing platform can be used for the determination of glucose and GSH in saliva and urine, and has the advantages of simple, low cost, rapid, and high accuracy, which has a potential application prospect in biosensor and medical research.

Evolving of treatment paradigms and challenges in acute promyelocytic leukaemia: A real-world analysis of 1105 patients over the last three decades.

Although acute promyelocytic leukaemia (APL) is a highly curable disease, challenges of early death (ED) and relapse still exist, and real-world data are scarce in the ATRA plus ATO era. A total of 1105 APL patients from 1990 to 2020 were enrolled and categorized into three treatment periods, namely ATRA, ATRA plus ATO, and risk-adapted therapy. The early death (ED) rate was 20.2%, 10.1%, and 7.0%, respectively, in three periods, while there was no significant decline in the 7-day death rate. Consistently, the overall survival (OS) and disease-free survival (DFS) of APL patients markedly improved over time. Despite the last two periods exhibiting similar DFS, the chemotherapy load was substantially lower in Period 3. Notably, leveraging older age and higher WBC count (especially > 50 × 109/L), we could identify a small group of extremely high-risk patients who had a very high ED rate and poor prognosis, while those with NRAS mutations and higher WBC tended to relapse, both representing obstacles to curing all patients. In conclusion, the evolvement of treatment paradigms can reduce the ED rate, improve clinical outcomes, and spare patients the toxicity of chemotherapy. Special care and innovative agents are warranted for the particularly high-risk APL.

[Advance of a representative traditional Tibetan medicine Meconopsis horridula on its phytochemical and pharmacological aspects].

Meconopsis horridula is one of alpine plants belonging to family Papaveraceae, mainly distributed in Himalaya Range area. M. horridula is a rare alpine flower, and is a kind of traditional Tibetan medicine, which has been included in more than 40 compound formulae, having efficacies of clearing away heat and alleviating pain, activating blood circulation to remove stasis, traditionally used for the treatment of fractures, injuries, and chest and back pains. Modern research shows that the whole plant of M. horridula contains alkaloids, flavonoids, and terpenes, and its pharmacological activities including antitumor, antivirus and myocardial protection etc. However, due to various factors, the current research of M.horridula still faces many challenges. This paper summaries herein a progress of MH on its ecological resources, traditional uses, and studies on chemical constituents and pharmacological effects, hopefully to provide a useful reference for the ecological protection and applications.

[A phytochemical and pharmacological advance on Ilex asprella].

Ilex asprella is one of representative medicinal plants in South of the Five Ridges of China. The roots and rhizomes of I. asprella have the effects of clearing heat and detoxifying, stimulating salvia, and reducing thirst, which has been used to treat wind-heat cold, acute and chronic pharyngitis, and sore throat. Contemporary studies showed that I. asprella contains the major triterpenoids and glycosides, phenolic acids, and minor steroids. The extracts and compounds show activities of anti-inflammatory, antiviral, anti-tumor, and regulating lipid metabolism.The present paper summarizes a phytochemical and pharmacological advance on this species to provide reference for clarification of its pharmacologically active ingredients, quality evaluation, and further explorations.

Thrombopoietic stimulating activity of rhTyrRS (Y341A).

Tumor radiotherapy induces hematopoietic organ damage and reduces thrombocyte counts. Thrombocytopenia is a common disease. Some studies have shown that tRNA synthetase plays not only catalytic tRNA aminoacylation roles, but also functions similarly to cytokines. Recombinant human tyrosyl-tRNA synthetase with a mutated Y341A (rhTyrRS (Y341A)) promotes megakaryocyte migrate from bone marrow to peripheral blood. It would promote megakaryocytes in the lungs adhering to vascular endothelial cells and resulting in the platelet production. The purpose of this research was to investigate the efficacy of rhTyrRS (Y341A) as a therapy for thrombocytopenia and to explore its mechanism of action. We found platelet number was effectively increased by rhTyrRS (Y341A) via platelet count and reticulated platelets (RPs) flow cytometry. We also demonstrated radiation-induced thrombocytopenia could be prevented by rhTyrRS (Y341A). The results of immunohistochemistry and H&E staining showed the number of pulmonary mature megakaryocytes was significantly increased in rhTyrRS (Y341A) treated groups. In transgenic zebrafish larvae, confocal microscopy results showed rhTyrRS (Y341A) promoted the migration and adhesion of megakaryocytes. These results suggested that rhTyrRS (Y341A) promote megakaryocytes in bone marrow migrating to lungs through blood circulation. rhTyrRS (Y341A) may be an effective medicine which could be used to treat patients suffering from thrombocytopenia.

