Salmonella enterica serovar Cerro 87, which was isolated from a commercial egg-producing farm, has a phosphorothioated DNA backbone resulting DNA degradation(Dnd) during the pulsed field gel electrophoresis (PFGE) process. In this research, a gene deletion mutant XTG103 was engineered with the entire dnd gene cluster knocked out by double crossover using vector pKOV-kan, and lost Dnd phenotype corre- spondingly. We regulated the DNA phosphorothioation by heterogenous expression of dnd gene cluster with an isopropyl ?-D-1-thiogalactopyranoside (IPTG) inducible promoter PlacZ.

OBJECTIVETo investigate the efficacy and adverse reaction of bortezomib plus chemotherapy with or without stem cell transplantation (SCT) for treatment of multiple myeloma (MM).

METHODSThirty-one MM patients were treated with bortezomib plus dexamethasone or thalidomide or DTPACE, followed by SCT. Response to bortezomib was evaluated according to the European Blood and Marrow Transplantation (EBMT) criteria. Adverse events were graded according to the WHO criteria.

RESULTS1) 5 discontinued the bortezomib therapy because of acute renal failure or acute tumor lysis syndrome and 3 died. In 26 evaluable patients received 99 courses of therapy. The overall response rate (ORR) to bortezomib was 80.8%, and was 100.0% in 15 newly diagnosed patients and 54.6% in 11 relapsed/refractory patients. All of the 6 newly diagnosed patients treated with bortezomib plus DTPACE followed by SCT achieved CR. 2) In 7 newly diagnosed patients completed 8 cycles bortezomib treatment, the diseases were improved more and more with the courses of treatment. Chromosome 13 deletion did not exert a negative impact on response. 3) 6 of 7 patients completed 8 cycles treatment without SCT relapsed in 1-3 month after discontinued therapy; only 1 of 6 such patients received SCT relapsed with the rest keeping on CR at 6-11 month follow-up. 4) The most common adverse events were 1-2 grade and tolerable 3 patients had to reduce the bortezomib dosage because of peripheral neuropathy or sinus bradycardia.

CONCLUSIONBortezomib in combination with chemotherapy with or without SCT is an effective therapy with manageable toxicities for MM patients.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Boronic Acids ; administration & dosage ; Bortezomib ; Dexamethasone ; administration & dosage ; Female ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; surgery ; therapy ; Pyrazines ; administration & dosage ; Thalidomide ; administration & dosage ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo determine the incidence and clinical significance of chromosome 13q14 deletion in multiple myeloma (MM).

METHODSBone marrow samples were collected from 132 newly diagnosed MM patients referred to our hospital. Interphase fluorescence in situ hybridization (i-FISH) combined with magnetic activated cell sorting (MACS) were performed on chromosome 13q14 (RB-1).

RESULTS(1) i-FISH was used to investigate CD138-enriched bone marrow MM cells and revealed a 13q14 deletion rate of 51.5% (68/132), while conventional cytogenetic (CC) analysis revealed 13q deletions/monosomy 13 (Δ13) only of 5.0%(6/120). (2) Univariate analysis showed that 13q14 deletion rate by i-FISH > 25%, bone marrow plasma cells > 50%, ISS stage and β(2)-MG ≥ 5.5 mg/L were associated with shorter overall survival (OS). Multivariate analysis revealed that 13q14 deletion rate by i-FISH > 25% was an independent unfavorable factor (P = 0.042). (3) Patients treated with bortezomib had a much better response than those treated with traditional chemotherapy (P = 0.001). There was no significant difference in OS between patients received bortezomib with and without 13q14 deletion (P > 0.05), indicating that bortezomib could reverse the poor prognosis of 13q14 deletion.

CONCLUSION(1) i-FISH followed CD138 cell sorting appears to be a highly sensitive method for detecting 13q14 deletion. (2) 13q14 deletion rate by i-FISH > 25% is an independent unfavorable factor. (3) Bortezomib could reverse the poor prognosis of 13q14 deletion.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Boronic Acids ; therapeutic use ; Bortezomib ; Chromosome Deletion ; Chromosome Disorders ; Chromosomes, Human, Pair 13 ; Female ; Flow Cytometry ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; drug therapy ; genetics ; pathology ; Prognosis ; Pyrazines ; therapeutic use

OBJECTIVETo study the characteristics of amplitude integrated electroencephalography (aEEG) in preterm infants and changes of maturation with gestational age.

METHODSaEEG monitoring was done within 3 days of age with domestically produced digital aEEG set (CFM3000). Duration of each recording was at least 4 hours. The continuity, sleep-wake cycle, voltage and bandwidth of all aEEG tracing were analyzed.

RESULTSThe percent of continuity background increased from 30% of 28 weeks to 85.7% of 36 weeks (χ(2) = 28.2, P = 0.026); the percent of mature sleep-wake cycle increased from 10% of 28 weeks to 100% of 36 weeks (χ(2) = 192.4, P < 0.01). Low bound voltage increased with gestational age, from (6.8 ± 1.7) µV (28 w) to 9.7 - 10.1 µV (35 - 36 w) (F = 11.4, P < 0.01). Bandwidth of the narrow band decreases gradually with gestational age, from 1.45 cm (28 w) to (0.86 ± 0.24) cm (36 w) (F = 8.731, P < 0.01). The correlation coefficient for continuity, sleep-wake cycle, low bound voltage and bandwidth of narrow band, and total scores were 0.32, 0.81, 0.38, 0.55 and 0.78 respectively (P < 0.05).

CONCLUSIONThe older the gestational age of infants at birth, the more mature the aEEG pattern, manifested as increased continuity and sleep-wake cycle, the higher low bound voltage and more narrowed bandwidth with increased gestational age.

Age Factors ; Electroencephalography ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; physiology ; Male

OBJECTIVETo verify applicability of the International Staging System (ISS) for multiple myeloma (MM) to 112 Chinese MM patients and compare ISS with Durie-Salmon (DS) and Intergroup Francophone du Myeloma (IFM) staging system in predicting prognosis.

METHODS112 previously untreated MM patients in Blood Diseases Hospital of CAMS were analyzed according to ISS retrospectively.

RESULTS1) Serum beta2-microglobulin (beta2-MG) > or = 3.5 mg/L was an independent adverse prognostic factor for overall survival (OS), and serum albumin <35 g/L predicted for time to progression (TTP), 2) In the 58 cases having cytogenetic data, chromosome 13 aberration (Delta 13) was the only independent adverse prognostic factor for OS; 3) Factors significantly related to serum beta2-MG were serum creatinine, 24h urinary protein,body mass index (BMI) and performance status (PS); and those related to serum albumin were hemoglobin level, percentage of bone marrow plasma cells, lactate dehydrogenase(LDN), fever, PS, class of M-protein, serum phosphorus and BMI; 4) All traditional prognostic factors had no statistical difference between ISS stage II and III excepting for serum beta2-MG and creatinine, and 5/6 Delta 13 patients were classified to ISS stage II; 5) The median OS of ISS stage I, II, III were 69, 23 and 26 months (m) respectively, being no statistical difference between stage II and III; for DS system, 89.5% of patients were classified in stage III, being no statistical difference for OS between the stage I/II and III; while for IFM system, the median OS of low-, intermediate- and high-risk group were 69, 40 and 8 months respectively, being statistically different between high-risk and intermediate/ low-risk groups.

CONCLUSIONSFrom the result of our limited analysis, the staging of ISS II and III seems unsuitable for Chinese MM patients. The IFM staging system ,which incorporates delta 13, is more effective than ISS, and DS staging system in predicting prognosis.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; pathology ; Neoplasm Staging ; Prognosis ; Retrospective Studies

OBJECTIVETo establish a new function method for the analysis of a-fetoprotein (AFP) and beta-hCG in testicular tumors.

METHODSWe reexamined the serum levels of AFP and beta-hCG after radical orchiectomy, and calculated the measured coordinate, with the abscissa representing the number of the half-lives of tumor markers, and the ordinate representing the measured value of tumor markers. Referring to the measured value of tumor markers before surgery as a, the number of half-lives as x, and their theoretical value over a period of x elimination half-lives as y (logarithm to the base 2 of y), we calculated the predicted coordinate according to the formula y = log2(a/2x) ==> x + y = log2a (function 1). Then we assessed tumor residue and metastasis by analyzing the relationship between the measured and predicted coordinates.

RESULTSThe pathological examination of case 1 revealed a germ cell tumor of a mixed histological pattern of syncytiotrophoblast and yolk sac tumor. The measured coordinates of AFP and beta-hCG were (2.22, 6.21) and (10, 8.38), and the predicted coordinates (2.22, 6.34) and (10, 4.41) , indicating the elimination of the yolk sac tumor and metastasis of the syncytiotrophoblast tumor. Case 2 demonstrated the mixed pathological nature of teratocarcinoma and yolk sac tumor. The measured coordinates of AFP and beta-hCG were (2.67, -1.03) and (12, -3.32), and the predicted coordinates (2.67, 1.41) and (12, -5.80). But the review times of AFP and beta-hCG were out of the effective range of half-lives, with the measured values below the normal, which suggested no tumor residue or metastasis. Case 3 was found to be embryonal carcinoma. The measured coordinate of AFP was (0.22, 9.25) , and the predicted coordinate (0.22, 9.55) , indicating the elimination of tumor.

CONCLUSIONThe change of the tumor markers predicted by the function method coincided with the natural course of disease in the three cases. The coincidence of the measured with the predicted coordinate after radical orchiectomy indicates no metastasis, while their disagreement suggests possible residue and metastasis of the tumor.

Adult ; Biomarkers, Tumor ; blood ; Chorionic Gonadotropin, beta Subunit, Human ; blood ; Humans ; Male ; Models, Statistical ; Orchiectomy ; Testicular Neoplasms ; metabolism ; pathology ; alpha-Fetoproteins ; analysis

OBJECTIVETo investigate serum adiponectin level in patients with Alzheimer's disease (AD) and its correlation with the patients' cognitive function.

METHODSThis case-control study was conducted in 90 patients with a highly probable diagnosis ofAD, who were divided into mild, moderate and severe group saccording to the MMSE score. Ninety healthy subjects matched for age and gender with the AD patients were selected as the control group. The serum levels ofadiponectin in the participants were detected using enzyme-linked immunosorbent assay.

RESULTSSerum adiponectin level was significantly lower in the AD group than in the control group (P<0.05). Of the 3 subgroups of the AD patients, the moderate and severe AD groups showed significantly lower serum adiponectin level sthan the control group (P<0.05), but the difference in adiponectin levels was not significant between the mild AD group and the control group (P>0.05); serum adiponectin levels also differed significantly among the 3 subgroups of AD patients (P<0.05). Serum adiponectin level was positively correlated with the MMSE score in the AD patients (r=0.683, P<0.001).

CONCLUSIONSerum adiponectin levels are reduced in AD patients and associated with the degree of cognitive impairment.

Adiponectin ; blood ; Alzheimer Disease ; blood ; Case-Control Studies ; Cognition ; Cognitive Dysfunction ; blood ; Enzyme-Linked Immunosorbent Assay ; Humans

OBJECTIVETo discuss the enzyme linked immune spot test (ELISPOT) detected the cellular immune response induced by human Bocavirus (HBoV) VP2 virus-like particles (VLPs).

METHODSAfter immunized by HBoV VP2 VLPs, the specific cellular immune response in mice were detected by ELISPOT assay, observe the ELISPOT results at the conditions of different polypeptide stimulate, different cell culture time, different cell concentration and different specific stimulus peptide concentration, then screening the right ELISPOT experimental conditions and establish the ELISPOT method.

RESULTSThe spots induced by HBoV1 VLPs immunized mice spleen lymphocytes stimulate with polypeptide P3 (GYIPIENEL) and P5 (LYQMPFFLL)were 233 spots/10(6) cells and 157 spots/10(6) cells,spots induced by HBoV2 VLPs immunized mice spleen lymphocytes stimulate with polypeptide P8 (GYIPVIHEL) were 113 spots/10(6) cells; 24 hours is the best time for culture, at this time HBoV1 and HBoV2 groups specificity secretion IFN-gamma ratio were 232 spots/10(6) cells and 119/10(6) cells; Best concentration of mice spleen lymphocyte is 5 x 10(5), right now HBoV1 and HBoV2 group specificity secretion IFN-gamma ratio were 232 spots/10(6) cells and 108/10(6) cells; Best concentration of polypeptides is 10 microg/ml, HBoV1 and HBoV2 group specificity secretion IFN -gamma ratio were 233 spots/10(6) cells and 96/10(6) cells.

CONCLUSIONSHBoV1 and HBoV2 specificT-cell epitope in BABL/c mice were P3, P5 (HBoV1) and P8 (HBoV2). The best experiment condition were: cell cultivated for 24 h, cells concentration for 5 x 10(5) cells/well, stimulating polyperides concentration for 10 microg/ml, it can use to study the cellular immune induced by HBoV in mice.

Amino Acid Sequence ; Animals ; Enzyme-Linked Immunospot Assay ; methods ; Epitopes, T-Lymphocyte ; Human bocavirus ; immunology ; Immunity, Cellular ; Interferon-gamma ; biosynthesis ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Virion ; immunology

OBJECTIVESTo isolate cells of interest from heterogeneous tissue blocks to obtain accurate representations of molecular alterations acquired by neoplastic cells so as to meet the demands of further study on gene expression patterns of the esophageal carcinoma (EC) evolution.

METHODSBlocks of EC were stored at -70 degrees C as close as possible to the time of surgical resection. The tissue block was embedded in OCT and frozen sections of 35 microns in thickness were cut in a cryostat under strict RNAse-free conditions. Individual frozen sections were mounted on plain glass slides and 30-gauge needle attached to a 1 ml syringe was used to microdissect defined cells in the sections. The procured cells were used for total RNA extraction.

RESULTSAn optimized protocol of manual microdissection was developed successfully whereby regions with an area as small as 1/25 mm2 could be accurately dissected. The RNA recovered from procured cells was of high quality suitable for subsequent applications of molecular analysis as assessed of 18S and 28S rRNAs by electrophoresis on agarose gel.

CONCLUSIONSIt is believed that manual microdissection is capable to procure defined cell populations from complex primary tissues, thus allowing investigation of tissue-, cell-, and function-specific gene expression patterns. The technique is simple, easy to perform, versatile, and of particular usefulness when laser capture microdissection (LCM) is practically unavailable.

Cell Separation ; Electrophoresis, Agar Gel ; Esophageal Neoplasms ; genetics ; pathology ; Gene Expression Regulation, Neoplastic ; Genetic Techniques ; Microdissection ; methods ; Neoplasm Staging ; RNA, Neoplasm ; analysis ; isolation & purification

OBJECTIVETo assess the prognostic value of biological features and therapy-related factors in multiple myeloma (MM).

METHODS123 patients with newly diagnosed MM between January 1998 and May 2005 were enrolled in this retrospective study. Biological features at presentation and therapy-related factors were analysed. The overall survival (OS) and time to progression (TTP) were estimated by Kaplan-Meier analysis and the distribution of OS and TTP were compared using log-rank test. Cox regression was used to identify the independent prognostic factors.

RESULTS(1) The univariate analysis indicated that more immature plasma cells in bone marrow biopsy, C-reactive protein >8. Omg/L, CD117 expression, serum beta2-microglobulin (beta2-MG) (3.5 approximately 5.5 mg/L), abnormal cytogenetics aberration of chromosome 13 (Delta13), hypodiploid, poor response to chemotherapy, interferon(IFN) therapy less than 6 months were associated with shorter OS(P <0.05). Lytic bone lesions at presentation, more immature plasma cells in bone marrow biopsy, serum beta2-MG (3.5 approximately 5.5 mg/L), poor response to chemotherapy, and IFN therapy less than 6 months as well as abnormal cytogenetics, hypodiploid and Delta13 were associated with shorter TTP (P <0.05). (2) Multivariable COX analysis indicated IFN therapy more than 6 months was a protective factor for OS and TTP, and more immature plasma cells in bone marrow biopsy was an independent poor prognostic factor for TTP.

CONCLUSIONThe morphology of myeloma cells is useful for assessing the prognosis. And IFN therapy more than 6 months could lengthen OS and TTP.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; pathology ; therapy ; Prognosis ; Retrospective Studies ; Risk Factors ; Survival Analysis