OBJECTIVETo observe the intervening effects of Xuezhikang Capsule (XZK) on levels of blood lipid and other related indices in patients with different Chinese medical syndrome patterns of non-alcoholic fatty liver disease complicated carotid atherosclerosis (NAFLD-CAS), and to seek out the most appropriate pattern to indicate XZK for making guidance of its utilization.

METHODSChinese medical syndrome in 74 patients of NAFLD-CAS were classified into 4 patterns, 34 of Pi-deficiency phlegm-dampness pattern (A), 24 of dampness-heat accumulation pattern (B), 12 of phlegm-stasis intertwined pattern (C), and 4 of Gan-Shen yin-deficiency pattern (D). Excepting those of pattern D were excluded due to too small samples, all patients were treated with XZK for 3 months. Blood levels of blood lipids, including triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C), as well as high-sensitivity C-reactive protein (hs-CRP), and tumor necrosis factor alpha (TNF-alpha) were detected and compared before and after treatment.

RESULTSThe effective rate of XZK on patients of the three patterns, in A-C order, was 97.06%, 91.67%, 91.67%, respectively, with the optimal overall efficacy showed on pattern A. All the indices detected significantly decreased after treatment in all three patterns (P < 0.01), among them, excepting the difference of TG level between groups showed no significance (P > 0.05), the decrements of others were more significant in pattern A than in other two patterns (P < 0.05 or P < 0.01).

CONCLUSIONXZK could reduce the levels of blood lipids, hs-CRP and TNF-alpha in NAFLD-CAS patients, and the Pi-deficiency phlegm-dampness syndrome pattern was the optimal indication of XZK treatment.

Aged ; Carotid Artery Diseases ; complications ; diagnosis ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Liver ; complications ; diagnosis ; drug therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; Phytotherapy ; Treatment Outcome

OBJECTIVEThis study was aimed to detect the expression of HIF-1α in acute myeloid leukemia (AML) except acute promyelocyte leukemia (APL) and investigate the relationship of its expression levels with clinical parameters and prognosis.

METHODSThe primary AML cells were collected from peripheral blood of 53 newly diagnosed AML patients by using CD3 negative sorting. The expression of HIF-1α was measured by real-time fluorescent quantitative PCR (FQ-PCR) , and the relationship between expression level of HIF-1α and clinical parameters (age, sex, WBC count, clinical typing, prognosis) was analysed according to relative expression level. Furthermore, Western blot was used to detect the protein level of HIF-1α in AML patients with or without extramedullary infiltration.

RESULTSThe expression level of HIF-1α did not correlate with age, sex, WBC count, Hb level, Plt count and the percentage of blast. There was no significant difference of HIF-1α expression between different AML subtype based on FAB. The higher level of HIF-1α was found in AML patients who did not get complete remission after one or two courses of chemotherapy, however, the difference was not statistically significant. The relapse rate was higher in AML patients with the higher expression of HIF-1α. In addition, the higher level of HIF-1α mRNA and protein were found in bone marrow of AML patients with extramedullary infiltration (P < 0.01). The negative correlation between HIF-1α and PTEN was observed (r = -0.48, P = 0.001).

CONCLUSIONSOverexpression of HIF-1α are closely related with extramedullary infiltration and prognosis of acute myeloid leukemia, and may be used as an early indicator of extramedullary infiltration and prognosis.

Granulocyte Precursor Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; Leukemia, Myeloid, Acute ; Prognosis ; RNA, Messenger ; Recurrence ; Remission Induction

Objective: To establish a method for determining the contents of quercetin, kaempferide, and isorhamnetin in Anoectochilus roxburghii and to provide scientific method for quality control of A. roxburghii and its related products. Methods: HPLC was used. Using Lanbo-Kromasil-C18 column, with methanol -0.2% phosphoric acid solution (50:50) as mobile phase; The detection wavelength was 360 nm, flow rate was 1.0 mL/min, and column temperature was 30 ℃. Results: Quercetin, kaempferide, and isorhamnetin showed a good linear relationship in the range of 11.0-165.6 (r = 1.0000), 11.4-170.4 (r = 0.9998), and 10.4-156.0 ng (r = 0.9995). The average recoveries of quercetin, kaempferide, and isorhamnetin were 97.69% (RSD = 1.1%, n = 9), 95.09% (RSD = 1.6%, n = 9), and 95.86% (RSD = 1.7%, n = 9). Conclusion: The method is simple, rapid, accurate, reproducible, and can be used as an ideal method for quality control.

