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.

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.

BACKGROUND: There is no consistently effective therapy for patients with steroid-refractory acute graft-versus-host disease (GVHD). A variety of alternative approaches have been tested, including antithymocyte globulin, mycophenolate mofetil (MMF), pentostatin, and monoclonal antibodies; however, these treatments have been only modestly successful. OBJECTIVE: To further evaluate the efficacy of human adipose-derived stem cells (ASCs) as the salvage therapy for steroid-refractory acute GVHD. DESIGN: A clinical trial.SETTING: Department of Haematology, Henan Institute of Haematology, Henan Tumor Hospital.PARTICIPANTS: The clinical trial was performed at the Henan Institute of Haematology from September 2002 to August 2005. Eight patients were treated with ASCs for grades Ⅲ-Ⅳ steroid-resistant acute GVHD. The study was approved by the Ethics Committee at Henan Tumor Hospital and informed consent was obtained from patients and ASCs donors before they enrolled.METHODS: Eight patients with steroid-refractory grades Ⅲ-Ⅳ acute GVHD received intravenous infusions of ASCs. The ASCs dose was 1.0×106/kg. Seven patients were treated once and one patients twice. Four patients received ASCs from haplo-identical family donors and four from unrelated mismatched donors. MAIN OUTCOME MEASURES: The efficacy of human ASCs as the salvage therapy for steroid-refractory acute GVHD. RESULTS: No side effects were noted after the ASCs infusions. Acute GVHD disappeared completely in seven of eight patients and six of these seven patients are still alive after the median follow-up of 30 months (range 11-90 months) after the initiation of ASCs therapy. All four surviving patients were in good clinical condition and in remission of their hematological malignancy. Two patients died-one with no obvious response to ASCs died of multiorgan failure and one of relapse of leukemia. CONCLUSION: These results suggest that ASCs is a very promising treatment for severe steroid-resistant acute GVHD.

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.

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.

Objective To explore the feasibility of establishment of NOD/SCID-mouse model with multiple myeloma by using plasma cells from myeloma patients.Methods The femurs and tibias were removed from the New Zealand white rabbits;the muscles,periosteum and cartilage tissues were cleared.Then each bone was cut into two pieces gently along its middle.The NOD/SCID mice weighing 25 - 30 g (4 - 6 weeks)were anesthetized by intraperitoneal injection;rabbit bone was inserted into the right side of the mouse back and engraftment of the bones was allowed to take place after 4 weeks.The 5000 000 purified plasma cells which expressed CD38 +/CD45 - were immunofluorescence labeled and then injected slowly into the implanted rabbit bone through the distal end.The mice were observed weekly;the plasma cells growth in mice was screened by the living-imaging system and the tumor from the mice was determined by biopsy.Results The implanted rabbit bone survived after 4 weeks.The tumor in mice was observed 2 weeks after the purified myeloma cells were injected into the rabbit bone,and it reached 100 mm3 after 8 weeks.Results of the living-imaging system showed that the myeloma cells had uptake in the rabbit bone after 2 weeks of injection and this phenomenon was more pronounced after 8 weeks of injection (2.4×10 4 vs .1.5× 10 5 ,P < 0.05 ).The tumor infiltrated with numerous plasma cells and osteoclasts increased upon the biopsy. Conclusion Rabbit bone marrow implanted into NOD/SCID mice can effectively support local injection of plasma cells of multiple myeloma patients,and the NOD/SCID-mouse model of myeloma has been established.This model can be used to study in vivo experiments related to myeloma and clinical therapeutic approaches for this disease.

Purpose To investigate the expression of C-C chemokine receptor 9 (CCR9) in non-small cell lung cancer (NSCLC) and to explore its prognostic value. Methods The expression of CCR9 was detected by immunohistochemistry in tumor tissues and corre-sponding adjacent normal tissues from 119 NSCLC patients. Additionally, the correlation between CCR9 expression and the clinicopath-ologic features of NSCLC and the relationship between prognostic factors and overall survival rate were analyzed by statistical methods. Results The positive expression rate of CCR9 was significantly higher in NSCLC tissues (54. 6%) than that in adjacent normal lung tissues (10. 1%) (P<0. 05). The expression of CCR9 in NSCLC was correlated with histopathologic type, lymph node status and p-TNM stage (P<0. 05). Kaplan-Meier survival analysis suggested that the positive expression of CCR9 was negatively correlated with the overall survival rate (Log-rank=9. 917, P=0. 002). Univariate analysis showed that the lymph node status, p-TNM stage and the positive expression of CCR9 made great difference to postoperative overall survival (P<0. 05). Multivariate analysis showed that CCR9 expression was an independent prognostic factor for overall survival of NSCLC patients ( RR=0. 447, 95%CI:0. 201 ~0. 993, P<0. 05). Conclusion The expression of CCR9 may predict a poor prognosis in the patients with NSCLC, so it can be used as a novel NSCLC biomarker.

Objective To investigate the clinical characterstics of bone Langerhans-cell histiocytosis (LCH) and evaluate its diagnosis,therapy and prognosis.Methods 25 cases with biopsy confirmed bone LCH during the last 8 years were retrospectively analyzed.Results The patients included 18 males and 7 females,13 children and 12 adults,ranging from 1.5 to 55 years old with a median age of 17.Cases with unifocal lesions were 17,including 11 cases of skull LCH,and the remaining 8 were with multifocal lesions.First symptoms were predominantly pain and local masses,and rarely constitutional symptoms.The manifestation of radiography was osteolytic bony lesions.12 cases had masses in soft tissues.Patients with solitary lesions underwent surgical operation,followed by radiotherapy or chemotherapy.Cases with multifocal lesions received chemotherapy and radiotherapy.Pathological examination showed proliferation of well differentiated histiocytes,and large numbers of infiltrating eosinophils.Positive rates of CD1a,S100,Vimentin and CD68 were higher in immunohistochemistry.Patients with restricted involvement in bones can achieve a satisfactory therapeutic effect.2 cases died when multiple systems were compromised.Conclusion Bone LCH occurs predominantly in children and teenagers,involves solitary bones,and morbidities in males are much higher than females.Skull is most commonly affected.Principal clinical manifestations are pain and local masses.Diagnosis of bone LCH depends on microscopic examination.Combination therapy appears to be an effective method of treatment.Prognosis of disease is related to the degree of bone involvement,histological classification and simultaneously encroachment of other organs.Most patients have good prognosis.