Objective To detect and determine the specificity of platelet-reactive antibodies in patients who were refractory to platelet transfusions.Methods Serum samples from 48 patients who were refractory to platelet transfusions were screened with MACE for platelet-reactive antibodies.Specificity of platelet alloantibodies was determined with PAK12 and MAIPA.Results Platelet-reactive antibodies were detected in the serum of 50% PTR patients(24/48).The incidence of HLA antibodies was 39.6%(19/48),accounting for 79.2% of serum with platelete alloantibodies.The HPA alloantibodies were found in 29.2%(14/48)serum,of which,64.3%(9/14)occurred together with anti-HLA.The following platelet-specific antibodies were identified:anti-HPA-3a(n=2),anti-HPA-5b(n=1),anti-HPA-5a(n=1),anti-HPA-2b(n=1),anti-HPA-4b(n=1).Of the 14 serum with HPA antibodies,78.6%(11/14)contained panreactive anibodies against platelet glycoprotein(GP)Ⅱb/Ⅲa,GPⅠa/Ⅱa,and/or GPⅠb/Ⅸ.Platelet-reactive antibodies were detected more in female(16/29)than in male(8/19)with a frequency of 55.2%,42.1%,respectively,but there was no statistical significant difference.Conclusion The platelet-specific antibody in PTR patients are not as rare as previous thought although alloantibodies are predominantly anti-HLA.Antibody specificities in Chinese PTR patients are different from those observed in Caucasians,in whom anti-HPA-5b and-1b are the most prevalent specificity.The most prevalent platelet-specific antibodies are anti-HPA-3 and anti-HPA-5 while anti-HPA-4b and anti-HPA-2b are also detected.

0.5).The estimated daily intakes of 10 nutrients were analyzed, and showed that the protein, calcium and saturated fatty acids increase the risk of stomach cancer (P

AIM:To explore an ideal method to induce the differen-tiation of human umbilical cord mesenchy-mal stem cells (hUCMSCs) into neuron-like cells and to provide some evidence for the transplantation of hUCMSCs for spi-nal cord injury .METHODS:The hUCMSCs were isolated from human umbilical cord digested with collagenase Ⅱ.The hUCMSCs was verified by flow cytometry analysis .The passage 5 cells were randomly divided into 4 groups.The differentiation of hUCMSCs was induced by bFGF in group A , bFGF and BDNF in group B, or BHA, bFGF and BDNF in group C, while the cells in group D served as a control group cultured with DMEM-F12 and 10%FBS.Two weeks later , the expression of nestin , neurofilament protein H ( NEFH) and glial fibrillary acidic protein ( GFAP) was detected by real-time PCR and immunocytochemistry .The morphological changes of cells were observed under an atomic force microscope . RESULTS:Mesenchymal stem cells were isolated and cultured from human umbilical cord by enzyme digestion .hUCMSCs expressed CD29, CD44 and CD105, but no CD34, CD45 or HLA-DR.After cultured with inducing medium for 2 weeks, the cells were successfully induced into neuron-like cells.The appearance of the cells had great change .The induced hUC-MSCs developed round cell bodies with multiple neurite-like extensions observed under an atomic force microscope .The re-sult of real-time PCR showed that nestin was positive in A , B and C groups , and NEFH was positive in A and B groups , but GFAP was negative in 4 groups.The difference of nestin and NEFH expression among the induced groups was signifi -cant (P<0.05).CONCLUSION:Mesenchymal stem cells were isolated and cultured from human umbilical cord by en-zyme digestion in vitro, and all the hUCMACs presented stable biological properties .Moreover, hUCMSCs were induced to differentiate into neuron-like cells in vitro via bFGF combined with BDNF .

