1,3-propanediol (1,3-PD) is an important material for chemical industry, therefore, there is much interest in the production of 1,3-PD. The gene dhaT encoding 1, 3-propanediol dehydrogenase ( PDOR) of Citrobacter freundii was amplified by PCR. Sequence analysis of the similarity at the nucleotide and amino acid level between the gene encoding C. freundii PDOR and that of C. freundii ( U09771 ) were 78% and 90% , respectively. The recombinant plasmid pSE-dhaT was constructed by inserting dhaT gene into expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaT. Further more the recombinant enzyme was purifed from recombinant E. coli by Ni-nitrilotriacetate affinity chromatography followed by Sephacral S-300 gel filtration. A single obvious protein about 42kDa could be obtained by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis of recombinant enzyme. The purified enzyme was used to determined enzyme property on the substrate of propionaldehyde and 1, 3-PD. The optimal temperature and optimal pH of the purified enzyme were 37℃, 8.0 for reduction and 25℃, 10. 5 for oxidation, respectively; and the kinetic property of PDOR about Km and V max were 10. 05mmol/L, 37. 27?mol/min/mg for propionaldehyde and 1. 28mmol/L, 25. 55?mol/min/mg for 1,3-PD, respectively; The deduced dhaT gene product (388 amino acids) showed a specific reduction activity of 49. 50U/mg and oxidation activity of 79. 92U/ mg. There also have a putative iron-binding motif ( G-XX-H-X-X-A-H-X-X-G-X-X-X-X-X-P-H-G) as a fingerprint pattern in the recombinant enzyme, the motif is fully conserved among these 1, 3-propanediol dehydrogenase. It is beneficial to the researches of high producing 1, 3-propanediol by gene engineering strain.

OBJECTIVETo discuss surgical approaches of ossification of the posterior longitudinal ligament (OPLL) of cervical spine.

METHODSBetween June 2005 to July 2010, 36 patients with OPLL of cervical spine were reoperated. There were 23 male, 13 female, age from 39 to 72 years (mean 57 years). The time of the first operation to the reoperation were 4 months to 24 years, an average of 3.9 years. Among 20 patients underwent anterior corpectomy and fusion (ACD) at first operation, 14 cases combined stenosis of cervical spinal canal, 10 cases were insufficient decompression of OPLL, 5 cases injured of cervical spinal cord during the first operation, 1 case was adjacent disc herniation. Among 14 cases underwent expensive open-door laminoplasty (ELAP) at first operation, 6 cases were insufficient decompression of OPLL, 4 cases were inadequate decompressed segment, 2 cases were cervical segmental kyphosis, 2 cases were progression of OPLL combined with disc herniation. Among 2 cases underwent combined approach at first operation, 1 case was insufficient decompression of OPLL, the other was adjacent disc herniation. Their pre- and post-operative X-ray, CT and MRI were analyzed. The complications of reoperation were recorded.

RESULTThere were 30 patients followed-up, with a period of 1.5 - 4.0 years, average 1.8 years. With 36 patients, none had deterioration, 2 patients had no recovery post-reoperation, 34 patients had 31.2% Japanese Orthopedic Association score improve rate. Among 22 cases underwent ELAP at second operation, 3 cases had postoperative segmental palsy. Among 14 cases underwent ACD at second operation, 3 cases had intraoperative dural defects.

CONCLUSIONSurgical strategy for OPLL of cervical spine should consider the type of OPLL and stenosis of cervical spinal canal.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; surgery ; Reoperation ; Retrospective Studies

