OBJECTIVETo investigate the protective effect of suppressive oligodeoxynucleotides (Sup ODN) on interferon-γ (IFN-γ) and signal transducers and activators of transcription (pSTAT4) expression of Silica-induced pulmonary inflammation in Mice.

METHODSSixty Balb/c mice were randomly divided into 4 groups, normal control group, silicious group, suppressive oligodeoxynucleotides (Sup ODN) group, control oligodeoxynucleotides (Con ODN) group. Except the normal control group injected normal saline, the rest groups were induced by the intratracheal instillation of 0.1 ml (5 g/L) of sterilized silica suspension. Sup ODN group and Con ODN group were treated by i.p. injection of 0.3 ml (1mg/mL) of suppressive or control ODN 3 h before silica administration. After 7 days, the animals were killed and levels of IFN-γ were detected by ELISA. The pathologic changes in lung tissues of mice were observed with HE staining. Expressions of IFN-γ and pSTAT4 in lung tissue were detected with immunohistochemistry and quantified by Image-Pro Plus 7.0.

RESULTSHE staining showed that the lung tissue of silicious group were damaged seriously than Sup ODN group. Compared with the normal control group (serum: (280.1±41.3) pg/ml, lung tissue: (0.249±0.373), IFN-γ increased in silicious group (serum: (886.3±81.7) pg/ml, lung tissue: (0.270±0.300) (P < 0.05). Compared with the normal control group and Con ODN group [(894.5±91.6) pg/ml], IFN-γ in the serum of Sup ODN group decreased significantly (P < 0.01). Compared with the silicious group , IFN-γ in lung tissue decreased in Sup ODN group (0.241±0.250) (P < 0.05). Compared with the normal control group (0.279±0.353), pSTAT4 in lung tissue increased significantly in silicious group (0.313±0.231) (P < 0.01). Compared with the silicious group, pSTAT4 in lung tissue decreased significantly in Sup ODN group (0.269±0.523) (P < 0.01).

CONCLUSIONSup ODN attained protective effect on Silica treated mice by suppressing expression of IFN-γ and pSTAT4.

Animals ; Female ; Inflammation ; metabolism ; Interferon-gamma ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Oligodeoxyribonucleotides ; pharmacology ; Phosphorylation ; STAT4 Transcription Factor ; metabolism ; Silicon Dioxide ; toxicity

OBJECTIVETo observe different effects of moxibustion on extracellular potassium ion in acupoint under physiological and pathological status and provide experimental evidence for exploring action mechanism of moxibustion on acupoint local.

METHODSForty female SD rats were randomly divided into a blank group, a blank-moxibustion group, a model group and a model-moxibustion group, 10 cases in each one. The complete Freund's adjuvant(CFA) was adopted to establish model of adjuvant arthritis (AA) in the model group and model-moxibustion group. No treatment was given in the blank group and model group while moxibustion was applied at "Zusan-li" (ST 36) for 30 min in the blank-moxibustion group and model-moxibustion group. The tissue fluid in "Zusanli" (ST 36) was collected with microdialysis and real-time analyzed by electrolytic analyzer. The change of concentration of potassium ion in "Zusanli" (ST 36) was observed.

RESULTS(1) Under physiological status, the concentration of extracellular potassium ion in the blank group was not changed within 150 min (P > 0.05); before the moxibustion, the concentration of extracellular potassium ion in the blank-moxibustion group was (1.21 +/- 0.31) mmol/L, and after treatment it was gradually increased and reached its peak at (2.38 +/- 0.42) mmol/L after 60 min (P < 0.05), then it was reduced. 150 min after the treatment, concentration of potassium ion was slightly higher than that before moxibustion as well as that in the blank group. The concentration in the blank-moxibustion group at 60 min was statistically significant compared with that in the blank group (P < 0.05). (2) Under pathological status, the concentration of extracellular potassium ion in the model group was not changed within 150 min, differences of which at each time point was not statistically significant (all P > 0.05). Before the moxibustion, the concentration of extracellular potassium ion was (1.09 +/- 0.12) mmol/L in the model-moxibustion group, and it was immediately increased to (1.96 +/- 0.18) mmol/L after moxibustion. 60 min and 90 min after the moxibustion, it still maintained a higher level, which was (1.87 +/- 0.29) mmol/L and (1.59 +/- 0.16) mmol/L respectively (both P < 0.05). The differences of each time point after moxibustion in the model-moxibustion group were statistically significant compared with those in the model group (all P < 0.05).

CONCLUSIONThe moxibustion could increase the concentration of potassium ion in rat's acupoint local under physiological status but time of effect is short; with moxibustion at "Zusanli" (ST 36) under pathological status, the concentration of local potassium ion is obviously increased and maintains for a long time.