Objective Helicobacter pylori（ ）， To investigate the infection status of HP and analyze the correlation between HP Methods infection and serum bilirubin in railway drivers. A total of 2 731 railway drivers in Zhengzhou locomotive depot were - selected as study subjects using judgment sampling method. Carbon 13 urea breath test was used to evaluate the HP infection ， status. The metabolic indexes of HP positive group and HP negative group were compared and the relationship between HP Results （ ） ， infection and serum bilirubin was analyzed. The HP infection rate was 42.3% 1 156/2 731 . The older the age the ， （ ）， （ P ） longer the work years and the higher the body mass index BMI the higher the HP infection rate all <0.01 . The infection （P ） rate of HP in married people was higher than that in unmarried people <0.01 . The HP infection rate of smokers was higher - （P ） - ， than that of non smokers <0.01 . Compared with the HP negative group fasting blood glucose and serum levels of total ， （ - ）， （ ） - cholesterol low density lipoprotein cholesterol LDL C triglyceride and homocysteine Hcy were increased in the HP （ P ） （ - ）， ， positive group all <0.05 . The serum levels of high density lipoprotein cholesterol HDL C total bilirubin direct bilirubin （ ） - （ P ） DBIL and indirect bilirubin were lower than those in HP negative group all <0.05 . Logistic regression analysis showed that （ P ） HP infection was associated with low serum total bilirubin and low DBIL all <0.01 after adjusting for the confounding effects ， ， ， ， ， ， ， - ， - ， of age work years marital status smoking history fasting blood glucose total cholesterol triacylglycerol LDL C HDL C Conclusion ， ， ， and Hcy. The age work length BMI smoking and marital status are the influencing factors of HP infection in railway drivers. HP infection is associated with low levels of total bilirubin and DBIL.

In this paper, the chloroplast psbK-psbI intergenic spacers of 18 species of Dendrobium and their adulterants were amplified and sequenced, and then the sequence characteristics were analyzed. The sequence lengths of chloroplast psbK-psbI regions of Dendrobium ranged from 474 to 513 bp and the GC contents were 25.4%-27.6%. The variable sites were 71 while the informative sites were 46. The inter-specific genetic distances calculated by Kimura 2-parameter (K2P) of Dendrobium were 0.006 1-0.058 1, with an average of 0.028 4. The K2P genetic distances between Dendrobium species and Bulbophyllum odoratissimum were 0.093 2-0.120 4. The NJ tree showed that the Dendrobium species can be easily differentiated from each other and 6 samples of the inspected Dendrobium species were identified successfully through sequencing the psbK-psbI intergenic spacer. Therefore, the chloroplast psbK-psbI intergenic spacer can be used as a candidate marker to identify Dendrobium species and its adulterants.
Chloroplasts ; DNA, Chloroplast ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Plants, Medicinal ; classification ; genetics

Objective To investigate the effects of macrophage inflammatory protein-1α (MIP-1α) combined with molecule 4-1BB L on the tumorigenicity of hepatocellular carcinoma cells in vivo. Methods Mouse MIP-1α (mMIP-1α) expressed Hepa 1-6 cells were transfected with m4-1BBL recombinant retrovirus, the anti-histidinol cells clones were selected and amplified. The expression of m4-1BB L was confirmed by flow cytometry. The growth curve of Hepa 1-6 cells transfected with mMIP-1α and m4-1BBL alone or together was drawn and compared. C57B/L Mice were randomly divided into 7 groups, 9 mice in each group, injected with mMIP-1α+m4-1BB L Hepa 1-6 cells, m4-1BB L Hepa 1-6 cells, mMIP-1α Hepa 1-6 cells, Hepa 1-6 cells, pLXSHD Hepa 1-6 cells or PBS respectively. The tumorigenicity of hepatocellular carcinoma cells and the mice survival rate were compared between each groups. Results Hepa 1-6 mMIP-1α+m4-1BB L cells which expressed both mMIP-1α and m4-1BB L were successfully established. The expression of mMIP-1α and m4-1BB L alone or together did not affect the growth curve of Hepa 1-6 cells. Observed for 5 weeks, no tumor developed in Hepa 1-6 mMIP-1α+m4-1BB L injected mice. The tumorigenicity of Hepa 1-6 mMIP-1α+m4-1BB L was lower than that of Hepa 1-6 mMIP-1α or Hepa 1-6 m4-1BB L in vivo. The survival rate of Hepa 1-6 mMIP-1α+m4-1BBL injected mice(9/9) was higher than that of Hepa 1-6 m4-1BB L injected mice (6/9)or Hepa 1-6 mMIP-1α injected mice (1/9). Conclusion Chemokine MIP-1α combined with costimulatory 4-1BB L lowered the tumorigenicity of hepatocellular carcinoma cells in vivo, and prolonged the mice survival period.

