OBJECTIVETo observe the difference in the clinical efficacy on peripheral facial palsy between electro-acupuncture (EA) assisted with surface electromyography (sEMG) and conventional EA.

METHODSSixty cases of peripheral facial palsy were randomized into an observation group and a control group, 30 cases in each one. EA was applied during the first 15 days of sickness in the two groups, at Xiaguan (ST 7), Jiache (ST 6), Dicang (ST 4), Yangbai (GB 14), Taiyang (EX-HN 5), Quanliao (SI 18) and Hegu (LI 4), once a day. In the observation, group, 15 days after sickness, according to the situation in sEMG, on the basis of the acupoints in the previous treatment, the corresponding acupoints were reselected for EA. In the control group, the conventional EA was kept on. The treatment was given once every two days till the 35th day of sickness. Separately, on the 5th, 15th and 35th days of sickness, according to the detection of sEMG in the patients of two groups, the means ratios of: root mean square (RMS) of musculi buccinators, orbicularis oris, frontalis and nasalis on the healthy and affected sides were recorded and analyzed.

RESULTSThe differences of ratio in RMS of musculi buccinators, orbicularis oris, frontalis and nasalis on the healthy and affected sides were significant statistically in comparison between the, 15th day and the 5th day, and between the 35th day and the 15th day of sickness within each group (all P<0. 01). The differences of ratio in RMS of the muscles on the healthy and affected sides were significant statistically on the 15th and 35th days between the two groups (all P<0. 05).

CONCLUSIONEA assisted with sEMG achieves the significant efficacy on peripheral facial palsy, better than the conventional EA.

Acupuncture Points ; Adolescent ; Adult ; Aged ; Electroacupuncture ; Electromyography ; Facial Muscles ; physiopathology ; Facial Paralysis ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Objective To study the treatment effect by addition of granulocyte-colony-stimulating factor (G-CSF) that could reduce the level of residual disease in patients with Ph-positive chronic myeloid leukemia (CML) who appeared to have achieved a suboptimal response to imatinib (IM) alone. Methods Eleven patients with CML who had achieved≥ 35 % Ph-negativity on treatment of IM were enrolled. The initial dose of imatinib was 400 mg or 600 mg orally daily, and G-CSF, 5 μg/kg subcutaneously daily. The administration of G-CSF was postponed or interrupted in the event of leukocytosis (leukocytes ≥ 30 ×109/L) until the whitecell count fell ＜20 × 109/L. Efficacy was assessed by serial monitoring of blood levels of bcr-abl transcripts.Treatment with G-CSF was discontinued if the patient did not achieve a reduction in the transcript level of at least 0.5 log after 6 months. For patients whose bcr-abl transcript levels continued to decline but had not yet reached molecular remission, treatment was designed to continue for 1 to 6 months. Results Of 11 evaluable patients, nine had an appreciable decline in bcr-abl transcript levels(include 7 cases the reduction was greater than 1 log and 2 cases the reduction was greater than 0.5 log), 2 cases the reduction was lower than 0.5 log.In 7 cases the reduction was greater than 1 log, including five patients who did not achieved complete cytogenetic response and two patients achieved complete molecular responses. No bleeding episodes occurred.No patient discontinued therapy because of toxicity and there were no treatment-related deaths. Conclusion The addition of G-CSF should be considered safely and successfully for patients who fail to obtain optimal response to IM alone and this approach deserves further evaluation.

The present study retrospectively analyzed 2 patients suffering from refractory pure red cell aplasia after major ABO-incompatible hematopoietic stem cell transplantation who received treatment in the Henan Institute of Haematology between April 2004 and February 2006. Patient 1 was a 25-year-old female with acute lymphocytic leukemia in second remission, and patient 2 was a 16-year-old gid with acute myeloid leukaemia in second remission. The two patients received a transplant of human adipose tissue-dedved mesenchymal stem cells (1.0×106/kg). Both of them acquired rapid recovery from pure red cell aplasia without any side effects. These findings suggest that adipose tissue-dedved mesenchymal stem cells seem to be a promising therapeutic option in patients with refractory pure red cell aplasia after ABO-incompatible hematopoietic stem cell transplantation, in whom conventional treatment fails.

AIM: To study the effect of vascular endothelial growth factor(VEGF) on hematopoietic differentiation from mouse embryonic stem cells(ESC) in vitro.METHODS: ES-D3 was allowed to grow on mouse fetal fibroblast feeder layer,and then was developed into embryoid bodies(EB).EB cells were transferred into medium supplemented with different concentration of VEGF and VEGF+SCF for 1 week.Six groups,including.VEGF 5 ?g/L,VEGF 10 ?g/L,VEGF 20 ?g/L, VEGF 5 ?g/L+SCF,VEGF 10 ?g/L+SCF and VEGF 20 ?g/L+SCF,were designed.The group of spontaneous differentiation without cytokine(s) was used as control.Hematopoietic transcription factor GATA-2 and early hematopoietic differentiation genes(c-kit and ?-H1) were detected by RT-PCR.The content of CD34~+ cells in each group were measured by flow cytometry.The cells derived from ESC were incubated in semisolid methycellulose cultures.The numbers of total colony-forming units in culture(CFU-C) were counted by reverse microscope.RESULTS: ES-D3 grew and formed EB at day 4.VEGF had a stimulatory effect as a single factor on the expression of genes associated with early hematopoietic differentiation(GATA-2,c-kit and ?-H1),the generation of CD34~+ cells and CFU-C in EB.The effects of VEGF+SCF were the most potent in the experimental groups according to the percent of CD34~+ cells and the numbers of hematopoietic colonies.The most highest inducing efficacy was achieved in VEGF 20 ?g/L or VEGF 10 ?g/L combined with SCF.CONCLUSION: VEGF promotes the differentiation of ESC into hematopoietic cells in vitro.The strongest effect was achieved when VEGF was combined with SCF.