Acupuncture Points ; Animals ; Arthritis, Experimental ; metabolism ; therapy ; Disease Models, Animal ; Female ; Humans ; Moxibustion ; Potassium ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore a new access for internal rigid fixation of mandibular mid-and-low condylar fracture.

METHODS16 patients of unilateral mid-and-low condylar fractures were treated with a 2cm mini-retromandibular approach. The subcutaneous tissues superficial to the superficial muscular aponeurotic system (SMAS) were dissected forward that parallel to the masseter muscle fiber bundles, aiming to the fracture. After exposing the fracture, the fracture segments were reduced and fixed under sufficient exposure.

RESULTSCorrect anatomic reduction and occlusion were achieved in all cases. Additionally, all patients showed normal articular function and the surgical scars were barely visible.

CONCLUSIONThe transmasseter approach by retromandibular access is one of the feasible methods for curing mid-and-low condylar fracture, which minimizeing the risk of facial nerve injury and reducing the visible scars.

Adult ; Dental Occlusion ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Mandible ; Mandibular Condyle ; Mandibular Fractures ; Masseter Muscle ; Middle Aged ; Treatment Outcome

OBJECTIVETo determine the relationship between prehospital delay time (PDT) and other associated factors on mortality in patients with acute myocardial infarction.

METHODSWe retrospectively analyzed factors associated with mortality in 580 patients with acute myocardial infarction presented to the Emergency Ward and Emergency Intensive Care Unit (EICU) of Beijing Anzhen Hospital from March 2004 to March 2006 (428 males, average age: 60.7 +/- 12.9 years). The patients were divided to 3 groups according various therapies: thrombolysis, PCI/CABG or symptomatic medication groups.

RESULTSThe median PDT was 130 min. Thrombolysis, PCI/CABG and medical therapy were applied in 122 (21.0%), 266 (45.9%) and 192 (33.1%) patients respectively. PDT was significantly longer in patients receiving medical therapy (290.9 min +/- 3.4 min) compared to patients treated with thrombolysis (104.5 min +/- 2.3 min) and PCI/CABG (119.1 min +/- 2.3 min, all P < 0.05). The overall mortality rate was 5.3% (31/580) and all occurred in patients with medical therapy group mostly due to irreversible ventricular fibrillations. Old age (OR = 1.047, P = 0.004), diabetes mellitus (OR = 2.159, P = 0.02) and PDT (OR = 2.159, P = 0.023) are independent predict factors for mortality.

CONCLUSIONCoronary Revascularisation by thrombolysis, PCI or CABG early post acute myocardial infarction is the key issue for reducing mortality in patients with acute myocardial infarction.

Adult ; Aged ; Aged, 80 and over ; Diagnostic Errors ; Emergency Service, Hospital ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; mortality ; therapy ; Patient Admission ; Prognosis ; Retrospective Studies ; Risk Factors ; Time Factors

OBJECTIVETo investigate the effects of a microfluidic sperm sorter on the routine parameters and DNA integrity of human sperm.

METHODSWe divided 40 semen samples into two aliquots and performed sperm sorting using a self-made polydimethylsiloxane microfluidic sperm sorter and the swim-up method, respectively. Then we evaluated and compared the effects of these two methods on the sperm routine parameters and DNA integrity by computer-assisted sperm analysis and sperm chromatin dispersion test.

RESULTSAfter processing, sperm motility, normal morphology and tail hypoosmotic swelling rate were significantly improved, while sperm DNA damage remarkably decreased (P < 0.01). The microfluidic sperm sorter achieved a significantly lower rate of sperm DNA damage than the swim-up method ([ 8.4 +/- 5.8 ]% vs [16.4 +/- 9.2] %, P < 0.01), but no statistically significant differences were found in all other parameters between the two methods.

CONCLUSIONHigh-quality sperm with less DNA integrity damage could be obtained in sperm sorting with the microfluidic sperm sorter.

Adult ; Cell Separation ; instrumentation ; methods ; DNA ; DNA Damage ; Humans ; Infertility, Male ; genetics ; physiopathology ; Male ; Microfluidics ; Middle Aged ; Protein Array Analysis ; Semen Analysis ; instrumentation ; methods ; Sperm Motility ; Spermatozoa

BACKGROUNDWe determined the diagnosis of hereditary hemorrhagic telangiectasis (HHT) in a suspected HHT family, identified ALK1 gene mutation and established a gene diagnosis method of HHT. The level of related plasma proteins (transforming growth factor beta and thrombomodulin) were also analyzed.

METHODSBleeding history and family history were collected; Dilatant nasal mucosal capillaries in proband were observed under nasal cavity endoscope; exons 3, 7, 8 of ALK1 gene in proband and her family members were amplified with polymerase chain reaction (PCR), and the PCR products were analyzed. Using enzyme-linked immunosorbent assay (ELISA), plasma TGF-beta1 and TGF-beta2 concentrations were measured. Plasma thrombomodulin (TM) level was detected by Western blotting.

RESULTSOf all family members, four had epstaxis, two had evident telangiectases on skin or mucosa. Gene screening results showed that C to T substitution at position 1231 in exon 8 of ALK1 gene (CGG-->TGG) existed in proband, her affected brother and their father. The mutation did not exist in proband's sister-in-law and nephew. Plasma TGF-beta1 concentrations in the affected HHT was 20,538, 17,194, 13,131 pg/ml, while that of normal control and unaffected family members was 15,950, 20,297, 12,836 pg/ml, respectively. Plasma TGF-beta2 in HHT patients was 14,502, 9550, 10,592 and that of normal controls 8579, 20,297, 7680 pg/ml respectively. Level of plasma TM was in HHT subjects significantly lower than in normal subjects.

CONCLUSIONSChinese HHT individuals have mutant ALK1 gene, a C1231T variation on exon 8 of ALK1 is responsible for HHT clinical phenotypes in this family. ALK1 gene analysis, together with special clinical phenotypes and family history, provides a reliable method in diagnosing HHT. In affected HHT subjects, plasma TGFbeta levels were not obviously different from those of normal subject; while plasma TM concentration was significantly lower than that in normal subjects. The significance and mechanism remain to be elucidated.

Activin Receptors, Type I ; genetics ; Activin Receptors, Type II ; Aged ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Telangiectasia, Hereditary Hemorrhagic ; blood ; genetics ; Thrombomodulin ; blood ; Transforming Growth Factor beta ; blood

OBJECTIVETo investigate the functional relations between the putative proteins YpCD1.08, YpCD1.09, YpCD1.16 encoded in pCD1 plasmid of Yersinia pestis and its type III secretion system (T3SS).

METHODSMutants of YpCD1.08, YpCD1.09, YpCD1.16 were constructed using λ-Red recombinant system. The growth curves of the mutant strains cultivated in TMH medium with or without calcium at 26 °C and 37 °C were determined to analyze the low calcium response phenotype. The transcription levels of ΔYpCD1.08, ΔYpCD1.09, ΔYpCD1.16 in Yersinia pestis and the dependence to temperature were determined using real time RT-PCR after cultivation at 26 °C and 37 °C and extraction of RNA. A β-lactamases reporter system was adopted to study the influence of these genes on the translocation of effector YopE of T3SS.

RESULTSWhen grown in TMH medium without calcium at 26 °C and 37 °C, the growth curve of the YpCD1.08, YpCD1.09, YpCD1.16 mutants were similar to that of the wild-type strain, indicating that the low calcium response of all the mutants were normal. The ratios of YpCD1.08, YpCD1.09, YpCD1.16 gene transcriptional level at 37 °C and 26 °C were 2.3 ± 0.3, 2.3 ± 0.5 and 3.2 ± 0.7, respectively, indicating that these genes were transcribed in Yersinia pestis and their transcription regulations showed a temperature-dependence that was consistent with the well established temperature-dependent expression of Yersinia T3SS genes. The β-lactamases reporter assays demonstrated that ΔYpCD1.08 could translocate much higher level of YopE into HeLa cells, since that the light intensity ratio of 477/520 nm at 140 min was 2.5, whereas it was 1.8 for the wild-type strain, and the values in ΔYpCD1.09 and ΔYpCD1.16 were similar to the wild-type strain.

CONCLUSIONYpCD1.08, YpCD1.09, YpCD1.16 gene are likely to be the new members of T3SS, and the putative protein YpCD1.08 could play some roles in YopE secretion and translocation.

Bacterial Outer Membrane Proteins ; secretion ; Bacterial Secretion Systems ; genetics ; Genes, Bacterial ; Plasmids ; Protein Interaction Mapping ; Yersinia pestis ; genetics ; metabolism ; pathogenicity

A little is known about the specific marker on the surface of acute leukemia cells, leading to the lack of the specific diagnosis method for acute leukemia. Therefore, in this study, cell-systematic evolution of ligands by exponential enrichment (cSELEX) was performed to screen the aptamers binding to CD33(+)/CD34(+) cells from the patients with acute myeloblastic leukemia (AML) of M(2) subtype (AML-M₂) so as to provide the basis for finding the specific marker on the surface of AML-M(2) CD33(+)/CD34(+) cells. Firstly, AML-M₂ CD33(+)/CD34(+) cells were sorted and used as targeted cells, and normal CD33(+)/CD34(+)cells were used as counter-targeted cells; the aptamers binding to CD33(+)/CD34(+) cells from patients with AML-M₂ were screened from the single strand deoxyribonucleic acid (ssDNA) library by cSELEX. Subsequently, each aptamer structure was analyzed after cloning and sequencing. The results indicated that after 13 round of screenings, the enrichment of aptamers in the ssDNA library was ranged from 0.7% to 52.9%, and reached steady state at 13th round screening. Sequence analysis for 30 aptamers showed that most of the aptamers born one of the three conserved sequences of CCCCT, CTCTC, and CTCAC. Secondary structure analysis indicated that three different secondary structures existed in these aptamers. It is concluded that the aptamers binding to the AML-M(2) CD33(+)/CD34(+) cells are successfully screened, which lay the basis for further looking for the specific marker on the surface of AML-M₂ CD33(+)/CD34(+) cells, and the molecular diagnosis of the AML-M₂ leukemia.
Antigens, CD ; genetics ; immunology ; Antigens, CD34 ; genetics ; immunology ; Antigens, Differentiation, Myelomonocytic ; genetics ; immunology ; Aptamers, Nucleotide ; metabolism ; Biomarkers ; Flow Cytometry ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; genetics ; immunology ; Nucleic Acid Conformation ; SELEX Aptamer Technique ; Sialic Acid Binding Ig-like Lectin 3

OBJECTIVETo compare the hemodynamic responses to orotracheal intubation using a Shikani Optical Stylet (SOS) laryngoscope or a Macintosh direct laryngoscope (MDLS).

METHODSTotally 41 patients with American Society of Anesthesiologists ASA physical status -aged 20-60 years and scheduled for elective surgery under general anesthesia requiring orotracheal intubation, were randomly allocated to either the SOS group (n=21) or MDLS group (n=20). After an intravenous anesthetic induction the orotracheal intubation was performed using a SOS laryngoscope or a MDLS. Blood pressure and heart rate (HR) were recorded before and after anesthetic induction immediately after intubation, and 5 minutes after intubation. Rate pressure product RPP were calculated.

RESULTSBlood pressures and RPP in both two groups significantly decreased after anesthetic induction (P<0.05) while blood pressures HR, and RPP significantly increased after orotracheal intubation (P<0.05). HR in both groups after intubation were significantly higher than the pre-induction level (P<0.05)and such an increase lasted for 3 min. HR immediately after intubation was also significantly higher in MDLS group than in SOS group (P<0.05); however, such difference was not observed in other time points (P>0.05). In the MDLS group when compared with the occurrence time required for the maximum values of systolic blood pressure (SBP)the occurrence time required for the maximum values of HR after the start of intubation and success of intubation during the observation were significantly delayed (P<0.05). Compared with the MDLS group, the occurrence time required for the maximum values of SBP after the start of intubation and the success of intubation were significantly delayed in the SOS group (P<0.05). The incidences of SBP more than 130% of baseline value and RPP more than 22 000 were not significantly differently(P>0.05). Also, the intubation time was not significantly different (P>0.05).

CONCLUSIONThe hemodynamic responses to orotracheal intubation is milder in SOS laryngoscope than in MDLS.

Adult ; Blood Pressure ; physiology ; Female ; Heart Rate ; physiology ; Hemodynamics ; Humans ; Intubation, Intratracheal ; instrumentation ; methods ; Laryngoscopes ; Male ; Middle Aged ; Young Adult

This study was aimed to investigate the effects of flavonoids of puerarin (PR) on apoptosis of acute promyelocytic leukemia (APL) cell line NB4 cells and its mechanism. The NB4 were treated with PR in vitro, the MTT assay was used to detect the inhibitory effect of PR on cell proliferation. The apoptosis of NB4 cells were detected by flow cytometry labelled with Annexin V/PI. The expressions of pml/rar alpha, bcl-2 and survivin were detected by real time reverse transcription-polymerase chain reaction (real time RT-PCR), the expressions of JNK, p38 MAPK, FasL, caspase 3, caspase 8 were detected by Western blot. The results showed that with the increasing of PR concentrations, the apoptosis rates of NB4 cells were gradually elevated. Simultaneously, the mRNA expression of pml/rar alpha, bcl-2 and survivin decreased, while the protein expression of JNK, FasL, caspase 3 and caspase 8 increased, which presented the positive correlation to PR concentrations. When PR combined with arsenic trioxide (ATO), the expression levels of above mentioned mRNA and protein decreased or increased more significantly. It is concluded that PR can effectively induce the apoptosis of NB4 cells. PR combined with ATO displays synergistic effect. It may be triggered by the activation of JNK signal pathway.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Isoflavones ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Leukemia, Promyelocytic, Acute ; pathology