Objective:To investigate the effects of clinically achievable dose of lovastatin on prostate cancer PC3 cells.Methods:PC3 prostate cancer cells were treated with dimethyl sulfoxide(DMSO),or lovastain only,or lovastatin with mevalonic acid for 24,48 and 72 hours respectively.MTT assay was used to detect the cell viability.By means of [3H] thymidine incorporation tests,the effects of lovastatin on cell proliferation were analyzed.Western blot was used to detect activated casepase3,caspase7,and cleaved PARP(cPARP),the important molecules on the apoptosis pathway.Results:Cell proliferation of PC3 was significantly inhibited by 39.29%[(63.69%?3.69%) vs(102.98%?6.84%),P=0.000] after 48 h treatment with lovastatin at its clinically achievable dose of 2 ?mol/L.After 72 hours the cell proliferation was inhibited by 44.24% [(52.79%?9.88%) vs(97.03%?0.87%),P=0.048].The cell number was also markedly decreased(4.86?105 ? 0.10?105) vs(9.66?105?0.10?105),P=0.000] after 72 h treatment at this low concentration of 2 ?mol/L.The viability of PC3 cells was significantly decreased 50.12%(56.52%?6.40%) vs(106.64%?5.27%),P=0.000] and 60.05%(41.99%?11.64%) vs(102.94%?8.49%),P=0.000] after 48 h and 72 h treatment,respectively.In addition,2 ?mol/L lovastatin induced activation of casepase7 and led the death substrate PARP to cleavage.Conclusion:Clinically achievable dose of lovastatin inhibits prostate cancer PC3 cell proliferation and induces PC3 cell apoptosis.

Objective To construct and identify the efficacy of a lentiviral vector harboring RNAi sequence targe-ting NSBP1 gene. Methods Three siRNA targeting the NSBP1 mRNA were designed, the pGCSIL-GFP-NSBP1 lentivirus vectors were constructed and confirmed by DNA sequencing. A total of 293T cells were co-transfected with pGCSIL-GFP-NSBP1, pHelper1.0 and pHelper2.0 for the virus stocks produced, the titer of the virus was test-ed. After lentivirus transfecting into DU145 ceils, Western-blot and MTT methods were used to determine the ex-pression and biological activity of NSBP1 gene, the cells were transplanted into nude mice, then inhibitive effect was observed. Results PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting the hu-man NSBP1 gene was successfully inserted into the lentiviral vector. The titer of the recombinant]entiviral vector was 2 × 10~8TU/mL. NSBP1 protein expression level in transfected cells was significantly decreased and growth rate of cells transfected with lentivirus was decreased by MTT assay, the downregulation of NSBP1 reduced growth rate of transplantated tumor, whereas tumorgenicity was not influenced. Conclusion The construction of the]entiviral vector of NSBP1 has been successfully prepared and NSBP1 plays an important regulatory role in androgen-inde-pendent prostate cancer cell proliferation.

OBJECTIVETo study the effect of 5-fluorouracil-FU in combination with astragalus membranaceus(AM) on amino acid metabolism in mice model of gastric carcinoma induced by 3-methylcholanthrene(MC).

METHODSMice gastric carcinoma models were established by 3-methylcholanthrene induction and randomly divided into different groups, and received 5-FU treatment (group A) 5-FU plus AM (group B), 5-FU plus a high dose of AM(group C), no treatment (group D). Normal mice were used as control (group N). Free amino acid in the tumor specimens were examined.

RESULTSThe levels of free Valine, Methionine, Leucine, Arginine and cystine in the tumor specimens in group D were significantly higher than that in group N(P< 0.05). The levels of free serine in group A, B, C, D were significantly higher than that in group N. The levels of free glutamic acid in group A, B were significantly higher than that in group N(P< 0.05). The levels of free proline in group C, D were significantly higher than that in group P, N(P< 0.05).

CONCLUSIONSThe increasing levels of free serine and proline in tumor specimens in gastric cancer mice model reveals metabolic disturbance of amino acid. 5-FU plus astragalus membranaceus can decrease the level of free glutamic acid in the mice models, and inhibit tumor growth.