The present study was aimed to explore the changes of phosphorylated AMP-activated protein kinase (pAMPK) level in skeletal muscle after exposure to acute hypobaric hypoxia and exhaustive exercise. Thirty-two male Sprague-Dawley (SD) rats were randomly divided into sea level and high altitude groups. The rats in high altitude group were submitted to simulated 5 000 m of high altitude in a hypobaric chamber for 24 h, and sea level group was maintained at normal conditions. All the rats were subjected to exhaustive swimming exercise. The exhaustion time was recorded. Before and after the exercise, blood lactate and glycogen content in skeletal muscle were determined; AMPK and pAMPK levels in skeletal muscle were detected by Western blot. The results showed that the exhaustion time was significantly decreased after exposure to high altitude. At the moment of exhaustion, high altitude group had lower blood lactate concentration and higher surplus glycogen content in gastrocnemius compared with sea level group. Exhaustive exercise significantly increased the pAMPK/AMPK ratio in rat skeletal muscles from both sea level and high altitude groups. However, high altitude group showed lower pAMPK/AMPK ratio after exhaustion compared to sea level group. These results suggest that, after exposure to acute hypobaric hypoxia, the decrement in exercise capacity may not be due to running out of glycogen, accumulation of lactate or disturbance in energy status in skeletal muscle.
AMP-Activated Protein Kinases ; metabolism ; Altitude ; Animals ; Computer Simulation ; Glycogen ; metabolism ; Hypoxia ; physiopathology ; Lactic Acid ; blood ; Male ; Motor Activity ; physiology ; Muscle, Skeletal ; enzymology ; metabolism ; Phosphorylation ; Physical Exertion ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate anterior cervical plating in short-level anterior discectomy and autograft bone fusion.

METHODSEighty-one patients who underwent one- and two-level anterior cervical discectomy and fusion were randomized to 2 groups, with or without instrumentation. Among them, 55 patients were followed up. The mean follow-up time was (22 +/- 7) months. Fusion rate, disc height and cervical lordotic alignment were assessed by radiographs.

RESULTSThe improving rates were 68% in non-instrumented group and 58% in instrumented group, respectively (P > 0.05). The fusion rate was 93% in the non-instrumented group and 100% in the later one. The disc height was decreased (0.7 +/- 1.0) mm in the former group and increased (1.2 +/- 0.6) mm in the later one (P < 0.01). Although the postoperative cervical lordotic alignment was maintained better in instrumented group, the difference was not significant.

CONCLUSIONAnterior cervical plating can make good influence on the result of anterior cervical discectomy and fusion in some degree.

Adult ; Aged ; Bone Transplantation ; Cervical Vertebrae ; surgery ; Diskectomy ; instrumentation ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prospective Studies ; Spinal Diseases ; surgery ; Spinal Fusion ; methods ; Transplantation, Autologous ; Treatment Outcome

Objective:To assess the midterm clinical and radiological outcomes of internal fixation and fusion for the treatment of Hirayama disease and to evaluate the clinical significance and value of this procedure.Methods:In the study,36 patients were treated with anterior cervical internal fixation and fusion.The clinical outcomes including muscle strength and atrophy were recorded.The radiological outcomes including range of motion of cervical spine and the cross-sectional area of spinal cord at each level on MRI scan were measured before and at 3 month,1 year and 2 years follow-up time points after surgery.Results:(1) Clinical outcomes:all the patients showed no further progression of symptoms except one patient with mild progression of muscular weakness and atrophy.As the time passed by,the ratio of the patients with muscle strength and atrophy improvement increased.There were 26.5 ％ of patients in 3 months,36.0％ in 1 year and 85.7％ in 2 years who experienced muscle strength improvement.8.8％ of patients in 3 months,24.0％ in 1 year and 35.8％ in 2 years felt muscle atrophy improvement.And 12 of the 14 patients showed improved muscle strength and atrophy at the end of 2 years period follow-up.(2) Radiological outcomes:the range of motion (ROM) of C2-C7 was significantly decreased after the operation.The ROM of preoperation was 62.25° ±2.10° and that of 2 years postoperation was 13.67° ± 7.51°(P ＜ 0.01).The spinal cord was of no compression on flexion MRI.The cross-section area of spinal cord on MRI was significantly increased only at C6 level (P ＜0.05) at the end of three months follow-up.The level of increased cross-section area rose to C4-C5-C6 levels (P ＜0.01) in 1 year and to C4-C5-C6-C7 levels at the end of 2 years follow-up P ＜ 0.05).The cross-section area increased 15.60％ at C4,19.08％ at C5,21.60％ at C6 and 23.91％ at C7 with significant difference (P ＜0.05) 2 years after the operation.Conclusion:Anterior cervical internal fixation and fusion is an effective surgical treatment for Hirayama disease and may provide preferable midterm clinical and radiological outcomes.This procedure has clinical significance and value in terms of control of the progression and outcome of this disease.

