Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

Objective To evaluate the clinical efficacy of multiple stents placement in the management of hilar cholangiocarcinoma,especially in the complex cases of which the hepatic ducts are invaded.Methods Forty-five consecutive patients with hilar cholangiocarcinoma were treated with percutaneous transhepatic placement of two or three self-expandable metallic endoprostheses.The cause of hilar obstructions in these patients were all cholangiocarcinoma,including Bismuth classification type Ⅱ(n 12 ),Ⅲa(n 17),Ⅲb(n 10),and Ⅳ(n 6).Two or 3 stents were placed in the configuration of T,Y or X over the strictures.Results Stent placement with 2 or 3 endoprostheses was successful in all patients.All patients showed significant decrease in serum bilirubin level.The mortality rate within 30 days of stent placement was 2.2%(1/45).The mean survival and stent patency times were 215.3 d(26— 516 d)and 181.5 d(26—473 d),respectively.Conclusion Deploying of multiple metallic stents is an effective method to treat complex hilar cholangiocarcinoma,especially for the cases of which hepatic ducts are invaded:the henatic ducts should be drained as much as nossible.

BACKGROUND: Studies have shown that titanium has good biological properties and regulate osteoblast adhesion, proliferation and differentiation, but there are few reports on the effect of titanium materials on bone defect repair and on immune regulation in rats. OBJECTIVE: To investigate the effect of titanium materials in the repair of osteoporotic bone defect in rats and its effect on immune regulation. METHODS: Bilateral ovaries were removed from 30 Wistar rats to establish osteoporosis models. Three months after modeling, a percutaneous bone defect with the diameter of 2.5 mm was prepared on the distal femur of all the 30 rats. These model rats were then randomly divided into three groups with 10 rats in each group: negative control group without any intervention, hydroxyapatite group filled with nano-hydroxyapatite, and titanium material group filled with titanium material. Serum interleukin-1, tumor necrosis factor-α and interleukin-6 levels were measured by enzyme-linked immunosorbent assay after 5 days of intervention, and meanwhile, serum T-lymphocyte subsets were detected by flow cytometry. After 5 and 10 weeks of intervention, hematoxylin-eosin staining was used for morphological observation of the defect site. RESULTS AND CONCLUSION: (1) After 5 days of intervention, the levels of interleukin-1, tumor necrosis factor-α and interleukin-6 in the titanium material group were significantly lower than those in the negative control and hydroxyapatite groups (P < 0.05). (2) After 5 days of intervention, the levels of CD4+and CD8+and the ratios of CD4+/CD8+in the titanium material group were significantly higher than those in the hydroxyapatite and negative control groups (P < 0.05). (3) Hematoxylin-eosin staining results showed that a small amount of bone tissues filled with a large amount of fibrous tissues and adipocytes formed in the negative control group at 5 and 10 weeks after intervention. A little bone formation was observed in the hydroxyapatite group at 5 weeks after intervention, and the implant began to degrade at 10 weeks and was gradually replaced by new bone tissues. A small amount of trabecular bones enriched with blood vessels arranged orderly in the titanium material group at 5 weeks after intervention; and at 10 weeks, the number of trabecular bone increased and the defect was basically healed. Taken together, the titanium plate can achieve good results in the repair of osteoporotic bone defects in rats, and moreover, it can control the immune response during the bone defect repair.

OBJECTIVETo investigate the oxidative damage to lung tissue and peripherial blood in PM2.5-treated rats.

METHODSPM2.5 samples were collected using an auto-sampling instrument in summer and winter. Treated samples were endotracheally instilled into rats. Activity of reduced glutathione peroxidase (GSH-Px) and concentration of malondialdehyde (MDA) were used as oxidative damage biomarkers of lung tissue and peripheral blood detected with the biochemical method. DNA migration length (microm) and rate of tail were used as DNA damage biomarkers of lung tissue and peripheral blood detected with the biochemical method.

RESULTSThe activity of GSH-Px and the concentration of MDA in lung tissue significantly decreased after exposure to PM2.5 for 7-14 days. In peripheral blood, the concentration of MDA decreased, but the activity of GSH-Px increased 7 and 14 days after experiments. The two indicators had a dose-effect relation and similar changing tendency in lung tissue and peripheral blood. The DNA migration length (microm) and rate of tail in lung tissue and peripheral blood significantly increased 7 and 14 days after exposure to PM2.5. The two indicators had a dose-effect relation and similar changing tendency in lung tissue and peripheral blood.

CONCLUSIONPM2.5 has a definite oxidative effect on lung tissue and peripheral blood. The activity of GSH-Px and the concentration of MDA are valuable biomarkers of oxidative lung tissue damage induced by PM2.5. The DNA migration length (microm) and rate of tail are simple and valuable biomarkers of PM2.5-induced DNA damage in lung tissues and peripheral blood. The degree of DNA damage in peripheral blood can predict the degree of DNA damage in lung tissue.

Animals ; DNA Damage ; drug effects ; Drug Administration Routes ; Drug Administration Schedule ; Lung ; drug effects ; pathology ; Lung Diseases ; blood ; chemically induced ; pathology ; Male ; Oxidative Stress ; Particle Size ; Particulate Matter ; administration & dosage ; toxicity ; Rats ; Rats, Wistar ; Seasons

Objective To find out the status of brick-tea type fluorosis in the epidemic areas.Methods Based on "Scheme for Epidemiological Brick-tea Type Fluorosis in Sichuan Province",ten counties were selected in Sichuan brick-tea areas and ten towns were selected in every county,then the epidemicologic survey was performed in children of 8～12 year-old and adults aged above 20 years old.Results 5044 children and 4053 adults were selected from brick-tea areas.The rates of dental fluorosis in children and adults were 55.69%(2809/5044)and 60.41%(4053/6709)respectively.The dental fluorosis was mainly of mild damage.The skeletal fluorosis found in X-ray film was 44.64%(167/1241)and in clinical examination,38.94%(3883/9973).The levels of urine fluoride in children and adults were 1.88 and 2.78 mg/L.The level of urine fluoride was not differenet among children of different age,but in adults it was higher in the elder than the younger.The level of fluoride in urine was related to the severeness of skeletal fluorosis(r=0.74).The detective rates of skeletal fluorosis in agricuIture,pasturing,and agriculture-pasturing areaswere 31.70%(1369/4318),50.04%(1228/2454),and 40.17%(1286/3201),respectively.The X-ray detecting rates of skeletal fluorosis in men and wonlen were 49.57%(229/462)and 41.72%(325/779) respectively(χ2=11.72,P＜0.05).Conclusion The prevalence of brick-tea type fluorosis is very serious in the regions studied.

OBJECTIVETo evaluate the technical feasibility of juxtarenal abdominal aortic aneurysm (AAA) repair with fenestrated endovascular stent-graft

METHODSA 64-year-old male was diagnosed with juxtarenal AAA with severe coronary artery stenosis, fenestrations was customized according to precise helical CT data to accommodate visceral and renal arteries. Under general anesthesia and dynamic supervision of digital subtraction angiography (DSA), juxtarenal AAA was excluded with the customized fenestrated stent-graft and balloon expandable mini stent-grafts were deployed into bilateral renal arteries respectively.

RESULTSAfter operation, DSA showed the patency of the super mesenteric artery, bilateral renal arteries and left hypogastric artery, no endoleak was found. The serum creatinine decreased slightly after operation. CT angiography revealed favorable morphology of the stent-graft without tortuosity, migration, disjoint and endoleak 10 days after the operation and patency of super mesenteric artery, bilateral renal arteries and left hypogastric artery.

CONCLUSIONSThe placement of customized fenestrated endovascular stent-graft is a feasible option for juxtarenal AAA.

Aortic Aneurysm, Abdominal ; surgery ; Blood Vessel Prosthesis Implantation ; instrumentation ; methods ; Feasibility Studies ; Humans ; Male ; Middle Aged ; Stents

Objective To investigate the impact of the regional cooperative chest pain center (CPC) on therapeutic time and short term outcome after primary percutaneous coronary intervention (PCI) of patients with ST segment elevated myocardial infarction.Methods 372 patients with ST segment elevated myocardial infarction were enrolled in the study who had received were operated primary PCI 18 months before and after the regional cooperative CPC was set up.There were 156 patients in the green channel group before the setup of CPC and 216 patients in the CPC group.Total ischemia time,first medical contact (FMC) time,FML-to-balloon (FMC2B) time,door-to-balloon (D2B) time,hospital mortality rates,cardiac failure rates on the next day after PCI,length of CCU stays and hospital stays were compared between the two groups.Results Compared to the green channel group,total ischemia time[(281.0±102.7)min vs.(365.2±115.6)min,P＜0.05],FMC time [(174.3±97.5) min vs.(225.4±104.6) min,P＜0.05],FMC2B time [(106.7±61.2) min vs.(139.8±75.7) min,P＜0.05] and D2B time [(75.2±45.4) min vs.(102.4±53.7) min,P＜0.05] of the CPC group were significant shorter.The rates of reaching the standard of FMC2B time (70.83％ vs.34.62％,P＜0.001) and D2B time (75.93％ vs.40.38％,P＜0.001)were significantly higher in the CPC group.Cardiac failure rates on the next day after PCI was lower in the CPC group (14.35％ vs.23.72％,P=0.021),and CCU stays was shorter [(64.3±13.72)h vs.(92.6±15.65)h,P=0.043].Conclusions Establishment of a standardized regional cooperative CPC requires combination and consideration of the characteristics of local resources FMC2B time and D2B time of STEMI patients can be shorten by a standardized CPC lending to further shortening of total ischemia time and improvement in cardiac function.