Amino Acids ; metabolism ; Animals ; Astragalus membranaceus ; Fluorouracil ; therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Phytotherapy ; Stomach Neoplasms ; drug therapy ; metabolism

Objective To study the effects of Xuebijing injection,somatostatin monotherapy and their combination therapy on the improvement of inflammation and prognosis in acute severe pancreatitis.Methods A total of 90 patients with acute severe pancreatitis treated in our hospital were enrolled in this study.Patients were divided into group A,group B and group C,with 30 cases in each group.The patients in group A were treated with Xuebijing injection alone,and the patients in group B were treated with somatostatin alone,and patients in group C were treated with Xuebijing injection and somatostatin,and all the treatment was lasted for 7 days.The alleviation time of clinical symptoms,improvement of inflammation and prognosis in three groups before and after treatment were observed and compared.Results After treatment,the anal defecation recovery time,abdominal pain relief time,bowel sounds recovery time and abdominal distension time in the three groups were lower than treatment before,and group C were lower than group A and group B (P ＜ 0.05).The levels of interleukin-6 (IL-6),tumour necrosis factor-α (TNF-α) and C-reactive protein (CRP) were significantly lower in group C than those in group A and group B (P ＜0.05).The APACHE-Ⅱ scores after treatment was higher than treatment before,and C group was lower than A group,B group (P ＜0.05).Conclusion Xuebijing injection in combined with somatostatin for acute severe pancreatitis has significantly better efficacy than monotherapy,and it can improve the efficacy of inflammation treatment,and prognosis.

Objective To study the effects of Xuebijing injection,somatostatin monotherapy and their combination therapy on the improvement of inflammation and prognosis in acute severe pancreatitis.Methods A total of 90 patients with acute severe pancreatitis treated in our hospital were enrolled in this study.Patients were divided into group A,group B and group C,with 30 cases in each group.The patients in group A were treated with Xuebijing injection alone,and the patients in group B were treated with somatostatin alone,and patients in group C were treated with Xuebijing injection and somatostatin,and all the treatment was lasted for 7 days.The alleviation time of clinical symptoms,improvement of inflammation and prognosis in three groups before and after treatment were observed and compared.Results After treatment,the anal defecation recovery time,abdominal pain relief time,bowel sounds recovery time and abdominal distension time in the three groups were lower than treatment before,and group C were lower than group A and group B (P ＜ 0.05).The levels of interleukin-6 (IL-6),tumour necrosis factor-α (TNF-α) and C-reactive protein (CRP) were significantly lower in group C than those in group A and group B (P ＜0.05).The APACHE-Ⅱ scores after treatment was higher than treatment before,and C group was lower than A group,B group (P ＜0.05).Conclusion Xuebijing injection in combined with somatostatin for acute severe pancreatitis has significantly better efficacy than monotherapy,and it can improve the efficacy of inflammation treatment,and prognosis.

To explore the difference of biological characteristics between two subpopulations of adult bone marrow mesenchymal stem cells (MSC), this study was designed to observe the morphological feature and immunophenotype of the adult MSC in the ex vivo culture, the mononuclear cells isolated from normal adult bone marrow were cultured in DMEM with 10% fetal bovine serum. Cell morphology, immunophenotype and cell cycle of two different subgroups were investigated. Cells from 80% confluence were passed through a 10 microm filter, then the fillered cells were cultured in the semisolid methylcellulose medium. The results showed that (1) two different subpopulations were observed in the ex vivo culture. The fibro-like cell was called mature MSC (mMSC) and the smaller round cell was defined rapidly as MSC self-renewing cells (RS cells); (2) the average proportion of cells in G(0)/G(1) of RS cells was approximately 99%, but that of mMSCs was 90%; (3) both of the two populations were negative on the lineage-committed antigen (such as CD34, CD45, CD3, CD19, CD33, HLA-DR, CD38), while positive on the expression of CD90, CD105, C166, CD29, CD44, CD49e, CD54, CD13. However, the expression of these antigens on RS cells was weaker than that on mMSC, but CD117 and KDR were higher expressed when compared with the mMSC; (4) after 4 to 5 week semisolid culture, no hematopoietic progenitor cell colonies were observed. It is concluded that adult MSCs are heterogeneous in that distinct morphological populations exist. The RS cells appear to be the more primitive with greater potential for self-renewal and multilineage differentiation.
Adult ; Antigens, CD ; analysis ; Bone Marrow Cells ; cytology ; immunology ; Cell Cycle ; Cell Differentiation ; immunology ; Cell Lineage ; immunology ; Cell Size ; Cells, Cultured ; Humans ; Immunophenotyping ; Leukocytes, Mononuclear ; cytology ; immunology ; Mesenchymal Stromal Cells ; cytology ; immunology

