Objective To evaluate the effect of propofol on interleukin-1β (IL-1β)-induced increase in monolayer permeability of human umbilical vein endothelial cells (HUVECs).Methods Primary HUVECs were cultured and purified by immuno-magnetic separation.The expression of VE-cadherin in endothelial cells was determined by immunofluorescence.The HUVEC monolayer permeability was detected by the Transwell system.The cells were seeded on the upper chamber (2 × 105 cells/well) and cultured for 3 days after confluence.The cells were treated in two ways.The cells were randomly divided into 6 groups (n =36 each) and 5 of the 6 groups treated with 1,2,5,10 and 20 ng/ml IL-1β for 24 h except for control group.The cells were also randomly divided into 5 groups (n =30 each) and 4 of the 5 groups were pretreated with 0,10,50 and 100 μmol/L propofol for 30 min,and then treated with 10 ng/ml IL-1β for 24 h except for control group.The cells were radomly divided into 3 groups (n =18 each) and 2 of the 3 groups were pretreated with 50 μmol/L propofol for 30 min,and then treated with 10 ng/ml IL-1β for 24 h or 30 min.The expression of occludin protien,p38 mitogen activiated protienkinase (p38 MAPK) and phosphorylated p38 MAPK (p-p38 MAPK) was determined by Western blot.Results Compared with control group,5,10 and 20 ng/ml IL-1β significantly increased HUVEC monolayer permeability in a concentration-dependent manner (P ＜ 0.05 or 0.01).10,50 and 100 μmol/L propofol inhibited IL-1 β-induced increase in the permeability of HUVEC monolayer permeability in a concentration-dependent manner (P ＜ 0.01).IL-1β could down-regulate HUVEC occludin protein expression,and activate p38MAPK signaling pathway,and propofol inhibited IL-1β-induced down-regulation of HUVEC occludin protein expression and activation of p38 MAPK signaling pathway (P ＜ 0.01).Conclusion Propofol can alleviate IL-1β-induced increase in the permeability of HUVEC monolayer via inhibiting activation of p38 MAPK signaling pathway.

Objective To analyze the association of Crohn's disease(CD)with vitamin D receptor(VDR) gene polymorphisms. Methods After collecting 326 CD patients and 464 healthy controls,the four single nucleotide polymorphisms of VDR (FokI, BsmI, ApaI and TaqI) were examined by a SNaPshot technique. Results Compared with those in controls,the frequencies of mutant allele(A)and genotype(GA+AA)of BsmI were significantly decreased in CD patients(both P=0.001). The similar conclusions were also drawn for the mutant allele(C)and genotype(TC+CC)of TaqI(both P<0.05). In further stratified analysis,compared with those in controls,the mutant alleles and genotypes of BsmI and TaqI were significantly reduced in stenotic type CD patients (all P<0.0083). The analyses of linkage disequilibrium(LD)and haplotype showed that BsmI,ApaI and TaqI were in a strong LD,and the formed haplotype AAC was significantly lower in CD patients than that in controls (P <0.05). Conclusions VDR(BsmI and TaqI)polymorphisms are significantly related with the reduced susceptibility to CD,especially for patients with stenotic CD. Moreover,the haplotype AAC might engender a reduced risk of CD.

OBJECTIVETo examine the correlation between the mRNA and proteins expressions of gastrin(GAS), and the association of protein expression of GAS with apoptosis index(AI) and apoptosis regulation gene Fas/FasL, caspases in colorectal cancer.

METHODSThe expressions of GAS mRNA in tumor tissues of 79 cases with colorectal cancer were detected by nested RT-PCR. Cell apoptosis was detected by molecular biology in situ apoptosis detecting technic(TUNEL). Protein expressions of GAS, Fas/FasL, and caspases were detected by immunohistochemical staining (SP method).

