OBJECTIVETo compare the efficacy differences between dog-days medicinal vesiculation and regular-day medicinal vesiculation for perennial allergic rhinitis (PAR), and observe their effects on serum immune globulin E (IgE) and interleukin-4 (IL-4).

METHODSSeventy-two patients were randomly divided into a dog-days moxibustion group (34 cases) and a regular-day moxibustion group (38 cases). In the dog-days moxibustion group, medicinal vesiculation was applied on the 1st dog-day, 2nd dog-day and last dog-day in summer by lunar calendar, 3 treatments per dog-day for totally 9 times. In the regular-day moxibustion group, the moxibustion was given on the regular day for continuous 9 times. The symptom score, rhinoconjunctivitis quality of life questionnaire (RQLQ) and the level of IgE and IL-4 were compared before and after treatment in two groups; the short-term and two-year efficacy evaluation were performed too.

RESULTSThe short-term total effective rate was 88.2% (30/34) in the dog-days moxibustion group, which was not significantly different to 86.8% (33/38) in the regular-day moxibustion group (P>0.05). The long-term total effective rate was 97.1% (33/34) in the dog-days moxibustion group, which was significantly superior to 81.6% (31/38) in the regular-day moxibustion group (P<0.05). After treatment, the serum IgE, IL-4 and RQLQ were significantly reduced (all P<0.01), but the difference between two groups was not significant (all P>0.05).

CONCLUSIONMedicinal moxibustion could be taken as a regular treatment for PAR, which could be performed during the whole year, and dog-days moxibustion could be considered as an enhanced method for prevention and treatment of PAR.

Acupuncture Therapy ; Adult ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Rhinitis, Allergic, Perennial ; drug therapy ; therapy ; Treatment Outcome ; Young Adult

Objective To construct PPAR?and PPAR?response element (PPRE)-controlled luciferase expression vectors,and to determine whether the traditional Chinese medicine emodin activates PPAR?and improves the glucose uptake by HepG2 hepatocytes.Methods (1) PPAR?and PPRE luciferase expression vectors were constructed and were applied to screen more than 20 ingredients of the traditional Chinese medicine. (2) HepG2 cells were incubated with emodin which can activate PPAR?and PPRE luciferase activity,and the PPAR?mRNA expression level was evaluated by RT-PCR/Southern blot.(3) PPAR?and glucose transporter 2 (Glut2) proteins were determined by Western blot analysis in HepG2 cells treated with emodin.(4) The glucose uptake rate was measured using 2-deoxy-[~3H]-D-glucose in HepG2 cells after treatment with emodin.Results (1) Emodin stimulated luciferase activity controlled by PPRE in dose-dependent manner at concentrations of 0.04 to 180?mol/L in COS-7 cells.The highest value was about 4 folds of control in the cells treated with 90?mol/L emodin (P

Objective To analyse the high-dose dexamethasone suppression test(HDDST)-related differences in the clinical and biochemical features of the patients with Cushing's disease Methods Cases were drawn from 60 consecutive patients with Cushing's disease,who were then divided into two groups according to the response to the HDDST.The clinical and biochemical features between two groups were compared.Results(1) Of the 60 patients with Cushing's disease,23.3%(14/60)of patients(group A)did not yield results of suppression with the HDDST,and the others(group B)did.No difference was found in the age[(33.8?10.4 vs 36.2?11.2)years]and duration of illness[(2.1?1.6 vs 3.9?3.1)years]between two groups.(2)In clinical features,the patients in group A were more likely to have edema of lower limbs(64.3% vs 32.6%),hypokalemia (71.4% vs 28.3%),secondary diabetes(57.1% vs 26.1%)and purple striae(85.7% vs 54.3%,all P

OBJECTIVETo investigate the methylation status of LRP15 gene in acute leukemia (AL) patients and its role in the tumorigenesis.

METHODSThe methylation of LRP15 promoter and first exon of bone marrow mononuclear cells in 73 patients with AL, 10 with chronic leukemia (CL), 9 with hematological benign diseases, and 20 healthy transplantation donors was analyzed by using methylation specific polymerase chain reaction. The methylation of LRP15 gene promoter and first exon in COS7, K562, and HL60 cell lines was also assayed.

RESULTSNo LRP15 gene promoter methylation was detected in COS7 cell line. LRP15 gene promoter was methylated in K562 and HL60 cell lines. No deletion of LRP15 gene was detected in all samples. In nearly all French-American-British leukemia subtypes, we found that frequency of LRP15 methylation in adult patients with AL was 71.23% (52/73). There was no detectable methylation in any of the 20 healthy donors and 8 chronic myeloid leukemia patients. The difference in frequency of LRP15 methylation between AL patients and healthy donors or CL patients (10.00%, 1/10) was significant (P < 0.01). Hypermethylation of LRP15 gene was found in 57.14% (16/28) of newly diagnosed AL patients, 83.33% of relapsed AL patients respectively, which was significantly different (P < 0.05). We also demonstrated LRP15 methylation in 55.56% (5/9) adults with benign hematological diseases.

CONCLUSIONSLRP15 methylation changes are common abnormalities in leukemia. LRP15 is postulated to be a tumor suppressor gene.

Acute Disease ; Animals ; Base Sequence ; COS Cells ; Cell Line, Tumor ; Cercopithecus aethiops ; DNA Methylation ; DNA Primers ; Humans ; Leukemia ; genetics ; Neoplasm Proteins ; genetics ; Promoter Regions, Genetic

This study was aimed to obtain higher efficiency in gene transfection into K562 cells and to study the role of green fluorescence protein (GFP) as a reporter system. Transfection efficiencies with different methods including electroporation and lipofectamine 2000 transfection, were observed and calculated under fluorescent microscopy by using GFP as a reporter system. The results showed that the transfection efficiency with electroporation (10%) was higher than that with lipofectamine 2000 (1%). In conclusion, the electroporation is a more ideal method for introduction of foreign gene into K562 cells. GFP can be used as a reporter system for optimizing transfection of K562 cells.
Electroporation ; Genes, Reporter ; Green Fluorescent Proteins ; genetics ; Humans ; K562 Cells ; Liposomes ; Transfection

The study was aimed to identify a new leukemia related gene zo-1 from leukemia and to explore its mechanism in leukemia. Methylation specific PCR (MSP) was used for testing gene zo-1 methylation in leukemia cells. The gene zo-1 specific siRNA was designed according to its sequence, and transfected into THP-1 cell, and the cells were cultured for 48 hours before harvesting. The effect of zo-1 siRNA was monitored by RT-PCR. The cellular proliferation activity was assayed by CCK-8, the apoptosis was detected by Annexin-V-fluorescence in isothiocyanate (FITC) assay, and cell cycle was observed by propidium iodide (PI). The results indicated that the gene zo-1 in patients with acute leukemia was hypermethylated, while the gene zo-1 in healthy persons was unmethylated. The THP-1 cells with unmethylation of zo-1 gene promoter overexpressed the gene zo-1, while the Molt4 and HL-60 cells with hypermethylation of gene zo-1 promoter did not express the gene zo-1. The silenced zo-1 gene in Molt4 and HL-60 leukemia cell lines could be reactivated by demethylation treatment with 5-AZA-dC. The oligofectamine-transfected siRNA for zo-1 gene successfully inhibited the expression of gene zo-1 in THP-1 cells, but did not interfere with cell proliferation, cell cycle and apoptosis. It is concluded that gene zo-1 is a leukemia-related gene. Gene zo-1 in acute leukemia was hypermethylated, the methylation status of gene zo-1 regulates the expression of gene zo-1. Lack of gene zo-1 expression in THP-1 cells does not influence the cell proliferation, apoptosis and cell cycle, which suggests that the methylation of gene zo-1 may be involved in the genesis of acute leukemia, its mechanism is worthy to be studied.
CpG Islands ; DNA Methylation ; Gene Silencing ; HL-60 Cells ; Humans ; Leukemia ; genetics ; Membrane Proteins ; metabolism ; Phosphoproteins ; metabolism ; RNA, Small Interfering ; genetics ; Zonula Occludens-1 Protein

OBJECTIVETo explore ZO-1 gene expression and methylation in leukemia cells and the involvement of ZO-1 gene in leukemogenesis.

METHODSRestriction landmark genomic scanning (RLGS) was used to identify new leukemia related gene, and methylation specific PCR (MSP) for ZO-1 methylation status. ZO-1 specific siRNA was designed and prepared by in vitro transcription and transfected into K562 cells, the transfected cells were cultured for 48 hours before harvesting. The effect of ZO-1 siRNA was monitored by Northern blot. Cellular proliferation capacity was assayed by CCK-8, cell apoptosis by Annexin V-fluorescence in isothiocyanate (FITC) assay, and cell cycle by phosphatidylinositol (PI).

RESULTSThe intensified spots in RLGS gel were subjected to bioinformatics analysis and one of the candidate spots was proved to be ZO-1 gene. In fresh leukemia cells, Molt4 cells and HL-60 cells, ZO-1 was hypermethylated, causing it reduced or silenced. ZO-1 gene was highly expressed with no methylation in normal peripheral blood MNC and K562 cells. There was a good correlation between promoter methylation and the gene silence. The silenced gene can be re-activated by demethylation treatment with 5-AZA-dC in most leukemia cell lines. RNA interference for ZO-1 gene in K562 cells did not interfere with cell proliferation, cell cycle and apoptosis.

CONCLUSIONZO-1 gene methylation might be involved in the tumorigenesis of acute leukemia.

Animals ; DNA Methylation ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; genetics ; pathology ; Membrane Proteins ; genetics ; metabolism ; Mice ; Phosphoproteins ; genetics ; metabolism ; Zonula Occludens-1 Protein

This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
Base Sequence ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Receptors, KIR ; biosynthesis ; genetics

The objective of this study was to investigate the methylation status of zonula occluden protein-1 (ZO-1) gene in patients with myelodysplastic syndrome (MDS) and to identify its roles in pathogenesis, development and classification of MDS. 85 patients with MDS and 30 healthy individuals were tested by methylation specific polymerase chain reaction (MS-PCR). The results indicated that no ZO-1 promoter methylation could be detected in healthy controls. methylation of ZO-1 gene promoter of bone marrow was found in 56.5% (48/85) MDS patients. The difference between these two kinds of subjects was statistically significant (p<0.05). The methylation status of ZO-1 gene promoter region in the subtypes of MDS was as following: RA (18/37, 48.6%), RAS (4/6, 67%), RCMD (19/30, 63%), RAEB (7/12, 58%). Every subtype of MDS patients had statistical difference from healthy people (p<0.05), but between the subtypes of MDS there were no significant statistical differences in the methylation status of ZO-1 gene, while the level of ZO-1 promoter methylation in group of RA was lower than that in other groups. It is concluded that the ZO-1 promoter region in bone marrow of MDS patient shows a hypermethylation status, which is specific for MDS. MDS is a common hematologic malignancy with clonal proliferation, it is difficult to differentiate from many other hematologic malignancies in clinical diagnosis. However, the change of ZO-1 gene methylation status is closely related to pathogenesis of MDS, therefore the ZO-1 gene as valuable diagnostic marker has important clinical significance. The ZO-1 gene may be a potential gene related to hematologic malignancies.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Methylation ; Female ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Phosphoproteins ; genetics ; Promoter Regions, Genetic ; genetics ; Young Adult ; Zonula Occludens-1 Protein

BACKGROUNDSubclinical apoplexy of pituitary functional adenoma can cause spontaneous remission of hormone hypersecretion. The typical presence of pituitary growth hormone (GH) adenoma is gigantism and/or acromegaly. We investigated the clinical characteristics of patients with spontaneous partial remission of acromegaly or gigantism due to subclinical apoplexy of GH adenoma.

METHODSSix patients with spontaneous remission of acromegaly or gigantism were enrolled. The clinical characteristics, endocrinological evaluation and imageological characteristics were retrospectively analyzed.

RESULTSIn these cases, the initial clinical presences were diabetes mellitus or hypogonadism. No abrupt headache, vomiting, visual function impairment, or conscious disturbance had ever been complained of. The base levels of GH and insulin growth factor-1 (IGF-1) were normal or higher, but nadir GH levels were all still > 1 µg/L in 75 g oral glucose tolerance test. Magnetic resonance imaging detected enlarged sella, partial empty sella and compressed pituitary. The transsphenoidal surgery was performed in 2 cases, and the other patients were conservatively managed. All the patients were in clinical remission.

CONCLUSIONSWhen the clinical presences, endocrine evaluation, biochemical examination and imageology indicate spontaneous remission of GH hypersecretion in patients with gigantism or acromegaly, the diagnosis of subclinical apoplexy of pituitary GH adenoma should be presumed. To these patients, conservative therapy may be appropriate.

Acromegaly ; diagnosis ; etiology ; Adolescent ; Adult ; Aged ; Female ; Gigantism ; diagnosis ; etiology ; Growth Hormone-Secreting Pituitary Adenoma ; complications ; Humans ; Immunohistochemistry ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pituitary Neoplasms ; complications ; Young Adult