OBJECTIVETo investigate the presence of leukemia stem cells (LSC) in acute myeloid leukemia (AML) and find out the relative position of leukemia cells in the figures of flow cytometry, and to analyze the relationship between minimal residual diseases (MRD) and the level of LSC, so as to explore the correlation of LSC changes with the curative effect and the prognosis during chemical therapy.

METHODSA total of 85 samples were collected from 50 AML (except M3) patients, including 50 samples from the newly diagnosed patients, 7 samples of AML patients with non-remission and 28 samples of AML patients with complete remission. All samples were used for detection of LSC from immune phenotype of CD34/CD38/CD123 by flow cytometry. The detection of immune phenotypic of leukemia cells was performed in the newly diagnosed patients. The detection of leukemia- associated immune phenotypes (LAIP) was implemented in the non-newly diagnosed patients.

RESULTSThe LSC was identified in the CD34/ CD38/ CD123 in AML and consistent with the relative position of the leukemia cell in flow cytometry figures. Statistical analysis showed significant difference in LSC content between the newly diagnosed AML group and the post-chemotherapy complete remission group(P<0.01),but did not between the newly diagnosed AML group and the post-chemotherapy non-remission group(P>0.05).There was significant positive correlation between the LSC content and MRD level in 28 AML patients with complete remission (r=0.680,P<0.01).

CONCLUSIONLSC exist in AML and the relative position are consistent with the leukemia cells in flow cytometry figures, the size characteristics and weak expression of CD45 are also similar to leukemia cells. The proportion of LSC decreases after chemotherapy. Detecting and tracking the LSC changes in bone marrow and combination with detecting minimal resident disease(MRD) may contribute to evaluate the theraputic efficacy and prognosis of leukemia patients.

Flow Cytometry ; Humans ; Interleukin-3 Receptor alpha Subunit ; Leukemia, Myeloid, Acute ; Neoplasm, Residual ; Neoplastic Stem Cells ; Prognosis

Objective:To study diagnostic value of combined detection of heart type fatty acid binding protein (H-FABP) and brain natriuretic peptide (BNP) for chronic heart failure (CHF) in aged patients.Methods:A total of 178 aged CHF patients hospitalized in our hospital from Jul 2015 to Jun 2016 were selected.According to NYHA cardiac function classification,they were divided into class Ⅱ group (n＝70) and class Ⅲ ～ Ⅳ group (n＝48).An-other 52 aged healthy subjects undergoing physical examination were selected as healthy control group simultaneous-ly.Plasma BNP,H-FABP levels,LAD,LVESd,LVEDd,IVST and LVEF were measured and compared among three groups.Correlation among plasma BNP,H-FABP levels and above cardiac function indexes were analyzed.Results:Compared with healthy control group,there were significant rise in plasma levels of BNP [ (50.65 ± 5.86) ng/L vs.(212.65 ± 17.12) ng/L vs.(1264.73 ± 119.74) ng/L] and H-FABP [ (1.68 ± 0.36) ng/ml vs.(3.34 ± 0.56) ng/ml vs.(9.64 ± 1.27) ng/ml],LAD [ (3.25 ± 0.29) cm vs.(4.98 ± 0.23) cm vs.(5.48 ± 0.51) cm],LVESd [ (28.68 ± 2.7) mm vs.(40.98 ± 4.55) mm vs.(58.63 ± 5.75) mm],LVEDd [ (43.22 ± 4.29) mm vs.(56.63 ± 5.23) mm vs.(61.33 ± 6.07) mm] and IVST [ (8.15 ± 0.68) mm vs.(11.87 ± 1.03) mm vs.(15.93 ± 1.43) mm],and significant reduction in LVEF [ (64.84 ± 6.39)% vs.(49.76 ± 4.76)% vs.(35.76 ± 3.62)%] in NYHA class Ⅱ group and NYHA class Ⅲ ～ Ⅳ group;compared with NYHA class Ⅱ group,there were significant rise in plasma levels of BNP and H-FABP,LAD,LVESd,LVEDd and IVST,and significant reduction in LVEF in NYHA class Ⅲ ～ Ⅳ group,P＝0.001 all.Spearman correlation analysis indicated that plasma BNP,H-FABP levels were significant positively correlated with LAD,LVESd,LVEDd and IVST (r＝0.53～0.71,P＝0.001 all),and significant inversely correlated with LVEF (r＝ -0.54,-0.65,P＝0.001 both);plasma BNP level was signifi-cantly positive correlated with H-FABP level (r＝0.36,P＝0.04).Conclusion:Combined detection of BNP and H-FABP contributes to diagnosing CHF in aged patients and assessing its severity.

