Objective To study the hemoprotective effects of a rotary magnetic field(RMF)with radiation- injured mice.Methods C57 BL6/J mice were randomly divided into a control group and a magnetic treatment group.The mice received total body irradiation with 7.0 Gy and 6.5 Gy ~(137)Cs?rays.The treatment group was trea- ted with a RMF for one hour at a time,twice a day.The intensity of the RMF was 0.6T.The survival rate was ob- served for 30 days.On day 7,10,14,21,28 after irradiation,the subjects' peripheral blood cells were counted.On day 12 and 16,the number of bone marrow mononuelear cells(BMNCs)was measured and their ability to form granu- locyte-macruphage colony-forming unit(CFU-GM)was assessed.The pathological sectioning of the femur was per- formed and the expression level of bone morphogenetic proteins(BMPs)in the bone marrow were evaluated.Re- sults The RMF treatment increased the survival rate and duration among the irradiated mice and the number of blood cells in their peripheral blood.Also,RMF treatment could increase the number of BMNCs and improve their ability to form CFU-GM on days 12-16.Furthermore,RMF could improve angiogenesis and the expression level of BMPs. Conclusion The RMF treatment had an obvious protective effect against the effects of irradiation,and it accelerated the recovery of hematopoiesis and the hematopoietic microenviroment in mouse bone marrow.

ObjectiveTo study the hemoprotective effects of rotating and stationary magnetic field(RSMF) in mice treated with 5-Fluorouracil(5-FU).MethodsThe BalB/C mice were randomly divided into control group and RSMF-treated group.The mice were injected with 5-FU,the dose were 150,180,210,250 mg/kg respectively. RSMF-treated group was exposed to RSMF for 1 h a time,twice a day during 30 consecutive days,and the magnetic intensity was 0.6 T.The survival rate and survival days during the 30 days were observed.7,10,14,21,28 days after injection,the peripheral blood cells were counted.On day 8,10 and 14,the number of bone marrow mononuclear cells(BMNC) and the forming ability of colony-forming unit-granulocyte/macrophage(CFU-GM) were measured.The pathological section of femur and the expression level of bone morphogenic proteins(BMPs) in bone marrow were evaluated.ResultsRSMF could increase the survival rate and survival days of mice treated with 5-FU,and induce an increase in hemoglobin concentration,white blood cell count(WBC),red blood cell count(RBC) and platelet number.Also,RSMF could increase the number of BMNC and improve the forming ability of CFU-GM on days 8～14.Furthermore,RSMF could improve the bone marrow angiogenesis and the expression level of BMPs.ConclusionRSMF have an obvious protective effect against chemotherapeutic injury,and it can accelerate the recovery of hematopoiesis and hematopoietic microenvironment in mouse bone marrow.

ObjectiveTo evaluate the effectiveness at anterior lumbar interbody fusion (ALIF) with a single cage alone in treatment of lumbar degenerative disc disease(DDD).MethodsThirty-four patients aged from 26 to 67 years (mean,52) were enrolled in this study,including 10 males and 24 females.Preoperative diagnosis included:degenerative spondylolithesis in 11 cases,discogenic low back pain in 21,and revision for failed posterior surgery in 2.Objective level located in L4.5 in 12 cases,L5S1 in 20,and double discs(L4.5,L5S1) in 2.All the patients received extraperitoneal approach ALIF.Surgical time,blood loss,perioperative complications and postoperative ambulation were noted.Postoperative function status was evaluated by 100 mm Visual Analog Scale (VAS) and Oswestry Disability Index (ODI50).AP,lateral and dynamic X-rays were routinely taken during follow up,according to which disc height,migration of implant and bony fusion were measured.ResultsAll the surgeries were performed successfully with a mean surgical time of 90 min and blood loss of 200 ml.An injury to the bifurcation of vena cave was encountered in an early case,after meticulous repairing of the vessel,the surgery was finished successfully.Peritoneum rupture was noticed in one case.One case of postoperative cage migration was revised and reinforced with additional fixation system.All the patients were followed up for an average of 36 months(range,12-60).All the patients were satisfied with the pain relief and returned to normal social life and working.VAS decreased from preoperative 70 to postoperative 5,and ODI decreased from 41％ to 6％.All but one migration case reached sound fusion with a fusion rate of 97％.ConclusionOwing to multiple advantages,complete removal of degenerative disc,compressive bone graft,sufficient graft area,early ambulation,less complication,extrapefitoneal approach for ALIF comes up with satisfying clinical outcomes.This mini_invasive technique provides a valuable alternative to treatment of lumbar DDD when appropriate indication is selected.

