Objective To construct a luciferase reporter plasmid containing interferon regulatory factor 3(IRF-3)human gene promoter and to evaluate promoter activity in human embryonic kidney(HEK)-293 cells.Methods The 1 000 bp fragment was amplified by PCR with human genomic DNA as a template and was directionally cloned into pGL3-basic multiple cloning sites to construct the luciferase repor-ter plasmid pGL3-pIRF-3.Transfection of HEK-293 cells with the promoter-driven lucife-rase construct was performed to induce lucife-rase gene expression and calculate the relative luciferase activity unit(RLU).Promoter sequence of 1 000 bp upstream of transcription initiation site of IRF-3 was analyzed by using Promoter 2.0 Prediction software.Results DNA sequencing and restriction endonuclease analysis verified the successful construction of the plasmid pGL3-pIRF-3.This IRF-3 promoter exhibited a strong promoter activity with an increase of 42.2-fold of RLU in HEK-293 cells when compared with pGL-3 basic vector.The transfection experiment confirmed that the levels of its activation were significantly higher than that in controls in HEK-293 cells.Function analysis of IRF-3 promoter disclosed seve-ral GATA-1 and specific protein 1(Sp1) sites and E2F in minimal promoter region.Conclusion The plasmid pGL3-pIRF-3 promoter is successfully constructed and has a strong basal promoter activity in HEK-293 cells.

Acetylcysteine ; analogs & derivatives ; urine ; Adolescent ; Adult ; Deoxyguanosine ; analogs & derivatives ; urine ; Female ; Furans ; urine ; Humans ; Hydrocarbons, Aromatic ; analysis ; Male ; Occupational Exposure ; analysis ; Young Adult

OBJECTIVE:To optimize the formulation of Budesonide sustained-release tablet. METHODS:Using the cumula-tive releases in 2,4,8 h as investigation indexes,central composite design-response surface method was used to optimize the amount of hydroxypropylcellulose L(HPC-L),amount of soybean phosphatides,and filler(fixed total 200 mg)lactose- micro-crystalline cellulose mass ratio in the formulation of Budesonide sustained-release tablet,and the verification test was conducted. The release behaviors of prepared sustained-release tablet and original preparation in pH 7.2,7.0,6.8 phosphate buffer were com-pared. RESULTS:The optimal formulation was as follow as budesonide of 9 mg,HPC-L of 46.49 mg,soybean phosphatides of 9.23 mg,filler lactose-microcrystalline cellulose mass ratio of 1:2.9;the cumulative releases in 2,4,8 h were 21.9%,50.1%, 99.5%,the relative errors with predicted values (22.0%,50.0%,98.5%) were 0.45%,0.20%,1.02%(n=3),respectively. Compared with cumulative release of original preparation,the f2 was higher than 50. CONCLUSIONS:Budesonide sustained-re-lease tablet is successfully prepared,which shows similar release behavior to original preparations.

Objective To analyze the Alb-cre/DTR mouse phenotype, and establish a model of induced liver damage to serve basic researches of liver diseases.Methods The introduced Alb-cre and DTR mice were crossed to obtain Alb-cre/DTR mice and the genomic DNAs were extracted from the tail tissue of the mice for genotying by PCR.Diphtheria toxin was intraperitoneally(i.p.)injected into the Alb-cre/DTR mice, then the body weights were monitored and the sera were collected for the detection of serum ALT and AST levels.Results By crossing Alb-cre and DTR mice we obtained the Alb-cre and DTR double transgenic mouse.The intraperitoneal injection of diphtheria toxin in a dose of 0.625 ng/g body weight significantly induced liver injury in these mice, as showed by the elevated levels of ALT and AST, the gross appearance of liver damage and the pathological changes such as necrosis in the liver tissue.Conclusions We have ob-tained a novel mouse strain of Alb-cre/DTR by crossing Alb-cre and DTR mice.Liver damages in those Alb-cre/DTR mice can be induced by injection of diphtheria toxin.This established mouse model of inducible liver damage is a useful platform for the studies of liver damage and recovery, as well as liver transplantation.

