Study on efficacy and regulation of inflammatory factors with oseltamivir phosphate by different time series of interventions in a murine model of influenza A H1N1 virus infection / 中国药理学通报

Aim To compare the effects of different time sequence interventions on virus infected mice by using oseltamivir (Tamiflu) as a "tool drug" in view of the current situation of the too early the administration time of antiviral in vivo experiment, so as to provide a basis for selecting a reasonable model intervention time point for antiviral drug research. Methods Balb/c mice were randomly divided into six groups. The virus infection model was established by intranasal infection with influenza virus (0.25 TCID50). Tamiflu-1 group and Tamiflu-2 group were administered orally on 1st and 4th day after exposure. The body mass, survival rate, organ index, viral load and inflammatory factor content were measured. Results Compared with the blank control group, the body weight of the mice in the model group decreased and the lung index increased significantly (P < 0.05). The expression levels of 13 inflammatory factors in model 2 group were significantly different ( P < 0.05). Compared with the model-1 group ,the lung index and spleen index of the Tamiflu-1 group decreased significantly (P < 0.05). Compared with the mode-2 group,the lung index in the Tamiflu-2 group was significantly lower (P <0.05) ,and the thy-mus index was significantly higher (P<0.05). The viral load was 0. 03 times that of the model-2 group. The expression levels of 13 inflammatory factors were significantly different (P < 0. 05). Conclusions The symptoms of the mice in Scheme 2 are more obvious and stable after exposure. After administration, the lung inflammation damage is alleviated. Considering the latency, drug intervention is in line with the drug indications when the model animals show symptoms. It will be more reasonable and accurate whether in the model evaluation or drug evaluation.

CCL5-Secreting Virtual Memory CD8+ T Cells Inversely Associate With Viral Reservoir Size in HIV-1-Infected Individuals on Antiretroviral Therapy.

Recent studies highlighted that CD8+ T cells are necessary for restraining reservoir in HIV-1-infected individuals who undergo antiretroviral therapy (ART), whereas the underlying cellular and molecular mechanisms remain largely unknown. Here, we enrolled 60 virologically suppressed HIV-1-infected individuals, to assess the correlations of the effector molecules and phenotypic subsets of CD8+ T cells with HIV-1 DNA and cell-associated unspliced RNA (CA usRNA). We found that the levels of HIV-1 DNA and usRNA correlated positively with the percentage of CCL4+CCL5- CD8+ central memory cells (TCM) while negatively with CCL4-CCL5+ CD8+ terminally differentiated effector memory cells (TEMRA). Moreover, a virtual memory CD8+ T cell (TVM) subset was enriched in CCL4-CCL5+ TEMRA cells and phenotypically distinctive from CCL4+ TCM subset, supported by single-cell RNA-Seq data. Specifically, TVM cells showed superior cytotoxicity potentially driven by T-bet and RUNX3, while CCL4+ TCM subset displayed a suppressive phenotype dominated by JUNB and CREM. In viral inhibition assays, TVM cells inhibited HIV-1 reactivation more effectively than non-TVM CD8+ T cells, which was dependent on CCL5 secretion. Our study highlights CCL5-secreting TVM cells subset as a potential determinant of HIV-1 reservoir size. This might be helpful to design CD8+ T cell-based therapeutic strategies for cure of the disease.

Synchronous conjugation of i-motif DNA and therapeutic siRNA on the vertexes of tetrahedral DNA nanocages for efficient gene silence

The functionality of DNA biomacromolecules has been widely excavated, as therapeutic drugs, carriers, and functionalized modification derivatives. In this study, we developed a series of DNA tetrahedron nanocages (Td),

Synthesis and antitumor activity of α,ß-unsaturated carbonyl moiety- containing oleanolic acid derivatives targeting PI3K/AKT/mTOR signaling pathway.

