Objective:To investigate the expression status of Sonic Hedgehog signaling genes and molecules in human hepatocellular carcinomas(HCC),and to explore the relationship between these genes and clinical prognosis.Methods:HCC tissue and adjacent normal tissue from 29 HCC patients were assayed for the expression of hedgehog signaling genes by reverse transcription-polymerase chain reaction chain reaction(RT-PCR) techniques and for the expression of hedgehog signaling molecules by immunohistochemistry.The expressions of Shh,Ptch,Smoh,Gli-1 mRNA were assayed as well as Shh,Ptch proteins in 29 cases of HCC and in 29 liver tissues adjacent to the tumor.Results:Expression of Shh mRNA was detectable in about 51% of HCCs examined.Consistent with this,hedgehog target genes Ptch,Smoh and Gli-1 mRNA were expressed in over 68%,48% and 62% of the tumors,respectively,and the expressions of Shh and Ptch proteins in HCC tumor tissues correlated with those of Shh and Ptch mRNA in tumor tissues(P=0.041 and P=0.035).This suggested that the hedgehog pathway was frequently activated in HCCs.The simultaneous expression of Gli-1 in HCC and liver tissues adjacent to the tumor had significantly relationship with poor prognosis.Conclusion:Hedgehog signaling activation is an important event for development of human HCCs.It also suggests that markers for hedgehog signaling activation may be useful for the determination of prognosis.

OBJECTIVE: To investigate the influence of demographic and clinical characteristics, stress, and coping style on disease self-management in children and adolescents with type 1 diabetes. METHODS: A cross-sectional survey was performed to select 149 children and adolescents with type 1 diabetes (aged 8-20 years). Related data were collected using the questionnaires and scales on general information, diabetes self-management, perceived stress, and coping style. RESULTS: Of the 149 children and adolescents, 37(24.8%) had high stress. Compared with the school-aged children, the adolescents had higher stress level and were more likely to present with negative coping style (P<0.05). The multiple linear regression analysis showed that the children whose mothers had an educational level at or above senior high school, who had a low stress level, and who adopted positive coping measures had a higher level of diabetes self-management (P<0.05). CONCLUSIONS Nearly a quarter of the children and adolescents with type 1 diabetes have a high stress level. When delivering the education on diabetes self-management to children and adolescents, healthcare workers should focus on the families whose mothers have an educational level at or below junior high school. Strategies should aim at reducing stress by encouraging positive coping styles.
Adaptation, Psychological ; Adolescent ; Adult ; Child ; Cross-Sectional Studies ; Diabetes Mellitus, Type 1 ; Female ; Humans ; Self-Management ; Stress, Psychological ; Surveys and Questionnaires ; Young Adult

OBJECTIVE To predict the potential targets of hyperoside (Hyp) on improving ischemia/reperfusion injury by network pharmacology, and explore its possible mechanism combined with related literature. METHODS The action targets of Hyp and ischemia/reperfusion injury were obtained by TCMSP, Swiss Target Prediction, Pharm Mapper, Simi?larity ensemble approach, Online Mendelian Inheritance in Man, DisGENT and database. The common targets of drugs and diseases were screened by Omishare and STRING database respectively, and the protein-protein interaction (PPI) network map was constructed. Then the interaction network between Hyp and disease targets was constructed by Cyto?scape software and topological cross-linking analysis was carried out. Then the interaction network between Hyp and disease targets was constructed and cross-linked analysis was carried out by using Cytoscape software. The gene ontol?ogy (GO) of the core target was analyzed by David database, and then the related pathways of the core target were enriched by KEGG database. RESULTS A total of 54 GO enrichment processes were obtained by GO enrichment anal?ysis of 44 common genes, including 38 biological processes (BP), 15 cell composition (CC) processes, and 1 molecular functional (MF) process. 43 items were obtained by signal pathway enrichment analysis in KEGG database. CONCLU?SION It is suggested that the mechanism of Hyp may be related to PI3K-Akt, RAP1, RAS, VEGF and other signal trans?duction pathways. The above results laid a theoretical foundation for the study of the mechanism and clinical application of the treatment of ischemia/reperfusion injury.

AIMTo develop a HPLC-ESI-MS assay for determination of eperisone hydrochloride in human plasma and investigate the pharmacokinetics and bioequivalence of two eperisone hydrochloride tablets in human.

METHODSBuflomedil hydrochloride was used as the internal standard. After alkalized with saturated sodium bicarbonate solution, plasma was extracted with diethylether-cyclohexane (1:1) and separated using HPLC on a reversed-phase C18 column with a mobile phase of 10 mmol x L(-1) ammonium acetate buffer solution (adjusted to pH 3.88 with acetic acid)-methanol (20:80). HPLC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at m/z 260 for eperisone and m/z 308 for the internal standard. A randomized crossover design was performed in 20 healthy volunteers. In the two study periods, a single 100 mg dose of each tablet was administered to each volunteer.

RESULTSCalibration curve was linear over the range of 0.02-20 microg x L(-1). The limit of quantification for eperisone hydrochloride in plasma was 0.02 microg x L(-1). The main pharmacokinetics parameters T1/2, Tmax and Cmax were (2.7 +/- 0.4) h, (1.1 +/- 0.5) h and (2.8 +/- 2.8) microg x L(-1) for the reference tablet; (2.8 +/- 0.5) h, (1.1 +/- 0.4) h and (3 +/- 4) microg x L(-1) for the test tablet, respectively. The relative bioavalability of the test tablet was (101 +/- 13)%.

CONCLUSIONThe assay was proved to be sensitive, accurate and convenient. The two formulations were bioequivalent.

Adult ; Chromatography, High Pressure Liquid ; Humans ; Male ; Propiophenones ; administration & dosage ; pharmacokinetics ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; Therapeutic Equivalency

OBJECTIVETo study the epdimiology characteristics and the diversity of VP6 gene of GCRV in Lulong, and to provide the basis for GCRV in-depth research.

METHODS793 stool specimens from porcine with diarrhea or not from Lulong in 2007 and 2008. GCRV was detected by nested multiple reverse transcription- polymerase chain reaction (nested RT-PCR) , and analyzed the identity and conducted phylogenetic tree by the seqences.

RESULTSThe positive rate of GCRV was 16.65%. Porcine GCRV strains of Lulong had significant homology differences. Phylogenetic analysis indicated porcine GCRVs were with significant diversity. Amino acid analysis showed GCRV strains with the same host shared the nearest kinship.

CONCLUSIONThe infection rate of GCRV was high from 2007 to 2008 in Lulong. Homology and phylogenetic analysis showed that VP6 gene diversity was widespread. The experimental data provided basis for molecular characteristics of porcine GCRVs.

Animals ; Antigens, Viral ; genetics ; Capsid Proteins ; genetics ; China ; Genetic Variation ; Humans ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Swine

OBJECTIVE To explore the effect of total flavonoids of Rhododendra simsii (TFR) on improving cerebral ischemia/reperfusion injury (CIRI) and its relationship with STIM/Orai-regulated operational Ca2+influx (SOCE) pathway. METHODS Oxygen-glucose deprivation/reoxygenation (OGD/R) PC12 cells were used to simulate CIRI in vitro, and the intracellular Ca2+ concentration and apoptosis rate of PC12 cells were detected by laser confocal microscope and flow cytometry, respectively. The regulation of STIM/Orai on SOCE was analyzed by STIM/Orai gene silencing and STIM/Orai gene overexpression. The CIRI model was established by MCAO in SD rats. The activities of inflammatory cyto?kines IL-1, IL-6 and TNF-αin serum were detected by ELISA. The pathological changes of ischemic brain tissue and the infarction of rat brain tissue were detected by HE staining and TTC staining. The protein and mRNA expression levels of STIM1, STIM2, Orai1, caspase-3 and PKB in brain tissue were detected by Western blotting and RT-qPCR, respectively. RESULTS The results of in vitro experiment showed that the fluorescence intensity of Ca2+ and apoptosis rate in PC12 cells treated with TFR were significantly lower than those in OGD/R group, and this trend was enhanced by SOCE antagonist 2-APB. STIM1/STIM2/Orai1 gene silencing significantly reduced apoptosis and Ca2+overload in OGD/R model, while TFR combined with overexpression of STIM1/STIM2/Orai1 aggravated apoptosis and Ca2+overload. In the in vivo experiment, TFR significantly reduced the brain histopathological damage, infarction of brain tissue, the contents of IL-1, IL-6 and TNF-α in the serum in MCAO rats and down-regulated the expression of STIM1, STIM2, Orai1 and caspase-3 protein and mRNA in the brain tissue, and up-regulated the expression of PKB. The above effects were enhanced by the addition of 2-APB. CONCLUSION The above results indicate that TFR may reduce the contents of inflammatory factors and apoptosis, decrease Ca2+ overload and ameliorate brain injury by inhibiting SOCE pathway mediated by STIM and Orai, suggesting that it has a protective effect against subacute CIRI.

This study was purposed to investigate the effect of adenovirus-mediated transfer of PDCD5 gene on apoptosis of K562 cells induced by etoposide. Recombinant adenovirus PDCD5 (Ad-PDCD5), control vectors Ad-null and Ad-eGFP were constructed by AdMax vector system respectively. After K562 cells were transfected by Ad-PDCD5, Ad-null or Ad-eGFP with different multiplicity of infection (MOI), the expression level of the PDCD5 gene was examined by RQ-RT-PCR assay. The effects of etoposide in combination with Ad-PDCD5 on the proliferation and apoptosis of K562 cells were measured by using MTT assay and flow cytometry with Annexin-V-FITC/PI dual labeling technique, respectively. The results showed that the transfection efficiencies of Ad-eGFP in K562, Jurkat and CEM cells ranged from 60% to 86%. Expression level of PDCD5 gene in K562 cells was evidently increased following transfection with Ad-PDCD5. The Ad-PDCD5 synergistically enhanced the apoptotic percentage of K562 cells induced by VP-16, as compared with that of Ad-null + VP16 and VP-16 alone respectively. It is concluded that Ad-PDCD5 may be a potential agent for enhancing the chemotherapy effect.
Adenoviridae ; genetics ; metabolism ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Etoposide ; pharmacology ; Humans ; K562 Cells ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Transfection

OBJECTIVETo investigate the ratio of Th17 cells and CD4⁺CD25⁺Foxp3⁺ regulatory T (Treg) cells in peripheral blood from patients with multiple myeloma (MM) and explore its pathological effects.

METHODS70 MM patients were divided into three groups: newly diagnosed group (n=30), plateau stage group (n=23) and relapsed/refractory group (n=17). The controls consisted of 20 healthy donors. The frequencies of Th17 and Treg cells were detected by flow cytometry.

RESULTSCompared with controls [(0.72±0.33)%] and plateau stage group [(0.74±0.29)%], frequencies of Th17 cells were higher in newly diagnosed group [(1.62±0.65)%] and relapsed/refractory group [(1.45±0.51)%], respectively (P<0.05). Compared with controls [(2.33±0.90)%] and plateau stage group [(1.69±0.70)%], frequencies of Treg cells were significantly lower in newly diagnosed group [(0.55±0.23)%] and relapsed/refractory group [(0.82±0.54)%], respectively (P<0.05). The ratios of Th17/Treg in newly diagnosed group and relapsed/refractory group were higher than those in controls (P<0.05). There were no differences of the frequencies of CD3⁺CD4⁺ T cells and Th17 cells between plateau stage group and controls. The frequencies of Treg cells were significantly lower in plateau stage group than that in controls (P<0.05), and the ratio of Th17/Treg was significantly higher in plateau stage group than that in controls (P<0.05).

CONCLUSIONThe remarkable abnormality of T cells subsets was reduction of CD4⁺ T cells in MM. Higher frequency of Th17 and lower ratio of Treg could lead to imbalance of Th17/Treg, which may play a critical role in the pathogenesis of MM.

Adult ; Aged ; Female ; Flow Cytometry ; Humans ; Lymphocyte Count ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; pathology ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology

OBJECTIVETo analyze the immunophenotype and leukemia associated immunophenotype (LAIP) of leukemia cells from patients with acute myeloid leukemia (AML) in minimal residual disease (MRD) detection.

METHODSFour-color multiparametric flow cytometry (FCM) with CD45/SSC gating was used to determine the expression of the following antibodies of CD7, CD117, CD33, CD34, CD10, CD19, CD56, CD38, CD13, CD14, CD64, CD9, CD16, CD2, CD5, CD11b, CD123, HLA-DR in 610 AML patients and 20 normal bone marrow (NBM) samples.

RESULTSThe mean percentages of CD34+ and CD117+ side scatter low (SSC(low)) cells in NBM mononuclear cells (BMMNCs) were (0.35 +/- 0.15)% and (0.76 +/- 0.31)%, respectively. The majority (84% -95%) of CD34+ SSC(low) cells co-expressed CD13, CD33, CD38, CD117 and HLA-DR. 33% and 20% of CD34+ SSC(low) cells were CD15+ and CD9, respectively. Only a small proportion (< 10%) of CD34 SSC(low) cell co-expressed CD11b, CD56, CD19 and CD64, while co-expressed CD7 was 12%. In CD117 SSC(low) cells, the relative proportions of CD19+, CD11b+, CD56+ and CD7+ were less than 10%, while CD9+ was 14%. CD38+ and HLA-DR+ were 87% and 91%, respectively. The expressions of CD15, CD34, CD13 and CD33 on CD117 SSC(low) cells were between 30% and 50%. In AML patients, most cases were CD117+ (95.08%), CD38+ (94.74%), CD9+ (84.93%), CD33+ (84.60%), HLA-DR+ (77.23%) and CD13 (75.25%). The proportions of CD64+ and CD34+ were 64.41% and 59.51%, and that of CD15 and CD11b+ were 43.06% and 22.07%, respectively. 86.39% of AML patients were found to have at least one LAIP, the highest incidence being in AML-M1 and M3 subtypes and the lowest in AML-M4Eo subtype. The cross-lineage antigen and asynchronous antigen expression were the most frequent aberrant phenotypes. CD7, CD19 and CD56 expressing on CD34+ cells were major cross-lineage antigen. For asynchronous antigen expression, CD34+ CD64+, CD117+ CD11b+ , CD34+ CD38(-/dim) and CD34+ HLA-DR(-/dim) were seldom expressed on normal BMMNCs (about 0.01%), and the logarithm difference between AML and NBM was larger than 3.0, being the more sensitive LAIP.

CONCLUSIONMRD detection by multiparameter flow cytometry can be applied to more than 80% of AML patients.

Adult ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; immunology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis

OBJECTIVETo evaluate efficacy of closed reduction and percutaneous hollow screw fixation with prototypical retractor in treating calcaneal fracture.

METHODSFrom January 2009 to June 2011, 39 patients (43 feet) with calcaneal fracture were treated by closed reduction and percutaneous hollow screw fixation with prototypical retractor. There were 33 males and 6 females, aged from 19 to 61 years with an average of 36.7 years. According to type of Sanders, type II were in 19 feet, type III were in 24 feet. Preoperative and postoperative X-ray were estimated, the data of height, width, Bölher angle, Gissane angle of calcaneous were collected, and ankle function were estimated according to AOFAS system.

RESULTSAll patients were followed up from 6 to 36 months with an average of (15.4 +/- 3.1) months. All wounds were healed well, no skin edge necrosis and infections occurred. Before operation, the height of calcaneous was average of (32.45 +/- 3.51) mm, width was (41.60 +/- 2.42) mm, Bölher angle was (8.64 +/- 13.2) degrees and Gissan angle was (136.35 +/- 15.23) degrees; while after operation, the height of calcaneous was average of (43.62 +/- 1.02) mm, width was (38.02 +/- 1.28) mm, Bölher angle was (26.87 +/- 5.32) degrees and Gissan angle was (120.78 +/- 5.34) degrees, and had significanty differences between preoperative and postoperative treatment (P<0.05). AOFAS score was improved from preoperative (35.64 +/- 11.23) to postoperative (76.18 +/- 9.87), and 29 cases got excellent results, 11 good and 3 fair. COMCLUSION: Closed reduction and percutaneous hollow screw fixation auxiliary by the retractor for the treatment is a good way, which has advantages of simple operation, satisfactory reduction fixation, reliable fixation, minimally invasive, less complications and rapid recovery.

Adult ; Bone Screws ; Calcaneus ; diagnostic imaging ; injuries ; surgery ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Fractures, Bone ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Skin ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult