Aim To investigate the effects of berberine on epithelial-mesenchymal transition (EMT) of human liver cancer HepG2 cells induced by transforming growth factor-pi ( TGF-pl ) and its mechanism. Methods MTT assay was used to detect the proliferation activity of berberine on HepG2 cells. After 10 ng • L"1 TGF-pl was used to induce EMT model process of HepG2 cells, berberine was added to treat HepG2 cells. Colony formation, cell scratch and Transwell assays were used to detect the clonogenic, migratory and invasive capabilities of HepG2 cells. Immunofluorescence assay was used to detect the expression of EMT mesenchymal marker Vimentin. Western blot assay was used to detect the proteins expression of EMT marker (E-cadherin, N-cadherin, Snail), matrix metallopro-teinase ( MMP-2), TGF-p/Smad pathway (Smad2, p-Smad2, Smad3, p-Smad3) in HepG2 cells. Results Berberine inhibited the proliferation of HepG2 cells in a concentration-time dependent manner. Compared with TGF-pl group, berberine could significantly inhibit the abilities of colony formation, migration and invasion of HepG2 cells. Berberine could significandy inhibit the expression of E-cadherin protein up-regula-ted by TGF-pl, and N-cadherin, Vimentin;, Snail, MMP-2, p-Smad2, p-Smad2 proteins expression down-regulated by TGF-pl. Conclusions Berberine may interfere with the EMT process of HepG2 cells induced by TGF-pl by inhibiting the TGF-p/Smad 'signaling pathway to inhibit the HepG2 cell migration and invasion.

Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is an autosomal recessive disease caused by SLC25A13 gene mutations, and is characterized by delayed jaundice clearance, liver dysfunction, and elevated aminoacidemia. The confirmed diagnosis depends on gene analysis. Citrin deficiency is one of the important causes of neonatal intrahepatic cholestasis in China. Recently more and more researches about NICCD were reported. The paper summarized the epidemiology, pathogenesis, clinical characteristics, and progresses in diagnosis and treatment of NICCD.

OBJECTIVETo research the effects of Guizhi Fuling capsule on sex hormones levels in blood serum and breast issue morphology of hyperplasia of mammary glands model rats.

METHODThe unpregnancy SD rat models of hyperplasia of mammary glands were established by injecting 0.5 mg x kg(-1) benzoate estradiol. After five weeks doses,the effects of Guizhi Fuling capsule 2.0, 1.0, 0.5 g x kg(-1) and Rupixiao tablet 0.5 g x kg(-1) on the changes of papilla diameter, height and breast issue morphology of the naimal models were explored, and sex hormones levels in blood serum were measured.

RESULTGuizhi Fuling capsule can inhibitnipple swell, improve breast tissue morphology pathological profiles of the animal models, and decrease oestradiol (E2) level and increase progesterone (P) level in blood serum.

CONCLUSIONThese results suggested that Guizhi Fuling capsule could, improve mammary gland pathological profiles. Regulating sex hormone levels may be its important mechanism for treatment of hyperplasia of mammary glands.

Animals ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gonadal Steroid Hormones ; blood ; Hyperplasia ; Mammary Glands, Animal ; drug effects ; pathology ; Rats

Objective To evaluate left ventricular(LV)myocardial contraction patterns and function when pacing in different right ventricular(RV)sites and discuss echocardiogarphic method to evaluate physiologcal pacing mode.Methods This study included 26 patients with paroxysmal supraventricular tachycardia without organic heart disease.Four pacing modes including right atrium pacing(AAI),RV apex pacing(VVI-RVA),RV septal pacing(VVI-IVS)and RV outflow tract pacing(VVI-RVOT)were performed on the patients in a random order after succussful radiofrequency ablation.The parameters measured in each pacing mode included(1)LV systolic function parameters:LV twist angle(Twist),aortic systolic velocity-time integral(VTIAo)and LV global strain(Gε)；(2)LV contracting pattern:segmental peak systolic strain(Sε),the time to peak value(TPε),and the distribution of segmental Sε,TPε in each layer or wall.The relationship between Sε,TPε of each wall was analyzed.[Results]Pacing from RV sites showed lower Twist,VTIAO and Gε than AAI mode.Gε demonstrated significant difference in three RV sites pacing mode(VVI-RVOT＞VVI-IVS＞VVI-RVA,P＜0.05).Compared with the AAI mode,the distribution of segmental Sε,TPε in the each layer or wall alerted significantly in three RV sites pacing mode,especially in VV1-RVA.The distribution pattern was similar in VVI-RVOT and VVI-IVS.Furthermore,the wall Sε collated negtively with wall TPε(r =-0.51,P＜0.001).[Conclusions]Compared with AAI mode,RV pacing,especially the VVI-RVA induced the alternation of LV contraction patterns and reduction of systolic function.Longitudinal strain parameters can be used to assess the myocardial contraction patterns and function in different pacing mode.

