AIM:Targeting of focal adhesion kinase (FAK) gene,we aim to construct FAK shRNA lentiviral vector and to identify its function on the growth of leukemic cells.METHODS:FAK shRNA was chemically synthesized,and inserted into a GFP-lentiviral plasmid through molecular biology methods.After packaged and concentrated,the lentiviral-FAK-shRNA-vector was transduced into a human leukemic cell line.FAK gene expression was detected by reverse transcriptional PCR and Western blotting.Cell apoptosis was measured by Annexin V labeling.RESULTS:The results showed that FAK shRNA was successfully inserted into the lentival vector,and the infection efficiency varied from 10% to 25%.Compared to the control vector (lentival vector without FAK shRNA),FAK shRNA inhibited the expression of FAK mRNA and protein by 40% and 70%,respectively.Moreover,the results of apoptosis experiment showed that the percentages of Annexin V+ cells in control vector group and FAK shRNA group were (4.19 ? 0.36) % and (8.48 ? 0.58) % respectively,the difference was statistically significant (P

BackgroundWith the increasing prevalence of dry eye and the continuous improvement of living standards,the problem of dry eye more and more get the attention of people.At present,China still lacks the large population-based epidemiological data of dry eye. Objective To investigate the prevalence and possible risk factors of dry eye in a community of Huidong County of population aged 14 and over.Methods From September 2010 to January 2011,using questionnaires and examination of dry eye related,2800 people were selected randomly for cross-sectional survey.Those suspected as dry eye were examed by the SchirmerⅠtest ( S Ⅱ T),tear-film breakup time(BUT),corneal fluorescein staining(F1).Results In the 2475 questionnaire effectively,154 persons were diagnosed as dry eye,and the prevalence rate of dry eye was 6.22％,8.06％in females,4.14％in males.The prevalence rate increases with age.The S Ⅰ T and BUT decreased with increasing age.S Ⅰ T and BUT in females are less than males.Foreign body sensation is the primary complaints of patients.Logistic analysis showed that the most common risk factors in dry eye are age and gender.The system disease and eye diseases,eye fatigue and long exposure to dust are also main determinants.ConclusionsThe population prevalence rate of dry eye increased with age,the prevalence rate of dry eye in females is higher than that in males.The key factors associated with dry eye are age,gender,systemic disease and eye diseases,occupation,working environment.

Objective To investigate the inhibitory effect and mechanism of soybean-derived Bowman-Birk inhibi-tor (BBI)on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells.Methods Cytotoxicity effect of LPS and BBI on intestinal epithelial cells was analyzed by MTT assay.Intestinal epithelial cells were pre-treated with BBI,followed by LPS stimulation,expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1)was detected by quantitative real-time polymerase chain reaction;the activation of NF-κB was measured by pNF-κB-luc system and Western Blot.Results The maximum concentration of LPS (10000 ng/mL)and BBI (1000 μg/mL)had no cytotoxicity effect on intestinal epithelial cells.LPS could potently up-regulate the expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1 ),the up-regulation was positively correlated to the concentration of LPS;LPS-induced expression of inflammatory cytokines in intestinal epithelial cells could achieve the highest level,then decreased over time.The up-regulation of LPS on inflammatory cytokines in intestinal epi-thelial cells had dose-time effect;when intestinal epithelial cells were pretreated by BBI for 6 hours,the inhibitory effect of BBI on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells was most obvious, and had dose-time effect.Conclusion BBI can potently inhibit LPS-induced expression of inflammatory cytokines through inhibiting NF-κB in intestinal epithelial cells.

OBJECTIVETo observe the therapeutic angiogenesis effect of naotai recipe (NR) on local ischemia/reperfusion (I/R) injury of rats by observing signaling pathway of hypoxia-inducible factor-lα (HIF-1α) and vascular endothelial growth factor (VEGF).

METHODSTotally 120 Sprague-Dawley (SD) rats were randomly divided into 4 groups, namely, the normal control group (n =12), the sham-operation group (n =12), the I/R model group (n =48), and the NR group (n =48). Cerebral I/R injury models were established using thread suture method. Rats in the I/R model group and the NR group were sub-divided into 4 sub-groups according to the 1st, 3rd, 5th, and 7th I/R day (n =12). The phenomenon of neovasculization was observed by immunofluorescence staining. The protein and mRNA expression levels of HIF-la, VEGF-A, and VEGFR II receptor were detected by RT-PCR.

RESULTSThere were a large amount of labels for neovasculization in the ischemic area of the NR group. Double-immunofluorescence labeling [vWF (red) and BrdU (green)] was observed in the NR group. Compared with the model group, the HIF-1α protein expression was obviously enhanced on the 1 st day of I/R (P <0.01), and the VEGF protein expression started to enhance on the 3rd day in the NR group (P <0.01). The VEGFR protein expression level was the highest in the NR group on the 5th day of I/R (P <0.01). The protein expression of VEGF and HIF-1α started to decrease on the 7th day of I/R.

CONCLUSIONNR could strengthen angiogenesis after I/R by elevating the expression of HIF-lα and activating HIF-lα/VEGF signaling pathway.

Animals ; Brain Ischemia ; metabolism ; Cerebral Infarction ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Hypoxia-Ischemia, Brain ; metabolism ; Ischemia ; Neovascularization, Pathologic ; Rats, Sprague-Dawley ; Reperfusion Injury ; Signal Transduction ; Vascular Endothelial Growth Factor A ; biosynthesis

BACKGROUNDOur previous papers indicate that flurbiprofen axetil (FA), a cyclooxygenase inhibitor, is a promising therapeutic strategy for cerebral ischemia in rats. This study aimed to investigate whether FA could promote a neuroprotective effect by activation of peroxisome proliferator-activated receptor-γ (PPAR-γ) after focal cerebral ischemia in rats.

METHODSTotally 48 male Sprague-Dawley (SD) rats were randomly assigned into six groups (n = 8 in each group): animals in group ischemia/reperfusion (I/R) only received 120-minute transient middle cerebral artery occlusion (tMCAO); animals in group I/R + FA were administered FA (10 mg/kg) by caudal vein just after 120-minute tMCAO; animals in group I/R + FA + GW9662 were administered GW9662 (a PPAR-γ inhibitor, 1 mg/kg) intraperitoneally 30 minutes before cerebral ischemia onset and FA (10 mg/kg) by caudal vein just after 120-minute tMCAO; animals in group I/R + GW9662 were administered GW9662 (1 mg/kg) intraperitoneally 30 minutes before cerebral ischemia onset; animals in group I/R + DMSO were administered 3% DMSO (vehicle of GW9662, 1 ml/kg) intraperitoneally 30 minutes before cerebral ischemia onset; animals in sham group experienced the identical surgery apart from the insertion of the nylon filament. The neurologic deficit score (NDS) were performed at 72 hours after reperfusion, and then mean brain infarct volume percentage (MBIVP) was determined with 2,3,5-triphenyltetrazolium chloride (TTC) 10 g/L staining.

RESULTSNDS was significantly increased in group I/R + FA (12.0 (10.0 - 15.0)), group I/R + FA + GW9662 (10.0 (8.0 - 12.0)), and in group I/R + FA + DMSO (12.0 (9.0 - 14.0)) at 72 hours after reperfusion compared with those in group I/R (7.5 (6.0 - 10.0)). NDS was conspicuously different between group I/R + FA (12.0 (10.0 - 15.0)) and group I/R + FA + GW9662 (10.0 (8.0 - 12.0)). MBIVP in group I/R ((45.82 - 8.83)%) was significantly greater than that in group I/R + FA ((23.52 - 9.90)%), group I/R + FA + GW9662 ((33.17 - 7.15)%); MBIVP in group I/R + FA ((23.52 - 9.90)%) was significantly smaller than that in group I/R + FA + GW9662 ((33.17 - 7.15)%).

CONCLUSIONSFA confers the neuroprotective effect on tMCAO in rats and the selective PPAR-γ antagonist GW9662 attenuates the effect of FA. FA could promote a neuroprotective effect by, or in part, activation of PPAR-γ after focal cerebral ischemia in rats.

Animals ; Brain Ischemia ; drug therapy ; Cyclooxygenase Inhibitors ; pharmacology ; Flurbiprofen ; analogs & derivatives ; pharmacology ; Male ; Neuroprotective Agents ; pharmacology ; PPAR gamma ; physiology ; Rats ; Rats, Sprague-Dawley

Important forensic diagnostic indicators of sudden death in coronary atherosclerotic heart dis-ease,such as acute or chronic myocardial ischemic changes,sometimes make it difficult to locate the ischemic site due to the short death process,the lack of tissue reaction time.In some cases,the de-ceased died of sudden death on the first-episode,resulting in difficulty for medical examiners to make an accurate diagnosis.However,clinical studies on coronary instability plaque revealed the key role of coronary spasm and thrombosis caused by their lesions in sudden coronary death process.This paper mainly summarizes the pathological characteristics of unstable coronary plaque based on clinical medi-cal research,including plaque rupture,plaque erosion and calcified nodules,as well as the influencing factors leading to plaque instability,and briefly describes the research progress and technique of the atherosclerotic plaques,in order to improve the study on the mechanism of sudden coronary death and improve the accuracy of the forensic diagnosis of sudden coronary death by diagnosing different patho-logic states of coronary atherosclerotic plaques.

OBJECTIVETo investigate the presence of abnormal metabolism in the thalamus and hypothalamus in patients with first-episode depression.

METHODSThirty drug-naive patients with first-episode depression and 30 age-matched controls were scanned with proton magnetic resonance spectroscopy ((1)H-MRS) for Naa, Cho, Cr and mI.

RESULTSCompared with the control group, the patients showed significantly reduced mI and mI/Cr of the hypothalamus, reduced mI/Cr of the left thalamus, and lowered Cho, ml, and ml/Cr of the right thalamus (P<0.05).

CONCLUSIONPatients with first-episode depression may have myo-inositol and phosphoric acid metabolism disorder in the thalamus and hypothalamus with malfunction of cellular osmotic pressure adjustment mechanism. Abnormal mI/Cr in the thalamus and hypothalamus may represent an important biochemical change in advanced patients with depression.

Adolescent ; Adult ; Case-Control Studies ; Choline ; metabolism ; Creatine ; metabolism ; Depression ; diagnosis ; Female ; Humans ; Hypothalamus ; metabolism ; Inositol ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Male ; Middle Aged ; Protons ; Thalamus ; metabolism ; Young Adult

BACKGROUNDTo understand the antigenic and genetic characteristics of Victoria like strain of influenza B virus isolated recently and to provide a scientific evidence for influenza surveillance and monitoring of influenza epidemic in future.

METHODSViruses were passed in embryonated hen eggs and virion RNA was extracted from allantoic fluid and reverse transcribed to synthesize cDNA. cDNA was amplified by PCR and the PCR product was purified with a purification kit. Afterwards RNA sequence analysis was performed by dideoxynucleotide chain termination and a cloning method. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign.

RESULTSB/Sichuan/63/2001 and B/Zhejiang/2/2001 viruses were antigenically different from B/Shandong/7/97 strain. The substitution of nucleotide sequences of HA1genes of them compared with those of B/Shandong/7/97strain resulted in the change of amino acid sequences in antigenic determinants on HA1 protein domain. The phylogenetic analysis also indicated that strains isolated recently were genetically different from B/Shandong/7/97/strain. However, there was neither differences on the antigenicity nor genetic partern between these two isotates.

CONCLUSIONSThe antigenic drift of Victoria-like strain of influenza B virus isolated recently in China has further occurred.

Amino Acid Sequence ; Antigenic Variation ; China ; Epitopes ; Genetic Variation ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; immunology ; Humans ; Influenza B virus ; genetics ; immunology ; Influenza, Human ; virology ; Molecular Sequence Data ; RNA, Viral ; analysis ; Sequence Analysis, RNA

OBJECTIVETo summarize the clinical characteristic and outcome of digital gigantism of the foot.

METHODSRetrospectively analyze the clinical documents of cases of digital gigantism of the foot. Twelve 12 cases with 13 feet in this study included 8 male and 4 female with an average 4.6-years-old. All the deformities were found at birth. Multiple toes involved were more than single toe, and tibial toe involved more than fibular. Forefoot was enlarged. All the phalanges involved and partial metatarsal bones were enlarged. Marked increase in subcutaneous fat was found in all cases in the operation which infiltrated interossei and articular capsules. The appearance of the nerves and its branches in the foot were normal and fat infiltrating was not discovered. The operation types included debulking, epiphyseal arrest, amputation, nerve stripping and anastomosis.

RESULTSSeven cases were followed up with mean periods 25.6 months. Functional evaluation according to a criterion formulated by author revealed a result of 2 excellent, 2 good and 3 fair.

CONCLUSIONSDigital gigantism of the foot is an uncommon congenital deformity of the foot characterized by overgrowth of both the soft-tissue and the osseous elements of the enlarged toe and forefoot. Surgical treatment is the unique method, and the goal is to reduce the size of the foot to allow fitting regular shoes and walking readily. There are several types of operations which to be chosen. The indication, the timing of operative intervention and the selection of operation type should be paid more attention.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Foot Deformities, Congenital ; surgery ; Forefoot, Human ; surgery ; Humans ; Infant ; Male ; Retrospective Studies ; Toes ; abnormalities ; Treatment Outcome

OBJECTIVETo understand the epidemics and antigenic drift of influenza viruses in China from 2000 to 2001.

METHODSThe viruses were grown in embryonated hen eggs with 9 - 10 days old. The egg allantoic fluids with influenza viruses were used. Virion RNA was transcribed into cDNA by reverse transcriptase while cDNA amplified by PCR. Products of PCR were purified. RNA sequence analysis was then performed. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (Version 1.03) and Editseq (Version 3.69) software.

RESULTSData from comparison of amino acid sequence on HA1 domain of HA protein molecule between H1N1 viruses isolated in 2001 and A/Shanghai/7/99 (H1N1) strain indicated that there was only one difference of amino acid located at 190 position (antigenic determinant D). However, phylogenetic analysis showed that there were two distinguishable genetically lineages of H1N1 viruses co-circulating in men in China in 2001. Two antigenically distinct genetic lineages of influenza B viruses were still existing in men in China. Most of influenza B viruses were Yamagata-like strain and there were two different amino acid sequences located at 197 and 199 position on HA1 domain of HA protein molecule, between Victoria-like virus isolated and B/Shandong/7/97 strain. When comparing amino acid sequences on HA1 protein domain of H3N2 viruses isolated in 2000 with those of A/Sydeney/5/97 (H3N2) virus, it was revealed that there were 7 - 8 differences of amino acid sequences between them. However, there were four differences related to amino acid sequences on HA1 protein domain between H3N2 viruses isolated in 2000 and in 2001. These results were further demonstrated by analysis of phylogenic tree.

CONCLUSIONSInfluenza was not prevalent in China from 2000 to 2001. The antigenic drifts of H3N2 and B/Victoria-like viruses occurred. Two antigenically distinct genetic lineages of influenza B viruses were still co-circulating in men in China. Two genetically distinct lineages of influenza A (H1N1) virus were also co-circulating in men in China.

Antigens, Viral ; genetics ; China ; epidemiology ; DNA, Viral ; genetics ; Female ; Genes, Viral ; genetics ; Genetic Variation ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A virus ; classification ; genetics ; immunology ; isolation & purification ; Influenza B virus ; classification ; genetics ; immunology ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Male ; Orthomyxoviridae ; classification ; genetics ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sentinel Surveillance ; Sequence Analysis, RNA