OBJECTIVETo establish fluorescent quantitative PCR method for detecting human herpes virus type 6 (HHV6).

METHODSAccording to the specific sequence of human herpes virus type 6 genes, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The fragment generated from HHV 6 gene as template was cloned into the pMD18-T vector which was constructed from the pUC 18. And the positive recombinant plasmid was 1:10 diluted and used as standard quantitative template to make the standard curve for sample detection.

RESULTSThe standard curve indicated the linear relationship between Ct (cycle threshold) and template concentration. The clinical samples from 135 cases were detected by this system, 16 cases among 135 were positive.

CONCLUSIONThe fluorescent quantitative PCR method for the detection of human herpes virus type 6 is simple and accurate, and this method may be helpful to clinical diagnosis.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; DNA Primers ; DNA Probes ; DNA, Viral ; genetics ; Female ; Herpesvirus 6, Human ; genetics ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Purpura, Thrombocytopenic, Idiopathic ; diagnosis ; virology ; Reproducibility of Results ; Roseolovirus Infections ; diagnosis ; virology ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo provide scientific basis for natural cultivation of Fritillaria cirrhosa.

METHODThe cluster analysis to the Bray--Curtis distance coefficient of community plot was carried out to divide the community types of F. cirrhosa.

RESULT AND CONCLUSIONThe community of F. cirrhosa's natural cultivation can be divided into six types: Sibiraea angustata community, Rhododendron litangense community, Salix sclerophylla community, Dasiphora fruticosa + Spiraea alpina + S. myrtilloides community, Sabina pingii var. wilsonii community, Polygonum viviparum + P. sphaerostachyum community, and the ecological characteristics such as the composition of community and ecological distribution have been showed.

Altitude ; China ; Climate ; Cluster Analysis ; Conservation of Natural Resources ; methods ; Ecosystem ; Fritillaria ; growth & development ; Pharmacognosy ; Plants, Medicinal ; growth & development ; Temperature

BACKGROUNDWe reviewed the outcomes of reoperations for 29 patients (30 limbs) who had undergone occluded arterial bypass in the lower limbs from May 1996 to September 2005.

METHODSThe 30 lower limbs of the 29 patients with arteriosclerotic obstruction received 44 reoperations, including thrombectomy alone (group T, 27) and inflow or outflow reconstruction plus thrombectomy (group C, 17). Among the 17 operations in group C, 17.6% (3/17) were inflow reconstructions involving the axillary-femoral (1), aorta-iliac (1) and aorta-femoral (1) arteries, and 76.4% (13/17) outflow reconstructions involving the femoral-popliteal bypass-tibial (8), femoral-tibial (1), femoral-popliteal bypass-popliteal arteries below the knee (2), and the femoral-popliteal bypass-tibial-peroneal trunk (2). One patient (1 limb) underwent both inflow and outflow reconstructions with an iliac arterial stent and a graft-popliteal anastomosis patch. Polytetrafluoroethylene (PTFE) grafts were used in the inflow or outflow reconstructions above the knee. Autovenous grafts or autovenously combined PTFE grafts were used in the outflow reconstructions below the knee.

RESULTSThe percentages of Fontaine stage III and IV before primary operation and reoperation were 60% (18/30) and 86.7% (26/30), respectively (P < 0.05). Four patients died of heart attack (2), stroke (1) and multiple organ failure (1) after reoperations. Among them, only 1 patient underwent occluded bypass, and others, patent bypass. Five patients after patent bypass are still alive. The accumulative patent rate was 28.6% (8/28). The average duration of patency in groups T and C was (4.16 +/- 5.68) (0.13 - 24) months and (7.14 +/- 6.37) (0.26 - 21) months, respectively (P > 0.05). Among 42 reoperations, 19 failed within 1 month in groups T (16) and C (3) (P < 0.01). Nine patients had limb amputated (10/28 limbs, 35.71%) because of graft infection (2 limbs), pseudo aneurysm at anastomosis (1 limb), and gangrene caused by failed grafts (7 limbs). The amputation was performed on 6 limbs within 1 month and on 4 limbs 1 month after reoperation (P > 0.05). The rate of limb salvage was 64.29% (18/28).

CONCLUSIONSThe percentages of Fontaine stage III and IV before reoperation may be much higher than those before primary operation. Thrombectomy plus inflow/outflow reconstruction creates patency better than thrombectomy alone for re-occluded bypass.

Aged ; Aged, 80 and over ; Arteriosclerosis ; surgery ; Female ; Humans ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Reoperation ; Thrombectomy

OBJECTIVETo review the follow-up results of the crural artery bypass.

METHODSSixty-five limbs in 64 patients with long stenosis or occlusion in femoral artery and popliteal artery were performed 65 times femoral-crural artery bypass surgery or femoral-popliteal-crural bypass surgery during April 2001 to July 2007. The ankle-brachial index before bypass surgery was 0.35 +/- 0.20 in anterior tibial artery and 0.38 +/- 0.21 in posterior tibial artery. Critical limb ischemia was 93.8%.

RESULTSThe ankle-brachial index after bypass surgery was 0.84 +/- 0.26 in anterior tibial artery and 0.83 +/- 0.22 in posterior tibial artery. The perioperative mortality rate was 1.6%, the perioperative amputation rate was 1.5%. Fifty-four patients 54 limbs were followed up. The average follow-up time was (24.1 +/- 16.6) months. The follow-up limb salvage rate was 85.2%. The follow-up mortality rate was 25.9%. Critical limb ischemia decreased as 13.0%. The follow-up ankle-brachial index was difference with before and after bypass surgery as 0.66 +/- 0.26 in anterior tibial artery and 0.64 +/- 0.25 in posterior tibial artery. It was no difference in cumulative limb salvage rate, cumulative primary and secondary patency rate by comparing autogenous vein with composite vascular as graft and comparing femoral-crural artery bypass surgery with femoral-popliteal-crural bypass surgery as surgical method.

CONCLUSIONSWhen the patients are failed in endovascular intervention or have long stenosis or occlusion in femoral artery and popliteal artery to face to amputation, the crural artery bypass is a feasible method. It's helpful to improve the secondary patency rate and limb salvage rate by enhancing the follow-up after operation and early intervention.

Adult ; Aged ; Aged, 80 and over ; Arterial Occlusive Diseases ; surgery ; Female ; Femoral Artery ; surgery ; Follow-Up Studies ; Humans ; Leg ; blood supply ; Male ; Middle Aged ; Popliteal Artery ; surgery ; Retrospective Studies ; Treatment Outcome ; Vascular Surgical Procedures

BACKGROUNDDyslipidemia, a well-known risk factor for cardiovascular disease, is common in patients with kidney disease. Recent studies discerned that dyslipidemias play a critical role in renal damage progression in renal diseases, but the association between dyslipidemias and chronic kidney disease (CKD) in the general population remains unknown. Thus, we assessed whether the growing prevalence of dyslipidemia could increase the risk of CKD.

METHODSA total of 4779 middle-aged and elderly participants participated in this study. Dyslipidemias were defined by the 2007 Guidelines in Chinese Adults. Incident CKD was defined as albuminuria and/or reduced estimated glomerular filtration rate (eGFR, < 60 ml×min(-1)×1.73 m(-2)). Regression analysis was used to evaluate the association between dyslipidemia and albuminuria/reduced eGFR.

RESULTSParticipants with hypercholesterolemia exhibited a greater prevalence of albuminuria and reduced eGFR (10.0% vs. 6.1%, P = 0.001; 4.0% vs. 2.4%, P = 0.028, respectively). Both hypercholesterolemia and low high density lipoprotein cholesterol (HDL-C) were independently associated with albuminuria (odds ratio (OR) 1.49; 95% confidence interval (CI) 1.08 - 2.07 and OR 1.53; 95%CI 1.13 - 2.09, respectively). The multivariable adjusted OR of reduced eGFR in participants with hypercholesterolemia was 1.65 (95%CI 1.03 - 2.65). As the number of dyslipidemia components increased, so did the OR of CKD: 0.87 (95%CI 0.65 - 1.15), 1.29 (95%CI, 0.83 - 2.01), and 7.87 (95%CI, 3.75 - 16.50) for albuminuria, and 0.38 (95%CI 0.21 - 0.69), 1.92 (95%CI 1.14 - 3.25), and 5.85 (95%CI 2.36 - 14.51) for reduced eGFR, respectively.

CONCLUSIONSOur findings indicate that dyslipidemias increase the risk of CKD in the middle-aged and elderly Chinese population. Hypercholesterolemia plays an important role in reducing total eGFR. Both low HDL-C and hypercholesterolemia are associated with an increased risk for albuminuria.

Aged ; Albuminuria ; epidemiology ; etiology ; physiopathology ; Cross-Sectional Studies ; Dyslipidemias ; complications ; epidemiology ; physiopathology ; Female ; Glomerular Filtration Rate ; physiology ; Humans ; Male ; Middle Aged ; Renal Insufficiency, Chronic ; epidemiology ; etiology ; physiopathology

OBJECTIVETo identify antiviral activity of Toddalia asiatica against influenza virus type A in vitro.

METHODMore than two hundred Chinese medicinal herb extracts were screened for antiviral activities against influenza A/PR/8/34 (H1N1) virus using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay for virus induced cytopathic effect (CPE) in a primary screening. Positive samples were picked up and were subjected to quantitative real-time polymerase chain reaction (PCR) to quantify reduction of H1N1 virus genomic RNA.

RESULTToddalia asiatica showed potent antiviral activities against H1N1 virus, with 50% effective concentration (EC50) value of 4.7 mg x L(-1) in MTS assay and 0.9 mg x L(-1) in quantitative PCR assay respectively. The cytotoxicity test of Toddalia asiatica generated a CC50 value of 187.2 mg x L(-1) and a selective index (SI) larger than 206 in quantitative PCR. Although the best antiviral activity of Toddalia asiatica was observed with co-treatment of influenza virus infection, it remained effective even when administrated 24 h before and after the initiation of infection.

CONCLUSIONThe results suggested that Toddalia asiatica compound extract could be a candidate for anti-H1N1 virus agent in the treatment of influenza.

Animals ; Antiviral Agents ; isolation & purification ; pharmacology ; Cells, Cultured ; Dogs ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; toxicity ; Influenza A Virus, H1N1 Subtype ; drug effects ; genetics ; Kidney ; cytology ; Plants, Medicinal ; chemistry ; RNA, Viral ; drug effects ; Rutaceae ; chemistry ; Time Factors

In order to prove the feasibility of preparation of the drug-incorporated stent by immersing stent wires in the monoclonal antibody (mAb) solution, fluorescence stain and image analysis were used to evaluate the L-PLA-coated stent. Absorption was measured using a radioisotope technique after preparing the mAb-incorporated stent, and the absorption curve was determined from the absorption data. In an in vitro perfusion circuit, the antibody was eluted from the stent matrices, and the related influence factors were evaluated based on the release data.
Absorption ; Alloys ; chemistry ; Antibodies, Monoclonal ; chemistry ; immunology ; Drug-Eluting Stents ; Graft Occlusion, Vascular ; immunology ; prevention & control ; Humans ; Lactic Acid ; chemistry ; Platelet Aggregation Inhibitors ; chemistry ; immunology ; Platelet Glycoprotein GPIIb-IIIa Complex ; immunology ; Polymers ; analysis ; chemistry ; Time Factors

