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.

Objective To observe the seasonal changes in serum levels of interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and melatonin (MT) in Bizheng rat model, and explore the relationship between MT and the pathogenesis of rheumatoid arthritis.
Methods One hundred and sixty Sprague-Dawley rats were randomly divided into four groups in summer (n=80) and winter (n=80) respectively:normal group, collagen-induced arthritis (CIA) model group, operation group, and sham-operation group (n=20 in each group). The CIA model group was injected with collagen emulsion at the base of the tail to induce arthritis. The rats in the operation group received pineal gland resection, and 7 days after the first operation, underwent testectomy or oophorectomy. The rats in the sham-operation group were operated to ligature the sagittal sinus, without extracting the pineal gland. After the operations, the operation group and the sham-operation group both were immunized as the CIA group was.The serum levels of IL-1β, IL-6 and MT in different groups were measured by radioimmunoassay.
Results Compared with the normal group, the serum levels of IL-1βand IL-6 increased in the CIA model, operation, and sham-operation groups both in summer and in winter (IL-1βin summer, P=0.008, P<0.01, P=0.012; IL-1β in winter, P=0.019, P<0.01, P=0.027; IL-6 in summer, P=0.028, P<0.01, P=0.024;IL-6 in winter, P=0.006, P<0.01, P=0.008). In the operation group, the serum levels of IL-1βand IL-6 in winter were higher than in summer, but with no statistically significant differences (P=0.844, 0.679). Compared with the normal group, the serum level of MT significantly increased in summer and winter in both the CIA model group (P=0.002, 0.008) and the sham-operation group (P=0.003, 0.007), while significantly decreased in the operation group (P=0.023, 0.003). There was no significant difference in MT level in the operation group between summer and winter (P=0.947).
Conclusions The increase of serum levels of IL-1β and IL-6 may exacerbate the inflammatory reaction and cause a more severe condition in the rheumatoid arthritis. The concentrations of IL-1β, IL-6, and MT correspond with the change of seasons, confirming that there are connections between nature and human body.

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

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

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

Objective: This study aimed to evaluate the effects of 2 endoscopic spine surgeries on the biomechanical properties of normal and osteoporotic spines. Methods: Based on computed tomography images of a healthy adult volunteer, 6 finite element models were created. After validating the normal intact model, a concentrated force of 400 N and a moment of 7.5 Nm were exerted on the upper surface of L3 to simulate 6 physiological activities of the spine. Five types of indices were used to assess the biomechanical properties of the 6 models, range of motion (ROM), maximum displacement value, intervertebral disc stress, maximum stress value, and articular protrusion stress, and by combining them with finite element stress cloud. Results: In normal and osteoporotic spines, there was no meaningful change in ROM or disc stress in the 2 surgical models for the 6 motion states. Model N1 (osteoporotic percutaneous transforaminal endoscopic discectomy model) showed a decrease in maximum displacement value of 20.28% in right lateral bending. Model M2 (unilateral biportal endoscopic model) increased maximum displacement values of 16.88% and 17.82% during left and right lateral bending, respectively. The maximum stress value of L4–5 increased by 11.72% for model M2 during left rotation. In addition, using the same surgical approach, ROM, maximum displacement values, disc stress, and maximum stress values were more significant in the osteoporotic model than in the normal model. Conclusion In both normal and osteoporotic spines, both surgical approaches were less disruptive to the physiologic structure of the spine. Furthermore, using the same endoscopic spine surgery, normal spine biomechanical properties are superior to osteoporotic spines.

Objective: This study aimed to evaluate the effects of 2 endoscopic spine surgeries on the biomechanical properties of normal and osteoporotic spines. Methods: Based on computed tomography images of a healthy adult volunteer, 6 finite element models were created. After validating the normal intact model, a concentrated force of 400 N and a moment of 7.5 Nm were exerted on the upper surface of L3 to simulate 6 physiological activities of the spine. Five types of indices were used to assess the biomechanical properties of the 6 models, range of motion (ROM), maximum displacement value, intervertebral disc stress, maximum stress value, and articular protrusion stress, and by combining them with finite element stress cloud. Results: In normal and osteoporotic spines, there was no meaningful change in ROM or disc stress in the 2 surgical models for the 6 motion states. Model N1 (osteoporotic percutaneous transforaminal endoscopic discectomy model) showed a decrease in maximum displacement value of 20.28% in right lateral bending. Model M2 (unilateral biportal endoscopic model) increased maximum displacement values of 16.88% and 17.82% during left and right lateral bending, respectively. The maximum stress value of L4–5 increased by 11.72% for model M2 during left rotation. In addition, using the same surgical approach, ROM, maximum displacement values, disc stress, and maximum stress values were more significant in the osteoporotic model than in the normal model. Conclusion In both normal and osteoporotic spines, both surgical approaches were less disruptive to the physiologic structure of the spine. Furthermore, using the same endoscopic spine surgery, normal spine biomechanical properties are superior to osteoporotic spines.