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This study was to investigate the relationship between Clostridium difficile associated diarrhea (CDAD) and intestinal microecosystem in patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT) and to clarify clinical characteristics of intestinal microecosystem disorder. Clostridium difficile (CD) was isolated and identified by enzyme-linked-immunosorbent assay using clostridium difficile Premier toxins A&B Kit and anaerobic culture in 44 cases with diarrhea. Fecal flora (bifidobacteria, lactobacillus, bacteroides, peptostreptococcus, Clostridium perfringens, enterobacteriaceae, enterococcus, and yeasts) of patients were quantitatively and qualitatively analyzed by Mitsuoka's methods. The results showed that CDAD occurred after using antibiotic or chemotherapy. Clostridium difficile was detected in 12 patients with diarrhea (positive rate was 27.27%). There was marked changes of intestinal microecosystem when patients suffered from CDAD. The number of lactobacillus, bifidobacteria, bacteroides, enterobacteriaceae and so on decreased significantly. It was effective to treat CDAD with vancomycin, metronidazole and probiotic, but the recurrence rate was 16.67%. In conclusion, CDAD complicated by allo-HSCT is related to change of intestinal microecosystem. While treating CDAD with the sensitive antibiotic, the intestinal flora of patients should be supported actively. This treatment contributes to improving disease status and reducing diarrhea recurrence.

The aim of this study was to investigate the expanding capacity of bone marrow-derived mesenchymal stem cells (MSCs) in 34 patients who received a marrow and/or peripheral blood stem cells transplant (SCT). Marrow samples were obtained from iliac crest aspirates of healthy individuals (normal controls) and patients for the isolation, purification, and expansion of MSCs. The different passage MSCs of patients were analyzed by flow cytometry (FCM) to reveal the cell surface antigen expression. The expanding function of MSCs from patients after SCT, which might be affected by cytotoxic therapy in the conditioning regimen, colony-forming unit-fibroblast (CFU-F), confluent time, and passage number of the culture were measured, and then compared with those in the normal controls. At the same time, the numbers of colony-forming unit of hematopoietic progenitor were detected and compared with normal controls. In addition, CFU-F, confluent time, and passage number of MSCs were compared between group of BMSCs plus PBSCs co-transplanted patients and group of BMSCs plus PBSCs with the second donor umbilical cord blood cells (UBCs) co-transplanted patients. The results indicated that a confluent monolayer of stroma cells was generated in 31 out of the 34 cases (91.1%), a subconfluent monolayer was generated in one case (2.9%), no adherent stromal layer was generated in 2 cases (5.9%). As compared with the normal controls, the time generating a confluent layer of stroma cells from primary MSCs of patients was longer significantly, and the passage number and CFU-F of MSCs of patients in vitro was less than that of normal controls significantly. Compared with the group of BMSCs plus PBSCs co-transplanted patients, the confluent time of MSCs in group of BMSCs plus PBMSCs with UBCs co-transplanted patients was shorter, the passage number and CFU-F count in this group were higher. It is concluded that the MSCs of patients after HSCT are damaged, and the co-transplant of BMSCs and PBSCs plus UBCs can partially improve in vitro expanding capacity of MSCs from patients.

To investigate the cytogenetic and clinical characteristics of inv(3q) (q21q26) and t(3;3) (q21; q26) aberrations as well as prognosis, cases were collected and chromosome specimens of bone marrow cells were prepared by 24-hour culture, while G-banding technique was used to perform karyotyping. The results showed that the simple inv(3q) and t(3; 3) aberrations were rare, they commonly combined with other chromosome aberrations such as -7/7q- and t (9; 22). The involved diseases included myelodysplastic syndromes, acute myeloid leukemia and chronic myelogenous leukemia in blast crisis. Out of 24 patients, 2 patients diagnosed with M(5) subtype did not achieve complete remission after multiple chemotherapy; 2 patients received allogenic stem cell transplantation relapsed. It is concluded that 3q21q26 aberration commonly combined with chromosome aberration 7/7q-, for these patients the efficacy of chemical therapy is poor, the efficacy of bone marrow transplant is too poor, these patients with inv(3q) and t(3; 3) aberrations have poor prognosis and short overall survival.

LI.4 is a major acupoint but the method of locating it has not been standardized. In fact, description of method for locating this acupoint often varies in the classic and traditional texts. It might signify this point may be varied from one to another person. Our comparative study of locating and subsequent acupuncturing these locations revealed some interesting features of LI.4 from our collected clinical data in that location 1 had a better therapeutic effect for toothache relief and analgesia-anesthesia effect than for headache relief, location 2 was better for headache than toothache relief; locations 3 and 4 had a mixed effect whereas, location 5 seems to have a better chance to get bioenergy (De Qi) as manifested by tingling and numbing sensation at the acupunctured site. Our study also suggested that LI.4 location was about the size of 4 millimeters in diameter instead of a pin-point location, though needle inserted at correct point produced a better result.

Objective: To explore the immunophenotypic characteristics of CD7 and/or CD56 positive acute myeloid leukemic stem cells, and the relationship between minimal residual disease (MRD) and the leukemic stem cells (LSC)., Methods: The immunophenotype of leukemia cells from 51 CD34+ CD38+ CD7+ and/or CD34+ CD38+ CD56+ acute myeloid leukemia (AML) patients (exclude M3) at diagnosis was analyzed by using 4 - 6 panels of 4 color antibodies, and cells from 28 normal bone marrow (NBM) samples were served as control. The expression of CD7 and CD56 in the CD34+ CD38+ subpopulation was used as a leukemic cell marker for monitoring MRD of 53 samples from 26 CD7+ and (or) CD56+ patients., Results: In CD7+ and/or CD56+ AML patients at diagnosis, the average positivity of CD7 in CD34+ CD38+ subpopulation and CD34+ CD38- Lin- stem cells subpopulation was (77.39 +/- 20.71)% and (44.57 +/- 22.70)%, and that of CD56 was (56.71 +/- 32.56)% and (33.51 +/- 29.64)%, respectively, all significantly higher than that of NBM (P < 0.01 and P < 0.05). Compared with that of NBM, the expression of CD90 in AML patients was significantly lower in the CD34+ CD38- Lin- subpopulation (P < 0.01), the expression of CD123 was significantly higher than NBM (P < 0.01), and the expression of CD117 was no significant difference (P > 0.05). In follow up of CD7+ and (or) CD56+ patients, the expression rate of CD7 and (or) CD56 in the CD34+ CD38- Lin- subpopulation MRD+ group was significantly higher than that in the MRD- group. The actual rate for CD7 was 71% (15/21) and 16% (4/25) (P = 0.001), and its relative rate was 81% (17/ 21) and 24% (6/25) (P = 0.000), respectively. The actual rate of CD56 is 100% (4/4) and 12% (3/25) (P = 0.001), and its relative rate was 75% (3/4) and 20% (5/25) (P = 0.031), respectively. A high CD7+ CD34+ CD38- Lin- subpopulation frequency at diagnosis in CD7+ AML patients predicted a high frequency of positive MRD in later detection (P < 0.05)., Conclusions: CD7 and CD56 are expressed on the stem cells in CD7+ and/or CD56+ AMLs and a high frequency of CD7 and CD56 in the CD34+ CD38- Lin- stem cell subpopulation predicts a high frequency of positive MRD in later detection.

Objective: To report two cases of glandular odontogenic cyst and examine its cytokeratin 18,19 expression., Methods: Two cases of glandular odontogenic cyst were reported and studied. The cytokeratin 18, 19 expression in these two cases were also investigated using immuno-histochemical staining as well as in the situ hybridization of the cyst epithelium., Results: Histo-pathological examination revealed that ciliated columnar cells, squamous cells and low-columnar cells were found in the superficial layer of the lining epithelium. Several minor salivary glands, mainly composed of seromucous cells were observed near the satellite cyst. CK18 were expressed in all layers of the lining epithelium of varying intensity. CK18 was negative in lining epithelium of the daughter cyst, but CK19 was positive. CK18-mRNA was expressed in all the layers of the lining epithelium, the salivary glands and daughter cysts., Conclusions: Histological features and CK18 expression may be indicative of the possibility of salivary glandular and odontogenic differentiation.

Haploidentical transplantation is a feasible alternative for patients with life-threatening hematologic diseases who lack a matched donor. Factors affecting the clinical outcomes of haploidentical transplantation remain under investigation. We analyzed 157 consecutive patients with leukemia who underwent transplantation with nonmanipulated granulocyte colony-stimulating factor (G-CSF)-mobilized marrow and peripheral blood cells (G-BMPBs) from haploidentical donors after receiving myeloablative chemotherapy (Ara-C + BuCy + antithymocyte globulin). Follow up observations after transplantation were made from 48 days to 1191 days (median, 448 days). Multivariate analysis indicated that the cohort given higher doses of CD3(+) cells (> or = 177 x 10(6) /kg) in allograft transplantation had a significantly lower treatment-related mortality (TRM) (relative risk [RR] = 0.35; 95% CI = 0.16-0.77; P = .0090), better leukemia-free survival (LFS) (RR = 0.46; 95% CI = 0.26-0.84; P = .0106), and better overall survival (OS) (RR = 0.42; 95% CI = 0.23-0.78; P = .0058). Inversely, advanced-stage disease was a strong predictor of greater posttransplantation relapse (RR = 3.48; 95% CI = 1.26- 9.60; P = .0159), worse LFS (RR = 2.56; 95% CI = 1.33-4.95; P = .0050), and worse OS (RR = 2.77; 95% CI = 1.39-5.53; P = .0038). A high number of CD3(+) cells (> 177 x 10(6)/kg) given to patients resulted in statistically less TRM and more intensive graft versus leukemia effect without producing more severe grades of GVHD, all resulting in a significantly better overall clinical outcome from haploidentical transplantation.

