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Objective@#Study on the improvement of quality standard for Zicao ointment.@*Methods@#The content of shikonin and β,β’-dimethylacrylshikonin was determined by high performance liquid chromatogram (HPLC). The column was Inertsil ODS-3 C18 column (4.6 mm×250 mm, 5 μm). The mobile phase comprising of acetonitrile-0.05% formic acid solution (70:30). The flow rate was 1.0 ml/min, the detection wavelength was 275 nm, and the column temperature was 25 ℃.@*Results@#The linear range of shikonin and β,β’-dimethylacrylshikonin were 0.04-0.81 μg (r=0.999 5) and 0.41-10.36 μg (r=0.999 2). The average recovery were 103.32% and 101.80%, the RSD were 1.96% and 2.11%.@*Conclusions@#The method is simple, accurate and sensitive to control the quality of Zicao ointment.
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Objective To investigate the effects of long-term consumption of Fallopia multiflora on mouse hematopoietic system.Methods Forty 10-month old female C57BL/6J mice were equally divided into two groups at random, the control group fed with normal food , and the experimental group , given food with added Fallopia multiflora. After 10 month, the mice were sacrificed, and the peripheral blood, spleen, thymus and bone marrow cells were examined by flow cytometry.Results In the mice fed with Fallopia multiflora, the percentage of B cells in the spleen and CD 4 +cells in the thymus were increased , and CD8 + cells in the thymus and bone marrow hematopoietic stem cells were decreased , among the bone marrow cells , G0 cells were increased , but G1 and G2/S/M cells decreased .Conclusions Long-term proper consumption of Fallopia multiflora can delay the ageing of the hematopoietic system , and sustain its stability.
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Objective To investigate the influence of Siraitia Grosvenori and Rehmannia Glutinosa on the Hematopoietic stem cells proliferation and function .Methods Cells from the peripheral blood , spleen and bone marrow of mice were stained with indicated antibodies , and analyzed by flow cytometry .Mice were divided 3 groups:control group, Siraitia Grosvenori treatment group and Rehmannia Glutinosa treatment group .After 4.5Gy IR treatment, mice divided 4 groups: control group, 4.5Gy IR treatment and feed with normal food, 4.5Gy IR treatment and feed with Siraitia Grosvenori and 4.5Gy IR treatment and feed with Rehmannia Glutinosa for 1 month.Results Mice fed with Siraitia Grosvenori and Rehmannia Glutinosa decreased the percentage of B cells and increased the percentage of M cell .For HSCs, the number of HSCs was increased , especially the number of LT-HSCs.After 4.5Gy IR treatment, mice fed with Siraitia Grosvenori and Rehmannia Glutinosa increase the number of HSCs , and increased the percentage of M cells . Conclusion Siraitia Grosvenori and Rehmannia Glutinosa promote the hematopoietic stem cells and progenitor cells proliferation and function and recover the damage that caused by IR treatment .
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Objective To study the influence of IL-33 on the Hematopoietic stem cells and progenitor cells . Methods Cells from the peripheral blood , spleen, thymus and bone marrow were stained with indicated antibodies and analyzed by flow cytometry . The LT-HSCs were sorted and culture using in vitro clonogenic assay . Results The percentage of B cells and T cells was decreased and the percentage of M cells was increased in the peripheral blood from IL -33 transgenic mice .Compared with the wildtype mice , the number of HSCs , MPPs and CLP was decreased;meanwhile the number of CMP and GMP was increased in the bone marrow from IL-33 transgenic mice .An in vitro clonogenic assay showed that LT-HSCs increased the ability to self-renew from IL-33 transgenic mice .And the percentage of S-G2-M stage hematopoietic stem cell was increased from IL-33 transgenic mice .Conclusion IL-33 increase the myeloid differentiation in hematopoietic stem cells .
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Objective Systematic review the effect and side effect of Probucol on contrast-induced acute kidney injury (CIAKI), and to evaluate the characteristics and strength of the protective effect. Methods Electronic search studies from databases published in English and Chinese before March 2014, and hand searches of relevant randomized controlled trials of references. Features, the quality of research and valid data of included studies were extracted, then using RevMan 5.0 software to conduct a Meta-analysis base on Cochrane systematic review methods. Results A total of 11 literatures described eight randomized controlled trials involving 1938 cases of patients evaluated the effect of Probucal on CIAKI. A total of seven studies involving 1,298 patients, compared the incidence of CIAKI, the total incidence rate was 10.9%(141/1298), CIAKI incidence was 5.7%(37/652) in the Probucal group and 16.1%(104/646) in the control group (RR 0.37, 95%CI 0.26~0.53). Lower postoperative creatinine values was observed in Probucol group, compared with the control group on the first day Weighted Mean Difference (WMD-6.76, 95%CI-9.33~-4.20)μmol/L, the second day (WMD-16.90, 95%CI-22.61~-11.19)μmol/L, the third day(WMD-11.05, 95%CI-17.65~-4.45)μmol/L, and lower peak postoperative creatinine than the control group[(WMD- 14.58, 95%CI- 19.00 ~ - 10.16)μmol/L]. Probucol group with lower postoperative urinary KIM-1[(WMD-3.64 , 95%CI-3.72~-3.57) ng/ml], and the serum CysC was also low. Conclusions Preoperative or postoperative oral Probucol has a protective effect on CIAKI, can reduce the rise of postoperative renal injury indicators of coronary angiography or coronary intervention, such as serum creatinine and CysC, and CIAKI can be reduced by about 60%, no significant side effects observed.
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Objective To study the influence of Apo E gene knockout on the lymphocytes . Methods Cells from the peripheral blood, spleen and bone marrow of Apo E knockout and wildtype mice were stained with kinds of antibodies , and analyzed by flow cytometry .Results Compared with wildtype mice , significant differences were found in B and T lymphoctes in the peripheral blood and spleen , but there was no significant difference in pre B cells , T lymphocytes in the thymus and long term hematopoietic stem cell in the Apo E knockout mice .Conclusion Numbers of B lymphocytes decreased in the peripheral blood and spleen , but there was no significant difference in B , T lymphocytes development , and numbers of long term hematopoietic stem cell in Apo E gene knockout mice .
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Objective In order to establish a rhesus monkey model of p53 gene silencing, firstly we screened and determined the effective silencing targets of p53 gene at the cellular level in rhesus monkey.Methods The expression of p53 gene was detected in COS-7 cells ( derived from the kidney of the African Green Monkey, Cercopithecus aethiops).Three small hairpin RNA ( shRNA) sequences targeting rhesus monkey p53 gene were designed, analysed by bioinformatics, and inserted into lentivirus-based gene silencing constructs FUGW-TDT.The plasmids of p53-RNAi and control vector were transfected into the COS-7 cells, respectively.The suppression of p53 mRNA was detected by real-time PCR, and the changes of p53 protein expression were detected by Western blot assay.Results p53 gene expression was detected in COS-7 cells.Bioinformatics analysis showed that three gene-silencing sequences were screened which lied in the open reading frame ( ORF) region and targeted 238 -258bp, 681 -701bp, 169 -189bp of the rhesus monkey p53 mRNA.At 48 hrs after transfection of the three silencing constructs, p53 mRNA was suppressed by(87.17 ±4.03)%, ( 72.62 ±4.11)% and(76.22 ±0.98 )%, and p53 protein was suppressed by ( 84.44 ±2.18 )%, ( 71.04 ±1.18)% and ( 74.17 ±0.95 )%, respectively. Conclusions We obtained three effective target sequences showing high efficiency in p53silencing, which can be used in further studies on gene silencing in rhesus monkey.
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Objective To investigate the effects of ginsenoside Rg3 on growth and apoptosis of gastric cancer cell line MKN-45 and SGC-7901 in vitro. Methods MKN-45 and SGC-7901 cells at logarithmic growth phase were obtained, and were cultured with ginsenoside Rg3 of different concentrations (20, 30, 40, 50 μg/mL) for 24, 48 h or 24, 48 and 72 h. Cells cultured without ginsenoside Rg3 were served as controls. The inhibition rates of ginsenoside Rg3 on MKN-45 and SGC-7901 cells were detected by MTT assay, apoptosis rate of SGC-7901 cells was determined by Annexin V/PI double staining flow cytometry, cell cycles of SGC-7901 cells were analysed by flow cytometry, and morphological changes of SGC-7901 cells in 50 μg/mL ginsenoside Rg3 treatment group were observed by transmission electron microscopy. Results The inhibition rates on MKN-45 and SGC-7901 cells in each ginsenoside Rg3 treatment group were significantly higher than those in control group (P < 0.05), and the inhibition rates increased with the concentrations of ginsenoside Rg3 and time of culture ( P < 0.05). Compared with control group, the apoptosis rates of SGC-7901 cells and percentages of cells in G_1/G_1 cell cycle in each ginsenoside Rg3 treatment group were significantly increased in a concentration and time dependent manner. Typical morphology of SGC-7901 cell apoptosis was observed by transmission electron microscopy in 50 μg/mL ginsenoside Rg3 treatment group. Conclusion Ginsenoside Rg3 has significant inhibition effect on gastric cancer cell lines in vitro with a concentration and time dependent manner, the mechanism of which may involve the induction of gastric cell line apoptosis.
