OBJECTIVETo screen effective sequences of small interfering RNA targeting MDR1 gene in human gastric cancer SGC7901/VCR cells.

METHODSFour siRNAs (MDR1si326, MDR1si1513, MDR1si2631 and MDR1si3071) targeting MDR1 gene were designed and synthesized by in vitro transcription. The siRNA duplexes were used to transfect into the human gastric cancer SGC7901/VCR cells. The expression level of MDR1 mRNA and P-gp were detected by RT-PCR and Western blotting, respectively. The accumulation of intracellular adriamycin (ADR) was examined by flow cytometry and the cell sensitivity to ADR was demonstrated by MTT.

RESULTSThe SGC7901/VCR cells treated with 4 siRNAs led to reversal effect on multidrug resistance to different extents. Among the SGC7901/VCR cells treated by siRNAs for 48 h, the expression level of MDR1 mRNA in cells of MDR1si326 or MDR1si2631 group (0.42 +/- 0.07 or 0.49 +/- 0.02) was more decreased than that in cells of MDR1si1513 or MDR1si3071 group (P < 0.05). The accumulation of ADR in cells of MDR1si326 group was the most; in cells of MDR1si2631 group, more; in cells of MDR1si3071 group, lower and in cells of MDR1si1513 group, the lowest (P < 0.05). The relative reversal efficiency of cells of MDR1si2631 group to ADR was the highest and in cells of MDR1si326 group, higher (P < 0.05). There was no significant difference in the relative reversal efficiency between the cells of MDR1si1513 and MDR1si3071 groups (P > 0.05). The expression level of P-gp in cells of MDR1si326 group was the lowest among the SGC7901/VCR cells treated by siRNAs for 72 h.

CONCLUSIONThe MDR1si326 with most, MDR1si2631 with more, MDR1si3071 with less and MDR1si1513 with least reversal effects on MDR1 gene mediated multidrug resistance were found in the human gastric cancer SGC7901/VCR cells.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; metabolism ; Antibiotics, Antineoplastic ; metabolism ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Doxorubicin ; metabolism ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Genes, MDR ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; pharmacology ; Stomach Neoplasms ; metabolism ; pathology ; Transfection ; Vincristine ; pharmacology

Animals ; Bone and Bones ; cytology ; metabolism ; pathology ; Collagen ; metabolism ; Durapatite ; metabolism ; Female ; Osteoporosis ; metabolism ; pathology ; Ovariectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the clinical efficacy on primary dysmenorrhea treated with isolated herbal moxibustion on Shenque (CV 8).

METHODSOne hundred and two cases were randomized into isolated herbal moxibustion group and western medication group, 51 cases in each one. In isolated herbal moxibustion group, the cases were treated with isolated herbal moxibustion on Shenque (CV 8), and in western medication group, analgesic was administered.

RESULTSIn isolated herbal moxibustion group, 17 cases were cured, 21 cases effective markedly, 9 cases effective and 4 cases failed. The total effective rate was 92.2%. In western medication group, 7 cases were cured, 11 cases effective markedly, 21 cases effective and 12 cases failed. The total effective rate was 76.5%. The comparison of the total effective rates between two groups indicated significant difference in statistics (P < 0.01), in which, the result in isolated herbal moxibustion group was superior to that in western medication group. In comparison of dysmenorrhea score before and after treatment and side effects of two groups, the significant differences presented in statistics, in which, the results in isolated herbal moxibustion group were superior to those in western medication group (all P < 0.01).

CONCLUSIONThe isolated herbal moxibustion on Shenque (CV 8) is superior to oral medication of analgesic for primary dysmenorrhea and it deserves to be promoted due to its advantages on safety, less side effect and good compliance of patients.

Acupuncture Points ; Adolescent ; Adult ; Dysmenorrhea ; therapy ; Female ; Humans ; Moxibustion ; Young Adult

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.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; analysis ; B-Lymphocytes ; immunology ; Burkitt Lymphoma ; classification ; immunology ; pathology ; Cell Lineage ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; Neprilysin ; analysis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; classification ; immunology ; pathology

OBJECTIVETo compare the leukemia-associated immunophenotypes (LAIP) in patients with B lineage acute lymphoblastic leukemia (B-ALL) at diagnosis and relapse, and investigate its implications for minimal residual disease (MRD) detection.

METHODSThe immunophenotype of leukemia cells from 410 newly diagnosed and 6 relapsed patients with B-ALL were detected by four to six antibody combination, mainly CD34/CD10/CD45/CD19 of 4-color CD45/SSC gating flow cytometry (FCM).

