Objective To examine the expression of hypoxia inducible factor-1 α (hypoxia-inducible factor 1α,HIF-1α),tumor cell proliferation antigen Ki67 and vascular endothelial growth factor (VEGF) in breast cancer tissues,adjacent tissues and normal breast tissues.Methods Immunohistochemistry HRP method was used to detect the expression of HIF-1α,Ki67 and VEGF in 70 cases of breast cancer tissues and their adjacent tissues and 10 cases of normal breast tissues.Difference of the positive rate was assessed viax2 test,and the correlation between the 3 were analyzed in a disorderly classified manner.Results The positive expression rate of HIF-1α,Ki67 and VEGF in 70 cases of breast cancer tissues was 78.6％ (55/70),88.6％ (62/70) and 65.7％ (46/70) respectively,significantly higher than that in adjacent tissues and normal breast tissues (P ＜ 0.01).The positive expression rate of HIF-1 α,Ki67 and VEGF was closely related with clinical stage,lymph node metastasis and histological grade (P ＜0.05).The expression intensity of HIF-1α was positively correlated with Ki67 and VEGF in breast cancer (P ＜ 0.05).Conclusions Detection of HIF-1 α,Ki67,VEGF may be an important indicator to predict the development,invasion,metastasis and prognosis of breast cancer.Tissue hypoxia may promote the proliferation of breast cancer cell and angiogenesis.A presumption is that HIF-1αmay be a new t()get of breast cancer,thus it deserves further study.

OBJECTIVETo observe the effect of Astragalus Injection (ASI) combined with rat's mesenchymal stem cells transplantation (rMSCs) for repairing spinal cord injury in rats.

METHODSOne hundred and twenty Wistar rats were randomly divided into 6 groups, named respectively by letters from A-F. they were treated respectively with none, PBS solution, ASI, rMSCs, and ASI + rMSCs, with the ASI administered via intraperitoneal injection and the rMSCs given by local injection to the spinal cord, on the 3rd day of operation. The condition of nerve function recovery was assessed on the 7th, 14th, 21st and 28th day after treatment by scoring according to the Basso-Beattie-Bresnahan (BBB) locomotor rating scale. Besides, the pathological change of the injured spinal cord was observed and expressions of glial fibrillary acidic protein (GFAP) and neurofilament-M (NF-M) in the BrdU-labeled rMSCs in the spinal cord tissue were examined by immune histochemistry.

RESULTSThe recovery of spinal cord in Group D, E and F was better than that in Group B and C, showing higher BBB scores. As compared with Group E, Group F showed a higher score of nerve function and a milder inflammatory cell infiltration with lessened tissue edema in the spinal cord and more active proliferation of gliacyte. Double-labelled immunohistochemical examination showed that the transplanted rMSCs were alive in the host's spinal cord, revealing the expressions of GFAP and NF-M from the 7th day after transplantation, which were migrating to the injured site. The amount of GFAP and NF-M positive cells in Group F was much more than that in Group E (P < 0.05)

CONCLUSIONTransplantation of rMSCs is an effective method in treatment of the spinal cord injury; ASI has the capacity for inducing rMSCs differentiated into neurons, and could synergize with rMSCs to promote the repairing of the spinal cord from injury.

Animals ; Astragalus Plant ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Mesenchymal Stem Cell Transplantation ; Nerve Regeneration ; Phytotherapy ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; therapy

Objective To investigate the relationship of Maternal-fetal interface local cytokine GM-CSF and its receptor with the occurrence of early spontaneous abortion. Methods From August 2009 to December 2011,we collected 30 villi tissue samples with artificial abortion and 30 villi tissue samples with spontaneous abortion. At the same time, we collected 30 villi tissue samples with artificial abortion and 30 decidua tissue samples with spontaneous abortion,and 30 decidua tissue samples with spontaneous abortion. The human chorionic gonadotropin ( HCG) was detected by radioimmunoassay in every group. The expressions of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR ) were detected by Immunohistochemical staining and Western Blot in the villi tissue and decidua tissue in every group. Results The concentration of HCG in the spontaneous abortion group was lower than that in the artificial abortion group ( <0.05) . The protein expressions of GM-CSF and GM-CSFR were found in villus and deciduas tissues in both groups. The protein expression levels of GM-CSFR in the villus tissues were higher in spontaneous abortion group than those in artificial abortion group ( <0.05), the protein expression of GM-CSF was upregulated, but there was no statistically significant difference between two groups. In deciduas tissues, the protein expressions of GM-CSF and GM-CSFR were upregualted in spontaneous abortion group ( <0.05) . Conclusions The suitable concentrations of GM-CSF and GM-CSFR in decidua tissue maintain the pregnancy continued. However,the higher concentrations of GM-CSF and GM-CSFR in the decidua tissue may be one of reasons of spontaneous abortion.

Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.

OBJECTIVETo understand the HBV infection rate of peripheral blood mononuclear cells (PBMCs) from fetuses of HBsAg positive mothers, associated risk factors and to explore the clinical significance of detecting HBV infected PBMCs.

METHODSSixty eight pregnant women who were delivered at the First Hospital of Xi'an Jiaotong University, China from August 1995 to February 1997, and their newborns were studied. They were divided into two groups according to their status of HBV serological markers. The study group included 50 cases who were serum HBsAg positive and 18 cases without any HBV serum markers served as control group. All these cases had no symptoms of hepatitis, high risk premature labor, premature delivery and hypertensive disorder complicating pregnancy. Age and gestational age were matched in two groups. Blood samples (5 mL) were taken from the peripheral vein of pregnant women before delivery and from newborns within 24 h after birth, before inoculation of HBV vaccine (HBVac) and injection of hepatitis B immunoglobulin (HBIG). PBMCs were isolated. The sera and PBMCs were stored at -80 degrees C. HBV-DNA in serum and PBMCs were detected with nested polymerase chain reaction (n-PCR). Two pairs of oligonucleotide primers, the outer primer pair for first PCR and inner primer pair for second PCR, designed according to region S of HBV genome were synthesized by Shanghai Cell Biology Institute of Chinese Academy of Science.

RESULTSThe detection rate of HBV-DNA in serum and PBMCs from HBsAg positive pregnant women was 60.0% (30/50) and 40.0% (20/50), respectively. The detection rate of HBV-DNA in serum and PBMCs from newborns of HBsAg positive pregnant women was 46.0% (23/50) and 30.0% (15/50), respectively. Ten newborns were HBV-DNA positive in serum only, 2 were positive in PBMCs only and 13 were positive in both serum and PBMCs. In the control group, HBV-DNA was not detected in PBMC nor in serum. The positive rate of HBV-DNA in PBMCs of newborns was significantly higher in the group of mothers who were HBV-DNA or HBeAg positive in serum (P < 0.05, P < 0.01); the positive rate was significantly higher in the group of mothers who were HBV-DNA positive in both serum and PBMC than that in the group of mothers who were serum HBV-DNA positive only (P < 0.01); and it was markedly higher in the group of mothers who were PBMC HBV-DNA positive than that in group of mothers who were HBV-DNA negative in PBMCs (P < 0.01). The positive rate of HBV-DNA in PBMCs of newborns was significantly higher in the group of newborns who were HBV-DNA positive in serum than that in the group of newborns who were HBV-DNA negative in serum (P < 0.05).

CONCLUSIONSThe positive rate of HBV-DNA in PBMCs from newborns of HBsAg positive pregnant women was 30.0% (15/30). It was related to HBV viremia level and HBV-DNA status in PBMCs of mothers and newborns. Detection of HBV-DNA in PBMCs may be an important supplementary method to determine intrauterine HBV infection, and can predict the response to HBV vaccine.

Adult ; Case-Control Studies ; DNA, Viral ; blood ; Female ; Hepatitis B Vaccines ; administration & dosage ; Hepatitis B virus ; immunology ; isolation & purification ; Humans ; Immunoglobulins ; administration & dosage ; Infant, Newborn ; blood ; Infectious Disease Transmission, Vertical ; prevention & control ; Injections, Intramuscular ; Leukocytes, Mononuclear ; virology ; Male ; Mothers ; Polymerase Chain Reaction ; Pregnancy ; blood ; Risk Factors ; Time Factors ; Treatment Outcome

The borneol was included with β-CD and prepared Fufang Danshen intestinal adhesion pellets. GC method for determination of borneol in Fufang Danshen intestinal adhesion pellets was established to study its in vitro dissolution and make a comparison with the Fufang Danshen tablet, in this way, the rationality of dosage form was evaluated. The first method of dissolution determination was used for determining the in vitro dissolution of borneol in Fufang Danshen intestinal adhesion pellets in artificial intestinal juice, and Fufang Danshen tablet in artificial gastric juice and intestinal juice, respectively. Result shows: the concentration of borneol in Fufang Danshen intestinal adhesion pellets and Fufang Danshen tablet was 0.79% and 0.80%, respectively. Its in vitro dissolution was nearly 70% within 12 h in Fufang Danshen intestinal adhesion pellets, and in Fufang Danshen tablet, the dissolution was about 60% within 20 min and more than 90% within 40 min, and in artificial gastric juice, was less than 20% within 40 min but more than 80% till 150 min. Research suggests that in comparison with Fufang Danshen tablet, in vitro dissolution of borneol in the Fufang Danshen intestinal adhesion pellets showed an obvious sustained release behavior. The borneol in Fufang Danshen intestinal adhesion pellets was included with β-CD and prepared enteric preparations. To some extent, the stimulation on stomach and intestinal mucosa can be reduced and safety can be improved.
Bornanes ; adverse effects ; chemistry ; pharmacology ; Chemistry, Pharmaceutical ; methods ; Dosage Forms ; Drugs, Chinese Herbal ; adverse effects ; chemistry ; pharmacology ; Humans ; Intestinal Mucosa ; drug effects ; metabolism ; Models, Biological ; Solubility

OBJECTIVETo investigate the expression of human annexin-V (HA-V) in relation to HBV infection in different fetal tissues.

METHODSImmunohistochemistry was employed to detect the expression and distribution of HA-V in the liver, kidney, ovary, heart, fallopian tube, spleen, and thymus gland of human fetus.

RESULTSHA-V expression was detected in different tissues including the ovary, liver, intrahepatic bile duct, heart, kidney, lymphocytic cells in the thymus gland, epithelial cells of the fallopian, and cortical and medullary cells of the spleen. HA-V was distributed mainly in the cytoplasm of the cells. The liver tissues exhibited greater gray scale for HA-V expression than in the other tissues (P<0.05) and no significant difference was observed in the other tissues than the liver (P>0.05) in image analysis with Photoshop 7.0.

CONCLUSIONHA-V is an inherent protein in fetal tissues with possible relation to HBV infection of different tissues as a HBV receptor. Greater amount of HA-V in the liver may account for the vulnerability of the liver to HBV infection.

Annexin A5 ; analysis ; Fetus ; chemistry ; virology ; Hepatitis B ; metabolism ; virology ; Hepatitis B virus ; growth & development ; Humans ; Immunohistochemistry ; Liver ; chemistry ; virology ; Tissue Distribution

OBJECTIVETo explore the association of polymorphisms in folate metabolism genes, methionine synthase reductase (MTRR) gene and 5,10-methylenetetrahydrofolate reductase (MTHFR) gene, with complex congenital abnormalities and to further investigate its association with complex congenital abnormalities derived from three germ layers.

METHODSA total of 250 cases of birth defects (with complex congenital abnormalities including congenital heart disease, neural tube defects, and craniofacial anomalies) in Shanxi Province, China were included in the study. MTRR single nucleotide polymorphism (SNP) (rs1801394) and MTHFR SNP (rs1801133) were genotyped by the SNaPshot method, and the genotyping results were compared with those of controls (n=420).

RESULTSSNPs rs1801394 and rs1801133 were associated with multiple birth defects. For the recessive model, individuals with GG genotype at rs1801394 and CC genotype at rs1801133 had a relatively low risk of developing birth defects, so the two genotypes were protective factors against birth defects. The homozygous recessive genotype at rs1801133, which served as a protective factor, was associated with ectoderm- or endoderm-derived complex congenital abnormalities, while the homozygous recessive genotype at rs1801394, which served as a protective factor, was associated with ectoderm-, mesoderm- or endoderm-derived complex congenital abnormalities.

CONCLUSIONSAmong the Chinese population in Shanxi Province, the SNPs in folate metabolism genes (MTRR and MTHFR) are associated with complex congenital abnormalities and related to ectoderm, mesoderm or endoderm development.

Asian Continental Ancestry Group ; genetics ; China ; Congenital Abnormalities ; genetics ; Ferredoxin-NADP Reductase ; genetics ; Genotype ; Germ Layers ; embryology ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVETo compare the bidirectional effect of rhubarb total anthraquinone (TA) and total tannins (TT) on rats' liver.

METHODSOne hundred rats were randomly divided into 10 groups, i.e., the blank group, the model group, the blank + high dose TA group, the blank +low dose TA group, the blank + high dose TT group, the blank + low dose TT group, the model + high dose TA group, the model + low dose TA group, the model +high dose TT group, and the model + low dose TT group, 10 in each group. The carbon tetrachloride (CCI4) was used to prepare the acute liver injury rat model. TA and TT of rhubarb (at 5.40 g crude drugs/kg and 14.69 g crude drugs/kg) were intragastrically administrated to rats in all groups except the blank group and the model group, once daily for 6 successive days.The general state of rats, biochemical indices such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), laminin (LN), hyaluronic acid (HA), transforming growth factor beta1 (TGF-beta1), as well pathological results of rat liver tissues. Finally the protection laws of TA and TT for rats' liver were analyzed using factor analysis.

RESULTSCompared with the blank control group, all biochemical indices increased in the blank group (P < 0.05, P < 0.01). HA also increased in the blank + high dose TA group; AST, ALT, and HA also increased in the blank +high dose TT group (P < 0.05). Compared with the model group, AST, ALT, ALP, HA, and TGF-beta1 significantly decreased in the model + low dose TA group, the model + high dose TA group, the model + low dose TT group (P < 0.05, P < 0.01). Serum AST, ALT, and ALP also decreased in the model + high dose TT group (P < 0.05, P < 0.01). Pathological results showed that mild swollen liver cells in the model + high dose TA group. Fatty degeneration and fragmental necrosis around the central veins occurred in the blank + high dose TA group. The pathological injury was inproved in the model +low dose TA group. Two common factors, liver fibrosis and liver cell injury, were extracted by using factor analysis. TA showed stronger improvement of the two common factors than TT.

CONCLUSIONSRhubarb TA and TT showed protective and harmful effects on rats' liver. At an equivalent dosage, TA had better liver protection than TT. High dose TT played a role in liver injury to some extent.

Animals ; Anthraquinones ; adverse effects ; pharmacology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; drug therapy ; pathology ; Dose-Response Relationship, Drug ; Female ; Liver ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Tannins ; adverse effects ; pharmacology

OBJECTIVETo make preliminary study on the chemical foundation of hematopoietic effect of si-wu-tang.

METHODSi-wu-tang was fractionated by using precipitation method with ethanol and liquid-liquid extraction. Activity was investigated on radiated mice as model of blood deficiency.

RESULTAt first si-wu-tang was separated into three fractions, fraction A, B and C. Activity study showed that fraction B and C could raise the amount of peripheral white blood cell in radiated mice. Then fraction B and C were respectively further separated into fraction B1, B2, B3 and fraction C1, C2, C3. Activity study showed that fraction C2, C3 and B3 could raise the amount of peripheral white blood cell in radiated mice. Furthermore, effect of fraction C1, C2, C3, B and paeoniflorin on hematopoietic progenitor cell in bone marrow of radiated mice were investigated. Fraction C2, C3, B and paeoniflorin could promote the proliferation of hematopoietic progenitor cell in bone marrow of radiated mice.

CONCLUSIONFraction C2, C3, B and paeoniflorin are responsible for hematopoietic activity of si-wu-tang.

Angelica sinensis ; chemistry ; Animals ; Benzoates ; isolation & purification ; pharmacology ; Bone Marrow Cells ; radiation effects ; Bridged-Ring Compounds ; isolation & purification ; pharmacology ; Cell Proliferation ; drug effects ; radiation effects ; Colony-Forming Units Assay ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Glucosides ; isolation & purification ; pharmacology ; Hematopoietic Stem Cells ; cytology ; Leukocyte Count ; Ligusticum ; chemistry ; Mice ; Mice, Inbred C57BL ; Monoterpenes ; Paeonia ; chemistry ; Rehmannia ; chemistry ; Whole-Body Irradiation