From an aqueous extract of Lonicera japonica flower buds, sixteen compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, MCI gel, silica gel, and sephadex LH-20 and reversed-phase HPLC. Their structures were elucidated by spectroscopic data analysis as 6'-O-acetylvogeloside (1), 6'-O-acetylsecoxyloganin (2), dichlorogelignate (3), guanosinyl-(3' --> 5')-adenosine monophosphate(GpA,4) , 5'-O-methyladenosine (5), 2'-O-methyladenosine (6), adenosine (7), syringin (8), methyl 4-O-β-D-glucopyranosyl caffeate (9), (-)-dihydrophaseic acid 4'-O-β-D-glucopyranoside (10), ketologanin (11), 7α-morroniside (12), 7β-morroniside (13), kingiside (14), cryptochlorogenic acid methyl ester (15), and 6-hydroxymethyl-3-pyridinol (16). All the compounds were obtained from this plant for the first time, compounds 1 and 2 are new compounds, 3 and 5 are new natural products, and 4 is the first example of dinucleoside monophosphate isolated from a plant extract.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flowers ; chemistry ; Lonicera ; chemistry ; Mass Spectrometry ; Molecular Structure

OBJECTIVESTo investigate the effects of blood pressure lowering treatment on stroke recurrence among Chinese patients with previous cerebrovascular diseases.

METHODSPatients were eligible if they had a history of stroke or transient ischaemic attack (TIA) within the previous 5 years. Participants had no definite indication or contraindication for study drugs. There were no blood pressure entry criteria. This study was a large-scale randomised, double-blind, placebo controlled clinical trial. After 4 weeks run-in period, 1520 randomised patients received either ACE inhibitor-perindopril (+ diuretic-indapamide) treatment or matching placebo for 4 years. The primary study outcome was stroke event. Secondary outcomes included cardiovascular death, myocardial infarction, all-cause death and blood pressure.

RESULTSSsven hundred and sixty-two were assigned active treatment and 758 assigned placebo. The characteristics of randomised patients of active and control groups were similar. In active and control groups: 70.8% and 70.5% were male, 93.8% and 93.4% had a history of stroke (haemorrhagic or cerebral infarction). Baseline mean blood pressure in the active and control groups were 145.3 +/- 20.2/86.8 +/- 11.1 mm Hg and 145.3 +/- 20.3/87.2 +/- 10.8 mm Hg, and mean age were 63.9 +/- 7.5 and 63.8 +/- 7.7 years, respectively. Blood pressure in those assigned active treatment was reduced on average 14/6 mm Hg more than placebo at 4 years. During double-blind treatment, active treatment reduced stroke recurrence (8.8% vs 19.4%) by 55% (P < 0.001), myocardial infarction (1.4% vs 2.8%) by 48% (P = 0.070), cardiovascular mortality (3.6% vs 6.6.%) by 45% (P = 0.010), and all-cause mortality (6.3% vs 9.8%) by 36% (P = 0.010). These benefits were achieved similarly in all subgroups: male or female, middle-aged or elderly, with or without hypertensive, cerebral infarct or haemorrhagic stroke.

CONCLUSION4 years of blood pressure lowering treatment was beneficial in Chinese patients with previous cerebrovascular diseases.

Aged ; Antihypertensive Agents ; therapeutic use ; Cerebrovascular Disorders ; drug therapy ; China ; Double-Blind Method ; Female ; Humans ; Male ; Middle Aged ; Randomized Controlled Trials as Topic ; Recurrence ; Stroke ; drug therapy ; prevention & control ; Treatment Outcome

The study was aimed to isolate and establish mesenchymal stem cell line from adult murine bone marrow as well as to identify its biological characteristics and differentiation potential. Bone marrow cells (BMCs) were collected by flushing the femurs and tibias of 4 - 5-week-old male C57BL/6 mice, and were inoculated at a concentration of 1 x 10(6)/cm(2). mMSCs were isolated, enriched and expanded by using bone marrow adherant culture and monoclonal culture. The characteristics of the cells, such as morphology, growth pattern, cell cycle, phenotype, karyotype and multipotent differentiation potential, cytogenetic stability and tumorigenesis were determined. The results indicated that the cell population consisted of spindle- and star-shaped cells, they were highly positive for CD29, CD44, Sca-1, MHC-I, moderate positive for CD13, CD90.2 and negative for CD117, CD45, Flk-1 and MHC-II. mMSCs could be induced to differentiate into adipocytes, osteoblast cells and chondrocytes. It is concluded that mMSC can be isolted, expanded and enriched by using bone marrow adhcrent culture and monoclonal culture. No tumor formations are observed for 3 months in nude mice after subcutaneous injection. mMSCs retain their properties after at least 30 passages in culture as well as from frozen stocks.
Animals ; Cell Differentiation ; physiology ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Hyaluronan Receptors ; metabolism ; Integrin beta1 ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; immunology ; physiology ; Mice ; Mice, Inbred C57BL ; Mice, Nude

OBJECTIVETo explore the clinical and laboratory characteristics of myleodysplastic syndrome (MDS)/myeloproliferative neoplasm (MPN) with PDGFRβ abnormalities.

