OBJECTIVETo observe the dynamic time-phase expressions of key genes of brain-gut CaM signal pathway of spleen Qi deficiency rats and the intervention effect of Sijunzi decoction.

METHODMale Wistar rats were randomly divided into the normal control group, model 14 d, 21 d, 28 d groups, and Sijunzi decoction 14 d, 21 d, 28 d groups. Except for the normal control group, the remaining groups were included into the spleen Qi deficiency model with the bitter cold breaking Qi method (ig 7.5 g · kg⁻¹ · d⁻¹ of Rheum officinale, Fructus aurantii immaturus, Magnolia officinalis preparation) and the exhaustive swimming method. On the 7th day after the modeling, the Sijunzi decoction groups were orally administered with Sijunzi decoction 20 g · kg⁻¹ · d⁻¹. The expressions of key genes CaM/CaMK II of CaM signaling pathway in hippocampus and intestine at different time points by immunohistochemical method and Western blot. At the same time, the intervention effect of Sijunzi decoction on spleen Qi deficiency rats and its mechanism were analyzed.

RESULTSpleen Qi deficiency rats showed higher intestinal CaM/CaMK II expression and lower hippocampus CaM/CaMK II expression than normal rats (P < 0.05, P < 0.01). After the treatment of Sijunzi decoction, spleen Qi deficiency rats showed reduction in intestinal CaM/CaMK II expression and increase in hippocampus CaM/CaMK II expression (P < 0.05, P < 0.01).

CONCLUSIONThe formation of spleen Qi deficiency syndrome may be related to the high expression of CaM/CaMK II in small intestine tissues and its low expression in hippocampus tissues. Sijunzi decoction may achieve the therapeutic effect in spleen Qi deficiency syndrome by reducing the CaM/CaMK II expression in intestinal tissues and increasing it in hippocampus tissues.

Animals ; Brain ; drug effects ; enzymology ; metabolism ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Calmodulin ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Intestines ; drug effects ; enzymology ; metabolism ; Male ; Qi ; Rats ; Rats, Wistar ; Spleen ; drug effects ; Splenic Diseases ; drug therapy ; enzymology ; genetics ; metabolism

Adult ; Aged ; Female ; Hepatitis B, Chronic ; complications ; therapy ; Humans ; Liver Failure ; etiology ; therapy ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged ; Treatment Outcome

Humans ; Leukemia, Myeloid, Acute ; Mutation ; Prognosis ; Remission Induction ; fms-Like Tyrosine Kinase 3

OBJECTIVETo evaluate the replication and encapsidation of HBV mutants with the truncated C gene.

METHODSThe HBV mutants with the truncated C gene were constructed by molecular cloning and PCR-based deletion in vitro. The replication and encapsidation of HBV mutants were investigated by Southern blotting, PCR and real-time fluorescence PCR respectively after transfecting the HBV mutants plasmid into HepG2 cells by using liposome.

RESULTSThe C-truncated HBV mutant vectors were constructed successfully and confirmed exactly by clone sequencing and enzymes digestion. The C-truncated HBV mutants were replication defective, however, all types of HBV DNA could be detected positive in the cytoplasm and supernatant after co-transfecting the C-truncated HBV mutants plasmid and the helper constructs into HepG2 cells. The C-truncated HBV mutants were proved to produce 3-40 folds more progeny DNA than that of the wild-type HBV by DNA quantitative assay.

CONCLUSIONThe C-truncated HBV mutants are replication-deficient and could not replicate and encapsulate in the hepatocytes when transfected solely, however, the progeny HBV-variant viruses are encapsidated more effectively to secrete into supernatant when co-transfected with the helper construct which lacks part of 5 prime-proximal HBV RNA packaging signal Epsilon.

Cell Line, Tumor ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; physiology ; Humans ; Mutation ; Plasmids ; genetics ; Transfection ; Virus Replication

BACKGROUNDTo reveal the characteristics of genotype and phenotype of HIV strains in blood and some tissues of AIDS patients.

METHODSThe virus was isolated from peripheral blood mononuclear cell (PBMC),cerebrospinal fluid (CSF)and lymph nodes of 3 AIDS patients by coculture with PBMC stimulated by PHA for 72 hours from uninfected donor. The cytopathic effect of the HIV isolates was determined in cultured MT2 cell line. The env gene sequences form proviral DNA were analyzed by GCG software.

RESULTSIn one patient,there were differences between the strains from blood and different tissues both in genotype and phenotype. The biological phenotypes of two strains from CSF were non syncytium (NSI) type, their env sequences were similar to standard CNS tropic strain (SF162).

CONCLUSIONSThe viral heterogeneity exists in different body compartments within an infected individual. The neurotropic isolate which is similar to international standard strain exists in some AIDS patients in China.

Acquired Immunodeficiency Syndrome ; virology ; Adult ; Coculture Techniques ; Female ; Genetic Heterogeneity ; Genotype ; HIV ; isolation & purification ; Humans ; Leukocytes, Mononuclear ; virology ; Lymph Nodes ; virology ; Male ; Phenotype

OBJECTIVETo explore the epidemiological characters of severe acute respiratory syndrome (SARS) in Sichuan province in order to provide evidence for prevention and control.

METHODSTo generate data on SARS in Sichuan province through descriptive and analytical studies on time, geographic distribution, population, source of infection, the way of case finding, symptom, diagnosis and treatment of the cases.

RESULTSThe peak of the epidemic last from April 16 to May 7. The number of cases in Luzhou and Guangyuan cities took up 60% of the total. Mobile population occupied 68% of the cases. Most of the patients were above the age of 20 with a sex ratio of 1.5:1 (m/f). 80% of the cases had a history of working in Guangdong province and recently returned to their hometowns. The main symptoms and signs of the SARS patients would include fever, cough and chest X-ray changes.

CONCLUSIONAll cases were imported. Fluctuation of the epidemics was mainly affected by the mobility of working population who recently returned to their hometowns. Measures concerning the prevention and control of the epidemics would mainly target on the isolation of confirmed and suspected patients who might serve as the sources of infection through setting up quarantine station, assigned hospitals and special 'fever-clinics'.

Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Severe Acute Respiratory Syndrome ; diagnosis ; epidemiology ; prevention & control

OBJECTIVETo evaluate the effects of methylation inhibitor 5-Aza-2'-Deoxycytidine (5-aza-2dc) and docetaxel (DT), alone or in combination, on the proliferation, migration, apoptosis and cell cycles of the human prostate cancer cell line PC3, and to investigate the possible mechanisms of these two drugs acting on prostate cancer in vitro.

METHODSFour groups were designed in this experiment: control, 5-aza-2dc, DT, and 5-aza-2dc + DT. The inhibitory effect of 5-aza-2dc and/or DT on the proliferation, migration and invasiveness of PC3 cells was detected by MTT, wound healing assay and cell migration assay, respectively. The apoptosis of the PC3 cells and its relationship with cell cycles were determined by Annexin V-FITC/PI assay and flow cytometry.

RESULTS5-aza-2dc and/or DT significantly increased the inhibition rate of the PC3 cells, decreased their migration distance and reduced the number of the cells that invaded the lower chamber, most significantly in the 5-aza-2dc + DT group (P < 0.05). The cell apoptosis rates of the control, 5-aza-2dc, DT and 5-aza-2dc + DT groups were (10.65 +/- 0.39)%, (16.60 +/- 0.67)%, (17.95 +/- 1.08)% and (22.98 +/- 1.18)%, respectively, with the most significant increase in the combination group (P < 0.05). Combined medication of 5-aza-2dc and DT remarkably reduced the number of cells in the G0/G1 phase, and increased that in the G2/M phase (P < 0.05).

CONCLUSION5-aza-2dc and DT, either alone or in combination, can significantly inhibit the proliferation, migration and invasiveness of PC3 cells in vitro, as well as induce their apoptosis and arrest their cell cycles in the G2/M phase, with even more significant effect when used in combination than applied alone.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Deoxycytidine ; administration & dosage ; pharmacology ; Drug Synergism ; Humans ; Male ; Taxoids ; administration & dosage ; pharmacology

OBJECTIVELong term glucocorticoid (prednisolone) treatment on human growth hormone (hGH) secretion in children and adolescents and to investigate the effectiveness and safety of the recombinant human growth hormone (rhGH) treatment.

METHODSTwelve patients (age: 10.4∓1.2 years) who were treated in Peking Union Medical College Hospital from September 1999 to November 2009 were enrolled in this study. All of them had taken prednisolone with a dose of 0.5∓2.0 mg/(kg.d) for 6~18 months. Two different hGH stimulating tests was done and their growth and development was evaluated at regular intervals. Seven patients were given rhGH with a dose of 0.1 U/(kg.d) for 6~12 months to improve their growth and development after half a year of prednisolone withdrawal when their disease conditions were improved.

RESULTSThe growth speed of these 12 children decreased significantly during prednisolone treatment compared with before prednisolone treatment (1.2∓0.3cm/year vs.3.7∓1.2 cm/year,P12 months than those with a 6~12 months course (P0.05). The growth speed of seven children who received rhGH therapy for half a year were increased from 2.2∓0.1cm/year to 7.8∓0.5cm/year (P<0.05), and then to 6.9∓0.4cm/year one year later.

CONCLUSIONSThe long-term glucocorticoid treatment can decrease the hGH secretion, and thus leads to short stature and agenesis. However, the rhGH replacement can safely and effectively improve growth and development in these children after their primary diseases are improved and glucocorticoids are withdrawn.

Adolescent ; Child ; Female ; Follow-Up Studies ; Glucocorticoids ; adverse effects ; therapeutic use ; Human Growth Hormone ; secretion ; therapeutic use ; Humans ; Male ; Recombinant Proteins ; therapeutic use ; Treatment Outcome

OBJECTIVETo study the expression of inhibitor-1 of DNA binding/differentiation-1 (Id-1) and thrombospondin-1 (TSP-1) genes in mucoepidermoid carcinoma of different malignant degree and analyze the relationship between them.

METHODSUsing immunohistochemistry (IHC) staining technique, TSP-1 and Id-1 proteins in the mucoepidermoid carcinoma of different malignant degree, including well-differentiated, moderately differentiated and poorly differentiated mucoepidermoid carcinoma, and normal salivary gland tissues were detected.

RESULTSThe positive rate of Id-1 and TSP-1 in normal salivary glands were apparently lower than that in malignant mucoepidermoid carcinoma(P = 0.000, P = 0.013). The positive rate of Id-1 in moderately and poorly differentiated mucoepidermoid carcinoma was higher than that of the well-differentiated (P = 0.001, P = 0.002). However, the positive expression of Id-1 showed no relationship between the moderately and poorly differentiated mucoepidermoid carcinoma(P > 0.05). The positive rate of TSP-1 in poorly differentiated mucoepidermoid carcinoma was less than that of the well-differentiated(P = 0.014). The positive expression of TSP-1 showed no relationship between the moderately and poorly differentiated mucoepidermoid carcinoma(P > 0.05), and the positive expression of it also showed no relationship between the moderately and well differentiated mucoepidermoid carcinoma (P > 0.05). The expression of Id-1 and TSP-1 showed negative correlation(r = -0.394, P = 0.002).

CONCLUSIONThe expression of TSP-1 may inhibit the development of the mucoepidermoid carcinoma, contrarily, the expression of Id-1 may prompt the development of the mucoepidermoid carcinoma. The expression of Id-1 and TSP-1 has negative correlation.

Aged ; Carcinoma, Mucoepidermoid ; Cell Differentiation ; DNA ; Humans ; Immunohistochemistry ; Salivary Gland Neoplasms ; Thrombospondin 1

AIMTo study the effect of antiparallel phosphorothioate triplex-forming oligonucleotide (apsTFO) matching with the shear stress response element (SSRE) of tissue factor (TF) gene promoter region on the expression of TF in endothelial cells (ECs) of rat common carotid artery stenosis.

METHODSThe model of common carotid artery middle segment stenosis was established by silica gel pipe loop ligation in SD rats. The mRNA expression and protein synthesis of TF, early growth response-1 (Egr-1) and specificity protein 1 (Sp1) were measured by in situ hybridization (ISH) and immunohistochemistry (IHC) technique. GT21-apsTFO, GT20-apsTFO, GT20-psTFO and FITC-labeled apsTFO, matching with the SSRE of TF gene promoter region, were designed, and intravenously injected into rats at 0.5 h before operation. TFO was detected 4 h after the operation, and the mRNA expression and protein synthesis of TF, Egr-1 and Sp1 were detected 6 h after the operation.

RESULTSThere were much fluorescence in vascular tissue, especially in the nuclear of ECs 4.5 h after the injection of apsTFO. The mRNA expression and protein synthesis of TF reduced by 22% - 23% with injection of GT20-apsTFO 6.5 h after stenosis (P < 0.01) and by 10% - 11% with GT21-apsTFO at the same time (P < 0.05). The inhibition by GT20-apsTFO was stronger than that of the GT21-apsTFO (P < 0.05). The expression of TF was not inhibited by the GT20-psTFO (P > 0.05). The mRNA expression and protein synthesis of Egr-1 and Sp1 did not change in the rat treated with GT20-apsTFO, GT20-psTFO and GT21-apsTFO (P > 0.05).

CONCLUSIONapsTFO could mero-inhibit the expression of TF gene but could not change the expression of Egr-1 and Sp1 protein.

Animals ; Carotid Stenosis ; genetics ; metabolism ; pathology ; Early Growth Response Protein 1 ; genetics ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; pathology ; Gene Expression ; drug effects ; Immunohistochemistry ; In Situ Hybridization ; Male ; Oligonucleotides ; chemical synthesis ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Shear Strength ; Sp1 Transcription Factor ; genetics ; metabolism ; Stress, Mechanical ; Thromboplastin ; genetics ; metabolism