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Objective To study the changes in the percentage of CD+4 CD+25 regulatory T (Tr) cells in peripheral blood and deciduas in unexplained recurrent spontaneous abortion (URSA) patients, normal non-pregnant and pregnant women respectively. Methods The percentage of CD+4 CD+25 Tr cells in deciduas and peripheral blood from 25 URSA patients, 22 normal non-pregnant (NNP) women, and 34 normal early pregnant (NP) women were measured by double-staining followed by flow cytometric analysis. Results (1) The percentage of CD+4 CDbright25 T cells in peripheral blood in both URSA and NP [ ( 1.55±0.77 ) %, (2. 65±1.10)%, respectively] women were increased significantly than that in NNP women [ (0. 39± 0.14)% P<0.05]. The percentage of CD+4 CDbright25 T cells in peripheral blood in URSA women was significantly lower than that in NP women (P<0.05 ). (2) The percentage of CD+4 CDbright25 T cells in decidua in URSA women was significantly lower than that in NP women [ ( 0. 59±0. 23 ) %, ( 1.24 ± 0. 55)%, respectively, P <0. 01 ]. There was no significant difference in the percentage of CD+4 CDdim25 T cells in decidua between URSA women and NP women [ (4. 23±1.52)%, (3.75±1.88)%, respectively, P>0.05]. (3) The proportion of CD+4 CDbright25/CD+4 cells in deeiduas was significantly higher than that in peripheral blood in NP women [(13. 10±10. 25 ) %, ( 5.59±2.62 ) %, respectively, P < 0. 05 ] . However, a significant difference in the proportion of CD+4 CDbright25/CD+4 between decidua and peripheral blood was not found in URSA patients [ (5. 16±2. 83 ) %, ( 4. 64±2. 07 ) %, respectively, P>0.05)].Conclusions The number of CD+4 CD+25 Tr cells is increased in normal pregnancy and decreased in URSA. Therefore, CD+4 CD+25 Tr cells may play an important role in maintaining maternal-fetal tolerance and may be involved in the pathogenesis of URSA.
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<p><b>OBJECTIVE</b>To investigate whether ascorbic acid could enhance the efficacy of arsenic trioxide (As(2)O(3)) combined with 2, 3-dimethoxy-1, 4-naphthoquinone (DMNQ) in inducing the apoptosis of leukemia cell line U937 and its possible mechanism.</p><p><b>METHODS</b>Flow cytometry and electron microscopy were applied to detect apoptosis of U937 cells after treatment with various combinations of As(2)O(3), DMNQ and ascorbic acid for 24 hours.</p><p><b>RESULTS</b>As(2)O(3) and DMNQ induced-apoptosis of U937 cells was enhanced (35.24%-->61.20%) upon cotreatment with ascorbic acid. Catalase could reverse this effect of DMNQ. Ascorbic acid had no effect on DMNQ-induced apoptosis of U937 cells.</p><p><b>CONCLUSION</b>Ascorbic acid enhanced the apoptosis of U937 cells via reactive oxygen species-dependent pathway in the presence of As(2)O(3).</p>
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Humans , Apoptosis , Arsenicals , Pharmacology , Ascorbic Acid , Pharmacology , Drug Synergism , Flow Cytometry , Naphthoquinones , Pharmacology , Oxides , Pharmacology , U937 CellsABSTRACT
<p><b>OBJECTIVE</b>To explore the association of inherent cellular reactive oxygen species (ROS) levels with susceptibility of the tumor cells to apoptosis induction by arsenic trioxide (As(2)O(3)).</p><p><b>METHODS</b>Low concentration (2 micromol/L) of As(2)O(3) was administered to two cultured leukemic cell lines, NB4 and U937, and two esophageal carcinoma cell lines, EC1.71 (also named EC/CUHK1) and EC1867, to confirm the difference in apoptosis susceptibility of NB4 versus U937 and of EC1.71 versus EC1867. Dihydrogenrhodamine 123 (DHR123), used as a ROS capture agent, was incubated with cells in the absence of As(2)O(3). Fluorescence intensity of rhodamine 123, the product of cellular oxidation of DHR123, was detected by flow cytometry and ROS was measured.</p><p><b>RESULTS</b>Low concentration of As(2)O(3) induced apoptosis was more likely to occur in NB4 and EC1.71 cells than in U937 and EC1867 cells, or NB4 was more sensitive than U937, and EC1.71 more sensitive than EC1867 to As(2)O(3). The inherent cellular ROS level is higher in NB4 than in U937, and also higher in EC1.71 than in EC1867.</p><p><b>CONCLUSIONS</b>The difference in cellular ROS level is positively associated with cellular susceptibility to apoptosis induction by As(2)O(3). The inherent ROS level might be important in defining apoptotic susceptibility to As(2)O(3).</p>
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Apoptosis , Genetics , Arsenicals , Pharmacology , DNA, Neoplasm , Genetics , Flow Cytometry , Fluorescent Dyes , Oxides , Pharmacology , Reactive Oxygen Species , Metabolism , Rhodamine 123 , Tumor Cells, Cultured , MetabolismABSTRACT
Purpose:To explore the association of the inherent cellular level of reactive oxygen species (ROS) with the susceptibility of the tumor cells to apoptosis induced by arsenic trioxide (As 2O 3). Methods:Low concentration(2 ?mol/L) of As 2O 3 was administrated to a pair of the leukemic cell lines, NB4 versus U937, and a pair of the esophageal carcinoma cell line, EC/CUHK1 versus EC1867, to confirm the difference in their susceptibility to apoptosis induced by As 2O 3. Dihydrogenrhodamine123 (DHR123), used as a ROS capture, was incubated with the cells in the absence of As 2O 3 administration. The fluorescent intensity of rhodamine123, which was the product of cellular oxidation of DHR123, was detected by flow cytometry, and ROS was thus measured. Results:For apoptosis induction by 2 ?mol/L of As 2O 3 ,NB4 was more sensitive than U937,and EC/CUHK1 more sensitive than EC1867. The inherent cellular ROS level was higher in NB4 than in U937, and also higher in EC/CUHK1 than in EC1867. Conclusions:The difference in cellular ROS level is associated with the cellular susceptibility to apoptosis induction by As 2O 3.
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Objective To further investigate the balance (relative presence) of Th1 and Th2 subsets at the sites of rheumatoid inflammation,and to understand how about the expression of IL-18 in patients with rheumatoid arthritis (RA),and what the relationship exists between expression of IL-18 and rate of Th1/Th2 in RA,and between IL-18 level and activity of the disease.Method Expression of IFN-γ,IL-4,and IL-18 mRNA in peripheral blood mononuclear cells (PBMC) from 16 patients with RA and 15 healthy subjects was detected by semi-quantitative RT-PCR.Results ① PBMC from patients with RA was found to contain greater levels of IL-18 mRNA than that from healthy subjects (P＜0.01).② IL-18 mRNA levels were correlated with IFN-γ mRNA (relative coefficients:r＝0.836,P＜0.05).③ IL-18 mRNA levels were associated with the disease activity as assessed by levels of erythrocyte sedimentation rate (r＝0.753,P＜0.05).Conclusion IL-18 is among a matrix of inflammatory cytokines produced abundantly in patients with RA and is associated with induction of IFN-γ and activity of the disease.
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Objective To explore the association of the inherent cellular level of reactive oxygen species (ROS) with the sensitivity of the gastric and esophageal carcinoma cells to arsenic trioxide (As 2O 3). Methods The difference of sensitivity to apoptosis induction by low concentration (2 ?mol/L) of As 2O 3 between the gastric carcinoma cell line SGC7901 and MKN45, and between the esophageal carcinoma cell line EC/CUHK1 and EC1867 was firstly demonstrated. SGC7901 was more sensitive than MKN45, and EC/CUHK1 more sensitive than EC1867 to As 2O 3. Then dihydrogenrhodamine 123 (DHR123), as a ROS capture, was incubated with the cells in the absence of As 2O 3. The fluorescent intensity of rhodamine 123, the product of cellular oxidation of DHR 123, was assayed by flow cytometry. The ROS levels was thus detected. Results Inherent cellular ROS level is higher in SGC7901 and EC/CUHK1. They are more sensitive to As 2O 3, than the corresponding cell line MKN45 and EC1867. Conclusions The difference in inherent cellular ROS level of gastric and esophageal carcinoma cells is associated with the cellular sensitivity to apoptosis induced by As2O 3.
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A comparative study of systemic lupus erythematosus in 39 male(MSLE) and 120 female (FSLE) patients was carried out. The results showed that, in MSLE, the mean age at the time of disease onset was similar to FSLE, and the clinical features were nearly the same as those in females, except that the first signs of MSLE were less complicated than those of FSLE, malar rash occurred less commonly in MSLE than in FSLE(P
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To investigate the cellular immunity abnormalities of patients with rheumatoid arthritis(RA), peripheral blood lymphocytes subpopulations, interleukin 2 (IL-2) production and natural killer (NK) cell activity were determined in 9 patients with RA. The results showed that there were a remarkable decrease in NK cell activity and poor response to IL-2 stimlulation. IL-2 production and.the expression of membrane-bound IL-2 receptor were increased in RA patients. The percentage of T4 positive cells and the ratio of T4/T8 were also increased in patients with RA. The results indicate that there are severe cellular immunity abnormalities in patients with RA.