Objective To evaluate the role of late course accelerated fractions.ted irradiation(LCAF) combined with concurrent chemotherapy in the management of esophageal carcinoma.Methods From March 1998 to July 2000,111 eligible patients were randomized into LCAF alone group(LCAF,57 patients)or LCAF plus concurrent chemotherapy group(LACF-CT,54 patients).The radiotherapy regimen was identical in the two groups,consisting of conventional fractionation in the first 2/3 course and accelerated fractionation in the second 1/ 3 course to a total dose of 68.4 Gy/41 Fx/44 d.Chemotherapy regimen consisted of four eycles of cisplatin 25 mg/ (m~2?d)plus fluorouracil 600 mg/(m~2?d)on day 1 to 3 every 4 weeks and was delivered on the first day of radiotherapy.Results The median follow-up time was 67.1 months(range 47.6-76.4 months).The 1-,3-,5- year survival rate was 67%,44% ,40% and 77%,39% 28% in LACF-CF and LEAF group,respectively(P =0.310).Grade 3+4 acute side-effact was 42% and 25% in LCAF-CT and LCAF group,respectively(P＜0. 05),with 3 treatment-related deaths in the LCAF-CT group.Conclusions Late course accelerated fractionated irradiation combined with concurrent chemotherapy has a trend towards improving the survival,at the cost of increasing acute side-effect.Its role needs further confirmation by larger sample studied in randomization.

Objective To study the changes in red blood cells(RBCs),hemoglobin(Hb),hematocrit(HCT),mean corpuscular volume(MCV),RBC distribution width variation coefficient(RDW-CV)and RBC distribution width standard deviation(RDW-SD)of leukocyte-poor red blood cells preserved in navy ship force on a long voyage.Methods According to the Requirement of Health Examination for Blood Donors(GB 18467 -2011),red blood cells in additive solution leukocytes reduced were randomly collected from ten healthy voluntary blood donors one day before sailing.Each blood sample was preserved.The relevant parameters were detected as data before sailing.Each blood sample was divided into two parts,one in test group and the other in control group.Samples were taken from the two groups of leukocyte-poor red blood cells for the sampling detection after 21 days of sailing respectively.Results ①After sailing, the concentrations of RBCs and Hb declined in both groups,and the difference of RBC concentrations was not significant(P=0.319),but that of the concentrations of Hb was significant(P=0.002).②After sailing, the concentrations of HCT and MCV increased, but the change of HCT was significant(P=0.015),while that of MCV was insignificant(P=0.051).③After sailing,the concentrations of RDW-SD and RDW-CV were higher,but the change of RDW-SD was significant(P<0.001),while that of RDW-CV was not(P=0.528).Conclusion When leukocyte-poor red blood cells are preserved in navy ship force,the concentrations of RBC and Hb decrease,while the concentrations of HCT,MCV,RDW-SD and RDW-CV increase with the prolongation of preservation.A long voyage has some impact on the parameters of red blood cells in additive solution leukocytes reduced,as is the case with the conventional blood storage refrigerator(4 ±2)℃.

Objective To explore the influence on and changes in the biochemical indices of red blood cells(RBCs)in additive solution leukocytes reduced preserved in navy ship force on a long voyage.Methods According to the Requirement of Health Examination for Blood Donors(GB 18467 -2011),RBCs in additive solution leukocytes reduced were prepared from 10 healthy voluntary blood donators one day before sailing.Each blood sample was divided into two parts,one in test group and another in control group.All the groups had samples taken for the biochemical index detection after 1,3,5,7,14 and 21 days of sailing respectively.Results ①The change in total protein(P=0.235)and albumin (P=0.119)concentration was not obvious,and the difference between the two groups was not significant.②The change in total creatinine(P=0.001)and uric acid(P=0.001)concentration was obvious, but the difference between the two groups was not significant.③The change in total cholesterol(P=0.354)concentration was not obvious,but the change in triglycerides(P=0.005)concentration was significant.The difference between the two groups was not significant.④The concentration of lactate dehydrogenase, hydroxybutyrate dehydrogenase and creatine kinase increased with the time of preservation(P<0.001).The difference between the two groups was not significant.⑤The interaction between grouping effect and time effect had no significant influence on the concentration of osmolarity(OSM)(P=0.968)and glucose(Glu) (P=0.406).Between the two groups,the difference of concentrations of OSM(P=0.569)and Glu(P=0.115)was not significant.Conclusion Under the 4 class sea conditions, a long voyage has some impact on the storage of RBCs in additive solution leukocytes reduced,as in the conventional blood storage refrigerator(4 ±2)℃.The results of this study have important clinical implications for our further study of marine blood support.