BACKGROUNDS: Cervical posterior decompression surgery is used to relieve ventral compression indirectly by incorporating a backward shift of the spinal cord, and this indirect decompression is bound to be limited. This study aimed to determine the decompression limit of posterior surgery and the effect of the decompression range. METHODS: We retrospectively reviewed the data of 129 patients who underwent cervical open-door laminoplasty through 2008 to 2012 and were grouped as follows: C4-C7 (n = 11), C3-C6 (n = 61), C3-C7 (n = 32), and C2-C7 (n = 25). According to the relative location of spinal levels within a decompression range, the type of decompression at a given level was categorized as external decompression (ED; achieved at the levels located immediately external to the decompression range margin), internal decompression (ID; achieved at the levels located immediately internal to the decompression range margin), and central decompression (CD; achieved at the levels located in the center, far from the decompression range margin). The vertebral-cord distance (VCD) was used to evaluate the decompression limit. The C2-C7 angle and VCD on post-operative magnetic resonance images were analyzed and compared between groups. The relationship between VCD and decompression type was analyzed. Moreover, the relationship between the magnitude of the ventral compressive factor and the probability of post-operative residual compression at each level for different decompression ranges was studied. RESULTS: There was no significant kyphosis in cervical curvature (> -5°), and there was no significant difference among the groups (F = 2.091, P = 0.105). The VCD of a specific level depended on the decompression type of the level and followed this pattern: ED < ID < CD (P < 0.05). The decompression type of a level was sometimes affected by the decompression range. For a given magnitude of the ventral compressive factor, the probability of residual compression was lower for the group with the larger VCD at this level. CONCLUSIONS Our study suggests that the decompression range affected the decompression limit by changing the decompression type of a particular level. For a given cervical spinal level, the decompression limit significantly varied with decompression type as follows: ED < ID < CD. CD provided maximal decompression limit for a given level. A reasonable range of decompression could be determined based on the relationship between the magnitude of the ventral compressive factor and the decompression limits achieved by different decompression ranges.

Objective To isolate and identify arboviruses from mosquito pools in some regions of Liaoning province.Methods Mosquitoes were collected from Shenyang,Yingkou,Panjin,Jinzhou and Dandong cities of Liaoning province in 2006.Viruses were isolated by inoculating the specimens onto C6/ 36 and BHK-21cells.The new isolates were identified using serological and molecular biological methods.Results 5410 mosquitoes were collected from the five cities in total.Three isolates produced CPE in C6/ 36 cell and five isolates produced CPE in both C6/36 and BI-IK-21 cell.Three isolates (LN0684,LN0688 and LN0689) were identified as Banna virus and one isolate (LN0636) was identified as Getah virus.Phylogenetic analysis showed that the three Banna virus strains were clustered into the same evolution branch as the other Chinese isolates.The identity of nucleotide sequence was between 91.2% and 94.7%,compared with other Banna virus strains.The new isolated Getah virus was clustered into the same branch with the strain of South Korea (swine).The identity of nucleotide sequence was 99.2%,when comparing with the strain of South Korea and was 95% to 99% with the strains fi'om Russia,mainland of China and Taiwan region.Conehmion Eight virus isolates,including three Banna virus,one Getah virus and four unknown virus strains were isolated from mosquitoes in Liaoning province.Banna virus and Getah virus were reported for the first time in Liaoning province,while Getah virus showed the highest nucleotide homology with the South Korea strains.

BACKGROUNDChromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.

METHODSAll 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.

RESULTSThe analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.

CONCLUSIONSChinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.

Adult ; China ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology