Objective To observe the effects of chloroquine phosphate on apoptosis of leukemic cell line U937, and investigate whether chloroquine phosphate induces leukemic cell apoptosis by normalizing protein PNAS-2's abnormal subcellular location. Methods Chloroquine phosphate of different concentrations were added into culture fluid of leukemic cell line U937 at logarithmic phase. MTr was used to measure cell proliferation, flow cytometry and laser confocal microscopy were applied to detect cell apoptosis, and immunofluorescence technology was employed to observe the effects of chloroquine phosphate on the changes of subcellular location of protein PNAS-2. Results Apoptosis of leukemic cell line U937 was significantly induced by 50 μg/mL chloroquine phosphate, and subcellular location of protein PNAS-2 was changed. Conclusion Chlorequine phosphate can induce apoptosis of leukemic cell line U937, and the mechanism may be related to the normalization of PNAS-2's abnormal subcellular location in U937 cell line. Chloroquine phosphate has the potential to be used in leukemic therapy.

The experiment's aim was to optimize the processing technology of Xanthii Fructus which through comparing the difference of UPLC fingerprint and contents of toxicity ingredient in water extract of 16 batches of processed sample. The determination condition of UPLC chromatographic and contents of toxicity ingredient were as follows. UPLC chromatographic: ACQUITY BEH C18 column (2.1 mm x 100 mm, 1.7 microm) eluted with the mobile phases of acetonitrile and 0.1% phosphoric acidwater in gradient mode, the flow rate was 0.25 mL x min(-1) and the detection wavelength was set at 327 nm. Contents of toxicity ingredient: Agilent TC-C18 column (4.6 mm x 250 mm, 5 microm), mobile phase was methanol-0.01 mol x L(-1) sodium dihydrogen phosphate (35: 65), flow rate was 1.0 mL x min(-1), and detection wavelength was 203 nm. The chromatographic fingerprints 16 batches of samples were analyzed in using the similarity evaluation system of chromatographic, fingerprint of traditional Chinese medicine, SPSS16.0 and SIMCA13.0 software, respectively. The similarity degrees of the 16 batches samples were more than 0.97, all the samples were classified into four categories, and the PCA showed that the peak area of chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeic acid were significantly effect index in fingerprint of processed Xanthii Fructus sample. The outcome of determination showed that the toxicity ingredient contents of all samples reduced significantly after processing. This method can be used in optimizing the processing technology of Xanthii Fructus.
Caffeic Acids ; analysis ; toxicity ; Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; toxicity ; Quinic Acid ; analogs & derivatives ; analysis ; toxicity ; Xanthium ; chemistry ; classification

Objective To establish a method of labeling human mesenchymal stem cells (MSCs) with PKH26 in vitro.Methods MSCs were cultured and labeled with PKH26 according to the manufacturer's instruction.The growth,fluorescence intensity and serial subcuhivation of labeled MSCs were analyzed with the confocal laser microscope and the flow cytometry.The biological characteristics of labeled MSCs were investigated by RT-PCR.Results The labeled MSCs appeared red fluorescence and the labeling rate was 100 percent.During serial subcuhivation of labeled MSC from passage 1 to passage 7,the fluorescence intensity and the labeling rate of MSCs were gradually decreased.The biological features such as morphology,growth,expression level of nucleostemin and GAPDH gene and capability of differentiation into osteoblast in vitro were not affected by labeling.Conclusion Labeling the human MSCs with PKH26 is an effective and practical method,which can be used as an important tool in the study on the homing, plasticity and transplantation of MSCs.

Objective To ascertain the characteristics of the parturient women in the district, the requirement and satisfaction for postpartum visits, and to explore the present situation of postpartum visits in the district, providing theoretical basis for the formulation and improvement of targeted work mechanism for postpartum visits. Methods A total of 540 parturients were selected to complete the questionnaire survey according to the proportion of parturients in each community, then statistical analysis of questionnaire information was made. Results Among the 540 parturients, 400 were primiparas and 140 multiparas.More than 94% of the respondents believed that postpartum visits were necessary.The demand for postpartum visits was different between primiparas and multiparas.The primiparas needed more guidance on the nursing of the newborn, and the multiparas wanted to have more concern for postpartum recovery and health care.Respondents had high satisfaction with postpartum visits. Conclusion The form and content of postpartum visits directly affect the satisfaction of puerperants, who need higher quality and personalized services.The professional quality of visitors should be improved by professional training.And the personalized postpartum visiting service should be properly carried out by making full use of modern means of information dissemination.And postpartum psychological health care also needs attention in this regard.

The high expression of tissue factor (TF) is related to the coagulation disorder in acute leukemia. TF in blood circulation is mainly expressed in cells, microparticles (MP) and alternatively spliced human tissue factor (asHTF). To elucidate the role of TF in the coagulation disorder of acute myeloid leukemia (AML), RT-PCR was performed on 6 common AML cell lines NB4, HL-60, Kasumi-1, U937, K562 and THP-1. The results showed that only NB4 and U937 cells expressed baseline full-length TF and asHTF which were proved by sequencing. The flow cytometric detection, TF activity and TF antigen tests in NB4 and U937 cells revealed that the asHTF was expressed in trace amount and almost had no activity, while the TF antigen and activity in microparticles were significantly higher than that in asHTF. It is concluded that asHTF may play an unimportant role in the coagulation disorder of AML. Microparticle associated tissue factor (MP-TF) is the predominant source of TF activity released from AML cells.
Alternative Splicing ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; Thromboplastin ; genetics ; metabolism ; Tumor Cells, Cultured ; U937 Cells

This study was purposed to prepare and primarily identify the specific monoclonal antibodies (McAbs) against the apoptosis related protein PNAS-2 so as to provide the essential tool for study of PNAS-2 function. The McAbs against PNAS-2 were prepared via the immunization of mice, cell fusion and cloning using synthetic peptide of PNAS-2 as immunogen; the specificity, titer and subtype of McAb were detected by Western blot, ELISA and immunofluorescence. The results showed that the stable hybridoma cell line S-31-7 producing McAbs against PNAS-2 protein was successfully obtained. The immunoglobulin of the McAb was identified to be IGg1lambda. The titer of ascetic fluid fled McAb were 1:8,000. A single specific band with 28 kD was shown in Western blot test, and the antigen recognized was present in cell cytoplasm by immunofluorescence. In conclusion, the obtained McAb against PNAS-2 displays strong specificity and high titer, which may be applied to the advanced research on PNAS-2 protein.
Animals ; Antibodies, Monoclonal ; immunology ; isolation & purification ; Antibody Specificity ; immunology ; Apoptosis Regulatory Proteins ; immunology ; Female ; Mice ; Mice, Inbred BALB C

OBJECTIVETo develop a single-tube multiplex polymerase chain reaction (mPCR) technique to detect three common deletional alpha-thalassemias (alpha-Thal) in Chinese, and to perform genetic diagnosis and prenatal diagnosis for an alpha-Thal family from Hebei province, China.

METHODSFourty-two blood samples including samples from one alpha-Thal family from Hebei province were assayed. The mPCR containing 7 primers, gel electrophoresis and DNA sequencing were used for the genetic diagnosis and prenatal diagnosis.

RESULTSThe gene types of the fourty-two DNA samples analyzed by the mPCR-gel electrophoresis technique were in accordance with the results by Southern blot and three separate PCR techniques. A HbH child and a fetus of the alpha-Thal family were diagnosed as--(SEA)/alpha(cs)alpha and alpha alpha/alpha alpha respectively by using the mPCR and DNA sequencing. The result of postnatal analysis of the cord blood was consistent with the prenatal result (alpha alpha/alpha alpha).

CONCLUSIONThe developed mPCR technique can be used for genetic diagnosis and prenatal diagnosis of the 3 deletional alpha-Thal in Chinese.

Adult ; Child, Preschool ; Family Health ; Female ; Fetal Diseases ; diagnosis ; genetics ; Gene Deletion ; Genetic Testing ; Humans ; Male ; Molecular Diagnostic Techniques ; methods ; Point Mutation ; Polymerase Chain Reaction ; methods ; Pregnancy ; Prenatal Diagnosis ; Sequence Analysis, DNA ; alpha-Thalassemia ; diagnosis ; genetics

BACKGROUNDThe degree of pathological microvascular proliferation is an important element in evaluation of the astrocytoma grade. This study was aimed to quantitatively assess the microvascular permeability of brain astrocytoma with the volume transfer constant (K(trans)) and volume of extravascular extracellular space per unit volume of tissue (Ve) from dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) and to evaluate the effectiveness of the K(trans) and Ve in the grading of astrocytoma.

METHODSThe highest values of the K(trans) and Ve of 67 patients with astrocytoma (27 with grade II, 12 with grade III, and 28 with grade IV) were obtained. The comparisons of the differences of the K(trans) and Ve between the different grades were conducted using the Mann-Whitney rank-sum tests. Spearman's rank correlation coefficients were determined between K(trans) values, Ve values and astrocytoma grades. Receiver operating characteristic (ROC) curve analyses were performed to determine the cut-off values for the K(trans) and Ve to distinguish between the different grades of astrocytoma.

RESULTSThere were significant differences (P < 0.001) between the different grades in the K(trans) values and Ve values, except for grades III and IV. The K(trans) values and Ve values were both correlated with astrocytoma grades (both P < 0.001). The ROC curve analyses showed that the cut-off values for the K(trans) and Ve provided the best combination of sensitivity and specificity in distinguishing between grade II and grade III or IV astrocytomas.

CONCLUSIONSDCE-MRI can play an important role in assessing the microvascular permeability and the grading of brain astrocytoma.

Adolescent ; Adult ; Aged ; Astrocytoma ; pathology ; physiopathology ; Brain Neoplasms ; pathology ; physiopathology ; Capillary Permeability ; physiology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the frequency of mitochondrial DNA (mtDNA) D-loop hypervariable region II (HVR II) and hypervariable region III (HVR III) mutations in oral squamous cell carcinoma (OSCC) and their correlation to provide the new targets for the prevention and treatment of OSCC.

METHODSThe D-loop HVR II and HVR III regions of mtDNA in seven cases with OSCC tissues, matched with paracancerous tissues and normal mucosa tissues from the same case, were amplified by polymerase chain raction (PCR), then were detected by direct sequencing to find the mutantsites after the comparison of all sequencing results with the mtDNA Cambridge sequence in the GenBank database.

RESULTS82 (56 species) nucleotide changes, with 51(26 species) nucleotide polymorphism, were found after the comparison of all sequencing results with the mtDNA Cambridge sequence in the GenBank database. 31(30 species) mutations, with 21 located within the HVR II and HVR III regions, were found in 3 tumor tissue samples, their paracancerous and normal mucosa tissue were found more polymorphic changes but no mutation. The mtDNA D-loop HVR II and HVR III regions mutation rate was 42.9% (3/7) in OSCC.

CONCLUSIONThe mtDNA D-loop HVR II and HVR III regions were highly polymorphic and mutable regions in OSCC. It suggested that the D-loop HVR II and HVR III regions of mtDNA might play a significant role in the tumorigenesis of OSCC. It may become new targets for the gene therapy of OSCC by regulating the above indexes.

Carcinoma, Squamous Cell ; DNA, Mitochondrial ; Female ; Humans ; Mouth Neoplasms ; Mutation ; Polymorphism, Genetic

Adult ; Cauda Equina ; Female ; Humans ; Magnetic Resonance Imaging ; Meningeal Neoplasms ; diagnosis ; pathology ; Meningioma ; diagnosis ; pathology ; Spinal Cord Neoplasms ; diagnosis ; pathology