Objective To investigate risk factors for Pseudomonas aeruginosa infected AECOPD patients and the signifi-cance of immune regulation .Methods The clinical pharmacist is involved in the administration and guardianship of immuno-compromised patients with recurrent AECOPD .The clinical pharmacist provides recommendations and theoretical support in antimicrobial drug selection and regulating immunity .Results Clinical pharmacist gave rationalized medication recommenda-tions .It made anti-infective programs more reasonable and effective .Conclusion By assisting physicians in formulating regi-mens ,clinical pharmacists can provide more optimal individualized treatment for patients .

OBJECTIVETo investigate the quantities of monocyte-derived dendritic cell precursors (pDC1) and plasmacytoid dendritic cell precursors (pDC2) in peripheral blood mononuclear cells (PBMC) of severe aplastic anemia (SAA) patients before and after immune suppressive therapy (IST), the ratio of the pDC1 to pDC2, and the expression of co-stimulating molecules (CD80, CD86, CD40) on dendritic cells (DC) and B cells in SAA patients.

METHODSBy means of three color monoclonal antibody labeling technology, the quantities and ratio of pDC1 and pDC2 in PBMC were detected in 26 SAA patients at active phase, 13 at recovery phase and 15 normal controls respectively. The aforementioned parameters of 10 SAA patients were tested before and 2 months after IST. The expression of CD80, CD86 and CD40 on DC and B lymphocytes were detected in 16 SAA patients and 15 normal controls.

RESULTSThe percentages of pDC1 and the ratio of pDC1/pDC2 of controls were (0.41 +/- 0.05)% and 1.58 +/- 0.18 respectively, and those of SAA patients at active phase were (0.67 +/- 0.13)% and 2.70 +/- 0.32 respectively, [pDC1 (P < 0.05); pDC1/ pDC2 ratio (P < 0.01)]. The aforementioned parameters in convalescent SAA patients decreased to (0.43 +/- 0.10)%, and 1.78 +/- 0.36 respectively, being no difference from those of normal controls. The percentages of pDC1 and pDC2 in 10 SAA patients were (0.87 +/- 0.31)%, and (0.35 +/- 0.09)%, before IST, and (0.24 +/- 0.09)%, (0.14 +/- 0.04)%, after IST, being significantly decreased (P < 0.05). The percentages of CD86 expression on DC of controls was (11.97 +/- 4.31)%, and that of SAA patients was (29.84 +/- 3.02) % (P < 0.05). The percentages of CD80, CD40 and CD86 expression on lymphocytes of controls were (2.57 +/- 0.44)%, (7.34 +/- 1.22)% and (1.86 +/- 1.11)%, respectively, and those of SAA patients were (5.17 +/- 0.68)%, (8.85 +/- 2.94)% and (5.98 +/- 0.96)% respectively (P < 0.05, P < 0.01). The percentage of CD86 expression on B lymphocytes in controls was 8.04 +/- 0.66%, and in SAA patients was (20.46 +/- 2.78)%, (P < 0.05).

CONCLUSIONThe pDC subtypes were abnormal and the percentage of pDC1 is increased in SAA patients, which are associated with stage of this disease. DC and B Lymphocytes in SAA patients upregulated expression of costimulatory molecules (CD86) which cause the T lymphocyte abnormally activated.

Adolescent ; Adult ; Anemia, Aplastic ; immunology ; B-Lymphocytes ; immunology ; metabolism ; B7-1 Antigen ; blood ; B7-2 Antigen ; blood ; CD40 Antigens ; blood ; Case-Control Studies ; Child ; Convalescence ; Dendritic Cells ; immunology ; metabolism ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged

OBJECTIVETo clone human canstatin gene and express its recombinant protein.

METHODSThe total RNA was extracted from human placenta. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and transformed into E.coli DH5alpha through electroporation. The gene was sequenced by the Sanger Dideoxy-mediated chain-termination method, and then the canstatin cDNA was cloned into the BamHI and HindIII sites of plasmid pET-22b (+) and transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b-Dgalactopyranoside (IPTG).

