Objective To isolate and identify leukemia stem cells from acute myeloid leukemia patients for further research. Methods By Ficoll density gradient centrifugation, mononuclear cells were firstly separated from bone marrow of patients. According to specific surface markers, CD+34 CD+123 of leukemic stem cells were sorted by flow cytometer. Their ability of self-renewal and differentiation were evaluated by colony formation and cobblestone forming ability. At the same time, the purity and cell morphology of CD+34 CD+123 cells was analysed. Results Comparared with total mononuclear cells, the proportion of the CD+34 CD+123 cells after sorting was 10.7 %, and these cells showed the ability of colony forming and cobblestone forming, and the purity proportion of CD+34 CD+123 cells was 62.1%. Conclusion The leukemia stem cells were isolated successfully and could be used in further study.

Objective To study the early and mid-term outcome in terms of left ventricular(LV)geometry and function in patients with transposition of great arteries with intact ventricular septum (TGA/IVS) undergoing arterial switch operation.Methods Twenty patients were followed up and divided into 2 groups according to age:the earlier group and the later group.Age and sexual matched controls were also designed as control group 1 and control group 2.LV posterior wall thickness(PWTd),dimension(LVIDd)and length(L) at end diastolic phase,LV end-diastolic volume (EDV),end-systolic volume (ESV),ejection fraction (EF),mitral valve early (E) and late (A) inflow velocities and E/A,isovolumic relaxation time (IVRT) were obtained by two-dimensional echocardiography,normalized isovolumic relaxation time (IVRTn),wall thickness index (PWTd/LVIDd) and LV geometry index(LVIDd/L) were calculated,and these indexes between groups were compared.Results ①Compared with control group 1,PWTd in earlier group was higher [(0.41 + 0.06)cm vs (0.36 + 0.05)cm,P =0.022],but LVIDd had no significant statistical difference [(2.08 + 0.21)cm vs (2.21 + 0.23)cm,P =0.117],indicating PWTd/ LVIDd differed between groups(0.20 + 0.04 vs 0.16 + 0.01 P =0.001).②No abnormalities were observed in later group.③Compared with different age groups in TGA,EDV,ESV,L,LVIDd,PWTd were significantly higher in later group(P =0.000).There were obvious correlations between EDV,ESV,L,LVIDd and examined age (r =0.90,0.83,0.73,0.90,P =0.000,R2 =0.809,0.694,0.528,0.808) while there were no significant correlation between PWTd and examined age(r =0.56,P =0.000,R2 =0.309).Conclusions Early after operation,LV is undergoing hypertrophy or hyperplasia in TGA/IVS,but will recover in the mid-term period,and the function is normal all the time indicating that the LV myocardium may avoided irreversible pathological changes if operated in time.All in all,LV develops well in mid-term period.

Objective:To study the clinical manifestations and treatments of closed injuries of the cervical trachea.Method:We carried out a retrospective study of the clinical manifestations, diagnosis and treatment of patients with closed injuries of the cervical trachea that have been treated in our hospital over the last ten years. We analyzed commonly occurring clinical problems, key points of surgical treatment, and postoperative recovery. Fourty-one patients with acute closed injuries of the cervical trachea underwent emergency tracheotomy or anesthesia orotracheal intubation and surgical repair of the trachea within 24 to 48 hours. Twenty six patients had their tracheotomy tubes extubated after surgery and had their anesthesia orotracheal tubes extubated within 48 hours postoperatively; tracheotomy tubes remained in the airway in 15 patients postoperatively, and of them, 12 had extubation successfully in 2 weeks postoperatively, while the remaining 3 were left intubated due to laryngotracheal stenosis. Among the 3, 2 underwent further surgical repair and recovery, and 1 had a stent inserted.Result:Twenty-eight patients fully recovered postoperatively. Ten patients mostly recovered postoperatively exception for hoarseness, and their fundamental phonation function recovered within 2 to 3 months but with poor movement of the vocal cords. The breathing and swallowing function of the other 3 patients recovered after the surgery, but they suffer from hoarseness.Conclusion:In the treatment of closed disruption of the cervical trachea, prompt diagnosis and timely surgical repair of the structure and function of the trachea are key to saving the patients' lives and avoiding tracheal stenoses.

