Objective　To establish a rapid,effective,convenient method for detecting human adenoviruses and other viral pathogens using magnetic bead separation and PCR amplification.Methods　A biotinylated oligonucleotide primers were hybridized to adenovirus DNA in stool samples.Setreptavidin coated magnetic beads were then added to isolate the DNA-oligonucleotide hybrid.The procedure allows for the recovery of viral DNA suitable for amplification by polymerase chain reaction.Results　Ten samples collected from clinical stool were detected; six of them were positive.Results indicated that this nucleic acid separation technology is very effective in concentrating and purifying adenoviruses DNA while removing PCR inhibitors in stool samples.It also effectively increase the sensitivity of PCR amplification.Conclusion　This technique can rapidly,reliably detect adenoviruses in clinical samples,and it can be used to detect other viral pathogens.

BACKGROUND: Many scholars adopt allograft interbody fusion for vertebral body resection and reconstruction, bone fusion time is better than autologous bone graft's, and its integration provides an early support and stabilizing, but the preparation of allogeneic bone graft material is easy to destroy b0ne-inducing factor in matrix, which is not conducive to bone growth. OBJECTIVE: To innovatively design and verify the ability of reconstructing rabbit cervical vertebrae with the compound of humeral cortical ring allograft (HCA) packed with red bone marrow (RBM) and autogenous cancellous bone (ACB). DESIGN, TIME AND SETTING: Randomized controlled animal experiment was performed at the laboratory of Orthopedic Department in Renmin Hospital of Wuhan University, between October 2004 and March 2006. MATERIALS: Sixty healthy adult New Zealand white rabbits, of either sex, body mass of 2.0-2.5 kg, were involved in this study. Twelve rabbits were used for HCA preparation, while the remaining 48 rabbits were randomly divided into 3 groups with sixteen rats in each group. Autologous RBM was extracted from the anterior superior lilac spine through puncture; ACB was obtained from td-cortical bone of rabbit iliac crest. Autologous RBM and ACB were compounded and filled in the self-made HCA. METHODS: Models of the fourth cervical vertebrae defect were created by surgery to simulate tumor resection in New Zealand white rabbits, which were divided into 3 groups randomly. Combined transplant group was treated with the compound of RBM+ACB+HCA; autologous bone transplant group with autogenous lilac crest; HCA transplant group with HCA. MAIN OUTCOME MEASURES: Vertebral reconstructions were evaluated by X-ray, histopathological observation and scanning electron microscope, as well as measurement of serum alkaline phosphates at different periods postoperatively. RESULTS: Eight weeks post-surgery, graft materials fused with the upper and lower cervical fusion, a large number of bone callus were observed in combined transplant group and autologous bone transplant group; HCA transplant group was present with a small amount of callus growth and poor fusion. Serum alkaline phosphatase levels were elevated in all groups, significantly higher in combined transplant group and autologous bone transplant group compared with HCA transplant group (P < 0.01 ). There were no significant differences of serum alkaline phosphatase levels between combined transplant group and autologous bone transplant group at 4 weeks or among 3 groups 8 weeks (P > 0.05). Histological analysis exhibited numerous mature bone matrix, bone trabecula and bone marrow cavity formed in combined transplant group and autologous bone transplant group. Scanning electron microscopy showed that many new bone formations in combined transplant group and autologous bone transplant group.CONCLUSION: The compound of RBM+ACB+HCA and autogenous lilac crest transplantation can efficiently reconstruct cervical vertebrae, RBM+ACB can improve the reconstruction efficiency of HCA, and could use as a matedal in cervical reconstruction.

Objective: To develop a new quantitative determination method for the biological activity of recombinant human ciliary neurotrophic factor. Methods: Dorsal root ganglions were derived from the chick embryo and dispersed into single neuron cell,The rhCNTF was added to neuron cells and incubated for 64 hours,The activity of acid phosphatase in neuron cells was determined and the biological activity of rhCNTF was analyzed quantificationally. Result: rhCNTF could promote original era dorsal root neuron cells of chick embryo surviving,the livability of neuron cells was positively related to the amount of rhCNTF added to the culture. Conclusion: A quantitative determination method for the biological activity of rhCNTF was developed by testing the activity of acid phosphatase in neuron cells. Compared with the typical ways,this method was quantificational easily,repeatable better and with much fewer disturbance factors.

Objective To evaluate the efficacy and the side effects of whole body hyperthermia(WBH) combined with chemotherapy for the treatment of advanced non-small cell lung cancer. Methods 24 patients were included in trial group(16 male and 8 female), 15 patients in Ⅲb stage and 9 patients in Ⅳ stage.6 patients had received chemotherapy before, received 1 course of WBH combined with Docetaxel; 18 cases were previously untreated, received 1 course of WBH combined with Paclitaxel and Carboplatin, then another course chemotherapy in the same time period. 26 patients were included in control group (17 male and 9 female), 16 patients in Ⅲb stage and 10 patients in Ⅳ stage. 6 patients had received chemotherapy before, were treated with Docetaxel at least 2 courses with 3 weeks interval. 20 cases, previously untreated, were treated with Paclitaxel combined with Carboplatin at least 2 courses with 3 weeks interval. Efficacy was evaluated 4 weeks after 2 courses of chemotherapy. Results The response rate was 58.3, and 30.8 in control group. The common side effects were gastrointestinal toxicity , nerve toxicity and leucopenia, but these side effects were all mild. Conclusions WBH combined with chemotherapy is an effective regimen for the treatment of advanced non-small cell lung cancer with acceptable toxicity.

Objective To explore a novel operative management for irreducible atlantoaxial dislocation. Methods Fifty-four patients, which as 32 males and 22 females aged from 7 to 63 years old with a mean of 32 years, were diagnosed with irreducible atlantoaxial dislocation, including 18 patients with os odontoideum, 22 occipitalization, 5 malunion of odontoid fracture and 9 relaxation of transverse ligament of atlas. Forty patients presented signs and symptoms of myelopathy or spinal cord injury. All of the patients underwent the operation of open reduction and release by transoral approach. The longus collies muscles, longus capitis muscles, anterior longitudinal ligament, atlantoaxial articular capsules, the apical odontoid ligament and alar odontoid ligaments may be included, were transected, followed by arthrodesis via posterior approach in one stage. Different methods of posterior arthrodesis were conducted as followed: C1,2 transarticular screw fixation in 5 cases, occipitocervical fixation using pedicle screws of axis and occipitocervical plate in 37 cases, and C1,2 joint fixation with plates and screws in the lateral masses of the atlas and axis in 12 cases. Results A complete reduction was achieved in 41 cases, and the other 13 patients obtained partial reduction. Forty-eight patients were followed up from 4 to 40 months, 15.7 months at the average. All of them achieved solid arthrodesis. According to Odom's scoring system, among the 38 patients with preoperative upper cervical myelopathy, 15 patients were assessed as excellent, 14 good, 8 fair and 1 poor. During the operation, cerebral spinal fluid leakage occurred in 1 case, and pedicle screwing failed in 1 case. After the operation, respiratory failure was found in 1 case, nasal sound in 3 cases, dysphagia in 1 case. One patient became quadriplegia due to myelitis caused by septicemia at two weeks after the operation. The internal fixation loosened in 1 case at 2 months postoperatively. Conclusion Transorally surgical release for reduction and posterior arthrodeses could achieve satisfactory outcomes in patients with irreducible atlantoaxial dislocation.

Objective To compare the consistency and accuracy of determination of peripheral blood lymphocyte(PBL)interleukin-2(IL-2)with ELISA and evaluation of the expression of PBL IL-2 mRNA with real time PCR(RT-PCR).Methods In 20 kidney recipients,the PBL IL-2 level was determined with ELISA and the mRNA expression in PBL IL-2 was evaluated by real time fluorescence quantitative PCR(RT-FQ-PCR)before operation and C0/C2 time on the 7th day after operation.The same detection and evaluation were also done in 15 chronic allograft nephropathy(CAN)patients and 5 non CAN patients.Results IL-2 and IL-2 mRNA expression showed positive correlation before operation and 1 year later after operation,R2=0.58 and 0.4 respectively,both P0.05.On the 7th day after operation,IL-2 and the inhibition rate of IL-2 mRNA expression were(16.31?9.59)% and(69.92?7.10)%,respectively.A positive correlation existed between C2 IL-2 concentration and the inhibition rate of IL-2 mRNA expression,R2=0.56,P0.05.Also,IL-2 inhibition and PBL IL-2 mRNA inhibition showed no linear correlation.Conclusions The technique of determining PBL IL-2 by ELISA can be used for evaluating immunity status and immunosuppressive tolerance in patients with renal transplantation before the operation and at stable stage after operation.However,it should not be used for assessing perioperative immunity status and evaluating effect of cyclosporine.

Objective To investigate the level and value of serum uric acid (UA) in patients with multiple system atrophy (MSA).Methods One hundred and one MSA patients were selected as the MSA group and 101 age and sex-matched healthy controls as control group.The UA concentrations and other biochemical indexes were measured by an enzymatic method and carefully analyzed.By comparing the UA concentrations for different genders, subtypes, and different levels of certainty of the MSA group with their counterparts in the control group, we obtained the differences between these two groups and also discussed possible reasons and factors.Results The serum UA levels of MSA patients ((283.74 ± 87.76) μmol/L) were significantly lower than the controls ((317.86 ± 76.95) μmol/L;t =-2.94, P ＜ 0.05).In addition, when the male and female groups were investigated separately, the decrease of serum UA was more significant in the male group compared to the female group (t =-3.88, P ＜ 0.01).Furthermore, by comparing different subtypes of the MSA patients with the control group, we found that the serum UA levels in both P-type or C-type MSA patients were significantly lower than in controls (t =2.92, 2.02;all P ＜ 0.05).By comparing different levels of MSA with the control group, we obtained that UA levels in both probable MSA patients and possible MSA patients were significantly lower than in controls (t =3.13, 2.09;all P ＜ 0.05).By adjusting for the confounding factors, such as age, alanine aminotransferase, aspartate aminotransferase, urea nitrogen, creatinine, high density lipoprotein-cholesterol and low density lipoprotein-cholesterol, the multiple Logistic regression analysis showed that the numbers of MSA patients in the lowest level UA group (UA ＜ 263.0 μmol/L) were higher than the high level UA group (UA ＞ 370.5 μmol/L), though the difference did not reach statistical significance (P ＞ 0.05).Nevertheless, the numbers of male MSA patients in the lowest level and the lower level UA groups (UA =263.0-309.0 μmol/L) were 11.5 and 10.8 times that in the high level UA group (P =0.001, P =0.003).No obviously linear relation between the level of UA and disease progression was found.Conclusions The level of serum UA is significantly decreased in MSA patients, especially in the male patients.The UA, as an antioxidant, may be beneficial to the neurodegenerative diseases.

Objective:To construct pIRES-I-Ad?? bicistronic eukaryotic expression vector. Methods: Total RNA was acquired by TRIZOL method, the genes of I-Ad ?? chains were amplified through RT-PCR, respectively. The target genes were connected to pGEM-T vector and sequenced. Then the target genes were subcloned into pIRES bicistronic eukaiyotic expression vector and NIH3T3 cell line was transfected. Transfectant was screened by G418 antibiotics. Total RNA of transfectant was obtained by TRIZOL method, mRNA of foreign gene was examined by RT-PCR. Flow cytometry( FCM) was used to detennine foreign gene expression in protein level and surface expression in NIH3T3 cell line. Results: Bicistronic eukaryotic expression vector pIRES-I-AdaB was established. Foreign gene in mRNA level in transfectants was examined. Verified Ⅰ-Ad ?? chain wa3 expressed in high level expressed on transfected NIH3T3 cell line surface by FCM. Conclusion:Bicistronic eukaryotic expression vector pIRES-I-Ad ?? was constructed successfully. It is useful for studying antigen presentation and interaction between epitopes and MHC- Ⅱ molecule of BALB/c mouse.

Adolescent ; Diagnostic Errors ; Foreign Bodies ; diagnosis ; Humans ; Male ; Trachea

Female ; Genetic Predisposition to Disease ; HSP70 Heat-Shock Proteins ; genetics ; Humans ; Lung Neoplasms ; genetics ; Male ; Polymorphism, Genetic