Oleanolic acid (OA) and its semi-synthetic derivatives have been reported to have a wide range of biological activities. The introduction of electrophilic Michael acceptor group can increase the reactivity of OA to cellular targets and thus improve the anti-tumor activity. In this work, a series of novel α,ß-unsaturated carbonyl derivatives of OA were designed and synthesized. Their in vitro cytotoxic activity against MCF-7, HepG2 and HeLa cells were tested. Most derivatives exhibited improved cell growth inhibitory activity, especially for 3d with an IC50 of 0.77 µM in MCF-7 cells. Moreover, 3d inhibited the migration of MCF-7 and HeLa cells at the concentration of 4 µM. Flow cytometric analysis revealed that 3d induced cell apoptosis and S phase arrest in a concentration-dependent manner. Western blotting experiment demonstrated that 3d inhibited the phosphorylation of AKT and mTOR. These results suggest that this series of OA derivatives bearing exocyclic methylene ketone pharmacophore are promising anticancer agents as potential PI3K/AKT/mTOR pathway inhibitors.

Combination analysis of the physiology and transcriptome provides insights into the mechanism of silver nanoparticles phytotoxicity.

Silver nanoparticles (AgNPs) have adverse impacts on plants when released into environments, but their toxic mechanism is still a matter of debate. Here we present a combined analysis of physiology and transcriptome of Arabidopsis thaliana leaves exposure to 30 mg L-1 AgNPs and Ag+ for six days to explore the toxicity mechanism of AgNPs on Arabidopsis. Both transcriptomic and physiological results showed that AgNPs induced reactive oxygen species (ROS) accumulation and damaged photosynthesis. The toxicity of AgNPs is not merely attributable to Ag+ release and much higher photosynthetic toxicity and ROS accumulation were observed in 30 mg L-1 AgNPs than that in 0.12 mg L-1 Ag+. About 60% genes were similarly up- or down-regulated at the same concentration of AgNPs and Ag+ and these genes were enriched in photosynthesis and response to the stimulus. However, 302 genes, including those involved in glucosinolates synthesis, were specifically regulated under AgNPs treatments. In conclusion, more than the released Ag+, nanoparticle-specific effects are responsible for the toxicity of AgNPs in Arabidopsis thaliana.

Correlation of Apo-A and Apo-B/Apo-A ratio with the degree of coronary artery stenosis in patients with coronary heart disease / 西安交通大学学报(医学版)

Objective: To investigate the correlation of Apo-A and Apo-B/Apo-A ratio with the degree of coronary artery stenosis (CHD) in patients with coronary heart disease. Methods: Totally 234 patients with coronary heart disease examined with coronary angiography were collected. We collected blood lipid and calculated blood lipid ratio, such as non-HDL-C, TG/HDL-C, TC/HDL-C, LDL-C/HDL-C, and Apo-B/Apo-A. Gensini score was calculated according to the result of CAG. We analyzed the correlation between the blood lipid indicators and the Gensini score with the Spearman correlation analysis. Linear regression analysis was made of the correlation between those meaningful indicators and the Gensini score. Results: There were no significant differences in age, hypertension, diabetes or smoking index between all Gensini score groups. The illness course in middle-score group was longer than that in low-score group and high-score group (P=0.023, P=0.002). There was a significant difference in Apo-A between the groups (P=0.009, P<0.001, P=0.013). The levels of TC, Apo-B and LPα in high-score group were lower than those in low-score group (P=0.008, P=0.001, P=0.002). The levels of HDL-C and Apo-B/Apo-A ratio in both middle-score and high-score group were lower than those in low-score group (P=0.008, P=0.001). The Spearman correlation analysis between various risk factors and Gensini score found that HDL-C and Apo-A had negative correlation with Gensini score (r=-0.166, r=-0.294), the ratios of LDL-C/HDL-C and Apo-B/Apo-A were positively correlated with Gensini score (r=0.159, r=0.170). By multi-factor linear regression, Apo-A was negatively related with Gensini score (β=-62.249), and Apo-B/Apo-A ratio was positively associated with Gensini score (β =31.311). Conclusion: Apo-A and Apo-B/Apo-A ratios are the independent risk factors for the stenosis degree of coronary artery in patients with CHD. They are reliable predictors for the risk of CHD.