RESULTSThe positive correlation was found between the mRNA and proteins expressions of GAS(rGAS=0.99, P<0.01). The mRNA and protein expressions of GAS in well and moderately differentiated cancers were significantly lower than those in poorly differentiated cancers (chi(2)(high vs low)=10.47, 10.23, P<0.01, chi(2)(middle vs low)=6.68, 4.95, P<0.05). The mRNA and protein expressions of GAS in papillary and tubular adenocarcinomas were significantly lower than those in mucinous adenocarcinomas, signet-ring cell carcinoma and undifferentiated carcinoma (chi(2)(papillary vs mucinous and signet-ring)=4.80, 6.22, chi(2)(papillary vs undifferentiation)=5.44, 8.43, chi(2)(tubular vs mucinous and signet-ring)=4.40, 4.38, chi(2)(tubular vs undifferentiation)=4.92, 6.43, P<0.05, respectively). The mRNA and protein expressions of GAS in Dukes' stages A, B were significantly lower than those in Dukes stages C, D (chi(2)=4.84, 4.45, P<0.01). The AI in GAS high and moderate expression groups of colorectal cancer were significantly lower than that in low expression group (q(high vs low)=6.71, q(middle vs low)=4.60, P<0.01). The positive expression rate of FasL was significantly different among GAS high, moderate and low expression groups of colorectal cancer (chi(2)=9.35, P<0.01). The positive expression rate of FasL in GAS high and moderate expression groups was higher than that in low expression group (chi(2)high vs low=6.24, chi(2)(middle vs low)=4.74, P<0.05).

CONCLUSIONSGAS plays an important role in the regulation of cell apoptosis in colorectal carcinoma, whose mechanism may be related to the aberrant expression of Fas/FasL. GAS will be one of the indicators of the biological behavior in colorectal carcinoma.

Adult ; Aged ; Caspases ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Fas Ligand Protein ; metabolism ; Female ; Gastrins ; metabolism ; Humans ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Young Adult ; fas Receptor ; metabolism

This study was aimed to analyze the hematologic and molecular biologic characteristics of 14 Wenzhou patients with minor beta-thalassemia, to find out the mutation sites responsible for the disease by detecting sequences of PCR products and to analyze the single nucleotide polymorphism. The peripheral blood of patients was collected intravenously and was anticoagulated with EDTA-K(2); then the templates from blood samples were extracted, the related primers were designed for sequencing the products amplified by PCR; finally mutation sites resulting in beta-thalassemia were found through comparison and analysis of sequences. The results indicated that the C-->T heterozygous mutation occurred at the IVS-2 -654 site in 4 cases; the TTCT deficiency appeared at CD41/42 site in 1 case; in 2 sites existed single nucleotide polymorphisms occurring at the 59th site of exon 1 (T/C, CAT/CAC, His) and IVS-2 nt 665 (T/C). It is concluded that single nucleotide polymorphism of minor beta-thalassemia patients born in Wenzhou had specificity, this study found too kinds of gene mutations which are IVS-2 -654 C-->T heterozygous mutation and CD41/CD42 site-TTCT deficiency.
Base Sequence ; China ; Humans ; Mutation ; Polymorphism, Single Nucleotide ; beta-Globins ; genetics ; beta-Thalassemia ; genetics ; metabolism

To investigate the difference in absorptive of tetrahydropalmatine (THP) and l-tetrahydropalmatine (l-THP) in rat intestine as well as the mechanism of the absorption of THP, in situ single pass perfusion model was used and the concentration of THP in perfusate was determined by HPLC. The absorption rate constant (k(a)) and effective permeability values (P(eff)) of THP had no significant difference (P > 0.05) at concentration of 8, 16 and 32 microg x mL(-1) in perfusion or in four different regions of intestine of rat (duodenum, jejunum, ileum, colon). The absorption of l-THP and THP in jejunum had significant difference (P < 0.05). The k(a) and P(eff) of THP increased obviously when verapamil was co-perfused with THP, while those of l-THP were not influenced by verapamil. The absorption of THP in intestine showed the passive diffusion process, and without a special absorption region. The stereoselective absorption difference may result from stereoselective combination of P-glycoprotein with d-THP.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Animals ; Berberine Alkaloids ; chemistry ; pharmacokinetics ; Colon ; metabolism ; Duodenum ; metabolism ; Female ; Ileum ; metabolism ; Intestinal Absorption ; drug effects ; Jejunum ; metabolism ; Male ; Perfusion ; Permeability ; Rats ; Rats, Sprague-Dawley ; Sex Factors ; Stereoisomerism ; Verapamil ; pharmacology

