Objective To study the effects of Poly(I:C) on lung adenocarcinoma A549 cells viability and illuminate the mechanism of Poly (I:C)-induced apoptosis in A549 cells.Methods A549 cells were transfected with the complex of Poly(I:C) and lipofectamine 3000.The viability of A549 cells was tested by methyl thiazolyl tetrazolium (MTF) method.The apoptotic cells were tested by flow cytometry.The caspase proteins were tested by Western blotting and the expressions of interferon-β (IFN-β) and CXCL-10 were assayed by real-time PCR.After employing the pan-caspase inhibitor Z-VAD-FMK and caspase-8 inhibitor Z-IETD-FMK,the variation of Poly (I:C) proapoptosis in A549 cells was observed.RNA interfering experiments were employed to knock down melanoma differentiation related antigen 5 (MDA5) or retinoic acidinduced gene Ⅰ (RIG-Ⅰ),and the above indexes were tested.Results The viability of A549 cells was significantly reduced to 74.92％ ±--6.24％ after 200 ng/ml Poly (I:C) transfection compared with that before transfection (95.32％ ± 3.05％,t =2.883,P =0.041).The apoptotic rates induced by 100,200,400 ng/ml Poly(I:C) were 9.97％-± 0.88％,23.63％-± 1.41％,32.57％-± 2.39％,respectively.All of them were higher than that in the control group (0.74％-± 0.15％),with significant differences (t =4.489,P =0.002;t =11.616,P =0.000;t =16.932,P =0.000).Besides,the death receptor pathway proteins such as TNF-related apoptosis inducing ligand (TRAIL),cleaved-caspase-8 and cleaved-caspase-3 increased obviously.MDA5/RIG-Ⅰ pathway was also activated dramatically and the expressions of IFN-β,CXCL-10 were significantly up-regulated.The apoptotic rates reduced to 3.17％ ± 0.66％,5.35％ ± 0.64％ with pancaspase inhibitor Z-VAD-FMK and caspase-8 inhibitor Z-IETD-FMK pretreatment,compared with the control group (15.87％ ±0.93％),and the differences were statistically significant (t =8.643,P =0.001;t =6.824,P =0.002).Moreover,the expressions of TRAIL,IFN-β and CXCL-10 induced by Poly (I:C) were inhibited with MDA5 or RIG-Ⅰ depletion.Conclusion Poly(I:C) can reduce the survival rate of A549 cells and promote the apoptosis mainly by activating the death-receptor pathway mediated by MDA5/RIG-Ⅰ probably,which may involve in IFN-β,CXCL-10.

Objective To investigate the selective oncolytic role and antitumor action of a novel recombinant adenovirus containing E1A and IL-24 on hepatocellular carcinoma cell(HCC). Methods The recombinant adenovirus expressing IL-24 (Ad. HS4. AFP. E1A/IL-24) was constructed by using modified human alpha-fetoprotein (HS4-AFP) promoter to drive adenovirus E1A gene and II-24 gene.Cell Counting Kit-8 were performed to test the selective cytotoxicity of the virus in hepatocellular carcinoma cell lines SMMC-7721, Hep3B, MHCC97-H and hepatocyte cell line L02 . The mRNA and protein expression of IL-24 gene were detected by RT-PCR and western blot. Cell growth curves and Annexin V/PI assay were used to study cell proliferation and apoptosis of MHCC97-H. The anti-metastatic effects of the recombinant adenovirus were evaluated in cell adhesion, migration, and cell motion. Matrix metalloproteinase-2 (MMP-2) expression was examined by RT-PCR and zymography.Results Selective replications of Ad. HS4. AFP. E1A/IL-24 adenovirus were observed in over expression AFP cell line MHCC97-H, a highly metastatic potential HCC cell line but not in hepatocyte cell line L02. The mRNA and protein of IL-24 were also over expressed in MHCC97-H. This recombinant adenovirus also showed the significant oncolytic action on MHCC97-H but not on L02 (P＜0. 05). Besides, the recombinant adenovirus significantly inhibited MHCC97-H metastatic potential such as cell adhesion, migration and invasion as well(P＜0.01). Conclusion The selective oncolytic adenovirus expressing E1A and II-24 has a selective antitumor effect and play an inhibitory role in metastasis of HCC.

To make further study on the mechanism of needling based on the concept of needle-acupointomics.This article aims to reveal the biologic basis and interpret the scientific methods of needling,and to optimize needling prescriptions.The article make a standardized and normalized study on key techniques in clinical therapeutic methods,for the purpose of putting scientific mechanism into the practical usage,and better serving for clinical practice.

Objective To obtain scientific data for control of iodine deficiency disorders(IDD) by reviewing the surveillance information of school children intelligence quotient(IQ) after the implementation of universal salt iodization. Methods One thousand five hundred and eighty children were selected from 11 primary school in Dalian city of Liaoning province during 2006 to 2009. IQ was measured by Combined Raven Test-C2(CRT-C2) in China. Groups of IQ were classified as outstanding(≥ 130), excellent (120- 129), above average (110- 119), average (90 - 109), below average(80 - 89), margin(70 - 79), low(≤69). Urinary samples of children were collected randomly. Urinary iodine were determined by As-Ce catalytic spectrophotometry. The growth characteristics of IQ were analyzed according to surveillance year and born year. Results The average IQ of children aged 8-10 were 110.4 ± 14.0,112.5 ± 12.4,117.2 ± 11.4,116.2 ± 12.6, respectively, increased year by year from 2006 to 2009. Its average annual increase from 2007 to 2009 were 2.1,3.4,1,9 compared with the IQ in 2006 respectively. The medians of urinary iodine were 224.7,266.7,222.1 μg/L from the year 2007 to 2009, respectively, which were all between 200 - 300 μg/L and can be classified as more than adequate level. The average IQ of children born during the year of 1994 to 2000 were 106.7 ± 13.0,108.1 ± 13.9,108.5 ± 13.4,111.3 ± 14.3,113.6 ± 12.5,115.3 ± 12.3,119.8 ± 11.2, respectively. Its average annual increase from 1995 - 2000 were 1.4,0.9,1.5,1.7,1.7,2.2 compared with the IQ born in 1994 respectively. The ratio of IQ in margin group and low group were all below 2% ; the ratio was increasing in excellent group and outstanding group and decreasing in average group and below average group significantly year after year(x2 = 52.471,34.329,66.483,11.148, all P<0.01). Conclusions Universal salt iodization improves IQ scores. IQ index should be brought into the surveillance project and put in use in IDD control.

Vincristine (VCR) is mainly used to treat acute lymphocytic leukemia, Hodgkin and non-Hodgkin lymphoma in clinic with definite therapeutic effect. But the obvious neurotoxicity and local stimulation of which limit its clinic use. In order to increase the lymph targeting to enhance the curative effect and to lower the adverse reaction of VCR, the VCR loaded transfersomes (VCR-T) were prepared with dry-film and ultrasonic dispersing methods, and the corresponding pharmaceutical properties, pharmacokinetical characteristics and the targeting ability were studied. The average particle size of VCR-T prepared was 63 nm with an entrapment ratio of 59%. The in vitro transdermal research with modified Franz cell showed that VCR-T permeated through the skin in accordance with polynomial equation, and with an accumulation permeation percentage of 67.4% up to 12 h. An HPLC method was utilized to determine the pharmacokinetics and tissue distribution of VCR. Compared with the iv injection of VCR solution, the retention time of VCR in blood was extended by 12 times with VCR-T, and the targeting index in rat lymph was increased by 2.75 times. As a result, transfersomes could penetrate the skin and enter into the systemic circulation carrying VCR with good lymph targeting ability, which makes it probably a new lymphtic targeting drug delivery system.
Administration, Cutaneous ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; blood ; pharmacokinetics ; Area Under Curve ; Drug Delivery Systems ; Liposomes ; chemistry ; Lymph Nodes ; metabolism ; Male ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Skin Absorption ; Spleen ; metabolism ; Surface-Active Agents ; chemistry ; Tissue Distribution ; Vincristine ; administration & dosage ; blood ; pharmacokinetics

OBJECTIVETo study the clinical features, treatment and diagnosis of parapelvic cyst.

METHODSTwenty-three patients of parapelvic cyst of the kidney were reviewed retrospectively. Fourteen cases (61%) complained of lumbar pain or discomfort, and 4 patients (17%) accompany hematuria and hypertension.

RESULTSIn 15 patients receiving surgery, 2 were treated by nephrectomy, one by radical nephrectomy for misdiagnosis. Postoperative diagnosis confirmed a cyst. Eight patients were treated conservatively for cyst being small and without clinical symptoms. Nineteen cases were followed up for 0.5 - 12.0 years.

CONCLUSIONSUltrasonography and CT scan are the main diagnostic methods. Enhanced CT is extremely helpful in differential diagnosis of hydronephrosis. Surgical management is suitable for big cysts, lumbar pain, hematuria, hypertension and other complications.

Adult ; Aged ; Diagnosis, Differential ; Female ; Humans ; Kidney Diseases, Cystic ; diagnosis ; etiology ; surgery ; Kidney Pelvis ; Male ; Middle Aged ; Prognosis ; Tomography, X-Ray Computed

OBJECTIVEThe nose is composed of several delicate subunits, some of which are difficult to reconstruct if they have been injured. The paper presents the microsurgical technique to repair the nasal subunit defects with the free combined preauricular and auricular flap which well match the nasal tissues in texture, contour and color.

METHODSThe nasal subunit defects were repaired with the combined preauricular and auricular flap which were vascularized by the superficial temporal vascular system. The flap was harvested from the contralateral preauricular and the region of helix crus. The superficial temporal vessels were anastomosed to facial vessels via the vascular grafts harvested from the lateral circumfluent femoral vessels, which were about 10 to 14 cm in length. The helix crus of donor ear was reconstructed with the post-auricular flap.

RESULTS28 cases were treated, including 3 cases of nasal tip defects, 9 cases of combined nasal alar and sidewall defects, and 16 cases of nasal alar defects. In these cases, the size of the subunit defects varied from 2.5 cm x 1.5 cm to 4 cm x 2.5 cm. 27 cases were successfully repaired with satisfactory results. The contour of reconstructed helix crus in donor site was acceptable. No walking dysfunction of the donor thigh was complained. There is one failed case, and the possible reason is the insufficient blood perfusion to the flaps, which may due to the patient's longtime-smoking status and the hypertension.

CONCLUSIONSThe technique of free combined preauricular and auricular flap is ideal for the reconstruction of nasal subunit defects.

Adolescent ; Adult ; Ear Auricle ; surgery ; Female ; Humans ; Male ; Microsurgery ; Middle Aged ; Nose ; abnormalities ; Nose Deformities, Acquired ; surgery ; Rhinoplasty ; methods ; Skin Transplantation ; Surgical Flaps ; Young Adult

Acetylcysteine ; therapeutic use ; Animals ; Liver Cirrhosis, Experimental ; drug therapy ; Magnesium Compounds ; therapeutic use ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the diagnosis, treatment, and prognosis of prostatic malignant mesenchymal tumors (PMMT).

METHODSWe retrospectively analyzed the clinical and follow-up data about 20 cases of PMMT and reviewed the literature relevant to the diagnosis, treatment, and prognosis of the disease.

RESULTSBased on the results of pathology and immunohistochemistry, the 20 PMMT cases included leiomyosarcoma (n = 7), rhabdomyosarcoma (n = 5), prostatic stromal sarcoma (n = 3), chondrosarcoma (n = 1), and undifferentiated PMMT (n = 4). Twelve of the patients were treated by radical prostatectomy (3 concurrently by sigmoid colostomy and 1 by cystostomy), 2 by pelvic tumor resection following arterial embolization, 1 by total pelvic exenteration, 1 by colostomy with pelvic lymph node biopsy, and 4 by conservative therapy because of metastasis to the lung, pelvis and bone. Of the 20 patients, 9 died of systemic metastasis within 3 months after treatment, 3 died at 6, 7, and 14 months, respectively, 3 survived with tumor for 5, 11, and 12 months, respectively, 2 survived without tumor for 12 and 24 months so far, all subjected to periodic chemotherapy postoperatively, and 3 lost to follow-up.

CONCLUSIONPMMT is a tumor of high malignancy and rapid progression, for which transrectal ultrasound-guided biopsy remains the main diagnostic method. The clinical stage of the tumor is an important factor influencing its prognosis and the survival rate of the patients can be improved by early diagnosis and combined therapy dominated by radical prostatectomy.

Combined Modality Therapy ; methods ; Humans ; Immunohistochemistry ; Male ; Mesenchymoma ; mortality ; pathology ; therapy ; Prognosis ; Prostatectomy ; Prostatic Neoplasms ; mortality ; pathology ; therapy ; Retrospective Studies

OBJECTIVETo investigate effect of AP-1 and Ets binding site adjacent to matrix metalloproteinase-9 (MMP-9) promoter on activation of MMP-9 transcription of nasopharyngeal carcinoma cells transfected with EBV-encoded latent membrane protein 1 (LMP1), and to ascertain if cross-talk between c-Jun and Ets1 is involved in LMP1-regulating expression of MMP-9.

METHODSSite-directed mutagenesis technique was used to establish a series of mutants, including MMP-9-CAT-Ets(-540)mt, MMP-9-CAT-AP-1(-533)mt and MMP-9-CAT-AP-1(-533)/Ets(-540)mt. After the mutants were transfected into LMP1-expressing NPC HNE2 cells regulated by Tet-on system (pTet-on-LMP1 HNE2), CAT activity of these mutants were assayed with induction of LMP1. With blockade of c-Jun or Ets1 antisense oligonucleotides, the activity of MMP-9 induced by LMP1 was assayed with gelatin zymography.

RESULTSThe CAT activity of MMP-9-Ets(-540)mt-CAT, MMP-9-AP-1(-533)mt-CAT, MMP-9-AP-1(-533)/Ets(-540) mt-CAT decreased significantly compared to MMP-9-CAT wt. After blockade with c-Jun or Ets1 antisense oligonucleotides, activity of MMP-9 induced by LMP1 decreased significantly, especially with combined blockade of c-Jun and Ets1.

CONCLUSIONThe results suggest that transcription factor AP-1 and Ets play an crucial role in activation of MMP-9 transcription induced by LMP1, and cross-talk between c-Jun/Ets1 is involved in expression of MMP-9 mediated by LMP1.

Herpesvirus 4, Human ; genetics ; Humans ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Nasopharyngeal Neoplasms ; metabolism ; virology ; Proto-Oncogene Protein c-ets-1 ; genetics ; Proto-Oncogene Proteins c-jun ; genetics ; Transfection ; Tumor Cells, Cultured ; Viral Matrix Proteins ; genetics