RESULTSThe extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting products were cloned into pUCm-T vectors, and then were transformed into E.coli DHSa. After an over night culture, both blue and white colonies were found on the agar plate. Six white colonies were selected and cut by BamHI and HindIII. The plasmids DNA in one white colony showed one band near the location of primary plasmid after digested by BamHI and two bands near the locations of primary plasmid and objective gene fragment after digested by HindIII. The cloned gene in this white colony was sequenced and demonstrated to have the same sequence as that of canstatin gene in GenBank. Then canstatin cDNA was cut down from pUCm-T with BamHI and HindIII and ligated into the vector pET-22b (+). The resultant plasmid pET-22b (+)/canstatin was then transformed into E.coli BL21. White colonies were found on LB agar plate. Seven of them were selected and their plasmids were digested with both BamHI and HindIII. After electrophoresis, all selected colonies showed two specific bands, one was found near the location of primary plasmids, and the other near that of objective gene fragment. After IPTG induction, there was a new protein band about Mr 24 000 on SDS-PAGE. As estimated by densitometry, the percentage of the expressed product over total bacterial proteins was 18.2%, 18.8%, 23.0% and 23.4%, respectively, 1, 2, 3, and 4 hours after induction.

CONCLUSIONHuman canstatin gene was successfully cloned and its recombinant proteins were expressed in this study.

Base Sequence ; Cloning, Molecular ; methods ; Collagen Type IV ; biosynthesis ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; In Vitro Techniques ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Aim To assess the value of the administration of adenosine-5'-triphosphate (ATP) during sinus rhythm for noninvasive diagnosis of AV node dual pathways(AVNDP) and abolition or modification of the slow pathway (SP) after radiofrequency(RFCA) in patients with inducible sustained AVNRT. Methods Incremental doses of ATP were intravenously administrated during sinus rhythm to patients with spontaneous or inducible sustained AVNRT(study group, n=45)and to patients with no evidence of AVNDP or inducible AVNRT (control group, n=37) until ECG signs of AVNDP( 50 ms increase or decrease in P-R interval in two consecutive beats, or occurrence of AVNRT) or second-degree AV block were observed. Results Four patients (two in study patients and two in control patients) could not complete the trial and were excluded from analysis. AVNDP was observed by ATP in 36(84%) study patients, whereas it was diagnosed by electrophysiology criteria in 38(88%) patients. AVNDP was observed only in 1(3%) control patient. AVNDP by ATP test was disappeared in 18(90%) of 20 patients who underwent SP abolition and in 3(38%) of 8 patients who underwent SP modification. Conclusion ATP test during sinus rhythm enables noninvasive diagnosis of AVNDP in a high percentage of patients with inducible AVNRT and reliably confirms the results of RFCA of the SP.

OBJECTIVE: To determine the damage of different types of floods on the residents health. METHODS: The methods of standard mortality rate (SMR) and years of potential life lost (YPLL) were used to analyze the death of sample residents from flood areas in Dongting Lake in Hunan province. RESULTS: The order of death causes in the soaked area, the collapsed area and the non-flood area was the same. But the mortality rates of residents injury, poisoning and malignant neoplasm diseases in the soaked area and the collapsed area were higher than those of non-flood area. The resident standard rates of years of potential life lost (SYPLL) in the soaked area and the collapsed area were higher than that of the controls, especially in the age group of 30 to 45. The flood-attributed SYPLL in the male was higher than that of the female. CONCLUSION Flood actually affected the health of residents. The more serious the flood is, the worse the effect is. It is very important to decrease the resident mortality rate of the injury, poisoning and malignant neoplasm, and to pay attention to protect people of 30 to 45 years old in flood areas.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Disasters ; Female ; Humans ; Infant ; Life Tables ; Male ; Middle Aged ; Mortality

OBJECTIVETo investigate the abnormal hematopoietic clone burden of the patients with myelodysplastic syndromes (MDS) and its clinical implication.

METHODSThe ratio of the metaphase with abnormal karyotypes to the total was regarded as the index of MDS clonal burden. Thirteen parameters were assayed and the correlations between these parameters and MDS clone burden were analysed.

RESULTSThe clonal burden of MDS patients was (67.4 +/- 36.2)%. It correlated positively with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = -0.445, P < 0.05). The number of blasts, hemoglobin and erythrocytes in high clonal burden (>50%) and low clonal burden (< or = 50%) groups were significantly different (P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (274.18 +/-71.85) x 10(6)/L and (454.82 +/- 205.88) x 10(6)/L (P < 0.05) respectively. CD8+ T lymphocytes between MDS patients and normal controls had no difference. The serum level of IL-2 of MDS patients and normal control groups were (6.29 +/- 3.58) g/L and (3.11 +/- 1.40) microg/L (P < 0.05) respectively; but no difference in the serum level of TNF between MDS and control groups. The ratio of CD4+ to CD8+ in high clonal burden patients was 1.90 + 0.52, and in low clonal burden patients was 0.97 +/- 0.44 (P < 0.05).

CONCLUSIONThe clonal burden and deficient T cell immunity are the indicators for predicting MDS patients clinical progression.

Adolescent ; Adult ; Aged ; Bone Marrow Cells ; pathology ; Chromosome Aberrations ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; immunology ; pathology ; T-Lymphocytes ; immunology

OBJECTIVETo analyse the proportion of hepatitis associated aplastic anemia (HAAA) in severe aplastic anemia (SAA) and its clinical features of HAAA.

METHODSAll newly diagnosed SAA cases in our department in the recent 5 years were analyzed. A case-control study was undertaken to investigate the differences of clinical and laboratory features between HAAA and non-hepatitis associated SAA (non-HASAA) patients.

RESULTSThe proportion of HAAA in SAA was 3.3%. There was no significant difference in PB cell counts, bone marrow hematopoiesis status and the amount of blood transfusion between HAAA and non-HASAA patients. Sera from 13 patients with HAAA were tested for antibodies to hepatitis viruses A, B, and C and hepatitis B surface antigen. Twelve (92.3%) of them had negative serologic results for the tests and only one (7.7%) had a positive result for HBsAg and HBeAg. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were decreased prior to the diagnosis in twelve (92.3%) of the 13 HAAA patients. The percentage of CD4(+) cells in HAAA patients was significantly lower than that in non-HASAA patients (P < 0.05). HAAA patients had higher percentages of CD8(+) cells (P < 0.05) and lower ratios of CD4(+)/CD8(+) (P < 0.05). The early infection rate of the HAAA patients was significantly higher than that of non-HASAA patients (84.6% vs 42.3%, P < 0.05), with different mortalities (61.5% vs 15.4%, P < 0.05). The 2-year survival rate of HAAA patients was significantly lower than that of non-HASAA patients (16.6% vs 83.2%, P < 0.01).

CONCLUSIONThe proportion of HAAA in SAA was 3.3%. Most of HAAA were associated with non-A, non-B and non-C hepatitis virus. Compared with that of non-HASAA, the abnormality of T cell immunity of HAAA was more severe, with a higher frequency of early infection and a higher mortality rate.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; complications ; pathology ; Case-Control Studies ; Female ; Follow-Up Studies ; Hepacivirus ; immunology ; Hepatitis A Antibodies ; blood ; Hepatitis A virus ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B virus ; immunology ; Hepatitis C Antibodies ; blood ; Hepatitis, Viral, Human ; blood ; complications ; virology ; Humans ; Male

OBJECTIVETo study the response of hematopoietic cells (HSC) to granulocyte colony stimulating factor (G-CSF) in paroxysmal nocturnal hemoglobinuria (PNH) patients.

METHODS(1) Bone marrow mononuclear cells (BMMNC) from 17 PNH patients and 12 normal subjects were inoculated into semisolid culture media containing or not G-CSF (50 ng/ml). The cluster/colony forming unit-granulocyte/monocyte (CFU/cFU-GM) were counted and compared. (2) BMMNC of 20 PNH patients and 12 normal controls were triply stained for CD34, CD59 and G-CSF receptor CD114/stem cell factor receptor (C-KIT) CD117 and assessed by FCM. The CD34(+) cells were identified as CD34(+)/CD59(+) and CD34(+)/CD59(-). Percentage of CD114 and CD117 expression in each cell population was calculated.

RESULTS(1) PNH cFU-GM without G-CSF were (112.41 +/- 22.74)/10(5) BMMNC, while with G-CSF: (133.82 +/- 25.85)/10(5) BMMNC and normal cFU-GM were (190.33 +/- 36.05)/10(5) BMMNC, (309.42 +/- 92.94)/10(5) BMMNC, respectively. Whether with or without G-CSF, PNH BMMNC formed less cFU-GM than control did, both of the two kinds of BMMNC responded to G-CSF well (P < 0.05), but the increment of PNH cFU-GM yields was less than that of the normal control (P < 0.05). CFU-GM yields of PNH BMMNC without G-CSF were (24.29 +/- 9.05)/10(5) BMMNC, with G-CSF were (27.53 +/- 10.65)/10(5) BMMNC, while normal control were (77.42 +/- 36.01)/10(5) BMMNC and (98.00 +/- 43.14)/10(5) BMMNC, respectively. Whether with or without G-CSF, PNH BMMNC showed less CFU-GM yields than that of control (P < 0.05). (2) The percentage of CD114 positive cells in PNH CD34(+)CD59(+) BMMNC was (73.34 +/- 29.40)% and that in PNH CD34(+)CD59(-) BMMNC and in control CD34(+)CD59(+) BMMNC were (32.70 +/- 6.89)% and (58.52 +/- 29.99)%, respectively. The percentage of CD114 expression in PNH CD34(+) CD59(-) BMMNC was less than that in the other two groups (P < 0.05). The percentages of CD117 positivities on the PNH CD34(+)CD59(+) BMMNC were (76.90 +/- 22.08)%, PNH CD34(+) CD59(-) (36.03 +/- 7.69)% and control CD34(+) CD59(+) (80.28 +/- 13.36)%, respectively (P < 0.01).

CONCLUSIONIn vitro, BMMNC of normal control grow better, and respond better to G-CSF than PNH BMMNC do. PNH CD34(+)CD59(-) BMMNC express less G-CSF receptor and C-KIT than PNH CD34(+)CD59(+) and normal CD34(+)CD59(+) BMMNC do, which may be the reason that abnormal PNH clone grow worse than the normal clones do.

Adolescent ; Adult ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; drug effects ; metabolism ; CD59 Antigens ; metabolism ; Cells, Cultured ; Colony-Forming Units Assay ; Female ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Cell Growth Factors ; metabolism ; Hemoglobinuria, Paroxysmal ; blood ; pathology ; Humans ; Male ; Middle Aged ; Proto-Oncogene Proteins c-kit ; metabolism ; Receptors, Granulocyte Colony-Stimulating Factor ; metabolism ; Young Adult

OBJECTIVETo investigate the quantities of bone marrow CD5+ B lymphocytes in the patients with autoimmune hemocytopenia and the relationship between quantities of CD5+ B lymphocytes and clinical or laboratorial parameters.

METHODSQuantities of CD5+ B lymphocytes in the bone marrow of 14 patients with autoimmune hemolytic anemia (AIHA) or Evans syndrome, 22 immunorelated pancytopenia (IRP) patients, and 10 normal controls were assayed by flow cytometry. The correlation between their clinical or laboratorial parameters and CD5+ B lymphocytes was analyzed.

RESULTSThe quantity of CD5+ B lymphocytes of AIHA/Evans syndrome (34.64% +/- 19.81%) or IRP patients (35.81% +/- 16.83%) was significantly higher than that of normal controls (12.00% +/- 1.97%, P < 0.05). However, there was no significant difference between AIHA/Evans syndrome and IRP patients (P > 0.05). In all hemocytopenic patients, the quantity of bone marrow CD5+ B lymphocytes showed significantly negative correlation with serum complement C3 level (r = -0.416, P < 0.05). In the patients with AIHA/Evans syndrome, the quantity of bone marrow CD5+ B lymphocytes showed significantly positive correlation with serum indirect bilirubin level (r = 1.00, P < 0.05). In Evans syndrome patients, the quantity of CD5+ B lymphocytes in bone marrow showed significantly positive correlation with platelet-associated immunoglobulin G (r = 0.761, P < 0.05) and platelet-associated immunoglobulin M ( r = 0.925, P < 0.05). The quantity of CD5+ B lymphocytes in bone marrow of all hemocytopenic patients showed significantly negative correlation with treatment response (tau-b = -0.289, P < 0.05) , but had no correlation with colony forming unit-erythroid (r = -0.205, P > 0.05) or colony forming unit-granulocyte-macrophage colonies (r = -0.214, P > 0.05).

CONCLUSIONSThe quantity of bone marrow CD5+ B lymphocytes in the patients with autoimmune hemocytopenia significantly increases and is correlated with disease severity and clinical response, which suggest that CD5+ B lymphocytes might play an important role in the pathogenesis of autoimmune hemocytopenia.

Anemia, Hemolytic, Autoimmune ; drug therapy ; immunology ; Autoimmune Diseases ; drug therapy ; immunology ; B-Lymphocytes ; classification ; immunology ; Cyclosporine ; therapeutic use ; Drug Therapy, Combination ; Flow Cytometry ; Glucocorticoids ; therapeutic use ; Humans

OBJECTIVETo investigate the role of the burden of abnormal hematopoietic clone in the development of myelodysplastic syndromes (MDS).

METHODSThe ratio of the bone marrow cells with abnormal chromosomes to the total counted bone marrow cells was regarded as the index of MDS clone burden. The disease severity related parameters including white blood cell count, hemoglobin, platelet count, lactate dehydrogenase level, bone marrow blast, myeloid differentiation index, micromegakaryocyte, transfusion, interleukin-2, tumor necrosis factor (TNF), CD4+ and CD8+ T cells of MDS patients were assayed, and the correlations between those parameters and MDS clone burden were also analyzed.

RESULTSThe clone burden of MDS patients was 67.4% +/- 36.2%. MDS clone burden positively correlated with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = -0.445, P < 0.05). The number of blasts, hemoglobin, and erythrocytes in high clone burden (> 50%) and low clone burden ( < or = 50%) groups were 7.78% +/- 5.51% and 3.45% +/- 3.34%, 56.06 +/- 14.28 g/L and 76.40 +/- 24.44 g/L, (1.82 +/- 0.48) x 10(12)/L and (2.32 +/- 0.66) x 10(12)/L, respectively (all P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (0.274 +/- 0.719) x 10(9)/L and (0.455 +/- 0.206) x 10(9)/L, respectively (P < 0.05). CD8+ T lymphocytes of MDS patients and normal controls were (0.240 +/- 0.150) x 10(9)/L and (0.305 +/- 0.145) x 10(9)/L, respectively. The serum level of interleukin-2 of MDS patients (6.29 +/- 3.58 ng/mL) was significantly higher than normal control (3.11 +/- 1.40 ng/mL, P < 0.05). The serum level of TNF of MDS patients and normal control group were 2.42 +/- 1.79 ng/mL and 1.68 +/- 0.69 ng/mL, respectively. The ratio of CD4 to CD8 was higher in high clone burden MDS patients (1.90 +/- 0.52) than that in low clone burden patients (0.97 +/- 0.44, P < 0.05).

CONCLUSIONThe quantitive clonal karyotype abnormalities and deficient T cell immunity are important parameters for evaluating MDS severity and predicting its progression.

Adolescent ; Adult ; Aged ; Bone Marrow Cells ; pathology ; Case-Control Studies ; Chromosome Aberrations ; Female ; Hematopoiesis ; genetics ; Hematopoietic Stem Cells ; pathology ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; genetics ; pathology ; Neoplastic Stem Cells ; pathology ; Polycythemia ; genetics ; pathology ; T-Lymphocyte Subsets ; pathology ; Young Adult