Objective To investigate the role of interleukin-1 receptor type 1 (IL-1R1) signaling in H1N1 influenza virus infection. Methods IL-1R1 knockout ( IL-1R1-/-) mice and wild type ( WT) mice were infected intranasally with 2×104 TCID50(50% tissue culture infective dose) of influenza virus H1N1 PR8. Changes in clinical signs, survivals and bodyweights of those mice were monitored daily for 14 consecutive days. Three mice from each group were sacrificed at 3, 7 and 14 days post infection (d. p. i), from which whole lungs were harvested. A part of the lobes was fixed in 4% paraformaldehyde for histopatho-logical assessment and the rest were split and stored at-80 centigrade for further analysis. Real-time quanti-tative PCR and cytometric bead array ( CBA) were performed to detect viral loads in lungs and inflammatory cytokines in supernatants of lung homogenates. Results The mice in both groups showed severe symptoms after the infection of PR8. The maximum bodyweight loss of IL-1R1-/- mice [(24. 22±0. 80) % at 8 d. p. i] was lower than that of WT mice [(28. 03±1. 51)% at 9 d. p. i] (P<0. 05). The IL-1R1-/- mice with PR8 infection showed a higher survival rate (90%) as compared with that of the control group (40%) (P<0. 05). No statistical differences in virus loads were observed between the two groups at 3, 7 and 14 d. p. i. The lung weight to body weight ratio of IL-1R1-/-mice [(1. 42±0. 03) %] was lower than that of WT mice [(1. 79±0. 08) %] at 3 d. p. i (P<0. 05). Pathological changes in IL-1R1-/- mice were less severe than those in WT mice. CBA detection assay revealed that the proinflammatory cytokines in lungs of IL-1R1-/-mice were less than those in WT mice. Conclusion IL-1R1 signaling plays a pathogenic role in mice infec-ted with 2×104 TCID50 of influenza virus PR8 by promoting inflammatory responses.

OBJECTIVETo explore the effect of combination of Epimedii Folium and Ligustri Lucidi Fructus on osteoporosis rats induced by retinoic acid.

METHODSixty three-month-old male Wistar rats were randomly divided into the normal control group, the model group, the Epimedii Folium group, the Ligustri Lucidi Fructus group, the combination group of Epimedii Folium and Ligustri Lucidi Fructus and the raloxifene group. The osteoporosis model was established through oral administration with retinoic acid for two weeks. Meanwhile, all of treatment groups were administered with corresponding drugs for three weeks. The contents of serum calcium (Ca), phosphorus (P), alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (StrACP) were detected, and the pathomorphological changes of femurs were observed.

RESULTThe model control group showed much lower contents of serum Ca and P than the normal control group, but with significantly higher AKP and StrACP activity than the normal control group. The femoral head area showed reduced, narrow and sparse trabecular bones, with typical osteoporosis-like changes. Compared with the model control group, all of treated groups showed significant increase in Ca and P contents in serum, and down-regulate AKP and StrACP levels, while trabecular bones became more and wider, and densely interweaved as a reticular formation. Among them, the combination group showed the most significant effect.

CONCLUSIONEpimedii Folium and Ligustri Lucidi Fructus could effectively correct the abnormal bone metabolism and improve pathological conditions of bone tissues, so as to show the anti-osteoporosis effect. The combined application of the two drugs showed a better efficacy.

Acid Phosphatase ; blood ; Alkaline Phosphatase ; blood ; Animals ; Body Weight ; drug effects ; Calcium ; blood ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Epimedium ; chemistry ; Femur ; drug effects ; pathology ; Isoenzymes ; blood ; Ligustrum ; chemistry ; Male ; Osteoporosis ; blood ; chemically induced ; drug therapy ; pathology ; Phosphorus ; blood ; Rats ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase ; Tretinoin ; adverse effects

ObjectiveTo explore the relationship between chronic kidney disease (CKD) and cerebral small vessel disease (SVD) in elderly patients. MethodsOne hundred and fifty-two elderly male CKD patients for experimental group and 158 elderly male for control group were recruited. Demographic data and vascular risk factors were recorded. White matter lesion (WML) was semi-quantitatively assessed by cerebral magnetic resonance imaging (MRI), and lacunar infarction (LI) was also calculated. Results(1) The prevalenees of hypertentsion and diabetes mellitus were higher in elderly CKD patients than those in control group (30. 9% vs. 19.0%, 23.7%vs. 14.6%;both P～0. 05). (2) The percentages of grade 2 and grade 3 WMLs were higher in elderly CKD patients than those in control group (34.9% vs. 24.1%, 25.7% vs. 16.5%;both P<0.05). Prevalence of LI was higher in elderly CKD patients than that in control group (45.4% vs.25.3% ,X2= 13. 70, P<0. 05). The similar Resultswere also obtained except for control subjects with hypertension and diabetes mellitus. (3) The logistic regression analysis showed that age, hypertension and low glomerular filtration rate (GFR) were closely associated with SVD in elderly CKD patients. ConclusionsHypertention and diabetes mellitus are risk factors for CKD in elderly patients. SVD is associated with CKD, and age, hypertension and low GFR may be risk factors for SVD in elderly CKD patients.

Objective To analyze the complications after procedure for prolapse and hemorrhoids (PPH) in treating severe mucocutaneous hemorrhoids.Methods The clinical data of 550 patients with severe mucocutaneous hemorrhoids who were admitted to the Hangzhou Hospital of Nanjing Medical University from January 2005 to December 2013 were retrospectively analyzed.All the patients were treated by PPH or PPH + external hemorrhoids resection.The postoperative complications were recorded and assessed.Patients were followed up via outpatient examination and telephone interview till June 2014.Results Operation was successfully carried out on all the 550 patients.The operation time and duration of hospital stay were (15 ± 10)minutes and (4.2 ± 1.8)days,respectively.A total of 445 patients were followed up for 6-60 months,with the median time of 28 months.The overall incidence of complications was 14.61％ (65/445).Forty-five patients were complicated with anal pain,12 with hemorrhoid recurrence,7 with bleeding,5 with dysporia,4 with anastomotic stenosis and 2 with rectal and perianal infection.Of the 12 patients with hemorrhoids recurrence,5 was alleviated by medication and 7 received PPH for the second time.Of the 4 patients with anastomotic stricture,the condition of 2 patients were alleviated by anus dilatation,and the condition of 1 patient was alleviated by balloon dilatation under enteroscope,1 patient received circular incision of the stricture because of severe stricture and failed of conservative treatment.Of the 2 patients with rectal and perianal infection,1 was alleviated by intravenous and local medication,and the other patient who was complicated with anal fistula was alleviated by surgery.The condition of other patients was improved after symptomatic treatment.Conclusions PPH is effective and safe for the treatment of severe mucocutaneous hemorrhoids.The main complication is anal pain,and it could be cured or alleviated by symptomatic treatment.

Adult ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Imaging, Three-Dimensional ; Male ; Maxillofacial Injuries ; diagnostic imaging ; surgery ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; methods ; Skull Fractures ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

Objective To explore the correlation between NF-κB signaling pathways activated by IL-18 in CD4+ T cells and the pathogenesis of PBC.Methods We detected the expression of IL-18 mRNA in PBMCs,IL-18 level in plasma,receptor IL-18R on surface of CD4+ T cell,proliferation rate of CD4+T cell and its NF-κB signaling pathway protein IκBα and NF-κB p65 by qRT-PCR,ELISA,flow cytometry,MACS and Western blot on 32 cases of patients with PBC (PBC group) and 32 healthy people (control group) in Guizhou provincial people′s hospital.Results The level of IL-18 in PBC group was significantly higher than that in control group (P<0.05).The relative expression of IL-18 mRNA in PBC group was significantly higher than that in control group (P<0.05).The percentage of CD4+T cells expressing IL-18Rα in PBC group was higher than that in control group (P<0.05).The proliferation rate of CD4+T cells stimulated by IL-18 in PBC group was significantly higher than that in healthy control group (P<0.01).The relative expression levels of NF-κB p65 protein were up-regulated in IL-18,and the expression of IκBα protein in each group was significantly increased,especially in PBC group (P<0.01).Conclusion IL-18 can activate NF-κB signal pathway in CD4+ T cells and participate in the pathogenesis of primary biliary cirrhosis.