AIM: To investigate whether Flk1~+ CD31~- CD34~- cells isolated from human adult adipose tissue have characteristics of hemangioblasts in vivo. METHODS: After sublethally irradiated (300cGy) with a caesium source, the female non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice were injected with human adipose tissue-derived Flk1~+ CD31~- CD34~- cells (10~5 cells per mouse) via tail vain with 0.4 mL Roswell Park Memorial Institute medium (RPMI-1640). The control mice received the same volume of RPMI-1640 medium. All mice were killed 2 months after transplantation for further study. The differentiation potential of Flk1~+ CD31~- CD34~- cells was assessed in bone marrow and gastrointestinal tract by the methods of flow cytometry, RT-PCR, FISH, and triple-color immunofluorescence. RESULTS: Flk1~+ CD31~- CD34~- human adipose tissue-derived adult stem cells differentiated into endothelial cells and hematopoietic cells at the single-cell level in vivo. CONCLUSION: Human adult adipose tissue-derived Flk1~+ CD31~- CD34~- cells bear characteristics of hemangioblast in vivo and may have potential application for the treatment of hematopoietic and vascular diseases.

To assess the efficacy of cotransplantation of haploidentical mesenchymal stem calls (MSC) and hematopoietic stem cells (HSC) in the treatment of refractory severe aplastic anemia, Two child patients with refractory severe aplastic anemia admitted to Henan Institute of Haematology from August 2002 to December 2007 were selected. Adipose tissue-derived MSCs (AMSCs) were separately originated from haploidentical mother and peripheral blood stem calls (PBSCs) from HLA-identical sibling brother or sister of patients. The patient 1 received a cotransplantation of PBSCs and AMSCs (1 × 106/kg) at a dose of 4.5 × 108 mononuclear calls/kg (containing 4.41 × 106 CD34+ calls/kg and 0.11 ×105 CD3+ cells/kg); the patient 2 received a second PBSCT at a dose of 6.5 × 108mononuclear cells/kg (containing 4.62×106 CD34+ cells/kg and 0.12×105 CD3+ cells/kg) and AMSC (1 × 108/kg) from his haploidentical mother. The results show that the cotransplantation was successful. During the two years of follow up, the two patients exhibited good condition, with no other treatment or transfusion dependence.

BACKGROUND: The treatment of chronic myelogenous leukemia (CML) is revolutionized by the tyrosine kinase inhibitor imatinib mesylate (imatinib). However, resistance to imatinib is increasingly recognized as a clinical problem, the prognosis of patients who develop imatinib resistance is poor, particularly in acute transformation phase of leukemia.OBJECTIVE: To characterize a novel CML cell line and to further elucidate the mechanisms of resistance to STI571.DESIGN: An observational comparative experiment.SETTING: Henan Institute of Haematology, Henan Tumor Hospital.MATERIALS: Thirty female BALB/c nu/nu mice with 5 weeks old were purchased from Animal Center, Chinese Academy of Medical Sciences. STI571 was kindly provided by Novartis (Nuremberg, Germany). VP-16 was purchased from Bristol-Myers Squibb (Munich, Germany); anti-P-gp from Santa Cruz Company, USA; anti-ab1 from BD Biosciences Company, USA. The disposal of experimental animal was coincidence with the ethical standard.METHODS: The experiment was performed in the Henan Institute of Haematology from September 2003 to November 2005. A novel K562 cell line (K562/VP16) was achieved after exposure of the K562 cells to VP16. A small subpopulation (SP K562/VP16) that was capable of excluding Hoechst 33342 in the K562/VP16 cell line was isolated by flowcytometry sorting. The rest of the K562/VP16 cells were classified as non-SP K562/VP16. In order to elucidate the mechanisms involved in K562/VP16 SP cells which became resistant to STI571, the expression of multidrug-resistant gene 1 (MDR1), Bcr-Abl and P-gp was detected in K562, non-SP K562/VP16, or K562/VP16 SP calls, respectively. Furthermore, one thousand cells of K562, K562/VP16 SP and non-SP cells were injected,respectively, intraperitoneally into the right flanks of ten 5-week-old female BALB/c nu/nu mice. The same experiment was repeated twice.MAIN OUTCOME MEASURES: Comparison of STI571 resistance and oncogenicity of non-SP K562/VP16 and K562/VP16 SP cells.RESULTS: The MDR-1 gene expression of the Mr 170 000 P-gp was detected in K562/VP16 non-SP and K562/VP16 SP cells but not in K562 cells. The expression levels of P-gp in the two K562/VP16 cell lines were similar (P ＞ 0.05).The levels of Bcr-Abl and Abl proteins were similar in the K562 cell line and in non-SP K562/VP16 and K562/VP16 SP cells (P ＞ 0.05). Compared with non-SP K562/VP16, the K562/VP16 SP cells were more resistant to STI571, and this resistance could hardly be reversed by many multidrug resistance inhibitors. In addition, in vivo study showed that the K562/VP16 SP cells induced oncogenicity in mice, while the K562/VP16 non-SP cells failed to do so.CONCLUSION: Bcr/abl gene amplification and MDR1 overexpression may not be an important clinical mechanism in the diversity of resistance development to imatinib treatment, and the development of drug resistance by leukemia cells may be at least partly due to a rare SP cells which drives leukemia occurrence and maintenance. So, these SP cells need to be targeted for effective cancer therapy.

