In this study, we investigated the effect of Cigu Xiaozhi formula on HSC-T6 activity in hypoxic microenvironment based on network pharmacology and computer-aided drug design, and predicted and verified its possible targets and related signaling pathways. The potential active components and targets of Cigu Xiaozhi formula were screened by searching Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Encyclopaedia of Traditional Chinese Medicine (ETCM) and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) databases, and the liver fibrosis related targets retrieved from Gene Cards and Pharm GK database were integrated to obtain the potential targets of Cigu Xiaozhi formula in the treatment of liver fibrosis. GO enrichment analysis and KEGG signaling pathway enrichment analysis were performed on Omic Share platform, and Cytoscape software was used to construct the "potential active ingredient-key target-pathway" network. The active components and target proteins were subjected to molecular docking analysis by Auto Dock software. According to the results of molecular dynamics simulation and binding free energy calculation, the top 5 active components with degree were scored. The active components stigmasterol and β-sitosterol were subjected to molecular docking. CoCl₂ was used to induce HSC-T6 cells to construct hypoxia model in vitro. The cell viability was detected by CCK-8 assay, and the optimal time and concentration of hypoxia model of HSC-T6 cells was determined to be 100 µmol·L^-1 CoCl₂ for 24 h. Under hypoxia condition, HSC-T6 cells were activated, the wound healing rate was significantly increased, and the fluorescence signal of activation marker protein α-smooth muscle actin (α-SMA) was significantly enhanced. However, 6% drug-containing serum could inhibit the activation of HSC-T6 cells, and the wound healing rate was significantly decreased, and the fluorescence signal of α-SMA was significantly weakened. Further studies showed that the expressions of hypoxia-inducible factor-1α (HIF-1α), α-SMA and key proteins of Hedgehog (Hh) signaling pathway in HSC-T6 cells were up-regulated under hypoxia, while the expressions of HIF-1α, α-SMA, Patched-1 (Ptch-1) and glioma related oncogene homology-1 (Gli-1) were down-regulated in 6% drug-containing serum group, the YC-1 group and the cyclopamine group. These results indicated that HIF-1α and Hh signaling pathways were involved in the activation of HSC-T6 cells, and the traditional Chinese medicine Cigu Xiaozhi formula could inhibit the activation of HSC-T6 cells, and the mechanism may be related to the inhibition of HIF-1α expression and the blocking of Hh signaling pathway. In conclusion, Cigu Xiaozhi formula can inhibit the activation of HSC-T6 cells by directly acting on HIF-1α and Hh signaling pathway, and exert an anti-hepatic fibrosis effect. The animal experimental protocol has been reviewed and approved by Laboratory Animal Ethics Committee of Gansu University of Chinese Medicine, in compliance with the Institutional Animal Care Guidelines.

OBJECTIVETo study the distribution and drug resistance of the isolated bacteria from children with acute respiratory infection (ARI) in Dalian.

METHODSBetween January 2006 and February 2007, 930 children with ARI were enrolled, including 364 with acute upper respiratory infection (AURI), and 566 with acute lower respiratory infection (ALRI). The AURI children, who did not receive antimicrobial agent treatment or received oral antimicrobial agents 1-2 times, had bacterial cultures of pharyngeal swab. The ALRI children, who received intravenous antibacterial agents more than 3 days, had bacterial cultures of sputum and bronchoalveolar lavage fluid (BALF). Isolated bacteria were identified by the ATB system (Bio-Merieux, France). Antimicrobial susceptibility testing was carried out by means of Kirby-bauer.

RESULTSA total of 404 isolates (43.4%) were identified. Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 22.5%, 12.1% and 7.4% respectively. In the isolates from AURI, Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 43.9%, 22.0% and 9.1% respectively; Escherichia coli, Klebsiella pneumonia and Nonfermenters accounted for 4.5%, 8.3% and 3.0% respectively. In the isolates from ALRI, Haemophilus influenzae, Haemophilus parainfluenzae and Streptococcus pneumoniae accounted for 12.1%, 7.4% and 6.6% respectively; Escherichia coli, Klebsiella pneumoniae and Nonfermenters accounted for 16.9%, 13.2% and 21.8% respectively. The resistant rates of Haemophilus to ampicillin and TMP-SMZ were 29.3% and 32.9% respectively, and to amoxicillin-clavulanic acid, cefalotin, cefaclor, cefuroxime and cefotaxime were 12.1%, 10.0%, 10.0%, 11.4% and 5.7%, respectively. The resistant rate of Haemophilus to ampicillin, amoxicillin-clavulanic acid, cefaclor, tetracycine and TMP-SMZ in the ALRI group were significantly higher than that in the AURI group (P<0.05 or 0.01).

CONCLUSIONSIn Dalian, Haemophilus was the main isolate of children with ARI. The distribution of bacteria was different between ALRI and AURI. In ALRI, Gram-negative bacilli were in a higher proportion, and the resistant rates of Haemophilus influenzae and Haemophilus parainfluenzae to ampicillin, amoxicillin-clavulanic acid and cefaclor were higher.

Acute Disease ; Adolescent ; Bacteria ; drug effects ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Male ; Respiratory Tract Infections ; drug therapy ; microbiology

Objective To investigate the influence of high intensity interval exercise (HIIT) on peroxidation and vascular endothelial function for experimental hyperhomocysteinemia (HHcy) rats. Methods Thirty five male rats were randomly divided into 4 groups. Control group (n=8) was given ordinary feed. High methionine group (n=27) was given 3% methionine on this basis, and divided into model group, folic acid group and HIIT+ folic acid group, with 9 rats per group for 16 weeks. Serum homocysteine (Hcy) , content of plasma malondialdehyde (MDA) , hydroxyl radical (OH-), total antioxidant capacity (T-AOC), activity of glutathione peroxidase (GSH-PX) and superoxide dismutase (SOD) were measured, as well as the level of Nitric Oxide (NO), Nitric Oxide Synthase (NOS) and Endothelin 1 (ET-1) . The pathology of abdominal aortas was analyzed.Results Sixteen weeks after intervention, there was no significant difference between HIIT + folic acid group and the control group (P>0.05) . The levels of serum Hcy in the model group, folic acid group and the HIIT+folic acid group were (23.95±3.35) μmol/L,(8.73±0.60) μmol/L, and (6.19±0.34) μmol/L respectively (P<0.05) . Sixteen weeks after intervention, the content of MDA in HIIT+ folic acid group reduced, and there was no significant difference compared with the control group (P>0.05). The level of SOD and GSH-PX increased in HIIT+ folic acid group and folic acid group, and there was a significant difference compared with the model group. There were significant differences in activities of SOD and GSH-PX in HIIT+ folic acid group when compared with folic acid group (P<0.05). Compared with the control group, there were significant differences in levels of ET-1, NOS and NO in folic acid group (P<0.05), while there was no significant difference in the level of ET-1 and NOS between HIIT+folic acid group and control group (P>0.05) . Mild atherosclerotic lesions were observed in the HIIT+folic group. Conclusion High methionine diet can reduce the level of serum Hcy in HHcy rats, and high intensity interval exercise combined with folic acid intervention could reduce the level of serum Hcy, improve oxidative stress state, reduce the injury of endothelial function, and thus to alleviate atherosclerotic lesion.

Objective: To observe the effects of electroacupuncture (EA) on the protein and gene expressions of Bax, Caspase-3 and Bcl-2 in cerebral cortex of type 2 diabetic rats with cognitive impairment (CI), and to explore the mechanism of EA in improving the learning and memory abilities. Methods: A total of 100 Sprague-Dawley (SD) rats were divided into a normal group (n=10) and a model group (n=90) by random number table method. Rats in the model group were intraperitoneally injected with a small dose of streptozotocin (STZ) to establish the type 2 diabetic models, after being fed with high-fat and high-sugar diet for 1 month. Twenty CI rats were selected from the 50 successful model rats by the Morris water maze (MWM) test and randomly divided into a model group and an EA group according to the blood glucose level and MWM data (n=10). Rats in the EA group received acupuncture at Zusanli (ST 36), Neiting (ST 44) and Yishu (Extra), of which Zusanli (ST 36) and Neiting (ST 44) were stimulated by EA apparatus, 20 min/time, once a day for 6 d a week and 4 consecutive weeks. The rats in the model and the normal groups were fixed without treatment. After 4-week treatment, the random blood glucose level of the rats was measured; the learning and memory abilities of rats were measured by MWM; terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect apoptotic cells; Western blot (WB) and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the protein and gene expressions of Bax, Caspase-3 and Bcl-2 in cerebral cortex. Results: After modeling, the random blood glucose level and the escape latency tested by MWM were significantly increased, and the number of crossing the platform tested by the MWM was decreased in the EA and model groups, and were significantly different from those in the normal group (P<0.05 or P<0.01), while the differences between the model group and the EA group were not statistically significant (all P>0.05). After 4-week treatment, the random glucose level and the escape latency tested by MWM were significantly increased (both P<0.05), and the number of crossing the original platform tested by the MWM was significantly reduced (P<0.01), the protein and gene expressions of Bax and Caspase-3 were significantly increased (all P<0.001), the protein and gene expressions of Bcl-2 were significantly reduced (both P<0.001), and the number of neuron apoptosis was significantly increased (P<0.001) in the model group than in the normal group; the random blood glucose level was significantly reduced (P<0.05), the escape latency tested by MWM was significantly shortened (P<0.05), and the number of crossing the original platform tested by MWM was significantly increased (P<0.05), the protein and gene expressions of Bax and Caspase-3 were significantly reduced (all P<0.001), the protein and gene expressions of Bcl-2 were significantly increased (both P<0.001), and the number of neuron apoptosis was significantly reduced (P<0.001) in the EA group than in the model group. Conclusion: EA can improve the learning and memory damages induced by type 2 diabetic model rats with CI; the action mechanism may be achieved via anti-apoptosis.

