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

AIMTo investigate the chemical constituents of the Tibetan herbal medicine Halenia elliptica by guidance of in vivo absorption and distribution of the constituents.

METHODSHPLC with a diode array detector (DAD) and HPLC-MS were used for detecting the constituents of extracts of Halenia elliptica and animal samples. Several kinds of column chromatography were used for the isolation and purification of the main in vivo absorbed and distributed compounds from the extract of Halenia elliptica.

RESULTSSix main components detected in the extracts of the animal samples were isolated from the ethanolic extract of Halenia elliptica.

CONCLUSIONAfter rats were treated with different extracts of Halenia elliplica, low polar components xanthone aglycon of Halenia elliptica were clearly observed in the extracts of liver, lipid, blood, hidney, heart and brain tissue of rats, while the polar components xanthone glycosides were detected in very small amounts in the animal samples. The xanthone glycosides can be decomposed into xanthone aglycons during the digestion, absorption and metabolism procedure. Therefore, the in vivo activity of the xanthone glycosides might be exhibited by their decomposed products. It is an accessibly valuable method to investigate chemical components of herbal medicines under the guidance by detecting in vivo absorption and distribution of chemical components of the herbal medicine extract.

Absorption ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; Female ; Gentianaceae ; chemistry ; Glycosides ; isolation & purification ; pharmacokinetics ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Tissue Distribution ; Xanthones ; isolation & purification ; pharmacokinetics

OBJECTIVETo investigate the possible effects and roles of bodyweight on the puberty onset in adolescent girls.

METHODSTotally 288 Chinese female children and adolescent girls aged 5 to 16 were followed up yearly for four consecutive years. The height, bodyweight, fat percentage, sexual characteristics, and the serum levels of leptin and insulin-like growth factor-1 (IGF-1) were studied to analyze the influential factors of puberty onset and age of menarche.

RESULTSThe serum level of leptin elevated significantly from age 13 [(9.23 +/- 1.25) microg/L] and reached peak at age 16 [(13.19 +/- 1.45) microg/L]. IGF-1 significantly correlated with the timing of puberty onset (r = 0.292, P = 0.016). BMI and fat percentage had no significant effects on the onset of puberty, but were negatively correlated with the age of menarche (r = -0.323, P = 0.037, r = -0.298, P = 0.038 respectively).

CONCLUSIONBodyweight may have effect on puberty onset in female adolescents.

Adolescent ; Adolescent Development ; Body Weight ; Child ; Child, Preschool ; Cross-Sectional Studies ; Female ; Follow-Up Studies ; Humans ; Puberty ; physiology ; Young Adult

OBJECTIVETo compare the expressions of lysyl oxidase (LOX) and matrix metalloproteinases-2 (MMP-2) in gastric cancer and pericancerous tissues, in gastric cancers with and without lymph node metastasis, and to analyze the effects of LOX and MMP-2 on tumor invasion and metastasis.

METHODSGastric cancer and pericancerous tissues were collected from 46 patients who underwent surgery. Levels of LOX and MMP-2 mRNA were detected by RT-PCR. Protein abundance of LOX and MMP-2 was examined using Western blot.

RESULTSExpressions of LOX and MMP-2 mRNA, and protein in 46 gastric cancers were significantly higher than that in 46 pericancerous tissues. In gastric cancer with lymph node metastasis, the levels of LOX and MMP-2 mRNA and protein were higher than those in gastric cancers without lymph node metastasis (P < 0.05). In the groups of gastric cancer with lymph node metastasis, expression of LOX was positively correlated with MMP-2 protein expression (P < 0.01).

CONCLUSIONSExpressions of LOX and MMP-2 in gastric cancer tissues are significantly higher than that in pericancerous tissues. The expressions of LOX and MMP-2 in gastric cancer with lymph node metastasis are higher than that in gastric cancer without lymph node metastasis. Expressions of LOX and MMP-2 are positively correlated. The results suggest that LOX and MMP-2 may promote the growth and metastasis of gastric cancer.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Biomarkers, Tumor ; metabolism ; Female ; Gastrectomy ; Humans ; Lymphatic Metastasis ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Middle Aged ; Neoplasm Invasiveness ; Protein-Lysine 6-Oxidase ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Stomach ; metabolism ; surgery ; Stomach Neoplasms ; metabolism ; pathology ; surgery

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

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