Objective To discuss the value of thoracolumbar injury classification and severity score (TLICS) and load-sharing scores in guiding selection of the indications of combined anteroposterior surgery for thoracolumbar fractures. Methods A total of 216 patients with thoracolumbar fractures treated surgically from January 2006 to January 2008 were involved in the study. Combined anterior and posterior surgery was carried out in 48 patients including 32 males and 16 females (at average age of 39 years, range 18-55 years). Segments involved T11 in two patients, T12 in eight, L1 in 20 and L2 in 18. According to the classification of Magerl, there were 20 patients with type B1 fractures, 15 with type B2, four with type C1 and nine with type C2. Neurologic status based on ASIA classification was at grade A in five patients, grade B in 16, grade C in 16, grade D in nine and grade E in two. All patients were operated in lateral position at one stage within 14 days after injury, with posterior laminectomy and pedicle fixation, anterior corpectomy, reduction and strut graft. Forty-five patients were followed up for 14-38 months. Plain X-ray radiographs and reconstruction CT were taken to observe the bone healing. Local kyphosis and vertebral canal were also measured. All the patients were evaluated with TLICS and load-sharing scores. Results Lumbar physical lordosis was reconstructed, with no evident correction loss,pseudoarthrosis or implant failure noted during follow-up. The patients treated with combined surgery had TLICS score ≥5 points, load-sharing scores ≥7 points and were combined with posterior ligamentous complex injury, so the fracture could not be sufficiently decompressed and reconstructed with single anterior or posterior surgery. Of all, 45 patients were followed up for 14-38 months (mean 25 months), which showed that all the combined surgeries were performed successfully, with bone fusion and neurological status improved for at least one ASIA grade. No complications like implant breakage, loosening, titanium mesh displacement or subsidence were observed. Conclusions LICS is basically helpful for guiding selection of combined surgery, but does not well evaluate the canal compromise, to which the load-sharing scores can supplement. These two evaluation systems should be applied together. The anteroposterior surgery can be recommended when the patients are with TLICS≥5 points, load-sharing score ≥7 points and combined with ligament complex injury.

The interaction among collagen, von Willebrand factor (vWF) and glycoprotein Ib axis is the first step in hemostasis and thrombosis, especially under high shear condition. To develop a new remedy of anti-thrombosis, mRNA from endothelial cells was extracted, and reverse transcription PCR was adopted to amplify DNA of interest. After sequencing, recombinant expression vector was constructed. The amplified DNA fragment of vWF domain A3 was inserted into expression vector with 6 x his taq, pET20b(+), the recombinant was transformed into E coli (strain DE3) and induced by IPTG. Recombinant vWF-A3 was designated as a recombinant fragment comprising residues 918 - 1114 of mature vWF subunit. It was purified through Ni-NTA resin column and refolded in Tris buffer containing GSH and GSSG. The results showed that rvWF-A3 was expressed successfully in E coli (strain DE3), accounting for 46% of total bacterial protein with its purity of over 95%. It was identified that rvWF-A3 is capable to bind collagen and inhibit the wild vWF binding to collagen by competition. It is concluded that rvWF-A3 fragment might be an effective antithrombotic agent for preventing arterial thrombosis.
Cloning, Molecular ; Collagen ; metabolism ; Escherichia coli ; genetics ; Humans ; Protein Structure, Tertiary ; Recombinant Proteins ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; von Willebrand Factor ; chemistry ; genetics ; metabolism

Objective To evaluate the mid-term clinical and radiographic results of artificial disc replacement (ADR) for discogenic low back pain.Methods From July 2004 to July 2007,21 patients with discogenic low back pain,aged from 26 to 67 years,underwent lumbar ADR with the Charité Ⅲ artificial disc at 22 levels and Activ L prosthesis at 2 levels:L4-5 5 cases,L5S1 13 cases,L4-5 and L5S1 3 cases.The diagnosis was proved by discography in all the patients.The clinical and radiographic results were evaluated and compared between pre-,post-operation and finial follow-up.Results All patients were followed up for 4to 7 years (average,5.1 years).There were statistical differences between pre-operation and final follow-up in Oswestry disability index and Visual Analogue Scale for back pain and leg pain.All the prostheses were mobile without dislocation,breakage,subsidence or spontaneous fusion,only a slight scoliosis was noted in one patient.At the ADR levels,the anterior and posterior intervertebral height increased averagely 6.3 mm and 1.9 mm respectively,the lordosis increased 2.9°,and the mean range of motion (ROM) was 4.6°.At the adjacent level proximal to ADR,the intervertebral height and lordosis decreased slightly,and the ROM decreased 2.5°.The total lumbar lordosis increased 7.8°,and the total lumbar ROM increased 2.4°.According to MRI and CT scans,the degeneration of proximal adjacent disc and facet were not evident,however the degeneration of facets at the ADR levels was evident.Conclusion The 5 years results of ADR for discogenic low back pain were satisfactory,with preserved motion at the ADR level,and the degeneration of adjacent level was not evident.However,there was obvious degeneration in facet joints of the ADR level.