Characterisation and immunomodulating activities of exo-polysaccharides from submerged cultivation of Hypsizigus marmoreus.

Exo-polysaccharides were purified and characterized from the fermentation broth of Hypsizigus marmoreus, a popular edible mushroom consumed in Asia. Among them, B-I-I and B-II-I exhibited potent complement fixating activity, meanwhile, B-N-I, B-I-I, B-II-I and B-II-II exhibited significant macrophage stimulating activity. Molecular weights of the four exo-polysaccharides were determined to be 6.3, 120, 150 and 11 kDa respectively. Molecular characterisation showed that B-N-I is basically an α-1→4 glucan, with branches on C6; B-I-I is a heavily branched α-mannan with 1→2 linked main chain. B-II-I and B-II-II, have a backbone of rhamno-galacturonan with 1→2 linked l-rhamnose interspersed with 1→4 linked galacturonic acid. Structure-activity relationship analysis indicated that monosaccharide compositions, molecular weight, certain structural units (rhamno-galacturonan type I and arabinogalactan type II) are the principal factors responsible for potent complement fixating and macrophage-stimulating activities. Their immunomodulating activities may, at least partly, explain the health benefits of the mushroom.

Efficient purification of antiproliferative polysaccharides from Hypsizigus marmoreus with radial flow chromatography.

The increasing commercial significance of natural polysaccharides for use in medicinal products is stimulating the development of efficient and easy scale-up techniques for polysaccharide purification. In this research, the crude polysaccharides from submerged cultivation broth of Hypsizigus marmoreus were purified using radial flow chromatography (RFC), and the antiproliferative activity of the purified fractions was evaluated in vitro. DEAE Sepharose CL-6B was selected to be packed in the RFC column based on its good resolution, physical stability, and low cost. Compared with axial flow chromatography (AFC), an efficient chromatographic process with significantly less time and buffer consumption but yielding higher polysaccharide recovery and resolution was established in RFC, which could clearly purify the crude polysaccharides into different fractions. An acceptable linear scale-up effect of RFC from 100 to 500 mL was successfully achieved without loss of resolution and enhancement of time consumption. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays in cell cultures indicated that the purified polysaccharide fractions possess moderate antiproliferative activities in three different human cancer cell lines, but have significantly lower cytotoxicity in normal human cell lines in vitro. Among the polysaccharide fractions, the main purified acidic fraction W-I could be considered as a novel potential antitumor agent candidate for several tumors, especially for human alveolar epithelial tumors. This research confirmed for the first time that RFC would be a new fast and efficient tool for purification of polysaccharides into different fractions, both at laboratory and commercial scales.

[Comparison of chromosome karyotype between myelodysplastic syndrome and acute leukemia patients confirmed at the same period].

This study was purposed to compare and analyze the relationship between the abnormality of chromosome karyotypes and diagnosis, prognosis of MDS and AML patients, as well as to explore the characteristics of chromosome prognostic stratification in MDS and AML patients of different ages. The cytogenetic karyotype analysis was performed in 134 cases of MDS and 123 cases of AML by using bone marrow short-term culture and R-banding technique. The results indicated that the detected rates of chromosome abnormal karyotypes in MDS and AML patients were 41% and 61% respectively. The abnormal karyotype analysis of MDS and AML group showed that the abnormal karyotype in MDS group displayed number abnormality as the dominate (mainly the +8), while the abnormal karyotype in AML group displayed structure abnormality as the dominant [mainly, t(15;17) and t(8;21)]. The detected abnormal karyotype are mainly for the +8 which has ambiguous correlation with FAB subtype; the detection rates of complex karyotype abnormalities, favourable prognosis karyotype as well as poor prognosis karyotype in the MDS group obviously higher than that of AML group. Among patients with MDS transformed into AML, 12 cases had chromosome abnormal karyotype. There were 3 cases of chromosome abnormal karyotype in AML group which were transformed by MDS. The analysis of age stratification between two groups showed that the detected rate of abnormal karyotype was enhanced with the increase of age in MDS group, and detected rate in ≥ 60 years old group was obviously higher than that in patients with ≤ 30 age group.The detected rate of complex karyotype abnormalities in three age groups of MDS did not show statistical difference; the detected rate of abnormal karyotype in AML group decreased with the increase of age, the detected rate in ≤ 30 years old group was obviously higher than that in ≥ 60 age group,while the detection rate of complex karyotype abnormalities showed that the detected rate in patients ≥ 60 years old group was obviously higher than that in patients with ≤ 30 years old group; Analysis of karyotype prognosis revealed that the detected rate of poor prognosis karyotype increased along with the age growth both in MDS and AML groups, and detected rate in ≥ 60 years old group was obviously higher than that in ≤ 30 years old group; while analysis of favourable prognosis karyotype in MDS and AML group showed that the detected rate in ≤ 30 years old group was obviously higher than that in ≥ 60 years old group. It is concluded that the patients with MDS and AML have higher chromosomal abnormalities,which have important reference value for the diagnosis, treatment and prognosis, meanwhile, the analysis of chromosome karyotype provides an important basis for prognostic stratification.

Polysaccharides with immunomodulating properties from the bark of Parkia biglobosa.

The bark of Parkia biglobosa is used in traditional medicine to cure a wide range of illnesses. Polysaccharides were extracted from the bark with 50% ethanol-water, 50°C and 100°C water, and seven active fractions obtained by anion exchange chromatography and gel filtration. The complement fixation and macrophage stimulating activities of the different fractions were determined. The acidic fractions PBEII-I and PBEII-IV were the most active in the complement fixation assay, but the other fractions were also potent compared to the positive control BPII from Biophytum petersianum. Fractions PBEII-I and PBEII-IV were also the most potent fractions in stimulating macrophages to release nitric oxide. Structural studies showed that PBEII-I and PBEII-IV were pectic type polysaccharides, containing arabinogalactan type II structures. The observed differences in biological activities among the seven purified polysaccharide sub-fractions are probably due to differences in monosaccharide compositions, linkage types and molecular sizes.

A comparison of bioactive aqueous extracts and polysaccharide fractions from roots of wild and cultivated Cochlospermum tinctorium A. Rich.

In Malian traditional medicine the roots of Cochlospermum tinctorium are used in the treatment of gastric ulcer, but extending harvesting is causing a growing concern of a dramatic reduction in the wild plant population. In the present study cultivation of C. tinctorium is evaluated, and structural components and bioactive properties of crude water extracts and isolated polysaccharide fractions from roots of wild and cultivated C. tinctorium are compared. The crude water extracts were shown to contain starch, pectin- and inulin-type polysaccharides, in addition to phenolic substances and protein, while the isolated acidic polysaccharide fractions contained mainly monosaccharides typical for pectins. The monosaccharide compositions of the polysaccharide fractions from roots of wild versus cultivated plants were comparable, albeit the yields in the cultivated roots were lower. Furthermore, the crude extracts and isolated polysaccharide fractions from wild and cultivated roots exhibited similar complement fixating activities, but were not able to activate macrophages. The crude extracts from cultivated roots were also shown to be moderate radical scavengers. The present study has shown that roots of cultivated C. tinctorium contain the same types of bioactive polysaccharides as the wild roots. However, in order to utilize roots of cultivated C. tinctorium in traditional medicine the cultivation method should be improved.

Effect of mechanical stretch on the expressions of elastin, LOX and Fibulin-5 in rat BMSCs with ligament fibroblasts co-culture.

The occurrence of PFD is closely related with elasticity, toughness, and functional changes of the connective tissue of the pelvic support tissue. This study aims to evaluate the effect of mechanical stretch on the differentiation of BMSCs with a co-culture with pelvic ligament fibroblasts. BMSCs was isolated and identified from 7 day SPF SD rats. Rat pelvic ligament fibroblasts were obtained from rat pelvic ligament. The fourth passage of fibroblasts was subjected to 10% deformation with 1 Hz, 12 h periodic one-way mechanical stretch stimulation, and the cells were then co-cultured with BMSCs. The longer co-culture and co-culture with mechanical stretch (i.e. 6 and 12 days) induced more expression of elastin, LOX, and Fibulin-5, compared to the groups without stimulation. Compared to co-culture group each, Co-culture with mechanical stretch stimulation group induced significant expression of elastin, LOX, and Fibulin-5, both in 3, 6 and 12 days co-culture groups (P < 0.05). However, there were no significant differences among 3, 6, and 12 days control groups. These results suggest that in an indirect co-culture system, pelvic ligament fibroblasts with mechanical stretch stimulation can promote BMSCs differentiation, reflecting in the increased expression of elastin, LOX, and Fibulin-5.

[Comparison of immunological reconstitution and related complications after HLA-matched and HLA haploidentical peripheral blood hematopoietic stem cell transplantation].

OBJECTIVE: To explore the difference of immune function and relationship with main complications after HLA-matched and HLA haploidentical allogeneic peripheral blood hematopoietic stem cell transplantation (allo-PBSCT). METHODS: Sixty-seven patients undergoing HLA-matched (n = 33) or HLA haploidentical (n = 34) allo-PBSCT during the same time period in our hospital from June 2004 to December 2007 were included in this study. Indirect immunofluorescence assay was employed to detect lymphocyte subsets before transplantation and on month 1, 3, 6, 12 and 18 after transplantation and the lymphocyte subsets of 100 healthy people were used as normal control. The comparison of immunological reconstitution and relationship with main complications was carried out with statistical analysis. RESULTS: (1) Comparison of the 67 patients with normal controls showed that CD(3)(+), CD(4)(+), CD(4)(+)/CD(8)(+) at month 1, CD(4)(+), CD(4)(+)/CD(8)(+) at month 3 and CD(4)(+) at month 6 after PBSCT were lower. CD(8)(+) at month 3 and month 6 were higher. (2) The immune function was not statistically different between HLA haploidentical and HLA-matched allo-PBSCT (P > 0.05). (3) The immune function of patients with and without severe infection was not statistically different (P > 0.05). (4) The immune function of patients with chronic graft-versus-host disease (cGVHD) between HLA haploidentical and HLA-matched allo-PBSCT groups was not statistically different. The immune function of patients without cGVHD in two groups was not statistically different (P > 0.05). (5) The immune function of patients with or without relapse was not statistically different (P > 0.05). CONCLUSIONS: HLA-haploidentical PBSCT conditioning including antithymocyte globulin without in vitro T cell depletion is feasible and safe. The immunological reconstitution, incidence of severe infection, incidence of relapse and treatment-related mortality are not significantly different between HLA-matched and HLA haploidentical allo-PBSCT.

[Non-T-cell depleted HLA haploidentical peripheral blood stem cell transplantation for hematological malignancies: report of 36 cases].

OBJECTIVE: To analyze the clinical outcome of human leukocyte antigen (HLA) haploidentical peripheral blood stem cell transplantation (PBSCT) from related donors for hematological malignancies. METHODS: Thirty-six patients with hematological malignancies, with a median age of 25 (11-48) years, were transplanted with PBSC from an HLA-haploidentical family donors: 7 were 1 locus mismatched and 29 were 2-3 loci mismatched. The recipients received myeloablative conditioning regimen, in combination with different immunosuppressants according to the degree of HLA disparity followed by non-T-cell depleted PBSCT. The median number of CD34+ cells were 11 (4.16-21.00) x 10(6)/kg. RESULTS: All patients achieved sustained, full donor-type engraftment. Fifteen patients (41.7%) developed grade I-II aGVHD. Among 29 patients followed up more than 18 months, 17 (58.6%) developed cGVHD. There was no statistical difference in decrease and recovery of T, B and NK cell subsets after transplantation between HLA haploidentical group and HLA identical PBSCT group. The median follow-up duration was 15 (4 -69) months. Five patients (13.9% ) relapsed. The 2-year probability of leukemia-free survival (LFS) was 82.2%. CONCLUSION: Non-T-cell depleted HLA-haploidentical PBSCT is safe and feasible for patients with hematological malignancies after myeloablative conditioning regimen combined with intensive immunosuppressants.

[The value of multiplex fluorescence in situ hybridization in the detection of complex karyotypic abnormalities of acute myeloid leukemia].

OBJECTIVE: To investigate the value of multiplex fluorescence in situ hybridization (FISH) in the detection of complex karyotypic abnormalities of acute myeloid leukemia (AML). METHODS: Multiplex FISH was used in combination with conventional cytogenetics (CC) and interphase FISH to study 14 cases of AML with complex karyotypic abnormalities. RESULTS: In the 14 cases of AML studied, conventional cytogenetics detected 23 numerical and 56 structural chromosome abnormalities. Among them 4 gained whole chromosome and 4 lost whole chromosome which were confirmed by multiplex FISH. Twelve chromosome losses detected by CC were revised as derivative chromosomes resulted from various structural aberrations, and 26 derivative and 19 marker chromosomes were characterized precisely by multiplex FISH. Most of them were resulted from unbalanced translocations, including 2 complex 8; 21 translocations, which have not been reported previously: t (8; 21), der (8) t (8; 21) (8pter --> 8q22::21q22 --> 21qter), der (21) t (8; 21; 8) (8qter --> 8q22:: 21p13 --> 21q22::8q22 --> 8qter) and t (21; 8; 18; 1), der (8) t (8; 21) (8pter --> 8q22:: 21q22 --> 21qter), der (21) t (21; 8; 18; 1) (21p13 --> 21q22?::8q22 --> 8q24 ?:: 18??::1q??q??). The complex karyotypic abnormalities involved nearly all chromosomes, of which the chromosomes 17, 7 and 5 were more involved than the rest. CONCLUSION: Multiplex FISH in combination with conventional cytogenetics may characterize the complex chromosomal abnormalities more precisely. Introduction of this technique to the study of AML with complex chromosomal abnormalities is warranted.

[Expression of Smad4 in leukemia cells].

Loss of transforming growth factor (TGF)-beta signaling has been implicated in malignant transformation of various tissues. Smad4 plays a central role in the signal transduction of TGF-beta. Deletion or mutation of Smad4 has been described in a number of cancers. This study was aimed to investigate a potential role of Smad4 in leukemia including its expression and location in blast cells. The mononuclear cells were separated from bone marrow of leukemia patients. The samples, blast cells of which were more than 90% in mononuclear cells, were selected. The expression and location of Smad4 protein were analyzed by immunohistochemistry methods. The results showed that the Smad4 protein located mainly in nucleus, part of this protein located in cytoplasma, the expressions of Smad4 were not detected in 6 out of 9 ALL patients, in 7 out of 24 AML patients and in 1 out of 2 CML patients; these leukemia patients, in whose cells the expression of Smad4 was not detected, included one L1 and one L3, four L2, one M0, one M1, two M2a, one M3a, one M4b, one M6 and one CML. In conclusion, the Smad4 protein was mainly in nucleus, the deletion or functional change of Smad4 may related with the pathogenesis of human AML.

[Detection of the complex chromosomal aberrations in acute lymphoblastic leukemia by means of multiplex fluorescence in situ hybridization].

This study was aimed to establish the technique of multiplex fluorescence in situ hybridization (M-FISH) and to explore its usefulness in detection of complex chromosomal aberrations (CCAs) in acute lymphoblastic leukemia (ALL). Five ALL patients with CCAs were analyzed by combining the techniques of conventional cytogenetics (CC) and M-FISH. The results demonstrated that M-FISH confirmed the aberrations previously detected by CC, such as t (9;22), t (1;19) and t (y;1), and revealed new abnormalities as der (1) (1::3::7), der (6) t (6;9) (q?;p13), der (1) t (1;11), der (12) t (1;12), der (3) t (3;5), der (2) t (2;16), der (9) (9::18::7) and der (7) (9::18::7), and also corrected the wrong results in CC. Among these abnormalities, der (9) (9::18::7) and der (7) (9::18::7) were reported for the first time. In conclusion, M-FISH has proved to be useful in characterization of the CCAs in ALL, and it is an essential method to refine the karyotype analysis.

[RT-PCR detecting NUP98-HOX fusion gene in leukemia].

To investigate whether there are NUP98-HOXA, NUP98-HOXB, NUP98-HOXC, NUP98-HOXD fusion genes in leukemia patients in Xinjiang, cellular total RNA was extracted from the bone marrow mononuclear cells, the formaldehyde-agarose gel electrophoresis was used to judge whether RNA was intact, the 17 RT-PCR primers were designed to amplify the predicted fusion junctions and 412 bp GAPDH was used as an internal control, NUP98-HOXA fusion genes were amplified by nested-PCR following reverse transcription. One-step PCR was performed to amplify the other predicted fusion genes. The results showed that RNA was proved to be intact and expression of GAPDH was found in every sample. However, no predicted fusion transcripts were detected in leukemia patients. In conclusion, no NUP98-HOX fusion genes were detected in the samples from Xinjiang.