Objective Androgen receptor ( AR) is extensively expressed in breast cancer and influences the proliferation of the malignant cells.Our study aimed to investigate the effect of AR on estrogen receptor (ER)-positive breast cancer. Methods ER-positive MCF-7 breast cells were exposed to various concentrations of agonist dihydrotestosterone ( DHT) or antagonist bicalutamide or left untreated .Then the proliferation and apoptosis of the cells were determined by MTT assay , cell counting , and flow cytometry , and the expressions of the proteins related to cell cycle regulation were detected by Western blot . Results The relative gray value of AR was significantly increased in the DHT group (1.055 ±0.020) but decreased in the bicalutamide group (0.705 ±0.010) as com-pared with the blank control (0.795 ±0.020) (both P<0.05).Flow cytometry showed that the early apoptosis rate of the breast cancer cells was markedly higher in the DHT group ([51.20 ±0.312]%) but lower in the bicalutamide group ([2.410 ±0.367]%) than in the blank control ([3.540 ±0.375]%) (both P<0 .01). In comparison with the control group , the expressions of the p53, p73 and p21 proteins in the MCF-7 cells were remarkably up-regulated in the DHT group but down-regulated in the bicalutamide group ( both P<0.05). Conclusion AR inhibits the proliferation of ER-posi-tive breast cancer cells , which suggests that it may be a potential ther-apeutic target for ER-positive breast cancer .

Background Trachoma is a common disease in ophthalmology,but few reports about the pathogenetic genotype are reported.Objective This study was to investigate the genotypes of Chlamydia trachomatis.Methods Conjunctival specimens were collected in 16 patients with trachoma in Beijing Tongren Eye Center from January,2003 and August,2006.Variable sequence 4 (VS4) of ompl fragment was amplified by nested PCR(n-PCR) using specific primer of Chlamydia trachomatis,and then the PCR products were sequenced.The DNA sequences were analyzed by software Clustal X and MEGA2,and the genetic characteristics were compared with the known sequences of GenBank.Results The PCR product fragment of MOMP gene of trachoma was 277 bp.Three types of Chlamydia trachomatis strains were idcntified in the 16 specimens,including B-genotype in 9 strains (56.3％),C-genotype in 4 strains(25.0％) and D-genotype in 3 strains(18.7％).High homology was seen in the gene sequences of Chlamydia trachomatis strains between the B-genotype or C-genotype strains and the same genotypes of GenBank or those from some districts of China.However,some differences were exhibited among the Dgenotype strains.Conclusions Identification of genotype of Chlamydia trachomatis has differential effect on trachoma and inclusion conjunctivitis.

OBJECTIVETo explore metabolite profiling changes in serum of rats with pi-qi deficiency syndrome (PQDS) and pi-yang deficiency syndrome (PYDS) based on liquid chromatograph-mass spectrometer (LC-MS) technique, and to explore the essence of Pi-deficiency syndrome (PDS) from small molecule metabolite level.

METHODSTotally 21 male SD rats of SPF grade were randomly divided into three groups, the normal control group, the PQDS group, and the PYDS group, 7 in each group. Rats in the PQDS group overate for 1 day and fasted for 2 days. They drank freely and then swam to be exhausted in water at 35 degrees C - 37 degrees C for 15 successive days. The PYDS model was established by the same method for PQDS rats plus drenching 20% Folium sennae water extract (2 mL/100 g), once in the morning and once in the evening for one successive week. After modeling, models were evaluated according to rat general state, changes in body weight and rectal temperature. Serum metabonomic profiles were detected using LC-MS technique. Difference in inter-group metabolite spectrograms was analyzed using orthogonal partial least squares discriminant analysis (OPLS-DA). Potential biomarkers related to syndrome types in rat serum were selected via the parameter of variable importance in the projection (VIP).

RESULTSThe weight of rats in the PQDS group and the PYDS group decreased more significantly after modeling. The difference in prepost weight was statistically significant from that of the normal control group (P < 0.01). It was more obviously lowered in the PYDS group than in the PQDS group (P < 0.05). Compared with the normal control group, the rectal temperature of rats in the PYDS group and the PQDS group decreased (P < 0.05, P < 0.01). It decresed more in the PYDS group than in the PQDS group (P < 0.05, P < 0.01). Compared with the normal control group, levels of PC(19:0)/PE(22:0), PC(17:0)/PE(20:0), capric acid, oleic acid, stearic acid, succinic acid, fumaric acid, malic acid, glucose increased; arachidonic acid, linolenic acid, lauric acid, androsterone, 4-heptanone, dihydroxyacetonephosphate (DHAP) (6:0), and uridine decreased in the PYDS group and the PQDS group. Compared with the PQDS group, levels of PC(22:1), PC (22:6), PE (18:0)/PC (15:0), retinol, and deoxycytidine increased significantly in the PYDS group; PC (18:1), PC(19 :3), PC (20:3), PC (17:0)/PE (20:0), PC (19:1)/PE (22:1), PC (19:0)/PE (22:0), PC (17:1)/PE (20: 1), PC (16:1)/PE (19:1), cholic acid, hippuric acid, furoic acid, undecanedicarboxylic acid, palmitoleic acid, hydroxy stearic acid, eicosatrienoic acid, phenylalanine, tyrosine, glutamic acid, serine, carbamoyl aspartic acid, palmitoyl carnitine, tetradecanoyl carnitine, acetylcarnitine, and linoleylcarnitine decreased more significantly in the PYDS group.

CONCLUSIONSComparative contents of various serum metabolites changed significantly in PQDS and PYQS model groups. Some potential small molecular biomarkers related to PDS were preliminarily identified. These results might provide some data reference for exploring scientific connotation and pathological mechanisms of PDS.

Animals ; Biomarkers ; blood ; Chromatography, Liquid ; Discriminant Analysis ; Disease Models, Animal ; Drugs, Chinese Herbal ; Least-Squares Analysis ; Male ; Mass Spectrometry ; Metabolome ; Metabolomics ; Qi ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; blood

OBJECTIVETo explore efficacy enhancing and detoxification roles of Jiedu Quyu Zishen Recipe (JQZR) in treating systemic lupus erythematosus (SLE) by studying its effect on Toll like receptor 9 (TLR9) signal pathway of murine macrophage cells after JQZR stimulated CpG oligodeoxynucletide (CpG ODN).

METHODSMurine macrophage cells in vitro cultured were randomly divided into 4 groups, i.e., the blank serum group, the CpG ODN stimulus group, the CpG ODN + dexamethasone group, the CpG ODN + medicated serum group. Murine macrophage cells were collected after 24-h intervention. The expression of TLR9, myeloid differentiation factor 88 (MyD88), NF-KB, IFN-α mRNA were analyzed by RT-PCR. The expression of TLR9 and NF-κB protein were analyzed by Western blot. Changes of the NF-KB transcriptional activity were assayed by Dual-Luciferase reporter assay system.

RESULTSmRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were enhanced, showing statistical difference when compared with those of the blank serum group (P <0. 05, P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of MyD88, NF-κB, and IFN-α, the protein expression of NF-κB and the NF-κB transcriptional activities decreased in the CpG ODN + dexamethasone group with statistical difference (P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were decreased in CpG ODN+ medicated serum group with statistical difference (P <0. 01).

CONCLUSIONEfficacy enhancing and detoxification roles of JQZR in treatment of SLE might be realized through regulating TLR9 signal pathways.

Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Macrophages ; metabolism ; Mice ; Myeloid Differentiation Factor 88 ; NF-kappa B ; RNA, Messenger ; Signal Transduction ; Toll-Like Receptor 9 ; metabolism

OBJECTIVETo explore the intervention of Huayu Qutan Recipe (HQR) on liver SREBP-2 signal pathway of hyperlipidemia rats of Pi deficiency syndrome (PDS).

METHODSTotally 100 SPF grade SD rats were randomly divided into the blank control group, the hyperlipidemia group, the hyperlipidemia treatment group, the PDS hyperlipidemia group, and the PDS hyperlipidemia treatment group, 20 in each group. Common granular forage was fed to rats in the blank control group. High fat forage was fed to rats in the hyperlipidemia group and the hyperlipidemia treatment group. Rats in the PDS hyperlipidemia group and the PDS hyperlipidemia treatment group were treated with excessive labor and improper diet for modeling. They were administered refined lard by gastrogavage (3 mL each time, twice per day) and fed with high fat forage on the odd days, and fed with wild cabbage freely on even days. The modeling lasted for 30 days. Rats in the hyperlipidemia treatment group and PDS hyperlipidemia treatment group were administered with Huayu Qutan Recipe (20 mL/kg) by gastrogavage, once a day, for 30 successive days. Levels of serum cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), and serum amylase (AMY) were detected by automatic biochemical analyzer. D-xylose excretion rate was determined using phloroglucinol method. Morphological changes of liver and the lipid deposition in liver were observed using HE stain and oil red O stain respectively, mRNA and protein expression levels of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR), cholesterol 7α-hydroxylase 1 (CYP7A1), LDL-R, and sterol regulatory element binding protein-2 (SREBP-2) were detected using real time RT-PCR and Western blotting.

RESULTSCompared with the blank control group, serum levels of TC (1.84 ± 0.19 mmol/L, 2.23 ± 0.43 mmol/L) and LDL-C (0.99 ± 0.24 mmol/L, 1.13 ± 0.56 mmol/L) were higher in the hyperlipidemia group and the PDS hyperlipidemia group, serum levels of HDL-C (0.41 ± 0.66 mmol/L, 0.41 ± 0.11 mmol/L) and AMY activities (351 ± 45 mmol/L, 153 ± 30 mmol/L) were lower, and urinary D-xylose excretion rates were lower (26.9 ± 2.1 ng/mL, 15.0 ± 1.7 ng/mL) (all P < 0.05). Lipid deposition occurred in liver cells. Much fat vacuoles occurred in the cytoplasm. Expression levels of HMGCR, CYP7A1, LDL-R, and SREBP-2 mRNA and proteins in liver significantly decreased (P < 0.01). Compared with the hyperlipidemia group, serum levels of TC and LDL-C significantly increased (P < 0. 05), AMY activities and urinary D-xylose excre- tion rates significantly decreased in the PDS hyperlipidemia group (P < 0.01). A large amount of lipid deposition occurred in liver. The atrophy of liver cells was obviously seen. Expression levels of CYP7A1, LDL-R, and SREBP-2 mRNA and proteins in liver were significantly lower (P < 0.01, P < 0.05). Serum levels of TC and LDL-C significantly decreased (P < 0.05), AMY activities and urinary D-xylose excretion rates significantly increased in the hyperlipidemia treatment group (P < 0.01). Expression levels of CYP7A1, LDL-R, and SREBP-2 mRNA and proteins in liver were significantly increased (P < 0.01, P < 0.05). Compared with the PDS hyperlipidemia group, serum level of TC significantly decreased (P < 0.05), HDL-C levels, AMY activities and urinary D-xylose excretion rates significantly increased in the PDS hyperlipidemia treatment group (P < 0.01),expression levels of CYP7A1, LDL-R, and SREBP-2 mRNA and proteins in liver were significantly increased (P < 0.01). Similar changes occurred in the two treatment groups.

CONCLUSIONSPi deficiency exacerbates abnormal serum TC level and the lipid deposition in liver. These might be related to regulating expression levels of LDL-R, HMGCR, and CYP7A1 genes in the SREBP-2 signal pathway. HQR could regulate this pathway to intervene abnormal metabolism of TC.

Animals ; Cholesterol, HDL ; Cholesterol, LDL ; Drugs, Chinese Herbal ; therapeutic use ; Hyperlipidemias ; drug therapy ; Liver ; Male ; Medicine, Chinese Traditional ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Sterol Regulatory Element Binding Protein 2 ; metabolism ; Triglycerides

The aqueous phase oxidation of gaseous elemental mercury (Hg(0)) by potassium persulfate (KPS) catalyzed by Ag(+) was investigated using a glass bubble column reactor. Concentration of gaseous mercury and potassium persulfate were measured by cold vapor atom absorption (CVAA) and ion chromatograph (IC), respectively. The effects of pH value, concentration of potassium persulfate and silver nitrate (SN), temperature, Hg(0) concentration in the reactor inlet and tertiary butanol (TBA), free radical scavenger, on the removal efficiency of Hg(0) were studied. The results showed that the removal efficiency of Hg(0) increased with increasing concentration of potassium persulfate and silver nitrate, while temperature and TBA were negatively effective. Furthermore, the removal efficiency of Hg(0) was much better in neutral solution than in both acidic and alkaline solution. But the influence of pH was almost eliminated by adding AgNO(3). High Hg(0) concentration has positive effect. The possible reaction mechanism of gaseous mercury was also discussed.

OBJECTIVETo investigate the therapeutic effect, long term survival and side effect on NSCLC patients treated with nadaplatin combined with paclitaxol and cisplatin combined with paclitaxol.

METHODSNSCLC patients with stage IIIB or IV were randomized into two groups in this prospective clinical study. TN group: nadaplatin 30 mg/m2 dl-3, paclitaxol 175 mg/m2 dl, repeated every 4 weeks. TP group: DDP 30 mg/m2 dl-3, paclitaxol 175 mg/m2 dl, repeated every 4 weeks.

RESULTSSixty patients were enrolled and 57 were evaluable with 30 in TN group and 27 in TP group. The overall response rate were 43.3% vs. 48.1% (P = 0.716), and the disease control rate were 86.7% vs. 88.8% in TN and TP group (P = 0.799), respectively. The median survival time was 14.3 vs. 13.0 months, and the 1- and 2-year survival rate was 62.5% vs. 59.1%, 0% vs. 5.8% in TN and TP group (P = 0.839), respectively. The rates of neutropenia and thrombocytopenia were similar in TN and TP groups whereas more patients in TP group than in TN group suffered from anemia (38.5% vs. 17.5%, P = 0.001), nausea and vomiting (82.6% vs. 35.6%, P = 0.000), fatigue (35.9% vs. 14.1%, P = 0.000) and peripheral neurotoxicity (50.0% vs. 21.9%, calculated by case, P = 0.023).

CONCLUSIONNadaplatin combined with paclitaxol is an effective treatment regimen for NSCLC patients. When compared with similar regimen with cisplatin, the response rate and survival were similar; however, nadaplatin regimen shows some superiority as regards some treatment side effect.

Adult ; Aged ; Anemia ; chemically induced ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cisplatin ; administration & dosage ; adverse effects ; Female ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Neutropenia ; chemically induced ; Organoplatinum Compounds ; administration & dosage ; adverse effects ; Paclitaxel ; administration & dosage ; adverse effects ; Prospective Studies ; Remission Induction ; Survival Analysis ; Thrombocytopenia ; chemically induced ; Treatment Outcome

OBJECTIVETo investigate the mechanism of clinical haemorrhage in an inherited coagulation factor VII (FVII) deficiency and tissue factor abnormality pedigree.

METHODSAll exons, exon-intron boundaries and the 3', 5' untranslated sequences of FVII and tissue factor (TF) genes were amplified by PCR and sequenced directly. Any mutation identified by direct sequencing was confirmed by reverse sequencing. FVII cDNA of the proband was synthesized with random primers and amplified by nest PCR.

RESULTS55C-->T heterozygous mutation located in promoter of FVII gene was identified in the proband. The heterozygous mutation was derived from his mother. Tracing the other pedigree members found that his sister had the same heterozygous mutation and the others had wild-type FVII genes. A 9363 C-->T (Arg131Trp) heterozygous polymorphism in TF gene, which was 2.63% frequency of T allele polymorphism, was found in all of the pedigree members.

CONCLUSIONIt was the first report that the -55C-->T heterozygous mutation in FVII gene and the Arg131Trp heterozygous polymorphism in TF gene explained the clinical symptom of the proband.

Adult ; DNA Mutational Analysis ; Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Heterozygote ; Humans ; Male ; Pedigree ; Polymorphism, Genetic ; Thromboplastin ; genetics

OBJECTIVETo investigate the function of abnormal fibrinogen in two inherited dysfibrinogenemia pedigrees.

METHODSRoutine coagulation tests were conducted in the probands and related family members. The antigen and activity levels of fibrinogen were detected by immunoturbidimetry assay and clauss assay, respectively. All the exons and exon-intron boundaries of the three fibrinogen genes and antithrombin gene（AT3）were analyzed by PCR amplification and direct sequencing. Routine thrombelastography (TEG) test and functional fibrinogen TEG test were both used to make a comprehensive evaluation of coagulation status and functional fibrinogen level in patients. The molecular weights of the three peptides from fibrinogen were measured by Western blot. The function of abnormal fibrinogen was assessed by fibrinogen dynamic polymerization and fibrinolysis velocity.

RESULTSThe coagulation routine tests were normal in two probands except for prolonged thrombin time (TT) and reptilase time (RT), as well as reduced activity levels of 0.5 g/L and 0.6 g/L fibrinogen, respectively. The antigen levels of fibrinogen were 2.32 g/L and 2.66 g/L in two probands, which were in the normal reference range. The genotype analysis showed that Arg275His in fibrinogen γ chain (γ Arg275His) existed in both probands and patients in these two pedigrees. Meanwhile, proband B's grandfather and aunt also carried heterozygote g.5876T>C (Ser116Pro) mutation in AT3. The results of routine TEG test demonstrated that the α values of proband B and his father were close to and lower than the lower limit of reference range, respectively, while the MA values were normal in both of them. However, functional fibrinogen TEG test revealed obviously reduced MA value. All the probands and patients demonstrated prolonged lag-off time and reduced peak value in fibrinogen dynamic polymerization tests. Meanwhile, most of fibrin formed from the patients' plasma could not be dissolved completely by plasminogen (PLG) and urokinase-typeplasminogenactivator (u-PA) at a certain time.

CONCLUSIONWe first reported cases of inherited dysgibrinogenemia associated with inherited AT deficiency. γArg275His mutation caused the abnormal fibrinogen in terms of fibrin mono polymerization and possibly in fibrinolysis. Combined use of routine TEG test and functional fibrinogen TEG test with comprehensive analyses of the parameters in both tests could better evaluate the level of functional fibrinogen and predict the risk of hemorrhage and thrombosis in patients with inherited dysfibrinogenemia.

Adult ; Afibrinogenemia ; genetics ; Child ; Female ; Fibrinogen ; genetics ; Fibrinogens, Abnormal ; genetics ; physiology ; Genotype ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

OBJECTIVETo analyze the phenotype and genotype of two Chinese pedigrees with von Willebrand diseases, and to investigate the molecular pathogenesis.

METHODSBleeding time (BT), activated partial thromboplastin time (APTT), ristocetin-induced platelet aggregation (RIPA), von Willebrand factor-ristocetin cofactor (vWF:Rco), von Willebrand factor antigen (vWF:Ag), von Willebrand factor activity (vWF:A), von Willebrand factor collagen binding assay (vWF:CB) and multimer analysis were used for phenotype diagnosis. DNA was extracted. All of the 52 exons and exon-intron bounda ries of the VWF gene were amplified with polymerase chain reaction(PCR) and analyzed by direct sequencing.

RESULTSAPTT and BT were prolonged. Plasma RIPA, vWF:Rco, vWF:Ag, vWF:A and vWF:CB was significantly decreased. No VWF multimer can be found by plasma VWF multimer analysis. Homozygous insertional mutation g.82888_82889insCATG in exon 17 was found in proband A. Compound heterozygous mutations g.94865 G to A (Trp856stop) in exon 20 and g.110698_110699delinsG in exon 28 were found in proband B.

CONCLUSIONHomozygous insertional mutation g.82888_82889insCATG and compound heterozygous mutations g.94865G to A(Trp856X) and g.110698_110699delinsG probably have respectively induced type 3 von Willebrand diseases in the two probands.

Adolescent ; Female ; Genotype ; Humans ; Male ; Mutation ; Pedigree ; Phenotype ; von Willebrand Disease, Type 3 ; genetics ; von Willebrand Factor ; genetics