OBJECTIVE: To investigate the effects of different intensity intermittent exercise on the body function of obese rats, and to provide basis for the prevention and treatment of obesity. METHODS: Eighty SD rats were randomly divided into normal diet group (n=20) and high-fat diet group (n=60). After adaptive feeding for 8 weeks, 8 normal diet rats and 32 high-fat obese rats were selected for follow-up experiments. The experimental rats were randomly divided into 5 groups (n=8): control diet-sedentary (CS),with ordinary feed and without any exercise; high diet-sedentary (HS), with high-fat feed feeding and without any exercise; high diet-continual exercise(HC), 60 min/day,5 days/week with 6 weeks; high diet-long time-low frequency interval exercise(HLL), 30 min/time,twice/day (intermittent 6 h), 5 days/week with 6 weeks; high diet-short time-high frequency interval exercise(HSH), 20 min/ time, 3 times/d (intermittent 3 h), 5 days/week with 6 weeks. The training intensity of rats in each exercise group was 25 m/min. After 6 weeks, rats in each groups were weighed, and resting metabolic rate(RMR), fasting blood glucose(FBG), triglyceride(TG) and other biochemical indexes were detected, and fat and muscle weight were measured. RESULTS: Before experiment, there were no significant differences in RMR, FBG and TG in each groups(P＞0.05).The body weight of HSH, HLL, HC and HS groups was higher than that of CS group (P＜0.05). After the experiment, RMR of the HSH,HLL and HC groups was significantly higher than that of HS and CS groups (P＜0.05), but without significant difference among the HSH,HLL and HC groups (P＞0.05).The body weight of HSH, HLL and HC groups was significantly lower than that of HS group (P＜0.05), but the three groups was not significant (P＞0.05); perirenal fat(PF), idymis fat(EF), perirenal fat/weight(PF/W) and epididymis fat/weight(EF/W) of HSH, HLL, HC group were significantly lower than those of HS group (P＜0.01), while there was no statistical difference among the three groups (P＞0.05). There was no significant difference in gastrocnemius(GM) and quadricep(QF) of each group (P＞0.05), gastrocnemius/weight(GM/W) and quadriceps/weight (QF/W) in HSH,HLL and HC groups were higher than those of HS group (P＜0.05),while there was no significant difference among HSH,HLL and HC groups (P＞0.05);FEB,TG of HSH,HLL,HC group were lower than those of CS and HS group (P＜0.05),but the difference with HS group was more significant (P＜0.01),there was no significant difference among training groups(P＞0.05). CONCLUSION 6 weeks of intermittent exercise of different intensity had a good intervention effect on the body composition of obese rats, and high diet-short time-high frequency interval exercise (HSH) may be more effective.
Animals ; Body Composition ; Body Weight ; Diet, High-Fat ; Male ; Obesity ; therapy ; Physical Conditioning, Animal ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Human beta defensin 2 (HBD-2) may play an important role in human defense against infection. Its antimicrobial capacity has been fully documented in in vitro study. In order to evaluae its in vivo effects, we developed an HBD-2 transgenic mouse model. The HBD-2 minigene containing CMV promoter, full length of HBD-2 cDNA and BGH polyA tail was generated by PCR amplification and introduced into the fertilized oocytes of C57 X ICR hybridized mouse by microinjection, and offspring were produced. DNA was isolated from the tails of the mouse pups, and the HBD-2 minigene incorporation was analyzed by PCR using HBD-2 specific primers. The HBD-2 gene expression in the multi-tissues of transgenic mice was determined at mRNA level by RT-PCR and at peptide level by immunohistological staining with the use of HBD-2 monoclonal antibody. The results showed that among 17 F0 transgenic mice, HBD-2 positive signal was determined by PCR in 4 mice, suggesting that HBD-2 minigene has been incorporated into the offspring mice. Meanwhile, a widespread expression of HBD-2 mRNA and peptide was detected in the F1 transgenic mice's multi-tissues such as trachea, lung, intestine, esophagus, testis, spleen, skin, endothelium and brain.
Animals ; Anti-Infective Agents ; Humans ; Mice ; Mice, Inbred C57BL ; Mice, Inbred ICR ; Mice, Transgenic ; Models, Animal ; Polymerase Chain Reaction ; RNA, Messenger ; analysis ; biosynthesis ; genetics ; beta-Defensins ; biosynthesis ; genetics

OBJECTIVETo study the effects of benzo(a)pyrene (BaP) on the cell cycle distribution and activities of mitogen-activated protein kinase (MAPK) signal molecules (ERK1/2, JNK1/2 and p38) in human embryo lung cells (HELF), and to investigate the relationship between alterations of MAPK protein phosphorylation and the cell cycle distributions.