ObjectiveTo evaluate bacteria contamination during collection,processing and storage of cord blood to gain insight into contamination mechanism and direct prevention.MethodsFresh cord blood was separated by hydroxyethyl starch (HES) to harvest nucleated cells.The bacteria contamination was tested by culturing 10 ml plasma-red cells with BacT/ALERT 3D-480 automatic blood culture system.Total 87 positive samples were further identified for bacteria species.Ninety six cord blood nucleated cells concentrate with bacteria positive stored in liquid nitrogen(LN2) for 6-7 years were thawed at 37 C and re-cultured for bacteria analysis.ResultsWe collected 19 062 umbilical cord blood.Among them,336 was bacteria positive ( contamination rate 1.8 ％ ).Eighty-seven positive samples were further investigated with facultative bacteria 58 (66.7 ％ ),aerobic 38(43.7％ ) and anaerobic 17( 19.5％ ),Gram- negative accounted for 68％ while positive 32％.The most frequent bacteria were Escherichia coli ( 25.3％ ),Streptacoccus intermediate ( 14.9％ ) and Chromobacteria violaceum(9.2％ ).Ninety-six nucleated cells concentrate with bacteria positive were cryopreserved at liquid nitrogen for researching.Of them,83 samples( 86％ ) showed positive of bacteria culture after deep-low temperature storage for 6-7 years.ConclusionsBacteria contamination rate of the cord blood collection,processing and storage in 2000 ～ 2007 was 1.8％.Stored in liquid nitrogen for 6-7 years,the viability of bacteria was 86％.The aseptic procedures of cord blood collection in delivery room should be intensified.The bacteria re-culture following thawing of cord blood cells is necessary before clinical transfusion.

Objective To observe the efficacy and adverse events of L-asparaginasum plus DICE regimen in the treatment of relapsed and refractory non-Hodgkin's lymphoma (NHL). Methods Thirty-one patients with relapsed and refractory NHL were treated with L-asparaginasum plus DICE regimen. Each patient was scheduled to receive 2 to 6 cycles.Results Among the 31 assessable patients,11 (35.5 ％) achieved a complete remission (CR),14(45.2 ％) got a partial remission (PR),2 were stable,4 were progressive.The overall response (CR + PR) rate was 80.7 ％.The median survival was 8 months (rang:2-38 months).The 1-year survival rate was 43.3 ％,the 2-year survival rate was 32.5 ％.The main adverse events were myelosuppression,digestive tract reaction,allergy and edema.No treatment-related death was observed.Conclusion The L-asparaginasum plus DICE regimen is effective and safe for the relapsed and refractory NHL.

Objective To explore the efficacy and safety of moderate-dose of etoposide (VP16) with granulocyte-colony-stimulating factor (G-CSF) for mobilization of peripheral blood stem/progenitor cells.Methods VP16 at 1.2 g/m2 was injected intravenously by six divided doses via a central vein, 2 times every 12 hours for 3 days in 31 patients with malignant lymphoma (30 non-Hodgkin lymphoma and 1 Hodgkin lymphoma). All patients received G-CSF 5 μg/kg were given twice daily subcutaneously from the day of the nadir of white blood cell (WBC) till the day before the last APBSC harvest. Results The mean time for the collection of stem cell was 12 days (10-15) following etoposide chemotherapy. The mean number of mononuclear cell (MNC) and CD+34 cells in collection were 7.8×108/kg (5.2-11.3×108) and 7.2×106/kg (5.3-13.1×106). respectively. 18 patients completed collection with a single apheresis, and 13 patients underwenttwice. All patients were recovered for haematopoiesis in following APBSCT. Median (range) time for the recovery of absolute neutrophil count (ANC)＞0.5×109/L and platelet＞20×109/L were+12 (+9-+18) days and +14 (+10-+21) days respectively. Slight adverse events coursed by the regimen could be tolerated. Conclusion VP16 at moderate dose with G-CSF is an effective and safe mobilizing regimen for autologous peripheral blood stem/progenitor cells in patients with malignant lymphoma. It was suggested to use extensively.