RESULTSThe proportion of CD45 under-expressed or negative in relapsed patients was much higher than that in newly diagnosed patients, being 69.2% and 37.8% respectively. Immunophenotypic changes occurred in 9 relapsed patients (including 8 hematological relapse and 1 central nerves system relapse) when analyzed by paired samples analysis at diagnosis vs at relapse: 4 cases showed CD45 down-modulation and 2 up-modulation; 4 CD34 down-modulation and 2 CD10 up-modulation, while the expression of CD19 remained no change. MRD was observed in all 7 cases of hematological relapse 2 - 4 months before relapse, and the immunophenotype of MRD cells was the same as that in relapse.

CONCLUSIONA high frequency of immunophenotypic changes occurred at relapse and even in MRD before relapse, however the accuracy of MRD monitoring seemed not affected by the FCM strategy used in this investigation.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD19 ; immunology ; Antigens, CD34 ; immunology ; Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; methods ; Leukocyte Common Antigens ; immunology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; immunology ; pathology ; Neprilysin ; immunology ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology ; pathology ; Recurrence ; Young Adult

OBJECTIVETo evaluate the clinical significance for minimal residual disease (MRD) detection by 4 color flow cytometry in B lineage acute lymphoblastic leukemia (B-ALL).

METHODSMRD was analyzed and followed up by using two panels of 4 color antibodies, mainly CD34/CD10/CD45/CD19, in 671 consecutive bone marrow specimens and 1 cerebrospinal fluid from 98 B-ALL patients. In 26 cases of them the immunophenotyping informations at diagnosis were not available.

RESULTSOf 671 bone marrow samples, 579 were MRD negative with leukemic cells below 0.0001 and 93 were MRD positive with leukemic cells over 0.0001. Of 93 MRD positive samples, leukemic cells below 0.05 were found in 64 bone marrow samples, meanwhile in the other 29 samples leukemic cells were over 0.05. Twenty patients relapsed, 19 were bone marrow relapse and one center nerves system. Fifteen of them were found MRD positive 7 - 17 weeks before relapse including 6 patients having no immunophenotyping data at diagnosis. The percentages of leukemia cells in these 15 patients were all over 0.0001. Two relapsed patients were MRD negative in 3 and 9 months before relapse, respectively. Two relapsed after MRD monitoring stopped. If MRD level was > 0.0001 at the end of induction chemotherapy and 12 weeks of treatment, the rate of relapse was 50% (6/12), while, it was 7.5% (3/40) in MRD negative patients (P = 0.000).

CONCLUSIONRelapses can be predicted by MRD monitoring, if MRD was positive in the early phase of treatment, the risk of relapse was higher. Based on the characteristics of B cells ontogeny, MRD detection can be done independently of immunophenotypic information at diagnosis.

Acute Disease ; Adolescent ; Adult ; Aged ; Antigens, CD19 ; immunology ; Antigens, CD34 ; immunology ; Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Follow-Up Studies ; Humans ; Immunophenotyping ; Leukocyte Common Antigens ; immunology ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; immunology ; Neprilysin ; immunology ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology ; Young Adult

This study was purposed to investigate the effect of adenovirus-mediated transfer of PDCD5 gene on apoptosis of K562 cells induced by etoposide. Recombinant adenovirus PDCD5 (Ad-PDCD5), control vectors Ad-null and Ad-eGFP were constructed by AdMax vector system respectively. After K562 cells were transfected by Ad-PDCD5, Ad-null or Ad-eGFP with different multiplicity of infection (MOI), the expression level of the PDCD5 gene was examined by RQ-RT-PCR assay. The effects of etoposide in combination with Ad-PDCD5 on the proliferation and apoptosis of K562 cells were measured by using MTT assay and flow cytometry with Annexin-V-FITC/PI dual labeling technique, respectively. The results showed that the transfection efficiencies of Ad-eGFP in K562, Jurkat and CEM cells ranged from 60% to 86%. Expression level of PDCD5 gene in K562 cells was evidently increased following transfection with Ad-PDCD5. The Ad-PDCD5 synergistically enhanced the apoptotic percentage of K562 cells induced by VP-16, as compared with that of Ad-null + VP16 and VP-16 alone respectively. It is concluded that Ad-PDCD5 may be a potential agent for enhancing the chemotherapy effect.
Adenoviridae ; genetics ; metabolism ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; Etoposide ; pharmacology ; Humans ; K562 Cells ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Transfection

OBJECTIVETo evaluate the method and value of detecting bone marrow minimal residual disease (MRD) in B lineage acute lymphoblastic leukemia (B-ALL) by multiparameter flow cytometry (FCM).

METHODSThe FCM immunophenotyping of B-lymphocyte precursors in regenerating stage and MRD was analyzed by using two sets of 4-color antibody panels, including mainly CD34, CD10, CD45, CD19, in 26 regenerating bone marrow samples after chemotherapy and 297 consecutive bone marrow specimens from 50 patients with B-ALL, respectively. The immunophenotype of leukemia cells of B-ALL was also detected by four to six antibodies combination of 4-color CD45/SSC gating FCM. The CD19, CD10, CD34 fluorescence intensity of B-cell precursors in normal and leukemic bone marrow was compared by relatively quantitative method.

RESULTSTwelve patients were MRD positive (MRD(+)) among 50 patients during MRD monitoring, the percentages of residual leukemia cells were 0.06% to 7.73%. Eleven cases displayed CD45, CD34, CD19, CD10 antigens aberrant expression. Five patients were bone marrow relapsed and 4 of them were MRD (+) 7-17 weeks before relapse. The percentages of leukemia cells in all the 4 patients were over 0.1%. Only one patient relapsed with MRD negative. The regenerated B precursors could be divided into 3 stages according to the expression of CD45, CD34, CD19, CD10, CD22, and CD20 antigens. Abnormal expression of CD45, CD34, CD19, and CD10 were detected in 71.8% of 213 patients with B-ALL, the percentage of only aberrant expression of CD38 and myeloid antigen was 8.1%.

CONCLUSIONDetection of MRD by multiparameter flow cytometry is a rapid, quantitative and sensitive approach, and has a higher predictability for relapse.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; methods ; Leukemia, B-Cell ; diagnosis ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis

OBJECTIVETo ascertain the causation of a pregnant woman with undefined pneumonia reported from the People's Hospital of Tongling city in Anhui province on November 2005.

METHODSEpidemiological and clinical information of the case was collected from the keypersons close to the case and referring to the medical record. A medical observation was carried out on the close contacts of the case and sick or dead poultry. Tracheal aspirates being collected were tested by both RT-PCR and real-time PCR to detect viral nucleic acids of A/H5N1, and were inoculated into special pathogen free (SPF) embryonated hens' eggs.

RESULTSThe pregnant woman was found to have been contacted with the sick/dead poultry directly on the 4th day before onset of illness. All the 122 close contacts were healthy after a 10-day medical observation. The major clinical features of the case were viral pneumonia with rapidly developed leukopenia and lymphopenia. The progress to acute respiratory distress syndrome and multiple organ dysfunction syndromes was found at clinical presentation. HA and NA gene of A/H5N1 virus were positive. The 8 gene fragments of A/Anhui/1/2005 (H5N1) isolated from the tracheal aspirates had not carried genes from a human virus through reassortment, and the receptor-binding site of the hemagglutinin was polybasic cleavage site.

CONCLUSIONThis was the first documented case of H5N1 infection in pregnant woman. The immunotolerant state of pregnancy might have predisposed to the fatal outcome of the patient.

Adult ; China ; Fatal Outcome ; Female ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza, Human ; complications ; pathology ; Multiple Organ Failure ; Pneumonia ; virology ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy Complications, Infectious ; virology ; Respiratory Distress Syndrome, Adult ; Trachea ; virology

One new sorbicillin derivative, 2-deoxy-sohirnone C (1), one new diketopiperazine alkaloid, 5S-hydroxynorvaline-S-Ile (2), and two naturally occurring diketopiperazines, 3S-hydroxylcyclo(S-Pro-S-Phe) (3) and cyclo(S-Phe-S-Gln) (4), together with three known compounds were isolated from the Chinese mangrove endophytic fungus Penicillium sp. GD6. Their structures were determined on the basis of extensive spectroscopic analyses and by comparison with literature data. The absolute configuration of 3-hydroxyl moiety in 3 was determined by Mosher's method, while the absolute stereochemistry of 2 and 4 was established by comparison with the CD spectra of natural and synthesized diketopiperazines. Compound 1 showed moderate antibacterial activity against Methicillin-resistant Staphylococcus aureus with a MIC value of 80 μg·mL.
Alkaloids ; chemistry ; isolation & purification ; Anti-Bacterial Agents ; chemistry ; isolation & purification ; pharmacology ; China ; Circular Dichroism ; Diketopiperazines ; chemistry ; isolation & purification ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Microbial Sensitivity Tests ; Molecular Structure ; Nuclear Magnetic Resonance, Biomolecular ; Penicillium ; chemistry ; Resorcinols ; chemistry ; isolation & purification ; pharmacology ; Rhizophoraceae ; microbiology ; Wetlands