Objective:To evaluate the efficacy and safety of dl-3-butylphthalide ( NBP) injection and soft capsules in the treat-ment of acute middle cerebral artery infarction. Methods:Sixty-one patients with acute cerebral infarction in the left middle cerebral artery in 72 hours of onset of ischemic stroke with score of 5-25 according to the national institutes of health stroke scale ( NIHSS) were randomly divided into the observation group (n=31) and the control group (n=30). The control group was treated with the routine treatment, while the observation group was sequentially treated with NBP injection and soft capsules additionally. The treatment course was 90 days. Before the treatment, the NIHSS score was evaluated in both groups to compare the neurologic impairment degree. After the treatment, the daily living skills assessment was performed by Barthel index ( BI) and modified Rankin score ( mRS) , and the ad-verse reactions were recorded. Results:Before the treatment, the NIHSS score in the two groups had no statistical significance ( P>0. 05). After the treatment, the BI in the observation group and the control group was (88. 55 ± 16. 74) and (70. 67 ± 26. 18), and mRS was (1. 87 ± 1. 02) and (2. 53 ± 1. 40), respectively, suggesting the observation group had more favorable outcome than the con-trol group (P≤0. 05). The incidence of adverse reactions had no significant difference between the groups. Conclusion: dl-3-Bu-tylphthalide sequential therapy should be regarded as an effective and safe method for acute cerebral infarction, which can improve the daily living skills and 90-day outcome of patients.

To observe the effect of geniposide on non-alcoholic fatty liver disease (NAFLD), and discuss the mechanism of geniposide for NAFLD from the aspect of free fatty acid, forty healthy Wistar male rats were randomly divided into normal group, model group, geniposide and Xuezhikang group. The rats in normal group were fed with normal diets, and the rats in other 3 groups were given with high-fat diet for 8 weeks to induce the NAFLD models. From the week 5 to end of week 8, the rats in geniposide and Xuezhikang group were intervened with corresponding medicines. The body weight, liver wet weight, and fat weight of the rats were recorded. Visual and pathological changes in hepatic tissues were observed with HE staining. The contents of TG, FFA, FAS, AMPK, ACCase and Malonyl-CoA in hepatic tissue, contents of CHO and LDL-C in serum and activities of AST and ALT in serum were detected by using corresponding methods. The results showed that the body weight, liver wet weight, and fat weight of the rats, CHO, LDL-C, ALT and AST levels in serum, TG, FFA, FAS, ACCase and Malonyl-CoA levels in hepatic tissues of the rats in model group were significantly higher than those in normal group (P<0.01), while AMPK activity was significantly lower than that of the normal group (P<0.01), with obvious visual and pathological steatosis in hepatic tissues, and inflammatory injury occurred in model group. Compared with the model group, body weight of the rat, fat weight, levels of FFA in hepatic tissues, ALT and AST activities in serum, liver wet weight, TG, FAS, ACCase and Malonyl-CoA levels were significantly decreased in geniposide group (P<0.01), while the AMPK activity in hepatic tissues was significantly increased (P<0.05),with improvement in visual and pathological performance. Compared with the model group, liver wet weight, fat weight, TG and FFA levels in hepatic tissues, and LDL-C level in serum were significantly decreased in Xuezhikang group (P<0.05). Compared with Xuezhikang group, the body weight of rat, fat weight and FFA level in hepatic tissues were significantly lower in geniposide group (P<0.01), but with no significant difference in other aspects. These findings indicated that geniposide was highly effective in improving the pharmacological effect of NAFLD induced by high-fat diet, and the mechanism was achieved through AMPK-ACCase-Malonyl-CoA-FFA axis.

OBJECTIVETo observe the efficacy of Zaozhu Yinchen Recipe (ZZYCR) on non-alcoholic steatohepatitis (NASH) patients, and to explore its effect on serum free fatty acid (FFA) and tumor necrosis factor alpha (TNF-alpha).

METHODSTotally 120 patients with NASH were randomly assigned to the treatment group (60 cases, treated with ZZYCR, one dose per day) and the control group (60 cases, treated with Silibin Meglumine Tablets, 20 mg each time, thrice per day). The therapeutic course for all was 24 weeks. Serum levels of ALT and AST activities, TC and TG levels were detected before and after treatment. Peritoneal CT was performed in all patients, and CT ratios of liver and spleen calculated. NAFLD activity score (NAS) and degree of hepatic fibrosis were assessed using pathological examinations of liver tissue, and efficacy also evaluated. Serum contents of FFA and TNF-alpha were also detected.

RESULTSCompared with before treatment in the same group, activities of ALT and AST, serum levels of TC, TG, FFA, and TNF-alpha, NAS, scores of symptoms and signs all obviously decreased, degree of hepatic fibrosis was obviously improved in the two groups (P < 0.05, P < 0.01). These changes were more obviously seen in the treatment group (P < 0.05). After 24-week treatment, the total effective rate and total clinical efficacy were 80.00% (48/60 cases) and 85.00% (51/60 cases) in the treatment group, obviously higher than those in the control group [60.00% (36/60 cases) and 73.33% (44/60 cases) respectively], with significant difference (P < 0.05, P < 0.01).

