Objective To study the association between clinical symptoms,psychosomatic factors and autonomic nervous function in patients with gastroesophageal reflux disease (GERD).Methods Thirty-four patients with GERD diagnosed by reflux disease questionnaire (RDQ) and endoscopy and 15 healthy control subjects were enrolled in this study.All the subjects were divided into two groups,one with normal scores of Zung's self-rating anxiety scale (SAS) and Zung's depression scale (SDS) as GERD (-) and the other with abnormal scores of SAS and SDS as GERD (+).Reflux symptom score was recorded for both groups at the same time.Autonomic nervous function was assessed by their heart rate variability (HRV) on dynamic electrocardiogram (DCG).The time domain parameters analyzed included standard deviation (SD) of average R-R interval during 24 hours (SDNN),SD of average 5-minute sinus heart rate (SDANN),mean square root of the difference of adjoining two R-R interval (rMSSD),and proportion of the heart beats with difference of R-R interval more than 50 ms from the total heart beats (PNN 50),and the frequency domain parameters analyzed included low-frequency (LF),high-frequency (HF) and ratio of LF to HF.Results Average scores of SAS and SDS were significantly higher in patients with GERD than those in healthy controls (48?9 vs 38?6 and 48?11 vs 41?6,respectively,P

Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

Objective To investigate the viral prevalence of acute lower respiratory tract infection (ALRTI)in children. Methods Totally 1165 children with clinical diagnosis of ALRTI during the period from August 2007 to September 2008 were involved in our study. The nasopharyngeal aspirate specimen was collected from each patient. RT-PCRs were performed to detect common respiratory tract viruses including respiratory syncytial virus (RSV), rhinovirus (HRV), parainfluenza virus (PIV, type 1 -3 ), influzenza virus type A and B (IFA,IFB), and PCR was used to detect adenovirus (ADV). Results 783 patients were identified to have at least one kind of viral pathogens and the overall positive rate was 67.2%. The most common virus was RSV (27%), followed by HRV ( 17.4% ) and PIV3 ( 13. 9% ). The peak infection seasons were winter and autumn. The etiological spectrum of ALRTI varied in different age groups. Two or more viruses were identified in 284 out of 783 cases ( 36. 3% ). The mixed infection rate was high in infants under 1 year old (63.7%) while it decreased to 8. 5% in children older than 3 years of age. Conclusion RSV, HRV and PIV3 were the most predominant pathogens in children less than 1 year old. The peak infection seasons were winter and autumn. The infection rate and mixed infection rate in infants under 1 year old were highest. The most common style was RSV and HRV mixed infection.

OBJECTIVETo investigate the effect of piperphentonamine hydrochloride (PPTA) on cognitive deficits induced by ischemia-reperfusion and explore the possible mechanisms.

METHODSSD rats were randomly divided into sham-operated group, ischemia-reperfusion group (with saline injection), PPTA-treated groups (2.5, 5, 10 mg/kg) and edaravone-treated group (6 mg/kg). Cerebral ischemia-reperfusion injury was induced by middle cerebral artery occlusion, and the agents were administrated 1 h after ischemia. At 24 h after ischemia, step-through passive avoidance test was carried out, and 24 h later IL-1β, TNF-α, caspase-3 and HSP-70 mRNA expressions in the ischemic brain tissues were measured with RT-PCR.

RESULTSIn the step-through passive avoidance test, the rats in the ischemia-reperfusion group showed significantly shorter latency and more error times than those in the sham group, and these behavioral changes were improved significantly by treatments with PPTA and edaravone. Cerebral ischemia-reperfusion caused significantly increased expressions of IL-1β, TNF-α, caspase-3 and HSP-70 mRNA, and these changes were obviously reversed by PPTA, but not by edaravone.

CONCLUSIONSPPTA can reverse cognitive deficits induced by cerebral ischemia-reperfusion probably by decreasing the inflammatory responses and cell apoptosis in the brain, suggesting its potential as a new therapeutic agent for improving the cognitive function following cerebral ischemia-reperfusion.

3,4-Methylenedioxyamphetamine ; analogs & derivatives ; therapeutic use ; Animals ; Brain Ischemia ; drug therapy ; Cognition Disorders ; prevention & control ; Infarction, Middle Cerebral Artery ; drug therapy ; Male ; Neuroprotective Agents ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; prevention & control

Adenovirus type 40 and 41 (Ad40, Ad41), which belong to human adenovirus subgroup F, are called fastidious adenoviruses due to their property of poor growth in cultured cell lines in vitro The effect of expression of exogenous E1B55K in Hep2 on Ad41 replication in this cell line was investigated. E1B55K gene was amplified by PCR with DNA extracted from Ad41-positive feces supernatant as template. Eukaryotic expression plasmid (pcDNA3) carrying E1B55K was constructed, purified, and transferred into Hep2 cell. Expression of E1B55K in G418-resistant clones was assayed by RT-PCR, and one clone named as Hep2-E1B4#4 could produce more Ad41 progenies when compared with other clones by the method of inducing complete cytopathic effect (CPE) in 293 cells. Infection of equivalent Ad41 caused more significant cytopathic effect (CPE) in Hep2-E1B#4 than that in the control cells of Hep2 or Hep2-DNA3, also suggesting enhanced viral replication in Hep2-E1B#4. The titer of Ad41 was further determined by method of immunocytochemical staining, and semi-quantity PCR was employed to compare the copy number of Ad41 genome DNA. The results showed that the yield of Ad41 in Hep2-E1B#4 was more than 9 times of that in control cells when equal amount of seed viruses were incubated, and the copy number of Ad41 genome increased 4 times in the raw extract from the infected Hep2-E1B#4 when compared with that from control cells. In conclusion, E1B55K gene transfer improved the ability of Hep2 in packaging Ad41, and the Hep2-E1B#4 cell line, which expressed E1B55K constitutively, would be helpful in isolation, cultivation and amplification of Ad41.
Adenovirus E1B Proteins ; genetics ; metabolism ; Adenoviruses, Human ; genetics ; Cell Line, Tumor ; Gene Expression ; Humans ; Immunohistochemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Virus Replication ; genetics

The article puts forward the treatment of irritable bowel syndrome through regulating heart vitality since it is held that the pathological factors of the disease lay in dysfunction of heart and intestines as well as disorder of qi circulation. At the same time, the internal-external relationship between the heart the small intestine is discussed from the theory of Brain-gut Axis in modern medicine, which provides theoretical base of modern medicine for the treatment of irritable bowel syndrome through regulation of the heart functions.
Acupuncture Therapy ; Adult ; Female ; Heart ; physiopathology ; Humans ; Intestines ; physiopathology ; Irritable Bowel Syndrome ; physiopathology ; therapy ; Middle Aged

Objective To study the influence on the tumor after percutaneous intra-tumor injection of ~(32)P-GMS in liver cancer as well as its suitable dose.Methods 24 New Zealand rabbits were used to establish the animal model of VX-2 liver cancer,and divided into A,B,C and D groups with individually 37,74,111 and 148 MBq of ~(32)P-GMS being injected,respectively;and then pathological changes of tumor were observed by light and electron microscope respectively.Result The dose of ~(32)P-GMS was obviously correlated with the radioactivity damage of tumor cells.In the A and B groups,the tumor cells were not observed to disappear completely after injection of ~(32)P-GMS,but in C group,tumor cells were almost completely disappeared and surrounded by a lot of connective tissue.Although the tumor cells were found to disappear completely in D group,normal liver tissues were also involved.Conclusion Percutaneous intra-tumor injection of ~(32)P-GMS with suitable dose that may induce the tumor tissue to be maximally damaged and may also provide some significances to prevent the tumor metastasis.

Acute Disease ; Adult ; Aged ; Cell Differentiation ; Humans ; Leukemia ; classification ; pathology ; Male

OBJECTIVETo study the expression of ecotropic viral integration site (EVI1) gene in childhood acute myeloid leukemia (AML) and the clinical features of EVI1-positive children with AML.

