Animals ; Biomarkers ; Hexanes ; poisoning ; toxicity ; Humans ; Neurotoxicity Syndromes ; etiology

Objective To know the expression of 47 kD protein of orientia tsutsugamushi(Ot) Karp strain and test its immunogenicity and antigenicity.Methods Forty-seven kD protein gene that code the whole mat peptide was amplified from the genomic DNA of Ot.Karp strain by PCR and constructed to a recombinant plasmid pGEX-47.The E. coli cells BL21 were transformed with pGEX-47,then the transformants were induced to express the recombinant 47 kD protein by IPTG. The antigenicity and immunogeni-(city) were identified by Dot Blot method and the experiment of immunizing mice respectively.Results 1. The PCR product of 47 kD protein gene of Ot.Karp strain was obtained and its sequence was the same as the published 47 kD protein gene.2. The E.coli expression plasmid pGEX-47 constructed was identified containing the 47 kD protein gene fragment by restrication analysis and PCR identification,and an expression band about 47 kD was detected by SDS-PAGE of the purified product from GST-fusson protein expressed by pGEX-47. 3.Dot Blot analysis and the experiment of immunizing mice testified its antigenicity and immunogenicity.Conclusions The 47 kD protein gene of Ot Karp strain is amplified that code the whole mat peptide, which can be expressed in prokaryotic cells and the expressed protein has antigenicity and immunogenicity.

Animals ; Biological Warfare ; Burns ; pathology ; Burns, Chemical ; pathology ; Chemical Warfare ; China ; Humans ; Military Medicine ; Radiation Injuries ; pathology ; Wounds, Gunshot ; pathology

OBJECTIVETo evaluate the clinical outcomes of percutaneous intervertebral foramina endoscopic lumbar discectomy for elder patients with lumbar spinal stenosis syndrome.

METHODSFrom July 2006 to July 2011, 60 elder patients with lumbar spinal stenosis syndrome were treated with surgical operation, including 32 males and 28 females with an average age of (66.7 +/- 2.5) years old ranging from 72 to 83 years. These patients were divided into the traditional surgery group and percutaneous intervertebral foramina endoscopic discectomy groups (PTED group), 30 cases in each group. The index of the preoperative and postoperative, operative incision visual analogue scale (VAS) of two groups were compared. The Oswestry disability index (ODI) of two groups at 6, 24 months of the follow-up were also evaluated on activity of daily living.

RESULTSThe average operative time, the average blood loss, the number of cases using analgesic drug, hospitalization time of PTED group were better than those of the traditional surgery group (P < 0.05). The improvement of incision VAS in PTED group was better than that in the traditional surgery group (P < 0.05). All patients were followed up for 24 months at least. The ODI at 1, 24 month after operation were better than that of preoperative in two group respectively (P < 0.05), but the improvement of PTED group was better than that of the traditional surgery group (P < 0.05).

CONCLUSIONPTED has the advantages of smaller incision, less bleeding, less postoperative stay and hospitalization time, tissue trauma and quicker recovery. It is a safe and efficacious minimally invasive surgical technique for elder patients with lumbar spinal stenosis syndrome.

Aged ; Aged, 80 and over ; Decompression, Surgical ; Diskectomy, Percutaneous ; Endoscopy ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Spinal Stenosis ; surgery ; Treatment Outcome

Objective To investigate the importance of plasmid-mediated quinolone resistance in the development of quinolone resistance in clinical isolates of gram-negative bacteria.Methods A total of 541 consecutive clinical isolates of gram-negative ba- cilli resistant or intermediate to ciprofloxacin were screened for the qnrA gene by PCR.Conjugation experiments were carried out with azide-resistant E.coli J53 as a recipient.The aac(6')-Ib-cr gene was detected.The mutations in the quinolone-resist- ance-determining region (QRDR) of the gyrA and parC genes were identified in qnrA positive strains.Results qnrA was identi- fied in 7 of the 541 strains.Among the qnrA positive strains,5 were Enterobacter cloacae.No qnrA was detected in nonfer- menters.Quinolone resistance was transferred in 4 of 7 qnrA positive strains.Transconjugants had 12-to 125-fold increases in MIC of ciprofloxacin relative to that of the recipient.Seven strains contained qnrA with a nucleotide sequence identical to that originally reported.Two transconjugants with higher ciprofloxacin MICs contained aac(6')-Ib-cr gene.Mutations occurred in the QRDR of the gyrA and parC genes in 5 PCR-positive clinical strains.Conclusions Transferable plasmid-mediated quinolone resistance associated with qnrA is highly prevalent in clinical strains of Enterobacter spp.aac(6')-Ib-cr gene and mutations in the quinolone targets may co-exist with qnrA,which may contribute to the further increase of resistance to quinolones.

OBJECTIVETo review the clinical features of the thoracolumbar fracture with ankylosing spondylitis (AS) in order to avoid delayed or missed diagnosis.

METHODSFive patients of thoracolumbar fracture with AS treated from April 2005 to June 2007 in our department were studied retrospectively, male 4 cases, female 1 case, the age from 26- to 72-years-old with an average of 44.8 years. Analysis including: case history, number of the ankylosed vertebras, characteristic of fracture, active state rheumatism.

RESULTSThe patients had the history of AS for average 22.6 years. The mean number of the ankylosed vertebras was 18.2. Of the 5 cases, 1 case encountered traffic accident, 1case was sprained, and 3 cases without trauma were diagnosed as stress fracture. Two cases were trans-vertebra fracture: the fracture line was through T6, T7, or L1 vertebral body respectively; 3 cases were through the disc space: 2 cases were through L1,2 disc space, 1 case was through L2,3. No compression fracture and neurological injury were found. The acute inflammatory index such as ESR and CRP in 4 cases didn't correlate with the degree of pain. The non-steroidal anti-inflammatory drugs (NSAIDs) hadn't significant effectiveness in relieving pain. The patients were diagnosed as 'relapse' of AS in other hospital, and had been misdiagnosed for average 1.51 months.

CONCLUSION1) the fracture is prevalent at the middle or late period of AS when extensive ankylosis has been existed at the thoracolumbar region; 2) the fracture is common at the lower thoracal spine and the upper lumbar spine, and the majority is the stress fracture; 3) the fracture line may be through the vertebral body, but more often through the disc space; 4) it is like an exacerbation of AS and therefore to be missed diagnosis; 5) when the back pain exacerbated suddenly in the middle or late period of AS, the degree of pain not correlating with acute inflammatory index, and the NSAIDs ineffective, the thoracolumbar fracture should be considered.

Aged ; Female ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fractures ; diagnosis ; drug therapy ; etiology ; Spondylitis, Ankylosing ; complications ; Thoracic Vertebrae ; injuries

OBJECTIVETo study surgical techniques for degenerative lumbar scoliosis associated with lumbar stenosis and evaluate their clinical significane.

METHODSThirty-two patients with degenerative lumbar scoliosis associated with spinal stenosis were treated by techniques of posterior lumbar interbody fusion or posterolateral fusion and pedicle screws. There were 18 male and 14 female with 56.8 years old on the average (ranging from 49 to 75 years). There were no evident change of lumberlordosis in 15 cases, and lumber lordosis were obvious loss associated with lumbar subluxation in 17 cases. The correcting, the improvement of back and leg pain, complications and followed-up results were analyzed retrospectively.

RESULTSThirty-two cases were followed-up for 6 to 39 months (the average time of 13 months). The average correction rate of scoliosis was 58.0% and the rate of pain relief was (80.2 +/- 5.8)%. There were two cases of dura sac laceration, two cases of nerve roots injury and a case of pseudoarthritis. During followed-up, correction rate and height of disc spaces were not lost. Shift of interbody cages were no displaced; all the internal fixation got well fusion and the rate of fusion for the bone graft was 96.9%.

CONCLUSIONPosterior pedicle screws combined with interbody fusion or posterolateral fusion is a safe and effective surgical treatment for degenerative lumbar scoliosis associated with lumbar stenosis.

