Apoptosis of PK-15 cells induced by Foot-and-Mouth Disease Virus (FMDV) in vitro was reported in this paper. Typical cell apoptosis was detected by use of Hoechst 33258 fluorescence probe, agarose gel electrophoresis and in situ end-labeling (TUNEL). After PK-15 cells were infected by titration of 4.8 lg TCID50/mL FMDV for 32 h, apoptosis characteristics of nuclear condensation, fragmentation, accompanied by apoptotic bodies formation (Hoechst 33258 staining), 180-200 integer-fold sized pieces DNA Ladders (agarose gel electrophoresis) and strong green fluorescence dots (TUNEL) were all exhibited, and cell apoptosis was approximately 20%. In addition, the quantitative analysis of apoptosis in PK-15 cells induced by FMDV showed that apoptosis was correlated with infection of virus, and it was also time-dependent. Results indicate that FMDV can induce apoptosis of host cells and apoptosis plays an important role in the cytopathogencity effect of FMDV.

Child, Preschool ; Female ; Humans ; Trichothiodystrophy Syndromes ; etiology

Objective　To investigate the molecular regulation of G1 arrest of mouse thymocytes induced by ionizing radiation. Methods　Cell cycle was analyzed by flow cytometry (FCM) following staining of cells with proidium iodide. Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression. Results　It was demonstrated that G1 phase of mouse thymocytes increased significantly at 12h after whole body irradiation (WBI) with the doses of 0.5, 1.0 and 2.0 Gy, and at 24h following 2.0Gy exposure, measured by FCM. In the time course experiment, it was found that G1 phase of thymocytes increased significantly at 4h, reached a peak level at 24h and came down toward 48h after WBI with 2.0Gy X-rays. The results also showed that after 2.0Gy exposure, the expression of proteins in mouse thymocytes increased significantlty from 1h to 8h for p53, for p21 from 4h to 48h, and for MDM2 at 4h and 8h, measured by FCM. But no change was found for GADD45 protein expression. Conclusion　These results suggest that G1 arrest could be induced by a single dose of 0.5 Gy, 1.0Gy or 2.0Gy, and its molecular control might be established through the p53-p21 pathway.

Background Research demonstrated that mitochondrial pathway plays a key role in cell apoptosis.Purendan supermicropowder(PRD),a traditional Chinese medicine,may be a potentially effective therapy for neuron apoptosis in diabetic retina.Objective This study was carried out to investigate the effects of PRD on aldose reductase(AR)activity,neuron apoptosis and mitochondrial pathway in retina of diabetic rat.Methods Thirty-six clean male Wistar rats were randomly divided into normal control group,diabetes model group,PRD treatment group randomly and 12 rats for each group.The diabetes models were established by intraperitoneal injection of 25 mg/(kg · d)streptozotocin(STZ)for 3 consecutive days,and blood glucose ≥ 16.7 mmol/L was taken as the standard.PRD solution of 1.8 g/(kg · d)was lavaged in 12 models for 3 months.The eyeballs were enucleated for the preparation of retinal tissue homogenate and slice.AR activity in the retina was detected by ultraviolet spectrophotometry,and neuron apoptosis in retina was assayed by TUNEL staining.Western blot was used to assess the expressions of bcl-2,bax,cyt-c and caspase-3 protein in the retina.The use of animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Committee(Version 1988).Results Statistically significant differences were found in AR activity and AI among the normal control group,diabetic group and PRD groups(F=90.115,165.540,P＜0.01),and those of diabetic group were evidently higher than the normal control group and PRD group(P＜0.01,P＜0.01).The positive TUNEL cells mainly located in inner nuclear layer and retinal ganglion cell layer.The expressions of bax,cyt-c,caspase-3,bcl-2 and bcl-2/bax in retina were obviously different among these three groups(F =51.332,41.262,25.888,38.564,47.870,P＜0.01),and the expression of bax,cyt-c and caspase-3 protein in diabetic group evidently elevated in comparison with the normal control group and PRD group(t ＝ 10.32,11.04,6.91,P ＜ 0.01)and the expressions of bcl-2 protein and bcl-2/bax value were significantly lower in diabetic rats than in the normal control rats(t =18.05,12.23,P＜0.01).AR activity by AI of retina,the expressions of bax,cyt-c and caspase-3 proteins in retina were obviously lower in PRD group than in diabetes model rats(P ＜ 0.01),and the expression of bcl-2 protein and bcl-2/bax value were significantly higher in PRD group than in diabetes group(P＜0.01).Conclusions PRD can protect retina against the damage caused by high glucose by suppressing AR activity by downregulating the expressions of bax,cyt-c,caspase-3 proteins,increasing the expressions of bcl-2 protein in retina of diabetic rats and further inhibiting the mitochondrial pathway and reducing cell apoptosis in retina of diabetic rats.

