The overall action molecular mechanism of anti-hepatitis B active extracts in Flos chrysanthemi indici based on epigenetics and metabonomics / 药学学报

Using the concepts and methods of epigenetics and metabolomics, to investigate the overall action molecular mechanism of Chrysanthemi indici C (CIC), the anti-hepatitis B virus (HBV) active extracts from Flos chrysanthemi indici. The inhibitory effects of CIC on proliferation and hepatitis B surface antigen (HBsAg), hepatitis B envelope antigen (HBeAg) and HBV-DNA of HepG2.2.15 cells were detected by CCK-8 and antigen kit. The DNA methyltransferases (DNMTs)/ten-eleven-translocation-2 (TET2) equilibrium was detected by ELISA. Illumina 850K methylation chip, pyrosequencing and qPCR were used to determine the action pathway and target of CIC by GO and KEGG analysis. Cell metabolites were extracted with 80% methanol, and the changes of differential metabolites, differential metabolic pathways and cell microenvironment were detected by LC-MS and other metabolomics methods. The results showed that CIC could inhibit the proliferation, HBsAg, HBeAg and HBV-DNA of HepG2.2.15 cells obviously, down-regulate DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3a (DNMT3a) and DNA methyltransferase 3b (DNMT3b), up-regulate TET2, and restore the balance of DNMTs/TET2. The action targets of CIC were phospholipase C gamma 2 (PLCG2), phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3), 1-acylglycerol-3-phosphate O-acyltransferase 2 (AGPAT2), 5-hydroxytryptamine receptor 2B (HTR2B), nerve growth factor (NGF), mainly involved in lipid metabolism, inflammation mediated regulation of transient receptor potential (TRP), phospholipase D signaling and advanced glycation end product-receptor for AGE (AGE-RAGE) signaling in diabetic complications pathways. CIC could significantly affect fatty acid metabolism and had great influence on phenolic acid, alkaloid and lipid metabolites in cell microenvironment. These results suggest that the action mechanism of CIC may be the synergistic action of multiple pathways and multiple targets, including related inflammatory pathways, immune pathways and lipid metabolism, through regulating epigenetic expression balance and restoring the balance of cell microenvironment.

Gene Mutation and Overexpression of Newly Diagnosed Multiple Myeloma Patients / 中国实验血液学杂志

OBJECTIVE@#To analyze the characteristics of gene mutation and overexpression in newly diagnosed multiple myeloma (NDMM) patients.@*METHODS@#Bone marrow cells from 208 NDMM patients were collected and analyzed. The gene mutation of 28 genes and overexpression of 6 genes was detected by DNA sequencing. Chromosome structure abnormalities were detected by fluorescence in situ hybridization (FISH).@*RESULTS@#Gene mutations were detected in 61 (29.33%) NDMM patients. Some mutations occurred in 5 or more cases, such as NRAS, PRDM1, FAM46C, MYC, CCND1, LTB, DIS3, KRAS, and CRBN. Overexpression of six genes (CCND1, CCND3, BCL-2, CCND2, FGFR3, and MYC) were detected in 83 (39.9%) patients, and cell cycle regulation gene was the most common. Single nucleotide polymorphisms (SNP) changes were detected in 169 (81.25%) patients, the TP53 P72R gene SNP (70.17%) was the most common. Abnormality in chromosome structure was correlated to gene overexpression. Compared to the patients with normal chromosome structure, patients with 14q32 deletion showed higher proportion of CCND1 overexpression. Similarly, patients with 13q14 deletion showed higher proportion of FGFR3 overexpression, whereas patients with 1q21 amplification showed higher proportion of CCND2, BCL-2 and FGFR3 overexpression.@*CONCLUSION@#There are multiple gene mutations and overexpression in NDMM. However, there is no dominated single mutation or overexpression of genes. The most common gene mutations are those in the RAS/MAPK pathway and the genes of cyclin family CCND are overexpression.

Phacoemulsification in the anterior chamber: alternative surgical technique in post-vitrectomy cataract

Objective: To evaluate the characteristics, safety and effectiveness of a modified technique of phacoemulsification in post-vitrectomy cataracts

Methods: This retrospective clinical trial comprised 31 patients [31 eyes] with post-vitrectomy cataract, who had undergone phacoemulsification combined with intraocular lens implantation. An alternative surgical technique known as phacoemulsification in the anterior chamber was used for nucleus management in those cases. The following parameters were evaluated: best corrected visual acuity [BCVA], ocular inflammation, intraocular pressure, endothelial cell count and surgical complications

Results: Three months after surgery, the BCVA improved significantly compared with that before surgery [Z=-10.547; p<0.05]. There were no significant differences in IOP before and after surgery [Z=-0.474; p>0.05]. There was a statistically significant postoperative decrease in endothelial cell densities [Z=-3.916; p<0.05]. The mean endothelial cell loss was -8.71%. A little inflammatory response in the anterior chamber in 11 eyes and mild corneal edema in 8 eyes were observed on the first day after surgery, which subsided after a week. The posterior capsular opacification were observed in three eyes, two of which were denser, and the YAG laser was performed for posterior capsular incision. No obvious surgical complications occurred

Conclusion: The modified technique of phacoemulsification, with phacoemulsification in the anterior chamber, is safe and effective to deal with post-vitrectomy cataracts

Effects of SUMO specific protease 1 on hPXR-mediated P-gp gene expression / 药学学报

The study aimed to investigate the effects of small ubiquitin-related modifier (SUMO) specific protease 1 (SENP1) on human PXR-mediated MDR1 transcriptional activity and mRNA expression. Empty vector and expression plasmids, including PXR, SENP1 and SENP1 mutant (SENP1m) were transiently transfected into HepG2 and LS174T cells using Lipo2000. Transcriptional activity was detected by dual luciferase reporter gene assay, and mRNA level was measured using real-time polymerase chain reaction. The results showed that SENP1 could remarkably reduce the rifampicin (RIF)-induced MDR1 reporter activity and mRNA level in hPXR over expressed HepG2 and LS174T cells (P < 0.05), whereas adding SENP1m restored the RIF-induced increases (P < 0.05). These results indicated that SENP1 could repress the RIF-induced hPXR-mediated MDR1 transcriptional activity and mRNA expression.

Aplastic anemia with macrocytic anemia: a study based on long-term follow-up / 中华血液学杂志

<p><b>OBJECTIVE</b>To elucidate the clinical features, response rate, prognosis and clonal evolution of aplastic anemia (AA) with macrocytic anemia (mAA).</p><p><b>METHODS</b>The clinical features at initial diagnosis and data in follow up of mAA hospitalized from January 2000 to October 2011 were analyzed retrospectively.</p><p><b>RESULTS</b>(1) Of 153/568 (26.9%) cases of mAA at initial diagnosis, 114(74.5%)were non-severe AA (NSAA), 39(25.5%)severe AA (SAA) and 0 very severe AA (VSAA), while the proportion was 16.2%, 45.2%, and 38.6% in 376 normocytic anemia AA (nAA), and the difference is statistically significant(χ(2) = 181.390; P = 0.000). The median age of mAA was significantly higher than that of nAA \[30(4 - 70)years vs 19 (3 - 68) years, P = 0.001\]. (2) There were no statistical difference in hemoglobin, absolute neutrophil count (ANC), platelet count (PLT), response rate after 6 months treatment and overall survival (OS) between mAA and nAA grouped in SAA and NSAA respectively. In SAA, the reticulocyte count (Ret) of mAA was significantly higher than that of nAA \[23.90(2.99 - 61.00)×10(9)/L vs 13.1(0 - 70.60)×10(9)/L, P = 0.000\] and the proportion of erythroid cells in bone marrow of mAA was also higher \[23.5 (0 - 58) vs 14.5 (0 - 65), P = 0.043\], while they did not differ significantly in NSAA. (3) The proportion of AA with PNH clones or abnormal cytogenetics did not differ significantly in mAA and nAA groups before treatment. The incidences of AA evolved to PNH in mAA and nAA was not statistically significant (7/153 vs 9/376, χ(2) = 1.099, P = 0.294) and so was the incidence of evolution to MDS/AML(3/153 vs 13/376, χ(2) = 0.399, P = 0.528).</p><p><b>CONCLUSION</b>In presented with macrocytic anemia at initial diagnosis of AA, higher proportion of NSAA, elderly age, higher Ret and proportion of erythroid cells are features, but being no statistical difference in the response rate, OS, and proportion of clonal evolution.</p>

The clinical significance of evolution of paroxysmal nocturnal haemoglobinuria clones in aplastic anemia patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical significance of evolution of paroxysmal nocturnal hemoglobinuria (PNH) clones in aplastic anemia (AA) patients.</p><p><b>METHODS</b>The positive rate of PNH clones in 678 AA cases at first diagnosis from January 2002 to December 2009 were analyzed, and to compare the response rate and overall survival (OS) between AA patients with or without PNH clones. All patients were sequentially followed-up to assess the incidence rate and risk factors for AA evolving to overt PNH.</p><p><b>RESULTS</b>(1) Of 119/678 (17.6%) AA patients at initial diagnosis presented with PNH clones,the positive rates of PNH clones among non-severe AA (NSAA), severe AA (SAA) and very severe AA (VSAA) were 16.7% (37/ 222), 17.3% (45/260) and 18.9% (37/196), respectively. There was no statistical difference among the three groups. (Chi2 = 0.369; P = 0.832); (2) 678 newly diagnosed AA cases were divided into 5 subgroups according to PNH clones, severity of disease and treatment regimens. There was no statistical difference among the five subgroups regarding 6m-response rate (RR) and OS. (3) Serial follow-up revealed that persistent PNH negative clones were found in 516 (76.1%) cases, and evolved to PNH positive clones after therapy in 43 (6.3%) cases. Persistent PNH positive clones were found in 72 (10.6%) cases, and disappeared the clones after treatment in 47 (6.9%) cases. There was no statistical difference among the four subgroups in terms of the 6m RR (Chi2 = 2.489,P = 0.426) and OS (P = 0.477); (4) 17 out of 678 AA cases (2.5%) evolved to overt PNH and the estimated incidence of evolution to overt PNH was (3.7 +/- 0.9)% at 10 years. The incidences of AA patients with or without PNH clones at initial diagnosis evolved to overt PNH were 3.4% and 2.3%, respectively. There was no statistical difference between the two groups, (Chi2 = 0.111; P = 0.739); and so was found in OS by Kaplan-Meier analysis (P = 0.868). Cox regression model analysis showed that none of the severity of AA, with or without PNH clone at initial diagnosis, treatment regimen and 6m RR was the risk factor for evolution to overt PNH.</p><p><b>CONCLUSION</b>There is no difference between AA patients presented with or without PNH clones at initial diagnosis regarding the RR and prognosis. The appearance of PNH clones in AA is not identified as a risk factor for developing into overt PNH.</p>

Treatment of severe aplastic anemia with intensified immunosuppressive therapy and two different regimens with recombinant human granulocyte colony-stimulating factor: a retrospective study based on long-term follow-up / 中华血液学杂志

<p><b>OBJECTIVE</b>To compare the efficacy and safety of two different regimens with recombinant human granulocyte colony-stimulating factor (rhG-CSF) combined with intensified immunosuppressive therapy (IIST) in severe aplastic anemia (SAA).</p><p><b>METHODS</b>Retrospectively analyzed 176 SAA treated with IIST and rhG-CSF in our hospital from March 1994 to December 2007. Regimen A (Group A, n = 96), rhG-CSF 300 µg/d was initiated on day 31 after IIST and subcutaneously administered 1-3 days a week for 3 months. Regimen B (Group B, n = 80), rhG-CSF was initiated at 5 µg·kg(-1)·d(-1) before IIST until hematologic recovery.</p><p><b>RESULTS</b>(1) The early response rate of Group B (67.5%) was significantly higher than that of Group A (37.5%) (P < 0.01), the interval from IIST to response in Group B was shorter than that in Group A. Moreover, infection-related deaths during first 4 months after IIST were significantly reduced in Group B (6.3%) when compared with Group A (16.7%) (P = 0.034). The cumulative incidence of survival at 4 years in Groups B [(77.7 ± 4.9)%] was also significantly higher than that in Group A [(57.2 ± 5.1)%] (P = 0.006). (2) With regard to 93 refractory patients with no response 4 months after IIST, rhG-CSF therapy was continued in Group B meanwhile stopped in Group A. There were no differences between two groups in terms of survival and the response rates (P = 0.288, 0.066), but there was an increasing risk of evolving into MDS/AML in Group B (22.3%) when compared with Group A (3.71%) (P = 0.023). (3) By multivariate analysis, the severity of disease (P = 0.010, RR = 1.922) and the early response (P < 0.01, RR = 5.749) were associated with the overall survival. Moreover, the number of days of rhG-CSF therapy was the only significant risk factor for SAA evolving into MDS/AML (P = 0.017, RR = 1.004).</p><p><b>CONCLUSIONS</b>The early initiation of rhG-CSF therapy with proper dose might contribute to a desirable early response and reduced infection-related death rate, but extended administration of rhG-CSF did not improve the long-term outcome of refractory SAA and may further facilitate the progression of SAA into MDS/AML.</p>

Study on clonal evolution of monosomy 7 in patients with aplastic anemia by interphase- fluorescence in situ hybridization / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the clonal evolution of monosomy 7 in patients with aplastic anemia (AA).</p><p><b>METHODS</b>Monosomy 7 (-7) in 81 AA patients with normal karyotype at diagnosis and 46 AA treated with immunosuppressive therapy (IST) and more than 6 months of recombinant human granulocyte colony-stimulating factor (rhuG-CSF) were detected by interphase- fluorescence in situ hybridization (FISH) retrospectively.</p><p><b>RESULTS</b>There were 5.4% - 7.6% of -7 cells in 11 (13.6%) of 81 patients at diagnosis, the survival and response rate to IST in -7 positive patients did not differ significantly from that in -7 negative patients (P = 0.481, 0.865); -7 cells disappeared after IST in all of the 11 patients including 5 received long-term rhuG-CSF therapy, and none of them evolved to myelodysplastic syndromes/acute myeloid leukemia (MDS/AML) at a median follow-up of 44 months. Serial assessments of -7 clones were performed in 46 patients, none of whom detected -7 clones 3-6 months after IST, but -7 recurrence in 5 patients 12 - 15 months after IST. At a median follow-up of 48 months, FISH identified 6 patients with -7 clones while the conventional cytogenetic analysis (CCA) recognized in 5. Moreover, the first demonstration of -7 by FISH was 3 - 18 months earlier than that by CCA. All of the 6 patients with FISH detected -7 evolved to MDS/AML with -7 and four of them were retrospectively analysed for in samples at -7 diagnosis of AA, but none of them was positive.</p><p><b>CONCLUSIONS</b>Monosomy 7 exists in a part of AA patients, but the preexisting -7 cells seems neither associated with fatality nor evolvation to MDS/AML. rhuG-CSF might facilitate the expansion of -7 clones; It is necessary to monitor -7 in AA, especially when received long-term rhuG-CSF therapy.</p>

Diagnostic value of magnetocardiography in patients with coronary heart disease and in-stent restenosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In-stent restenosis (ISR) has become one of the most challenging problems in patients with coronary heart disease. At present, using non-invasive methods to assess ISR is a hot topic. In this investigation we attempted to explore the potential of magnetocardiography (MCG) in diagnosis of in-stent restenosis.</p><p><b>METHODS</b>MCG was analyzed in 52 patients with coronary artery disease for three times: before stenting, one month and 7 months after successful intracoronary stenting.</p><p><b>RESULTS</b>The average classification of total maps (ACTM) and the ratio of abnormal maps (RAM) were lower in 1 month after intracoronary stenting compared with that obtained before stent planting (2.91 vs 2.52, 65.74% vs 42.80%, P < 0.01), while complex ventricular excitation index (CVEI) increased from -42.63 to -20.05 (P < 0.01). In ISR subgroup (n = 16), RAM decreased in 1 month after intracoronary stenting compared to it before stenting (68.99% vs 45.26%, P < 0.05). ACTM increased in 7 months compared to that obtained in 1 month after stenting (3.15 vs 2.51, P < 0.05). According to the ROC curve, ACTM showed its unique diagnostic value in restenosis patients. The sensitivity and specificity of ACTM were 80.0%, 69.40%, respectively. Its positive predictive value and negative predictive value were 54.6% and 88.5%, respectively.</p><p><b>CONCLUSIONS</b>After successful intracoronary stenting, most parameters of MCG were improved. ACTM was of prognostic value in diagnosing ISR.</p>

Magnetocardiography changes in coronary artery disease patients with normal or unspecialized resting electrocardiogram / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the magnetocardiography (MCG) changes in coronary artery disease (CAD) patients with normal or unspecific changes in resting electrocardiogram (ECG).</p><p><b>METHODS</b>MCG mapping was performed by MCG-7 (MaGIC, Magiscan GmbH) installed in an unshielded room. All patients underwent ECG and coronary angiogram examinations and patients with normal or unspecified ECG changes and coronary artery narrowing > or = 70% in at least 1-vessel were defined as CAD group (n = 120). Patients with normal coronary angiogram served as control (n = 82). Four parameters: ACTM (average classification of total maps), RAM (ratio of abnormal maps), CVEI (complex ventricular excitation index) and R-max/T-max ratio, were analyzed in CAD and control groups.</p><p><b>RESULTS</b>RAM (62% vs. 35%) and ACTM (2.62 +/- 0.98 vs. 2.29 +/- 0.90, P < 0.05) were significantly higher in CAD group than in control group. CVEI was found in abnormal zone (-100 - 0) in CAD group while in normal zone (0 - 100) in control group. The ratio of Rmax/Tmax in CAD group was also significantly higher in CAD group than in control group (6.41 +/- 3.29 vs. 4.10 +/- 2.00, P < 0.01). ROC curve analysis indicates that RAM, CVEI and Rmax/Tmax ratio were helpful parameters for CAD diagnosis and the diagnostic sensitivity was 67.1%, 65.9% and 64.3%; the specificity was 65.1%, 68.3% and 68.3% respectively.</p><p><b>CONCLUSION</b>MCG was a useful tool for diagnosing chronic myocardium ischemia in CAD patients with normal or unspecific changes resting ECG.</p>