CONCLUSIONZZYCR could improve the clinical efficacy of NASH patients, and its mechanism might be associated with inhibiting serum levels of FFA and TNF-alpha.

Drugs, Chinese Herbal ; therapeutic use ; Fatty Acids, Nonesterified ; blood ; Humans ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

To investigate the effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor simvastatin (SV) on proliferation, apoptosis and the PI3K/AKT signaling pathway in human acute monocytic leukemia cell line SHI-1. SHI-1 cells were incubated with different concentrations of SV (5, 10, 15 µmol/L). Otherwise, SHI-1 cells without any treatment were used as control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detection. MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the early stage apoptosis ratio. The human PI3K-AKT Signaling Pathway RT(2) Profiler(TM) PCR Array was used to detect the expression of 84 genes involved in PI3K-AKT signaling. The results indicated that the SV inhibited the proliferation and inducted the apoptosis of SHI-1 cells in time- and dose-dependent manners significantly. The growth inhibition rates of SHI-1 cells treated with 15 µmol/L SV for 24, 48 and 72 h were 26.82, 47.09 and 63.92, respectively; and their early stage apoptosis ratios were 5.75, 13.25 and 15.59, respectively. Compared with the control group, expression levels of 39 genes were changed in the group of 15 µmol/L SV at 48 h, among them 26 genes were down-regulated and 13 genes were up-regulated. It is concluded that the SV can inhibit proliferation and induce apoptosis of SHI-1 cells, and the mechanism may be associated with the changes of gene expression level in PI3K-AKT signaling pathway regulated by SV.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Monocytic, Acute ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; Simvastatin ; pharmacology

Breast cancer is one of the leading causes of cancer death worldwide. This study aimed to analyze the expression of centromere protein H (CENP-H) in breast cancer and to correlate it with clinicopathologic data, including patient survival. Using reverse transcription-polymerase chain reaction and Western blotting to detect the expression of CENP-H in normal mammary epithelial cells, immortalized mammary epithelial cell lines, and breast cancer cell lines, we observed that the mRNA and protein levels of CENP-H were higher in breast cancer cell lines and in immortalized mammary epithelial cells than in normal mammary epithelial cells. We next examined CENP-H expression in 307 paraffin-embedded archived samples of clinicopathologically characterized breast cancer using immunohistochemistry, and detected high CENP-H expression in 134 (43.6%) samples. Statistical analysis showed that CENP-H expression was related with clinical stage (P = 0.001), T classification (P = 0.032), N classification (P = 0.018), and Ki-67 (P < 0.001). Patients with high CENP-H expression had short overall survival. Multivariate analysis showed that CENP-H expression was an independent prognostic indicator for patient survival. Our results suggest that CENP-H protein is a valuable marker of breast cancer progression and prognosis.
Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Breast ; cytology ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Female ; Follow-Up Studies ; Humans ; Ki-67 Antigen ; metabolism ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Survival Rate ; Up-Regulation

BACKGROUNDEstrogen deficiency contributes to postmenopausal osteoporosis. Periosteum might be a potential target of estrogen, but the underlying mechanism at gene level is far from being elucidated. The objective of this study was to investigate the correlation between estrogen and fatty acid synthase (FAS) expression in periosteum.

METHODSHuman periosteum cells were cultured in vitro. Expressed genes in the substrated cDNA library were verified using semi-quantitative PCR and real-time PCR. The expression of FAS in periosteum of ovarectomized (OVX) SD rats was investigated.

RESULTSFAS gene was most significantly expressed in the subtracted cDNA library of periosteal cells screened by semi-quantitative PCR. Low FAS expression was verified by real-time PCR in the estrogen exposed human periosteum rather than in the control. The estradiol levels were (20.81 +/- 12.62) pg/ml, (19.64 +/- 4.35) pg/ml and (13.47 +/- 1.84) pg/ml in the sham group, the control, and the OVX group, respectively. The estradiol levels in the OVX group was significantly lower (P = 0.0386). The FAS gene expression in periosteum in the OVX group, sham group, and control group was 3.09 +/- 1.97, 1.33 +/- 0.47 and 1.51 +/- 1.32, respectively. The gene expression in the OVX group was significantly higher (P = 0.0372).

CONCLUSIONEstrogen modulates FAS gene expression in in vitro human perisoteum as well as in in vivo rat periosteum.

Animals ; Cells, Cultured ; Estradiol ; blood ; pharmacology ; physiology ; Fatty Acid Synthases ; genetics ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Ovariectomy ; Periosteum ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction