Objective:To choose the best vector for the expression of CS3 fimbriae. Methods: The CS3 operon was cloned into different plasmid vectors such as pUC19 and pTrc99A. The expression of CS3 was monitored by whole-cell ELISA and SDS-PAGE analysis. The assembly of CS3 fimbriae was detected with electron microscopy. Results:The expression level of CS3 fimbriae using plasmid pUC19 as carrying vector was the highest, and the insertion orientation of CS3 gene into the plasmid has a little effect on its expression level. The expression of CS3 fimbriae was confirmed by SDS-PAGE analysis and electron microscopy.Conclusions:The promotor of CS3 itself played the key role in the expression of CS3 fimbriae and the copy number of plasmid was the main factor to affect the expression level.

Objective To report the clinical effect of free transplanting for soft tissue defects pedieled with high site cutaneous branches of the transverse branch of lateral cirumflex femoral artery.Methods Cu- taneous branches of the descending branch of lateral circumflex femoral artery were found small or abscent in 7 patients.The anterolateral femoral skin flap was pedicled with high site cutaneous branches of the transverse branch to repair the soft tissue defects of the arm,hand,leg and foot,rather than with the descending ones. The size of the flap ranged from 15 cm?6 cm to 28 cm?13 cm,with part muscle valve,iliotibal tract and lat- eral femoral cutaneous nerve.The fractures were performed with internal or external fixation.Results All of the anterolateral femoral skin flap survived well postoperatively in the 7 cases and had good appearance and sensation at one stage.The function of the repaired extremities recovered well.Conclusion The anterolat- eral femoral skin flap pedicled with high site cutaneous branches of the transuvrse branch of lateral circumflex femoral artery has many advantages of good blood supply and large size.The flap was secluding,and can be taken with some muscle and lateral femoral cutaneous nerve.When cutaneous branches of the descending branch of lateral circumflex femoral artery is small or abscent,the anterolateral femoral skin flap with high site cutaneous branches of the transverse branch of lateral circumflex femoral artery is an optimal alternative.

The intracellular calcium signaling, which is modulated by the microenvironment of cells, is closely related to the cell's self renewal, differentiation, proliferation, and its apoptosis. The study on the quantitative modulation of the intracellular calcium signals could not only help to understand the dynamic behavior of such kind of signaling, but also play a significant role in the control of cell fate, simulation of cell behavior and bionics of cellular biological systems. This paper briefly reviewed the progress on the quantitative modulation of intracellular calcium signals induced by shear flow, including (1) experimental phenomena and the associated mechanisms of shear flow activated intracellular calcium response; (2) mathematical modeling and simulation of the intracellular calcium response induced by shear flow; (3) feedback control of the intracellular calcium signals by shear flow.

OBJECTIVETo evaluate the value of real time quantitative RT-PCR (Q-PCR) for monitoring AML1-ETO mRNA levels in AML1-ETO(+) acute myeloid leukemia (AML) patients following allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSQuantification of AML1-ETO(+) mRNA was performed serially on bone marrow samples from 17 patients with AML1-ETO(+) AML after HSCT. Q-PCR used the TagMan probe system. The AML1-ETO mRNA level was normalized by control gene abl. Cytogenetic response was evaluated by fluorescent in situ hybridization (FISH).

RESULTSThe reproducible sensitivity of Q-PCR was 5 copies. Out of 16 patients who achieved sustained complete cytogenetic response (CCyR), one each died of graft-versus-host disease and infection. The median AML1-ETO mRNA levels in the rest of 14 CCyR patients were 0 (0 - 0.740), 0.026 (0 - 2.900), 0.039 (0 - 3.300) at +1, +2, +3 month post allo-HSCT, respectively and in 5 CCyR patients beyond 1 year following allo-HSCT (median follow-up 685 days) was 0.078 (0.003 - 0.120). The AML1-ETO mRNA levels in one relapsed patient were 0, 9.8 and 5.6 at +1, +2 and +3 month post allo-HSCT, respectively and hematological relapse occurred at +110 day, when the AML1-ETO mRNA levels increased dramatically from 5.600 to 390.000.

CONCLUSIONSQ-PCR is a sensitive technique in monitoring AML1-ETO(+) AML patients post allo-HSCT. Persistence of a low level within one year after allo-HSCT does not mean at risk of relapse. It is necessary to dynamic monitoring AML1-ETO mRNA after remission in t(8;21) AML patients.

Adolescent ; Adult ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; surgery ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; metabolism ; RNA, Messenger ; genetics ; RUNX1 Translocation Partner 1 Protein ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Young Adult

OBJECTIVETo explore dynamic status of bcr-abl transcript levels in chronic myeloid leukemia (CML) patients after allogeneic HSCT and to define low relapse patients who may not require further therapeutic intervention.

