BACKGROUND: It is necessary to establish a high throughput screening system for anti -inflammatory drugs for rheumatoid arthritis.OBJECTIVE: To construct an eukaryotic expression vector p4CCL20-ZsGreen1-DR with the NF-kB cis-acting element 4×CCL20motif as an enhancer, SV40 as a promoter, and ZsGreen1-DR as a reporter gene.METHODS: The target fragment SV40 was PCR amplified using PGL2-control plasmid as a template. KpnⅠ/Bam HⅠ restriction sites were introduced into the flank of the target fragment. Then, pSV40-ZsGreen1-DR vector was constructed by cloning the target fragment into pZsGreen1-DR plasmid. Finally, p4CCL20-ZsGreen1-DR plasmid was constructed by cloning the double strand DNA of 4×CCL20 motif (with BglⅡ and EcoRⅠ sticky ends at the 5’ and 3’ terminus, respectively) into the corresponding restriction sites of pSV40-ZsGreen1-DR vector (upstream of SV40 promoter).RESULTS AND CONCLUSION: DNA sequencing demonstrated successful construction of p4CCL20-ZsGreen1-DR plasmid.The construction of p4CCL20-ZsGreeR plasmid might be useful to establish a high throughput screening system for anti -inflammatory drugs.

Objective To investigate the clinical value of spinal nonfusion interspinous Coflex system for the degenerative lumbar spinal stenosis.Methods From March 2008 to August 2009,26 patients with degenerative lumbar spinal stenosis were treated by decompression with posterior spinous process and interspinous implant Coflex folder method,including 11 males and 15 females,with the mean age of 65.4years(range,45-78 years).L3,4 segment in 7 cases,L4,5 segment in 13 cases,concomitant L3,4,L4,5 segment stenosis in 6 cases.Patients were scanned by MRI and CT confirmed L3,4 and(or)L4,5 segments of the ligamentum flavum thickening,proliferation of small-joint bone,merger disc herniation causing central tube and lateral fossae stricture,nerve root or coccygeal plexus compression.Lumbar anteroposterior,lateral and flexion-extension X-ray films preoperatively,and at follow-up were used to measure the following parameters by eFilm and CAD software:the anterior and posterior disc space height,the range of motion at surgical level,and the segment of the spinal canal area of responsibility.Postoperative standard Japanese Orthopaedic Association(JOA)score for preoperative and postoperative scores.Results Patients were followed up 12-24months.The pain relieved in all patients.The JOA scores improved from(15.46 ±4.30)preoperatively to (24.50±1.58)postoperatively,the vertebral canal area of the affected segment was(218.4±16.2)mm2 before the operation,(264.6±9.9)mm2 after the operation.Single segment anterior disc space height did not change significantly,the difference was not statistically significant.The disc space posterior height increased compared with the preoperative height,and gradually decreased with time.No patients suffered Coflex loosening,fracture and emerge.Conclusion The treatment of lumbar spinal stenosis with Coflex system has satisfactory effect in minimal invasiveness and high security,which provides a safe and effective therapy for degenerative lumbar spinal stenosis.

OBJECTIVE:To study the preparation technique of dipyridamole microcapsules METHODS:Using the method of orthogonal design,the preparation technique of dipyridamole microcapsules was optimized RESULTS:The results showed that with the simple agglutination method,the encapsulation rate was significantly related to the ratio of coating material to dipyridamole of preparation(P

OBJECTIVETo determine the factors affecting post-operative velopharyngeal function of cleft palate patients aged over two years old.

METHODSThe data on 245 patients, including first visit case records and those in the database, were searched and recorded. Post-operative velopharyngeal function and clinical features, such as cleft ratio, adequate ratio, diagnosis, operative age and method, were analyzed retrospectively.

RESULTSCleft ratio and adequate ratio in the velopharyngeal competence (VPC) group were not statistically different from those in the velopharyngeal incompetence (VPI) group. The difference in VPC rate in different age groups and diagnosis was not significant. The post-operative VPC rates under different methods were significantly different.

CONCLUSIONThe current therapeutic schedule for patients younger than two years old is inappropriate for patients older than two years old. Cleft ratio is not the main factor affecting postoperative velopharygeal function. Sommerlad-Levator Veli Palatini reconstructional operation combined with sphincter pharyngoplasty can obviously improve the velopharyngeal function of patients aged 2 years old and up. This method should be applied in primary palatoplasty.

