Objective: To examine the microbial flora within the root canals with failed root canal therapy by PCR test, to identify detection rate of bacteria and the predominant bacteria species, and to determine the association of the various species with clinical features. Methods: Forty roots with failed root canal therapy were selected for this study. According to clinical features the roots were divided into three groups: pain, fistula and symptomless. After removal of the root filling material, the microbial samples were taken from the canal and detected by PCR test. Results: The microbial flora within root canals of teeth with failed root canal therapy was found to be mixed, six kinds of bacteria were detected. Enterococcus faecalis was the most commonly recovered bacteria species. Significant associations were observed between Prevotella nigrescens and pain, while dependablity was observed between Actinomyces israelii and fistula. Conclusion: The main cause of root canal treatment failure is the persistent microorganisms. The composition of bacteria in teeth with failed root canal therapy has its own specificity.

OBJECTIVETo study the protective effects of Ginkgo biloba extract (EGb) on the lung injury of dogs undergoing hypothermic cardiopulmonary bypass (CPB).

METHODSFourteen healthy hybrid dogs were randomly divided into the control group and the EGb group, 7 in each group. EGb (8 mg/kg) was intravenously dripped to dogs in the EGb group before thoracotomy after anesthesia, while equal volume of normal saline was intravenously dripped to those in the control group. The lung tissue was collected after 60-min CPB and 120-min recovery of heart beat. The lung tissues were collected to detect the wet weight-dry weight ratio and the permeability. The contents of malondialdehyde (MDA), the activities of myeloperoxidase (MPO) and total superoxide dismutase (T-SOD) in the lung tissues were detected by biochemical assay. The levels of IL-1beta, IL-8, tumor necrosis factor alpha (TNF-alpha), platelet activating factor (PAF) in the lung tissue were detected by enzyme linked immunosorbent assay (ELISA).

RESULTSCompared with the control group, the wet weight-dry weight ratio was reduced and the permeability of the lung tissue decreased (P < 0.05), the MDA content was reduced, the activity of MPO decreased, and the activity of T-SOD increased (P < 0.05), and the levels of IL-1beta, IL-8, and PAF obviously decreased (P < 0.05).

CONCLUSIONSEGb showed better protective effects on the lung injury of dogs undergoing hypothermic CPB. Its possible mechanisms might be associated with alleviating ischemia-reperfusion injury of in vitro circulation and systemic inflammatory response.

Animals ; Cardiopulmonary Bypass ; adverse effects ; methods ; Dogs ; Ginkgo biloba ; Lung Injury ; etiology ; prevention & control ; Plant Extracts ; pharmacology

OBJECTIVETo evaluate the effects of Ginaton (Ginkgo biloba leaf extract) on the myocardial injury markers (MIMs) during cardiopulmonary bypass (CPB).

METHODSForty patients with congenital heart diseases, scheduled to take atrial septum or ventricular septum repairing operation, were randomly divided into the Ginaton group and the control group, 20 cases in each group. Patients in both groups received St. Thomas' cardioplegic perfusion via radix aortae, while Ginaton (0.5 mg/kg) was added into the perfusion for the Ginton group. Cardiac surgery were started after complete heart arrest. Central venous blood was obtained before and at 0, 6th, 12th, 24th and 48th hour after operation for detection of serum C reaction protein (CRP) by immunoturbidimetry, as well as creation kinase-MB isoenzyme (CK-MB), cardiac troponin T (cTnT) and cardiac troponin I (cTnI) with enzyme-linked immunosorbent assay (ELISA).

RESULTSThere was no difference in serum concentration of CRP, CK-MB, cTnT and cTnI between the two groups before operation (P > 0.05). These indexes increased immediately after operation in both groups ( P < 0.05). They reached the peak value 12 hrs after CPB and reduced to normal level 48 hrs post-operation in the control group, with the value significantly higher than that in the Ginaton group at all the corresponding time points (P < 0.05, or P < 0.01).

CONCLUSIONPerfusion with Ginaton during CPB could significantly decrease the release of MIMs and improve post-CPB cardiac function recovery, exerting favorable myocardium-protective effects.