Objective: To retrospectively analyze the results of a consecutive series of 90 refractory/relapsed acute leukemia (AL) patients receiving allogeneic hematopoietic stem cell transplantation (allo-HSCT) in our center., Methods: Of the 90 refractory/relapsed AL patients, 56 were male and 34 female, with a median age of 37 (13 - 59) years. Among them, 73 patients suffered from AL including 11 Ph+ acute lymphoid leukemia in first complete remission, 23 in second or greater complete remission, 39 in non-remission or relapse and 17 patients suffered from myelodysplastic syndrome (MDS-RAEB or RAEB-T) before transplant. Allo-HSCT from HLA identical siblings was performed for all the patients, of whom 27 received bone marrow transplantation (BMT), 30 peripheral blood stem cell transplantation (PBSCT) and 33 BMT + PBSCT. Eleven patients underwent allo-HSCT with conditioning regimen of CY/TBI and 79 with BU/CY. CsA + MTX regimen was use for prophylaxis of graft-versus-host disease (GVHD). The average follow-up was 15 months., Results: At the last follow-up, 56/90 (62.2%) survived, 50/90 (55.5%) survived without leukemia and 28/90 (31.1%) relapsed. The estimated 4-year overall survival (OS) and disease-free survival (DFS) of the 90 cases was 45.5% and 34.9%. The 4-year OS and DFS were significantly higher for patients in CR (54.0%) or MDS (70.1%) than in non-remission and relapsed (28.2%) patients before allo-HSCT (P = 0.027). The outcome of grade 0-I acute GVHD was better than that of II-IV GVHD (57.6% vs 26.7%, P = 0.015). Sex, age, central nervous system leukemia, conditioning regimen and the source of stem cell were not the factors affecting OS, DFS, cumulative incidences of relapse rate and treatment related mortality. Multivariate analysis showed the most significant factor associated with high DFS was the disease state., Conclusions: allo-HSCT can cure a significant proportion of refractory/relapsed AL patients, especially for those in CR. Early allo-HSCT is recommended for MDS patients.

Objectives: To evaluate the efficacy and safety of imatinib mesylate (imatinib) for patients with Philadelphia chromosome-positive (Ph+ ) chronic myeloid leukemia (CML) in accelerated and blastic phase., Methods: Seventy-five Ph+ CML patients in accelerated phase and 49 in blastic phase were treated with 400 mg or 600 mg of imatinib once daily., Results: For patients in accelerated phase, the cumulative hematological response (HR) rate was 93.3%, including complete HR (CHR) rate 85.3%, and returning to chronic phase (RCP) rate 8% in a median follow-up of 23.0 (1.0 -64.0 ) months. Cumulative major cytogenetic response (MCyR) rate was 33.0%, and complete cytogenetic response (CCyR) rate 28.0%. For patients with CCyR, the major molecular response (MMoR) rate was 47.6%. The estimated 4-year progression-free survival (PFS) rate and overall survival (OS) rate were 48.2% and 52.2% in patients with HR, respectively. Severe leukocytopenia, anemia and thrombocytopenia occurred in 37.3%, 34.6% and 45.3% of all patients, respectively. For patients in blastic phase, the cumulative HR rate was 63.3%, including CHR rate 44.9%, and RCP rate 18.4% in a median follow-up of 4.5 (0.3 -63.0) months. Cumulative MCyR rate and CCyR rate were both 12.2%. For patients with CCyR, the MMoR rate was 33.3%. For patients with HR, the estimated 1-year/2-year PFS and OS rates were 32.8%/15.8% and 46.0%/ 21.0% respectively. Severe leukocytopenia, anemia and thrombocytopenia occurred in 75.5%, 71.4% and 73.5% of all patients, respectively., Conclusions: The efficiency of imatinib was decreasing, and severer hematological toxicities increasing with the disease progressing in patients with Ph+ CML. Imatinib improves progression-free survival significantly in most patients in accelerated phase, particularly in those with continuous CCyR or MMoR. The response duration in majority of blastic phase patients is short, and the relapse rate is high.

Background: Real-time quantitative RT-PCR (RQ-PCR) assay has become a vital tool to monitor residual disease of leukemia. However, the complexity and standardization of RQ-PCR should never be overlooked and the results should be interpreted cautiously in clinical conditions. We aimed to assess the methodology of RQ-PCR and its clinical applications in monitoring molecular kinetics of 36 newly diagnosed cases of acute promyelocytic leukemia patients with t (15; 17) from October 2004 to December 2005., Methods: All the TaqMan probe-based RQ-PCR reactions and analysis were performed on an ABI-PRISM 7,500 platform. The quantitation of PML-RARalpha transcripts was represented by the normalized quotient, that is, PML-RARalpha transcript copies divided by ABL transcript copies. According to induction therapy, the patients were classed into two groups: group 1 (n = 23), three-drug combination including arsenics, all-trans retinoic acid and mitoxantrone; and group 2 (n = 13), two-drug combination from all-trans retinoic acid, arsenics and mitoxantrone., Results: The sensitivity of RQ-PCR was 1 per 10(5) cells and 5 copies of the PML-RARalpha transcript could be reproducibly detected. No false positive results occurred in 40 non-acute promyelocytic leukemia samples. Optimal amplification efficiency could be attained, which was determined by the slope of the standard curves (slope: -3.2 - -3.7). The inter-assay and intra-assay variation coefficients of the method were 1.01% and 0.56% respectively. Although the time to attain hematological complete remission was similar in both groups, the time to achieve molecular remission of group 1 was significantly shorter than that of group 2 (61 days vs 75 days, P = 0.034). The rate of molecular remission within 70 days was higher in group 1 than in group 2 (75.00% vs 38.46%, P = 0.036). Compared with pretreatment, median reduction of the PML-RARalpha transcript before first consolidation therapy differed significantly between group 1 and group 2 (log scale, 3.15 vs 2.31, P = 0.024). Interestingly, we found that PML-RARalpha transcript levels temporarily increased in bone marrow (7 patients) and peripheral blood (22 patients) samples of patients during induction therapy in both groups., Conclusions: The RQ-PCR assay is reliable for the detection of PML-RARalpha transcripts. Arsenics, all-trans retinoic acid and mitoxantrone triad induction treatment of acute promyelocytic leukemia is superior to two-drug combination induction therapy in terms of the molecular response.

Background: The definite pathogenesis of hemorrhagic cystitis (HC) after allogenic hematopoietic stem cell transplantation (allo-HSCT) has not been well elucidated. The role of cytomegalovirus (CMV) reactivation and graft-versus-host disease (GVHD) in the development of HC remains obscure. This study determined the incidence and risk factors for HC after allo-HSCT and analyzed its association with CMV reactivation and GVHD., Methods: We retrospectively studied 250 patients at high risk for CMV disease who underwent allo-HSCT all based on busulfan/cyclophosphamide (BU/CY) myloablative regimens. The incidence, etiology, risk factors and clinical management of HC were investigated., Results: HC developed within 180 days of transplant in 72 patients, with an overall incidence of 28.8% and an incidence of 12.6% in patients with HLA-matched related donors (MRD), 34.38% in those with HLA-matched unrelated donors (MUD), 49.45% in those with mismatched related donors (MMRD). CMV-viremia significantly increased the incidence of later onset HC (LOHC); however, only 9 out of 15 patients with CMV viruria actually developed LOHC. Multiple regression analysis identified grade II - IV acute GVHD (RR = 2.75; 95% CI 1.63 +/- 4.66; P < 0.01) and grafts from MUD or MMRD (RR = 2.60; 95% CI 1.52 +/- 5.20; P < 0.01) as independent risk factors for HC. Event sequence analysis indicated a majority of HC episodes began around GVHD initiation., Conclusions: CMV-viremia is a high risk factor for LOHC. Our data also showed a correlation between acute GVHD and HC, which suggested that alloimmunity may be involved in the pathogenesis of HC.

This article describes a case of persistent saliva drooling, and the accompanying chronic angular cheilitis, that is not uncommon in the elderly patient population. A drug therapy and the temporomandibular joint aspect of vertical dimension of occlusion during prosthodontic evaluation and construction are presented. Also described is a method to incorporate a canula into the denture prosthesis to channel the saliva toward the oropharyngeal area for geriatric and handicapped patients who suffer from chronic drooling and angular cheilitis.

Objective: To study the correlation between polyoma virus load and hemorrhagic cystitis after allogeneic stem cells transplantation for prevention of hemorrhagic cystitis., Methods: Blood and urine specimens were collected from 40 healthy persons, 40 patient with stem cells transplantation and 20 cases complicated with hemorrhagic cystitis for determination of VP1 gene of polyomaviruses BK virus (BKV)/Jamestown Canyon virus (JCV) and simian virus 40 (SV40) by polymerase chain reaction (PCR) and EvaGreen stain fluorescence quantitative assay., Results: In the peripheral blood, all genes of BKV/JCV and SV40 were negative, while BKV gene in urine and blood from healthy persons and patient with stem cells transplantation was 15% (6/40) and 100% (40/40), respectively. The gene of JCV was positive in 10% (4/40) and 12% (5/40), the gene of SV40 was negative., Conclusion: Genes of BKV and JCV was detectable in urine specimens of healthy persons and there was a correlation between the load of polyomavirus and incidence of hemorrhagic cystitis.

Objective: To study the prevalence of human herpesvirus 7 (HHV-7) infection in recipients following allogeneic hematopoietic stem cell transplantation (allo-HSCT)., Methods: Peripheral blood samples were collected before and weekly after allo-HSCT from 72 consecutive recipients and 53 donors. Nested polymerase chain reaction (nPCR) was used to monitor HHV-7 DNAemia., Results: HHV-7 DNAemia was detected at least once in 86.1% (62/72) of the 72 patients on the median day 15.6 (7 approximately 56 days) after allo-HSCT. Continuing HHV-7 DNAemia (HHV-7 DNAemia maintained at least 4 weeks) was evidenced in 40.3% (29/72) of the patients after allo-HSCT. The prevalence of continuing HHV-7 DNAemia in the patients receiving HLA mismatched or HLA matched unrelated allo-HSCT who underwent conditioning with anti-thymocyte globulin (ATG) was 48.9% (23/47), significantly higher than that in the patients receiving HLA matched related allo-HSCT with conditioning without ATG [24% (6/25), P = 0.040]. Enhanced incidence the prevalence of HHV-7 DNAemia in the patients receiving corticosteroid after allo-HSCT was 44.6% (39/65), significantly higher than that in the patients who did not receive corticosteroid [0% (0/7), P = 0.037]. No relationship was observed between HHV-7 infection and transplant-related complications including acute graft versus host disease, hemorrhagic cystitis, cytomegalovirus and HHV-6 infection., Conclusion: Common in allo-HSCT recipients, HHV-7 DNAemia is closely associated with conditioning with ATG and application of corticosteroid after allo-HSCT.

This study was aimed to investigate the correlation between the serum levels of IL-18 and acute graft versus host disease (aGVHD) in patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT), and to explore the role of serum IL-18 levels in the pathogenesis of aGVHD so as to provide a reliable and early indicator for the diagnosis of aGVHD. 62 patients received allo-HSCT were enrolled in this study. Before and after transplantation, as well as at onset of aGVHD. The serum levels of IL-18 were analyzed by ELISA. 62 patients were divided into 5 groups: group A without aGVHD (28 cases) referred to the patients who had no aGVHD after transplantation and whose specimen were collected before transplantation; group B with aGVHD before transplantation (34 cases) referred to the patients who had aGVHD after transplantation and whose specimen were collected before transplantation; group C before the onset of aGVHD (34 cases) referred to patients with I - II grade a GVHD whose specimen were collected before 3 - 4 days, and according to whether the I - II grade aGVHD patients developed III - IV grade or not after treatment, these patients were divided into two subgroups retrospectively, one subgroup had good curative effect (18 cases) and the other subgroup had not (16 cases); group D with I - II grade aGVHD; group E with III - IV grade aGVHD (16 cases). The results showed that 34 patients developed I - II grade aGVHD, then out of them 16 patients (16/34) developed III - IV grade aGVHD. The serum levels of IL-18 in these patients with aGVHD were higher than that in patients without aGVHD. About 3 days before onset of aGVHD, the serum levels of IL-18 started to increase. The serum levels of IL-18 were correlated with the severity of aGVHD, but no correlation was found with infection, conditioning regimens and disparity of HLA-typing. The serum levels of IL-18 in the early stage of aGVHD were correlated with prognosis. The aGVHD of patients with higher serum levels of IL-18 easy developed to III - IV grade aGVHD. It is concluded that the serum level of IL-18 in the patients received allo-HSCT is related to the occurrence of aGVHD. Detections of serum IL-18 are helpful for the early diagnosis of aGVHD, and the serum levels of IL-18 may be considered as a reliable indicator to evaluate the prognosis and severity of aGVHD.

The study was aimed to establish a new method of preparation of human placenta factor (PF) and to determine its physic-chemical properties, as well as effects on lymphocytes in vitro. PF was prepared by ultrafiltration. The contents and molecular weight of all constitutions were determined by Bradford method and SDS-PAGE, respectively. Cyclosporin A (CsA) was served as positive control, normal saline (NS) was used as negative control. PHA-stimulated lymphocyte proliferation and mixed lymphocyte reaction (MLR) were detected with MTT assay. The expression of CD69 on T cells was analyzed by flow cytometry. Cytotoxicity of natural killer (NK) cells against K562 tumor cells was examined with LDH release assay. The results indicated that PF was determined to be a group of low molecular weight polypeptides, consisting of two major components whose molecular weight were 9.187 and 4.794 kD respectively. The contents of PF were 5.7 - 6.9 mg/g fresh placenta. PF had similar suppressive effects on PHA-stimulated lymphocyte proliferation and MLR in vitro as compared with CsA (P > 0.05). Both PF and CsA could downregulate the expression of CD69 on T cells which had been stimulated by PMA plus ionomycin (PF vs CsA, P > 0.05). The cytotoxicity of NK cells against K562 cells in PF group was slightly higher or equivalent as compared with that in NS group (P > 0.05), but the cytotoxicity in CsA group was much lower than that in NS group (P < 0.05). It is concluded that a new method of preparation of PF has been established. This study first demonstrates that PF has strong immunosuppressive effects on T cell in vitro, and suppresses T cell proliferation and activation induced by mitogen and alloantigen. This study indicats that PF has no any inhibitory effects, but even enhances the cytotoxicity of NK cells against K562 tumor cells. These results suggest that PF may have suppressive effects on graft-versus-host disease (GVHD) without diminishing graft-versus-tumor (GVT) effects. Therefore, PF may probably be an ideal and promising agent against GVHD.

The aim of study was to investigate the importance of chromosome aberration in differential diagnosis of eosinophilia and the chromosomal aberrations involved in patients with clonal eosinophilia. 65 cases of eosinophilia were collected and chromosome specimens of bone marrow cells were prepared by 24-hour culture, and G-banding technique was used for karyotyping. The results showed that out of 65 cases, chromosome 16 inversion was detected in 9 patients suspected as M(4Eo), and among the other 56 cases, 5 were detected with chromosomal aberrations (8.9%). Combining clinical, hematological and cytogenetical data, the 5 patients were diagnosed as acute myeloid leukemia with eosinophilia, chronic eosinophilic leukemia, 8p11 myeloproliferative syndrome, chronic myeloid leukemia in acute phase and acute myeloid leukemia-M(4Eo) respectively. The detected chromosomal aberrations were +14, t (5; 12) (q31; p13), t (8; 9) (p11; q32), t (9; 22) (q34; q11) and inv (16) (p13 q22). In conclusion, cytogenetical detection is very important in differential diagnosis of clonal eosinophilic disorders and chronic eosinophilic leukemia, which is suggested to be done routinely in clinic.

Objective: To study the safety and effect of G-CSF mobilized donor hematopoietic stem cells infusion (GPBSCI) as the therapy for the failure of platelet recovery after allogeneic hematopoietic stem cell transplantation., Methods: The clinical data of 15 patients with acute or chronic leukemia undergoing GPBSCI 16 times, 9 males and 6 females, aged 33 (14 approximately 48), were retrospectively analyzed., Results: The median number of mononuclear cells (MNC) from bone marrow was (4.21 +/- 1.91) x 10(8)/kg (1.50 x 10(8)/kg approximately 7.46 x 10(8)/kg). The median number of MNCs from peripheral blood was (3.27 +/- 1.40) x 10(8)/kg (1.13 x 10(8)/kg approximately 5.90 x 10(8)/kg). The median CD34+ count was (2.13 +/- 1.69) x 10(6)/kg (0.24 x 10(6)/kg approximately 5.67 x 10(6)/kg). All the patients had achieved white cell engraftment. 8 patients with primary failure of platelet recovery and 7 patients with secondary failure of platelet recovery received donor peripheral stem cells infusion. The median day of infusion was +113 days (43 approximately 384 days). The median number of infused mononuclear cells was (3.09 +/- 1.54) x 10(8)/kg (1.35 approximately 5.99) x 10(8)/kg. Only 1 patient had transfusion related GVHD. Clinical efficacy was seen in 9 patients (56.3%). The efficacy of infusion within 100 day after transplantation was 87.50%, significantly higher than that conducted 100 days after the transplantation (25.00%, P = 0.012)., Conclusion: With minimal side effect, GPBSCI may be an effective strategy for the therapy of failure of platelet recovery.

Objective: To examine cytokeratin 18(CK18) and it's gene in jaw odontogenic keratocyst (OKC) epithelial lining., Methods: The epithelial linings of 32 cases were subject to monoclonal antibody immunohistochemical staining for CK18, CK8 and CK19. RT-PCR and in situ hybridization for CK18 mRNA were conducted in 12 of 32 cases in keratocyst epithelial cell linings., Results: In 17 cases, CK18 were observed in keratinized surface layers, though weakly positive. In 27 cases, CK18 were positive in the granular cell layers. CK18 were also positive in the spinous cell layers in 14 cases. In all cases, CK18 was negative in basal cell layers. By RT-PCR, 4 cases expressed CK18 strongly, 8 cases weakly. By in situ hybridization, 8 cases expressed CK18 mRNA positively in both spinous and granular cell layers, and 4 cases positively in basal and keratinized cell layers. CK8 were expressed in basal cell layers of keratocyst epithelial linings. In 23 cases, CK19 were expressed in surface cell layers of keratocyst epithelial linings., Conclusion: The expression of CK18 in keratocyst epithelial linings transfers from basal cell layer to spinous layer. The expression of CK18 immunohistochemical staining and CK18 mRNA in situ hybridization are different, which shows CK18 might be related to proliferation of OKC epithelial linings. That suggests the existence of regulation of CK18 and CK18 mRNA expression.

Objective: To analyze the incidence and risk factors of hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation (HSCT)., Methods: The medical records of 250 patients undergoing allogeneic HSCT in Peking University Institute of Hematology from Sep. 2003 to Sep. 2005 were analyzed., Results: HC occurred in 72 of the 250 patients within 180 days after transplantation with a cumulative incidence of 28.8% (SE 0.3%). None of early-onset HC was developed in the cohort. The median time of onset was 33 days after HSCT (range 14 - 170 days) and the median duration of HC was 35 days (range 3 -186 days). There were HC of grade I - II in 51/72 cases (70.83%) and of grade III - IV in 21/72 (29.17%). Univariate analysis indicated that age younger than 25, high risk disease, CMV reactivation, ATG usage, graft from MUD or MMRD and GVHD grade IL - IV were associated with the occurrence of HC, while in multiple regression analysis only GVHD grade II - IV (RR = 2.75; 95% CI 1.63 -4.66; P < 0.01) and donor type (RR = 2.60; 95% CL 1.52 - 5.20; P < 0.01) were independent risk factors. Kaplan-Meier survival analysis indicated HC not increasing the mortality (RR = 0.67, 95% CI 0.33 - 1.36)., Conclusion: HC post allo-HSCT is a common complication, GVHD grade II - IV and donor type are the independent risk factors.

Background: Allogeneic stem cell transplantation is a potential curative approach in patients with multiple myeloma. The very high transplant related mortality associated with standard allogeneic stem cell transplantation is currently the major limitation to wider use of this potentially curative treatment modality. The challenge for clinical investigators is to reduce the incidence of post-transplant complications for patients receiving autologous hematopoietic stem cell transplantion for multiple myeloma. In this study the toxicity and efficacy of modified myeloablative conditioning regimen followed by allogeneic stem cell transplantation was investigated in patients with multiple myeloma., Methods: The conditioning regimen consisted of hydroxyurea, cytarabine, busulfan, cyclophosphamide, and semustine. Ten patients underwent allogeneic transplantation among them hydroxyurea (40 mg/kg) was administered twice on day-10 and cytarabine (2 g/m(2)) was given on day-9, busulfan was administered orally in four divided doses daily for 3 days (days-8 to -6). The dose of busulfan was 12 mg/kg in the protocol followed by cyclophosphamide intravenously over 1 hour on days-5 and -4 (1.8 g/m(2)), and with semustine (Me-CCNU) 250 mg/m(2) on day -3., Results: Chimerism data were available on all patients and all patients achieved full donor chimerism without graft failure. Six patients had not acute graft-versus-host disease (GVHD, 36.4%; 95% CI: 13.9% - 38.6%). Two patients (18.2%) developed grade I acute GVHD (95% CI: 10.9% - 35.9%) and grade II acute GVHD occurred in one patient (9.1%; 95% CI: 8.4% - 32.3%). Severe grade IVa GVHD was seen in one patient, who died from acute GVHD. The incidence of chronic GVHD was 22.2% (95% CI: 11.7% - 36.7%), among them one died of severe grade IV GVHD and one developed multiorgan failure on day +170; the treatment-related mortality was 22.0% (95% CI: 10.3% - 34.1%). The overall 4-year survival rate was 67.8% (95% CI: 16.3% - 46.7%). The estimated 4-year progression-free survival rate was 58.5% (95% CI: 13.7% - 41.8%). The 4-year complete remission was 72.7% (95% CI: 27.8% - 49.6%). One patient relapsed after 4 months and achived the complete remission after receiving the donor lymphocyte infusion., Conclusions: Modified conditioning regimen busulfan-cyclophosphamide with peripheral blood stem cells + bone marrow cells transplantation result in a low incidence of severe GVHD with a relatively low treatment-related mortality, high complete remission rates and a long-term survival.

The aim of study was to investigate the pharmacokinetics and distribution of antithymocyte globulin (ATG) in recipients of partially HLA-matched hematopoietic stem cell transplantation. Fifteen patients with hematological disorders were received hematopoietic stem cell transplantation from partially HLA-matched related donor between October 2003 and October 2004 in the Institute of Hematology and People Hospital, Peking University. All patients including 5 cases of AML, 6 cases of CML, 3 cases of ALL, 1 case of AA were consecutively enrolled in the present study after providing written informed consent. Antithymocyte globulin was administered before allogeneic hematopoietic stem cell transplantation at a dose of 2.5 mg/kg daily for 4 consecutive days (total dose of 10 mg/kg) in the conditioning regimen. The concentration of rabbit ATG in the serum of 15 patients was measured using a new enzyme-linked immunoabsorbent assay (ELISA) for the Fc portion of rabbit IgG. The results showed that the washout phase of ATG elimination was analyzed over 0 - 120 days, results were well-fitted by a single exponential decay giving a mean elimination half-life (t(1/2) beta) of 29.67 +/- 2.60 days. A mean value for the apparent volume of distribution of ATG (V) obtained by analysis of data was 0.12 +/- 0.02 L/kg body weight. The serum concentration of ATG increased up to 44.8% at 5 day before transplantation, peak concentration of ATG was 136.0 +/- 10.3 mg/L, its concentration slowly descend at 0 day, fall up to 7.1 +/- 0.06 microg/ml at 90 day after dosing; t(max) 4.8 +/- 0.7 days; According to AIC (Akaike's information criterion), two compartment model of ATG was estimated. It is concluded that the conditioning regimen containing the dosage of 10 mg/kg of ATG is effective and safely in recipients of partially HLA-matched hematopoietic stem cell transplantation. There is no racial difference in the pharmacokinetics of ATG.

To investigate the cytogenetic characteristics of multiple myeloma and its relationship with clinical prognosis, 68 cases were collected and chromosome specimens of bone marrow cells were prepared by 24-hour culture, and G-banding technique was used for karyotype analysis. The results showed that the detected chromosome aberration rate was 19.1% (13/68). The abnormal clones existed mosaically with normal clones. Numerous aberrations were manifested by aneuploidy, mainly by hyperdiploidy or hypodiploidy. Structural aberrations involved t (11; 14), chromosome 1 and various kinds of marker chromosomes. Cases which had very complex aberrations revealed poor prognosis. It is concluded that chromosome complex aberration is the mainly cytogenetic characteristics of multiple myeloma, and multiple numerous and structural aberrations are involved. Cytogenetic detection should be performed both at diagnosis and at disease progression so as to evaluate prognosis.

In order to explore the application of real-time quantitative reverse-transcription polymerase chain reaction (Q-PCR) for detecting PML/RARalpha gene transcripts in patients with acute promyelocytic leukemia (APL), the bone marrow samples from 46 newly diagnosed APL patients were collected for analysis. Three plasmids containing cDNA fragments of the bcr1-, bcr3-form PML/RARalpha and ABL control gene were constructed respectively. The ABI Prism 7500 Sequence Detection System using Taqman fluorogenic probes was used to quantify target gene. PML/RARalpha mRNA was detected by Q-PCR in 46 APL patients and 40 non-APL patients. The normalized quotient (NQ) of PML/RARalpha mRNA was calculated as followings: NQ = PML/RARalpha mRNA copy numbers/ABL mRNA copy numbers. Immunophenotype of acute promyelocytic leukemia was determined by four-color flow cytometry. The results showed that the coefficients of variation (CV) of inter-day assay and intra-day assay by using Q-PCR were 1.58% and 0.88% respectively. Q-PCR could detect reproducibly 5 copies of target gene per 100 ng RNA and no pseudopositive results were found. The median NQ of PML/RARalpha mRNA was 0.450 (0.084 - 1.082) in 46 APL patients. There was no indication of any correlation of PML/RARalpha mRNA type with age, sex, hemoglobin, platelet count, percentage of promyelocytes in bone marrow detected by morphology or flow cytometry, PML/RARalpha NQ, or signs of clinically diagnosed coagulation/bleeding disorders. Compared with bcr1-form cases, bcr3-form cases had more M(3v) phenotype (42.9% vs 9.4%, P = 0.015) and higher WBC count (9.35 x 10(9)/L vs 2.15 x 10(9)/L, P = 0.038). APL cells could be classified into large side scatter population (L-SSC) and non-large side scatter population (NL-SSC) in CD45/SSC histogram of flow cytometry. 87.50% patients with bcr1-form showed L-SSC phenotype and 64.29% patients with bcr3-form showed NL-SSC phenotype. It is concluded that a sensitive Q-PCR method is established. The median NQ of PML/RARalpha mRNA was 0.450 in newly diagnosed APL patients. There was no significant difference about PML/RARalpha mRNA expression of both bcr3-form and bcr1-form APL patients. Type of PML/RARalpha transcripts is related with the morphology and immunophenotype.

Objective: To analyze the etiology and clinical features of late onset hemorrhagic cystitis (LOHC) after allogeneic hematopoietic stem cell transplantation (HSCT)., Methods: The medical records of 200 patients undergoing allogeneic HSCT from 2004 to 2005 were analyzed retrospectively., Results: HC developed in 57 patients within 180 days after the transplantation with a cumulative incidence of 28.8%. The etiology of 31 patients (54.39%) was infection, caused by infection of cytomegalovirus (CMV) or adenovirus and cured by anti-virus therapy, thus the cause of disease could be classified as infection agent. Viremia was seen in 12 patients (21.53%) with CMV and disappeared in urine after anti-virus therapy but bleeding still persisted. For these patients the cause of disease was classified as infection agent combined with non-infection factor. Evidence of infection agent could not be discovered in 14 patients (24.56%) and they failed to respond to anti-infection therapy. For them the cause of disease was classified as non-infection agent. 13 patients with refractory HC underwent tentative treatment with corticosteroids, 9 of them achieved a complete remission, 2 of them achieved partial remission, and 2 of them remained non-responsive., Conclusion: LOHC after allo-HSCT is a common complication and caused by multiple factors. Differentiation of the possible causes may benefit its clinical outcome.

Using Bi-Digital O-Ring Test Resonance Phenomena between 2 identical substances, Omura, Y. succeeded in making the image of the outline of internal organs without use of standard imaging devices since 1982. When he imaged the outline of the stomach on the abdominal wall, a number of the lines came out from upper and lower parts of stomach wall. When the lines were followed, they were very close to the well-known stomach meridians. Subsequently, he found a method of localizing meridians and their corresponding acupuncture points as well as shapes and diameters accurately. At the anatomical location of ST 36 described in traditional textbooks, Omura, Y. found there is no acupuncture point. However, in the close vicinity, there is an acupuncture point which he named as true ST 36 in the mid 1980s, but it is generally known as Omura's ST 36. When the effects of the acupuncture on these 2 locations were compared, Omura's ST 36 (true ST 36) produced very significant well-known acupuncture beneficial effects including improved circulation and blood chemistry, while in the traditional ST 36, the effects were small. In this article, the anatomical relationship between these two acupuncture points, with a short distance of 0.6 approximately 1.5 cm between the centers of these locations, was described. In early 2000, Omura, Y. found Press Needle Stimulation of Omura's ST 36, using "Press-Release" procedure repeated 200 times, 4 times a day to cancer patients reduced high cancer cell telomere of 600-1500ng and high Oncogen C-fos Ab2 and Integrin alpha5beta1 of 100-700ng BDORT units to close to lyg (= 10(-24) g) BDORT units. In addition there was a significant reduction of Asbestos and Hg from cancer cells, while markedly reduced normal cell telomere of lyg was increased to optimally high amounts of 500-530ng BDORTunits. Thus, cancer cells can no longer divide and cancer activity is inhibited. The authors have successfully applied this method for a variety of cancers as well as for cardio-vascular diseases with hypertriglyceridemia, hyperglycemia, high L-homocystein, and CRP, high cardiac Troponim I & T, and some hypertension. These beneficial effects were accompanied by euphoria, & relaxation with increased alpha waves in EEG. Thus Omura's ST 36 stimulation is a safe, effective and highly desirable supplemental treatment. In addition to manual stimulation, similar beneficial effects can be induced by finger tip stmulation (without any needle) or with electroacupuncture stimulation, (+) Qi Gong energy stored paper and (+) solar energy stored paper which often resulted in significant clinical improvement.

To study feasibility of HLA-DRB1 matching by using denatured high performance liquid chromatography (DHPLC), 20 pairs of DNA samples from donors and recipients of hematopoietic cell transplantation (HCT) for DRB1 matching and 2 pairs of samples from donors and recipient of HCT for DRB1 mismatching were studied by DHPLC and PCR-SSP. After being amplified and annealed slowly to produce heteroduplex, PCR products for exon 2 of DRB1 were detected by DHPLC to find matched or mismatched peaks in chromatogram. The results showed that DHPLC and PCR-SSP were consistant with matched or mismatched HLA-DRB1 typing. The results of DHPLC and PCR-SSP for matching were compared by using kappa test (kappa = 0.776, P = 0.00), which suggested DHPLC for HLA-DRB1 matching was in agreement with PCR-SSP. In conclusion, DHPLC for HLA-DRB1 matching is economic and convenient, moreover, will not be affected by unknown genes in HLA-DRB1 locus.

In order to study the prevalence of human herpesvirus 6 (HHV-6) in allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients in China and to analyze the relationship between HHV-6 and cytomegalovirus (CMV) infection in post-HSCT patients, nested polymerase chain reaction (PCR) was used to monitor HHV-6 DNAemia in 72 consecutive allo-HSCT recipients. 680 EDTA anticoagulated peripheral blood specimens were gathered before HSCT or weekly until 12 weeks after allo-HSCT. The variants of HHV-6 were identified by Hind III restriction analysis. CMV-pp65 antigenemia was detected by immunofluorescence stain. The results showed that HHV-6 DNAemia was detected at least once in 62.5% (45/72) of the patients on the median day 14 (range, 7 - 63 days) after allo-HSCT, and HHV-6B was the predominant variant. CMV antigenemia was detected at least once in 65.3% (47/72) of the patients on the median day 43 (range, 14 - 105 days) after allo-HSCT. Co-infection of HHV-6 and CMV (HHV-6+/CMV+) occurred in 52.8% (38/72) recipients. The onset of HHV-6 DNAemia was earlier than that of CMV antigenemia (P < 0.0001). Patients with HHV-6 DNAemia positive were more likely to have concurrent CMV antigenemia than HHV-6 DNAemia negative patients (84.4% vs 33.3%, P = 0.0001) after allo-HSCT. Among the herpesvirus related disease, the relatively high incidence of hemorrhage cystitis (HC) occurred in 23.6% (17/72) of post-HSCT patients. 88.2% (15/17) of HC developed in HHV-6 positive patients, and 82.3% (14/17) occurred in CMV+/HHV-6+ patients. It is concluded that infection of HHV-6, co-infection of HHV-6 and CMV, commonly occurred in post-HSCT patients in China, HHV-6 infection closely related to CMV antigenemia.

Objective: To investigate the role of killer cell immunoglobulin receptor (KIR) plays in treatment of leukemia by cord blood transplantation (CBT)., Methods: Peripheral blood samples were collected from 23 patients receiving CBT, and corresponding cord blood samples were collected too. Polymerase chain reaction with sequence-specific primers was used to examine the genotypes of KIR and HLA-Cw of the samples. The rates of engraftment, relapse, and survival were followed up., Results: Polymorphism of KIR genotype was found in the 23 cord blood samples. In the 17 cases of acute myelocytic leukemia (AML), the engraftment rate and overall survival rate of the donors/recipients with KIR/JHLA-C mismatch was 65% and 70%, both higher than those of the KIR/HLA-C matched group (50% and 50% respectively), and the relapse rate of those with KIR/JHLA-C mismatch was 9%, lower than that of the KIR/HLA-C matched group (16%). The engraftment rate and overall survival rate of those with homo-expression of HLA-Cw1 or Cw2 were 65% and 70% respectively, both higher than those of the patients with double expression (50% and 50% respectively). There was no significant difference in overall survival rate between the HLA-Cw8 (+) and HLA-Cw8 (-) groups. Four cases with the co-expression of KIR2DL2 and KIR2DS2 all survived disease-free., Conclusion: KIR plays an important role in CBT. KIR/HLA-C mismatch, homo-expression of HLA-Cw1 or Cw2, and co-expression of KIR2DL2 and KIR2DS2 may predict good prognosis for CBT.

We describe the results of a clinical trial to evaluate the feasibility and toxicity of autologous hematopoietic stem cell transplantation (auto-HSCT) for patients with progressive multiple sclerosis (MS). Fifteen patients (all patients with secondary progressive MS) were enrolled. The median expanded disability status scale (EDSS) score at baseline was 6.0 (range, 4.5-7.5). Peripheral blood stem cells were obtained by leukapheresis after mobilization with granulocyte colony-stimulating factor. In 9 patients, CD34+ cell selection was performed with a CliniMACS cell selection system, and 6 patients accepted infusion of unmodified peripheral blood stem cells. The modified BEAM (carmustine, teniposide, cytarabine, and melphalan) was the sole conditioning regimen used. The adverse effects included infections, mucositis, transient hepatotoxicity, and diarrhea. Three patients had flares of neurologic deterioration during mobilization, 8 patients had the same manifestation during transplantation, and 2 patients had similar flares within 3 months of transplantation. Six patients experienced continuous neurologic improvement after transplantation, 5 patients experienced neurologic progression, and 4 patients had stabilization of their disease. The confirmed progression-free rate was 63.8% at 49 months. The results of lymphocyte purging were no better than for no purging. Auto-HSCT proved to be safe and beneficial for some MS patients. Further studies are needed to establish the merit of this procedure for MS patients.

To evaluate the significance of FCM in minimal residual disease (MRD) detection, the immunophenotyping and leukemia-associated immunophenotypes (LAIP) of leukemia cells from 273 adult and 142 childhood patients with B lineage acute lymphoblastic leukemia (B-ALL) were detected by four to six antibody combinations of 4-color CD45/SSC gating multiparametric flow cytometry (FCM). The results showed that the B-ALL patients could be classified into 4 subtypes based on different expression CD34 and CD10: subtype I (CD34(+)/CD10(-)), subtype II (CD34(+)/CD10(+)), subtype III (CD34(-)/CD10(+)), subtype IV (CD34(-)/CD10(-)). The LAIP was observed in 100% and 92% patients of subtype I and subtype II, respectively, whereas only 79.2% in subtype III. The incidence of LAIP in total B-ALL cases was 90% by using the antibodies detected in this investigation. There was no significantce different for incidence of LAIP between adult and pediatric patients. LAIP was observed in 77.6% of patients by labeling only CD34/CD10/CD19/CD45 4-color antibody combination. It is concluded that in 90% of childhood and adult B-ALL patients LAIP can be found, which suits MRD detection by multiparameter flow cytometry.

Although human herpesvirus 6 (HHV-6) has been considered an important opportunistic and potentially fatal pathogen for allogeneic hematopoietic stem cell transplantation (HSCT), the clinical significance of HHV-6 reactivation remains controversial. In this study, we monitored HHV-6 DNAemia in 72 consecutive allogeneic HSCT recipients by real-time quantitative polymerase chain reaction. A total of 680 peripheral blood specimens were collected from the recipients before HSCT or at weekly intervals after HSCT. As the predominant variant, HHV-6B was detectable at least once in 47.2% (34/72) of HSCT recipients on the median day 21 (range, 7-84 days); HHV-6A reactivation occurred in only 1 recipient (1.4%). Detectable HHV-6B reactivation was associated with increased probability of skin rash by day 30 after HSCT (hazard ratio [HR], 3.68; 95% confidence interval [CI], 1.24-10.92; P = .019), cytomegalovirus (CMV) antigenemia (HR, 2.35; 95%CI, 1.32-4.19; P = .004), and hemorrhagic cystitis (HC) (HR, 2.59; 95%CI, 0.96-6.98; P = .061) by day 100 after HSCT. Neutrophil and platelet engraftment, mortality for 100 days after HSCT were not affected by HHV-6B reactivation. In conclusion, HHV-6 reactivation is a common event, and this study demonstrates a correlation between HHV-6B infection and CMV reactivation, early rash, and possibly increased incidence of HC after transplantation.

Human herpesvirus 6 (HHV-6) reactivation was studied in 72 consecutive allogeneic hematopoietic stem cell transplant (HSCT) recipients and 53 "healthy" donors. The feasibilities of real-time quantitative polymerase chain reaction (RQ-PCR), nested-PCR, and antigenemia assays in the assessment of HHV-6 reactivation were also evaluated. HHV-6 DNA was detected in 62.5% and 48.6% of post-HSCT patients with the nested-PCR assay and RQ-PCR analysis, respectively, and HHV-6B was identified as the predominant variant. The incidence of HHV-6 infection peaked from the second to the seventh week, whereas the HHV-6B DNA loads peaked from the second to the third week. Compared with RQ-PCR analysis, the sensitivity and specificity of the nested-PCR assay were 100% and 88%, respectively, with positive and negative predictive values of 60% and 99%, respectively. For the HHV-6 antigenemia assay, the sensitivity and specificity were 89% and 97%, respectively, and the positive and negative predictive values were both 94%. Conditioning with antithymocyte globulin in HLA-mismatched or unrelated HSCT increased the possibility of HHV-6B reactivation after HSCT (hazard ratio, 5.92; 95% confidence interval, 1.99-17.59; P = .001). In conclusion, HHV-6B reactivation is commonly encountered after HSCT. Of the 3 methods we adopted for HHV-6 detection, both RQ-PCR analysis and the antigenemia assay could be seen as essential tests for predicting HHV-6 reactivation.

This article presents a practical approach for safe oral sedation in the dental practice. When used properly, oral sedation can provide comfort and a calming treatment environment for patients whose fear inhibits them from securing needed dental care. In Part I, the authors provided information on medico-legal aspects of sedation, patient treatment recommendations, counseling, evaluation, monitoring, documentation, and proper discharge procedures. In this part, the reliable sedatives that have undergone years of clinical trials and have good records of safety and predictable results are presented. For each sedative, the description, formulation and dosage, onset, duration of sedation, and side effects are discussed.

Objective: To explore the prognosis and cytogenetic characteristics of acute erythroid leukemia., Methods: We selected 55 patients with acute erythroid leukemia and reviewed the cytogenetic characteristics of these cases. With case-control studies, 55 patients were classified as primary or MDS transformed leukemia depending on the absence or presence of MDS history. In addition, we also classified these 55 cases as normal or abnormal karyotype. We analyzed the treatment response to allogenic hematopoietic stem cell transplantation or chemotherapy and the survival and prognosis of all the groups with Kaplan-Meier and Log-Rank., Results: 45 patients and cytogenetic studies and there were 18 cases with normal karyotypes and 27 cases with aberrant karyotypes. The detection rate of chromosomal abnormality was 60.0%. In the cases with chromosomal abnormality, there were 17 cases with multiple chromosomal abnormality and 10 cases with simple chromosomal abnormality. 10 cases had excess of hypodiploid or hyperdiploid. In 18.5% (5/27) of the cases with aberrant karyotypes 5 chromosomes were involved and in 25.9% (7/27) 7 or 8 chromosomes were involved. The 55 patients were followed up and their treatment response was evaluated. In these patients, total complete remission rate was 63.6%. Among them, the complete remission rate of acute erythroid leukemia with MDS history was lower than that without MDS history (42.8% vs 85.2%, P < 0.05). The complete remission rate of acute erythroid leukemia with aberrant karyotypes was lower than that with normal karyotypes (37.0% vs 83.3%, P < 0.01). The median follow-up was 30 months (3 - 79 months). During follow-up, the overall survival and disease-free survival of the patients with aberrant karyotypes or MDS history who received transplantation were longer than those who received chemotherapy (P < 0.01). Chromosomal abnormality and previous MDS history were the major factors that influenced prognosis. 16 patients received allogeneic haemopoietic stem cell transplantation. Among them, 9 cases had aberrant karyotypes and previous MDS history and 4 cases had no response to chemotherapy. After transplantation, 11 cases survived without any disease. The median survival was 28 months and 2-year survival 68.7% (11/16)., Conclusions: Acute erythroid leukemia patients with aberrant karyotypes or MDS history are usually resistant to chemotherapy. The survival of these patients is short and the prognosis poor. Allogeneic haemopoietic stem cell transplantation could improve survival and prognosis. The patients with aberrant karyotypes or MDS history should receive allogeneic haemopoietic stem cell transplantation as early as possible.

This article presents a practical approach for safe oral sedation in dentistry. When properly done, oral sedation can provide comfort and a calming treatment environment for patients whose fears inhibit them from securing needed dental care. The authors provide information on medico-legal aspects of sedation, patient treatment recommendations, counseling, evaluation, monitoring, documentation, and proper discharge procedures. The reliable sedatives that have undergone years of clinical trials with a good safety record and predictable results are presented. For each sedative drug, the description, formulation and dosage, onset, duration of sedation, Oral and side effects are discussed in a 2-part presentation.

Objective: To study the potential relationship between HHV-6 activation and acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic cell transplantation (HSCT)., Methods: Peripheral blood samples were collected before and weekly after HSCT from 72 consecutive recipients. HHV-6 DNAemia was monitored by nested polymerase chain reaction (PCR). The genotypes of HHV-6 were identified by Hind III restriction assay., Results: Of the 72 patients, HHV-6 DNAemia were detected in 45 (62.5%) on a median of day 14 (range, 7 - 63 days) after HSCT. Grade I - IV aGHVD occurred in 40 (55.6%) on a median of day 26 (range, 9 -73 days). The median onset time of HHV-6 DNAemia was significantly earlier than that of aGHVD (P = 0.018). Compared with that in HHV-6 DNAemia negative [HHV-6(-)] patients, the cumulative incidence of grade I - IV aGHVD was higher (68.9% vs. 33.3% , P = 0.003) in HHV-6 (+) patients. Cumulative incidence of grade II - IV aGVHD in HHV-6 (+) cohort was also higher than that in HHV-6 (-) cohort (35.6% vs 14.8% , P = 0.027). Cumulative incidence of grade I - IV aGVHD was higher in patients with both HHV-6 and CMV positive (CMV+/HHV-6+) than in those with either CMV (CMV+/HHV-6-) or HHV-6 positive (CMV+/HHV-6+) and neither of them positive (CMV-/HHV-6-) [78.9% (30/38), 55. 6% (5/9) , 14. 3% (1/7) and 22. 2% (4/18), respectively, P = 0. 0001]. Cumulative incidence of grade II - IV aGVHD in CMV+/HHV-6+ group was also higher than that in CMV+/HHV-6-, CMV-/HHV-6+ and CMV-/HHV-6- groups [42.1% (16/38), 22.2% (2/9), 0% (0/7) and 11.1% (2/18), P = 0. 008]., Conclusions: Patients with HHV-6 activation or HHV-6/CMV co-infection maybe involved in the occurrence of aGVHD after HSCT.