OBJECTIVE: To investigate the effect of remifentanil postconditioning on myocardial hypoxia/reoxygenation (H/R) injury in adults, and to further explore the role of aquaporin-4 (AQP-4) in mediating this effect. METHODS: Trabecular muscles from the right atrial appendage of adults were treated with hypoxia for 90 min, followed by reoxygenation for 120 min. Then, remifentanil 0.01, 0.1 and 1.0 nmol·L-1was infused 10 min before the end of hypoxia until 10 min after the start of reoxygenation. The contractile tension of the trabecular muscles was monitored during the experiment. Western blotting was performed to evaluate AQP-4 expression at the end of the experiment. RESULTS: Compared with normal control group, muscle tension decreased significantly after 30 min of induction in H/R group (P<0.05), and it was reduced to the minimum at 90 min. The muscle tension at 150 min and 180 min in remifentanil 0.1 nmol·L-1group was higher than that in H/R group (P<0.05). The muscle tension in remifentanil 1.0 nmol·L-1group was higher than that in H/R group during reoxygenation (P<0.05). Compared with normal control group, the expression of AQP-4 protein in H/R group was significantly higher (P< 0.05), whereas the expression of AQP-4 in remifentanil 0.1 and 1.0 nmol·L-1groups was down-regulated compared with H/R group (P<0.05). CONCLUSION: Remifentanil can alleviate myocardial H/R injury in adults, and its effect may be related to the decreased expression of AQP-4.

BACKGROUNDAmyloid β (Aβ) has been established as a key factor for the pathological changes in the brains of patients with Alzheimer's disease (AD), and cellular senescence is closely associated with aging and cognitive impairment. However, it remains blurred whether, in the AD brains, Aβ accelerates the neuronal senescence and whether this senescence, in turn, impairs the cognitive function. This study aimed to explore the expression of senescence-associated genes in the hippocampal tissue from young to aged 5XFAD mice and their age-matched wild type (WT) mice to determine whether senescent neurons are present in the transgenic AD mouse model.

METHODSThe 5XFAD mice and age-matched wild type mice, both raised from 1 to 18 months, were enrolled in the study. The senescence-associated genes in the hippocampus were analyzed and differentially expressed genes (DEGs) were screened by quantitative real-time polymerase chain reaction. Cognitive performance of the mice was evaluated by Y-maze and Morris water maze tests. Oligomeric Aβ (oAβ) (1-42) was applied to culture primary neurons to simulate the in vivo manifestation. Aging-related proteins were detected by Western blotting analysis and immunofluorescence.

RESULTSIn 5XFAD mice, of all the DEGs, the senescence-associated marker p16 was most significantly increased, even at the early age. It was mainly localized in neurons, with a marginal expression in astrocytes (labeled as glutamine synthetase), nil expression in activated microglia (labeled as Iba1), and negatively correlated with the spatial cognitive impairments of 5XFAD mice. oAβ (1-42) induced the production of senescence-related protein p16, but not p53 in vitro, which was in line with the in vivo manifestation.

CONCLUSIONSoAβ-accelerated neuronal senescence may be associated with the cognitive impairment in 5XFAD mice. Senescence-associated marker p16 can serve as an indicator to estimate the cognitive prognosis for AD population.

Alzheimer Disease ; metabolism ; physiopathology ; Amyloid Precursor Protein Secretases ; genetics ; metabolism ; Amyloid beta-Peptides ; metabolism ; Amyloid beta-Protein Precursor ; metabolism ; Animals ; Aspartic Acid Endopeptidases ; genetics ; metabolism ; Brain ; metabolism ; physiopathology ; Cells, Cultured ; Cellular Senescence ; genetics ; physiology ; Cognition ; physiology ; Cognition Disorders ; metabolism ; physiopathology ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Neurons ; metabolism ; pathology ; Real-Time Polymerase Chain Reaction

This study was aimed to investigate the expression of granulocyte colony-stimulating factor receptor IV(G-CSFR IV) in adult acute leukemia patients and its clinical significance. The bone marrow hematopoietic stem cells from healthy persons were used as controls. The real-time RT-PCR was used to determine the expression level of G-CSFR I-IV in 99 AML, 34 ALL patients and 19 healthy persons. The results showed that the relative expression level of G-CSFR IV/G-CSFR I in AML patients was obviously elevated, as compared with that in ALL patients and controls, while the relative expression level of G-CSFR IV/G-CSFR I in ALL patients showed no statistical difference from controls. The analysis of clinical features and chemotherapeutic efficacy demonstrated that the clinical remission rate in patients with high expression of G-CSFR IV/G-CSFR I was lower than that in patients with low expression. The relative expression level of G-CSFR IV/G-CSFR I was not related with risk stratification from sex, age, blast ratio, FAB typing, chromosome and fusion gene. It is concluded that the abnormal high expression of G-CSFR IV relates with poor prognosis of AML.
Case-Control Studies ; Hematopoietic Stem Cells ; metabolism ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; Protein Isoforms ; metabolism ; Receptors, Granulocyte Colony-Stimulating Factor ; metabolism

OBJECTIVETo establish the mouse model for the expression of PD-L1 by hydrodynamic injection and to study the effects of myeloablative conditioning on hydrodynamic injection-mediated PD-L1 expression.

METHODSPlasmid amplification, hydrodynamic injection, collagenase perfusion, real time PCR, ELISA and flow cytometry were applied to test the expression and function of PD-L1. Also, animal models were set up to test the effects of chemical or radiactive myeloablative conditioning on hydrodynamic injection-mediated PD-L1 expression.

RESULTSThe expression of PD-L1 mRNA and protein could be detected as early as 8 h after hyrodynamic injection and reached peak expression by 24 h, and returned to baseline level by 7 d after injection. Serum PD-L1 level reached to 100 µg/ml as early as 24 h after injection and plateaued at 7 d after injection. Serum PD-L1 persisted for 3 weeks and declined to baseline after 1 month of hydrodynamic injection. The PD-L1 function induced by hydrodynamic injection was consistent with literature reports. At each time point, the PD-L1 expression was not different significantly between the myeloablative conditioning group and control group; the mice transfected with PD-L1 showed a higher survival rate than that in control group.

CONCLUSIONMyeloablative conditioning does not affect hydrodynamic injection-mediated PD-L1 expression, indicating that the PD-L1 can be used in HSCT mouse model.

Animals ; B7-H1 Antigen ; pharmacology ; Disease Models, Animal ; Flow Cytometry ; Hydrodynamics ; Injections ; Mice ; Myeloablative Agonists ; pharmacology ; RNA, Messenger ; Transfection ; Transplantation Conditioning

OBJECTIVETo detect the expression level and the mutantion of CD131 in acute myeloid leukemic (AML) cells, and to analyze the relationship of CD131 expression level with clinical features.

METHODSThe peripheral blood mononuclear cells (PBMNC) from 44 AML patients and 25 healthy donors were collected, and the expression level of CD131 mRNA was detected by RT-PCR. The full length coding sequence of CD131 from 15 patients and 5 healthy donors was amplified by RT-PCR, and linked to pGEM-T vector to clone TA, 10 positive recombinant clones of each sample were analyzed by DNA sequencing. The AML patients were divided into CD131 negative and CD131 positive groups according the expression level of CD131, and white blood cell counts, CD34(+) cells percentage, complete remission rate of patients were compared.

RESULTSCD131 was expressed at a lower level in AML than that in healthy donor. Five kinds of CD131 mutantions could be detected in both AML and healthy donor groups, but the mutation rate in AML (75.33%) was higher than that in healthy donors (18.0%). CD131 negative group showed a higher CD34(+) cells percentage (69.1% ± 20.8%), and lower complete remission rate (33.3%) than that in CD131 positive group (69.1% ± 20.8%, 33.3%). No statistically significant difference of WBC counts was found between 2 groups.

CONCLUSIONCD131 is expressed at a low level and shows high frequency of mutation in acute myeloid leukemic cells. CD131 negative AML correlats with high proportion of CD34(+) cells, and showed insensitive to chemotherapy.

Cytokine Receptor Common beta Subunit ; Humans ; Leukemia, Myeloid, Acute ; Leukocyte Count ; Leukocytes, Mononuclear ; Mutation ; RNA, Messenger ; Remission Induction

OBJECTIVETo investigate the hematologic characteristics and gene diagnosis of patients with Thailand deleted α-thalassemia 1, so as to provide the information for clinical genetic counseling.

