ObjectiveTo study the features of lower limb vein thrombus patients after spinal cord injury and the effects of thrombolysis therapy.MethodsWe practiced ultrasonography on 207 patients with lower limb swelling after spinal cord injury. ResultsAmong the 207 patients, 66 had thrombus, the positive rate was 31%: acute thrombus 58 cases, subacute thrombus 1 cases and chronic thrombus 7 cases. Conclusions Ultrasonography provides an important basis for the qualitative diagnoses of lower limb vein thrombus. It has certain clinical guiding effects in choosing methods of treatment.

Objective To assess the efficiency of total laminectomy and transpedicular screw placement techniques and a thoroughly decompression with a three column fixation device on the early cervical spine cord injury.Methods 27 patients were operated successfully with total cervical laminectomy through transpedicular screw techniques.Precise measurements of all cervical spines were made by postoperative CT scan of pedicle dimensions,angulation,and offset relative to the lateral mass boundaries.And meanwhile,cortical integrity and neurovascular injury were then assessed by obtaining postoperative computed tomography scans of each patient.Results Using manual techniques,220 pedicles were instrumented and all the cases were followed up between three to twenty-four months.The postoperative gradations were obviously better than the preoperative evaluation.Regardless of the technique used,the vertebral artery was the structure most likely to be injured.Conclusion Insufficient correlation between different surgeons' assessments of surface landmarks attests to the inadequacy of screw insertion techniques in the cervical spine based on such specific topographic.At the early stage of cervical spinal cord injury,using total laminonectomy decompression through cervical screws techniques to reconstruct spinal alignment could improve the prognosis of patients.

An oxygen-tolerant denitrifying strain designated as H1 was screened by the procedures of shallow shaking and continuous aeration cultures.With the aid of an nnrS-gfp fusion responsive to nitric oxide (NO)and acetylene inhibition-GC procedure,it was shown that strain H1 was able to produce NO and N_2O but not N_2 under denitrifying conditions.Denitrifying processes were thus determined as NO_3~-→NO_2~-→NO→N_2O,with N_2O as the end product.Strain H1 could denitrify under shallow shaking conditions as well as in the initial atmospheric oxygen concentration ranging from 0～21%.Denitrification processed normally under continuous aeration at the rate of 2 L air per min in a 150 mL medium,but stopped under high aeration rate as 5 L air per min.16S rRNA gene sequence revealed that strain H1 shared 98% similarity to its closet relative Ralstonia taiwanensis,the genus where denitrifying bacteria are frequently found.

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.
Animals ; Antibodies, Bispecific ; chemistry ; Antibodies, Monoclonal ; chemistry ; Arthritis, Experimental ; Escherichia coli ; Fibronectins ; chemistry ; Half-Life ; Humans ; Mice ; Single-Chain Antibodies ; chemistry ; Tumor Necrosis Factor-alpha ; chemistry

Objective To investigate the prevalence of plasmid-mediated quinolone resistance ( PMQR ) determinants [ qnr,aac ( 6' ) -Ib-cr and qepA ]and mutations in quinolone resistance-determining regions (QRDRs) of gyrA and parC and their association with fluoroquinolone susceptibility in clinical isolates of Serratia marcescens in Anhui.Methods The minimum inhibition concentration ( MIC ) of 104 strains of S.rnarcescens collected from various clinical specimens from 34 hospitals during 2005 to 2010 were determined by agar dilution method.The qnr,aac (6')-Ib,qepA,gyrA and parC genes were screened by polymerase chain reaction (PCR) in 31 strains resistant to ciprofloxacin,and positive results were subsequently confirmed by sequencing.The conjugation experiments were performed for qnr and aac(6')-Ib-cr positive strains.The MIC of S.marcescens isolates,recipient strains and conjugants were tested by agar dilution method for quinolones and other antimicrobial agents.Results Six strains of the 31 S.marcescens isolates harboured qnr and/or aac(6')-Ib-cr genes.Among those 6 strains,2 strains harboured a qnrB6 gene,1 harboured a qnrS2 gene,and 4 harboured aac( 6' ) -Ib-cr,whereas no qnrA-,qnrC- or qnrD-positive isolate was detected.None of the 31 isolates carried the qepA gene.Mutations in the QRDR of gyrA and parC genes were detected in 9 and 7 isolates,respectively.The conjugation experiments were successfully carried out in 5 isolates of 6 PMQR determinants-postive strains.The MIC of conjugants for quinolones were increased evidently compared to recipient strains.Conclusions Chromosome and plasmid-mediated resistance determinants play an important role in quinolone resistance in clinical isolates of S.marcescens.And more important is that the PMQR determinants can be horizontal transmitted.It is necessary to continuously survey and watch for the spread of PMQR in S.marcescens in public health control program.

Objective To evaluate the results of rotary self-locking intramedullary nail (RSIN) for treating long tubular bone fracture of extremities, and discuss the current problems. Methods One hundred and twenty-two patients with long tubular bone fracture of extremities,including 59 femoral fractures,57 tibial fractures and 6 humeral fractures,who had been treated by RSIN were retrospectively investigated. Results All of the patients achieved clin-ical healing,with an average time of 24 weeks. Nobody was found to appear maluniun,infection and the break of inter-nal fixture. Internal fixations were removed after the fracture healing,with an average time of 14 months. It was diffi-cult to remove the internal fixations in 5 cases, and one case refractured after removing the internal fixations. Conclu-sion RSIN has the advantages such as easy operation,less trauma,no pendulum effect,early motion after operation and so on, except the characters of general interlocking intramedullary nail such as anti-rotation, anti-crispition and anti-displacement.

OBJECTIVEScreening and characterizing high affinity completely humanized single-chain antibody (ScFv) against PreS1 of hepatitis B virus.

METHODSA combinatorial library of phage-displayed human ScFv, genes of which were derived from peripheral blood lymphocytes immunized by PreS1 of Hepatitis B Virus in vitro, was constructed. The library contained 7 10(8) clones.

RESULTSAfter 3 rounds panning, a high affinity (K=10(7) to 10(8) mol/L) ScFv specific to PreS1 was obtained. The V(H) belonged to human V(H4) family, and V(1) to V(4) by sequence analysis.

CONCLUSIONThis study suggests that the method of antigen stimulation in vitro is an expeditious way for the source of human immune antibody. And the ScFv may provide a more satisfactory therapy.

Antibodies, Monoclonal ; biosynthesis ; Antibody Affinity ; Hepatitis B Antibodies ; biosynthesis ; Hepatitis B Surface Antigens ; immunology ; Humans ; Peptide Library ; Protein Precursors ; immunology

Objective To study the exocrine pancreatic function in critically ill children with septic shock,sepsis and hyperlactacidemia.Methods A total of 64 critical pediatric patients were admitted from Jan 2009 to Oct 2012,and clinical and laboratory findings including pancreatic function,and histopathological features and score after autopsy were reviewed.Results (1) Compared with non-septic shock children,the pancreatic pathology score and serum lipase in septic shock group were significantly higher and serum calcium was significantly reduced (P ＜0.05) ; (2) The pancreatic histopathology score was significantly increased in patients with elevated plasma lactate ≥2 times (P ＜0.05),but there were no significant differences in serum calcium and blood amylase and lipase between patients with elevated plasma lactate level and patients with normal plasma lactate level; (3) The concentrations of serum amylase,lipase and urinary amylase were significantly increased in patients with pancreatic histopathology score ＞4 points compared with score ≤4 points patients,but there were no significant differences in above three biomarkers between patients with score ≤3 points and patients with score ＞3 points.Conclusions The pancreas is vulnerable to damage easily occurred in septic shock children especially complicated with hyperlactacidemia.The pancreatic histopathology score ＞ 4 points can be as a sensitive and reliable indicator of pancreas damage.

Objective To report two childhood acute myeloid leukemia (AML) patients with t(8;20)(q22;q13) and t(1;8;21)(q32;q22;q22) respectively,as variant t(8；21).Methods Chromosome preparation of bone marrow cells were made using short-term culture and karyotypic analysis was carried out using R and G-banding techniques.Interphase-fluorescence in situ hybridization (I-FISH) and metaphase-FISH (M-FISH) were performed using dual color,dual fusion AML1-ETO probe to detect the AML1-ETO fusion gene.Multiplex RT-PCR was used to demonstrate the expression of AML1-ETO fusion transcript.Results The karyotype of bone marrow cells for these two childhood AML patients were 45,X,-Y,t(8;20)(q22;q13)[12]/46,XY[3](case 1) and 46,XX,t(1;8;21)(q32；q22;q22)[18]/46,XX[2](case 2),respectively.I-FISH and M-FISH confirmed that they all had the AML1-ETO fusion gene and variant t(8;21).The AML1-ETO fusion transcript in both patients was detected by RT-PCR.Conclusion t (8;20)(q22;q13) and t (1;8;21)(q32;q22;q22) are variant t (8;21) in nature.It is important to combine the conventional karyotypic analysis with D-FISH and multiplex RT-PCR to determine the nature and prognosis of AML patients with variant t(8;21).

ObjectiveTo observe the effect of Huanglian Wendan decoction and Danshen decoction combined with Wester medicineon stable coronary heart disease.Methods60 patients with angina or myocardial infarction were randomly divided into the control group and treatment group with 30 cases in each group. The patients of the control group were treated with routine medicine and those of the treatment group were added with traditional Chinese medicine (TCM) Huanglian Wendan decoction combined with Danshen decoction. The changes of angina, symptoms and signs of TCM, electrocardiogram (ECG), low density lipoprotein cholesterol (LDL-C) and high sensitive C-reactive protein (HS-CRP) of all patients were observed before and after treatment.ResultsAfter treatment, the frequency and duration of angina, dose of notroglycerine of patients in two groups were significantly different from those before treatment (P<0.01 ), but there was no significant difference between two groups (P>0.05). The symptoms such as chest discomfort and palpitation of patients of two groups improved after treatment (P<0.05～0.01), but the treatment group improved more obviously than the control group (P<0.05). The total effective rate showed by ECG were 53.3% in the control group and 73.3 % in the treatment group, there was no significant difference between two groups. HS-CRP and LDL-C of all patients decreased after treatment (P<0.01), but the treatment group was lower than the control group (P<0.05).Conclusion Huanglian Wendan decoction combined with Danshen decoction can relief clinical symptoms of stable coronary heart disease, which may be related with lipid decrease and inflammation reduction.