Objective: To explore the indication and optimum time for treating myelodysplastic syndrome (MDS) by allogeneic hematopoietic stem cell transplantation (allo-HSCT) with HLA identical sibling grafts., Methods: From June 1997 to Sep. 2004, a total of 30 patients with MDS were treated with allo-HSCT from HLA-identical sibling donors in our institute. On HSCT, 4 patients had refractory anemia (RA) , 2 RA with ringed sideroblasts (RARS) , 7 RA with excess blasts(RAEB) , 14 RAEB in transformation (RAEB-t) , 3 already progressed to secondary AML. For IPSS system, 6 patients were in intermediate- I risk group, 11 in intermediate- li risk group, and 13 in high risk group. The modified BU/CY conditioning regimen was used. Four patients received bone marrow transplantation (BMT), 8 received peripheral blood stem cell transplantation (PBSCT) , and 18 received BMT + PBSCT., Results: The 3-year expected overall survival (OS) was 63.61%, 3-year expected disease-free survival ( DFS) 61.41%, and relapse rate 5.26%; OS for RA/ RAS, RAEB and RAEB-t/AML subgroup was 83.33%, 34.29% and 66.67% , respectively, and all had no statistic difference among them. OS for IPSS-intermediate and high risk subgroup was 64.7% , and 69.0% respectively, also had no statistic difference. 3-year expected OS in no aGVHD,grade I - II aGVHD and grade III - IV aGVHD group was 57.75% , 100% and 0% , respectively (P = 0.009). Pre-HSCT chemotherapy, disease subtype and cGVHD all had no correlation with LFS or OS (P > 0.05)., Conclusion: For young MDS patients having HLA-identical sibling donors, HSCT should be the first line therapy and performed as soon as possible.

Many patients who require allogeneic hematopoietic stem cell transplantation (allo-HSCT) lack a human leukocyte antigen (HLA)-matched donor. Here, we report a protocol for haploidentical allo-HSCT that combines granulocyte-colony stimulating factor primed bone marrow (G-BM) and peripheral blood stem cells (PBSC) without in vitro T-cell depletion (TCD). In this study, 171 patients, including 86 in high-risk group, underwent transplantation from haploidentical family donors. All patients achieved sustained, full donor chimerism. One hundred and eleven patients were alive in remission at a median of 682 (253-1502) days. The cumulative incidence of grade III-IV acute graft-versus-host disease (GVHD) was 23% and that of extensive chronic GVHD, 47%; these were not influenced by HLA disparity. Patients younger than 15 years had less grade III-IV acute GVHD than older patients (P=0.044). The 2-year probability of relapse was 12% for standard-risk disease and 39% for high-risk disease. The 2-year probability of leukemia-free survival (LFS) was 68% for standard-risk patients and 42% for high-risk patients (P=0.0009). Grade III-IV acute GVHD was associated with better LFS (P=0.0017). The results require confirmation and show that G-BM combined with PBSC from haploidentical family donors, without in vitro TCD, may be used as a good source of stem cells for allo-HSCT.

The study was aimed to investigate the pp65 antigen of human cytomegalovirus (CMV) and its clinical significance in patients revived allogeneic hematopoietic stem cell transplantation (HSCT). 104 patients received allogeneic HSCT were studied. Anticoagulant blood samples were obtained from the recipients before and after transplantation and in the convalescence. CMV pp65 antigen in leukocytes was detected by indirect immunofluorescence assay using CMV Brite Kit weekly. The results showed that among the 104 patients, 29 cases were CMV pp65 positive (27.88%). Out of 29 cases 16 were CMV antigenemia and 13 cases were CMV disease. There were 25 cases who positively responded to antiviral therapy (effective ratio 86.21%) and 4 cases died (case-fatality ratio 13.79%). The detection revealed a significant difference in the incidence of CMV infection between the patients received unrelated or haploidentical family donor HSCT (39.29%) and HLA-identical sibling donor HSCT (14.58%) (P < 0.05). The incidence rate of CMV infection in patients with 0-I grade aGVHD and patients with II-IV grade aGVHD were 19.44% and 46.88% respectively, which had significant difference (P < 0.05). There was significant difference in the occurrence of aGVHD between the patients with and without positive CMV pp65 (P < 0.05). It is concluded that infection of CMV can be detected by the CMV pp65 monoclonal fluorescence immunohistochemistry, The detection of CMV pp65 antigen in peripheral blood leukocytes as a indicator for CMV disease surveillance after HSCT, which may be used to early diagnose the CMV infection, to guide the antiviral treatment and evaluate its efficacy.

Objective: To evaluate the value of real time quantitative RT-PCR (Q-PCR) for monitoring bcr-abl mRNA levels in chronic myeloid leukemia (CML) patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT)., Methods: Quantification of bcr-abl mRNA was performed on 316 bone marrow samples from 112 patients with CML after HSCT by Q-PCR using the TaqMan probe system. The bcr-abl mRNA level was normalized by control gene abl. Cytogenetic response was evaluated with fluorescent in-situ hybridization (FISH)., Results: The reproducible sensitivity of Q-PCR was 5 copies. The coefficients C(T) of interassay and intraassay variation for abl and bcr-abl were all below 2.0%. 289 bone marrow samples were collected from 101 CML patients who achieved a sustained complete cytogenetic response (CCyR) one month post allo-HSCT in a period of 6 - 60 months (median 12 months) at different intervals. In general, the median bcr-abl levels gradually decreased with the prolongation of time after HSCT: the median bcr-abl levels were 0.035% (0 - 0.406%) at 1 month post allo-HSCT (+ 1 month), 0.006% (0 - 0.683%) at +3 month, 0% (0 - 0.225%) at +6 month and remained 0% till +24 months. The highest level in CCyR patients detected at + 6 month was 0.068%. The bcr-abl mRNA level was decreased by 3 log in sustained CCyR patients at + 1 month compared with the newly diagnosed CML-CP patients (33.0%, data unpublished). On the contrary, Q-PCR results ranged from 0.12% to 13.45% in 8 cytogenetic non-responders or relapsed patients post allo-HSCT. Among them, 5 patients' samples were collected 1 - 2 months before cytogenetic relapse, the results were ranged from 0.09% to 3.42%. If 0.09% was assumed 0.09% as a threshold, 9 sustained CCyR patients (8.9%) were tested once higher than that within 6 month after HSCT but decreased to 0% eventually. 2 blast crisis patients achieved CCyR within 1.6 and 3 months after HSCT, but hematological relapse occurred after 1 and 1.5 months, and their bcr-abl mRNA levels increased dramatically from 0% and 0.14% to 46.9% and 75.9% respectively., Conclusions: Q- PCR is a sensitive, precise and reliable technique, and can be used to monitor CML patients post allo-HSCT regularly. Patients in blast phase of CML should be monitored more frequently.

Objective: To compare the outcomes in hematological patients receiving unrelated peripheral blood stem cell transplants (UPBSCT) with those receiving unrelated bone marrow transplants (UBMT) in a retrospective analysis., Methods: Sixty-three patients with hematological diseases with HLA-matched (65%) or mismatched (35%) unrelated donors were receiving UPBSCT (n = 33) or UBMT (n = 30) between May 2001 and June 2005 in our institute. They all received standard conditioning regimens with Bu/Cy (n = 53) or TBI/Cy (n = 10) with the addition of ATG for 3 - 4 days (n = 58) or CD3 antibody (n = 5). There were more patients receiving rabbit ATG in the UPBSCT group than UBMT (21/33 vs 8/25, P = 0.02) and the remaining patients received pig ATG. Graft versus host disease (GVHD) prophylaxis consisting of cyclosporine, methotrexate and mycophenolate mofetil were the same., Results: The two groups were matched for the following factors: gender, diagnosis, HLA-compatibility and conditioning regimens. The median age in the UPBSCT group was 29.5 (10 - 47) years and in the UBMT 21.5 (7 - 42) years (P < 0.05). The UPBSCT group consisted of 10 females and 23 males and the UBMT group 7 females and 23 males. Eleven patients in the UPBSCT and 8 patients in the UBMT group were diagnosed with as chronic myeloid leukemia (CML); 9 and 12 as acute myelocytic leukemia (AML); 11 and 10 as acute lymphatic leukemia (ALL); 2 and 0 as severe aplastic anemia (SAA). There were more patients in aggressive stage (>CR1) in the UBMT group than in the UPBSCT group (9/30 vs 3/33, P = 0.06). Median follow-up was 12 months after UPBSCT and 20 months after UBMT (P < 0.05). UPBSCT group had a higher number of infused MNC as comparison with UBMT group (6.16 x 10(8)/kg vs 2.80 x 10(8)/kg, P < 0.05). Both neutrophil and platelet recovery were faster after UPBSCT (11 days vs 17 days, 14 days vs 27.5 days, P < 0.01). Although the cumulative incidence of grades II - IV acute GVHD and severe aGVHD in the UPBSCT group were less than in the UBMT group (33.04% vs 66.69%, 7.26% vs 34.52%, P < 0.05), there was no difference after the source of ATG was counted. The incidence of chronic GVHD did not differ between the two groups (18/25 vs 14/22). Relapse including molecular relapse occurred in 6 of the 31 patients after UPBSCT and in 4 of the 30 patients after UBMT (P > 0.05). Finally, fifteen of the 30 patients died after UBMT, as compared with 8 of the 33 patients after UPBSCT. The cumulative overall survival was 62.45% after UPBSCT and 47.22% after UBMT (P = 0.114)., Conclusion: Our results indicate that UPBSCT led to significantly faster leukocyte and platelet engraftment without increasing the incidence of aGVHD and the overall survival was comparable between the two methods of therapy.

Objective: To explore the specificity of anti-phosphotyrosine monoclonal antibody PY20 in bcr-abl+ cells and its possible clinical applications., Methods: Bcr-abl cell lines( K562, MEG-01) and bcr-abl- cells lines( Jurkat, MCF-7 )were stained with PY20. Phosphotyrosine protein of K562 and MEG-01 cells was detected by flow cytometry before and after treatment with imatinib. Phosphotyrosine protein in bone marrow cells from 49 patients with chronic myeloid leukemia (CML), Ph+ acute lymphoblastic leukemia(Ph(+) -ALL), Ph- ALL, acute myeloid leukemia (AML-M1, M2, M3, M5, FAB classification), chronic lymphocytic leukemia (CML) and 3 normal donor. Positive cells over 5% of total cells was considered positive cases for phosphotyrosine protein. The level of tyrosine phosphorylation was determined by median fluorescence intensity (MFI)., Results: Bcr-abl cell lines and marrow cells from 10 CML patients and 8 ALL patients were all PY20-positive, while bcr-abl- cell lines and marrow cells from 18 leukemia patients and 3 normal donor were all PY20-negative. MFI decreased remarkably after blocked by imatinib in K562 cells and MEG-01 cells. The positive cell percent of marrow cells from 10 newly diagnosed CML patients and 9 imatinib-sensitive CML patients was (54.20 +/- 19.82)% and (14.84 +/- 6.17)% (P < 0.05), while that of 2 cases of imatinib-resistant was 64.3% and 57.2%. There was significant difference of MFI between imatinib-resistant patients and imatinib-sensitive patients (99.42 +/- 4.87 vs 46.41 +/- 4.67, P < 0.01)., Conclusion: PY20 monoclonal antibody is highly specific for bcr-abl+ cells. It might be useful in rapid detection of bcr-abl+ cells and sensitivity to imatinib of CML patients.

Objective: To investigate the mechanism of tetra-arsenic tetra-sulfide (As4S4) in inducing apoptosis of acute promyelocytic leukemia (APL) cells., Methods: The gene expression patterns in NB4 cells pre- and post-treatment with As4S4 were analyzed by cDNA microarray, and differentially expressed genes related with apoptosis were identified. The mRNA expression levels of these apoptosis related genes in the peripheral blood of APL patients treated with As4S4 pre- and post-remission were examined by RT-PCR., Results: Among the differentially expressed genes in NB4 cells pre- and post-treatment with As4S4, two genes were related with cellular apoptosis, which were Apaf-1 and PNAS-2. Apaf1 ratio between pre- and post- As4S4 in NB4 cells was 2.910, PNAS-2 ratio was 0.420. RT-PCR results showed that the expression ratios of Apaf-1, caspase-9 and PNAS-2 in APL patients pre- and post-remission were 2.31 and 3.21, 0.99 respectively., Conclusion: As4S4 induced cellular apoptosis in NB4 cells involves the expression changes of the two genes: the up-regulation of Apaf1 and down-regulation of PNAS-2. The increased expressions of Apaf1 and caspase-9 indicate that As4S4 induced apoptosis in APL cells is through mitochondrial pathway, but not the death-receptor pathway. The role played by PNAS-2 in cellular apoptosis needs to be clarified.

The aim of this study was to establish a simple, convenient and efficient method of producing placental-eluted gamma globulin (PEGG) from human placenta, explore its inhibitory effect on the function of T lymphocyte in vitro and graft-versus-host disease (GVHD) in vivo. PEGG was prepared by elution at acid pH from human placental tissues that were extensively washed. Its effects on T lymphocyte proliferation induced by PHA and mixed lymphocyte reaction (MLR) were analysed by BrdU ELISA, its effect on the CD25 and CD69 expression on T cells was observed by flow cytometry, and the interferon gamma (IFN-gamma) and interleukin-4 (IL-4) quantification in MLR supernatant were assayed by ELISA. A murine GVHD model was established, the effect of PEGG on the manifestation and pathologic change of GVHD and 45-day survival rate were observed. The results showed that considerable level of immunoglobulin could be eluted from placenta at acid PH, of which the main components were IgG checked by SDS-PAGE analysis. In vitro study indicated that PEGG significantly inhibited both the proliferative response of T cells to PHA and the MLR, down-regulated the expression of CD25 and CD69 on T cells stimulated by PHA, and decreased the secretion of IFN-gamma but increased the production of IL-4 in MLR supernatant. In vivo, recipient mice treated with PEGG had a markedly increased survival rate with less histopathological evidence of GVHD. It is concluded that PEGG can inhibit the proliferation and activation of T cells, regulate the direction of T helper cells differentiating towards Th2 type, and effectively prevent GVHD in a murine model. In short, PEGG may be a potent therapeutic agent for GVHD.

The study was aimed to investigate the role and significance of CD123 with other immunological markers in detecting minimal residual disease (MRD) of APL patients. The immunophenotypes of 186 newly diagnosed APL patients and the percentages of cells identical with APL cell immunophenotypes in 20 normal bone marrow samples were analyzed using four-color flow cytometry. MRD in 172 specimens were monitored by mainly using CD34/CD117/CD123/HLA-DR four-color antibody panels, meanwhile 18 specimens were analyzed with the second antibody combination: CD9/CD117/CD34/CD33, simultaneously and the results were compared with real-time PCR. One hundred and sixteen of 172 bone marrow (BM) or peripheral blood (PB) specimens were from follow-up 19 newly diagnosed APL patients and the rest 56 samples were from 47 patients treated 3 to 24 months later. Among them, 117 samples and 55 samples were collected after achieving morphologic complete remission (mCR) and before achieving mCR respectively. The results of immunophenotyping demonstrated that except CD9, CD33 and CD117 were high-expressed and CD34 and HLA-DR were rarely expressed, the CD123 was expressed in 30/30 (100%) APL patients. The percentages of CD117(+)CD34(-)CD123(+)HLA-DR(-) and CD117(+)CD34(-)CD9(+)CD33(+) cells in nucleated cells were 0.066% +/- 0.012% and 0.089% +/- 0.066% in 20 normal bone marrow samples. The median time of achieving morphology complete remission in 19 APL patients was 4 weeks (3 - 6 weeks). The median time of FCM and PCR results turned to be negative in 13 APL patients was 7.5 weeks (5 - 11) and the median time of PCR results turned to be negative in 11 APL patients was 8 weeks (5 - 12). 41/117 (35.04%) samples were MRD positive by FCM after achieving mCR. The ratio of CD117(+)CD34(-)CD123(+)HLA-DR(-) cells was < 5% in 33 specimens, but > 5% in another 8 specimens, their median percentages of CD117(+)CD34(-)CD123(+)HLA-DR(-) cells were 0.48% (range 0.02% - 4.70%) and 9.02% (range 5.26% - 18.14%) respectively. The median relative percentages of CD123(+)HLA-DR(-) cells in CD117(+)CD34(-) population were 63.59% (range 15.11% - 98.36%) and 86.77% (range 63.29% - 92.62%) respectively. In FCM MRD positive samples, 95.9% (93/97) were PCR positive, the false positive rate of FCM and the false negative rate of PCR were 4.1% (4/97) and 8.75% (7/93) respectively. In FCM negative samples, 92% (69/75) were PCR negative and 8% (6/75) were PCR positive. The percentages of CD117(+)CD34(-)CD123(+)HLA-DR(-) cells in 116 consecutive specimens and 117 specimens of mCR were related to PML/RARalpha quantified by real-time PCR (r = 0.824, P < 0.001 and r = 0.754, P < 0.001 respectively). It is concluded that the detection of APL patients by means of two sets of antibody panels is simple and suitable, which is complementary to PCR in monitoring MRD of APL patients.

Objective: To study the cytokeratin 18 and 13 and their gene (CK) expression in post-operation maxillary cyst linings with metaplastic epithelium., Methods: CK expressions were examined with immunohistochemistry in 46 post-operative maxillary cyst (POMC) which were lined with pseudostriated columnar cells only (13 cases), both kinds of columnar and squamous cells (30 cases) and squamous cells only (3 cases)., Results: The expressions of CK8, CK13 and CK18 were observed in 39, 9 and all of the 43 columnar epithelial linings, respectively. Metaplastic squamous epithelia expressed more CK13 and less CK18 and CK8. Of the 33 metaplastic linings, 24 expressed CK8, 23 CK13 and 26 linings expressed CK18. The expression of CK13- and CK18-mRNA was generally correlated with the protein expression level. By in situ hybridization, CK18-mRNA expression was observed not only in 26 metaplastic linings which were positive for CK18 protein but also in five of the seven metaplastic linings which did not express CK18 protein. In addition, RT-PCR revealed an expression of CK18-mRNA in all metaplastic squamous linings although the expression level was weaker than that in the columnar epithelial linings. The CK13-mRNA was expressed in a fashion inverse to the CK18-mRNA., Conclusions: These results indicate that CK18-mRNA is preserved through metaplasia although the protein expression decreases and metaplastic squamous cells differentiate with a decrease of CK18 and an increase of CK13 expression.