The prognosis value of comprehensive geriatric assessment in elder patients with acute myeloid leukemia in a single center / 中华血液学杂志

Objective: To evaluate the prognostic significance of comprehensive geriatric assessment (CGA) in Chinese elderly acute myeloid leukemia (AML) patients. Methods: 73 AML patients over the age of 60 were enrolled. CGA stratification included the following 3 instrument assessment: activity of daily living (ADL) ; instrumental activity of daily living (IADL) ; comorbidity score according to the Modified cumulative illness rating score for geriatrics (MCIRS-G) . According to CGA and age, the enrolled patients were grouped into 'fit', 'unfit' and 'frail' categories. Results: The median age of 73 elderly AML patients were 75 years old. According to CGA, 37 (50.1%) patients were classified as 'fit', 14 (19.2%) as 'unfit', and 22 (30.7%) as 'frail'. 33 (89.2%) patients in fit group received induction chemotherapy, or demethylation treatment, as 8 (57.9%) in unfit, 10 (45.5%) in frail. The overall response rate was 68.7%、62.5%, 75.0% in fit, unfit, and frail group, respectively (χ(2)=0.615, P=0.769) .The early mortality (8 weeks) in three groups were different: 5.4%, 7.1%, 27.3%, respectively (P<0.05) . The 1-year overall survival in the 'fit', 'unfit' and 'frail' groups was 64.9%, 28.6% and 22.7%, respectively (P<0.05) . The CGA score, age, ECOG score, WHO classification (2016) were the prognostic factors of AML patients. Conclusion: CGA can be used to determine the prognosis of elderly AML patients.

Advances in self-assemblied DNA nanocages as drug delivery systems / 中国药科大学学报

DNA nanocages are nanoscale three-dimensional polyhedral structures formed by self-assembled oligonucleotide strands by base pairing.DNA nanocages,especially the tetrahedron DNA nanocages,were proved to be potential nanocarriers for drug delivery,due to their good stablility,high compatibility,intracellular ablility and hollow cavity for modification and drug loading..In this paper,we reviewed the structural characteristics,synthetic methods and targeted modification of DNA nanocages,as well as their development and application in drug carriers,biological imaging,in vitro diagnosis and materials science.

Influence of prehospital emergency care connected green channel on rescue effect in patients with acute myocardial infarction / 心血管康复医学杂志

Objective:To explore rescue effect of prehospital emergency care connected green channel on patients with acute myocardial infarction (AMI).Methods:A total of 122 emergency AMI patients treated in our hospital were selected.Ac-cording to hospitalization order,they were divided into routine emergency care group (the first 61 inpatients)and green channel group (the following 61 inpatients,received prehospital emergency care connected green channel besides routine e-mergency care).Rescue effective rate,myocardial perfusion time,time of in-hospital stay and nursing satisfaction were compared between two groups.Results:Total effective rate of green channel group was significantly higher than that of routine emergency care group (78.69% vs.68.85%,P =0.028);compared with routine emergency care group,there were significant reductions in myocardial perfusion time [(100.72±10.40)min vs.(56.02±5.49)min]and time of in-hospital stay [(14.34 ± 3.46)d vs.(11.86 ± 4.24)d](P < 0.01 both),and significant rise in nursing satisfaction (81.82% vs.96.67%,P =0.028)in green channel group.Conclusion:Prehospital emergency care connected green chan-nel can improve rescue effective rate,reduce time of in-hospital stay and improve nursing satisfaction in patients with acute myocardial infarction,which is worth extending in clinic.