METHODSThe phosphorylation of MAPK were induced by exposing HELF cells to BaP at 0.1, 0.5, 2.5 and 12.5 micromol/L. The phosphorylation and protein expression levels of ERK1/2, JNK1/2 and p38 were determined through western-blotting assay. And the flow cytometry assay was used to measure the cell cycle effects in HELF cells after treatment with 2.5 micromol/L BaP for 24 h.

RESULTSThe phosphorylation levels of ERK1/2, JNK1/2 and p38 were significantly increased through BaP exposure. In addition, the phosphorylation of these three MAPKs has similar alteration pattern. We found that exposure of cells to 2.5 microM of BaP for 24 h resulted in a decrease of G(0) and G(1) population by 11.9% (F = 41.38, P < 0.01) and an increase of S population by 17.2% (F = 68.13, P < 0.01). Three chemical inhibitors of MAPK (AG126, SP600125 and SB203580) could significantly inhibit the cell cycle alteration because of BaP treatment.

CONCLUSIONERK1/2, JNK1/2 and p38 could positively regulate the BaP independently induced cell cycle alterations.

Benzo(a)pyrene ; toxicity ; Cell Cycle ; drug effects ; Cells, Cultured ; Fibroblasts ; drug effects ; metabolism ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Lung ; cytology ; embryology ; MAP Kinase Kinase 4 ; metabolism ; MAP Kinase Signaling System ; drug effects ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Mitogen-Activated Protein Kinase 8 ; metabolism ; Mitogen-Activated Protein Kinase 9 ; metabolism ; Signal Transduction ; drug effects ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo investigate the effects of citrus reticulata blanco extract on the proliferation and collagen metabolism of fibroblasts from human hypertrophic scar.

METHODSHuman hypertrophic scar fibroblasts from two burn patients obtained from plastic surgery were cultured in vitro and divided into experimental group (n = 12, with basic culture medium and 2.5, 5.0, 10.0,25.0 mg/L citrus reticulata blanco extract, respectively, 3 bottles for each concentration of citrus reticulata blanco extract ), control group 1 (n = 3, with basic culture medium) , and control group 2 ( n = 3, with basic culture medium and 5% ethyl alcohol). The cell proliferation in each group was observed with MTT method, then the inhibition rate was calculated. Apoptosis and its index ( AI) in each group were determined after TUNEL staining . The changes in the content of ICTP and PINP in each group were observed by radioimmunity.

RESULTSThe inhibition rate in the experimental group with the citrus reticulata blanco extract in concentration of 2. 5, 5.0, 10.0, 25. 0 microg/ ml were (7. 100+/-0.038)% , (8. 100+/- 0. 048)% , (10. 900+/-0. 055)%, (15.900+/-0. 097) %, respectively, which were significantly higher than those in other two groups ( P <0.05 ). The Al (69. 7% , 71.7%, 86.4% , 95.2% ), ICTP [(17.2+/-0.6), (18.3+/-0.6), (19.8+/-0.5), (23.2+/-0.6) microg/L] and PINP [ (101.7+/-1.4) , (107. 8+/-1. 1) , (111.6+/-1.2) , (124. 6+/-1.3) microg/L] in experimental group with the citrus reticulata blanco extract in concentration of 2.5, 5.0, 10.0 , 25.0 mg/L were also obviously higher than other two control groups( P <0.05) ,but these indices in control 1 group were similar to those in control 2 group( P >0. 05).

CONCLUSIONThe citrus reticulata blanco extract might be beneficial for the management of hypertrophic scar through inhibition of the proliferation of fibroblasts in hypertrophic scar, by promoting apoptosis and collagen degradation.

Apoptosis ; drug effects ; Cell Division ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Citrus ; chemistry ; Collagen Type I ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; metabolism ; Humans

BACKGROUNDBoth uniportal and triportal thoracoscopic lobectomy and sublobectomy are feasible for early-stage non-small cell lung cancer (NSCLC). The aim of this study was to compare the perioperative outcomes of uniportal and triportal thoracoscopic lobectomy and sublobectomy for early-stage NSCLC.

METHODSA total of 405 patients with lung lesions underwent thoracoscopic lobectomy or sublobectomy through a uniportal or triportal procedure in approximately 7-month period (From November 2014 to May 2015). A propensity-matched analysis, incorporating preoperative variables, was used to compare the short-term outcomes of patients who received uniportal or triportal thoracoscopic lobectomy and sublobectomy.

RESULTSFifty-eight patients underwent uniportal and 347 patients underwent triportal pulmonary resection. The conversion rate for uniportal and triportal procedure was 3.4% (2/58) and 2.3% (8/347), respectively. The complication rate for uniportal and triportal procedure was 10.3% and 9.5%, respectively. There was no perioperative death in either group. Most patients had early-stage NSCLC in both groups (uniportal: 45/47, 96%; triportal: 313/343, 91%). Propensity score-matching analysis demonstrated no significant differences in operation time, intraoperative blood loss, numbers of dissected lymph nodes, number of stations of lymph node dissected, duration of chest tube, and complication rate between uniportal and triportal group for early-stage NSCLC. However, the duration of postoperative hospitalization was longer in the uniportal group (6.83 ± 4.17 vs. 5.42 ± 1.86 d, P = 0.036) compared with the triportal group.

CONCLUSIONSUniportal thoracoscopic lobectomy and sublobectomy is safe and feasible, with comparable short-term outcomes with triportal thoracoscopic pulmonary resection. Uniportal lobectomy and sublobectomy lead to similar cure rate as triportal lobectomy and sublobectomy for early NSCLC.

Adult ; Aged ; Blood Loss, Surgical ; Carcinoma, Non-Small-Cell Lung ; pathology ; surgery ; Female ; Humans ; Length of Stay ; Lung ; pathology ; surgery ; Lung Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Operative Time ; Pneumonectomy ; adverse effects ; methods ; Prospective Studies ; Thoracic Surgery, Video-Assisted ; adverse effects ; methods ; Treatment Outcome

Objective:To observe the effect of atorvastatin on the expression of soluble fractalkine in the circulation of patients with acute coronary syndrome .Methods:40 patients with acute coronary syndrome (ACS ) confirmed by coronary angiography were selected ,including 20 patients with acute myocardial infarction (AMI group) and20 patients with unstable angina pectoris (UAP group) .Another 20 hyperlipidemia patients with negative coronary angiography were selected as the control group .All three groups were given atorvastatin 20 mg per time ,one time per day for 3 months .Fractalkine levels in elbow vein blood were detected before and after treatment by ELISA method . CX3CR1 expression in peripheral blood mononuclear cells (PBMC) was examined by flow cytometry .Results:On admission ,fractalkine levels in the circulation of AMI group ([1047 ± 348 .9] pg/mL) and UAP Group ([777 .2 ± 324 .4] pg/mL) were significantly higher than that of the control group ([485 .1 ± 26 .09] pg /mL , P< 0 .001 , P= 0 .004);CX3CR1 expression rates of CD14+ monocytes in the circulation of AMI group ([38 .8 ± 15 .4]% ) and UAP group ([29 .2 ± 11 .9]% ) were significantly higher than that of the control group ([15 .2 ± 7 .9]% , P= 0 .009 , P= 0 .03) .After 3 months of atorvastatin treatment ,AMI ([740 .1 ± 269 .9] pg/mL) and UAP ([523 .8 ± 188 .5]pg/mL) fractalkine levels were significantly lower than before treatment (P= 0 .005 , P=0 .01 ) .Conclusions: Atorvastatin can significantly reduce fractalkine expression in the circulation of acute coronary syndrome patients ,and then achievethe immune effect of anti-atherosclerosis inflammation .

OBJECTIVETo develop a rapid duplex Real-time reverse transcription PCR (rRT-PCR) method to detect E119V mutation on neuraminidase (NA) of influenza A(H3N2) subtype with drug resistance to oseltamivir.

METHODSTwenty-six NA genes of influenza A(H3N2) virus between 2000 and 2012 in GenBank database were selected as the target genes, and specific TaqMan-MGB probe was designed to target the E119V amino acid change in neuraminidase protein. rRT-PCR was then performed and evaluated for the sensitivity, specificity and reproducibility using virus with E119V mutation and clinical samples.

RESULTSThis study described the validation of a highly sensitive and specific duplex rRT-PCR for detection of substitutions leading to the E119V amino acid change in NA protein of influenza A(H3N2). Fluorescence signals could be detected even when diluted a A (H3N2) virus (HA = 8) into 10(-5) and linear correlation between the logarithm of the viral titer with the Ct values was observed. In addition, the assay was highly specific in that there was no cross-react with other respiratory viruses, nor did two TaqMan-MGB probes. E119V substitution in quasispecies with both sensitive and resistant viruses could be detected as well. The limit of detection was 5% for quasispecies with high concentrations and 50% for quasispecies with low concentrations. The average coefficient of variation (CV) for within-run assays was 2.32% and 0.57% for H3N2-119E and H3N2-119V primer/probe sets separately, 1.77% and 0.97% for average CV of between-run assays, which exhibited good repeatability. Sequence analysis of twenty NA genes verified glutamic acid (E) at amino acid site 119, which was in consistent with the results from our rRT-PCR method.

CONCLUSIONThe assay developed in this study is highly sensitive and specific, and easy to operate; thereby it could be used for identification of A(H3N2) virus with E119V amino acid change in NA protein.

Amino Acid Substitution ; Drug Resistance, Viral ; Influenza A Virus, H3N2 Subtype ; drug effects ; enzymology ; genetics ; Mutation ; Neuraminidase ; genetics ; Nucleic Acid Probes ; Reverse Transcriptase Polymerase Chain Reaction ; methods

BACKGROUNDWaardenburg syndrome type I (WS1) is an autosomal dominant disorder characterized by sensorineural hearing loss, pigmental abnormalities of the eye, hair and skin, and dystopia canthorum. The gene mainly responsible for WS1 is PAX3 which is involved in melanocytic development and survival. Mutations of PAX3 have been reported in familiar or sporadic patients with WS1 in several populations of the world except Chinese. In order to explore the genetic background of Chinese WS1 patients, a mutation screening of PAX3 gene was carried out in four WS1 pedigrees.

METHODSA questionnaire survey and comprehensive clinical examination were conducted in four Chinese pedigrees of WS1. Genomic DNA from each patient and their family members was extracted and exons of PAX3 were amplified by PCR. PCR fragments were ethanol-purified and sequenced in both directions on an ABI_Prism 3100 DNA sequencer with the BigDye Terminator Cycle Sequencing Ready Reaction Kit. The sequences were obtained and aligned to the wild type sequence of PAX3 with the GeneTool program.

RESULTSTwo nonsense PAX3 mutations have been found in the study population. One is heterozygous for a novel nonsense mutation S209X. The other is heterozygous for a previously reported mutation in European population R223X. Both mutations create stop codons leading to truncation of the PAX3 protein.

CONCLUSIONSThis is the first demonstration of PAX3 mutations in Chinese WS1 patients and one of the few examples of an identical mutation of PAX3 occurred in different populations.

Codon, Nonsense ; Female ; Humans ; Male ; PAX3 Transcription Factor ; Paired Box Transcription Factors ; genetics ; Waardenburg Syndrome ; genetics

To study the prevalence and variation of influenza A(H3N2) viruses, the antigenic and genetic characteristics of influenza A(H3N2) viruses circulating in Mainland China during April 2011 to March 2012 were analyzed. The results showed that influenza A(H3N2) viruses increased gradually since 2012 and became the dominant strain since March. The viruses were antigenically closely related to the vaccine strain A/PER/16/09 (87.2%) and the representative virus A/FJ/196/09 (76.0%) in Mainland China. The genetic characteristics analysis results showed that recently isolated viruses belonged to the Vic/208 clade, and most of the low reaction strains also fell into the same clade. Crystal structure analysis of HA protein found that, compared with the vaccine strain A/PER/16/09, the recently isolated viruses had amino acid substitutions in the antigenic site A, B and C areas, in addition to gaining potential glycosylation sites at the amino acid position of 45 of HA and 367 of NA. Although the majority of circulating influenza A (H3N2) viruses in 2011-2012 season in Mainland China were antigeniclly matched by current influenza vaccine strain and the selected representative viruses, low reaction strains have increased since 2012, therefore it is necessary to strengthen the surveillance on the variation of influenza virus and to provide solid information for the vaccine strain selection.
Amino Acid Sequence ; China ; epidemiology ; Hemagglutinin Glycoproteins, Influenza Virus ; chemistry ; genetics ; Humans ; Influenza A Virus, H3N2 Subtype ; classification ; genetics ; isolation & purification ; physiology ; Influenza, Human ; epidemiology ; virology ; Models, Molecular ; Molecular Sequence Data ; Phylogeny

Pdm09 virus outbreak occurred in Mainland China in May 2009, a few months later, the prevalence of seasonal H1N1(sH1N1) influenza virus that already circulated in human for tens of years began to decline and disappeared afterwards. To identify the reason for the rapid decline of sH1N1 in mainland China, we sequenced the HA1 of sH1N1 during 2006-2011, and then analyzed the selective pressure in different phases. Our results showed before Pdm09 outbreak, the omega value was 0. 36 while after Pdm09 outbreak the omega value was 0. 28 and significant difference (t test, P<0. 05) was identified. We concluded that sH1N1 obtained stronger purifying selection after Pdm09 outbreak in China. This might one of the major reasons causing the disappearance of sH1N1 in human.
China ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; isolation & purification ; Influenza, Human ; virology ; Phylogeny ; Seasons ; Selection, Genetic