METHODSOne hundred and forty-nine CML cases received allo-HSCT at our Institute between Nov. 2004 and Nov. 2006. Bcr-abl transcripts levels were monitored from bone marrow samples by Q-RT-PCR in all cases pre-HSCT and at 1, 2, 3, 4, 5, 6 months post HSCT. The median value of bcr-abl at these time points were calculated and compared by Mann-Whitney U test. The relapse rate, overall survival (OS) and incidence of graft versus host disease (GVHD) were calculated by Kaplan-Meier curve.

RESULTSAll cases engrafted. 2-year OS rate was 86.0% (CI: 82.9% -89.1%), 2-year accumulated hematological relapse rate (RI) was 3.5% (CI: 1.7% -5.3%). 102 cases were not received any further intervention and none of them relapsed. The median value of bcr-abl transcripts was 25.800% (CI: 0.067% - 96.100%) pre-HSCT and 0.025% (CI: 0 - 3.583%) at +30 d, 0.011% (CI: 0-0.425%) at +60 d, 0.002% (CI: 0 - 0.610%) at +90 d, 0 (CI: 0 - 0.056%) at +120 d post HSCT. The bcr-abl mRNA levels decreased to undetectable level at +3 m, +4 m, and +5 m in recipients from HLA-mismatched related donor, identical sibling donor and unrelated donor respectively.

CONCLUSIONSSerial monitoring of bcr-abl transcript levels by real-time quantitative PCR after allo-HSCT is helpful for screening CML patients with low risk of relapse.

Adolescent ; Adult ; Child ; Female ; Follow-Up Studies ; Fusion Proteins, bcr-abl ; metabolism ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; therapy ; Male ; Middle Aged ; Postoperative Period ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult

Intracellular calcium overload is a key factor for myocardial ischemia reperfusion injury (IR). However, there was no report for interstitial calcium concentration dynamics. We investigated the interstitial calcium dynamics in rat myocardial IR model in vivo. A microdialysis system was involved, and the time delay of the system and recovery time was introduced and tested with a fluids switching method. Twelve SD rats were divided into IR or control group. Myocardial IR was induced by ligating (20 min) then releasing (60 min) the suture underlying left anterior descending branch. Mycrodialyisis probe was implanted into the left ventricular myocardium perfusion area for occlusion. Dialysate samples were collected every 10 min. Dialysate calcium concentration was detected with an atomic absorption spectrophotometer. Recovery time for the microdialysis system was 20 min, and recovery rate was 16%. Dialysate calcium concentration showed no changes during ischemia, descended immediately after reperfusion, reached the lowest level (67% of baseline value) 20 min after reperfusion, then escalated slowly. Recovery time was an important parameter for mycrodialysis technique, and it should not be neglected and needed to be tested. Our data suggest that interstitial calcium concentration in rats with myocardial IR in vivo kept steady in ischemia, descended rapidly at the initial reperfusion, then rebounded slowly. In conclusion, we introduced the concept of recovery time for microdialysis and provided a simple testing method.
Animals ; Calcium ; metabolism ; Dialysis Solutions ; metabolism ; Intracellular Space ; metabolism ; Kinetics ; Male ; Microdialysis ; methods ; Myocardial Reperfusion Injury ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spectrophotometry, Atomic ; Time Factors

OBJECTIVETo investigate the relationship between the polymorphic (AT)n repeats in 3ountranslated region of exon 4 of CTLA4 gene [CTLA4(AT)n] and Graveso disease (GD) in Zhuang nationality population of Guangxi province.

METHODSThe studied groups comprised 48 patients with GD and 44 normal controls. Amplification of target DNA was carried out by polymerase chain reaction (PCR). The amplified products were run by 8% polyacrylamide gel electrophoresis, and then followed by 0.1% silver staining. Some of amplified products were sequenced directly.

RESULTSNineteen alleles of CTLA4 gene microsatellite polymorphism were found in Guangxi Zhuang nationality individuals. The 106 bp long allele was apparently increased in patients with GD of Zhuang nationality but not in healthy controls (P< 0.05).

CONCLUSIONCTLA4 gene microsatellite polymorphism is strongly associated with Graveso disease in Zhuang nationality population of Guangxi province. CTLA4(AT)n 106 bp may be the susceptible gene in GD patients of Zhuang nationality in Guangxi; 19 alleles of CTLA4 gene microsatellite polymorphism were found in Guangxi Zhuang nationality individuals.

Adult ; Antigens, CD ; genetics ; Base Sequence ; CTLA-4 Antigen ; China ; Dinucleotide Repeats ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Graves Disease ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Sequence Analysis, DNA

OBJECTIVETo report an anatomical basis for the posterior tibial artery intermuscular septum branches island flaps and its clinical value.

METHODSThe origin,course, number, caliber and distribution of the posterior tibial artery intermuscular septum branches were studied in 10 adult cadavers(20 legs). 10 cases of homonymy and opposite side ankle and adjacent soft tissue defects with posterior tibial artery intermuscular septum branches island flaps, aged 20-50 years. Free skin transplantation on the donor sites.

RESULTSThe posterior tibial artery gives off 2-7 intermuscular septum branches. Its external diameter was about 0.4-1.8 mm,and the length was about 0.3-4.5 cm. The area of flaps was 7 cm x 6 cm - 20 cm x 8 cm in the group. All the flaps were survived except 1 with partial necrosis in the distant part(3 cm x 1 cm) of the flap. 7 cases were followed up two months to three years. The color, texture and appearance of the flaps were good.

CONCLUSIONSThe kind of flap has reliable blood supply, the scope of repairing was wide; avoidance of sacrificing the major artery; the flap was easy to be dissected. It was one of the ideal flaps to repair the ankle and adjacent soft tissue defects.

Adult ; Female ; Humans ; Male ; Middle Aged ; Surgical Flaps ; blood supply ; Tibial Arteries ; anatomy & histology ; transplantation ; Young Adult

ObjectiveTo observe the morphological changes of bone in the progress of chronic fluorosis.MethodsWistar rats were randomly divided into three groups,30 rats in each group:normal control group,experimental group Ⅰ and experimental group Ⅱ according to body weight.Rats in normal control group drank distilled water freely.Experimental group Ⅰ and group Ⅱ drunk distilled water with sodium fluoride preparation of fluorine containing ion 100,150 mg/L solution for six months,respectively.Bone mineral density was detected by X-ray,bone morphological changes were observed under light microscope and bone histomorphometric parameters were calculated using image analysis software.ResultsThe bone mineral density values were different statistically between the three groups after feeding for 2 and 4 months(F =19.79,3.28,all P ＜ 0.05).However no significant difference was found after feeding for 6 months(F =1.80,P ＞ 0.05).The bone mineral density of experimental group Ⅰ (0.20 ± 0.03,0.21 ± 0.03) was significantly higher than that of the normal control group(0.17 ± 0.03,0.20 ± 0.04) after feeding for 2 and 4 months.The bone mineral density of experimental group Ⅱ (0.21 ± 0.02) was lower than that of normal control group(0.22 ± 0.03) after feeding for 6 months.The bone lamella in experimental group Ⅰ was arranged disorderly,the number of osteocytes increased with their nucleus atrophy and the osteoblasts were more than that of control grouo which arranged in layers observed under light microscooy.In exoerimental group Ⅱ,the bone lamella was bent deformation,the number of osteocytes had decreased with their nucleus shrinking or even disappeared and the number of osteoclasts had increased significantly observed under light microscopy.In experimental group Ⅰ,the mean trabecular density [(0.33 ± 0.03)％] increased and the mean trabecular separation,thickness [( 163.57 ± 1.99),(59.26 ± 7.18 ) μm] decreased compared with that of normal control group [(0.31 ± 0.02)％,(186.60 ± 2.90)μm,(86.42 ± 1.48)μm,all P ＜ 0.05].In experimental group Ⅱ,the mean trabecular density[(0.26 ± 0.02)％] decreased,the mean trabecular thickness[(71.42 ± 10.77)μm] reduced compared with that of normal control group[(0.31 ± 0.02)％,(86.42 ± 1.48)μm].ConclusionsExcess fluoride can damage bone tissue.Low doses of fluoride can stimulate osteoblast activity and enhance osteogenesis.The activity of osteoblasts is great than that of osteoclasts.High doses of fluoride can stimulate both osteoblasts and osteoclasts activity,but mainly the activity of osteoclasts,and bone resorption increases.

Objective To study the genotyping distribution of the Yersinia pestis(Y.pestis)strains by characterizing the diversity of the insertion sequence IS100 within the Y.pestis genome.Methods Derived fromthe known sequence of oriental strain CO92,5 pairs of locus-specific primers originating from both sides of the adjacent region of IS100 copies were designed,and two other complementary primers inside the IS100 sequence were designed to correspond with the outer primers.Then,91 Y.pestis strains and l pseudotubebculosis strain were tested by the specific PCR method using the primers described above and the PCR products were conformed by the sequence analysis,then further analysis WaS performed after the IS100 status was marked on the map of the plague focus type of china.Results The 91 Y.pestis strains had different IS100 status in their genome on tested loci.some possessed IS100 insertion,some didn't,and others changed their genome constitution.The IS100 possession on the 5 loci also suggested a distribution of regionality.Conclusion The analysis of some IS100 insertion element loci reveals that the IS100 genotyping distribution is consistent with the plague focus of type of China.And IS100genotyping pattern of the Y.pestis stains well reflects its genome constitution and the high flowability in its natural evolution.