OBJECTIVETo investigate whether the presence of structured CagA proteins in Western- and Eastern-type Helicobacter pylori (H. pylori) induces different incidences of gastric diseases.

METHODSCagA and phosphorylated CagA were expressed in AGS gastric epithelial cells infected with wild type and mutant strains. The ability of individual CagA was determined by immunoprecipitation and Western blot assay. Morphological changes of these cells were observed under microscope to evaluate the appearance of elongation hummingbird phenotype.

RESULTSThe sizes of CagA proteins in different strains were different, and no phosphorylated CagA proteins were detected in wild-type strains. Meanwhile, the kinetics of CagA status in AGS infected with H. pylori was detected. The molecular weight of phosphorylated CagA with the same size of CagA proteins in H. pylori was different in infections with different wild-type strains. CagA and phosphorylated CagA increased in a time-dependent manner after the infection. The hummingbird phenotype with H. pylori for time-course was observed under microscope. Instead of HPK5 strain, the wild-type 26695 strain induced hummingbird phenotype in a time-dependent manner.

CONCLUSIONTranslocation and phosphorylation of CagA are necessary, but not sufficient, for the induction of hummingbird phenotype in AGS cells.

Amino Acid Sequence ; Antigens, Bacterial ; metabolism ; Bacterial Proteins ; metabolism ; Cell Line ; Helicobacter Infections ; metabolism ; microbiology ; Helicobacter pylori ; Humans ; Interleukin-1 ; genetics ; metabolism ; Protein Transport

Alanine Transaminase ; blood ; DNA, Viral ; blood ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B, Chronic ; virology ; Humans ; Male

Hainantoxin-IV (HNTX-IV) purified from the venom of the spider Selenocosmia hainana is a potent antagonist that acts on tetrodotoxin-sensitive (TrX-S) sodium channels. It is a 35-residue polypeptide and includes three disulfide bridges. In order to investigate the structure-function relationship of HNTX-IV, two mutants (S12A-HNTX-IV and R29A-HNTX-IV) of HNTX-TV in which Ser12 and Arg29 were replaced by Ala respectively, were synthesized by solid-phase Fmoc chemistry, followed by oxidative refolding of purified peptides under the optimal conditions. The synthetic mutants were analyzed by MALDI-TOF mass spectrometry, nuclear magnetic resonance spectroscopy (NMR) and electrophysiological experiments for molecular weight, conformation and physiological activity, respectively. The results show that the mutants and native HNTX-IV (nHNTX-IV) have almost identical three-dimensional structures. The bioactivity level of S12A-HNTX-IV is also about the same as that of nHNTX-IV, suggesting that Ser12 does not play any important role for the bioactivity of this toxin. The bioactivity of R29A-HNTX-IV is reduced by at last 155 times, indicating that Arg29 is a key residue relative to the bioactivity of HNTX-IV. It is presumed that the decrease in activity of R29A-HNTX-IV is due to the changes of the property in the binding site rather than the change in the basic conformation of the molecule.
Amino Acid Substitution ; Animals ; Mutation ; Sodium Channel Blockers ; Sodium Channels ; drug effects ; physiology ; Spider Venoms ; chemical synthesis ; genetics ; Structure-Activity Relationship ; Tetrodotoxin ; pharmacology

OBJECTIVETo compare the difference in clinical efficacy on lumber disc herniation (LDH) treated with Yaoyangguan (GV 3) between mild moxibustion under thermosensitive condition and that under non-thermo-sensitive condition.

METHODSFifty-seven LDH patients were selected as the study objects. Mild moxibustion at Yaoyangguan (GV 3) was applied for 45 min each time. Additionally, the conventional acupuncture was given, once a day, for 20 days. At the end of treatment, two groups were classified. A thermosensitive acupoint group (thermosensitive group) was composed of the cases with thermo-sensitization at Yaoyangguan (GV 3) and presenting for > or =4 times in the entire treatment. A non-thermosensitive acupoint group (tranquilization group) was composed of the cases without thermo-sensitization or the frequency of thermo-sensitization <4 times in the entire treatment. The modified Japanese orthopedics association scoring system (M-JOA) was adopted to observe the cases before and after treatment and 6 months after treatment in the two groups. The efficacy was compared between the two groups.

RESULTSAfter treatment and in 6 months after treatment, the score of M-JOA was apparently reduced as compared with that before treatment in the two groups (all P<0.05). The score reducing in the thermosensitization group was more obvious than that in the tranquilization group (both P<0.01). After treatment and in 6 months after treatment, the curative rate and remarkably effective rate were 89.7% (26/29) and 79.3% (23/29), which was better than 71.4% (20/28) and 60.7% (17/28, both P<0.05) in the tranquilization group separately.

CONCLUSIONMild moxibustion at acupoint under thermosensitive condition achieves the better short-term and long-term effects as compared with that under non-thermosensitive condition.

Acupuncture Points ; Adult ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome

Kv4.3 channel is present in many mammalian tissues, predominantly in the heart and central nervous system. Its currents are transient, characterized by rapid activation and inactivation. In the hearts of most mammals, it is responsible for repolarization of the action potential of ventricular myocytes and is important in the regulation of the heart rate. Because of its central role in this important physiological process, Kv4.3 channel is a promising target for anti-arrhythmic drug development. Jingzhaotoxin-V (JZTX-V) is a novel peptide neurotoxin isolated from the venom of the spider Chilobrachys jingzhao. Whole-cell patch clamp recording showed that it partly blocked the transient outward potassium channels in dorsal root ganglion neurons of adult rats with an IC(50) value of 52.3 nmol/L. To investigate the effect of JZTX-V on Kv4.3 channel, JZTX-V was synthesized using the solid-phase chemical synthesis and separated by reverse phase high performance liquid chromatography (HPLC). The purity was tested by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MOLDI-TOF mass spectrometry). Two-electrode voltage-clamp technique was used to characterize the action of JZTX-V on Kv4.3 channels expressed in Xenopus laevis oocytes. As a result, JZTX-V displayed fast kinetics of inhibition and recovery from inactivation. Furthermore, it could inhibit Kv4.3 channel current in a time- and concentration-dependent manner with an IC(50) value of 425.1 nmol/L. The application of JZTX-V affected the activation and inactivation characteristics of Kv4.3 channel and caused a shift of the current-voltage relationship curve and the steady-state inactivation curve to depolarizing direction by approximately 29 mV and 10 mV, respectively. So we deduced that JZTX-V is a gating modifier toxin of Kv4.3 channel. Present findings should be helpful to develop JZTX-V into a molecular probe and drug candidate targeting to Kv4.3 channel in the myocardium.
Animals ; Ganglia, Spinal ; cytology ; Neurons ; drug effects ; Neurotoxins ; pharmacology ; Oocytes ; Patch-Clamp Techniques ; Peptides ; pharmacology ; Potassium Channel Blockers ; pharmacology ; Rats ; Shal Potassium Channels ; metabolism ; Spider Venoms ; pharmacology ; Xenopus laevis

OBJECTIVETo investigate the mechanism and the suppression effect of human cytomegalovirus (HCMV) on hematopoietic system.

METHODSSemi-solid culture system was used to observe the effect of HCMV AD169 strain on colony forming unit granulocyte/macrophage (CFU-GM), CFU-erythroid (CFU-E), CFU-multipotent (CFU-Mix) and CFU-megakaryocyte (CFU-MK) growth. The techniques of in situ polymerase chain reaction (IS-PCR) and polymerase chain reaction (PCR) were used to demonstrate the existence of HCMV DNA in the colony cells of cultured CFU-GM, CFU-Mix, CFU-MK and CFU-E, respectively. The immediate early antigen (IEA) mRNA in CFU-MK and late antigen (LA) mRNA in CFU-E were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). HCMV early protein P52 was detected with immunohistochemical technique.

RESULTSHCMV AD169 suppressed the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK in vitro significantly (P < 0.05). The suppression was dose-dependent. HCMV DNA was successfully detected in CFU-GM, CFU-Mix, CFU-MK colony cells from viral infection groups by IS-PCR, and was detected in CFU-E by PCR, while it was negative in blank control or mock control groups. CFU-MK colony cells expressed HCMV IEA mRNA with the size of 340 bp in virus infection groups of 10(3) plague forming unit (PFU), 10(4) PFU and 10(5) PFU, respectively. The HCMV LA mRNA was detected by RT-PCR and was 263 bp long in positive control group of HCMV-infected human embryonic fibroblasts. The expression of HCMV LA mRNA in CFU-E was negative. The early protein P52 of HCMV in 10(4) PFU group was also identified by immunohistochemical staining.

CONCLUSIONHCMV AD169 strains inhibited the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK by the infection of the hematopoietic progenitors. HCMV might cause the suppression of hematopoiesis by direct infection, which is thought to be one of the reasons of HCMV infection associated with thrombocytopenia, neutropenia and anemia.

Colony-Forming Units Assay ; Cytomegalovirus ; genetics ; DNA, Viral ; genetics ; Erythrocytes ; virology ; Hematopoietic System ; cytology ; virology ; Humans ; Megakaryocytes ; virology ; Multipotent Stem Cells ; virology ; Polymerase Chain Reaction

OBJECTIVETo evaluate the clinical therapeutic value of (188)Re-HEDP combined with pamidronate in breast cancer with bone metastasis.

METHODSForty-eight patients with breast cancer with multi-bone metastases were randomly divided into three groups:15 patients received (188)Re-HEDP (group A), 15 patients received pamidronate (group B) and 18 patients were treated by (188)Re-HEDP plus pamidronate (group C).

RESULTSThe overall pain relief rate was 73.3%, 80.0%, 100.0% in groups A, B and C. The response rate of bone metastasis was 40.0%, 33.3%, 66.7% in groups A, B and C respectively. The therapeutic effect of group C was better than those of groups A and B (P < 0.05), without any significance in the difference (P > 0.05).

CONCLUSIONThe therapeutic effect of (188)Re-HEDP combined with pamidronate for breast cancer with bone metastasis is remarkable in bone pain relief and bone metastasis control, which is better than either (188)Re-HEDP or pamidronate alone.

Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Bone Neoplasms ; complications ; secondary ; therapy ; Breast Neoplasms ; drug therapy ; pathology ; radiotherapy ; Carcinoembryonic Antigen ; blood ; Combined Modality Therapy ; Diphosphonates ; therapeutic use ; Etidronic Acid ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Organometallic Compounds ; therapeutic use ; Pain ; etiology ; Pain Management ; Radioisotopes ; therapeutic use ; Rhenium ; therapeutic use

OBJECTIVETo investigate the feasibility of 9Tc(m)-HL91 imaging in prediction of 34 radiotherapy sensitivity of naqsopharyngeal cancer( NPC) and its relationship with prognosis.

METHODSpatients with NPC confirmed by pathology, staging from II-IVa, underwent 99Tc(m)-HL91 SPECT imaging one week before radiotherapy. 18 of them received adjuvant chemotherapy. The hypoxia in primary nasopharyngeal lesions and cervical lymph node metastases were calculated semi-quantitatively, and compared with clinical findings in medium-term therapy at 4 months and 1 year post therapy.

RESULTS(1) There was no significant relationship between the total preliminary curative effect of adjuvant chemotherapy and the degree of nasopharyngeal lesion hypoxia (T/Mu, gamma = -0.394, P = 0.145; T/ Ce gamma = -0.510, P = 0.052). But there was a significant difference between the partial curative effect group and significant curative effect group. (2) The degree of NPC regression in the medium-term radiotherapy group was negatively correlated with the degree of hypoxia (T/Mu, gamma = -0.602; T/Ce, gamma = -0.643, P < 0.01). (3) 23 patients had good local control except one case with lung and bone metastasis 4 months post-therapy. The lesions disappeared or not developed in 6 patients (T/Mu 1.30 +/- 0.23, T/Ce 3.61 +/- 0.84). Two patients at stage III and IVa relapsed (T/Mu were 1.40 and 1.27, respectively; T/Ce were 4.10 and 3.85, respectively), there was no significant difference. (4) The degree of lymph node hypoxia had no correlation with the curative effect on medium-term radiotherapy.

CONCLUSION99 Tc(m)-HL91 hypoxia imaging may predict sensitivity to radiotherapy in patients with NPC, with a potential help to carry out individual therapy. However, further investigation is needed to ascertain whether it could predict the long-time curative effect on NPC radiotherapy.

Adult ; Aged ; Chemotherapy, Adjuvant ; Female ; Follow-Up Studies ; Humans ; Hypoxia ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnostic imaging ; drug therapy ; radiotherapy ; Neoplasm Staging ; Organotechnetium Compounds ; Oximes ; Particle Accelerators ; Preoperative Care ; methods ; Prognosis ; Radiotherapy, High-Energy ; Remission Induction ; Reproducibility of Results ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo explore the protective effect of tanshinone II A on lipopolysaccharide (LPS)-induced lung injury in rats, and possible mechanism.

METHODSLPS (O(111): B4) was used to produce a rat model of acute lung injury. Sprague-Dawley rats were randomly divided into 3 groups (8 in each group): the control group, the model group (ALI group), and the tanshinone II A treatment group. Expression of adhesion molecule CD18 on the surface of polymorphonuclear neutrophil (PMNCD18) in venous white blood cells (WBC), and changes in coagulation-anticoagulant indexes were measured 6 h after injection of LPS or normal saline. Changes in malondialdehyde (MDA) content, wet and dry weight (W/D) ratio and morphometry of pulmonary tissue as well as PMN sequestration in the lung were also measured.

RESULTS(1) When compared with the control group, expression of PMNCD18 and MDA content were enhanced in the ALI group with a hypercoagulable state (all P<0.01) and an increased W/D ratio (P<0.05). Histopathological morphometry in the lung tissue showed higher PMN sequestration, wider alveolar septa; and lower alveolar volume density (V(V)) and alveolar surface density (S(V)), showing significant difference (P<0.01). (2) When compared with the ALI group, the expression of PMN-CD18, MDA content, and W/D ratio were all lower in Tanshinone II A treatment group (P<0.05) with ameliorated coagulation abnormality (P<0.01). Histopathological morphometry in the lung tissue showed a decrease in the PMN sequestration and the width of alveolar septa (both P<0.01), and an increase in the V(V) and S(V) (P<0.05, P<0.01).

CONCLUSIONTan II A plays a protective role in LPS-induced lung injury in rats through improving hypercoagulating state, decreasing PMN-CD18 expression and alleviating migration, reducing lipid peroxidation and alleviating pathological changes.

Animals ; Blood Coagulation ; drug effects ; CD18 Antigens ; analysis ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; pharmacology ; Female ; Lipopolysaccharides ; toxicity ; Lung ; drug effects ; pathology ; Male ; Malondialdehyde ; analysis ; Phenanthrenes ; pharmacology ; Rats ; Rats, Sprague-Dawley

This study was aimed to investigate the expression and role of 14-3-3ζ in the AML cell lines: sensitive HL-60 and drug-resistant HL-60/VCR cells. Semi-quantitative RT-PCR and Western blot were respectively used to examine the expression of mdr1 mRNA and Pgp in AML cell lines to validate the results of microarray. Western blot was performed to investigate the expression of Pgp, 14-3-3ζ, and anti-apoptosis protein BCL-2, MCL-1 proteins. Immunofluorescence assay was used to detect the subcellular location of 14-3-3ζ protein in HL-60 and HL-60/VCR cells by laser scanning confocal microscopy. Transduction with siRNA was used to silence 14-3-3ζ in AML cell lines. Cell count method and flow cytometry of cell cycle were used to analyze the changes of growth of AML cells. The results found that mdr1 mRNA and Pgp did not expressed in HL-60 cells, but significantly overexpressed in HL-60/VCR cells. Except 14-3-3σ, the expression of other subtypes of 14-3-3 was higher in HL-60/VCR cells than that in HL-60 cells, especially 14-3-3ζ. The higher expression of 14-3-3ζ, BCL-2, MCL-1 protein was observed in HL-60/VCR cells than that in HL-60 cells. These results were same results from gene chip. It was also noticed that 14-3-3ζ was located in the cytoplasma and nuclei of AML cell lines, especially over-expressed in HL-60/VCR cells. Furthermore, suppression of 14-3-3ζ by RNA interference resulted in inhibition of the proliferation of AML cells with decreased protein expression of BCL-2 and MCL-1, especially in HL-60/VCR cells. It is concluded that 14-3-3ζ plays an important role in proliferation of AML cells and associates with BCL-2 and MCL-1 expression. These results suggested that development of therapy targeting 14-3-3ζ may provide novel, effective strategies for refractory and relapsed AML.
14-3-3 Proteins ; metabolism ; ATP Binding Cassette Transporter, Sub-Family B ; metabolism ; Apoptosis ; Cell Proliferation ; HL-60 Cells ; metabolism ; Humans ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism