OBJECTIVETo construct a lentiviral-vector-mediated CyPA small interference RNA (siRNA) and study its function in non-small cell lung cancer.

METHODSFirst, four target sequences were selected according to CyPA mRNA sequence, the complementary DNA contained both sense and antisense oligonucleotides were designed, synthesized and cloned into the pGCL-GFP vector, which contained U6 promoter and green fluorescent protein (GFP). The resulting lentiviral vector containing CyPA shRNA was named Lv-shCyPA, and it was confirmed by PCR and sequencing. Next, it was cotransfected by Lipofectamine 2000 along with pHelper1.0 and pHelper 2.0 into 293T cells to package lentivirus particles. At the same time, the packed virus infected non-small cell lung cancer cell (A549), the level of CyPA protein at 5 d after infection was detected by Western Blot to screen the target of CyPA. A549 were infected with Lv-shCyPA and grown as xenografts in severe combined immunodeficient mice. Cell cycle and apoptosis were measured by FCM.

RESULTSIt was confirmed by PCR and DNA sequencing that lentiviral-vector-mediated CyPA siRNA (Lv-shCyPA) producing CyPA shRNA was constructed successfully. The titer of concentrated virus were 1 x 10(7) TU/ml. Flow cytometric analysis demonstrated G2-M phase (11.40% +/- 0.68%) was decreased relatively in A549/LvshCyPA compared with control groups (14.52% +/- 1.19%) (P<0.05). The apoptosis rate of A549/Lv-shCyPA (5.01% +/- 0.5%) was higher than control groups (0.35% +/- 0.17%) (P<0.05). Visible tumors were only detectable at 6th day after inoculated by A549/Lv-shCyPA. The xenograft tumors of A549/Lv-shCyPA remarkably delayed tumor growth and remained at a similarly small average size at 38th days after inoculation compared with the control group (P < 0.05).

CONCLUSIONLentiviral-vector-mediated siRNA technique effectively inhibits the expression of CyPA, induces the NSCLC cell apoptosis, inhibits the tumor growth. Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for non-small cell lung cancer.

Animals ; Carcinoma, Non-Small-Cell Lung ; genetics ; Cell Line, Tumor ; Cyclophilin A ; genetics ; Gene Silencing ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Lung Neoplasms ; genetics ; Mice ; RNA, Small Interfering

OBJECTIVETo prevent chronic severe hepatitis, even more fulminant hepatic failure (FHF) occurrence in patients with chronic hepatitis B of severe degree using steroid.

METHODS120 patients were randomized into conventional supporting treatment and steroid treatment groups. The latter, 62 patients were given intravenously hydrocortisone sodium succinate at the dose of 150 mg to approximately 200 mg everyday plus support care.

RESULTSThe rate of deteriorating to chronic severe hepatitis in steroid treatment group was significantly lower than that of conventional group (22% vs 48%, x(2) =7.60, P<0.01). 53.6% (15/28) patients with chronic severe hepatitis in conventional group died, while only 28.6% (4/14) in steroid treatment group succumbed to terminal liver disease (x(2)=0.02, P>0.05). There was no difference between the two groups regarding to complications incidence: gastrointestinal bleeding and infections except for some controllable serious reverse events, such as candidiasis, diabetes, herpes zoster and pulmonary tuberculosis found in some patients in steroid-treated group.

CONCLUSIONThese results suggest that steroid administration with improved support care not only is likely to prevent chronic severe hepatitis occurrence in patients with chronic viral hepatitis of severe degree, but also shows some efficacy for FHF, which warrant further investigation.

Adult ; Female ; Glucocorticoids ; therapeutic use ; Hepatitis B, Chronic ; complications ; drug therapy ; Humans ; Liver Failure ; prevention & control ; Male

OBJECTIVETo discuss CT and MRI characteristics of hepatic angiomyolipoma based on pathological findings.

METHODSThe CT and MRI appearances with related pathohistological subtypes of 11 hepatic angiomyolipomas from 10 patients were retrospectively analyzed.

RESULTTen patients with hepatic angiomyolipomas were subcategorized into lipomatous (3 cases), angiomatous (1 case), myomatous (1 case) and mixed (5 cases) subtypes. Lesions of the lipomatous type were mainly composed of adipocytes which could be easily recognized on both CT and MRI. Abnormal vessels were commonly seen in the angiomatous lesions, which showed pronounced enhancement in the early arterial phase and remained higher than or isodense with the normal parenchyma in the portal phase. The myomatous type was predominantly composed of leiomyoid cells mixed with small amount of adipocytes. The mixed type was the most frequent,evenly comprising sheets of epithelioid muscle cells admixed with islands of adipocytes and abnormal vessels, and showing homogeneously low density on plain CT and low signal intensity on T1-weighted,intermediately high signal intensity on T2-weighted MRI scans. On dynamic study with both CT and MRI, the mixed type exhibited obvious enhancement, which retained to some degree during the portal phase.

CONCLUSIONThe discrete CT and MRI appearances of hepatic angiomyolipomas with different pathological subtypes depend on the components of the tumor.

Adult ; Angiomyolipoma ; pathology ; Female ; Humans ; Liver Neoplasms ; blood supply ; pathology ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Neovascularization, Pathologic ; diagnostic imaging ; Retrospective Studies ; Tomography, X-Ray Computed ; methods

OBJECTIVETo investigate the effect of periodontal infection on circulating C-reactive protein (CRP) in type 2 diabetes patients.

METHODS32 diabetes patients with advanced periodontitis participated in this study. They were compared to a group of 32 diabetes patients without periodontal disease, who were mathed with regard to age (+/- 3 years), gender and body mass index (+/- 1 kg/m2). The concentration of CRP on circulation was measured by ELISA.

RESULTSSignificant difference was found in the level of CRP and the percentage of subjects with elevated CRP levels > or = 3 mg/L on circulation between the two groups(P < 0.05).

CONCLUSIONPeriodontal infection results in higher circulating CRP in type 2 diabetes patients. This elevated inflammatory factor may exacerbate insulin resistance and increase the risk for great vessels complications of diabetes mellitus.

Adult ; C-Reactive Protein ; Diabetes Mellitus, Type 2 ; Female ; Humans ; Male ; Middle Aged ; Periodontal Attachment Loss ; Periodontal Pocket ; Periodontitis

OBJECTIVEThe aim of this study is to investigate the changes of hard tissue profile in anterior-posterior and vertical direction in bimaxillary protrusion patients after orthodontic treatment.

METHODSA total of 24 bimaxillary protrusion patients (male 8, female 16), aged from 11.2 to 29.0 (average 16.9 years old), were selected to be treated with standard edgewise technique consisted of 4 first premolars extraction. Cephalometrics were taken before and after treatment. The changes of hard tissue profile were studied using the computer-aid X-ray cephalometric analysis.

RESULTS1. The length of the maxillary and the mandible increased significantly, but the anterior-posterior relationship of the maxillary and the mandible did not change significantly. 2. The anterior and posterior facial height increased significantly, but the ratio of anterior and posterior facial height and the angle of MP-FH which reflected the inclination of the mandible plane did not change significantly. 3. The height of the upper and lower first molar increased significantly along with the increase of the anterior and posterior facial height. 4. The height of the upper incisors increased significantly, but the height of the lower incisors decreased significantly.

CONCLUSIONThe anchorage in anterior-posterior and vertical direction were controlled preferably, the patients did not manifest disadvantageous vertical growth trend.

Adolescent ; Adult ; Cephalometry ; Child ; Facial Bones ; pathology ; Female ; Humans ; Male ; Orthodontic Wires ; Orthodontics, Corrective ; Prognathism ; therapy ; Skull ; pathology

OBJECTIVETo summarize clinical and molecular features of two children with autosomal recessive chronic granulomatous disease caused by CYBA mutations.

METHODThe clinical records and CYBA mutations were reviewed for analysis of infections and inflammatory complications.

RESULTThe first case was a girl diagnosed with "liver and spleen abscess" in our hospital when she was 2.9 years old, with past history of neonatal impetigo and recurrent purulent lymphadenitis and positive family history. The results of DHR123 flow-cytometry showed that positive phagocytes after phorbol ester (PMA) stimulation was 84.63%. CYBA mutation analysis showed that she had heterozygous 35C > T, Q3X and IVS-2A > G. The second case was a boy diagnosed with "sepsis (salmonella D)" when he was 4 years old with a past history of impetigo, sepsis, perianal abscess, skin infection and positive family history. The results of flow cytometry showed that positive phagocytes after PMA stimulation was 96.13%. CYBA mutation analysis showed that he had homozygous 35C > T, Q3X and his parents were all carriers. All of them had BCG related axillary lymphnode calcification.

CONCLUSIONA22CGD cases had recurrent purulent infections (skin, lymphnode, liver and spleen, lung, blood), DHR123 flow cytometric analysis helped the diagnosis of CGD, CYBA mutation analysis ascertained the diagnosis of A22CGD.

Child, Preschool ; Chromosome Aberrations ; DNA Mutational Analysis ; Female ; Genes, Recessive ; Granulomatous Disease, Chronic ; diagnosis ; genetics ; Homozygote ; Humans ; Male ; Mutation ; NADPH Oxidases ; genetics

OBJECTIVETo investigate the correlation between intercellular adhesion molecule1 (ICAM1) gene K469E polymorphism and coronary heart disease(CHD) in Han Chinese population.

METHODSUsing the methods of polymerase chain reaction-restrictive fragment length polymorphism (PCR-RFLP), 173 CHD patients and 141 controls were analyzed for the polymorphism, genotype and allele distribution of ICAM1 gene K469E.

RESULTSThe distribution of ICAM1 genotypes was in Hardy-Weinberg equilibrium. The frequency of KK genotype in CHD group was significantly higher than that in control (64.2% vs 48.9%, P<0.01). Similarly, the frequency of K allele in CHD group was significantly higher than that in control (79.2% vs 69.9%, P<0.01). With Logistic Regression Analysis ruling out the influences of age, gender and other CHD risk factor, the homozygous individual with KK genotype was 2.35 folds of KE or EE genotype one suffering from CHD (OR: 2.35, 95%CI: 1.03-5.36, P<0.05).

CONCLUSIONICAM1 gene K469E polymorphism is associated with CHD risk of Han Chinese population, the K allele may serve as a genetic risk factor of coronary heart disease.

Amino Acid Substitution ; Asian Continental Ancestry Group ; genetics ; Coronary Disease ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Logistic Models ; Male ; Polymorphism, Genetic ; Risk Factors

Objective    To discuss the clinical characteristics and the management of major complications after thoracic surgery. Methods    Retrospective research was conducted on 15 213 patients who underwent thoracic surgery from January 2008 to September 2018 in our hospital. Thirty-six (0.24%) patients died of postoperative complications. Based on whether major complications such as severe pulmonary pneumonia and other 13 complications were presented postoperatively, the patients were divided into a complication group (n=389, 294 males and 95 females, aged 61.93±10.23 years) and a non-complication group (n=14 785, 8 636 males and 6 149 females, aged 55.27±13.21 years) after exclusion of unqualified patients. The age, gender distribution, diagnosis, surgical approach, postoperative hospital stay, in-hospital costs and other clinical data were analyzed. And the treatment and outcomes of the complications were summarized. Results    The age, proportion of male, malignancy and esophageal diseases, postoperative hospital stay and in-hospital costs in the complication group were significantly more or higher than those in the non-complication group (P<0.05). The top three causes of death among the 36 deaths were pulmonary embolism (PE, 25.00%), severe pulmonary pneumonia (16.67%) and acute respiratory failure (16.67%), respectively. The top five complications among the severe complication group were pulmonary pneumonia (24.73%), pleural space (19.83%), anastomotic leak (17.48%), pulmonary atelectasis (11.51%) and PE (6.18%). Conclusion    Thoracic surgeons should recognize patients with high risk of severe complications preoperatively based on clinical characteristics and perform multi-disciplinary treatment for severe complications.

OBJECTIVETo construct a replication-incompetent recombinant adenovirus mediating short hairpin RNA (shRNA)-induced tissue factor gene silencing in the islet.

METHODSFour pairs of complementary oligonucleotides were designed and synthesized to create double-stranded oligonucleotides (ds oligo). The ds oligos were cloned into Pentr/U6 vector to construct the shuttle plasmid pENTR/U6-shRNA, which was transduced into human islets via liposome after sequence verification. The plasmid with the best silencing effect was identified by real-time RT-PCR, followed by homologous recombination with the adenovirus backbone plasmid. The functional clone was transfected into 293A cells to amplify the adenovirus, whose silencing effect against TF expression was tested using real-time RT-PCR and Western blotting.

RESULTSThe pENTR/U6-shRNA shuttle plasmid was constructed and verified by sequencing. The recombinant adenovirus-mediated shRNA against TF was constructed, and real-time RT-PCR and Western blotting demonstrated that the strongest silencing effect of the adenovirus against TF occurred on the 4th day following islet transfection.

CONCLUSIONReplication-incompetent recombinant adenovirus-mediated shRNA against TF has been successfully constructed, which has good silencing effect against TF expression in human islet in vitro.

Adenoviridae ; genetics ; physiology ; Base Sequence ; Cell Line ; DNA, Recombinant ; genetics ; Gene Expression ; Genetic Engineering ; methods ; Humans ; Inverted Repeat Sequences ; Islets of Langerhans ; metabolism ; Plasmids ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Thromboplastin ; deficiency ; genetics ; Viral Load ; Virus Replication

OBJECTIVETo investigate the availability of serum level of macrophage clony stimulating factor (M-CSF) as a marker for early diagnosis of Alzheimer's disease (AD).

METHODSThe serum levels of M-CSF in 70 patients with AD, 52 healthy controls, 22 patients with VAD (vascular dementia) were measured and the serum levels of IL-1 beta, IL-6, TNF-alpha in 32 patients with AD and 20 controls were measured as well.

RESULTSSerum levels of M-CSF were significantly elevated in patients with AD when compared with healthy controls (P < 0.01) and VAD controls (P < 0.05) respectively. At the early stage of mild dementia and middle dementia, serum levels of M-CSF were significantly elevated, but at the later stage of severe dementia, they returned to normal level. Serum levels of IL-1 beta were significantly elevated in AD patients compared with controls (P < 0.05), and serum levels of TNF-alpha and IL-6 were within the normal range in patients with AD.

CONCLUSIONSThe results suggest that serum M-CSF level may provide a convenient and sensitive means for the early diagnosis of Alzheimer's disease.

Aged ; Aged, 80 and over ; Alzheimer Disease ; blood ; Biomarkers ; blood ; Female ; Humans ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Macrophage Colony-Stimulating Factor ; blood ; Male ; Middle Aged ; Tumor Necrosis Factor-alpha ; metabolism