External snapping hip(ESH) is a vague term used to describe palpable or auditory snapping with hip movements with or without pain. The pathogenesis of ESH is related to the specific anatomical structure and friction factor. The clinical symptom is auditory snapping during activities, physical examination, X-ray, magnetic resonance imaging(MRI), dynamic ultrasound and other imaging techniques can be used to diagnose. Conservative medical management includes rest, avoidance of aggravating activities, and antiinflammatory medications. Treatment Patients with mild symptoms can achieve good results by medication, rest and physiotherapy. Surgical treatment for patients with ineffective conservative treatment was performed. All kinds of open surgery method can achieve good clinical curative effect, arthroscopic surgery is gradually been promoted due to small trauma, less complications. Besides, there are some reports that traditional treatments such as massage, acupuncture and acupotomology have achieved good clinical results, which deserve further study and promotion.

This study was aimed to investigate the curative effect and safety of human umbilical cord mesenchymal stem cells (hUCMSC) to treat acute graft-versus-host disease (aGVHD) of children after hematopoietic stem cell transplantation (HSCT). HUCMSC were isolated and cultured by collagenase digestion and passage culture. The 3rd to the 5th passage of hUCMSC were used for clinical treatment. Five cases of children acute leukemia achieved complete remission after chemotherapy. Two cases received HLA 3/6 loci matched haploidentical bone marrow HSCT. One case received HLA-matched sibling bone marrow and peripheral blood HSCT. One case received unrelated HLA 4/6 loci matched umbilical cord blood HSCT. One case received unrelated HLA 5/6 loci matched umbilical cord blood HSCT. The children received immunosuppressive therapy after III-IV aGVHD occurring. They received 0.5×10(6)/kg hUCMSC infusion when conventional therapy was ineffective. The results showed that 5 cases of children acute leukemia achieved hematopoietic reconstitution and developed the III-IV grade aGVHD. The five cases of children were infused with hUCMSC. The rash subsided, the liver function was normalized and the gastrointestinal symptoms were improved. The infusion-related adverse reaction did not happen. At present, the 5 children are in remission. It is concluded that allogeneic HSCT is an effective therapeutic method for children with acute leukemia. HUCMSC infusion can be safely and effectively used for the treatment of refractory aGVHD.
Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; Female ; Graft vs Host Disease ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Treatment Outcome

OBJECTIVETo assess the prevalence of several tyrosine kinases (TKs) gene mutations including c-Kit, FLT3 and JAK2 V617F in core binding factor related acute myeloid leukemia (CBF-AML), and analyze their impact on clinical characteristics and prognosis.

METHODSMutations of c-Kit, FLT3-ITD and FLT3-TKD were detected by genomic DNA PCR and sequencing, and JAK2 V617F mutation screening by allele-specific PCR in 58 newly diagnosed CBF-AML patients [28 AML with inv(16) and 30 with t(8;21)], and analyze the patients clinical characteristics and prognoses.

RESULTSc-Kit aberrations were detected in 32.8% cases, including 6 cases mutated in exon 8 (mutKIT8) and 13 mutated in exon 17 (mutKIT17). MutKIT8 was more prominent in inv(16) than in t(8;21) patients (21.4% vs 0, P = 0.009). Only 2 cases had FLT3-ITD and 7 (12.1%) FLT3-TKD mutations. The result of JAK2 V617F mutation screenings in these CBF-AML patients was negative. The frequency of receptor tyrosine kinases(RTK) mutations was 46.6% and only one case had two kinds of missense mutations (mutKIT8 & TKD(+)). Median age of onset was higher for mutKIT17 than for wide-type c-Kit (wtKIT) patients (55 vs 31, P = 0.003). c-Kit mutations were significantly associated with decreased overall survival (OS) and continuous complete remission (CCR) rates (P = 0.053, and 0.048 respectively), and so did more for exon17 mutated patients reduced (P = 0.005, and 0.013 respectively). FLT3-TKD mutation showed no effects on prognosis of CBF-AML patients.

CONCLUSIONSRTK mutations are common in patients with CBF-AML. c-Kit mutations frequently and JAK2V617F mutation rarely appear in CBF-AML. c-Kit mutations, especially mutKIT17 confers higher relapse risk and poorer prognosis.

Adolescent ; Adult ; Aged ; Core Binding Factors ; DNA Mutational Analysis ; Female ; Humans ; Janus Kinase 2 ; genetics ; Leukemia, Myeloid, Acute ; diagnosis ; etiology ; genetics ; Male ; Middle Aged ; Mutation ; Prognosis ; Protein-Tyrosine Kinases ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

OBJECTIVETo evaluate the prevalence of TET2 gene mutation in acute myeloid leukemia (AML) patients, and analyze their clinical characteristics and prognosis.

METHODSPolymerase chain reaction (PCR) and direct sequencing were used to sequence exon 3 to 11 of TET2 gene.

RESULTSAmong 96 AML patients, TET2 gene mutation was detected in 13 (13.54%) patients (95%CI 6.70% - 20.38%). The median age was 54 years in mutated group and 41 years in unmutated group (P = 0.010). Mutated and unmutated patients did not significantly differ in gender, white blood cells (WBC) count at diagnosis, platelet count, PB and BM blast percentage and chromosome karyotype, excepting for hemoglobin level 84 (70 - 108) g/L in mutated group versus 70 (55 - 87) g/L in unmutated group (P = 0.032). TET2 gene mutation had no significant correlation with C-KIT, FLT3, JAK2V617F mutations, but did with NPM1 mutation. TET2 mutated patients had lower CR1 rate and 2-year overall survival than unmutated in non-M(3) patients (P < 0.05).

CONCLUSIONSTET2 gene mutation is more prevalent in older AML patients and has a certain correlation with clinical characteristics and outcome. It may be a molecular marker for poor prognosis in AML.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; Exons ; Female ; Humans ; Karyotype ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Proto-Oncogene Proteins ; genetics ; Young Adult

BACKGROUNDPrevious prognosis analyses of colorectal cancer (CRC) patients with stage II and III disease were done as separate categories. The purpose of this study was to analyze prognostic factors associated with survival in a group of patients who underwent radical resection of stages II and III CRC.

METHODSA retrospective review was performed for 141 consecutive stages II and III patients who had undergone radical resection of colorectal adenocarcinoma between May 2003 and November 2003. Univariate and multivariate analyses were performed to assess the effect of record variables on disease free survival and overall survival.

RESULTSThe median follow-up time was 59 months, and the 3- and 5-year survival rates were 76% and 68%, respectively. Four factors were independently associated with a worse disease-free survival: diabetes (hazard ratio (HR) 2.338; 95% confidence interval (CI) 1.011 - 5.407), expression of cyclooxygenase-2 (Cox-2) (HR 0.335; 95%CI 0.126 - 0.888), expression of matrix metalloproteinases 2 (MMP-2) (HR 0.233; 95%CI 0.101 - 0.541), expression of vascular endothelial growth factor (VEGF) (HR 0.295; 95%CI 0.088 - 0.996). Four factors were independently associated with a worse overall survival: lymph nodes metastasis (HR 1.67; 95%CI 1.29 - 2.14), Cox-2 positive (HR 0.056; 95%CI 0.247 - 0.731), MMP-2 positive (HR 0.398; 95%CI 0.190 - 0.836), VEGF (HR 0.364; 95%CI 0.090 - 0.716).

CONCLUSIONSDiabetes, expression of Cox-2, MMP-2 and VEGF were independently associated with a worse disease- free survival. Lymph nodes metastasis, expression of Cox-2, MMP-2 and high level of VEGF predicted a poor overall survival.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Colorectal Neoplasms ; metabolism ; pathology ; Cyclooxygenase 2 ; metabolism ; Disease-Free Survival ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; pathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Middle Aged ; Multivariate Analysis ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult