Objective To evaluated the effect of hyperbaric oxygenation therapy(HBO)on the RhoA ex- pression and nerve function after transient focal cerebral ischemia in a rat model of middle cerebral artery occlusion. Methods One hundred and twenty-six healthy Sprague-Dawley rats were used and randomly divided into a sham op- eration group(shame group,n=42),a treatment group(n=42),and a control group(n=42).The animal model of middle cerebral artery occlusion(MCAO)was established by using the Zea-Longa method with the animals in the treatment and the control groups,and sham operation was performed with those in the sham group.HBO was applied to the animals in the treatment group.The RhoA protein expression was observed by using immunohistochemistry technique,and the neurological function was evaluated by Bederson's scale at different time points after MCAO.Re- sults(1)Weakly positive expression of RhoA could be located in bilateral cortex and the basal ganglia in the sham group.The expression of RhoA in the treatment group and control group was increased as early as 6 hours after MCAO when compared with that of the sham group,and peaked at 48 h after MCAO and decreased after then,but was still higher than that of the shame group at 7th day to 14th day after MCAO.It was also found that the expression of RhoA of the treatment group was significantly lower than that of the control group(P

This study aimed to study the epidemiological and clinical characteristics of human bocavirus 1-4 (HBoV1-4) in children with acute diarrhea in Lanzhou and to investigate the association between HBoV and acute gastroenteritis. A total of 331 stool samples were collected from children aged under 5 years with acute diarrhea at the Department of Pediatrics, the First Hospital, Lanzhou University, between July 2012 and June 2013. Nested PCR was used to screen for HBoV and a general PCR was employed to screen other common diarrhea viruses. We found human bocavirus 1, 2, 3 and 4 in 26, 15, 7 and 1 cases, respectively. There was no specific seasonal distribution of HBoV, with infections occurring throughout the year. HBoV was mostly found in children aged between 7 and 12 months, with a mean age of 11.04 months (+/- 6.92 months), and 93.88% of affected children were aged under 2 years. Overall, 71.3% of mixed infections were mixed and the majority of other infections were caused by rotavirus. There was no statistical difference in the incidence of fever and vomiting associated with HBoV infection. A rare virus strain, HBoV4 (LZFB086), was identified, which showed highest levels of nucleotide sequence identity (99.0%) with a single Thai HBoV strain (JQ267789). No case of HBoV2B was found. In conclusion, HBoV1 was a major etiological pathogen of HBoV in pediatric cases in Lanzhou. HBoV4 was detected in feces for the first time in China. The rate of mixed infections was high and rotavirus was dominant. The data presented suggests that HBoV is not a major causative agent of gastroenteritis.
China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Feces ; virology ; Human bocavirus ; classification ; genetics ; isolation & purification ; Humans ; Infant ; Molecular Sequence Data ; Parvoviridae Infections ; epidemiology ; virology ; Phylogeny ; Seasons

OBJECTIVETo investigate the osteoblast-like characteristics of human periodontal ligament cells affected by elastic stress in vitro, and the role of corebinding factor a 1 (cbfa1) in alveolar bone formation during orthodontic tooth movement.

METHODSRat dig-labeled cbfa1 cDNA probe was prepared from SD rat osteoblasts cultured in vitro. Human periodontal ligament cells were cultured on the elastic bottom plate and stimulated by elastic stress using mechanical loading system for cultured cells in vitro. The expression of cbfa1 mRNA was detected by in situ hybridization method.

RESULTSCbfa1 mRNA express in human periodontal ligament cells stimulated by elastic stress and did not express in normal human periodontal ligament cells.

CONCLUSIONIt is suggested that elastic stress plays a role in the differentiation process from human periodontal ligament cells to osteoblast-like cells. Cbfa1 is a transcription factor in alveolar bone remodeling during orthodontic tooth movement.

Adolescent ; Animals ; Cells, Cultured ; Child ; Core Binding Factor alpha Subunits ; DNA-Binding Proteins ; genetics ; Elasticity ; Humans ; Periodontal Ligament ; cytology ; metabolism ; RNA, Messenger ; analysis ; Rats ; Transcription Factors ; genetics

AIMTo investigate the effect of puerarin on expressions of MMP-2 and TIMP-2 in the kidney of diabetic rats.