OBJECTIVETo study the variation and evolution rules and analyze the genetic characterization of echovirus 30 during an outbreak of aseptic meningitis.

METHODSFor all the echovirus 30 isolates during the outbreaks in Zhangqiu city in 2003, complete VP1 genes were determined. Phylogenetic tree reconstruction and pairwise sequence were determined to compare the homological relationship and evolution rules between Zhangqiu strains and the other reported echovirus 30 in GenBank.

RESULTSPairwise sequence comparisoms in VP1 region indicated that there were 98.9%-99.5% identity of nucleotide acid among Zhangqiu strains, and having the highest homology with Taian strains (98.5%-99.0%) isolated in Shandong province. At the same time, when comparing with Zhejiang and Jiangsu strains, the nucleotide homology showed 98.2%-98.9%, relatively low identity with Taiwan strains (87.4%-87.6%) and French strains (89.1%-89.7%) but only 82.4%-82.8% identity when comparing with echovirus 30 prototype strain Bastianni. Data from phylogenetic tree analyses indicated that all the echovirus 30 correlated with aseptic meningitis in China recently came from the same evolution linkage and formed a monophyletic cluster.

CONCLUSIONIn 2003, there was an circulation of echovirus 30 in Shandong province which causing an endemic of aseptic meningitis in local areas. Data from phylogenetic tree analyses indicated that the antigenic characteristics of echovirus 30 in mainland China might be different from strains early isolated in other continents and formed a new genotype.

China ; epidemiology ; DNA, Viral ; analysis ; Disease Outbreaks ; Enterovirus B, Human ; classification ; genetics ; Enterovirus Infections ; epidemiology ; Genotype ; Humans ; Meningitis, Aseptic ; Phylogeny ; Sequence Analysis, DNA

Adult ; Electrocardiography ; Humans ; Male ; Myotonic Dystrophy ; complications ; pathology ; physiopathology ; Syringomyelia ; complications ; pathology ; physiopathology

OBJECTIVETo study the effects of prolonged 75% oxygen exposure on the expression of vascular endothelial growth factor (VEGF) and its receptors (VEGFR1 and VEGFR2) in the neonatal rat lungs and to elucidate the effects of prolonged exposure of high concentration of oxygen on lung vascular development and its relationship with bronchopulmonary dysplasia (BPD).

METHODSForty eight Sprague-Dawley rat pups were randomly exposed to air (control group) and 75% oxygen (experimental group) 12 hrs after birth. The rats were sacrificed 7, 14 and 21 days after exposure and their lungs were sampled. The lung sections were stained with hematoxylin and eosin for histological evaluation. Expression of VEGF, VEGFR1 and VEGFR2 protein and mRNA was detected by immunohistochemistry and RT-PCR.

RESULTSAfter being exposed to 75% oxygen for 21 days, lung tissues had pathological changes as 'new' BPD. Expressions of VEGF protein (10.9 + or - 2.7 vs 30.8 + or - 6.4), VEGFR1 protein (5.4 + or - 1.4 vs 15.6 + or - 3.4) and VEGFR2 protein (11.3 + or - 2.6 vs 21.7 + or - 4.5) on day 21 in the experimental group decreased significantly as compared with the control group (p<0.05). The expression of VEGF mRNA (1.6 vs 3.3), VEGFR1 mRNA (0.4 vs 6.6) and VEGFR2 mRNA (0.5 vs 4.9) on day 21 in the experimental group also decreased significantly as compared with the control group (p<0.05).

CONCLUSIONSProlonged exposure of high concentration of oxygen may cause BPD possibly by inhibiting lung vascular development in neonatal rats.

Animals ; Animals, Newborn ; Bronchopulmonary Dysplasia ; etiology ; Female ; Humans ; Infant, Newborn ; Lung ; blood supply ; Male ; Oxygen ; toxicity ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; analysis ; genetics ; Vascular Endothelial Growth Factor Receptor-1 ; analysis ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; analysis ; genetics

The distribution information of Cistanche deserticola was collected by interview investigation and field survey, and 55 related environmental factors were collected, the habitat suitability study was conducted based on geographic information system (GIS) and Maximum entropy model. The AUCs of ROC curve were both above 0.9, indicating that the predictive results with the maxent model were highly precise. The results showed that 14 major environmental factors have obvious influence on ecology suitability distributions of C. deserticola, including vegetation type et al, the suitable distribution areas are mainly concentrated in the central of Alxa Youqi, the north of Alxa Zouqi and the south-east of Ejin Banner, including Tamusu towns, Alateng towns et al, The zoning results basically coincide with the genuine producing areas, and further afford new suitable distribution areas, which can provide reference for the siting of introduction and cultivation of C. deserticola.
China ; Cistanche ; growth & development ; Ecosystem ; Environment ; Geographic Information Systems ; Rain ; Soil ; chemistry ; Temperature

OBJECTIVETo study the therapeutical effects of Shuangshen Ningxin capsule on miniature swine after myocardial ischemia by intervention.

METHODMyocardial ischemic model miniature swine induced by self-thrombus via cardiac catheter in left anteriar descending coronary artery (LAD), were administrated Shuangshen Ningxin capsule for 6 days. The changes of coronary arteriography, hemodynamics, biochemistry and pathohistology were observed.

RESULT6 days after modeling, LAD in myocardial ischemic miniature swine was basically embolized, cardiac output (CO), cardiac index (CI), left cardiac work (LCW) and left cardiac work index (LCWI) obviously lowed, and pathohistological analysis revealed myocardial degeneration, necrosis, fibrosis and inflammatory cell infiltration. After being administered with shuangshen Ningxin capsule 6 days, the degree of self-thrombus blocked LAD reduced, hemodynamic indexes of CO, CI, LCW, LCWI and blood plasm superoxide dismutase (SOD) activity increased, and systemic vascular resistance (SVR), systemic vascular resistance index (SVRI) and malondialdehyde (MDA) content were lowed. on the same time, pathohistological degeneration and necrosis reduced.

CONCLUSIONShuangshen Ningxin capsule has anti-myocardial ischemia effect by improving cardiac muscle systolic function, increasing left cardiac work, inhibiting cardiac muscle cellular membrane lipid peroxidation.

Animals ; Capsules ; Cardiac Output ; drug effects ; Cardiotonic Agents ; administration & dosage ; pharmacology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Hemodynamics ; drug effects ; Male ; Malondialdehyde ; blood ; Myocardial Contraction ; drug effects ; Myocardial Ischemia ; blood ; physiopathology ; prevention & control ; Myocardium ; pathology ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Salvia miltiorrhiza ; chemistry ; Superoxide Dismutase ; blood ; Swine ; Swine, Miniature ; Vascular Resistance ; drug effects

OBJECTIVETo investigate the influence of partial deletions in the AZFc region of the Y chromosome on spermatogenesis.

METHODSWe selected 9 sequence tagged sites (sY1258, sY1291, sY254, sY255, sY1201, sY1206, sY1161, sY1197 and sY1191) in the AZFc region of the Y chromosome, with ZFX/ZFY and SRY (sY14) as the interior control. We amplified by multiplex PCR the DNA of 160 patients with azoospermia or severe oligozoospermia that showed no microdeletion of the Y chromosome (the case group) and another 76 males with normal fertility (the control group). For the individuals suspected of DAZ gene deletion, we detected the single nucleotide polymorphism sites (SNPs) in the four copies of the DAZ gene by single nucleotide variation (SNV) analysis to determine the types of DAZ copy deletion.

RESULTSIn the case group, there were 10 cases of gr/gr (sY1291) deletion (6.3%), 14 b2/b3 (sY1191) deletion (8.8%), 1 sY1291,sY1197 deletion (0.6%), 1 b1/b2 deletion (0.6%) and 1 b1/b3 deletion (0.6%), while in the control group, there were 4 cases of gr/gr deletion (5.3%) and 4 b2/b3 deletion (5.3%). SNV analysis showed DAZ1/DAZ2 deletion in all those with gr/gr and those with b1/b3 deletion, DAZ3/DAZ4 deletion in those with b2/b3 deletion, and DAZ-SNV sY587 deletion in 1 case of sY1291, sY1197 deletion, but no DAZ deletion was found in 1 case of b1/b2 deletion.

CONCLUSIONB2/b3 (sY1191) and gr/gr (sY1291) deletions are genomic polymorphisms and quite common in the normal Chinese population; while b1/b2, b1/b3, and sY1291, sY1197 deletions may be high risk factors of dyszoospermia.

Case-Control Studies ; Chromosomes, Human, Y ; Humans ; Male ; Oligospermia ; genetics ; Sequence Deletion ; Spermatogenesis