METHODSThe clinical data of EVI1-positive children with AML were collected and analyzed. RT-PCR and real-time quantitative PCR were used for qualitative and quantitative analysis of expression of EVI1. Flow cytometry (FCM) was used for determining the immunophenotypes of bone marrow cells. Multiparameter FCM was used for monitoring minimal residual disease. The karyotypes were determined.

RESULTSOf 241 children with AML, 33 (13.7%) were positive for EVI1 expression. There were no significant differences in age at first visit as well as the white blood cell count, hemoglobin level, and platelet count in peripheral blood between EVI1-positive and EVI1-negative children with AML (P>0.05), but EVI1-positive children had a significantly increased proportion of females compared with EVI1-negative children (P<0.05). The change in EVI1 expression was not synchronous with clinical remission and the change of MRD: some children had clinical remission or negative conversion of MRD before negative conversion of EVI1, while some had negative conversion of EVI1 before clinical remission or while MRD showed positive. EVI1 gene was usually co-expressed with other fusion genes. CD33 (100%), CD38 (88%), and HLADR (76%) were highly expressed in EVI1-positive children with AML. Abnormal chromosome structure or number was found in 15 patients. Compared with EVI1-negative children, EVI1-positive children had significantly lower complete remission rates after the first course of treatment (P<0.05).

CONCLUSIONSEVI1-positive children with AML have a poor short-term prognosis. In the development of AML, the activation of EVI1 gene is not isolated, but the result of interactions with other genes or chromosome abnormalities, and the mechanism of activation and its function need further study.

Adolescent ; Child ; Child, Preschool ; Chromosome Aberrations ; DNA-Binding Proteins ; genetics ; Female ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Immunophenotyping ; Infant ; Leukemia, Myeloid, Acute ; genetics ; immunology ; MDS1 and EVI1 Complex Locus Protein ; Male ; Neoplasm, Residual ; Prognosis ; Proto-Oncogenes ; genetics ; Transcription Factors ; genetics

OBJECTIVETo investigate the isolation, purification and ex vivo expansion of CD34(+)CD59(+) cells from the bone marrow of children with paroxysmal nocturnal hemoglobinuria (PNH), to evaluate the capability of long-term hematopoietic reconstruction of the expanded CD34(+)CD59(+) cells, and to provide a laboratory basis for novel treatment of PNH.

METHODSCD34(+)CD59(+) cells were isolated from the bone marrow mononuclear cells of children with PNH using immunomagnetic beads and flow cytometer in sequence. The isolated cells were subjected to ex vivo expansion in the presence of different combinations of hematopoietic growth factors for two weeks. The colony-forming cells and long-term culture-initiating cells (LTC-ICs) were cultured and counted.

RESULTSThe optimal combination of hematopoietic growth factors for ex vivo expansion was stem cell factor+interleukin (IL)-3+IL-6+FLT3 ligand+thrombopoietin+ery-thropoietin, and maximum expansion (30.4 ± 6.7 folds) was seen on day 7 of days 4 to 14 of ex vivo expansion. After ex vivo expansion, CD34(+)CD59(+) cells remained CD59-positive, retained strong capability of forming colony-forming units, and could still form LTC-ICs. There was no significant difference in capability of forming LTC-ICs between CD34(+)CD59(+) cells before and after expansion. The expansion capability of CD34(+)CD59(+) cells from children with PNH was significantly lower than that of CD34(+) cells from normal controls (P<0.01).

CONCLUSIONSThe CD34(+)CD59(+) cells from children with PNH can be expanded in vitro. Post-expansion CD34(+)CD59(+) cells retain capability of long-term hematopoietic reconstruction. CD34(+)CD59(+) cells showed no trend towards PNH clone during culture. Ex vivo expansion of CD34(+)CD59(+) cells from children with PNH might be practical in performing autologous transplantation clinically for these children.

Adolescent ; Antigens, CD34 ; analysis ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; CD59 Antigens ; analysis ; Cell Separation ; Child ; Female ; Hematopoiesis ; Hemoglobinuria, Paroxysmal ; therapy ; Humans ; Male