This study was purposed to investigate the effect of high-dose dexamethasone (DXM) on function and Toll like receptor 9 (TLR-9) expression of plasmacytoid dendritic cells (pDC) in peripheral blood of patients with immune thrombocytopenic purpura (ITP). 15 newly diagnosed patients with ITP received high dose DXM at single daily doses of 40 mg for 4 consecutive days. The peripheral blood plasmacytoid dendritic cells from 13 remission patients and 15 normal controls were separated by immunomagnetic beads and then induced by CpG-OND2216. 24 h later, the levels of IFN-α, IL-6 and TNF-α in the supernatant were detected by enzyme linked immunosorbent assay (ELISA). The expression of TLR9 mRNA of pDC was detected by real-time quantitative PCR. The results indicated that the levels of IFN-α, IL-6 and TNF-α produced by pDC in ITP patients were significantly higher than those in normal controls (P < 0.05). After high dose DXM treatment, the levels of IFN-α, IL-6 and TNF-α decreased without significant difference compared with normal controls (P > 0.05). The expression of TLR9 mRNA in pDC of untreated patients was significantly higher than that in control group (P < 0.05), and significantly reduced after treatment without difference from that in control group (P > 0.05). It is concluded that pDC may play an important role in ITP by their TLR9 and secreted cytokines; dexamethasone may down regulate the expression of TLR9, inhibit pDC function, and thus play a therapeutic role.
Adolescent ; Adult ; Case-Control Studies ; Dendritic Cells ; immunology ; metabolism ; Dexamethasone ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood ; drug therapy ; immunology ; RNA, Messenger ; genetics ; Toll-Like Receptor 9 ; genetics ; metabolism ; Young Adult

Objective To analyze the region cluster and risk factors of hypertension in the Chinese adult population and to explore the application of multilevel regression model in the risk factors of hypertension. Methods Multi-stage random sampling technique was used to choose 15 540 individuals aged 35-74 years from 10 regions in China. Two-level logistic regression models were fitted under MLwiN 2.02 software. Results The region cluster of hypertension existed and variance portion coefficient was 3.1%. After adjusting for the age and gender, overall obese people (BMI≥28 kg/m2) were 4.50(95%CI: 4.00-5.06) times, overweight people (BMI=24-27.9 kg/m2) were 2.26 (95%CI: 2.07-2.46) times more likely to be hypertensive as compared with those of normal BMI (18.5-23.9 kg/m2), and those centrally obesive people (Waist circumference≥85 cm in male or 80 cm in female) were 2.62 (95%CI: 2.42-2.83) times more likely to be hypertensive as compared with those of normal WC. The age-and gender-adjusted odds ratios (Ors) of triglyceride (TG), serum total cholesterol (TC), glucose, low-density lipoprotein cholesterol (LDL-C) , high-density lipoprotein cholesterol (HDL-C) and drinking alcohol were 2.10 (95% CI: 1.89-2.33) , 2.08 (95% CI: 1.84-2.35) , 1.85 (95% CI: 1.60-2.14) , 1.58 (95% CI: 1.38-1.81), 1.49(95%CI: 1.32-1.69) and 1.15(95%CI: 1.05-1.27), respectively. Conclusion The prevalence of hypertension was not only affected by individual risk factors, such as obesity, drinking alcohol, abnormal glucose and serum lipids profile, but also affected by the geographic environment where people resided in. Population-and risk factors targeted strategies, proved a promising way to reduce individual risk of hypertension in the primary prevention of hypertension.

OBJECTIVETo investigate the proportion of Th17 cells in the peripheral blood of the patients with acute myeloid leukemia (AML) and evaluate the potential association of Th17 cells with AML.

METHODSThe cytokines IL-17 and TGF-β1 in the peripheral blood of AML patients before therapy (group 1), AML patients in complete remission (AML-CR, group 2) and healthy donors (group 3) were measured by enzyme-linked immunosorbent assay (ELISA). The proportion of Th17 cells of each group was evaluated by flow cytometry. The level of IL-17 mRNA of each group was examined by reverse transcription-PCR (RT-PCR).

RESULTSThe percentage of Th17 cells and the level of IL-17, IL-17 mRNA in group 1 \[(10.502 ± 1.071) ng/L, (0.935 ± 0.140)% and 0.262 ± 0.510\] and group 2 \[(11.345 ± 0.987) ng/L, (1.091 ± 0.159)% and 0.307 ± 0.031\] was significantly lower than that in group 3 \[(16.852 ± 1.198) ng/L, (2.586 ± 0.235)% and 0.501 ± 0.060\]. The percentage of Th17 cells and the level of IL-17, IL-17 mRNA in group 1 was lower than that in the group 2. But the level of TGF-β1 in the group 1 (29.963 ± 1.588) ng/L and the group 2 (25.163 ± 1.848) ng/L was significantly higher than that in group 3 (13.366 ± 1.565) ng/L. However, the level of TGF-β1 in the group 3 was higher than that of the group 2.

CONCLUSIONTh17 cells might be negatively correlated with the AML development. The overexpression of TGF-β1 in AML patients might suppress the differentiation of Th17 cells.

Enzyme-Linked Immunosorbent Assay ; Humans ; Interleukin-17 ; Leukemia, Myeloid, Acute ; Prevalence ; Th17 Cells ; Transforming Growth Factor beta1

OBJECTIVETo explore the changes of surface antigen and function of rituximab on dendritic cells derived from patients with Primary immune thrombocytopenia (ITP) to further understand the effective mechanism of immunotherapy.

METHODSThe peripheral blood mononuclear cells (PBMCs) were isolated from remission patients with ITP before and after low-dose rituximab infusion, and the PMNCs were stimulated for 5 days by rhGM-CSF and rhlL-4 in 5% CO2 air at 37°C incubator. Then all of DCs were cultured with TNF-α for 48 hours. The morphology of DCs was monitored under inverted microscope daily, and the surface antigens of the DCs were analysed by flow cytometry, meanwhile the levels of IL-12p70 and TGF-β1 in supernatants were detected by ELISA, mix lymphocyte reaction was performed by MTT assay.

RESULTS(1) Rituximab-treated-DCs showed no obvious tree-like protruding compared with untreated-DCs. The former cells were small and most of nucleus were centric. (2) The expressions of HLA-DR, CD80, CD83 and CD86 on rituximab-treated-DCs \[56.37 ± 3.95)%, (36.41 ± 2.82)%, (30.45 ± 4.61)% and (41.98 ± 4.17)%, respectively\] were significantly lower than those untreated-DCs \[(73.71 ± 7.61)%, (55.14 ± 7.30)%, (80.91 ± 7.09)% and (59.03 ± 3.43)%, respectively\](all P < 0.05), the concentration of IL-12p70 was significantly lower, \[(66.87 ± 4.29)% vs (50.17 ± 14.52)%\], while that of TGF-β1 \[(9.70 ± 0.31)%\] higher than the untreated-DCs \[(2.70 ± 0.36)%\] (P < 0.05). (3) The abilities to activate T cells proliferation of rituximab-treated-DCs reduced compared with untreated-DCs.

CONCLUSIONThe surface antigen of ITP-DCs and the concentration of IL-12p70 reduced after the low-dose rituximab infusion. The abilities to activate T cells proliferation reduced while the concentration of TGF-β1 increased. Rituximab may achieve its therapeutic effect on ITP by downregulating the immunoreactivity of DCs.

Antibodies, Monoclonal, Murine-Derived ; administration & dosage ; therapeutic use ; Cell Proliferation ; Cells, Cultured ; Dendritic Cells ; cytology ; metabolism ; secretion ; Female ; Humans ; Interleukin-12 ; metabolism ; Lymphocyte Activation ; Male ; Rituximab ; T-Lymphocytes ; immunology ; Thrombocytopenia ; drug therapy ; immunology ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo explore the awareness on sever acute respiratory syndrome (SARS) and public health emergencies among general publics.

METHODSA cluster sampling method was implemented in Harbin and Jiagedaqi district of Daxinanling of Heilongjiang province. Research subjects were divided into three groups as city, township and rural areas and were given questionnaires to fill in. Data was analyzed with Epi-data and SPSS.

RESULTS2003 available questionnaires were collected. The general publics well understood the knowledge on public health emergencies and the SARS with the whole recognition rate more than 60 percent. During the epidemics, people in city, town and countryside were calm (71.7%). The rates of attitude towards the government were significantly different among the subjects living with the city, town or rural areas. The city group expressed the highest favor to the government and media, 71.8% of them gave the credit on the control of SARS to the effective method taken by the government and 65.0% of them showed that they had enough confidence on the governmental ability of dealing with crises while the countryside group trusted the hospitals and relative specialists the most.

CONCLUSIONIt is essential for the government to interact and communicate with the publics through media, medical and related institutions when confronting with the public health emergencies. Publicity on health knowledge and coping system on emergency should play key roles in the development of an effective public health system while the government should lead the battle.

China ; Data Collection ; Emergencies ; Health Knowledge, Attitudes, Practice ; Humans ; Public Health ; Rural Population ; Severe Acute Respiratory Syndrome ; Urban Population

OBJECTIVEA lot of studies have suggested that a certain amount of T cells may be involved among cytokine-induced killer (CIK) cells. The aim of the present study was to prove whether an antigen-specific killing effect on tumor cells is involved during the CIKs-induced killing process.

METHODSBone marrow mononuclear cells (BMMNCs) derived from healthy subjects were separately cultured to generate dendritic cells (DC) and CIKs. A human mammary cancer cell line MCF-7/ADR, expressing P-gp antigen, was frozen-thawed and the lysate including P-gp antigen was obtained. The DC pulsed with or without tumor antigen lysate was co-cultured with CIK (pulsed-DC + CIK and DC + CIK), and CIK cultured alone was used as control. The cell phenotype of DC and CIK was analyzed by flow cytometry. The secretion of IL-12 and IFN-gamma was assayed by ELSA. The antitumor effect of the three CIK groups targeted at MCF-7/ADR cells expressing P-gp antigen and MCF-7 cells was detected by MTT.

RESULTSPulsed-DC + CIK group and DC + CIK group showed a higher expression level of DC mature phenotypes than those before co-culture with CIK, with a significant difference (P = 0.003, P = 0.001, respectively). The phenotypes (CD3, CD8, CD56) of CIK in pulsed-DC + CIK group and DC + CIK group was higher than those in CIK group (P = 0.003, P = 0.011, respectively). Among the three CIK groups, pulsed-DC + CIK group had the highest phenotypes on CD3+ CD56 (pulsed-DC + CIK vs. DC + CIK, P = 0.001; pulsed-DC + CIK vs. CIK, P < 0.001) and CD3 CD8 (P = 0.002, P = 0.002, respectively). Among the three groups, the pulsed-DC + CIK group showed the lowest CD45RA phenotype (pulsed-DC + CIK vs. DC + CIK, P < 0.001; pulsed-DC + CIK vs. CIK, P = 0.004). Among the three groups the secretion of IL-12 and IFN-gamma had the highest level in pulsed-DC + CIK group, with a value of 254 +/- 14.5 pg/ml and 3100 +/- 286 pg/ml, respectively. The antitumor killing effect on MCF-7/ADR cells had a significant difference between any two groups (pulsed-DC + CIK VS. DC + CIK, P = 0.039; pulsed-DC + CIK VS. CIK, P = 0.002; DC + CIK vs. CIK, P = 0.049). The highest was in pulsed-DC + CIK group and the lowest was in CIK group. The CIK group showed a significantly lower antitumor effect on MCF-7 cells than the other two groups (pulsed-DC + CIK vs. CIK, P = 0.007; DC + CIK vs. CIK, P = 0.048), but no significant difference between the pulsed-DC + CIK and DC + CIK groups.

CONCLUSIONIn the present study, DC and CIK cells have been successfully obtained and cultured from bone marrow mononuclear cells. After their co-culture, not only both their specific phenotypes were increased, but also the associated cytokines were secreted. An improved antitumor killing effect and some possible specific immunocytotoxicity were observed. Our findings provided a basis for experimental and clinical research on bio-immunotherapy targeted at multi-drug resistant tumor cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; CD3 Complex ; metabolism ; CD56 Antigen ; metabolism ; CD8 Antigens ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Coculture Techniques ; Cytokine-Induced Killer Cells ; cytology ; immunology ; metabolism ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; metabolism ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism

The tandem mass spectrum of apigenin-6,8-C-di-glucoside( 1) and apigenin-6-C-glucose-8-C-rhamnoside( 2) were obtained by high resolution electrospray ionization mass spectrometry( HR-ESI-MS/MS) in both positive and negative ion modes. The elemental composition of each ion was determined according to its accurate mass-to-charge,hence,the fragmentation pathways of each compound were proposed in both negative and positive ion modes. Comprehensive analysis of each ion and its proposed fragmentation pathways of the two compounds was initially conducted in both negative and positive ion mode HR-ESI-MS/MS to explore the diagnostic ions for flavone-6,8-C-di-glycosides and the characteristic ions for each compound and their cleavage rules. The results showed that a family of fragmentation ions with m/z 353,325,311,297 in ESI(-)-MS and m/z 355,325,307,295 in ESI( +)-MS could be the diagnostic ions of flavone-6,8-C-di-glycoside,and characteristic neutral loss could be assigned to glycosyl substitution,for example,neutral losses of C_4H_8O_4( 120),C_3H_6O_3( 90),C_2H_4O_2( 60) for glucoside substitution while neutral losses of C_4H_8O_3(104),C_3H_6O_2( 74),C_2H_4O( 44) for rhamnoside substitution. Furthermore,only one H_2O loss from mother ion( [M-H]-) was observed for 1 & 2 in ESI(-)-MS while five to six H2 O loss from mother ion( [M+H]+) was observed for 1 & 2 in ESI( +)-MS to produce a family of ions by subsequent loss of H_2O,which could be applied for glucosyl difference. The flavone-6,8-C-di-glycosides in both ESI( +)-MS and ESI(-)-MS showed the cleavage similarity at sugar substitutions. However,there were much more differences by the fragmentation pathways and neutral losses between ESI( +)-MS and ESI(-)-MS as following,hyperconjugation ions by subsequent loss of H_2O from precursor ions of flavone-6,8-C-di-glycosides in ESI( +)-MS were not observed in ESI(-)-MS; the subsequent neutral loss of CH_2O in ESI( +)-MS were rarely observed in ESI(-)-MS; the loss of CO only happen at C-ring of flavone ESI( +)-MS other than glycosyl position in ESI(-)-MS; the C4-chain neutral loss of flavone-6,8-C-di-glycosides happened at 8-C-glycosyl position other than at 6-C-glycosyl position. The above cleavage rules and diagnostic ions of ESI( +)-MS were successfully applied for the structure identification of 4 flavone-6 C,8 C-diglycosides from the stem extract of Dendrobium officinale as vicenin Ⅱ,vicenin Ⅰ,isoschaftoside,schaftoside as well as one flavone-O-glysoside named rutin,which were supported by ESI(-)-MS data as well.
Flavones/chemistry* ; Glycosides/chemistry* ; Ions ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