Aged ; Bone Screws ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Scoliosis ; surgery ; Spinal Fusion ; Spinal Stenosis ; complications ; surgery ; Treatment Outcome

Objective To establish animal models with anterior transposition of the ulnar nerve and evaluate the safety of anterior transposition of the ulnar nerve at molecular level.Methods Location of the ulnar nerve of elbow in 5 rats were found similar to human being by anatomy.Twenty healthy adult SD rats,weighting about 250 g,were performed the anterior transposition of the ulnar nerve in the right forelimbs and the left forelimbs was considered as control group.The bilateral flexor carpi ulnaris muscles were weighed and the slice of cervical spinal cord(C_6-T_1)level were prepared 1 month after the operation.Nissl staining,NADPH-d histochemical staining,IB4 staining and ChAT-immunohistochemical staining were employed to observe the spinal cord(C_6-T_1)level at molecular level;electron microscope was used to observe the ultrastructure of ChAT-positive neurons.Statistical analysis was paired T test.Results The flexor carpi ulnaris muscles in the model group(92.3±9.13mg)and control group(93.2±7.29 mg)were not significantly different(P>0.05).After anterior transposition of the ulnar nerve in rats,no significant differences in cell number and morphology in the cervical spinal cord(C_6-T_1)were found between the model group and the control group(P>0.05).No changes between the 2 groups were noted in the fine structure of anterior horn motor neurons and the expression of nenrotransmitters(P>0.05).Conclusion Anterior transposition of the ulnar nerve can be safely done in the animal models(rats).

OBJECTIVETo establish serum proteome fingerprinting predictive models and search for proteins associated with colorectal cancer.

METHODSThirty-six randomly selected colorectal cancer patients and 36 cases with hernia or gall bladder diseases scheduled for elective operation were enrolled as cancer group and control group respectively. Peripheral venous blood samples were collected before the operations. Special serum protein or peptide fingerprint was investigated by using surface enhanced laser desorption/ ionization-time of flight-mass spectrometry (SELDI-TOF-MS) measurement after blood sample had been treated with weak cation exchange protein chip (CM10) for each case. The obtained data were analyzed by Biomarker Wizard software to screen serum proteome tumor markers and set up diagnosis predictive model for colorectal cancer. Blind validation of the model with 44 healthy controls and 88 colorectal cancer patients were carried out by using Biomarker Patterns Software.

RESULTSIn comparing colorectal cancer group with control group, 5 specific protein peaks (P < 0.05) were found. The predictive model had a sensitivity of 100% and a specificity of 97.2%. A sensitivity of 71.6% and a specificity of 72.7% was got with the blind validation. The specific protein peaks with a mass-to-charge ratio (m/z) of 8908 and 13,707 showed in all the results and it showed their strong relationship with colorectal cancer.

CONCLUSIONSThe predictive models built by the differences of serum proteome fingerprint could be a very useful diagnostic tool in colorectal cancer. Proteins with m/z of 8908 and 13,707 would possibly be the tumor markers of colorectal cancer.

Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Colorectal Neoplasms ; blood ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Peptide Mapping ; Proteomics ; methods ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo construct the localization system involving anti-TfR monoclonal antibody (McAb) and AFP promoters and assess its effect on human hepatoma cell lines.

METHODSThe conjugate of anti-TfR McAb and polylysine (PLL) was made by SPDP and purified by molecular screen chromatography. DNA blocking test determined that the ratio of one pEBAF/tk to six Ab-PLL was the most suitable to couple them. The pEBAF/tk recombinant plasmid bearing HSV-TK gene was coupled to Ab-PLL by noncovalent bond. The pEBAF/tk was transferred into human hepatoma cell line HepG2, SMMC7721 and pulmonary cancer cell line A549 by receptor-mediated gene delivery (Ab-PLL-DNA) and liposome procedure. The growth inhibitory rates of HepG2, SMMC7721 and A549 cells were measured by MTT assay.

RESULTSThe inhibitory rates of HepG2/tk in 100 mg/L and 1 mg/L of GCV were 60.5% and 24.3%, respectively. The inhibitory rate of GCV to SMMC7721 was 23.2% in 3 days. The pulmonary cancer cell A549, A549/tk (Ab) and A549 /tk (lipo) could not be inhibited by the addition of GCV.

CONCLUSIONThe localization system employed in this paper has high specificity, effectiveness and safety for gene therapy. It would be a promising strategy for gene therapy.

Antibodies, Monoclonal ; therapeutic use ; Carcinoma, Hepatocellular ; therapy ; Cell Line, Tumor ; Ganciclovir ; therapeutic use ; Genetic Therapy ; Humans ; Liver Neoplasms ; therapy ; Receptors, Transferrin ; immunology ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; alpha-Fetoproteins ; genetics