METHODSChromosome specimens were prepared directly and/or short-time culture of bone marrow cells. Karyotyping was performed with R-binding technique. Fluorescence in situ hybridization (FISH) was performed using PDGFRβ, PDGFRα, FGFR1 break-apart probes and whole chromosome 5 and 12 painting probes, respectively. The expression of JAK2 V617F was measured with quantitative PCR.

RESULTSThe clinical and hematological findings of 27 patients were compatible with diagnosis of MDS/MPN. PDGFRβ rearrangement was detected in 4 patients with D-FISH, and 2 of which were confirmed as t(5;12) by chromosome painting. PDGFRα, FGFR1 and JAK2 V617F mutation were not detected in these 4 PDGFRβ positive MDS/MPN patients with.

CONCLUSIONSPDGFRβ gene rearrangement may be detected in some MDS/MPN patients. FISH is a convenient and reliable approach to detect PDGFRβ gene.

Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Myeloproliferative Disorders ; genetics ; Neoplasms ; Receptor, Platelet-Derived Growth Factor beta ; genetics

OBJECTIVETo analyze the clinical and laboratory characteristics of hematological diseases associated with eosinophilia.

METHODSKaryotype analysis was performed by direct method and/or short-time culture of bone marrow cells for R-banding. Fluorescence in situ hybridization (FISH) was performed using PDGFRα, PDGFRβ and FGFR1 break-apart probes.

RESULTSThe clinical and hematological findings of 44 patients were diagnosed as hematological diseases associated with eosinophilia. Abnormal karyotypes were detected in 6 cases (13.64%) with karyotyping. The efficiency of the detection of abnormal clone was markedly increased to 29.55% (13/44) with FISH techniques, including 7 cases with FIP1L1-PDGFRα (F/P, 15.91%), 3(6.82%) PDGFRα rearrangement, 2 (4.55%) aberrant PDGFRβ gene and 1(2.27%) FGFR1 rearrangement. Patients being PDGFRα, PDGFRβ or FGFR1 positive (13 cases) or negative (31 cases) showed predominant difference in clinical and laboratory features. The incidence of gut involvement, the absolute count of eosinophils in peripheral blood and the percentage of immature eosinophils in bone marrow were significantly increased in positive patients (P < 0.05).

CONCLUSIONSThe hematological diseases associated with eosinophilia are characterized by unique clinical and laboratory features. Karyotyping should be a routine approach to detect the abnormal clone in these diseases. Screening for PDGFRα, PDGFRβ and FGFR1 gene with FISH can provide more genetic information.

Abnormal Karyotype ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; Cytogenetics ; Eosinophilia ; etiology ; genetics ; Female ; Hematologic Diseases ; complications ; genetics ; Humans ; Karyotyping ; Male ; Middle Aged ; Receptor, Platelet-Derived Growth Factor alpha ; genetics ; Young Adult

OBJECTIVETo report a hybrid acute leukemia (HAL) patient with t (12; 22) (p13; q12).

METHODSChromosome specimens were prepared by direct method and/or short-time culture of bone marrow cells. Karyotyping was performed by R-banding technique. Leukemia surface markers were detected by anti-biotin-biotin complex and monoclonal antibodies. Chromosome painting (fluorescence in situ hybridization, FISH) was performed by using whole chromosome 12 and 22 probes labeled with green and red fluorescence, respectively.

RESULTSThe clinical and hematological findings were compatible with the diagnosis of HAL. Lymphoid and myeloid markers were positive on the leukemia cells. Karyotype analysis showed that the patient had t (12; 22) (p13; q12) translocation. A reciprocal translocation between chromosomes 12p and 22q was proved by FISH.

CONCLUSIONSt (12; 22) translocation is a rare chromosome abnormality in leukemia. Patients with t (12; 22) had unique clinical, cytogenetic features. This translocation as a cytogenetic marker for poor-prognosis in leukemia needs to be further studied.

Adult ; Chromosome Banding ; Chromosomes, Human, Pair 12 ; genetics ; Chromosomes, Human, Pair 22 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Leukemia, Biphenotypic, Acute ; diagnosis ; genetics ; Translocation, Genetic

OBJECTIVETo explore the morphologic, immunophenotypic, cytogenetic and clinical features of acute lymphoblastic leukemia (ALL) patients with dicentric (9; 20) (p11 - 13; q11).

METHODSChromosome specimens of bone marrow cells were prepared by direct method and/or short-time culture. Karyo-typing was performed by R-banding technique. Dual-color fluorescence in situ hybridization (FISH) was performed using both chromosome 9 classical satellite probe and chromosome 20 alpha-satellite probe in one patient.

RESULTSThe two ALL patients were positive for CD10 and HLA-DR, showing of B cell origin. Both patients had dicentric (9; 20): case 1 was 45, XY, der (9) t (9; 20) (p11; q11), -20[20]; case 2 was 45, XX, der (9) t (9; 20) (p13; q11), t (9; 22) (q34; q11), -20[10]/46, idem, +8[16]/47, idem, +8, +21[14]. Mutual translocation between chromosomes 9 and 20 of the dicentric chromosome was confirmed by FISH in one patient.

CONCLUSIONSDicentric (9; 20) (p11 - 13; q11) is a rare recurring chromosome abnormality associated with ALL. Because of the subtle nature of the translocation, FISH is essential for the detection of this abnormality.

Adult ; Base Sequence ; Chromosome Banding ; Chromosomes, Human, Pair 20 ; genetics ; Chromosomes, Human, Pair 9 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Molecular Sequence Data ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; pathology ; Sequence Analysis, DNA ; Translocation, Genetic

OBJECTIVETo explore the clinical and laboratory characteristics of two myelodysplastic syndromes (MDS) patients with double isochromosome 20q- anomaly.

METHODSBone marrow cell chromosome preparations were made with both direct method and short-term culture. Karyotype analysis was performed by R-banding technique, and dual-color FISH (fluorescence in situ hybridization) by using a 20q telomeric probe and a sequence-specific probe for 20q12.

RESULTSThe clinical and hematological findings were comparable with diagnosis of MDS. Karyotype analysis showed that both patients had double isochromosome 20q- anomaly: case 1 is 46, XX, der(20)? i(20q-) [6]/46, idem, der (6) i (6p) [1]/47, idem, +der (20)? i (20q-) [3]/47, idem, der(6)i (6p), +der(20)? i (20q-) [20]; case 2 is 45, XY, -7, der (20)? i (20q-) [17]/46, idem, +der(20) ? i(20q-) [3]. Two derivative chromosomes 20 were proved 20q isochromosomes with interstitial deletions by dual-color FISH in one patient.

CONCLUSIONSDouble isochromosome 20q- anomaly is a rare recurrent karyotype abnormality in MDS, and signals a poor prognosis.

Chromosome Banding ; Chromosomes, Human, Pair 20 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Isochromosomes ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics

OBJECTIVETo develop a novel single nucleotide polymorphism (SNP)-PCR based method for quantitative detection of chimerism after allogeneic haemopoietic stem cell transplantation (allo-HSCT), and to explore its feasibility, accuracy and superiority.

METHODS18 SNP loci were sereened to identify informative markers for detecting chimerism in each donor/recipient pair before transplantation. Then the chimerism rate of each informative marker was analyzed by real-time quantitative PCR (RQ-PCR). The accuracy and sensitivity were verified by multiple proportion dilution and analogy chimerism compared with quantitative detection of short tandem repeat (STR)-PCR, fluorescence in situ hybridization (FISH) and fusion gene.

RESULTS(1) The average slope of the 17 time amplications of the internal control plasmid was -3.39, the average intercept was 39.97, correlation coefficients were more than 0.995, which was close to the theoretical level. The intra- and interassay variability was 0.50% and 1.1%, respectively, which were both in the allowed ranges. A linear correlation with artificial mixed chimerism is above 0.99 and a sensitivity of 0.01% proved reproducible. (2) At least one informative marker could be found in over 95% of 40 donor/recipient pairs. The results of the chimerisms derived from SNP-PCR were consistent with that from STR-PCR (96.7%), FISH and fusion gene analasis (P > 0.05); the quantitative results of special fusion gene transcripts were negtive in complete chimerism samples, and positive in mixed chimerism samples.

CONCLUSIONSThis new assay which overcome the PCR competition and plateau biases of STR-PCR provides an accurate, reliable and rapid quantitative assessment of mixed chimerism after allo-transplantation. It is highly promising for of clinical application and may take the place of STR-PCR in the conventional chimerisim assessment.

Chimerism ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; Polymorphism, Single Nucleotide ; Stem Cell Transplantation ; Transplantation Chimera ; Transplantation, Homologous

Objective To analyze the safety of renal transplant from donors with primary central nervous system (CNS) tumors.Methods We retrospectively analyzed the clinical data of 33 donors with primary CNS tumors and the 63 corresponding renal recipients between January 2013 and December 2016 in Tongji Hospital.Results The mean period from diagnosis as primary CNS tumor to donation was about (21.8± 46.4) months (range:0.5 to 192.0 months).The pathological classification of these tumors included gliomas,meningioma,medulloblastoma,etc.Besides,there were 10 donors with high-grade CNS malignancies.Eleven donors have ever been through at least one of the four treatments (craniotomy,V-P/V-A shunt,radiotherapy and chemotherapy),14 donors have undergone none,and the clinical data of rest were unavailable.All the 63 recipients got well renal function after transplant.During an average follow-up of (15.9 ± 8.2) months (range:2.7 to 35.5 months),one recipient got donor-derived rhabdoid tumor 4 months posttransplant,underwent comprehensive treatments,including allograft nephrectomy,radiotherapy,chemotherpy and returned to hemodialysis,while the 62 cases got no donor-derived tumors.Conclusion Tumor transmission of renal allograft from donors with primary CNS tumors is inevitable but with low risk,which means this kind of donors can be used with careful assessment,full informed consent and good balance between wait-list death and tumor transmission.