Paf1 complex was identified in yeast and characterized to function in transcription and its related events. We identified the Drosophila homological components of paf1, CDC73 and RTF1 of paf1 complex. The genes encoding Drosophila paf1, CDC73 and RTF1 were cloned and expressed. With the purified recombinant proteins of truncated components of paf1 complex, antibodies against the Drosophila paf1, CDC73 and RTF1 were generated. These antibodies have been shown to be able to detect the endogenous paf1 subunits as well as their human counterparts in the HeLa extract. On Drosophila polytene chromosomes, these antibodies have been demonstrated to locate the paf1 complex at actively transcribing sites, which co-localized with phosphorylated RNA polymerase II, indicating that paf1 complex in Drosophila is involved in transcription or the events coupling with transcription.
Animals ; Antibodies ; chemistry ; Drosophila Proteins ; immunology ; Drosophila melanogaster

This study was aimed to further investigate the function of platelet collagen receptor-glycoprotein VI and to screen its specific inhibitor. The extracellular domain of platelet glycoprotein VI (GPVI) in E. coli was expressed by recombinant technology, the extracellular domain cDNA of GPVI was amplified from pBluescript KS(-)-GPVI plasmid by PCR. Proved by sequencing, the expression vector pET-20b(+)-GPVI was constructed, which was then transformed into E. coli (BL21(DE3)pLysS) and induced by IPTG. The recombinant GPVI was purified on Ni-NTA resin column and renatured in PBS containing GSH and GSSG. The anti-penta His McAb and anti-GPVI polyclonal antibody were used to identify the recombinant GPVI in Western blotting. Collagen binding test was conducted to investigate the biological activity of recombinant GPVI. The results showed that the recombinant GPVI was expressed in E. coli and successfully purified, which was confirmed to be similar to the native GPVI in Western blotting. The recombinant GPVI can bind the type I collagen in dose-dependent manner. In conclusion, the recombinant GPVI can be achieved in E. coli and restore its native characteristics after renaturation.
Blood Platelets ; metabolism ; Blotting, Western ; Escherichia coli ; genetics ; Humans ; Integrin alpha2beta1 ; Platelet Membrane Glycoproteins ; biosynthesis ; genetics ; Protein Binding ; Receptors, Collagen ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; isolation & purification

Senile dementia is a disease that gradually develops with age. At present, the etiology, pathogenesis and treatment of senile dementia cannot be completely determined clinically. Therefore, it is extremely necessary to prevent and treat senile dementia from prevention and disease management. For the non-ill people, this article analyzed the susceptible groups of senile dementia from the perspective of TCM constitution, combined with the current prevention plan of Alzheimer's disease, from the aspects of examination, nutritional supplementation, adjustment of work and rest, exercise, etc., early physical fitness prevention targeting populations at different stages was conducted; the early onset of senile dementia is not obvious, and the early prevention of senile dementia can be carried out from various aspects in combination with constitution identification and inspection of senile dementia; for the exact sick population, disease management guidance can be added on the basis of the above aspects to provide definite and feasible recommendations for disease prevention and management of the elderly.

BACKGROUNDDihydropyrimidine dehydrogenase (DPD), a key enzyme involved in the catabolism of 5-fluorouracil (5-FU), is the attractive candidate for pharmacogenetic research on efficacies and toxicities of 5-FU. The aim of this study is to explore the association between polymorphisms of dihydropyrimidine dehydrogenase gene (DPYD) and clinical outcomes of gastric cancer patients treated with fluorouracil-based adjuvant chemotherapy in the Chinese population.

METHODSThree hundred and sixty-two patients with gastric cancer in the Chinese population were treated with fluorouracil-based adjuvant chemotherapy. The single nucleotide polymorphic genotypes of DPYD were determined by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) using DNA samples isolated from peripheral blood collected before treatment.

RESULTSThe average response rate for chemotherapy was 46.7%. A significantly different distribution of the rs1801159 (c2=8.76, P=0.012) genotypes was observed. Homozygous genotype rs1801159A/A was over-represented in responsive patients. Conversely, carriers of the rs1801159A/G genotype were prevalent in non-responsive patients. In the haplotype association analysis, there was significant difference in global haplotype distribution between the groups (c2=3.96, P=0.0465).

CONCLUSIONSThese results suggest that polymorphisms of rs1801159 in DPYD may be used as valuable predictors of the response to fluorouracil-based chemotherapy for gastric cancer patients in the Chinese population. Well-designed, comprehensive, and prospective studies on determining these polymorphisms of DPYD as predictive markers for gastric cancer in response to fluorouracil-based therapies are warranted.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; Chemotherapy, Adjuvant ; methods ; Dihydrouracil Dehydrogenase (NADP) ; genetics ; Female ; Fluorouracil ; therapeutic use ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Stomach Neoplasms ; drug therapy ; genetics ; Treatment Outcome ; Young Adult