[Objective]To evaluate the expression of enhancer of zeste homolog 2(EZH2),Ki-67 and intratumoral microvaseu?lar density(iMVD)in human colorectal cancer(CRC),and discuss their relationship with the biological behavior and prognosis.[Methods]The expression of EZH2,Ki-67 and iMVD(labeled by CD34 protein)was measured with immunohistochemical MaxVision method in CRC patients followed up over 5 years.[Results]Positive expression rates of EZH2,Ki-67 and iMVD were 40.91%(45/110),57.27%(63/110)and 46.36%(51/110),respectively. EZH2 expression was positively correlated with distant metastasis(P=0.024). Ki-67 expression was positively correlated with tumor size,infiltration depth,AJCC stage and differentiation(P=0.033~0.015). The iMVD expression was positively correlated with infiltration depth ,AJCC stage and differentiation (P=0.016~0.034). Spearman correlation analysis showed that EZH2 expression was positively correlated with Ki-67 expression(r=0.195,P=0.041), whereas negatively correlated with iMVD expression. Ki-67 expression had a positive correlation with iMVD(r=0.213,P=0.025). Multivariate analysis suggested that the EZH2 expression was an adverse independent factor for survivals of CRC patients(HR 1.965, 95%CI:1.019-3.789,P=0.044 for OS).[Conclusion]EZH2 expression was significantly correlate with the proliferation of tumor cells and may plays an important role in the metastasis and prognosis of CRC.

Objective To investigate the water quality and running status of the dringking water improving project in Xuchang,Pingdingshan and Nanyang,thus to provide basis for scientifically evaluating social effects.Methods Stratified sampling was used in the levels of counties and townships and villages,with the number of checked counties greater than or eaquel to half of the projected counties(13/24),the checked townships more than 40 percent of the projected townships(42/102),the checked villages greater than or eaquel to 30% of the projected villages(56/186).We listened their statement,reviewed the documents,examined carefully the water projects before we made the appraisal.Results More than 95%(129/136)of the projects had water quality reports,among which 90%(122/136)were provided by county level centers for disease prevention and control,75% (102/136) of water samples were collected by centers for disease prevention and control,80%(109/136)ofcounties organized an acceptance check-up group,92%(125/136)projects well preserved water source.Conclusions The dringking water improvement projeets in countryside are basically normal in terms of construction and management,the water supplied is qualified,so the expected goal is achieved.However,duties of each department are not explicit and the communication is inadequate,so collaboration should be reinforced.

Objective To observe toxic effect of deoxynivalenol(DON)on articular cartilage and synovium of New Zealand rabbits's knee ioints.Methods Fifteen male rabbits were divided randomly into 3 groups:control, high-dosage,and low-dosage group.In high-dosage and low-dosage group,saline solution of DON was injected with a dose of 0.10 and 0.05 ms/kg every 48 h into ear vein of rabbits.Specimen of articular cartilage and synovium were through pathologY methods,and IL-1β,TNF-α,NO levels were assayed in joint liquid,after 20 days. Results Morphological changes were observed, such as synovium inflammative infiltration, chondrocytes deformation and necrosis under light microscope.The levels of IL-1β,TNF-α and NO had statistical significance in comDarison between 3 grouPs(F=19.396,18.195,22.136,P＜0.05).The levels of IL-1β,TNF-α and NO were significantly higher(all P＜0.05),high-dosage[(0.451±0.091),(0.575±0.122)μg/L;(70.27±11.53)μmol/L] and low-dosage group[(0.295±0.107),(0.387±0.131)μg/L;(45.32±12.24)μmol/L]compared with control ((0.1 13±0.049),(0.138±0.087)μg/L;(23.56±9.35)μmoL/L],and high-dosage compared with low-dosage group Conclusions DON results in articular and synovial impairment,which has the symptom similar to osteoarthritis. DON probably causes osteoarthritis.

OBJECTIVETo investigate the inhibitory effects of CaMKIIN on acute myeloid leukemia cell line HL-60 to explore a novel therapeutic target of leukemia.

METHODSHuman CaMK II N gene expression vector pcDNA3.1/hCaMKIIN or empty vector pcDNA3.1/myc-His (-) B was transfected into HL-60 cells by Lipofectamine 2000. Human CaMK II N proteins of transfected cells were detected by Western blot. Cell proliferation affected by human CaMKIIN was determined by MTT. Colony-forming assay was performed by soft agar growth system. The cells transfected with CaMKIIN were stained with Hoechst 33342 to detect the apoptotic proportion under fluorescence microscopy. Cell cycle was analyzed by flow cytometry.

RESULTSHuman CaMKIIN was stably transfected into HL-60 cells, and overexpression of human CaMKIIN inhibited the proliferation of HL-60/CaMKIIN cells compared to HL-60/mock cells and HL-60 cells [(0.44 ± 0.03) vs (0.94 ± 0.05) vs (0.94 ± 0.04), P<0.01]. The colony formation of HL-60/CaMKIIN was also markedly smaller[(21.00 ± 3.05)/500] than that of mock-transfected [(111.00±4.58)/500]] and control cells [(119.00±6.09)/500] (P<0.01). After 72 hrs-culture, the apoptotic proportion in cells transfected with CaMK II N was obviously higher than of cells transfected with mock DNA or control [(22.49 ± 2.15)% vs (7.17 ± 0.72)% vs (6.40 ± 0.55)%, P<0.01]. Up to (82.97 ± 2.90)% human CaMKIIN/HL-60 cells were arrested at G0/G1 phase, which was more than mock-transfected [(40.53 ± 2.38)%] and control cells [(41.63 ± 2.27)%] (P<0.05). Human CaMKIIN could down-regulate expression of Bcl-2 in transfected cells.

CONCLUSIONCaMK IIN up-regulation could inhibit proliferation and induce apoptosis of human acute myeloid leukemia cell HL-60.

Apoptosis ; Cell Proliferation ; Genetic Vectors ; HL-60 Cells ; Humans ; Proteins ; genetics ; metabolism ; Transfection ; Up-Regulation

OBJECTIVETo detect human bocavirus (HBoV) and investigate its genetic and evolutionary characteristics in children with acute respiratory infection in Tianjin, China.

METHODSA total of 1,259 samples of nasopharyngeal aspirates were collected from children with a confirmed diagnosis of acute respiratory infection between January and December, 2012. Viral nucleic acid was extracted, HBoV was detected by real-time quantitative PCR, and the gene segments of nucleocapsid protein of HBoV in positive samples were amplified by PCR. Several products were randomly selected and sequenced.The sequence obtained was compared with the known sequence of HBoV, and a phylogenetic analysis was performed. All the samples were examined to detect for other common respiratory tract viruses.

RESULTSAmong the 1,259 samples, the positive rate of HBoV was 4.53% (57/1,259), and among the 57 samples with positive HBoV, 75% (43/57) were positive in children with an age of 6-36 months. The positive rate of HBoV in children peaked in summer (from June to August), and there was a mixed infection with other viruses. Sequence analysis was performed for the PCR products from 36 positive samples, and the presence of HBoV was confirmed, with a higher homology to the known sequence of HBoV.

CONCLUSIONSIn Tianjin, acute respiratory infection in some children may be associated with HBoV infection, which is commonly seen in infants with an age of 6-36 months. The peak of HBoV infection occurs in summer. The phylogenetic analysis shows a high homology to the known sequence of HBoV, with few gene sequence variations.

Bocavirus ; classification ; isolation & purification ; Child, Hospitalized ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Phylogeny ; Polymerase Chain Reaction ; Respiratory Tract Infections ; virology ; Seasons

OBJECTIVETo screen the differentially expressing genes between silicotic lung tissue and normal lung tissue, to identify the differentially expressing genes of matrix metalloproteinase-12 (MMP-12) and Cathepsin E and to explore the roles of those genes in silicosis development.

METHODSThirty male SD rats were divided randomly into two groups: control group (6 rats) and exposure group (24 rats) which was exposed to SiO2 by intra-tracheal perfusion. On the 30 th, 60 th and 90 th days after exposure, 8 rats in model group and 2 rats in control group were executed and the lung tissues were obtained. The morphologic changes of lung tissues were observed with HE staining and VG staining under a light microscope. The gene microarrays were used to identify differentially expressing genes of lung tissues in rats exposed to SiO2 for 60 days. Two significantly up-regulated genes, MMP-12 and Cathepsin E, were validated using RT-PCR, immunohistochemistry and Western Blot assay.

RESULTSA total of 338 differentially expressing genes were identified from the 26 962 genes between silicotic rats and normal rats, including 267 up-regulated genes and 71 down-regulated genes. The results of RT-PCR showed that in the lung tissues of exposure group on the 30 th, 60 th and 90 th days, the mRNA expression levels of MMP-12 were 4.306, 5.338, 6.713 times higher than those in the control group, the mRNA expression levels of Cathepsin E were 1.434, 2.974, 3.889 times higher than those in the control group, respectively. The results of immunohistochemical showed that in the lung tissues of exposure group on the 30th, 60th and 90th days, the mRNA expression levels of MMP-12 were 1.435, 1.746, 2.069 times higher than those in the control group, the mRNA expression levels of Cathepsin E were 1.372, 1.663, 2.103 times higher than those in the control group, respectively. The results of immunohistochemical showed that in the lung tissues of exposure group on the 30th, 60th and 90th days, the expression levels of MMP-12 protein were 1.214, 1.531, 1.959 times higher than those in the control group, the expression levels of Cathepsin E protein were 1.262, 1.828, 1.907 times higher than those in the control group, respectively. Compared with the control group, the mRNA and protein expression levels of MMP-12 and Cathepsin E in lung tissues of exposure group were significantly up-regulated (P < 0.05).

CONCLUSIONThe differentially expressing genes in rat lung tissues screened by gene chip were validated, which suggested that a complex gene regulatory network may be contributed to occurrence of silicosis. MMP-12 and Cathepsin E genes may be involved in the development of silicotic pulmonary fibrosis by degrading the basement membrane of alveolar wall and participating in the immune response.

Animals ; Cathepsin E ; genetics ; metabolism ; Gene Expression ; Lung ; metabolism ; Male ; Matrix Metalloproteinase 12 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Silicosis ; genetics ; metabolism

OBJECTIVETo investigate the efficacy and safety of percutaneous radiofrequency perforation and valvuloplasty in infants with pulmonary atresia with intact ventricular septum (PA/IVS).

METHODSFour infants (body weight 4 - 10 kg) aged 11 months, 9 months, 12 days and 9 months old, respectively, were hospitalized for dyspnea and cyanosis. All patients had a continuous murmur in the left second intercostal space. Doppler echocardiogram showed membranous pulmonary atresia with intact ventricular septum. Right ventriculogram showed a tripartite right ventricle, vasiform infundibulum, and membranous pulmonary valve atresia without ventriculocoronary connections. Descending thoracic aortogram showed good-sized confluent pulmonary arteries being filled from a ductus arteriosus. All the patients were taken up for radiofrequency perforation followed by a balloon dilatation. A 6F Judkins right coronary guiding catheter was positioned in the right ventricular outflow tract and under the atretic pulmonary valve membrane. The radiofrequency perforation catheter along with coaxial injectable catheter was then passed through the right coronary guiding catheter, using it as the guide to the imperforate membrane. The proximal end of the radiofrequency perforation catheter was then connected to radiofrequency generator. After the cusps of pulmonary valve were perforated, the coaxial injectable catheter was moved into the main pulmonary artery. A tiny floppy-tipped coronary guidewire was then passed through the coaxial injectable catheter into the main pulmonary artery and directed through the patent ductus arteriosus into the descending thoracic aorta or directed into pulmonary arteriola. Thereafter, serial balloon dilation catheters were introduced across the pulmonary valve, and dilations were sequentially performed with increasing balloon diameters. The balloon was dilated until the concave of the balloons disappeared. The radiofrequency energy (5 to 8 W) was delivered for 2 to 5 seconds once, but commonly twice, to perforate the valves. After a predilation with a 3 mm x 20 mm to 5 mm x 20 mm balloon at 6 - 14 atm pressure, the valve was subsequently dilated with 10 mm x 30 mm to 14 mm x 30 mm balloon once or twice. The duration of procedures was 120 to 150 min and exposure time was 25.4 to 43.9 min.

RESULTSThe primary procedure was successful in all the infants except one who died early of cardiac perforation with tamponade. After a follow-up period ranging from 2 to 8 months (mean 4.3 m), the remaining 3 survivors achieved complete biventricular circulation. Two of them were awaiting occlusion of the patent ductus arteriosus and 1 needed right ventricular outflow tract reconstruction because of infundibular obstruction.

CONCLUSIONPA/IVS consists of 0.7% to 3.1% of congenital heart defects. 85% of the untreated patients die within half a year. Surgical repair for the infants with PA/IVS is associated with a high mortality. In carefully selected patients with PA/IVS, radiofrequency perforation and balloon dilatation of the pulmonary valve is feasible and may represent a new alternative to surgery due to its low mortality and avoidance of cardiopulmonary bypass.

Balloon Occlusion ; Catheter Ablation ; methods ; Catheterization ; methods ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Pulmonary Atresia ; physiopathology ; therapy ; Pulmonary Valve ; surgery ; Ventricular Septum

OBJECTIVETo investigate the effects of the self-developed anti-heparanase polypeptide antibodies on growth and invasion of human hepatocellular carcinoma HCCLM6 cells.

METHODSUsing MTT, flow cytometry, plate clone formation, transwell invasion and heparan degrading enzyme assay, the growth and invasion changes of human hepatocellular carcinoma HCCLM6 cells by co-culture with each of three self-developed rabbit anti-heparanase polyclonal antibodies were detected.

RESULTSCompared with normal rabbit IgG, in the presence of each anti-heparanase polypeptide antibody, the growth, cell cycle and clone formation remained unchanged, and under the P1 or P2 anti-heparanase polypeptide antibody (with final concentration 100 microg/ml), the cell invasiveness was inhibited by 52.5% and 36.6%, respectively, and the heparanase activity was inhibited by 42.9% and 39.1%, respectively.

CONCLUSIONThe P1 and P2 anti-heparanase polypeptide antibodies can effectively inhibit the invasion ability and heparanase activity of liver cancer HCCLM6 cells. However, All the three antibodies have no effects on its growth, cell cycle and clone formation.

Antibodies ; pharmacology ; Carcinoma, Hepatocellular ; enzymology ; pathology ; Cell Adhesion ; Cell Differentiation ; Cell Line, Tumor ; Cell Movement ; Coculture Techniques ; Enzyme Activation ; Glucuronidase ; immunology ; metabolism ; Humans ; Liver Neoplasms ; enzymology ; pathology ; Neoplasm Invasiveness

To investigate the relationship between interleukin-18 (IL-18) and human acute graft-versus-host disease (aGVHD), 26 patients undergoing allogeneic peripheral blood stem cell transplantation (allo-PBSCT) were included in this study. IL-18 secreted by peripheral blood mononuclear cells (MNCs) was measured by enzyme-linked immunosorbent assay (ELISA) before transplantation and during aGVHD. The results showed that grade I GVHD and grades III-IV GVHD developed in 10 and 5 cases, respectively. The levels in the supernatants of MNCs from patients with aGVHD were significantly higher than those in the cases without aGVHD. The levels of IL-18 were correlated with the severity of aGVHD. It is concluded that IL-18 plays an important role in the development of aGVHD.
Acute Disease ; Adolescent ; Adult ; Child ; Graft vs Host Disease ; etiology ; Hematopoiesis ; Humans ; Infection ; etiology ; Interleukin-18 ; physiology ; Leukemia ; therapy ; Peripheral Blood Stem Cell Transplantation ; adverse effects

Objective:To investigate the clinical efficacy of external fixators combined with anterolateral thigh musculocutaneous flap for treatment of Gustilo type IIIB/C open tibiofibular fractures.Methods:A retrospective case series study was conducted to analyze clinical data of 15 patients with Gustilo type IIIB/C open tibiofibular fractures admitted to Ruihua Hospital of Soochow University from March 2016 to June 2019. There were 11 males and 4 females, with the age of (48.5±12.6)years (range, 22-67 years). All patients underwent emergency debridement in stage I, the major blood vessels, nerves and tendons were inspected and repaired, and the fracture ends were fixed by external fixator. There were different degrees of wounds necrosis, infection and bone defect after operation. After debridement in stage II, the soft tissue defects with the dimension of 10.0 cm×5.0 cm to 30.0 cm×8.0 cm were repaired with anterolateral thigh musculocutaneous flaps whose areas ranged from 10.5 cm×5.5 cm to 30.5 cm×8.5 cm. All donor areas of the musculocutaneous flaps were sutured directly in stage I. The healing of the donor areas and musculocutaneous flaps were observed within 2 weeks after operation. At the last follow-up, the shape and sensory recovery of the flap, healing of fractures and related complications were observed. The lower extremity functional scale (LEFS) was used to evaluate the injured limb function.Results:All patients were followed-up for 12-32 months [(22.0±5.8)months]. All donor areas were healed by first intention, leaving only linear scars. The musculocutaneous flaps survived completely in all patients. Partial necrosis of large area of musculocutaneous flap occurred in 2 patients, and healed after debridement and skin grafting. Another patient had vascular crisis after musculocutaneous flap operation and survived after the embolized vein repaired by contralateral great saphenous. At the last follow-up, the shape of flap recovered well, and the feeling partially recovered with the two-point discrimination of 18-26 mm. All fractures healed well, and there were no serious infection-related complications such as osteomyelitis. The LEFS score was 47-69 points [(59.0±9.5)points].Conclusion:Theexternal fixator combined with anterolateral thigh musculocutaneous flaps for treatment of Gustilo type IIIB/Copen tibiofibular fractures can better restore the appearance of soft tissue defect of the lower leg, and can effectively reduce the occurrence of severe infection-related complications.