Biomarkers ; blood ; C-Reactive Protein ; metabolism ; Cardiopulmonary Bypass ; Child ; Child, Preschool ; Creatine Kinase, BB Form ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Female ; Ginkgo biloba ; chemistry ; Heart Defects, Congenital ; surgery ; Humans ; Immunohistochemistry ; Isoenzymes ; blood ; Male ; Myocardial Reperfusion Injury ; blood ; physiopathology ; prevention & control ; Phytotherapy ; Plant Leaves ; chemistry ; Troponin T ; blood

OBJECTIVETo investigate the effects of Ginkgo biloba extract (Ginaton) on bcl-2 and bcl-xL mRNA expression in the myocardium of patients underwent hypothermic cardiopulmonary bypass (CPB).

METHODSThirty congenital heart disease patients were randomly assigned to 2 groups, the control group and the treated group. Patients in both groups received St. Thomas' cardioplegic solution via radix aortae, while Ginaton (0.5 mg/kg) was added in the treated group. Cardiac surgery was started after complete heart arrest. Myocardium was taken before the aorta ascendens was unblocked and mRNA expression of bcl-2 and bcl-xL in the ventricular tissue was detected by RT-PCR.

RESULTSThe gene expressions of bcl-2 and bcl-xL were significantly higher in the treated group than those in the control group (P < 0.05).

CONCLUSIONGinaton could promote the mRNA expressions of the antiapoptotic gene bcl-2 and bcl-xL in myocardium of patients underwent CI'PB.

Cardiopulmonary Bypass ; methods ; Child ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression ; drug effects ; Ginkgo biloba ; chemistry ; Heart Defects, Congenital ; genetics ; surgery ; Humans ; Hypothermia, Induced ; Male ; Myocardium ; cytology ; metabolism ; Plant Leaves ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; bcl-X Protein ; genetics

OBJECTIVETo investigate the erythrocyte protective effects of Ginaton, a ginkgo biloba extract, in patients undergoing hypothermic cardiopulmonary bypass (CPB).

METHODSSixty patients, who suffered from rheumatic heart disease of ASA grade II-III and scheduled for mitral valve replacement with intravenous anesthesia, were randomly assigned to two groups equally, the Ginaton group and the control group. They were administered with Ginaton 1 mg/kg and saline respectively via intravenous dripping before open heart surgery before beginning CPB. Blood samples were taken from radial artery at different time points, i.e., before CPB (T1), nasopharyngeal temperature (30-31 degrees C) stabilized stage (T2), nasopharyngeal temperature restoration (36 degrees C) stage (T3), 30 min after CPB (T4) and 3 h after CPB (T5), for determination of malondialdehyde (MDA) and superoxide dismutase (SOD) levels in plasma and erythrocyte (P-MDA, E-MDA, P-SOD and E-SOD), as well as the Na+ -K+ -ATPase and Ca+ -Mg2+ -ATPase activities in erythrocytes.

RESULTSAs compared with those at T1, in the control group, P-MDA, E-MDA, and E-SOD at T2-T5 and E-SOD at T2 were higher, but E-SOD at T3-T5 were lower (P < 0.01); while in the Ginaton group P-MDA, E-MDA, and E-SOD at T3-T4 were higher (P < 0.05 or P < 0.01). As compared with those in the control group, the levels of P-MDA and E-MDA at T2-T5 were significantly lower, and E-SOD at T3-T5 were higher (P < 0.05 or P < 0.01). Activities of Na+ -K+ -ATPase and Ca+ -Mg2+ -ATPase significantly increased at T2 and gradually decreased after then in both groups (P < 0.05 or P < 0.01), but those at T2-T5 were significantly higher in Ginaton group than in control group (P < 0.05 or P < 0.01).

CONCLUSIONGinaton displays an erythrocyte protecting effect by way of alleviating the lipid peroxidation in erythrocytes' membrane.

Adult ; Cardiopulmonary Bypass ; methods ; Drugs, Chinese Herbal ; pharmacology ; Erythrocytes ; drug effects ; metabolism ; Female ; Ginkgo biloba ; Humans ; Hypothermia, Induced ; Lipid Peroxidation ; drug effects ; Male ; Middle Aged

OBJECTIVETo investigate the brain protective effects of Ginkgo biloba leaf extract (Ginaton) in patients who underwent hypothermic cardiopulmonary bypass (CPB).

METHODSSixty patients with rheumatic heart disease of ASA grade II-III, who were scheduled for mitral valve replacement with intravenous anaesthesia, were randomly assigned to two groups, the Ginaton group (30 patients) treated with Ginaton 1 mg/kg by intravenous dripping before open heart for CPB, and the control group (30 patients) with normal saline instead. Blood was synchronously collected from arteriae radialis and vena jugularis interna at 5 time points, namely, before CPB (T1), nasopharyngeal temperature (lowered to 30-31 degrees C) stabilized stage (T2), nasopharyngeal temperature restoration (36 degrees C) stage (T3), 30 min after CPB (T4) and 3 after CPB (Ts) for determining blood gas, lactate acid concentration, activity of superoxide dismutase (SOD) and malonaldehyde (MDA) content. And the oxygen content in artery (CaO2) and jugular vein (CjvO2), the difference of oxygen contents in arterial and jugular vein (Ca-jvO2), the cerebral oxygen extraction ratio (ERO2) as well as the arteriojugular lactate difference (ADVL) were calculated.

RESULTSAfter the beginning of CPB, as compared with those in the control group, in the Ginaton group, the reduction of Ca-jvO2 and ERO2 was significantly higher (P < 0.05 or P < 0.01) and the increase of lactate acid, ADVL and MDA were significantly lower, and with a remarkably higher SOD activity (P < 0.01).

CONCLUSIONGinaton could improve cerebral oxygen supply, promote SOD activity to inhibit production of free radicals in patients undergoing CPB, and thus shows an evident protective effect in the brain.

Adult ; Brain ; blood supply ; drug effects ; metabolism ; Cardiopulmonary Bypass ; methods ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Ginkgo biloba ; Heart Valve Diseases ; surgery ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Middle Aged ; Mitral Valve ; surgery ; Neuroprotective Agents ; therapeutic use ; Oxygen ; metabolism ; Phytotherapy ; Plant Extracts ; therapeutic use ; Plant Leaves ; chemistry ; Treatment Outcome

OBJECTIVETo investigate the effect and clinical value of ginkgo biloba extract (Ginaton) on the plasma vascular endothelial growth factor (VEGF) in patients during peri-operative period of cardiac surgery.

METHODSTwenty patients scheduled to receive cardiac operation were randomly assigned to 2 groups by a digital table. For the 10 patients in the control group, the cardiopulmonary bypass (CPB) was established in routine and received cold (4 degrees C) St. Thomas' cardioplegia perfusion (15 mL/kg) via aortic root after ascending aorta blocking, as for the 10 patients in the Ginaton group, the same was done but with 0.5 mg/kg of Ginaton added to the cardioplegia perfusion. Plasma VEGF contents were detected by ELISA at different time points, i.e., before and after anesthesia induction (T1, T2), after aorta intubation (T3), 0.5 h after aorta clamping (T4), 0.5 h after aorta declamping (T5), immediate after terminating the operation (T6), 6 h after operation (T7), and 24 h after operation (T8).

RESULTSIn the control group, VEGF level began to rise at T, and reached the peak at T7(P < 0.01), while in the Ginaton group, it reached the peak early at T, (P < 0.01), and began to drop at T (P < 0.01).

CONCLUSIONGinaton could induce the production of VEGF, which may be one of the mechanisms for its myocardial protection.

Cardiopulmonary Bypass ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Ginkgo biloba ; chemistry ; Heart Septal Defects, Atrial ; surgery ; Heart Septal Defects, Ventricular ; surgery ; Humans ; Male ; Perioperative Care ; Phytotherapy ; Plant Extracts ; therapeutic use ; Protective Agents ; therapeutic use ; Vascular Endothelial Growth Factor A ; blood

OBJECTIVETo investigate the mechanism underlying the effects of matrine in enhancing the cytotoxic sensitivity of CNE2/DDP cells highly expressing ATP-binding cassette superfamily G member 2 (ABCG(2)(High)) to allogenic natural killer (Allo-NK) cells.