Child, Preschool ; Cleft Palate ; Female ; Humans ; Infant ; Male ; Palatal Muscles ; Pharynx ; Postoperative Period ; Reconstructive Surgical Procedures ; Retrospective Studies ; Velopharyngeal Insufficiency

OBJECTIVETo obtain the exposure-response functions that could be used in health-based risk assessment of particulate air pollution in China.

METHODSMeta analysis was conducted on the literatures on air particulate matter and its adverse health outcomes in China and worldwide.

RESULTSFor each health outcome from morbidity to mortality changes, the relative risks were estimated when the concentration of air particulate matter increased to some certain units.

CONCLUSIONThe exposure-response functions recommended here can be further applied to health risk assessment of air particulate matter in China.

Acute Disease ; Adult ; Air Pollutants ; adverse effects ; toxicity ; Asthma ; epidemiology ; etiology ; Bronchitis ; epidemiology ; etiology ; Bronchitis, Chronic ; epidemiology ; etiology ; Cardiovascular Diseases ; epidemiology ; etiology ; Child ; China ; Dust ; Environmental Exposure ; Hospitalization ; Humans ; Mortality ; Particle Size ; Risk ; Risk Assessment

Objective To explore the value of retrograde recanalization technique in treatment of TASCⅡ C/D femoropopliteal arteriosclerosis obliterans.Methods Totally 36 patients with TASCⅡ C/D femoropopliteal arteriosclerosis obliterans were retrospectively analyzed.Preoperative and postoperative ankle brachial index (ABI) and intermittent claudication were compared,as well as postoperative vascular patency rate and complications were analyzed.Results The retrograde recanalization technique were successfully performed in all 36 patients.The walking distance and ABI of 7 days,and 3,6,12 months postoperation were increased significantly compared with preoperation (all P＜0.05).The postoperative vascular patency rate of 3,6 and 12 months was 97.22％,91.67％ and 72.22％,respectively.There was no death nor amputation during the follow-up period,and the incidence of perioperative complications was 11.11 ％ (4/36).Conclusion Retrograde recanalization technique can be used to treat TASC Ⅱ C/D femoropopliteal arteriosclerosis obliterans when anterograde access failed.

OBJECTIVETo investigate human cytomegalovirus (HCMV) glycoprotein B (gB) genotypes and clinical features in neonates with congenital infections.

METHODSUrine samples were obtained from 67 neonates with HCMV infection confirmed by polymerase chain reaction (PCR). The gB gene fragment was amplified by nested PCR. HCMV gB genotyping was detected by restriction fragment length polymorphism.

RESULTSIn all these cases, the most prevalent genotype was gBl (50.7%), followed by gB3 (23.9%), gB2 (17.9%), and gBl/gB3 coinfection (7.5%); gB4 was not found. Moreover, gB1 was more prevalent in infants with liver damage (27/37, 73.0%) than in other symptomatic infants without liver damage (13/30, 43.3%; P < 0.05).

CONCLUSIONThe gBI genotype is the most prevalent in infants with congenital symptomatic HCMV disease, especially in those with liver damage, followed by genotypes gB3, gB2, and gB4.

Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; congenital ; urine ; virology ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Infant, Newborn, Diseases ; virology ; Male ; Viral Envelope Proteins ; genetics ; urine

OBJECTIVETo evaluate the expression of vascular endothelial growth factor C (VEGF-C) and peroxisome proliferators-activated receptors (PPARgamma) in extrahepatic cholangioadenocarcinoma (EHCAC) and to elucidate its correlation with clinicopathological factors and their significance in prognosis.

METHODSThe expressions of PPARgamma and VEGF-C were detected by immunohistochemistry in 69 cases of EHCAC, 12 cases of non-tumor bile duct epithelium, and their relationship to clinicopathological parameters and follow-up were analyzed.

RESULTSThe positive rate of PPARgamma expression in 69 cases of EHCAC was 59.4%, significantly higher than that in 12 cases of non-tumor bile duct epithelium (0%), (P < 0.01). The positive rate of VEGF-C in 69 cases of EHCAC was 84.1%, also significantly higher than 16.7% in 12 cases of benign bile duct epithelium (P < 0.05). PPARgamma expression was associated with clinical TNM stage and lymph node metastasis. VEGF-C expression was associated with lymph node metastasis. Cox analysis results showed that portal vein and/or hepatic artery invasion, lymph node metastasis and VEGF-C expression were independent prognostic factors of EHCAC (P < 0.05).

