Objective To isolate and culture sweat gland epithelial cells in vitro,and to study the effects of acetylcholine (ACh) on intracellular flee calcium concentration ([Ca~(2+)]i) of cultured sweat gland epithelial cells.Methods Sweat glands epithelial cells were collected by enzymatic digestion.After ACh was added to the primary and first passage cells,[Ca~(2+)]i was examined using confocal laser scanning microscopy (CLSM) and the Ca~(2+) sensitive dye Fura 3/AM.Results The primary and first passage epithe- lial cells grew well.After ACh was added,opening of the calcium channel and significant [Ca~(2+)]i increase were observed when the primary and first passage cells were incubated with high concentration of calcium (2 mmol/L);no significant [Ca~(2+)]i increase was observed in those cultured without calcium.Conclusion Upon stimulation with ACh,calcium channels of cultured primary and first passage sweat gland epithelial cells would open,influx of extracellular Ca~(2+) occurred,which resulted in an increase of [Ca~(2+)]i.Extracellular bound calcium was therefore converted into intracellular free calcium.

OBJECTIVETo design a new type minor smoke warming moxibustion cup for convenient use of both the physicians and the patients.

METHODSThe double-deck minor smoke warming moxibustion cup is fixed on the part receiving moxibustion by vacuum adsorption; the filtration device on the upper can filtrate and adsorb the harmful substance in the moxa-smoke, and the device with a double-temperature control on the lower can sensitively regulate the moxibustion temperature.

RESULTSThis new type minor smoke warming moxibustion cup has the advantages of minor smoke discharge, convenient fixation, sensitive regulation of temperature, saving moxibustion material, lasting action, safety, besides the advantages of traditional moxibustion.

CONCLUSIONThe new type minor smoke warming moxibustion cup can use for treatment and prevention of diseases, suitable to clinical treatment and family health care.

Equipment Design ; Humans ; Moxibustion ; instrumentation

OBJECTIVETo evaluate the usefulness of new microspincolumn method for the measurement of a1pha-fetoprotein variant AFP-L3 in differentiation of benign and malignant liver disease and the warming for liver cancer.

METHODSAFP-L3 was isolated by using microspincolumn coupled with lens culinaris agglutinin (LCA), AFP and AFP-L3 were determined with chemiluminescent immunoassay, the proportion of AFP-L3 levels AFP-L3(%) were calculated, and the relationship between the elevated AFP-L3(%) levels and benign and malignant liver disease was analyzed.

RESULTSThe levels of AFP-L3(%) in serum of patients with hepatocellular carcinoma was significantly higher than those in the patients with other liver diseases (P < 0.001). Taking AFP-L3(%) >or= 10% as the diagnostic criteria, the sensitivity for diagnosis of liver cancer was 90.9%.

CONCLUSIONDetection of AFP-L3 seemed to be of clinical value in diagnosis and differential diagnosis of hepatocellular carcinoma; it may be especially important for identifying patients with hepatocellular carcinoma whose a1pha-fetoprotein level is low.

Adult ; Aged ; Carcinoma, Hepatocellular ; blood ; diagnosis ; Diagnosis, Differential ; Female ; Hepatitis, Chronic ; blood ; diagnosis ; Humans ; Immunoassay ; methods ; Liver Cirrhosis ; blood ; diagnosis ; Liver Neoplasms ; blood ; diagnosis ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; analysis

OBJECTIVETo study the role of adaptor protein Dok6 in neurite outgrowth in PC12 cells.

METHODSSeries of fusion clones were constructed by fusing different domains of Dok6 into mutant TrkC/Y516F. These constructs were transiently transfected into PC12 cells separately and the expression levels of fusion proteins were detected by Western blot. Neurite outgrowth in these PC12 cells was tested after stimulation of NT-3.

RESULTSEach fusion clone was stably expressed in PC12 cells. The fusion clones that fused both TrkC/Y516F-Dok6 (PTB+C) and TrkC/Y516F-Dok6C rescued the loss of neurite outgrowth in PC12 cells resulting from the mutation in tyrosine 516, while fusion clones that fused with single TrkC/Y516F-Dok6PTB did not show such effect.

CONCLUSIONDok6 can promote neurite outgrowth induced by NT-3 stimulation through its C-terminal in TrkC-positive PC12 cells.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Animals ; Neurites ; drug effects ; physiology ; Neurotrophin 3 ; pharmacology ; PC12 Cells ; Rats ; Receptor, trkC ; metabolism ; Transfection

OBJECTIVETo study the interaction between ShcD and TrkC and to reveal the molecular mechanism of the downstream signal transduction of TrkC.

METHODSYeast two-hybrid assay was used. The intracellular domains of TrkC and TrkC mutants were cloned into pAS2-1, and ShcD and its four domains (CH2, PTB, CH1, and SH2 domains) were cloned into pACT2 vector respectively. The constructs were separately cotransformed into yeast. beta-galactosidase activity was measured to detect their interactions. TrkC was cloned into pmRFP (carrying red fluorescent protein), and ShcD was cloned into pEGFP (carrying green fluorescent protein). pmRFP-TrkC and pEGFP-ShcD were co-transfected into 293T cells, and then the cells were fixed and subjected to confocal analysis to study their subcellular localization.

RESULTSShcD interacted with TrkC but not with kinase dead mutant TrkCM1(K572A). Both PTB and SH2 domains were capable of binding to TrkC, and PTB domain bound NPQY motif of TrkC. ShcD colocalized with TrkC throughout the cytoplasm and in the plasma membrane in 293T cells.

CONCLUSIONShcD binds to TrkC in a kinase-activity-dependent manner through its PTB and SH2 domains.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Binding Sites ; Cells, Cultured ; Genetic Vectors ; Humans ; Plasmids ; genetics ; Protein Binding ; Receptor, trkC ; genetics ; metabolism ; Shc Signaling Adaptor Proteins ; genetics ; metabolism ; Transfection ; Transformation, Bacterial ; Two-Hybrid System Techniques ; src Homology Domains ; genetics

Objective: The research is to provide the basis for effective protection and wise utilization of the plants resources inTea Mountain National Ecological Demonstration Area of Shandong Province. Methods: The general investigation on plant species inTea Mountain was conducted. The research scope covered plant species, their distribution in wild, community structure and flora.Results: A total of 352 higher plants, belonging to 250 genera and 90 families, were recorded. Four species were listed as nationalsecond-grade protected plants. Regional vegetation is Pinus thunbergii plantation and Secondary forest. The main forest vegetation isPinus thunbergii community. Pantropic types and N. Tem types are dominant types. The latitude and "the islands-effect" should be themajor-effect factor of this pattern. Because of intense artificial active disturbance, primary vegetation types in this region have sufferedserious destruction, especially the rare and precious plants that are at high risk of dying out. Conclusions: This regional communitystructure is simple. The shrub layer and the liana layer are undeveloped. The syngenesis time is short.

Objective To determine the prevalence and distribution of type 2 diabetes mellitus (T2DM) and the relationship between maximum body mass index (MAXBMI) and T2DM. Methods From June to August, 2005, a stratified cluster sampling of 1071 permanent residents in communities, over 20 years old, from 4 districts and 1 county of Mudanjiang was chosen. The prevalence of T2DM, and the association between T2DM and different levels of the MAXBMI, current BMI were studied. Results The prevalence in the communities was 7.09% and in those with past maximum BMI≥28 kg/m~2, it was 12.10%. With the increase of past MAXBMI levels, the risk of T2DM patients also increased significantly(trend X~2=17.387 23, P<0.0001). Data from multifactor analysis showed that MAXBMI in the past was positively related to T2DM (OR=3.06, P=0.0013). In T2DM patients, the group with MAXBMI≥27.4 kg/m~2 had higher 2-hour postprandial blood glucose than those with lower MAXBMI (P=0.0408). When compared with low maximum BMI group in normal blood glucose population, the group with higher MAXBMI (≥ 25.4 kg/m~2) had higher blood glucose and greater change of BMI. Conclusion In both groups that patients with T2DM and with normal glucose, in order to control blood glucose better, researchers should not only concern about the influence of the MAXBMI in the past, but also pay attention to constantly keep BMI at the normal range.

The cDNA fragments of interest were amplified using Sj lambda ZipLox library as the templates by PCR and then cloned into a eukaryotic expression vector p-CMV-GH; A small number of DNA fragments inserted in the recombinants was identified by restriction cleavage, EST sequencing and bioinformatical analysis; mice were injected intramuscularly with the expression library (L-CMV-SjR) or sublibraries(L-CMV-SjR1, L-CMV-SjR2 and L-CMV-SjR3), immunized mice were challenged with Schistosoma japonicum cercariae on day 35, the levels of IgG antibodies in sera from the immunized mice were detected by ELISA. The results demonstrated that a partial cDNA expression library of S.j, with approximately 10(5) transformants, was constructed, most of the recombinants contained the insert DNA fragments of interest, and these fragments had the features of protein-coding sequences for Schistosome. There were no significant differences for the levels of IgG antibodies in sera from all of the immunized groups. Mice immunized with L-CMV-SjR, L-CMV-SjR1 and L-CMV-SjR2 developed significant protective effect against Sj infection compared to control mice injected with the empty plasmid, the rate of worm reduction was about 30%.
Animals ; Antibodies, Helminth ; blood ; immunology ; Antigens, Helminth ; genetics ; immunology ; DNA, Complementary ; Disease Models, Animal ; Gene Expression ; Gene Library ; Mice ; Schistosoma japonicum ; genetics ; immunology ; Schistosomiasis japonica ; prevention & control ; Vaccination ; methods ; Vaccines, DNA ; genetics ; immunology

OBJECTIVETo screen low molecular weight protein biomarkers relevant to portal vein tumor thrombi (PVTT) in serum of hepatocellular carcinoma (HCC) patients.

METHODSSerum samples were obtained from 12 healthy volunteers, 12 HCC patients without PVTT and 12 HCC patients with PVTT. Using two-dimensional gel electrophoresis (2-DE) in which the second dimension was 16% SDS-PAGE, serum protein images of the 3 groups were analyzed by ImageMaster software. The differential protein spots were further identified by MALDI-TOF MS/MS.

RESULTSComparing the results using 12.5% SDS-PAGE gel, there were more protein bands (between 3 x 10(3) and 20 x 10(3)) and low molecular weight (MW) protein spots (less than 20 x 10(3)) were clearly shown in the 16% SDS-PAGE gel. Fifteen differential protein spots representing 5 proteins were found in the 3 groups by inter-class comparison and they were then identified. Compared with those in the healthy group, apolipoprotein A-I, lipoprotein CIII, transthyretin and DNA topoisomerase II were all down regulated in HCC groups and haptoglobin-2 was over expressed. All 5 proteins decreased more in the PVTT group than in the non-PVTT group.

CONCLUSIONThe expression of low MW serum protein obviously changes in the beginning and in the progressive stage of HCC, and differentially expressed low MW proteins might be potential biomarkers in an early prognostic prediction and surveillance in the treatment for HCC and PVTT.

Adult ; Blood Proteins ; analysis ; Carcinoma, Hepatocellular ; blood ; pathology ; Electrophoresis, Gel, Two-Dimensional ; methods ; Female ; Humans ; Liver Neoplasms ; pathology ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Portal Vein ; pathology ; Proteome ; analysis

OBJECTIVEA mouse model of orthotopic bladder cancer simulating its human counterpart is of great importance in preclinical evaluation of new treatment modalities such as immunotxin therapy. The aim of the present study is to establish a novel nude mouse model with xenografted human bladder cancer.

METHODSSingle cell suspension of an established human bladder transitional cell carcinoma (TCC) cell line BIU-87 was instilled into nude mouse bladders which were pretreated with mild acid washing. The tumor growth in mouse bladder was assessed weekly by magnetic resonance imaging (MRI). At intervals following implantation and MRI tumor detection, the animals were sacrificed for necropsy, histological examination and immunocytochemical studies.

RESULTSThe overall tumor establishment was 92.9% (52/56 mice) at 7 - 36 days, while in the subgroup of animals sacrificed at 12 - 13 days, 40 out of 42 animals (95.2%) developed TCC, the majority of which was superficial. The tumor stages were assessed by gross and histopathology. Histological examination confirmed the presence of grade II - III TCC. Immunocytochemistry confirmed that the tumor model maintained the biological and immunological features of BIU-87 cells. The changes seen on MRI images well correlated with the extent of tumor invasion identified by histology. Carcinoma in situ could be detected histologically at 7 - 9 days post-inoculation and progressed into papillary or invasive tumors thereafter.

CONCLUSIONThe orthotopic BIU-87 TCC model in nude mice is highly reproducible and is ideal for preclinical studies on experimental intravesical therapies.

Animals ; Antibodies, Monoclonal ; analysis ; Carcinoma, Transitional Cell ; immunology ; pathology ; Cell Line, Tumor ; Female ; Humans ; Immunohistochemistry ; Magnetic Resonance Imaging ; Mice ; Mice, Nude ; Neoplasm Staging ; Neoplasm Transplantation ; Neoplasms, Experimental ; pathology ; Transplantation, Heterologous ; Urinary Bladder Neoplasms ; immunology ; pathology