AIM: To study p42/p44 mitogen - activated protein kinases ( MAPK ) signal transduction pathway effect on vascular endothelial growth factor ( VEGF ) expression induced by elevated glucose concentration in cultured human retinal pigment epithelium ( hRPE) .
METHODS:hRPE cells were cultured and divided into four groups:normal glucose group (NG) (5. 6mmol/L), high glucose group ( HG1:15mmol/L D-glucose, HG2:20mmol/L D - glucose, HG3:30mmol/L D - glucose ), PD98059 group: hRPE cells were treated by an efficient and selective inhibitor PD98059 (20μmol/L) of p42/p44MAPK signal transduction pathway and solvent dimethyl sulfoxide group ( DMSO group) . The expression of VEGF and pigment epithelium derived factor ( PEDF ) mRNA was detected by RT-PCR. VEGF protein expression in cultured hRPE supernatants was detected by enzyme-linked immumosorbent assay ( ELISA) .
RUSULTS: VEGF mRNA and protein expression induced by elevated glucose concentration increased significantly. VEGF mRNA and protein expression were restrained in PD98059 group. Ratio of ( VEGF/β-actine)/( PEDF/β - actine ) in PD98059 group decreased significantly compare with that in high glucose group.
CONCLUSION: p42/p44MAPK signal transduction pathway might play a part in VEGF expression induced by elevated glucose concentration in cultured hRPE cells.

Aged ; Diagnosis, Differential ; Fibronectins ; metabolism ; Glomerulonephritis, Membranoproliferative ; pathology ; Humans ; Immunohistochemistry ; Kidney Diseases ; metabolism ; pathology ; Kidney Glomerulus ; metabolism ; pathology ; ultrastructure ; Male ; Microscopy, Electron

To investigate the effect of huangqin tang on expression of cytokines and NF-κB p65 in rats with ulcerative colitis (UC), and to probe into its underlying mechanisms of action. The mode of UC rats with cell immunoreactivity was made using compound method (trinitrobenzene sulfonic acid and ethanol). Rats were randomly divided into control group, model group, SASP group and high dose, middle dose and low dose of huangqin tang group. The food intake, body weight and microscopic damage of rats in each group were evaluated after being treated for five days. The blood and colon tissue were also collected. Production of NO was detected by Griess assay, the expression levels of IL-6, TNF-α, PGE2 were detected by ELISA. ICH method was undertaken to determine the expression of NF-κB p65 protein in colon tissue. The food intake and body weight of model group rats were lower than that of control group. The expression levels of NO, IL-6, TNF-α, PGE2 in serum and NF-κB p65 protein of colon tissue in model group were higher than that of control group. The above indexes were ameliorated in high and middle dose of huangqin tang groups. But there was no significant difference with SASP group. NF-κB p65 may be involved in the pathogenesis of UC, and huangqin tang can inhibit the relative activity of NF-κB p65, and decrease the expression levels of NO, IL-6, TNF-α and PGE2.
Animals ; Colitis, Ulcerative ; metabolism ; Dinoprostone ; blood ; Drugs, Chinese Herbal ; pharmacology ; Interleukin-6 ; blood ; Nitric Oxide ; blood ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo study the prevalence of SEN virus (SENV) infection in CHB patients in five cities of China.

METHODSA nest-polymerase chain reaction (nPCR) was used for detection of SENV-D and SENV-H in sera of 595 CHB patients from 5 cities of China and 96 normal individuals from Beijing. A total of 7 SENV strains were analyzed by direct sequencing.

RESULTSThe prevalence rates of SENV in CHB patients and normal individuals were 61.3% and 62.5%, respectively (chi(2) = 0.047, P = 0.829). The prevalence rates of CHB patients between 5 cities were different. Nucleotide sequence analysis showed that the homology between 4 SENV-D strains was 91% - 98% and 95% - 98% between 3 SENV-H strains isolated from 5 cities in China.

CONCLUSIONSENV-D/H were prevalent in CHB patients of China and their prevalence rates were similar to that in normal individuals.

China ; epidemiology ; Circoviridae ; isolation & purification ; Circoviridae Infections ; complications ; epidemiology ; virology ; DNA Virus Infections ; complications ; epidemiology ; virology ; DNA Viruses ; isolation & purification ; DNA, Viral ; analysis ; Hepatitis B, Chronic ; complications ; virology ; Humans ; Phylogeny ; Prevalence

OBJECTIVETo investigate excretion of severe acute respiratory syndrome coronavirus RNA (SARS-CoV) in stool of SARS patients.

METHODSSARS-CoV RNA was detected in stool specimens with fluorescent quantitative polymerase chain reactions (FQ-PCR) in 101 SARS patients on the 10 to 55 days after onset, 27 non-SARS patients and 400 individuals with health check-up.

RESULTSSARS-CoV RNA was positive in stool specimens by FQ-PCR in 58 of 101 SARS patients (57.4%), and all negative in 27 non-SARS patients and 400 healthy individuals. Positive rate of SARS-CoV RNA was 100% (8/8), 67.7% (21/31), 47.4% (27/57) and 40.0% (2/5) on the 10 - 19, 20 - 29, 30 - 39 and 40 - 55 days after onset of fever, respectively, with values of logarithm of SARS-CoV RNA load of 6.06 +/- 2.05, 4.51 +/- 1.23, 3.82 +/- 1.44 and 3.57 +/- 1.25, respectively.

CONCLUSIONPositive rate and load of SARS-CoV RNA in stool of SARS patients was the highest at their acute phase, and decreased with the extension of its course.

China ; Feces ; virology ; Humans ; Polymerase Chain Reaction ; RNA, Viral ; genetics ; metabolism ; SARS Virus ; genetics ; isolation & purification ; Severe Acute Respiratory Syndrome ; virology

Objective To determine the curative effect of dilation for achalasia with temporary cardia stent in different diameters based on a long-term follow-up.Methods The study cohort was comprised of 135 patients of achalasia.Among them differentiated by stent diameters as followings:30 patients were treated under fluoroscopy with dilation of temporary cardia stent in 20 mm diameter(group A), 30 patients with dilation of temporary eardia stent in 25 mm diameter(group B),and 75 patients with dilation of temporary cardia stent in 30 mm diameter(group C).135 cardia stents were temporarily placed in the 135 patients and withdrawn after 3 -5 days via gastroscopy.All the stents were inserted and withdrawn successfully.The follow-up in all groups lasted 6-128 months.Results Six(20.0%)out of 30 patients,6(20.0%)out of 30 patients,5(22.7%)out of 22 patieuts,6(37.5%)out of 16 patients,5 out of 9 patients,3 out of 3 patients in group A exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively. Four(13.3%)out of 30 patients,4(13.3%)out of 30 patients,3(13.0%)out of 23 patients,4(22.2%)out of 18 patients,5(45.5%)out of 11 patients,and 3 out of 4 patients in group B exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively.No(0.0%)out of 75 patients, 1(1.5%)out of 66 patients,4(8.3%)out of 48 patients,6(18.2%)out of 33 patients,6(33.3%)out of 18 patients,2 out of 5 patients in group C exhibited dysphagia relapse during 6 months,1 year,3 years,5 years,8 years,and 10 years follow-up,respectively.Conclusion Dilation with temporary cardia metal stent in 30 mm diameter is the best dilation for achalasia in long-term follow-up.

OBJECTIVETo establish a simple, sensitive, specific and less-costly method for detecting genotypes of TT virus (TTV).

METHODSTTV DNA was tested by nested polymerase chain reaction (nPCR) in sera from 180 patients with different types of viral hepatitis and 96 normal individuals in Beijing. TTV genotypes were determined in 40 sera collected from TTV DNA positive patients by heteroduplex mobility assay (HMA) and through sequencing.

RESULTSThe positive rates of TTV DNA in viral hepatitis patients and normal individuals were 22.2% (40/180) and 19.8% (19/96), respectively (chi(2) = 0.220, P = 0.639). TTV DNA positive rates of patients with hepatitis A, B, C, E and non-A to E were 20.0% (6/30), 16.7% (5/30), 23.3% (7/30), 36.7% (11/30) and 18.3% (11/60), respectively. Of 40 TTV DNA positive patients, 20 (50.0%) were TTV G1, 7 (17.5%) TTV G2, 10 (25.0%) coinfected with different genotypes of TTV, and 3 untyped by HMA. Twenty G1 and 7 G2 detected by HMA were confirmed by sequence analysis. Of 10 patients coinfected with different genotypes of TTV, 5 were G1 and G2, 2 G1 and G3, 1 G1 and G4, 1 G1 and G3, and 1 with G1, G2 and G3 coinfections.

CONCLUSIONHMA was recognized as simple, sensitive, specific and less-costly, thus could be used for genotyping of TTV.

DNA, Viral ; analysis ; Genotype ; Hepatitis, Viral, Human ; virology ; Heteroduplex Analysis ; methods ; Humans ; Phylogeny ; Torque teno virus ; classification ; genetics

OBJECTIVETo study on the dynamics of peripheral blood lymphocytes and their subpopulations in patients with severe acute respiratory syndrome.

METHODSUsing flow cytometry, the absolute numbers of peripheral blood lymphocytes and their subpopulations in 240 SARS patients (696 specimens) and 51 individuals as controls, were counted and compared.

RESULTSThe absolute numbers of peripheral blood lymphocytes and their subpopulations (CD45, CD3, CD4, CD8) were 1298 +/- 785, 897 +/- 606, 510 +/- 372, 362 +/- 263/mm(3), respectively, significantly lower in SARS patients as compared to the normal controls (2024 +/- 423, 1391 +/- 289, 795 +/- 129, 551 +/- 183/mm(3)). Of SARS patients, severe group (1095 +/- 740, 740 +/- 562, 419 +/- 346, 304 +/- 244/mm(3)) had lower counts than that of mild group (1404 +/- 788, 991 +/- 612, 564 +/- 378, 396 +/- 267/mm(3)), and in group with deaths (587 +/- 493, 369 +/- 371, 204 +/- 191, 150 +/- 130/mm(3)) was lower than that of recovery group (1355 +/- 776, 948 +/- 603, 539 +/- 375, 382 +/- 263/mm(3)). There were significant differences (P < 0.01) for CD45, CD3, CD4, CD8, but with no significant difference (P > 0.05) for CD4/CD8 ratio between severe and mild, recovery and death groups. The lymphocytes and their subpopulations (CD45, CD3, CD4, CD8) declined in the 1st week and to the lowest level (977 +/- 579, 641 +/- 466, 360 +/- 275, 270 +/- 216/mm(3)) in the 2nd week. Then the lymphocytes and their subpopulations gradually increased during the recovery of the disease.

CONCLUSIONThe absolute numbers of peripheral blood lymphocytes and their subpopulations in SARS patients might be used as one of the methods for diagnosis on the severity and prognosis of the disease.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; blood ; Female ; Flow Cytometry ; Humans ; Lymphocytes ; classification ; immunology ; metabolism ; Male ; Middle Aged ; Severe Acute Respiratory Syndrome ; blood ; Time Factors

DNA, Viral ; analysis ; Genotype ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Polymerase Chain Reaction

Cell Line, Tumor