CONCLUSIONPPARgamma expression may play an important role during tumorigenesis of extrahepatic cholangioadenocarcinoma. The expressions of PPARgamma and VEGF-C are significantly correlated with the clinicopathological characteristics and biological behavior of EHCAC. Expression of VEGF-C is an independent prognosis factors in EHCAC. The detection of PPARgamma and VEGF-C is valuable for evaluation of prognosis of EHCAC.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; Cholangiocarcinoma ; metabolism ; pathology ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; PPAR gamma ; metabolism ; Proportional Hazards Models ; Survival Rate ; Vascular Endothelial Growth Factor C ; metabolism

AIMTo investigate the effect of resveratrol on EMMPRIN expression of macrophages.

METHODSHuman monocytic cell line THP-1 cells were co-cultured with EMMPRIN-highly-expressed MCF-7 cells; MMP-9 production was assayed by zymography. THP-1 cells were induced by PMA, expression of EMMPRIN was assayed by Western blotting. Cells were treated with resveratrol or PPARgamma agonist--pioglitazone during differentiation, EMMPRIN expression and MMP-9 activity were assayed. U937 cells were co-transfected with PPARy expression and luciferase-coding reporter vector, then cultured with pioglitazone or resveratrol, the activating capability of resveratrol on PPARgamma was evaluated by measuring the luciferase activity. THP-1 cells were pretreated with PPARgamma antagonist--GW9662 before pioglitazone or resveratrol treatment, then assayed for EMMPRIN expression and MMP-9 production.

RESULTSEMMPRIN expression was greatly increased during the differentiation from monocytes to macrophages; co-culturing with MCF-7 cells significantly increased MMP-9 production by monocytes. Both resveratrol and pioglitazone markedly inhibited EMMPRIN expression during monocytes differentiation. Resveratrol significantly activated PPARgamma and GW9662 greatly decreased the effect of resveratrol on EMMPRIN and MMP-9.

CONCLUSIONEMMPRIN expression is greatly up-regulated from monocytes to macrophages, which may play a role in inducing MMPs production by monocytes/macrophages. Resveratrol can significantly inhibit EMMPRIN expression via activating PPARgamma, which may be the underlying mechanism of its inhibitory effect on MMPs production by monocytes/macrophages.

Anilides ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Basigin ; biosynthesis ; genetics ; Blotting, Western ; Breast Neoplasms ; metabolism ; pathology ; Cell Differentiation ; drug effects ; Cell Line ; Cell Line, Tumor ; Coculture Techniques ; Dose-Response Relationship, Drug ; Female ; Humans ; Luciferases ; genetics ; metabolism ; Macrophages ; cytology ; drug effects ; metabolism ; Matrix Metalloproteinase 9 ; biosynthesis ; Monocytes ; cytology ; drug effects ; metabolism ; PPAR gamma ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Stilbenes ; pharmacology ; Thiazolidinediones ; pharmacology ; U937 Cells

3D Scaffolds with controlled porous structure were designed and fabricated by utilizing CAD and rapid prototyping techniques. A flow perfusion bioreactor, which allowed exposure of the culture cells to low levels of mechanical stimulation by fluid flow-induced shear stress, was developed in our lab. The scaffolds were pre-designed and the negative images of the designs were used to build the molds on a stereolithography (SL) apparatus with epoxy resins. Calcium phosphate cement paste was cast into the molds. And after pyrolysis, the 3D scaffolds with controlled internal pore architectures were obtained. Rabbit osteoblasts were seeded in 3D porous scaffolds, cultured in the flow perfusion bioreactor with media flow rate set at 2 mL/min and 6-well plates. At 3, 7, and 14 days, scanning microscopic evaluation showed excellent growth on the surface of scaffolds and poor viability of cells within microchannels in static cultures. In flow perfusion bioreactor, there was greater cellularity throughout the scaffolds and abundant deposition of extracellular matrix. Cells were also seen throughout the internal microchannels of scaffolds. These results represent that better mass transport of oxygen and nutrient occurred in the flow perfusion bioreactor and cells distribution in 3D porous scaffolds was improved.
Animals ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Cell Division ; Cells, Cultured ; Osteoblasts ; cytology ; Porosity ; Rabbits ; Skull ; cytology ; Tissue Engineering ; methods ; Tissue Scaffolds