METHODSThe clinical data of 32 patients with Thailand delated α-thalassemia 1 were analyzed retrospectively; the hematologic characteristics and gene diagnosis of Thailand deleted type were investigated by using routine hematologic examination, genetic detection of common thalassemia and Thailand deleted α-thalassemia 1.

RESULTSAmong 32 cases, the Thailand deleted α-thalassemia 1 heterozygote was found in 29 cases, the Thailand deleted α-thalassemia 1 and α(3.7) gene deletion double heterozygote were found in 1 case, the Thailand deleted α-thalassemia 1 with β-thalassemia (1 case with codons 41-42 mutation heterozygous, 1 case with CD17 mutation heterozygous) was found in 2 cases by detection. The MCV and MCH levels were decreased in all cases of Thailand deleted thalassemia 1, there were significant differences in RBC, MCV, MCH (P<0.05) between normal control and Thailand deletion α-thalassemia 1 group; there were also significant differences in MCHC (P<0.05) between Southeast asia thalassemia and Thailand deleted α-thalassemia 1 group.

CONCLUSIONThere are no significant differences in hematological parameters except MCHC between Southeast asia thalassemia and Thailand deleted α-thalassemia 1 group. moreover the Thailand deleted α-thalassemia 1 in a certain proportion exists in area with high incidence of thalassemia, therefor the clinicians should pay more attention to the screen and diagnosis of Thailand delated α-thalassemia and can exactly diagnose the Thailand delected α-thalassemia 1 on the basis of comprehensive analysis of conventional and Thailand delected α-thalassemia 1 detection results, clinical presentation, hematologic parameters and ultrasonic examination, so as to avoid the birth of child with severe and intermidiate type α-thalassemia caused by Thailand deleted α-thalassemia 1.

Gene Deletion ; Heterozygote ; Humans ; Mutation ; Phenotype ; Thailand ; alpha-Thalassemia ; beta-Thalassemia

BACKGROUNDAmyloid β (Aβ) deposits and the endoplasmic reticulum stress (ERS) are both well established in the development and progression of Alzheimer's disease (AD). However, the mechanism and role of Aβ-induced ERS in AD-associated pathological progression remain to be elucidated.

METHODSThe five familial AD (5×FAD) mice and wild-type (WT) mice aged 2, 7, and 12 months were used in the present study. Morris water maze test was used to evaluate their cognitive performance. Immunofluorescence and Western blot analyses were used to examine the dynamic changes of pro-apoptotic (CCAAT/enhancer-binding protein homologous protein [CHOP] and cleaved caspase-12) and anti-apoptotic factors (chaperone glucose-regulated protein [GRP] 78 and endoplasmic reticulum-associated protein degradation-associated ubiquitin ligase synovial apoptosis inhibitor 1 [SYVN1]) in the ERS-associated unfolded protein response (UPR) pathway.

RESULTSCompared with age-matched WT mice, 5×FAD mice showed higher cleaved caspase-3, lower neuron-positive staining at the age of 12 months, but earlier cognitive deficit at the age of 7 months (all P < 0.05). Interestingly, for 2-month-old 5×FAD mice, the related proteins involved in the ERS-associated UPR pathway, including CHOP, cleaved caspase-12, GRP 78, and SYVN1, were significantly increased when compared with those in age-matched WT mice (all P < 0.05). Moreover, ERS occurred mainly in neurons, not in astrocytes.

CONCLUSIONSThese findings suggest that compared with those of age-matched WT mice, ERS-associated pro-apoptotic and anti-apoptotic proteins are upregulated in 2-month-old 5×FAD mice, consistent with intracellular Aβ aggregation in neurons.

Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; metabolism ; Animals ; Apoptosis ; physiology ; Blotting, Western ; Caspase 12 ; metabolism ; Endoplasmic Reticulum Stress ; physiology ; Frontal Lobe ; metabolism ; Heat-Shock Proteins ; metabolism ; Immunohistochemistry ; Mice ; Mice, Transgenic ; Neurons ; metabolism ; Transcription Factor CHOP ; metabolism ; Ubiquitin-Protein Ligases ; metabolism ; Unfolded Protein Response ; physiology