Objective:To investigate a potential role of Toll-like receptor 4(TLR4) ,a pathogen pattern recognition receptor, in the early phase of severely burned rats. Methods: Rats burn model(30% of total body surface area[TBSA],Ⅲgrade) were established with vapor at 108 C for 8 seconds. Rats were sacrificed before and 2,5,12,24,48 and 72 h after burning, and the spleen specimens and peripheral blood samples were harvested. TLR4 mRNA and TNF-?mRNA expression in splenocyte was measured by reverse-transcription PCR(RT-PCR); the expression of TLR4 protein were measured by Western bloting; the endothelial toxicity concentration in plasma was detected by limulus lysate test. Results: It was found that the expression of TLR4 mRNA.TNF-?mRNA,TLR4 protein,and the level of ET were significantly increased in burned group compared with normal control group. The expression of TLR4 mRNA and protein peaked at 8 h after burning, the expession of TNF-?mRNA peaked at 12 h.and the level of ET peaked at 8 h after burnings the peak values of them were (3. 66?0. 51),(2. 27?0. 19), (1.65?0. 23),and (11. 68?2. 63) Eu/ml, respectively, all significantly higher than those of the control group(P

Objective To establish a stable animal model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were replicated. After immediate subcutaneous suture,the mice were randomly divided into four groups,ie,Group A was inoculated with 50 μl sterile PBS solution),Groups B,C and D were inoculated with 50 μl suspension containing 1 × 106,1 × 108 and 1 × 1010 colony forming unit (CFU)/ml bioluminescent methicillin-resistant staphylococcus aureus (MRSA) respectively.Then,the diet behavior of each group was observed and the mean weight and mortality of each group were also recorded at different time points.The bioluminescent intensity of fluoresce in the wounds was recorded at different time points by using the charge-coupled device (CCD) based imaging system.Local wound tissues were incised at 24 hours after inoculation for HE staining so as to observe wound inflammatory reaction in each group.Wound healing time of each group was also recorded. Results ( 1 ) Average weight:Groups A and B showed unobvious changes in weight; Group C lightened until day 3 after inoculation and then recovered gradually to the preinoculation level at day 14; Group D lightened gradually until death.(2)Mortality:Groups A and B had no death; Group C had 10％ deaths at day 14; Group D had 100％ deaths.(3) Bioluminescent intensity of wounds:Groups A and B showed a gradual weakened luminescence since the day of inoculation and had almost complete disappearance at days 5 and 7 respectively; there was no sign of obvious increase or decrease in Group C from the day of inoculation till day 14 ; Group D had a gradual increase since the day of inoculation and the luminous area expanded until the death.(4) HE staining at 24 hours after inoculation:all the four groups showed inflammatory cell infiltration,especially in Groups C and D.(5) Wound healing time:wound healed at days 5 and 7 after inoculation in Groups A and B; the wounds showed no healing even at day 14 in the Group C,but the wounds length and area did not show obvious enlargement or diminishment ; the wounds extended gradually until the death in the Group D,since the day of inoculation. Conclusions The inoculation of 50 μl suspension with 1 × 108 CFU/ml bioluminescent MRSA to full-thickness skin incision rats allows direct,real-time dynamic and continuous detection of the occurrence and development of the wound infections.The infection model is easy to make and has stability and high repeatability.

Objective To investigate the expression of circulating microRNA (miRNA) of rats with hypobaric hypoxia‐induced pulmonary hypertension (HPH) .Methods Commercial rat miRNA microarray was employed to detect and analyze the circulating miRNA profile in the serum samples of Sprague‐Dawley rats with hypobaric hypoxia‐induced HPH and controls .Furthermore ,differentially expressed candidate circulating miRNAs between HPH and control groups were validated by Real‐time quantitative PCR based on the case‐control study ,and receiver operating characteristic curve (ROC ) analysis was used to test the performance of four differentially expressed circulating miRNAs in discriminating HPH and control groups .Results Compared with those in the control group ,13 upregulated miRNAs and 10 downregulated miRNAs were identified in hypobaric hypoxia‐induced HPH rats by using miRNA microarray . And differentially expressed miR‐451 , miR‐505 , let‐7d and miR‐214 were validated by using RT‐PCR .ROC analysis showed that the area under the curve of miR‐451 ,miR‐505 and let‐7d was 0 .979 ,0 .938 and 0 .993 in discriminating HPH and control groups ,respectively .Conclusion The aberrant expression of circulating miR‐451 ,miR‐505 and let‐7d in serum may be correlated with the pathogenesis of HPH .

Objective To explore the effect and safety of transcranial approach for spheno-orhital meningioma. Design Retro- spective case series. Participants Thirty-two patients being operated with transcranial approach. Twenty-four cases were meningothelial meningiomas, 3 cases were fibrous meningiomas, 1 case was psammomatous meningioma, 2 cases were atypital meningiomas, 2 case were malignant meningiomas. Methods All patients underwent frontal-temporal craniotomy, the involved sphenoid wing bone and peri- orbit were removed to prevent recurrence. The superior orbital fissure and optic canal were decompressed, the dural and periorbital de- feet were repaired by autogenous temporal fascia or artificial dura. Main Outcome Measures Preoperative and postoperative exoph- thalmus and eyeball movement, the extent of tumor resection, the ratio of recurrence. Results The extent of tumor resection: 8 cases were Simpson gradeⅡ, 20 cases Simpson gradeⅢ, 4 cases Simpson grade IV. After surgery, proptosis were improved in all patients, ophthalmoplegia was found in 6 eases. There was no operation-related death or other significant complication. Tumor recurred in 6 cas- es. Conclusions Adequate exposure of the tumor and bony decompression of the cranial nerves can result from transcranial approach, all the involved bone should be removed in order to prevent recurrence. This approach is relatively safe and the ptoptosis are improved significantly. Complete surgical resection is difficult because of the involvement of the orbital apex, superior orbital fissure and cav- ernous sinus.

Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Infant ; Male ; Thrombasthenia ; classification ; diagnosis

OBJECTIVETo explore the feasibility of replacement of ordinary syringes by auto-disposal syringes (AD) in China.

METHODSSampling survey was conducted in Hebei province to collect data on the direct medical spending of hepatitis B and their Disability Adjusted Life Year (DALY) losses. The financial burden on hepatitis B was calculated based on the result of field survey as well as through data-base of the Second National Health Services Survey of MOH in 1998. The cost-benefit analysis was employed, in which the saving of the financial burden on hepatitis B patients attributing to unsafe injections was taken as the benefit of intervention and the increased expenditures of AD syringes as the costs.

RESULTSThe total financial disease burden of Hepatitis B in China in 2001 was between RMB 16.19 and 20.17 billion Yuan and the financial disease burden per patient was between RMB 1755.38 and 1409.64 Yuan. Study on the cost-benefit analysis in the replacement intervention program showed high financial feasibility. The Benefit-Cost Ratio (BCR) of the measure was between 32.46 and 97.39, which was bigger in rural areas than that in cities. The direct BCR was less than one when the price gap between ordinary syringe and AD exceeded RMB 0.24 Yuan.

CONCLUSIONThe intervention of safe injection practice through universal usage of AD to replace ordinary syringe seemed to be applicable due to its high potential economic benefits.

China ; epidemiology ; Cost-Benefit Analysis ; Cross Infection ; economics ; etiology ; prevention & control ; Cross-Sectional Studies ; Disposable Equipment ; economics ; statistics & numerical data ; Equipment Reuse ; statistics & numerical data ; Female ; Hepatitis B ; economics ; transmission ; Humans ; Injections ; adverse effects ; economics ; instrumentation ; Male ; Risk Factors ; Sampling Studies ; Surveys and Questionnaires ; Syringes ; economics

ATP-NAD kinase phosphorylates NAD to produce NADP by using ATP, whereas ATP-NADH kinase phosphorylates both NAD and NADH. Three NAD kinase homologues, namely, Utr1p, Pos5p and Utr1p, exist in the yeast Saccharomyces cerevisiae, which were all confirmed as ATP-NADH kinases and found to be important to supply NADP(H) for yeast cells. In S. cerevisiae, fatty acid beta-oxidation is restricted to peroxisomes and peroxisomal NADPH is required for beta-oxidation of unsaturated fatty acids with the double bonds at even positions. Single and double gene disruption strains of NAD kinase genes, i.e., utr1, pos5, yef1, utr1yef1, utr1pos5 and yef1pos5 were constructed by PCR-targeting method. The utilization ability of these mutants for unsaturated fatty acids with the double bonds at even or uneven positions was examined, with wild type BY4742 as positive control cell, and fatty-acyl-CoA oxidase gene deletion mutant (fox1) and peroxisomal NADP-dependent isocitrate dehydrogenase isoenzymes gene deletion mutant (idp3) as negative control cells. The results indicated that the NAD kinase homologues, especially Pos5p, were critical for supplying NADP and then NADPH in peroxisomal matrix. NADP, which was supplied mainly by Utr1p, Pos5p and Yef1p, particularly by Pos5p, was proposed to be able to transfer from outside of peroxisome into peroxisomal matrix and then converted to NADPH by Idp3p.
Fatty Acids, Unsaturated ; metabolism ; Mitochondrial Proteins ; physiology ; NADP ; metabolism ; Oxidation-Reduction ; Phosphotransferases (Alcohol Group Acceptor) ; physiology ; Saccharomyces cerevisiae ; growth & development ; metabolism ; Saccharomyces cerevisiae Proteins ; physiology

OBJECTIVETo investigate the value of detection of hepatitis C virus core antigen (HCV-cAg) for screening blood donor by using the internal reagent enzyme immunoassay (EIA) and anti-HCV antibody.

METHODSThe first and repeat assays were performed for detection of serum anti-HCV and HCV-cAg ELISA in 3972 donor's serum specimens from August to October of 2004. Twenty-five donors positive for anti-HCV were tested with HCV-cAg EIA kits and the results were compared with the results of HCV RNA determination with RT-PCR method.

RESULTSIn 3972 donor's serum samples, only 1 serum specimen was positive for HCV RNA identification among 10 specimens which were positive for anti-HCV in first assays, and only 1 serum specimens was positive for HCV RNA identification among 12 specimens positive for anti-HCV in repeat assays, only 2 serum specimens were positive HCV RNA identification in 3 specimens which were positive for HCV-cAg assays.

CONCLUSIONThe sensitivity of HCV-cAg ELISA is similar to HCV RT-PCR, but it is much cheaper. Therefore, HCV-cAg ELISA and anti-HCV may be used together to screen blood donor.

Blood Donors ; Enzyme-Linked Immunosorbent Assay ; Hepatitis C Antibodies ; blood ; Hepatitis C Antigens ; blood ; Humans ; RNA, Viral ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Core Proteins ; blood

Objective To-identify the factors associated with radiation-induced liver disease (RILD) and to describe the probability of RILD using the Lyman normal tissue complication(NTCP) model for primary liver carcinoma(PLC) treated with hypofractionated conformal therapy (CRT).Methods A total of 109 PLC patients treated with hypofractionated CRT were prospectively followed according to the Child-Pugh classification for liver cirrhosis,93 patients in class A and 16 in class B.The mean dose of radi- ation to the isocenter was (53.5?5.5) Gy,fractions of (4.8?0.5) Gy,with interfraction interval of 48 hours and irradiation 3 times per week.Maximal likelihood analysis yielded the best estimates of parameters of the Lyman NTCP model for all patients;Child-Pugh A and Child-Pugh B patients,respectively.Results Of all the patients,17 developed RILD (17/109),8 in Child-Pugh A(8/93 ) and 9 in Child-Pugh B(9/ 16).By multivariate analysis,only the Child-Pugh Grade of liver cirrhosis was the independent factor (P= 0.000) associated with the developing of RILD.The best estimates of the NTCP parameters for all 109 pa- tients were n=1.1,m=0.35 and TD_(50) (1)=38.5 Gy.The n,m,TD_(50) (1) estimated from patients with Child-Pugh A was 1.1,0.28,40.5 Gy,respectively,compared with 0.7,0.43,23 Gy respectively,for patients with Child-Pugh B.Conclusions Primary liver cancer patients who possess Child-Pugh B cirrho- sis would present a significantly greater susceptibility to RILD after hypofractionated CRT than patients with Child-Pugh A cirrhosis.The predominant risk factor for developing RILD is the severity of hepatic cirrhosis in the liver of PLC patients.

The selective side-chain cleavage of phytosterol to 4-androstene-3,17-dione(4-AD)and 1,4-androstadiene-3,17-dione(ADD)by Mycobacterium sp.was described.Because of the similarity in chemical structure between 4-AD and ADD,it is difficult to separate them from the fermentation broth.So far,it has been verified that the ADD can be produced by dehydrogenation of 4-AD.In this reaction,3-Ketosteriod-1-Dehydrogenase(ksdD)plays an important role.The gene knocking out method was used to solve the problem.Partial sequence of ksdD was obtained by PCR which was 631bp in length.Then,a targeting vector pUC19-MK was constructed,which was electroporate into the original strain Mycobacterium neoauru.The method of homologous recombination was used to knock out ksdD gene located in the chromosome of Mycobacterium neoauru.In this way,ksdD would lose its enzyme activity.In the result,5 transformants were screened.The experiments of steroid transformation by the transformants were carried out.The productivity of 4-AD reached 17.52% after 144h,which is 192% higher than the original strain.Meanwhile,the productivity of ADD reached 6.12%,which is 89.9% lower than the original strain.