Hepatic fibrosis models were induced by CCl4 in rats. To explore vascular endothelial growth factor (VEGF), transforming growth factor-beta1 (TGFβ1) mRNA expression and bcl-2, Bax protein expression levels of intervention and explore the mechanism of the Aralia chinesis anti-hepatic fibrosis. Sixty male Sprague-Dawlley (SD) rats were randomly divided into six groups: nomal group, model group, high-dose (10 mL x kg(-1)), medium-dose (7.5 mL x kg(-1)), low-dose (5.0 mL x kg(-1)) of A. chinesis treated group and colchicine treated group. The change of liver histopathology was observed by HE and Masson staining. The mRNA of VEGF, TGF-β1 were detected by RT-PCR. The protein of Bcl-2 and Bax were detected by Western blot. In the model group liver cell obvious degeneration, necrosis, a large number of collagen fibers of the cable hyperplasia, part visible pseudolobule formation. A. chinesis large, medium, low-dose group and colchicine group liver cell degeneration and necrosis reduced A. chinesis small, medium, and high-dose group was gradually reduced trend and A. chinesis large, middle dose group degree of reduction is particularly significant. Compared with model group, A. chinesis of large, medium and small dose group and colchicine group VEGF mRNA expression, A. chinesis of large, medium-dose group TGF-β1 mRNA expression reduce (P < 0.05); compared with colchicine group, A. chinesis of large, middle dose group of VEGF mRNA expression decreased (P < 0.05); A. chinesis of large, middle dose group of TGF-β1 mRNA expression decreased (P < 0.01), and compared with colchicine group, large dose group of of TGF-β1 mRNA expression decreased (P < 0.05). Compared with model group, A. chinesis of large, medium and small dose group and colchicine group Bcl-2 protein expression reduce (all is P < 0.05). But A. chinesis of large, medium and small dose group and colchicine group of Bax protein expression were increased (P < 0.05). A. chinesis regulation of VEGF, TGF-β1 may prevent the activation of hepatic stellate cells, liver tissue by up regulating the anti-apoptotic protein Bax and down pro-apoptotic protein Bcl-2 expression, thereby to improve the degree of liver fibrosis.
Animals ; Apoptosis ; drug effects ; Aralia ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Liver Cirrhosis ; drug therapy ; genetics ; metabolism ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo study the conditions and parameters of purifying 2,3,5,4'-tetrahydroxy-stilbene-2-O-beta-D-glycoside from Polygonum multiflori.

METHODAbsorption capacity of four resins for 2,3,5,4'-tetrahydroxy-stilbene-2-O-beta-D-glycoside was compared. With the adsorption ability as indexes, the process of absorbing and purifying 2,3,5,4'-tetrahydroxy-stilbene-2-O-beta-D-glycoside from P. multiflori with S-8 macroporous resin absorbent was selected by orthogonal design.

RESULTThe S-8 resin was the best of the four resins. The optimum process condition was 50% ethanol as eluting solvent, the flow rate at 1.5 mL x min(-1), pH at 7-8, and the solution concentration at 0.2 g x mL(-1). The absorption capacity by this process was 36.89 mg x g(-1).

CONCLUSIONThe process is simple and convenient and the regeneration of resin is easy, so this method of purification is advisable.

Absorption ; Glycosides ; chemistry ; isolation & purification ; Plant Extracts ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Resins, Synthetic ; chemistry ; Stilbenes ; chemistry ; isolation & purification

To identify the active constituents in vitro and blood-absorbed ingredients in vivo from Yin Chen Hao decoction provides scientific evidence for probing its prevention and treatment mechanism on acute liver injury. An ultrahigh performance liquid chromatography quadrupole-time of flight-mass spectrometry (UPLC-QTOF/MS) method was applied for analysis of Yin Chen Hao decoction and the serum samples of mice with con-A induced acute liver injury after preventive oral administration for 14 days (the use of all laboratory animals in this study was approved by the Ethics Committee of the Naval Medical University, 19YF1459400). A total of 90 chemical constituents were identified from Yin Chen Hao decoction, mainly were flavonoids, terpenoids, tannins, quinones. 5 prototype compounds were identified in the serum, including chrysophanol, deoxyrhapontin-8-O-gallate, mussaenosidic acid, herniarin, emodin. The established UPLC-QTOF/MS method could efficiently and sensitively identify the constituents in vitro and blood-absorbed ingredients of Yin Chen Hao decoction, primarily clarify the material basis of its hepatoprotective effect, and provided a scientific basis for the quality marker selection and the pharmacodynamic material basis research on the decoction.

OBJECTIVETo study the effects of indoor air pollution and individual susceptible factors on prevalence of children's asthma and asthma-related symptoms in Shenyang city.

METHODSOn April, 2007, 8733 Han children who were under age of 12 and lived for more than 2 years in Shenyang city, were selected from five administrative areas (one primary school and two kindergartens for each area) through cluster random sampling method. Information on children's general condition, asthma and related symptoms (including stridor, stridor symptoms, persistent cough, persistent phlegm), indoor air pollution, and susceptibility history were obtained by a standard questionnaire from the American Thoracic Society. The effects of indoor air pollution on asthma and asthma-related symptoms was analyzed through χ(2) test. Logistic regression was used to research the effects of risk factors on the prevalence of asthma and asthma-related symptoms of both susceptible and non-susceptible children.

RESULTSAmong the 8733 subjects, 4420 (50.6%) were boy and 4313 (49.4%) were girl, with the age of (8.08 ± 2.88) years old. The prevalence of asthma, current asthma, cough, persistent phlegm, stridor and stridor symptom were 6.4% (559 cases), 2.5% (215 cases), 9.6% (836 cases), 4.4% (386 cases), 17.5% (1524 cases) and 2.6% (229 cases) respectively. The prevalence of asthma the boys and girls were among 7.1% (313 cases) and 5.7% (246 cases) (χ(2) = 6.916, P < 0.05); and stridor symptom for them were 19.2% (850 cases), 15.6% (674 cases) (χ(2) = 19.678, P < 0.05), respectively. Passive smoking before two years old, house decoration and pet were related to asthma of children, and there was significant difference between the two groups. The prevalence of asthma of exposed children were 7.7% (312 cases), 9.5% (159 cases), 8.0% (270 cases), 9.0% (114 cases), respectively. Compared with the non-exposed children who had asthma, the prevalence of asthma were 5.7% (400 cases), 5.4% (289 cases), 6.0% (445 cases), the value of χ(2) were 33.646, 23.944 and 16.527 respectively (all P values < 0.05). Children who had family history of asthma, family history of allergy and allergy history were also related with asthma, the prevalence of asthma were 17.3% (106 cases), 13.1% (85 cases), 22.0% (147 cases), compared with the non-exposed children who had asthma, the prevalence of asthma were 5.5% (453), 5.9% (474), 5.1% (412), and there was significant difference between the two groups, the value of χ(2) were 130.522, 59.929 and 293.997, respectively (all P values < 0.05). Logistic regression analysis showed that passive smoking (OR = 1.7, 95%CI: 1.2 - 2.4), house decoration (OR = 1.5, 95%CI: 1.1 - 1.9) and pet (OR = 1.6, 95%CI: 1.1 - 2.3) were statistically significant to asthma in non-susceptible children. While passive smoking (OR = 1.3, 95%CI: 1.0 - 1.7) and house decoration (OR = 1.4, 95%CI: 1.1 - 1.7) were increased the risk of asthma.

CONCLUSIONIndoor air pollution is a risk factor of children' s asthma. Family history of asthma and physical susceptible children are high risk to asthma, and susceptible children are easily influenced by other risk factors.

Air Pollution, Indoor ; adverse effects ; analysis ; Asthma ; epidemiology ; etiology ; Child ; China ; epidemiology ; Environment ; Female ; Humans ; Male ; Risk Factors

BACKGROUND: As a special source of stem cells, dental pulp stem cells (DPSCs) make much progress in the development of tissue engineering field due to their high proliferation and self-renewal ability. In the certain conditions DPSCs can be induced to differentiate into a variety of specialized tissue cells, providing a new way for tissue engineering development. OBJECTIVE: To review the main progress in the DPSCs biological characteristics, original source, isolation method, and its related application in tissue engineering research. METHODS: "Dental pulp stem cell, differentiation, regenerative medicine, tissue engineering" in English and Chinese were termed as the keywords to search relevant articles about DPSCs and tissue engineering published from 2005 to 2017 in PubMed, Medline, WanFang, and CNKI databases. After removal of repetitive or irrelevant articles, 66 articles were finally reviewed. RESULTS AND CONCLUSION: With the development of tissue engineering and regenerative medicine, the effective combination of DPSCs and tissue engineering scaffolds have be further achieved. Recent studies on DPSCs focus on the properties of DPSCs differentiating into odontoblasts and osteocytes/osteoblasts and on the potential of nerve repair, vascular remodeling, corneal reconstruction and chondrogenic differentiation.

OBJECTIVETo evaluate the interaction effects of breastfeeding and passive smoking on asthma and asthma related symptoms among children.

METHODSUsing a cluster random sampling method, 2 elementary schools and 1 kindergarten were randomly selected from 7 cities of Liaoning province. The resulting 25 elementary schools and 50 kindergartens were included, and 31 049 children from the selected schools living up to 2 years were recruited in this survey. The information about the children's type of feeding up, living environment, passive smoking exposure, respiratory diseases and symptoms were collected. The interaction effects of breastfeeding and passive smoking on asthma and asthma related symptoms (persistent cough,persistent phlegm, current wheeze and allergic rhinitis) were evaluated with Glimmix procedure.

RESULTSThere were 31 049 children involved in this investigation. The age was (8.32 ± 2.75) years old. There were 23 987 (77.26%) children with breastfeeding and 11 820 (38.07%) children with passive smoking. The prevalence of asthma and allergic rhinitis were 6.22%(1491/23 987), 4.67%(1120/23 987) in children with breastfeeding, and were 7.70%(544/7062), 5.48%(387/7062) in children without breastfeeding,compared to the children without breastfeeding, the children with breastfeeding had lower risk of asthma(OR = 0.79, 95%CI:0.72-0.88) and allergic rhinitis(OR = 0.85, 95%CI:0.75-0.95); The prevalence of current wheeze was 7.89%(929/11 770) in children with father smoking, and was 5.37%(1036/19 279) in children without father smoking, compared to the children without father smoking, the children with father smoking increased the risk of current wheeze(OR = 1.51, 95%CI:1.38-1.65). The prevalence of persistent cough was 18.96%(51/269) in children with mother smoking, and was 9.51%(2926/30 780) in children without mother smoking,compared to the children without mother smoking, the children with mother smoking increased the risk of persistent cough(OR = 2.23, 95%CI:1.64-3.03). The prevalence of persistent phlegm was 5.69%(871/5316) in children with anyone smoking, and was 3.50%(550/15 733) in children without anyone smoking, compared to the children without anyone smoking, the children with anyone smoking increased the risk of persistent phlegm(OR = 1.67, 95%CI:1.49-1.86).Glimmix procedure analysis showed there was a significant interaction effects between breastfeeding and passive smoking. The estimated OR for father smoking among breastfeeding children were consistently lower than those among non-breastfeeding children for asthma. The estimated OR for mother smoking among breastfeeding children were consistently lower than those among non-breastfeeding children for allergic rhinitis. The estimated OR for anyone smoking among breastfeeding children were consistently lower than those among non-breastfeeding children for asthma and allergic rhinitis(all P values < 0.05).

CONCLUSIONBreastfeeding decreases the detrimental effects of passive smoking on asthma and asthma related symptoms in children.

Adolescent ; Asthma ; epidemiology ; Breast Feeding ; Child ; Child, Preschool ; Female ; Humans ; Male ; Risk Factors ; Smoking ; Surveys and Questionnaires ; Tobacco Smoke Pollution ; adverse effects

OBJECTIVETo investigate the association of single nucleotide polymorphisms (SNPs) in VEGF gene with the risk of endometriosis and adenomyosis.

METHODSGenotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 344 endometriosis patients, 174 adenomyosis patients, 360 frequency-matched control women of endometriosis and 199 frequency-matched control women of adenomyosis.

