OBJECTIVETo investigate the effect of SIRT6/NF-κB signal axis in delaying hematopoietic stem/progenitor cell senescence with ginsenoside Rg1, in order to provide theatrical and experimental basis for looking for methods for delaying HSC senescence.

METHODSca-1 + HSC/HPC was isolated by magnetic cell sorting (MACS) and divided into five groups: the normal control group, the aging group, the positive control group, the Rg1 anti-senescence group, and the Rg1-treated group. Senescence-associated β-galactosidase (SA-β-Gal) staining, cell cycle analysis and hemopoietic progenitor cell mix (CFU-Mix) were adopted to determine the effect Rg1 in delaying or treating Sca-1 + HSC/HPC senescence biology. The mRNA and protein of senescence regulation molecules SIRT6 and NF-KB were examined by realtime fluorescence quantitative PCR (FQ-PCR) and western blotting.

RESULTCompared with the senescence group, the Rg1 anti-senescence group and the Rg1-treated group showed lower percentage in SA-β-Gal-stained positive cells, decreased cell proportion in G1 phase, increased number of CFU-Mix, up-regulated in SIRT6 mRNA and protein expression, down-regulation in NF-KB mRNA and protein expression. The Rg1 anti-senescence group showed more evident changes in indexes than the Rg1-treated group.

CONCLUSIONRg, may inhibit Sca-1 + HSC/HPC senescence induced by t-BHP by regulating SIRT6/NF-KB signal path.

Animals ; Antigens, Ly ; analysis ; Cellular Senescence ; drug effects ; Female ; Ginsenosides ; pharmacology ; Hematopoietic Stem Cells ; drug effects ; Male ; Membrane Proteins ; analysis ; Mice ; Mice, Inbred C57BL ; NF-kappa B ; physiology ; Signal Transduction ; physiology ; Sirtuins ; physiology

Objective: The current study was designed to distinguish Phase G0/G1， S， G2/M cell groups in the whole cell cycle by DNA flow cytometry. Method: The authors developed a multiparameter flow cytometry of cyclin E＋ A/DNA for MOLT-4. Result: The data showed that the method could distinguish Phase G0, early G1, late G1, S, G2,and M cells. The method could be used in molecular cell biology, especially in cell kinetic study.

OBJECTIVETo study clinical effects of one-hole microplate internal fixation for the treatment of collateral ligament injuries of the metacarpophalangeal joint of the thumb combined with fracture.

METHODSTwenty-two patients (16 males, 6 females) with collateral ligament injuries of the metacarpophalangeal joint of the thumb combined fracture were treated with one-hole microplate internal fixation. The age of the patients ranged from 18 to 53 years old with a mean age of 28.5 years old. The duration from injury to surgery ranged from 2 hours to 2 months, and the mean time was 6 days. All the patients had collateral ligament injuries combined with fracture of the metacarpophalangeal joint of the thumb. Thirteen patients had injuries in the right hand and 9 patients had injuries in the left hand. There were 18 cases of closed wound and 4 cases of open wound. Eighteen patients had fresh injuries (< 2 weeks) and 4 had old injuries (> 2 weeks). Sixteen patients had injuries in the ulnar collateral ligament of the thumb combined with fracture, 6 patients had radial collateral ligament injuries of the thumb combined with fracture, 4 cases of which were complicated with injuries of abductor pollicis brevis and the end of the flexor pollicis brevis tender. The size of the avulsed fragment was about 3.0 mm x 4.0 mm to 6.0 mm x 7.0 mm.

RESULTSThe incisions of 22 patients healed by first intention. The follow-up periods ranged from 6 months to 5 years old,with an average of 2.5 years old. The thumb function was evaluated by Saetta and other evaluation criteria, and 20 patients got an excellent result and 2 good.

CONCLUSIONThe application of one-hole microplate internal fixation in treating collateral ligament injuries with fracture of the metacarpophalangeal joint of the thumb is an effective method.

Adolescent ; Adult ; Bone Plates ; Collateral Ligaments ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Metacarpophalangeal Joint ; injuries ; surgery ; Middle Aged ; Thumb ; injuries ; surgery ; Young Adult

OBJECTIVETo investigate the influence of location and size of acute insular infarct on stroke-related electrocardiogram (ECG) changes and cardiovascular events.

METHODSNinety-nine cases admitted to hospital from October 2007 to June 2009, who were diagnosed as acute middle cerebral artery territory infarct within 48 h after onset and without the history of cardiac diseases, were included in the study. The patients were further divided into three groups: major insular infarct, minor insular infarct and control group, according to the infarct size on MRI diffusion-weighted image. The clinical data, ECG changes and cardiovascular events were compared between left and right insular infarct. Logistic regression was applied to determine the independent risk factors of ECG changes and cardiovascular events.

RESULTLarge artery atherosclerosis was the main cause of acute insular infarct (71.8 %), which was associated with higher NIHSS score compared to the control group (P < 0.01). Comparing the left and right insular infarct, the frequencies of sinus bradycardia and sudden cardiac death were significantly higher in left insular infarct (P < 0.01 and P < 0.05), while there was a trend that the frequency of atrial fibrillation was higher in right insular infarct (P = 0.079). With the larger size of insular infarct, the frequency of sinus bradycardia, new atrial fibrillation and sudden cardiac death (P<0.01, P<0.05 and P<0.05, respectively) became much higher. Logistic regression analysis showed that major insular infarct was related to the higher frequency of sinus bradycardia (OR = 4.660, 95% CI: 1.646 ~ 13.195; P = 0.004).

CONCLUSIONAcute insular infarct is associated with the stroke-related ECG changes and sudden cardiac death. Left insular infarct is related to sinus bradycardia, possibly due to the enhanced parasympathetic tone. It deserves clinical attention that the incidence of cardiac autonomic disturbance becomes higher with the enlarged insular infarct size.

Acute Disease ; Adult ; Aged ; Aged, 80 and over ; Brain Infarction ; complications ; physiopathology ; Death, Sudden, Cardiac ; etiology ; Electrocardiography ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Risk Factors

This study was aimed to investigate the expression of vascular endothelial growth factor (VEGF) in patients with aplastic anemia. The gene expressions of VEGF in mononuclear cells of bone marrow from 7 cases of aplastic anemia and 12 normal controls were detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The expressions of VEGF in bone marrow from 20 cases of aplastic anemia and 20 normal controls were also determined by immunohistochemistry assay. The results showed that expression of VEGF mRNA was found in 2 out of 7 (28.57%) bone marrow of patients and in 10 out of 12 (83.33%) bone marrow of normal controls. The VEGF mRNA in patients with aplastic anemia was significantly lower than that in normal controls (P < 0.05). No patients with aplastic anemia showed immunohistochemical staining of VEGF in bone marrow, while 5 out of 20 (25%) normal controls exhibited VEGF positive cells. Bone marrow of aplastic anemia patients contained less VEGF than that of normal persons (P < 0.05). In conclusion, when compared with normal controls, VEGF expression decreased significantly in patients with aplastic anemia at gene transcription level and protein translation level, it may be related to the defect of angiogenesis and thus hematopoiesis in bone marrow of patients with aplastic anemia.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; metabolism ; Base Sequence ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

Objective： This study was designed to establish a method of analyzing DNA content in fresh tumor using fresh cells. Methods： Clinical blood tumor samples were separated to obtain mononuclear cells by Ficoll-Hypaque, solid tumor tissues were made to be single cells by using mechanical method. The purified mononuclear cells and single cells suspensions were divided into two portions. One portion was fixed by ethanol and placed overnight, then measured DNA content by conventional flow cytometric method. The other portion was stained directly by PI （ propidium iodide) dissolved in PIPES (piperazine-Ν ,Ν '-bis[2-ethanesulfonic acid] disodium salt) buffer, then measured DNA content by flow cytometry, the DNA content results were compared with those of fixed cells. Results： There was no obvious difference in the CV（ coefficient of variation) of the peak of G0/G1 cells between fresh and fixed tumor cells from the same type tumor (t-test, P >0.05). Repetition test showed that the results of DNA content analysis using fresh cells were stable. Conclusion： Fresh tumor cells stained by PI dissolved in PIPES buffer could be used for the analysis of clinical tumor propagation and DNA ploidy.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Surgical Flaps ; Thumb ; surgery ; Toes

OBJECTIVETo investigate the value of real-time harmonic contrast-enhanced ultrasound (CEUS) in the diagnosis of atypical liver abscesses.

METHODSWe used Technos DU8 system with CEUS technique to examine 32 patients with suspected liver masses. Mechanical index was set at 0.08- 0.11. The contrast agent SonoVue was injected as a bolus (2.4 ml, < 5s) in the antecubital vein.

RESULTSA total of 36 lesions were studied in 32 patients, including 30 patients with single nodule and 2 patients with multiple nodules. All lesions with liver abscess were confirmed by biopsy. The whole-enhanced pattern in all lesions was clearly displayed in the arterial phase after the administration of SonoVue. The mean time of the beginning enhancement was (14.48 +/- 3.62) s, the mean peak enhancement time was (22.92 +/- 4.35) s, and the mean time of hypoechoic appearance of lesion was (80.30 +/- 35.30) s. The lesions with hypoechoic pattern (37.5%, 12/32) and isoechoic pattern (62.5%, 20/32) in the portal phase were detected by CEUS. The characteristic sign with honeycomb pattern on CEUS was shown in 30 (93.8%) liver abscess lesions.

CONCLUSIONCEUS is valuable in the diagnosis of atypical liver abscesses.

Contrast Media ; Humans ; Liver Abscess ; diagnostic imaging ; Ultrasonography

OBJECTIVETo evaluate the clinical and laboratory features of pediatric inv(16) acute myeloid leukemia (AML) retrospectively.

METHODDual color fluorescence in situ hybridization (D-FISH) using a LSI CBFβ inv(16) break apart probe labeled by Spectrum red and Spectrum green was performed in 15 acute myeloid leukemia cases, including 13 cases with or without abnormal eosinophils but with positive core binding factor β (CBFβ)-MYH11 fusion transcript detected by RT-PCR, and 2 cases with trisomy 8 (+8). The results were compared with the morphology, immunophenotype, karyotype and RT-PCR.

RESULTMorphologically, 12 cases were diagnosed as M(4)EO, 2 as M(4), and 1 as M(2a). Immunophenotypically, all 13 AML cases with inv(16) showed positive expression of CD(13) and CD(33), but without the expression of any lymphoid lineage antigens. Karyotyping analysis with G-banding detected inv(16) in 10 AML cases, including 9 M(4)EO cases and 1 M(2a), but only 5 positive cases were detected using R-banding technique. Among them, 2 cases had simultaneous +8 and trisomy22 (+22), one had +22 only in addition to inv(16). D-FISH revealed a CBFβ-MYH11 rearrangement in 13 cases of AML with positive RT-PCR results, and the mean positive rate of cell detection was 55.15% (range 37.0% - 86.0%). The complete remission rate (CR) and median survival period in this series of inv(16) AML were 81.5%and 11 months, respectively, of whom, 8 cases were still in CR. Relapse and karyotypic evolution were seen in case 5 with +8, +22 in addition to inv(16).

CONCLUSIONAML with inv(16) is a special subtype. Most cases belong to M(4)EO. Its prognosis is good in general, but it seems to be an unfavorable feature for AML with inv(16) and +8, +22 simultaneously, especially with karyotypic evolution. For detection of inv(16), G-banding technique is evidently superior to R-banding technique. D-FISH combined with RT-PCR are more sensitive and reliable than chromosome banding analysis.

Adolescent ; Child ; Child, Preschool ; Chromosome Deletion ; Chromosome Inversion ; Chromosomes, Human, Pair 16 ; genetics ; Eosinophilia ; pathology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Male ; Prognosis ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the risk factors of mortality in patients with severe chest trauma (SCT).

METHODSThe clinical data of 777 SCT [abbreviated injury scale (AIS) ≥3] patients who were treated in the Chongqing Emergency Medical Center from January 2006 to April 2009 were retrospectively reviewed. Stepwise logistic regression analysis was used to explore 15 possible mortality-related risk factors.

RESULTSSeven factors were found to be correlated with the mortality of SCT: age, hemorrhagic shock, multiple organ dysfunction syndrome (MODS), pulmonary infection, abdominal organ injury, Glasgow coma scale (GCS) score, and thorax AIS score. Among them five factors were the independent factors that might increase the mortality of SCT: hemorrhagic shock (B=1.710, OR=1.291, P=0.001), MODS (B=3.453, OR=1.028, P<0.001), pulmonary infection (B=2.396, OR=10.941, P<0.001), abdominal organ injury (B=1.542, OR=1.210, P=0.005), and thorax AIS score ≥4 (B=0.487, OR=1.622, P<0.001). Two factors showed protective effects: age ≤60 years (B=-0.035, OR=0.962, P=0.01) and GCS score ≥12 (B=-0.635, OR=0.320, P<0.001).

CONCLUSIONSAge, disease severity, and complications (hemorrhagic shock, MODS, and pulmonary infection) are independent risk factors of the mortality of SCT. Effective treatment programs targeting these risk factors may improve the outcomes of SCT patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Thoracic Injuries ; mortality ; Young Adult