OBJECTIVETo construct E1-deletion and replication-defective human type 5 recombinant adenovirus vector and to study the effect of p16INK4a on proliferation and aging of A549 cells.

METHODSp16INK4a cDNA was cloned into pAdCMV to construct recombinant pAdCMV p16INK4a, which was co-transfected into 293 cell together with pJM17. The recombinant p16INK4a adenovirus (Ad-p16INK4a) was generated by homologous recombination and identified with duplex PCR. Lung cancer cell A549, which has a homozygous deletion of p16INK4a gene, was infected with the prepared Ad-p16INK4a virus. X-gal staining and TRAP-ELISA were used for detecting senescence-associated beta-galactosidase and telomerase activities in A549 cells.

RESULTSImmunohistochemical staining and Western blot indicated that p16INK4a gene was transferred into A549 cell with more than 95% efficiency by recombinant adenovirus and p16INK4a protein was expressed at a high level- p16INK4a could markedly inhibit growth of A549 cells, induced expression of senescence-associated beta-galactosidase and suppressed telomerase activity in A549 cells.

CONCLUSIONRecombinant adenovirus vector could efficiently mediate transfer and expression of foreign genes in human cell and could be used for gene immunization and gene therapy; p16INK4a could inhibit A549 cell growth and induce its replicative senescence.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Cellular Senescence ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; physiology ; Gene Expression ; Genetic Vectors ; Humans ; Recombination, Genetic ; Transfection

Objective To study the clinical characteristics and genetic features of SPG4 gene in a family with hereditary spastic paraplegia (HSP). Methods The four patients from one LinYi family were clinically diagnosed as having HSP according to Harding's criteria and their peripheral blood samples were collected. We typed the short tandem repeat (STR) loci closely connected with the known HSP cause gane locus at physical distance and genetic linkage analysis was performed on them. Their haplotypes were structured and then screening of gene mutations was performed. Results Non-elimination of linkage was found between D2S2351 and D2S2255 and cause gene, and the LOD scores in other locus were negative value and eliminated the linkage, which implied that the location was in the ADHSP locus of chromosome 2p22 (SPG4) and the candidate gene was spastin gene. Screening of gene mutations found that the mutation loci lied in heterozygous A and G at nucleotide 1168 in spostin gene. The symptoms of the patients manifested as stiffness, instability or weakness of the legs. Conclusions The patients in this family have typical clinical symptoms of HSP, mainly resulting from the novel mutation (spastin: c1168 A>G).

OBJECTIVETo evaluate the protective effect of ischemic preconditioning against cold ischemia and reperfusion injury of rat intestinal graft following orthotopic transplantation.

METHODSEighty SD rats were randomly assigned into two groups with and without ischemic preconditioning, and each group was divided into 4 subgroups (n=10) according to the intestinal graft cold ischemia time of 3, 6, 12, and 18 h, respectively. Ischemic preconditioning model was established, and the small intestinal graft was preserved at 4 degrees celsius; in Ringer lactate solution for the corresponding time, followed by orthotopic transplantation of the graft. The graft samples were collected for histological examination 1 h after reperfusion, and nuclear factor-kappaB (NF-kappaB) expression in the epithelial cells was detected.

RESULTSIschemia preconditioning obviously relieved the histological ischemia/reperfusion injury, as shown by regular alignment of the small intestinal villi, alleviated muscular layer edema and decreased expression of NF-kappaB in the epithelia of the graft in groups with cold preservation.

CONCLUSIONIschemic preconditioning can protect the intestinal graft from cold ischemia/reperfusion injury, and NF-kappaB is an important cytokine in ischemia preconditioning.

Animals ; Cold Ischemia ; Intestine, Small ; pathology ; transplantation ; Ischemic Preconditioning ; NF-kappa B ; metabolism ; Organ Preservation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control

Objective To explore the effect of long noncoding RNA-ATB (LncRNA-ATB) on phenotypic transition and proliferation of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods HPMCs used in experiment were divided into three groups:control group,mannitol group and hypertonic glucose group.HPMCs in control group received no treatment,and in hypertonic glucose group and mannitol group were treated with 50mmol/L D-glucose and isotonic mannitol for 72 hours,respectively.Real-time PCR was employed to detect the mRNA expression of LncRNA-ATB,E-cadherin,α-smooth muscle actin (α-SMA),connective tissue growth factor (CTGF),Cyclin D1,cyclin dependent kinase inhibitor 4 (CDK4),protein 27 (p27)and proliferating cell nuclear antigen (PCNA).Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA,CTGF,Cyclin D1,CDK4,p27 and PCNA,and flow cytometry was used to test the cell cycle.Lentivirus artifice was used to up-or down-regulate the expression of LncRNA-ATB in untreated HPMCs.Real-time PCR was employed to detect the mRNA expression of E-cadherin,α-SMA and CTGF,Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA and CTGF,and flow cytometry was used to test the cell cycle.Results It is revealed by Real-time PCR,Western blotting and flow cytometry that the expressions increased of LncRNA-ATB,α-SMA,CTGF,Cyclin D1,CDK4 and PCNA induced by hypertonic glucose,and decreased of E-cadherin and p27 (P＜0.05).Up-regulation of LncRNA-ATB promoted HPMCs phenotypic transition and proliferation,while down-regulation alleviated HPMCs phenotypic transition and proliferation.Conclusion Hypertonic glucose may accelerate HPMCs phenotypic transition and proliferation by up-regulating the expression of LncRNA-ATB.

OBJECTIVETo develop a new sampling medium for detecting of bioaerosols.

METHODSThe sampling media were tested by using Escherichia coli, Staphylococcus aureus and Serratia marcescens under static and active conditions, preliminary applications were performed using AGI-10 and high volume sampler.

RESULTSThe average recovery rates were raised to 24.7%, 58.2%, 40.5%, 44.1%, 20.5%, and 15.4%, respectively in six consecutive experiments under static condition for 60 min at room temperature. Four kinds of sampling media were singled out after static experiments, which were referred to as "samplutions" PD1, PX2, TD1, and TX2, respectively. Under the active condition, the protective efficacy of PD1, PX2, TD1, and TX2 was 226% (153/47), 553% (111/17), 150% (120/48), and 268% (419/114), respectively.

CONCLUSIONThe samplutions have some effects on the subsequent nucleic acid detection, which could be avoided by employing standard nucleic acid extraction procedure. The newly developed samplution can be applied to the detection of bioaerosols.

Aerosols ; analysis ; Air Microbiology ; Air Pollutants ; analysis ; Environmental Monitoring ; methods ; Escherichia coli ; isolation & purification ; Nucleic Acids ; isolation & purification ; Sampling Studies ; Serratia marcescens ; isolation & purification ; Staphylococcus aureus ; isolation & purification

Acupuncture Points ; Gastritis, Atrophic ; diagnosis ; therapy ; Health Planning Guidelines ; Humans ; Medicine, Chinese Traditional

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Meningioma ; diagnosis ; Middle Aged ; Retrospective Studies ; Young Adult

BACKGROUNDMeningioma is one of the most common primary tumors of the central nervous system, but there are not many detailed studies on the sex, age, subtypes and locations of large series. This study was a retrospective analysis of the characteristics of meningioma cases consecutively operated on at a single institution in China from 2001 to 2010.

METHODSThis study investigated the demographic background of 7084 meningioma cases, and the subtypes and locations of the tumors. Sex and age distributions were analyzed, and the pathological subtypes were classified according to the World Health Organization (WHO) classification. The location of the meningiomas was also categorized.

RESULTSThe female:male ratio of the 7084 cases was 2.34:1. The mean age was 51.4 years (range, 11 months-86 years). The mean age of cases of WHO grade I meningioma was significantly older than that of grade II or III meningiomas (P < 0.001, Fisher's Least Significant Digit test). There was a significantly higher female:male ratio in WHO grade I meningiomas than in grade II or grade III meningiomas (2.57, 1.03 and 0.76, respectively; P < 0.001, χ(2) test). Meningothelial (n = 2061) and fibrous meningiomas (n = 3556) were the most common subtypes, comprising 79.3% of all meningiomas. All meningioma cases were classified into 23 locations in this study, with the cerebral convexity the most common site (38.33%, n = 2722). Cases with uncommon locations such as extra-cranial and sylvian fissure meningiomas were also present in this series.

CONCLUSIONSFemale predominance was found for benign meningiomas, while malignant subtypes showed male predominance. The mean age of patients with WHO grade I meningiomas was older than that of patients with higher-grade tumors. Meningothelial and fibrous meningiomas were the most common subtypes. The cerebral convexity was the most common meningioma location.

Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Meningioma ; epidemiology ; Middle Aged ; Sex Distribution ; Young Adult

OBJECTIVE: To investigate the mechanism of Tojapride, a Chinese herbal formula extract, on strengthening the barrier function of esophageal epithelium in rats with reflux esophagitis (RE). METHODS: Ten out of 85 SD rats were randomly selected as the sham group (n10), and 75 rats were developed a reflux esophagitis model (RE) by the esophageal and duodenal side-to-side anastomosis. Fifty successful modeling rats were divided into different medicated groups through a random number table including the model, low-, medium-, and high-dose of Tojapride as well as omeprazole groups (n10). Three doses of Tojapride [5.73, 11.46, 22.92 g/(kg•d)] and omeprazole [4.17 mg/(kg•d)] were administrated intragastrically twice daily for 3 weeks. And the rats in the sham and model groups were administered 10 mL/kg distilled water. Gastric fluid was collected and the supernatant was kept to measure for volume, pH value and acidity. Esophageal tissues were isolated to monitor the morphological changes through hematoxylin-eosin (HE) staining, and esophageal epithelial ultrastructure was observed by transmission electron microscopy. The expressions of nuclear factor kappa-light-chain-enhancer of activated B cells p65 (NF-KBp65), κB kinase beta (IKKß), occludin, and zonula occludens-1 (ZO-1) in the esophageal tissues were measured by immunohistochemistry and Western blot, respectively. RESULTS: The gastric pH value in the model group was significantly lower than the sham group (P<0.05). Compared with the model group, gastric pH value in the omeprazole and medium-dose of Tojapride groups were significantly higher (P<0.05). A large area of ulceration was found on the esophageal mucosa from the model rats, while varying degrees of congestion and partially visible erosion was observed in the remaining groups. Remarkable increase in cell gap width and decrease in desmosome count was seen in RE rats and the effect was reversed by Tojapride treatment. Compared with the sham group, the IKKß levels were significantly higher in the model group (P<0.05). However, the IKKß levels were down-regulated after treatment by all doses of Tojapride (P<0.01 or P<0.05). The occluding and ZO-1 levels decreased in the model group compared with the sham group (Ps0.01 or Ps0.05), while both indices were significantly up-regulated in the Tojapride-treated groups (P<0.01 or P<0.05). CONCLUSIONS Tojapride could improve the pathological conditions of esophageal epithelium in RE rats. The underlying mechanisms may involve in down-regulating the IKKß expression and elevating ZO-1 and occludin expression, thereby alleviating the inflammation of the esophagus and strengthening the barrier function of the esophageal epithelium.