RESULTSNo significant difference was found in allele frequencies and genotype distributions of the -460C/T polymorphism between patients (endometriosis and adenomyosis) and control women (all P value > 0.05). However, there were significant differences in genotype and allele distributions of the VEGF -1154G/A polymorphism between patients (endometriosis and adenomyosis) and control women (all P value < 0.05). The genotype frequencies of the VEGF -1154 AA, GA, and GG in endometriosis patients and control women were 1.7%, 28.8%, 69.5% and 5.8%, 32.8%, 61.4%, respectively; and the A and G allele frequencies in the two groups were 16.1%, 83.9% and 22.2%, 77.8%, respectively. The genotype frequencies of the VEGF -1154 AA, GA, and GG in adenomyosis patients and control women were 2.9%, 23.6%, 73.6% and 7.0%, 34.2%, 58.8%, respectively; and the A and G allele frequencies in the two groups were 14.7%, 85.3% and 24.1%, 75.9% respectively. Compared with GA+ AA genotype, GG genotypes could significantly increase the risk of endometriosis (OR:1.43,95%CI:1.05-1.96) and adenomyosis (OR:1.95,95%CI:1.26-3.03).

CONCLUSIONThe VEGF -1154G/A polymorphism was associated with susceptibility to endometriosis and adenomyosis, and the GG genotype could significantly increase the risk of developing endometriosis and adenomyosis. However, the VEGF -460C/T polymorphism was not associated with susceptibility to endometriosis and adenomyosis in the population studied.

5' Untranslated Regions ; Adult ; Biophysical Phenomena ; Endometriosis ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo synthesize two antigens-Ag85b and HspX of Mycobacterium tuberculosis H37Rv with molecular biological methods and to observe their biologic activity after co-administration of adjuvants (aluminum and/or CpG) in mice.

METHODSRecombinant expression plasmids pET30a-Ag85b and pET30a-HspX were constructed. The objective DNA fragments was characterized with restriction enzyme. Then the recombinant plasmids were transformed into E. coli BL-21, and two proteins were expressed by induction of isopropyl beta-D-1-thiogalactopyranoside. After purification with anion exchange column Source30, QHP, and hydrophobic chromatography column, two proteins were identified by amino acid sequencing. After the successful preparation of these two antigens, they were co-administered in mice with adjuvants of aluminum and/or CpG (Ag85b, Ag85b + Al, Ag85b + CpG, Ag85b + Al + CpG; HspX, HspX + Al, HspX + CpG, HspX + Al + CpG); one group received normal saline and served as the control. Splenic lymphocytes were isolated for enzyme-linked immunosorbent spot assay to detect the secreted specific interferon-gamma (IFN-gamma); in addition, lymphocytes proliferation test was performed to observe lymphocytes proliferation after in vitro stimulated with two antigens.

RESULTSThe purity of two proteins reached 95% after purification. The N-terminal amino acid sequence (15 aa) of the purified proteins was same as the target sequence. For Ag85b, the secreted specific IFN-gamma from isolated splenic lymphocytes after having been stimulated in vitro with Ag85b (80 microg/ml) remarkably increased in Ag85b + CpG group, Ag85b + Al group, and Ag85b + CpG + Al group; the changes were significantly different between these three groups and control group (P < 0.05). For HspX, the changes were significantly different between HspX + Al + CpG group and normal sodium group, although remarked increase of IFN-gamma was also observed in HspX group, HspX + Al group, and HspX + CpG group.

CONCLUSIONSAg85b and HspX were successfully expressed and purified. A cell-mediated immunity may be induced when the antigens are co-administered with adjuvants of aluminum and/or CpG in mice, indicating that the recombinant proteins are bioactive.

Acyltransferases ; administration & dosage ; isolation & purification ; therapeutic use ; Adjuvants, Immunologic ; therapeutic use ; Animals ; Antigens, Bacterial ; administration & dosage ; isolation & purification ; therapeutic use ; Bacterial Proteins ; administration & dosage ; isolation & purification ; therapeutic use ; Escherichia coli ; Immunity, Cellular ; Interferon-gamma ; Mice ; Mycobacterium tuberculosis ; immunology ; metabolism ; Recombinant Proteins ; administration & dosage ; isolation & purification ; therapeutic use

To determine heat-shock protein (Hsp)90 expression is connected with cellular apoptotic response to heat stress and its mechanism, chicken (Gallus gallus) primary myocardial cells were treated with the Hsp90 promoter, aspirin, and its inhibitor, geldanamycin (GA), before heat stress. Cellular viability, heat-stressed apoptosis and reactive oxygen species level under different treatments were measured, and the expression of key proteins of the signaling pathway related to Hsp90 and their colocalization with Hsp90 were detected. The results showed that aspirin treatment increased the expression of protein kinase B (Akt), the signal transducer and activator of transcription (STAT)-3 and p-IKKα/β and the colocalization of Akt and STAT-3 with Hsp90 during heat stress, which was accompanied by improved viability and low apoptosis. GA significantly inhibited Akt expression and p-IKKα/β level, but not STAT-3 quantity, while the colocalization of Akt and STAT-3 with Hsp90 was weakened, followed by lower cell viability and higher apoptosis. Aspirin after GA treatment partially improved the stress response and apoptosis rate of tested cells caused by the recovery of Akt expression and colocalization, rather than the level of STAT-3 (including its co-localization with Hsp90) and p-IKKα/β. Therefore, Hsp90 expression has a positive effect on cellular capacity to resist heat-stressed injury and apoptosis. Moreover, inhibition of Hsp90 before stress partially attenuated its positive effects.
Apoptosis* ; Aspirin ; Cell Survival ; Chickens* ; Heat Stress Disorders ; Heat-Shock Proteins* ; Hot Temperature* ; HSP90 Heat-Shock Proteins ; In Vitro Techniques* ; Proto-Oncogene Proteins c-akt ; Reactive Oxygen Species ; Transducers

BACKGROUND: Morinda officinalis oligosaccharide is the main active ingredient of morinda officinalis extract,which can promote the angiogenesis of ischemic tissue, but the mechanism is unknown. At present, there are two ways for endothelial repair:vascular endothelial cell division or differentiation from vascular endothelial progenitor cells in the peripheral blood. Here, we attempted to explain the pro-angiogenesis mechanism of morinda officinalis oligosaccharide by exploring whether there is a correlation between morinda officinalis oligosaccharide and the biological function of vascular endothelial progenitor cells, thereby providing experimental reference for new drug development. OBJECTIVE: To observe the effect of morindae officinalis oligosaccharide on the proliferation, differentiation and paracrine of vascular endothelial progenitor cells. METHODS: Vascular endothelial progenitor cells were isolated from healthy human peripheral blood, and divided into two groups: control group (without morindae officinalis oligosaccharide) and experimental group (with 0.15 g/L morindae officinalis oligosaccharide), followed by 48 hours of in vitro culture.The proliferation of vascular endothelial progenitor cells was tested by fluorescent staining;the ratio of vascular endothelial progenitor cells expressing CD31 was detected by flow cytometry; and the levels of vascular endothelial growth factor, stromal cell-derived factor 1 and interleukin 8 were analyzed by enzyme-linked immunosorbent method. RESULTS AND CONCLUSION: The percentage of vascular endothelial cells expressing CD34, CD133 or VEGFR- 2 was (84.72±4.34)%. After 48 hours of culture by 0.15 g/L morindae officinalis oligosaccharide, the proliferation rate and the positive expression of CD31 in the experimental group were significantly higher than those in the control group (P < 0.05). The levels of vascular endothelial growth factor, stromal cell-derived factor 1 and interleukin 8 in the experimental group were also higher than those in the control group (P < 0.05). To conclude, morindae officinalis oligosaccharide can promote the proliferation and differentiation of vascular endothelial progenitor cells, and meanwhile, it can stimulate the release of vascular endothelial growth factor, stromal cell-derived factor 1 and interleukin 8 from vascular endothelial progenitor cells through the paracrine pathway. Consequently, it is a potential drug for myocardial ischemic diseases.