To investigate the influence of G-CSF mobilization on functions of donor T lymphocyte subpopulation and acute graft-versus-host disease, peripheral blood samples of 20 healthy donors were collected before and after G-CSF mobilization. The whole blood was diluted with IMDM in ratio of 1:1 and then incubated with PMA + ionomycin + monensin at 37 degrees C, 5% CO2 for 4 hours. After being mobilized and stained, the IL-4, IFN-gamma and IL-2 positive cells were counted with three-color flow cytometry. The results showed that before G-CSF mobilization, the percentages of donor's CD3(+)IFN-gamma(+), CD4(+)IFN-gamma(+), CD8(+)IFN-gamma(+) T cells were 3.2% (0% - 45.9%), 1.3% (0% - 23.8%) and 1.5% (0% - 22.2%) respectively. The percentage of above mentioned cells in donor increased to 19.2% (0% - 53.9%), 9.5% (0% - 49.5%), 7.5% (0% - 38.1%) respectively after G-CSF mobilization. The IL-2 positive CD3(+), CD4(+) and CD8(+) T cell percentage in pre-G-CSF mobilized donors was 1.5% (0% - 31%), 0.8% (0% - 30.0%) and 0% (0% - 5.3%) respectively and subsequently increased to 25.7% (0% - 51%), 19.8% (0% - 39.7%), 4.6% (0% - 20.9%) respectively after G-CSF mobilization. The IL-4 positive T subpopulation did not increased significantly after G-CSF mobilization. In the early stage after peripheral blood stem cell transplantation, donor's Tc1 percentage in aGVHD group was significantly higher than that in non-aGVHD group. The morbidity of severe aGVHD in high Tc2 percentage group was significantly lower than that in low Tc2 percentage group. It is concluded that the donor's type I T cells increase after G-CSF mobilization, the Tc1 percentage of G-CSF mobilized donor is correlated with the occurrence of aGVHD in the early stage after HSCT, the percentage of Tc2 in donor is negatively correlated with aGVHD morbidity in recipients.
Adolescent ; Adult ; Female ; Graft vs Host Disease ; etiology ; Granulocyte Colony-Stimulating Factor ; adverse effects ; Hematopoietic Stem Cell Mobilization ; adverse effects ; methods ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; therapy ; Leukemia, Myeloid, Acute ; therapy ; Male ; Middle Aged ; Recombinant Proteins ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes ; immunology

Within the past few years, the invention of next-generation sequencing has revealed several new genes associated with tumor formation and development, for example DNMT3a. This gene is an independent prognostic factor for acute myeloid leukemia (AML). The objective of this study was to analyze the DNMT3a mutation in childhood AML in a single center. PCR amplification of the entire coding region of DNMT3a was performed using 23 overlapping primer pairs in 57 patients who were diagnosed in Blood Disease Hospital of Chinese Academy of Medical Sciences, then the directly sequencing was underwent. The results showed that no DNMT3a mutation was found in these patients including the hotspot R882. But AML1/ETO mutation was found in 10 patients, CBFB/MYH11 mutation in 3 patients, PML/RARa mutation in 13 patients, FLT3/ITD mutation in 5 patients, FLT3/TKD mutation in 1 patient, PML/RARa and FLT3/TKD mutation coexisted in 2 patients. It is concluded that DNMT3a mutations are rare in childhood AML, and different mechanisms of myeloid leukemogenesis between childhood and adults maybe involved.
Adolescent ; Base Sequence ; Child ; Child, Preschool ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Infant ; Karyotyping ; Leukemia, Myeloid, Acute ; genetics ; Male ; Mutation

Objective:To explore sex and rural-urban differences in the associations of different blood pressure levels with the risk of prediabetes.Methods:We used a multi-stage stratified cluster random sampling method to investigate 21 637 residents aged ≥18 years from 10 survey areas in Hubei province in 2020. The data on questionnaire, physical measurements, and laboratory indicators of the participants were collected. The associations of different blood pressure levels with risk of prediabetes by sex and regions were analyzed using multivariate logistic regressions after complex weighting.Results:A total of 16 111 subjects were included. The prevalence (95% CI) of prediabetes, impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and IFG complicated with IGT were 25.1% (14.4%-35.9%), 12.7% (3.2%-22.1%), 8.1% (6.3%-9.8%), and 4.4% (2.3%-6.5%), respectively. After multivariate adjustment, the risk of prediabetes, IFG, IGT, and IFG complicated with IGT increased with the increment of blood pressure (both P for trend <0.05). The positive dose-response relationships between blood pressure levels and risk of prediabetes were also significant among male, urban, and rural residents (both P for trend <0.05), and the interactions between sex and blood pressure showed significant associations for risk of prediabetes and IGT (both P for interaction <0.05). Conclusions:Higher blood pressure levels were associated with an increased risk of prediabetes. The association with prediabetes was stronger in males, but no significant difference was found between urban and rural residents. More distinctive and effective prevention and control strategies should be developed for different populations.

Objective:To evaluate the health benefits and intervention efficiency of different strategies of initiating antihypertensive therapy for the primary prevention of cardiovascular diseases in a community-based Chinese population from the Chinese electronic health records research in Yinzhou(CHERRY)study.Methods:A decision-analytic Markov model was used to simulate and compare different antihy-pertensive initiation strategies,including:Strategy 1,initiation of antihypertensive therapy for Chinese adults with systolic blood pressure(SBP)≥140 mmHg(2020 Chinese guideline on the primary preven-tion of cardiovascular diseases);Strategy 2,initiation of antihypertensive therapy for Chinese adults with SBP≥130 mmHg;Strategy 3,initiation of antihypertensive therapy for Chinese adults with SBP ≥140 mmHg,or with SBP between 130 and 140 mmHg and at high risk of cardiovascular diseases(2017 American College of Cardiology/American Heart Association guideline for the prevention,detection,evaluation,and management of high blood pressure in adults);Strategy 4,initiation of antihypertensive therapy for Chinese adults with SBP≥ 160 mmHg,or with SBP between 140 and 160 mmHg and at high risk of car-diovascular diseases(2019 United Kingdom National Institute for Health and Care Excellence guideline for the hypertension in adults:Diagnosis and management).The high 10-year cardiovascular risk was de-fined as the predicted risk over 10％based on the 2019 World Health Organization cardiovascular disease risk charts.Different strategies were simulated by the Markov model for ten years(cycles),with parame-ters mainly from the CHERRY study or published literature.After ten cycles of simulation,the numbers of quality-adjusted life years(QALY),cardiovascular events and all-cause deaths were calculated to evaluate the health benefits of each strategy,and the numbers needed to treat(NNT)for each cardiovas-cular event or all-cause death could be prevented were calculated to assess the intervention efficiency.One-way sensitivity analysis on the uncertainty of incidence rates of cardiovascular disease and probabilis-tic sensitivity analysis on the uncertainty of hazard ratios of interventions were conducted.Results:A to-tal of 213 987 Chinese adults aged 35-79 years without cardiovascular diseases were included.Com-pared with strategy 1,the number of cardiovascular events that could be prevented in strategy 2 increased by 666(95％UI:334-975),while the NNT per cardiovascular event prevented increased by 10(95％UI:7-20).In contrast to strategy 1,the number of cardiovascular events that could be prevented in strategy 3 increased by 388(95％UI:194-569),and the NNT per cardiovascular event prevented decreased by 6(95％UI:4-12),suggesting that strategy 3 had better health benefits and intervention efficiency.Compared to strategy 1,although the number of cardiovascular events that could be prevented decreased by 193(95％UI:98-281)in strategy 4,the NNT per cardiovascular event prevented decreased by 18(95％UI:13-37)with better efficiency.The results were consistent in the sensitivity analyses.Conclusion:When initiating antihypertensive therapy in an economically developed area of China,the strategy combined with cardiovascular risk assessment is more efficient than those purely based on the SBP threshold.The cardiovascular risk assessment strategy with different SBP thresholds is suggested to balance health benefits and intervention efficiency in diverse populations.