OBJECTIVE Recent studies have demonstrated that the Nlrp3 inflammasome serve as a central role in the pathogenesis of cardiovascular diseases and endothelial dysfunction occurs in association with several cardiovascular risk factors. Given the demonstrated anti-inflammatory effects of aspirin, the present study was designed to test whether aspirin diminish NLRP3 inflammasome activation and prevent endothelium injury and associated coronary artery damage during LPS. METHODS Mouse carotid arterial endothelial cells (CAECs) were cultured and treated with 0.1-3 mmol·L-1 of aspirin in response to LPS (2 μg·mL-1) stimuli. After 24 h, the Nlrp3 inflammasome complexes consist of varied proteins were analyzed by WB. NO and T-AOC in the supernatant was detected by ELISA. Intracellular reactive oxygen species (ROS) generation for 24 h was observed by DCF fluorescence. The mice were treated with aspirin (12.5 mg·kg-1 per day, 62.5 mg·kg-1 per day, 125 mg·kg-1 per day) and dexametha?sone (0.0182 mg · kg- 1 per day) for 7 d. The level of IL- 1β,IL- 18 protein was detected by ELISA. RESULTS Immunofluorescence results showed the colocalization of Nlrp3 with ASC or caspase 1 decrease in a concentration- dependent manner. Meanwhile, the expression of Nlrp3 and caspase 1 protein was decreased with the concentration of aspirin, but no changes the expression of ASC protein. Nlrp3 protein levels in CAECs were 0.33- 0.8- fold and cle- caspase 1 protein levels in CAECs were 0.48-1-fold compared to those in LPS stimulation when treated with 0.1-3 mmol·L-1 aspirin for 24 h (P<0.01). Aspirin significantly antagonized the effect of LPS on NO (1.22-1.91-fold that of LPS stimulation, P<0.01) and T-AOC expression (1.02-1.90-fold that of LPS stimulation, P<0.01). As the different concentration of aspirin treated, the generation of ROS was 0.51-1.10-fold that of LPS stimulation (P<0.01). In vivo data shown the level of IL-1β, IL-18 protein from serum are in concordance with the level of Nlrp3 inflammasome activation. CONCLUSION We conclude that aspirin has anti- inflammatory properties, protecting CAECs from LPS-induced injury by inhibition of NLRP3 inflammasome activation through ROS pathway.

Objective: To investigate the effects of andrographolide on extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and tumor necrosis factor-α (TNF-α) expression in lipopolysaccharide (LPS)-activated macrophages. Methods: LPS-activated mouse peritoneal macrophages were cultured in media with different concentrations of andrographolide. Cytotoxicity of andrographolide was detected by cell counting kit-8. The macrophages were lysed, and then expressions of phosphorylated ERK1/2, JNK and p38 and nuclear factor-κB inhibitor (IκBα) protein were detected by Western blotting and TNF-α mRNA expression was detected by reverse transcription-polymerase chain reaction. Supernatants of the macrophages were used to detect content of TNF-α protein by enzyme-linked immunosorbent assay. Results: Andrographolide at 1-100 μg/mL showed no cytotoxicity on LPS-activated mouse peritoneal macrophages. Andrographolide inhibited ERK1/2 phosphorylation in LPS-activated murine peritoneal macrophages, which was concentration-dependent (P<0.01). Andrographolide at 1-25 μg/mL had no effects on phosphorylation levels of JNK and p38 and IκBα degradation in LPS-stimulated mouse peritoneal macrophages. In activated macrophages, TNF-α expression was inhibited by 12 μg/mL andrographolide and 20 μmol/L PD98059 (inhibitor of ERK1/2 signaling pathway) at both mRNA expression and protein secretion levels. Conclusion: In LPS-activated macrophages, andrographolide may inhibit the expression of TNF-α by inhibiting ERK1/2 signaling pathway.

OBJECTIVETo investigate the clinicopathological characteristics of gastrointestinal tract involvement of anaplastic large cell lymphoma (ALCL).

METHODSThe clinicopathological features of four patients with ALCL that involved gastrointestinal tract were retrospectively analyzed using immunohistochemical study, T-cell receptor gene rearrangement analysis, and evaluation for Epstein Barr virus infection status.

RESULTSMost tumor cells in all these four cases are large and highly pleomorphic, and all four cases were classified as the common pattern ALCL. Tumor cells in all four tumors expressed CD30, and expressed at least one cytotoxic maker. Two patients were confirmed to be with anaplastic lymphoma kinase (ALK)-positive ALCL, and four patients were negative during in situ hybridization for Epstein-Barr virus-encoded RNA but showed clonal T-cell receptor gene rearrangement.

CONCLUSIONGastrointestinal tract involvement of ALCL has the unique clinicopathological features.

Adult ; Biomarkers, Tumor ; metabolism ; Epstein-Barr Virus Infections ; Female ; Gastrointestinal Neoplasms ; diagnosis ; pathology ; Gene Rearrangement, T-Lymphocyte ; Humans ; Ki-1 Antigen ; metabolism ; Lymphoma, Large-Cell, Anaplastic ; diagnosis ; pathology ; Male ; Retrospective Studies

OBJECTIVETo summarize the clinical features, diagnostic and therapeutic experiences of colorectal Non-Hodgkin's lymphoma (NHL).

METHODSClinical data of 32 patients with colorectal NHL admitted to our hospital from January 1988 to December 2006 was retrospectively analyzed.

RESULTSThis study included 22 B-cell NHL and 10 T-cell NHL cases. In the B-cell NHL group, the male: female ratio was 14:8 and the median age was 60.5 years. In the T-cell NHL group, the male: female ratio was 5:5 and the median age was 31.0 years. The ileocecal region was most frequently involved in both groups, which accounted for 77.3% and 60.0% of the B and T group respectively. The common clinical manifestations included abdominal pain, weight loss, and abdominal mass. Of the 14 cases of B-cell NHL with definite subtype classifications, 64.3% were of the Diffuse Large B-cell Lymphoma (DLBCL) type. Among the 22 B-cell NHL, 40.9% were with localized diseases (stage I-II1), while all 10 patients in T-cell NHL group were in stage IV with 3 patients complicated with massive GI bleeding and 4 with perforation. All patients of B-cell type received chemotherapy utilizing mainly CHOP after surgical resection. After a median follow-up of 55 months, the disease-free survival was rate 88.2%. Among the T-cell NHL group, 8 out of 10 patients underwent surgery and chemotherapy was given to all those who could tolerate it. Five patients died within 2 months after surgery. It's known that 3 patients were still alive after 23 months.

CONCLUSIONSThe ileocecal region is the most frequently involved site of the colorectal NHL. The histology is usually B-cell type with a majority being DLBCL. Currently R-CHOP chemotherapy after the surgical resection is the principal treatment modality. Patients of B-cell type have a better prognosis while the prognosis of T-cell NHL is poor. Therefore more aggressive diagnostic and therapeutic approaches are recommended for T-cell NHL patients. The prospective of organ preservation treatment for colorectal NHL is still in need of further investigations.

Adult ; Aged ; Colorectal Neoplasms ; diagnosis ; therapy ; Female ; Follow-Up Studies ; Humans ; Lymphoma, Non-Hodgkin ; diagnosis ; therapy ; Male ; Middle Aged ; Retrospective Studies