Objective:To investigate the regulatory effect of Sijunzi Tang(SJZT) and its single herbs(Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma Praeparata cum Melle, Atractylodis Macrocephalae Rhizoma and Poria) on intestinal flora in spleen-deficient rats. Method:Normal rats were randomly divided into the blank group, model group, Zhengchangsheng granules group, SJZT group and each single herb group, rats were orally administered Sennae Folium decoction to induce diarrhea for ten consecutive days to establish a spleen-deficient model(distilled water for the blank group), then treated with the corresponding drugs for seven consecutive days(distilled water for the blank group and the model group). Fresh feces were collected on pre-modeling(0^th day), post-modeling(11^th day), and post-treatment(18^th day). Short-chain fatty acids(SCFAs) in feces were acidified by sulphuric acid and extracted by diethyl ether, then determined by gas chromatography. The structural change(diversity and similarity) of intestinal flora in feces was analyzed by 16S rDNA-polymerase chain reaction(PCR)-denaturing gel gradient electrophoresis(DGGE) technique. Result:Compared with blank group, the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of all administered groups were significantly decreased on the 11^th day(P<0.05, P<0.01), this indicated that the spleen-deficient model was successfully constructed. On the 18^th day, compared with model group, the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of Atractylodis Macrocephalae Rhizoma group were significantly increased(P<0.01), these indexes in feces of SJZT group, Ginseng Radix et Rhizoma group and Poria group were significantly increased(P<0.05, P<0.01), while Glycyrrhizae Radix et Rhizoma Praeparata cum Melle group only increased diversity index of intestinal flora(P<0.05). Conclusion:Ginseng Radix et Rhizoma, Atractylodis Macrocephalae Rhizoma and Poria are the single herbs responsible for the regulatory effect of SJZT on intestinal flora in spleen-deficient rats, and Atractylodis Macrocephalae Rhizoma may play a major role.

In order to explore the compatible principles of Xiebai decoction family, formulae from ancient and modern Xiebai decoction family were collected and sorted in this study. The compatible characteristics, core herbs, as well as the relativity of herbs nature in Xiebai decoction family were analyzed based on scale free network and other data-mining methods such as association rules, clustering analysis and correspondence analysis. The scale free network results showed that in Xiebai decoction family, Mori Cortex-Lycii Cortex-Glycyrrhizae Radix et Rhizoma was used as the core compatible group and formed the complicated compatible network with other additional herbs; association rules results showed that the core herbs in such formulae included Mori Cortex, Lycii Cortex, Glycyrrhizae Radix et Rhizoma, scutellaria root, Platycodon root, Anemarrhena, and almond, which formed corresponding herbal pairs and compatibility; clustering analysis showed that Mori Cortex was the core herb in Xiebai decoction family, and Mori Cortex-Lycii Cortex-Glycyrrhizae Radix et Rhizoma was its main combination unit, which was always compatible with herbs of clearing heat, reducing phlegm, supplementing Qi and nourishing Yin to form the series prescriptions. The results indicated that the core compatibility features of Xiebai decoction family were clearing heat in lung and relieving cough and asthma, providing a basis for the clinical application of Xiebai decoction family.

OBJECTIVETo retrospectively analysis the treatment characteristics of the systemic situation in patients with crush syndrome after Wenchuan earthquake happened in May 12th, 2008.

METHODSForty-nine patients with crush syndrome and subsequent acute renal failure (ARF) due to the earthquake were treated in West China Hospital. All of patients had been rescued from buildings that collapsed in Wenchuan earthquake. The major associated injuries were in the low extremities and upper extremities. 49 patients developed ARF with increased concentrations of serum creatinine (mean 64 022 U/L) had underwent haemodialysis. Hyperkalaemia was seen in 9 patients and four of them underwent haemodialysis. 49 patients were administered hemodialysis.

RESULTSNo patient died. All patients who suffered from the ARF were weaned from hemodialysis after admitted 7 to 35 days. Forty-five extremities underwent amputations and 52 extremities had fasciotomy.

CONCLUSIONSCrush syndrome requires urgent recognition and prompt surgical treatment with simultaneous measures to control hyperkalemia and ARF. The authors believe that immediate intensive care therapy and multi-subjective coordination would have improved the survival rate.

Acute Kidney Injury ; etiology ; surgery ; therapy ; Adolescent ; Adult ; Aged ; Amputation ; Child ; Crush Syndrome ; etiology ; surgery ; therapy ; Decompression, Surgical ; Earthquakes ; Female ; Humans ; Male ; Middle Aged ; Renal Replacement Therapy ; Retrospective Studies ; Treatment Outcome ; Wounds and Injuries ; complications