METHODSUninephrectomized male Wistar rats were used to induce diabetes by intraperitoneal injection of streptozocin (65 mg x kg(-1)). Puerarin was given daily by intraperitoneal injection from the third day of induction of diabetes for 16 weeks. Using in situ hybridization and immunohistochemistry to detect MMP-2, TIMP-2 mRNA expressions and MMP-2, TIMP-2, collagen IV and Laminin expressions in diabetic kidneys with image analysis system, Flow cytometry was used to detect the expressions of TGFbeta1, MMP-2 and TIMP-2.

RESULTSCompared with those in kidneys of control group, expressions of MMP-2 mRNA and proteins were lower, while the expressions of both TGFbeta1 and TIMP-2 were higher in the diabetic kidney (P < 0.05). The level of MMP-2 expression was advanced, while expression of TIMP-2 was reduced by puerarin treatment (P < 0.05).

CONCLUSIONPuerarin showed some renal protective effect on diabetic nephropathy, partly through inhibition of excessive deposition of glomeruli extracellular matrix by up-regulating MMP-2 and down-regulating TIMP-2 expressions besides reducing the blood glucose.

Animals ; Collagen Type IV ; metabolism ; Diabetic Nephropathies ; chemically induced ; metabolism ; physiopathology ; Isoflavones ; pharmacology ; Kidney ; enzymology ; metabolism ; pathology ; Laminin ; metabolism ; Male ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; Peptide Fragments ; metabolism ; Protective Agents ; pharmacology ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Wistar ; Streptozocin ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1

OBJECTIVETo detect the expression profile of NK ligands in acute leukemia cell lines and investigate the differential expression pattern between acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML).

METHODSUsing quantitative real-time PCR, 23 NK ligands (MICA, MICB, ULBP-1, ULBP-2, ULBP-3, ULBP-4, HLA-E, HLA-G, CD48, NBTA, HLA-F, LLT-1, PVR, Nectin2, CD72, CD80, ICAM-1, LFA-3, CRACC, Fas, DR4, DR5, TNFR1) were detected in 6 acute leukemia cell lines, including 3 ALL cell lines (CEM, Jurkat T, Reh) and 3 AML cell lines (HL-60, KG-1a, NB4), respectively. Independent-samples t test analysis was performed to determine statistical significance.

RESULTSUsing β-actin as reference gene, the relative expression results showed that the expression of 4 NK ligands between ALL and AML is significantly different. Specifically, the level of ULBP-2 is higher in ALL (CEM: 1, Jurkat T: 0.617, Reh: 0.246) than that in AML (HL-60: 0.000, KG-1a: 0.003, NB4: 0.000)(P = 0.047). However, the expressions of CD48, PVR(PVR-1, PVR-2) and DR4 is higher in AML (HL-60: 13.987, 4.403, 10.334, 8.711; KG-1a: 5.387, 2.900, 7.315, 4.512; NB4: 7.763, 3.248, 7.049, 6.127) than that in ALL (CEM: 1, 1, 1, 1; Jurkat T: 2.035, 1.553, 3.888, 0.449; Reh: 1.559, 0.000, 0.000, 1.304) (P = 0.044, 0.014, 0.014, 0.011). And there're no significant differences between the rest 19 NK ligands.

CONCLUSIONSULBP-2, CD48, PVR and DR4 might play an important role in the distinct mechanisms in leukemogenesis between ALL and AML and could be potential targets for diagnosis and treatment.

Acute Disease ; Antigens, CD ; genetics ; metabolism ; CD48 Antigen ; Cell Line, Tumor ; GPI-Linked Proteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Ligands ; Membrane Proteins ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Receptors, Virus ; genetics ; metabolism

BACKGROUNDThe proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood (PB-MSCs) were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the feasibility for autologous PB-MSCs applications in cartilage repair.

METHODSMSCs were mobilized into peripheral blood by granulocyte colony stimulating factor (G-CSF) and AMD3100. The blood samples were collected from central ear artery of rabbits. Adhered cells were obtained by erythrocyte lysis buffer and identified as MSCs by adherence to plastic, spindle shaped morphology, specific surface markers, differentiation abilities into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. MSCs were cultured in four groups at different oxygen tension (20% O2 and 2% O2), with or without 10% fetal bovine serum (FBS) conditions: 20% O2 and 10% FBS complete medium (normal medium, N), 20% O2 and serum deprivation medium (D), 2% O2 and 10% FBS complete medium (hypoxia, H), 2% O2 and serum deprivation (HD). Cell proliferation was determined by CCK-8 assay. Apoptosis was detected by Annexin V/PI and terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL) staining.

RESULTSSpindle-shaped adherent cells were effectively mobilized from peripheral blood by a combined administration of G-CSF plus AMD3100. These cells showed typical fibroblast-like phenotype similar to MSCs from bone marrow (BM-MSCs), and expressed a high level of typical MSCs markers CD29 and CD44, but lacked in the expression of hematopoietic markers CD45 and major histocompatibility complex Class II (MHC II). They could also differentiate into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. No significant morphological differences were found among the four groups. It was found that hypoxia could enhance proliferation of PB-MSCs regardless of serum concentration, but serum deprivation inhibited proliferation at the later stage of culture. Apart from that, hypoxia or serum deprivation could promote the apoptosis of PB-MSCs after 48 hours; the effect was stronger when these two conditions combined together. Furthermore, the effect of serum deprivation on apoptosis was stronger compared with that of hypoxia.

CONCLUSIONSPB-MSCs possess similar phenotypes as BM-MSCs. Their differentiation and proliferation abilities make them a new source of seed cells for ischemia-related cell therapy and tissue engineering in the field of the articular cartilage repair.

Animals ; Apoptosis ; physiology ; Cell Hypoxia ; physiology ; Cell Proliferation ; Cells, Cultured ; In Situ Nick-End Labeling ; Mesenchymal Stromal Cells ; cytology ; Rabbits

In this study, cepharanthine hydrochloride (CH) was tested for its potential ability to modulate the expression and function of P-glycoprotein (P-gp) in the multidrug-resistant human chronic myelogenous leukemia cell line K562/ADR. Cytotoxicity of adriamycin (ADR) alone or in combination with CH or verapamil (VER) in K562 and K562/ADR cells was determined by MTT assay. Based on flow cytometric technology, the effect of CH or VER on the uptake and efflux of rhodamine123 (Rho123) and the accumulation of ADR in these cells was detected by measuring Rho123 or ADR-associated mean fluorescence intensity (MFI). The effects of CH and VER on P-glycoprotein (P-gp) expression in K562 and K562/ADR cells were also measured using a flow cytometry with PE-conjugated P-glycoprotein antibody. The results show that CH significantly enhanced the sensitivity of K562/ADR cells to ADR, 4 micromol x L(-1) of CH enhanced the sensitivity of K562/ADR cells to ADR by 7.43 folds, the reversal activity was 3.19 times higher than that of verapamil. However, CH had no effect on drug-sensitive K562 cells (P < 0.05). CH increased Rho123 and ADR accumulation in a concentration-dependent manner (2-8 micromol x L(-1)) and inhibited the efflux of Rho123 from these cells, but did not affect the accumulation and efflux of Rho123 from the wild-type drug-sensitive K562 cells. The inhibition effect of CH on P-gp expression in K562/ADR cells is in a time- and concentration-dependent manner. The reversal activity of CH is possibility related to inhibition of P-gp function and expression, which lead to an increased intracellular accumulation of anticancer drugs.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antibiotics, Antineoplastic ; metabolism ; pharmacology ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Benzylisoquinolines ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Doxorubicin ; metabolism ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; K562 Cells ; Rhodamine 123 ; metabolism

OBJECTIVETo investigate the predictive value of molecular subtypes and the evaluational value of dynamic contrast-enhanced MRI of neoadjuvant chemotherapy for breast cancer.

METHODSFrom January 2010 to December 2011, the 79 patients diagnosed as primary invasive breast cancer, having received 6 cycles of neoadjuvant chemotherapy and finished the mastectomy or the breast conserving surgery entered this study. A total of 79 patients participated in this prospective study. There were 6 (7.6%) luminal A cases, 42 (53.2%) luminal B cases, 14 HER-2 (17.7%) positive cases and 17 (21.5%) triple negative cases. The associations between molecular subtypes and clinical response as well as the pathological response were analyzed. The predictive value of molecular subtypes for the neoadjuvant chemotherapy was studied.

RESULTSClinical effective rate was 85.3% (66/79). There was no statistical correlation between molecular subtypes and clinical effective rate. Pathologic effective rate was 79.7% (63/79). There was no statistical correlation between molecular subtypes and pathologic effective rate. Twenty-seven case achieved pathologic complete remission (pCR) in all the patients. No case achieved pCR in the patients classified as Luminal A. Twelve cases (28.6%, 12/42) achieved pCR in the luminal B patients.Five cases (5/14) achieved pCR in the HER-2 overexpression patients. Ten cases (10/17) achieved pCR in the triple-negative patients. There was a statistical correlation between the molecular subtypes and the pCR rate (P = 0.039), and between clinical evaluation by dynamic contrast-enhanced MRI and evaluation of pathological response (r = 0.432, P = 0.000).

CONCLUSIONSMolecular subtypes and dynamic contrast-enhanced MRI have a good value of predicting and evaluating the response of neoadjuvant chemotherapy on breast cancer.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; pathology ; therapy ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Middle Aged ; Neoadjuvant Therapy ; Prospective Studies ; Treatment Outcome

BACKGROUNDUse of neoadjuvant chemotherapy necessitates assessment of response to cytotoxic drugs. The aim of this research was to investigate the effectiveness of dynamic contrast-enhanced magnetic resonance imaging (MRI) for evaluating clinical responses to neoadjuvant chemotherapy in breast cancer patients.

METHODSWe examined patients receiving neoadjuvant chemotherapy for primary breast cancer between October 2007 and September 2008. Dynamic contrast-enhanced MRI was used to examine breast tumors prior to and after neoadjuvant chemotherapy. The MRI examination assessed tumors using Response Evaluation Criteria in Solid Tumors (RECIST). The Miller-Payne grading system was used as a histopathological examination to assess the effect of the treatment. We examined the relationship between the results of RECIST and histopathological criteria. In addition, we used time-signal intensity curves (MRI T-SI) to further evaluate the effects of neoadjuvant chemotherapy on tumor response.

RESULTSMRI examination of patients completing four three-week anthracycline-taxanes chemotherapy treatment revealed that no patients had complete responses (CR), 58 patients had partial responses (PR), 29 patients had stable disease (SD), and four with progressive disease (PD). The effectiveness of neoadjuvant chemotherapy (CR + PR) was 63.7% (58/91). The postoperative histopathological evaluations revealed the following: seven G5 (pCR) cases (7.7%), 39 G4 cases (42.9%), 16 G3 cases (17.6%), 23 G2 cases (25.3%), and six G1 cases (6.6%). The effectiveness (G5 + G4 + G3) was 68.1% (62/91). MRI T-SI standards classified 53 responding cases, 29 stable cases, and nine progressing cases. These results indicated that the treatment was 58.2% effective (53/91) overall.

CONCLUSIONSDynamic contrast-enhanced MRI and histopathological standards were highly correlated. Importantly, MRI T-SI evaluation was found to be useful in assessing the clinical effectiveness of neoadjuvant chemotherapy.

Adult ; Aged ; Anthracyclines ; therapeutic use ; Antineoplastic Agents ; therapeutic use ; Breast Neoplasms ; drug therapy ; pathology ; Contrast Media ; chemistry ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Middle Aged ; Neoadjuvant Therapy ; Taxoids ; therapeutic use

Objective To study the clinical and molecular epidemiology characteristics of human Bocavirus 1-3 (HBoV1-3) in children for acute respiratory infection in Lanzhou area. Methods Nasopharyngeal aspiration samples and throat swabs were collected from 524 children with ARTI at the First Hospital of Lanzhou University,Gansu Province,China,between December 2009 and November 2010. Nested PCR was employed to screening HBoV1-3, which amplified a 518-bp fragment of the partial NS1 gene.Furthermore, a standard reverse transcription-PCR was used to screen for other common respiratory viruses.Results The overall frequency of H BoV was 8.2% (43/524), lining up behind human rhinovirus, RSV,parainfluenza virus 3. Thirty of the HBoV-postive children (69. 8% ) were co-infected with other respiratory viruses. The prevalence of HBoV1 in ALRTI was obviously higher than that in AURI. The 2 HBoV2 NS1 sequences shared 99% and 100% nucleotide sequence identity with HBoV2 strain CU47TH respectively.Two cases of HBoV2 postive children appears gastrointestinal symptoms. The one HBoV3 NS1 sequences shared 99% nucleotide sequence identity with HBoV3 isolate 46-BJ07. Conclusion The HBoV3 was detected at the first time in lanzhou area. HBoV1-3 infection exists in children with acute respiratory tract infections in Lanzhou region, HBoV1 were dominant. The mixed infection rate was higher.