ObjectiveTo investigate the application of intraoperative Computed Tomograph (CT) using in surgery for complex acetabular fractures.MethodsFrom June 2008 to December 2010,14 patients (9 males,5 females; with the mean age of 45.1 years; range,28-62 years) with complex acetabular fractures were operated using intraoperative CT.Preoperative radiotherapy and CT scan were adopted to evaluate the fractures.Three dimensional reconstruction based on CT scan was used to mimic surgery.The surgery approach and the type of internal fixators were noted.Intraoperative C-arm and CT scan were used to evaluate the fractures reduction respectively.Decision of additional reduction was made by surgeons according to above mentioned methods respectively and the results were noted.Comparing to preoperative design,the change of surgery plan were noted.Overall time,frequency and radiation dose of intraoperative CT scan were also noted.ResultsAll patients in this study received average 2.7 times of intraoperative CT scan.Mean time of CT scan was 40.4 min and the overall dose of radiation was 47.2 mGy.Decision of additional reduction was made in 3 cases according to C-arm radiography and 4 cases according to CT scan (above mentioned 3 cases were included).The change of surgery plan was made in one case.In postoperative radiography evaluation according to Matta's score system,anatomical reduction were achieved in 8 cases,imperfect reduction in 3 cases and poor reduction in 3 cases.ConclusionIntraoperative CT scan increases the radiation time and dose of patients dramatically.When used to evaluate fracture reduction intraoperatively,it can't take the advantage of traditional C-arm radiography.When delicate preoperative plan is made with radiography and three dimensional reconstruction based on CT data,the efficiency of intraoperative CT scan for complex acetabular fractures are to be discussed.

BACKGROUNDOne stage transanal Soave pull-through procedure (TSPP) is a recent popular operation in the treatment of Hirschsprung's disease (HD). With no visible scar and a short hospital stay, it is well accepted by surgeons and mothers. In the conventional Soave procedure, a long rectal muscular cuff left for anocolic anastomosis might increase the incidence of postoperative enterocolitis and constipation. This study presents a modified transanal Soave pull-through procedure (MTSPP) which includes an oblique mucosectomy and an oblique anastomosis with a short split muscular cuff.

METHODSA review of two groups of HD patients was made: 112 underwent conventional transanal Soave procedure from 1999 to 2001 (group 1) and 140 underwent modified transanal Soave procedure from 2002 to 2004 (group 2). A comparison was made between the two groups on operative data and postoperative complications. The data included: age at the operation, operating time, blood loss, time to feeds and hospital stay, occurrence of postoperative enterocolitis or constipation, need for anal dilatation, postoperative bowel function and perianal skin problems.

RESULTSThere was no significant difference between two groups with respect to age, gender, length of colon resected, operating time, blood loss and hospital stay. However occurrence of postoperative enterocolitis, constipation, anastomotic stricture and time needed for anal dilatation were evidently less in group 2 (MTSPP). The mean operating time in group 1 was (106 +/- 39) minutes with a range of 60 to 170 minutes; in group 2 was (101 +/- 36) minutes with a range of 66 to 190 minutes. The average length of the bowel resected in group 1 was (24 +/- 7) cm, range 15 to 58 cm; in group 2 was (26 +/- 8) cm, range 15 to 70 cm. Two patients, one in each group, required laparoscopic assistance because of long aganglionic colon. Another patient in group 2 required laparotomy because of total colonic aganglionosis. Postoperative complications in group 1 included: temporary perianal excoriation in 34 patients (26 were < 3 months of age), enterocolitis in 21, anastomotic stricture in 11, recurrent constipation in 12, cuff abscess in 1, anastomosis leak in 1, soiling in 3 and rectal prolapse in 1. In group 2 post operative complications included: transient perianal excoriation in 37 patients (30 were < 3 months of age), enterocolitis in 13, anastomotic stricture in 5, recurrent constipation in 6, anastomotic leak in 1, adhesive bowel obstruction in 1 and soiling in 4. Complete bowel continence was found in 97 children (86.6%) in group 1 and in 129 children (92.1%) in group 2 at one year followup after operation.

CONCLUSIONSModified transanal Soave pull-through procedure for HD with oblique mucosectomy and anastomosis and a short split muscular cuff is a safe and feasible operation with low incidence of postoperative complication. It is an encouraging improvement of the conventional transanal Soave pull-through procedure. MTSPP is a preferable choice in the surgery of HD.

Adolescent ; Child ; Child, Preschool ; Digestive System Surgical Procedures ; methods ; Enterocolitis ; etiology ; Female ; Hirschsprung Disease ; surgery ; Humans ; Infant ; Infant, Newborn ; Male ; Minimally Invasive Surgical Procedures ; Postoperative Complications ; etiology

Hepatitis, Autoimmune ; immunology ; therapy ; Humans ; Immunotherapy, Adoptive

OBJECTIVETo study the correlation between the expression of Egr-1 and NF-kappaB and the up-regulation of TNF-alpha and TGF-beta1 in macrophages after stimulation by silica in-vitro.

METHODSMacrophages were treated with antibodies against Egr-1 and NF-kappaB and antisense oligonucleotides. The level of TNF-alpha protein in the cell supernatant was then measured using enzyme-linked immunoadsorbent assay (ELISA). The expression of TGF-beta1 protein was detected by immunocytochemistry. The expression of TNF-alpha and TGF-beta1 mRNAs was also monitored by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSCompared with silica-stimulated macrophages untreated with antibodies, the cells treated with 10 micro g/ml of Egr-1 or NF-kappaB antibodies were associated with reduced expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05). Compared with silica-stimulated untransfected group, the antisense group was associated with obvious reduction in the expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05).

CONCLUSIONThe expression of TNF-alpha and TGF-beta1 mRNAs and proteins are associated with activation of Egr-1 and NF-kappaB in macrophages, after stimulation by silica. It is possible that the corresponding antibodies and antisense oligonucleotides may become a potential therapeutic tool in the management of silicosis in the future.

Animals ; Antibodies ; immunology ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; immunology ; Early Growth Response Protein 1 ; Immediate-Early Proteins ; genetics ; immunology ; Macrophages ; cytology ; metabolism ; Mice ; NF-kappa B ; genetics ; immunology ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Silicon Dioxide ; pharmacology ; Silicosis ; etiology ; Transcription Factors ; genetics ; immunology ; Transforming Growth Factor beta ; biosynthesis ; genetics ; Transforming Growth Factor beta1 ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

BACKGROUNDThe treatment of patients with small cell lung cancer (SCLC) is based on chemotherapy. However, the treatment is limited by the development of drug resistance. Emodin has been shown to exhibit an anti-cancer effect. But the molecular mechanism remains unclear. This study was conducted to investigate the effect of emodin on the gene expression profile changes in SCLC NCI-H446 cells.

METHODSNCI-H446 cells were treated with emodin and cell viability was determined by MTT assay. Cell apoptosis was determined by both flow cytometry and caspase-3 activity assay. The effect of emodin on the gene expression profile of NCI-H446 cells was analyzed using cDNA microarray. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to validate the microarray results.

RESULTSEmodin suppressed viability, induced apoptosis and changed cell cycle of NCI-H446 cells. Among the 1262 genes, 10 genes were up-regulated and 8 genes were down-regulated more than 2 folds in NCI-H446 cells when compared with the control cells after treatment with emodin for 12 hours, while 12 genes were up-regulated and 24 genes were down-regulated after treatment with emodin for 24 hours. These genes were involved in metabolism, signal transduction, transcription regulation, cytoskeleton organization, immune response, transport, protein synthesis, cell cycle control, cell adhesion and RNA processing. The RT-PCR results were consistent with those obtained by the microarray.

CONCLUSIONSEmodin affects the expression of genes involved in various cellular functions and plays important roles in cell apoptosis, tumor metastasis and chemotherapy-resistance, which suggests emodin might become an effective chemopreventive or chemotherapeutic agent for SCLC.

Apoptosis ; drug effects ; Carcinoma, Small Cell ; drug therapy ; metabolism ; pathology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Emodin ; pharmacology ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Lung Neoplasms ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction

Objective To study the efficacy and safety of T-614 in treating rheumatoid arthritis(RA). Methods Two hundred and eighty patients with active RA were randomly allocated to 3 groups:T-614 50 mg each day,25 mg each day or placebo.Clinical and laboratory parameters were analyzed at baseline,2,4,6,12, 18 and 24 weeks.Results The ACR response rate was significantly higher in the T-614 treatment group com- pared with the placebo group during the first 6 weeks.After 24 weeks,25 mg/d,50 mg/d dosage group and the placebo group showed 39.1%,61.3% and 24.2% in ACR20,23.9%,31.2% and 7.4% in ACR50 respectively.A time-response in ACR response after 24 weeks was observed,with clear superiority of the 25 mg/d and 50 mg/d dosage groups compared to the placebo,and 50 mg/d dosage group compared to 25 mg/d dosage group(P