Correlation between HLA-DB1 genes and susceptibility to echinococcosis in Tibetan population in Tibetan Plateau / 中国血吸虫病防治杂志

Objective To determine the susceptibility genes and resistance genes in HLA-DRB1 alleles in Tibetan patients with cystic and alveolar hydatid diseases,so as to provide the references for the research of the genetic characteristics and infec-tion mechanism of Tibetan hydatid diseases. Methods The case control method was applied. The Tibetan patients with cystic and alveolar hydatid diseases(63 and 73 cases respectively)in Yushu and Guoluo Tibetan Autonomous Prefecture,and unrelat-ed healthy people(60 cases)in this area were selected as the study subjects. The polymerase chain reaction-sequence based typ-ing(PCR-SBT)technique was applied for genotyping of HLA-DRB1,and the comparison of the gene frequency. Results The frequency of HLA-DRB1*04 in the alveolar/cystic echinococcosis group was lower than that in the control group(χ2 =4.71, 4.31,both P<0.05). Conclusion HLA-DRB1*04 genotypes may be associated with the resistance of cystic and alveolar echi-nococcosis and its resistance genes.

Influence of prehospital emergency care connected green channel on rescue effect in patients with acute myocardial infarction / 心血管康复医学杂志

Objective:To explore rescue effect of prehospital emergency care connected green channel on patients with acute myocardial infarction (AMI).Methods:A total of 122 emergency AMI patients treated in our hospital were selected.Ac-cording to hospitalization order,they were divided into routine emergency care group (the first 61 inpatients)and green channel group (the following 61 inpatients,received prehospital emergency care connected green channel besides routine e-mergency care).Rescue effective rate,myocardial perfusion time,time of in-hospital stay and nursing satisfaction were compared between two groups.Results:Total effective rate of green channel group was significantly higher than that of routine emergency care group (78.69% vs.68.85%,P =0.028);compared with routine emergency care group,there were significant reductions in myocardial perfusion time [(100.72±10.40)min vs.(56.02±5.49)min]and time of in-hospital stay [(14.34 ± 3.46)d vs.(11.86 ± 4.24)d](P < 0.01 both),and significant rise in nursing satisfaction (81.82% vs.96.67%,P =0.028)in green channel group.Conclusion:Prehospital emergency care connected green chan-nel can improve rescue effective rate,reduce time of in-hospital stay and improve nursing satisfaction in patients with acute myocardial infarction,which is worth extending in clinic.

Synthesis and evaluation of effective photoaffinity probe molecule of furospinosulin-1, a hypoxia-selective growth inhibitor.

The synthesis and evaluation of a photoaffinity probe molecule for furospinosulin-1, a hypoxia-selective growth inhibitor that we identified from marine sponge, was studied. An analogue carrying an alkyne tail showed potent hypoxia-selective inhibitory activity exceeding that of the parent molecule, and exhibited in vivo anti-tumor activity following oral administration. The alkyne moiety in the analogue was also found to be a good anchoring group for the preparation of probe molecules; a photoaffinity probe molecule having an optimized spacer length was selected through the systematic synthesis of several probes and the evaluation of their hypoxia-selective growth inhibitory activity and electrophoretic mobility shift properties.

Identification of two severe fever with thrombocytopenia syndrome virus strains originating from reassortment.

Recently, a novel bunyavirus, severe fever with thrombocytopenia syndrome virus (SFTSV), was isolated in central China. The virus can cause multi-clinical symptoms: severe fever, thrombocytopenia, leukocytopenia, with a mortality rate of ~10%. Several studies show that SFTSV could undergo rapid evolution via gene mutation and homologous recombination. However, as an important evolutionary force for segmented-genome viruses, reassortment has not been reported in SFTSV. In this study, we identified two SFTSV strains of which the S segment has different origin from M and L, suggesting that reassortment might be potential force driving rapid change of SFTSV. This result might shed new light on the evolutionary behavior of the novel virus.

Probe molecule equipped with boronic acid moiety as a reversible cross-linking group improves its binding affinity.

Syntheses of biotinylated probe molecules of L-glutathione (GSH) equipped with boronic acid moiety and evaluation of their binding affinities against glutathione-S-transferase (GST) were described. It revealed that the presence of boronic acid moiety in an appropriate position enhances binding affinity of GSH probe toward GST probably by forming a reversible cross-link. Among prepared, the boronate-containing probe 8b exhibited the highest recovering ability of GST from Escherichia coli cell lysate.