Objective To observe the clinical efficacy ofXing Nao Kai Qiao(brain-awakening orifice-opening) needling method plus Frenkel's balance rehabilitation exercise in treating lower-limb dysfunction due to different types of ataxia after stroke.MethodA total of 115 eligible patients were randomized into two groups. Fifty-seven cases in the treatment group were intervened by Xing Nao Kai Qiaoneedling plus rehabilitation training; 58 cases in the control group were intervened byXing Nao Kai Qiaoneedling method alone. Berg Balance Scale (BBS) was adopted to evaluate the symptoms, balance function, and therapeutic efficacy before and after the treatment.Result In the treatment group, the markedly effective rate was respectively 72.2%, 83.3% and 61.1% in the cerebella subtype, brainstem subtype and basal ganglia subtype, and the total effectiverate was respectively 100.0%, 94.4% and 94.4%; in the control group, the markedly effective rate was respectively 21.1%, 35.0% and 58.8%, and the total effective rate was respectively 94.7%, 85.0% and 82.4%, and the between-group differences were statistically significant (P<0.05). After the treatment, the BBS score showed significant intra-group differences in both groups (P<0.01); the between-group comparisons showed that there were significant differences in comparing the scores of cerebella and brainstem subtypes after the treatment (P<0.05).ConclusionXing Nao Kai Qiaoneedling method plus Frenkel's rehabilitation exercise can effectively improve the symptoms of poststroke lower-limb ataxia, especially for cerebella and brainstem subtypes.

Objective:The effect and side effect of the dose-adjusted EPOCH regimen were evaluated perspectively for the pre-liminarily diagnosed angioimmunoblastic T-cell lymphoma. Methods: Nine cases of untreated angioimmunoblastic T-cell lymphoma were diagnosed and enrolled in our department from September 2008 to September 2012. All patients received dose-adjusted EPOCH regimen as first-line chemotherapy. Results: The median age of 9 patients was 54 years. The male-to-female ratio was 2∶1. About 88.9%of all patients were at Ann Arbor stageⅢ/Ⅳ, and 77.8%presented with B symptoms. Anemia was found in 66.7%of 9 patients, and lactate dehydrogenase elevated in 55.6%of patients. After an average of 4.7 cycles of chemotherapy of dose-adjusted EPOCH regi-men, the complete remission rate was 22.2%, and the total response rate was 66.7%. With a median follow-up of 20 months, the 4-year progression-free survival rate was 11.1%, and the overall survival rate was 33.3%. The median survival time was 19 months. The most common adverse events of EPOCH chemotherapy were hematologic toxicity. Grades 3-4 neutropenia and thrombocytopenia were re-ported in 77.8%and 33.3%of patients. Febrile neutropenia was observed in 44.4%of patients. Non-treatment-related mortality was al-so noted. Conclusion: The results of our research showed no clear benefit of treating preliminarily diagnosed angioimmunoblastic T-cell lymphoma with dose-adjusted EPOCH regimen. The main adverse events were hematologic toxicity and could be tolerated.

Objective To analyze the clinical characteristics and recent curative effect of mantle cell lymphoma (MCL) after conventional treatment.Methods Clinical data of 15 MCL patients admitted in the Affiliated Hospital of Academy of Military Medical Sciences between August 2004 and October 2013 were retrospectively analyzed.Results The median age of those patients was 59 and the male to female ratio was 1.5∶1.Fourteen(93%)cases were in Ann-Arbor stages Ⅲ -Ⅳ, 15 cases (100%)primarily with lymph node involvement,7 cases (47%)with bone marrow involvement,4 cases (27%)with gastrointestinal involvement,and 3 cases (20%)with orbit involvement.Less than 40% expression of Ki-67 was observed in 9 cases (60%),while 6 cases were with more than 40% (40%).One case was blastic variant.First-line therapy was CHOP-like regimens,which were combined with rituximab in 8 of the 15 cases.In this study,the median survival time was 12 months (3 -64),and the overal response rate was 80%after induction chemotherapy.The current survival of 7 /9 cases with less than 40% expression of Ki-67 was 8 -64 months,2 /6 cases with more than 40% expression of Ki-67 was 8 and 9 months,respectively.Conclusion MCL mostly occurs in older males.Extranodal invasion is common in MCL as an aggressive tumor.The efficacy of traditional chemotherapy is currently limited.Blastic variant or high expression of Ki-67 is an adverse prognostic indicator.

Objective To establish a convenient ~(13)C-breath test system in live mice,and investigate the value of ~(13)C-methacetin breath test(~(13)C-MBT) in the diagnosis of acute liver damage of mice with domestically synthesized ~(13)C-methacetin. Methods Domestically synthesized ~(13)C.methacetin was prepared from aeamol by methylation. Abdominal injection of CCl_4 was adopted to duplicate acute liver damage of mice,then the mice were housed under normal laboratory condition for a whole month to gain recovery,which were indentified by hepatic pathological examinations and biochemical teats of liver function.After fasting, the mice were orally administered ~(13)C-methacetin,and the expired air was collected at various time points. Infrared spectrometer was employed, and delm over baseline(DOB) curves of ~(13)C-exhalation were drawn. Results Six to eight min after administration of ~(13)C-methacetin,the rate of ~(13)C-exhalation peaked in control group(51.9±2.04), and decreased thereafter. Sixteen min after administration of ~(13)C-methacetin,the rate of ~(13)JC-exhalation peaked in model group(26.37±5.74), and decreased thereafter.There were significant differences between these two groups(P<0.05).There was no significant difference in peak value and time to reach the peak on DOB curves of ~(13)C-methacetin breath test after the two groups of mice were housed under the same condition for a month(P>0.05).Conclution ~(13)C-MBT facilitates the collection and evaluation of ~(13)CO_2 in the expired air of live mice,and yields precise reflection of alterations of liver function in acute liver injury and functional recovery.

OBJECTIVETo examine the feasibility of linking operons in tandem to enhance expression of heterologous genes in Escherichia coli (E. coli) and clarify the potential control mechanism of the total plasmid DNA amount in each host cell.

METHODSTwo series of expression plasmids, CW11 and CW12, containing 1 to 4 and 1 to 3 heterologous gene operon(s) respectively, were constructed. The molecular size of the CW11 series varied from 5.47 kb to 12.26 kb in 2.25 kb increments. The CW12 series varied from 5.40 kb to 9.72 kb in 2.16 kb increments. The expression level of desired protein was assayed by SDS-PAGE and laser density scanning. Plasmid copy number was determined by incorporation with (3)H-thymidine ((3)H-TdR).

RESULTSNo influence of the tandem-joined operons on host growth and plasmid stability was observed. Upon induction, the desired protein accumulations in the CW11 series were 44.9% +/- 3.9%, 51.3% +/- 4.1%, 54.8% +/- 3.3% and 58.2% +/- 3.4% of total cell protein. In the CW12 series, the yields were 32.2% +/- 5.0%, 42.8% +/- 4.1% and 46.9% +/- 4.0% of total cell protein. As size increased, the plasmid copy number decreased, but target gene dosage increased significantly (P < 0.01). Further calculation showed that the total amount of plasmid DNA per cell was not significantly different in each series (P > 0.05) and restricted to some extent.

CONCLUSIONSIncreasing the target gene dosage by tandem linking of operons may enhance the expression level of a desired protein. Although the size (kb) and the copy number of each plasmid are negatively interrelated, for certain plasmids in each series, their total DNA amount per cell seems to be a restricted constant for specific E. coli strains under identical incubation condition.

OBJECTIVE: To study the clinical manifestations and treatments of closed injuries of the cervical trachea. METHOD: We carried out a retrospective study of the clinical manifestations, diagnosis and treatment of patients with closed injuries of the cervical trachea that have been treated in our hospital over the last ten years. We analyzed commonly occurring clinical problems, key points of surgical treatment, and postoperative recovery. Fourty-one patients with acute closed injuries of the cervical trachea underwent emergency tracheotomy or anesthesia orotracheal intubation and surgical repair of the trachea within 24 to 48 hours. Twenty six patients had their tracheotomy tubes extubated after surgery and had their anesthesia orotracheal tubes extubated within 48 hours postoperatively; tracheotomy tubes remained in the airway in 15 patients postoperatively, and of them, 12 had extubation successfully in 2 weeks postoperatively, while the remaining 3 were left intubated due to laryngotracheal stenosis. Among the 3, 2 underwent further surgical repair and recovery, and 1 had a stent inserted. RESULT: Twenty-eight patients fully recovered postoperatively. Ten patients mostly recovered postoperatively exception for hoarseness, and their fundamental phonation function recovered within 2 to 3 months but with poor movement of the vocal cords. The breathing and swallowing function of the other 3 patients recovered after the surgery, but they suffer from hoarseness. CONCLUSION In the treatment of closed disruption of the cervical trachea, prompt diagnosis and timely surgical repair of the structure and function of the trachea are key to saving the patients' lives and avoiding tracheal stenoses.
Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Neck Injuries ; diagnosis ; surgery ; Retrospective Studies ; Rupture ; Trachea ; injuries ; Tracheal Stenosis ; Tracheotomy ; Young Adult

OBJECTIVE: To investigate the efficacy of round window catheter placement with dexamethasone perfusion for 13 patients with intractable sudden sensorineural hearing loss (SSNHL). METHOD: Dexamethasone (2.5 mg) was perfused transtympanically through round window catheter by Micro-infusion pump. The perfusion was taken twice a day and continued for an hour each time, for a total of 7 days. RESULT: Thirteen patients have been followed up for 1 month. Five of them (38.5%) were demonstrated a 17-54 dB improvement in pure-tone threshold average (PTA). The other 8 patients (61.5%) had no effect. CONCLUSION Round window catheter placement with dexamethasone perfusion is a cost-effective and useful treatment for SSNHL patients who had no effect through conventional therapy, although hearing thresholds of these patients had hardly improved to normal levels.
Adult ; Aged ; Catheterization ; Dexamethasone ; administration & dosage ; therapeutic use ; Female ; Hearing Loss, Sensorineural ; drug therapy ; Hearing Loss, Sudden ; drug therapy ; Humans ; Male ; Middle Aged ; Round Window, Ear