OBJECTIVETo investigate the distribution of γ-glutamyl hydrolase gene (GGH) 452C/T genotype and allele frequency in children with acute leukemia (AL) and healthy children.

METHODSBone marrow samples from 92 children with AL and peripheral blood samples from 124 healthy children were obtained to prepare complementary DNAs (cDNAs). The cDNAs were analyzed for a GGH 452C/T polymorphism by reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis (RT-PCR-DGGE) and direct sequencing.

RESULTSThe frequencies of the AL patients with TT, CT and CC genotypes were 2.2%, 13.0% and 84.8%, and the frequencies of the control children were 1.6%, 16.9% and 81.5%, respectively. There was no significant difference in GGH genotype or T allele frequency between the two groups (P> 0.05). However, the T allele frequency in Han Chinese children was significantly different from those reported in Japanese, Mexican and African-American populations.

CONCLUSIONThe frequency of 452C/T polymorphism of GGH gene in Han Chinese children has been determined. The results suggested that an ethnic difference may exist.

Acute Disease ; Base Sequence ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genotype ; Humans ; Infant ; Infant, Newborn ; Leukemia ; enzymology ; genetics ; Male ; Polymorphism, Single Nucleotide ; gamma-Glutamyl Hydrolase ; genetics

OBJECTIVETo study the association between methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms and toxicities after high-dose methotrexate (HD-MTX) infusion in children with acute lymphocytic leukemia (ALL).

METHODSMTHFR variants in 52 children with ALL were determined by reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis and sequencing. Toxicities of children who received HD-MTX chemotherapy were evaluated according to the National Cancer Institute-Common Toxicity Criteria (NCI-CTC).

RESULTSThe children carrying MTHFR 1298AC had a higher risk of developing thrombocytopenia compared with the carriers of the 1298 AA genotype (OR=13.7, 95%CI=1.18-159.36, P=0.036). There was no significant difference in HD-MTX chemotherapy-related adverse effects between the patients with different MTHFR C677T or G1793A genotypes.

CONCLUSIONSMTHFR A1298C polymorohism may associate with the toxicity of HD-MTX chemotherapy in children with ALL.

Antimetabolites, Antineoplastic ; adverse effects ; Child ; Child, Preschool ; Female ; Genotype ; Humans ; Male ; Methotrexate ; adverse effects ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Genetic ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

This study was aimed to clone the gene coding mouse CXC chemokine receptor 4 (CXCR4), to construct the recombinant lentiviral vector carrying enhanced green fluorescence protein (EGFP) and to explore its expression in eukaryotic cells (293FT cells). The full length CXCR4 gene was cloned by RT-PCR using bone marrow cells from C57BL/6 mouse as template and inserted into PCR-Blunt vector. CXCR4 fragment was generated by digestion with restriction endonuclease and subcloned into a lentiviral vector to generate recombinant lentiviral vector LV-IRES-EGFP-CXCR4, which was co-transfected into 293FT cells together with envelope plasmid and packaging plasmid by lipofectamine 2000. Viruses were gathered and concentrated using ultracentrifuge, and then transfected into 293FT cells. Expression of EGFP was detected by fluorescent microscopy and flow cytometry (FCM). And the expression of CXCR4 protein was detected by Western blot. The results demonstrated that mouse CXCR4 gene was cloned and the lentiviral vector was successfully constructed. The lentiviral particles were correctly packaged, and the virus titers were above 10(8) TU/ml in the supernatant after concentration. Expression of EGFP was detected by fluorescent microscopy in the transfected 293FT cells, and the transfection efficacy > 95% was determined by FCM. Expression of CXCR4 protein detected by FCM and Western blot was significantly higher than that in control group. It is concluded that the CXCR4 gene along with the gene coding EGFP is successfully inserted into a lentiviral vector to construct a recombinant lentiviral vector, which can be expressed in eukaryotic cells.
Animals ; Cell Line ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lentivirus ; genetics ; Mice ; Mice, Inbred C57BL ; Plasmids ; Receptors, CXCR4 ; genetics ; Transfection

PURPOSE: The aim of the study was to evaluate the efficacy and safety of combining sorafenib with chemotherapy in patients with human epidermal growth factor receptor 2 (HER2)-negative advanced breast cancer. METHODS: MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, American Society for Clinical Oncology abstracts, and European Society for Medical Oncology abstracts were searched. Randomized clinical trials that compared the efficacy and safety of sorafenib plus chemotherapy in patients with HER2-negative advanced breast cancer with placebo plus chemotherapy were eligible. The endpoints were progression-free survival (PFS), overall survival (OS), time to progression (TTP), duration of response (DOR), overall response rate (ORR), clinical benefits, and adverse effects. The meta-analysis was performed using Review Manager 5.2.6 (The Nordic Cochrane Centre), and the fixed-effect model weighted by the Mantel-Haenszel method was used. When considerable heterogeneity was found (p<0.1), further analysis (subgroup analysis, sensitivity analysis, or random-effect model) was performed to identify the potential cause. The results are expressed as hazard ratios or risk ratios, with their corresponding 95% confidence intervals. RESULTS: The final analysis included four trials comprising 844 patients. The results revealed longer PFS and TTP, and higher ORR and clinical benefit rates in patients receiving sorafenib combined with chemotherapy compared to those receiving chemotherapy and placebo. OS and DOR were similar in the two groups. Meanwhile, the incidence of some adverse effects, including hand-foot skin reaction/hand-foot syndrome, diarrhea, rash, and hypertension, were significantly higher in the sorafenib arm. CONCLUSION: Sorafenib combined with chemotherapy may prolong PFS and TTP. This treatment was associated with manageable toxicities, but frequent dose interruptions and reductions were required.
Arm ; Breast Neoplasms* ; Breast* ; Diarrhea ; Disease-Free Survival ; Drug Therapy* ; Exanthema ; Humans ; Hypertension ; Incidence ; Medical Oncology ; Odds Ratio ; Population Characteristics ; Receptor, Epidermal Growth Factor ; Skin ; Treatment Outcome

OBJECTIVETo explore the methylation status of 5' CpG island of fragile histidine triad (FHIT) gene in plasma and the expression of FHIT protein in cancer tissue of cervical cancer patients.

METHODSMethylation-specific PCR (MS-PCR) was employed to examine methylation of FHIT gene in 151 plasma samples before treatment. The immunohistochemistry was used to the expression of FHIT protein in cancer tissues.

RESULTSCpG island methylation of FHIT was detected in 31.13% of plasma samples. The expression of FHIT protein was decreased or discarded in 59.60% of cervical cancer tissues. Among them 47.78% was included in methylation positive samples.

CONCLUSIONCpG island methylation of FHIT gene in plasma plays an important role on cervical cancer, which results in decreased expression of FHIT protein. It can be used to diagnose and evaluate the effect of treatment to cervical cancers.

Acid Anhydride Hydrolases ; blood ; genetics ; metabolism ; Adult ; Aged ; DNA Methylation ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasm Proteins ; blood ; genetics ; metabolism ; Polymerase Chain Reaction ; Uterine Cervical Neoplasms ; blood ; genetics ; metabolism

OBJECTIVETo investigate the effect of propofol at doses for different anesthesia depths on γ-aminobutyric acid (GABA) in different cerebral regions at propofol uptake equilibrium in dogs.

METHODSTwelve 12-18-month-old healthy hybrid dogs weighing 10-12 kg were randomly divided into light anesthesia group (n=6) and deep anesthesia group (n=6) with a single bolus dose of propofol (5.5 and 7.0 mg/kg, respectively) completed in 15 s followed by intravenous propofol infusion at a constant rate (55 and 70 mg·kg(-1)·h(-1), respectively). Blood samples (2 ml) were taken from the internal carotid artery and jugular vein to measure plasma propofol concentrations 50 min after the start of the infusion. The dogs were then sacrificed and tissues were taken from different brain regions and the cervical cord to measure GABA concentrations using high-pressure liquid chromatography (HPLC).

RESULTSThe plasma propofol concentrations in internal carotid artery and jugular vein were similar in both light anesthesia group (3.00 ± 0.31 and 3.10 ± 0.51 µg/ml, respectively, P>0.05) and deep anesthesia group (6.41 ± 0.05 and 6.40 ± 0.11 µg/ml, respectively, P>0.05). GABA concentrations in the brain regions were significantly higher in deep anesthesia group than in light anesthesia group (P<0.05). The dorsal thalamus and hypothalamus showed greater GABA variations [(83.83 ± 2.230%) and (85.83 ± 1.72)%] compared to other brain regions at different anesthesia depths (P<0.05).

CONCLUSIONSIn both groups, plasma propofol concentrations in the internal carotid artery and internal jugular vein reach equilibrium at 50 min of propofol infusion. The variation of GABA is associated with the anesthesia depth of propofol, and GABA variation in the dorsal thalamus and hypothalamus plays an important role in propofol anesthesia.

Anesthetics, Intravenous ; pharmacokinetics ; Animals ; Brain ; metabolism ; Dogs ; Female ; Male ; Propofol ; blood ; pharmacokinetics ; gamma-Aminobutyric Acid ; metabolism