Quantification of GRAF gene expression in patients with acute myeloid leukemia using EvaGreen real time quantitative PCR / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To quantify the expression level of GRAF gene in acute myeloid leukemia (AML) and analyze its clinical significance.</p><p><b>METHODS</b>The EvaGreen real-time quantitative polymerase chain reaction (RQ-PCR) assay was established and performed to measure the GRAF gene transcripts in 71 cases with AML and 21 with nonmalignant hematological diseases. The clinical correlation of GRAF expression was analyzed.</p><p><b>RESULTS</b>The established EvaGreen RQ-PCR assay had good specificity, reproducibility and sensitivity. The GRAF expression level was significantly lower in AML (0.01%-169.75%, median 3.82%) than that in controls (14.49%-126.85%, median 56.04%) (P<0.05). There was no correlation between the level of GRAF transcript and the sex, age, hematologic parameters, FAB subtypes and karyotypic groups (P>0.05).</p><p><b>CONCLUSION</b>The GRAF gene was down-regulated in AML, which might play a role in the leukemogenesis.</p>

Quantification of the PRAME transcripts in patients with acute myeloid leukemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the expression level and clinical significance of the preferentially expressed antigen of melanoma (PRAME) transcripts in patients with acute myeloid leukemia (AML).</p><p><b>METHODS</b>Real-time quantitative polymerase chain reaction with EvaGreen dye was established to detect the expression level of PRAME transcripts in the bone marrow mononuclear cells of 56 AML cases and 20 controls. The clinical association of PRAME transcripts was analyzed.</p><p><b>RESULTS</b>The PRAME transcripts were 0-1.46% (median 0.18%) and 0-21 618.09% (median 9.79%) in controls and AML cases, respectively (P< 0.01). Among the FAB subtypes, those with M1, M2, M3 and M4 had significantly higher level of PRAME transcripts than controls. However, those with M5 had similar level of PRAME transcripts as controls. There was a significantly negative correlation between the PRAME transcripts and cytogenetic risk groups (r= -0.438, P= 0.001). Cases in low risk had significantly higher level of PRAME transcripts than those in intermediate and high risk. Among cases with AML-M2, those with t(8;21) had significantly higher level of PRAME transcripts (135.06% -21 618.09%, median 2201.88%) than those without t(8;21)(0.14% -1696.30%, median 17.97%)(P= 0.002). In a patient with sequential samples, PRAME transcripts significantly decreased after induction therapy and significantly increased after relapse.</p><p><b>CONCLUSION</b>The PRAME transcript was highly expressed in AML patients and was a favorable marker of prognosis. Quantification of PRAME transcript can be used in monitoring disease status of AML.</p>

Expression of PML-RAR alpha fusion transcripts detection by using RQ-PCR / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish the real time quantitative PCR (RQ-PCR) assay according to 'Europe Against Cancer' (EAC) program and analyze the results of detection and quantification of different PML-RAR alpha transcript isoforms in patients with acute promyelocytic leukemia (APL).</p><p><b>METHODS</b>Three RQ-PCR systems were performed to detect the most frequent PML-RAR alpha transcripts (L-form, S-form and V-form) in 30 APL patients and the RQ-PCR end-point products were identified by electrophoresis.</p><p><b>RESULTS</b>S-form RQ-PCR system could amplify positive signals of three isoforms in all of 30 cases, and V-form RQ-PCR system could do so in both L-form and V-form positive cases, however, L-form RQ-PCR system could only do so in L-form-positive cases. Electrophoresis and sequencing of end-point products amplified by S-form RQ-PCR system revealed three bands in each of L-form (621 bp, 477 bp and 218 bp) and V-form (567 bp, 423 bp and 218 bp) positive patients samples.</p><p><b>CONCLUSIONS</b>RQ-PCR, sensitive and reliable, can be used for monitoring the minimal residual disease in APL patients, however, its results should be interpreted carefully if it is used for detection of PML-RAR alpha fusion transcripts prospectively.</p>

Quantification of PML/RAR alpha fusion gene transcripts in patients with acute promyelocytic leukemia by using real-time quantitative PCR / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish and evaluate a real time quantitative PCR (RQ-PCR) method for detection and quantification the PML/RAR alpha fusion gene transcripts in patients with acute promyelocytic leukemia (APL).</p><p><b>METHODS</b>Three pairs of primers and TaqMan probe were designed for detecting the most frequent PML/RAR alpha transcripts (L-form, S-form and V-form) and normal ABL was used as an internal control. A real time PCR condition was established to detect PML/RAR alpha and ABL positive templates with a series of dilutions. To evaluate this assay, bone marrow samples from 6 APL patients were detected.</p><p><b>RESULTS</b>In repeated tests, maximal sensitivities of 10 copies/microL were obtained, while reproducible maximal sensitivity achieved 100 copies/microL. In 10 normal controls, no amplified fluorescent signals were detected. The median absolute and normalized amount of PML/RAR alpha fusion gene transcripts were 4.27 x 10(3)-3.36 x 10(5) copies/50 ng (median 4.33 x 10(4)copies/50 ng) and 29.38%-600.53% (median 48.12%) respectively. One case showed significant decrease of PML/RAR alpha fusion gene transcripts after induction therapy compared to that at the time of diagnosis, while the fusion transcripts significantly increased after relapsed.</p><p><b>CONCLUSION</b>RQ-PCR is a sensitive, reliable quantitative assay and can be used in the diagnosis of APL and measurement of MRD.</p>

Alteration of methylation status of fragile histidine triad gene promoter in patients with myelodysplastic syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the methylation status of fragile histidine triad (FHIT) gene promoter in patients with myelodysplastic syndrome (MDS) and its clinical relevance.</p><p><b>METHODS</b>Methylation-specific PCR (MSP) was used to detect FHIT promoter methylation in bone marrow samples from 54 MDS cases.</p><p><b>RESULTS</b>Hypermethylation of FHIT promoter was detected in 26 cases (48.1%). Association was not found between FHIT gene hypermethylation and sex, hematologic parameters and chromosomal abnormalities of MDS patients, but found between FHIT gene hypermethylation and age of the MDS cases. Although significant difference was not observed in the frequencies of FHIT gene hypermethylation among patients with refractory anemia/refractory anemia with ringed sideroblasts (RA/RAS) (1/6, 16.7%), refractory anemia/refractory anemia with ringed sideroblasts (RCMD) and refractory cytopenia with multilineage dysplasia with ringed blasts (RCMD-RS) (6/19, 31.6%), refractory anemia with excess blasts-1 (RAEB-1) (7/11, 63.6%), refractory anemia with excess blasts-2 (RAEB-2) (4/7, 57.1%) and refractory anemia with excess blasts in transformation/acute myeloid leukemia (RAEBt/AML) (8/11, 72.7%)(chi-square=8.417, P=0.077), it was observed in patients in early stages (RA/RAS and RCMD) (7/25, 28.0%), advanced stages (RAEB-1 and RAEB-2)(11/18, 61.1%) and RAEBt/AML (8/11, 72.7%) (chi-square=7.938, P=0.019). Furthermore, there was a positive correlation between the frequency of FHIT gene hypermethylation and different IPSS groups (chi-square=10.110, P=0.018).</p><p><b>CONCLUSION</b>FHIT gene hypermethylation might be one of the molecular events involved in the disease progression of MDS.</p>

A new eudesmane sesquiterpene lactone from Curcuma wenyujin.

The aim of the study was to investigate the sesquiterpene constituents from the rhizomes of Curcuma wenyujin Y. H. Chen et C. Ling. The isolation and purification of the constituents from the 50% EtOH extracts of the rhizomes were performed with repeated column chromatography over sillica gel and macroporous resin. Eight sesquiterpenes were obtained and identified as wenyujinlactone A (1), neolitamone A (2), zedoarondiol (3), isozedoarondiol (4), aerugidiol (5), curcumol (6), curdione (7) and (1R, 10R)-epoxy-(-)-1, 10-dihydrocurdine (8) by means of spectral analysis. Among them, compound 1 was found to be a new eudesmane sesquiterpene lactone, whilst compounds 2-5 were obtained from this plant for the first time.