METHODSABCG(2)(High) CNE2/DDP cells and Allo-NK cells were isolated by magnetic activated cell sorting (MACS). Flow cytometry was used to evaluate the purity of the isolated cells and the expression of NKG2D ligands on the target cells before and after incubation with matrine. The cytotoxic sensitivity of the treated and non-treated ABCG(2)(High) CNE2/DDP cells to Allo-NK cells was measured by LDH releasing assay.

RESULTSThe expression rate of ABCG2 was (91.40-/+2.32)% in ABCG(2)(High) CNE2/DDP cells. More than 90% of the isolated NK cells were identified to be CD3(-)CD16(+)CD56(+) cells. The expression rates of MICA, MICB, ULBP1, ULBP2, and ULBP3 on the target cells incubated with matrine increased from (2.92-/+0.33)%, (4.27-/+0.33)%, (5.80-/+0.62)%, (11.10-/+3.15)%, and (7.75-/+1.14)% to (11.30-/+0.89)%, (14.29-/+2.61)%, (12.56-/+1.06)%, (43.24-/+4.43)%, and (12.77-/+1.06)%, respectively. At the E: T ratio of 10:1 and 20:1, the cytotoxic sensitivity of ABCG(2)(High) cells to Allo-NK cells increased from (15.32-/+1.34)% and (27.26-/+6.81)% in un-treated cells to (28.53-/+1.37)% and (42.72-/+2.80)% in matrine-treated cells, respectively, showing significant differences in the cytotoxic sensitivity of the target cells in both groups produced by matrine treatment (F=29.05, P=0.000).

CONCLUSIONMatrine can up-regulate the expressions of NKG2D ligands (MICA/B and ULBP1-3) in ABCG(2)(High) nasopharyngeal carcinoma cells, which results in increased cytotoxic sensitivity of the cells to Allo-NK cells.

ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; metabolism ; Alkaloids ; pharmacology ; Carcinoma ; Cell Line, Tumor ; Flow Cytometry ; GPI-Linked Proteins ; metabolism ; Histocompatibility Antigens Class I ; metabolism ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Intracellular Signaling Peptides and Proteins ; metabolism ; Killer Cells, Natural ; drug effects ; metabolism ; Ligands ; NK Cell Lectin-Like Receptor Subfamily K ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Proteins ; metabolism ; Quinolizines ; pharmacology

OBJECTIVETo construct a phosphatidylinositol 4-kinase beta (PI4K-beta) mutant with the 325th to 373rd amino acid codons deleted, and try to develop a simple method for constructing middle fragment deletion mutant.

METHODSIn line with the mechanism of gene splicing by overlap extension(SOE), an additional PCR was used to get the PI4K-beta mutant in which the 325th to 373rd amino acid codons were deleted. Then the mutated gene was cloned into pCMV-Tag4A mammalian expression vector.

RESULTSA mutant with the 325th to 373rd amino acid codons deleted was constructed successfully.

CONCLUSIONThe improved SOE is a very effective and reliable method to construct middle fragment deletion mutant. It is worthy to be popularized.

1-Phosphatidylinositol 4-Kinase ; genetics ; Base Sequence ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; genetics ; Polymerase Chain Reaction ; methods ; Protein Engineering ; methods ; Recombinant Proteins ; genetics ; Sequence Deletion

OBJECTIVETo investigate the changes in the serum content of immunoreactive calcitonin (iCT) after burns or inhalation injury, and to explore its diagnostic significance.

METHODSTwenty-four dogs were randomized into 4 groups, i. e. A (n = 6, with moderate degree inhalation injury) , B ( n = 6, with severe inhalation injury), C (n = 6, with most severe inhalation injury) and D (n = 6, with severe burns) groups. The serum content of iCT and blood gas analysis before and after injury were determined at different time points. The degree of inhalation injury was determined with fibrobronchoscopic examination at 6 post-inhalation injury hour (PIH).

RESULTS(1) Fiber bronchoscopic examination showed that the degree of inhalation injury in each group was coincident with the anticipation. (2) The serum content of iCT in each group at 1 PIH was obviously higher than that before injury, and it was evidently higher in A, B and C groups than that in D group at 4 PIH. The peak value of iCT in group A at 24 PIH was (453+/-224) ng/L, and it increased gradually in B and C groups at 48 PIH. The serum content of iCT increased continually from 2 PIH on, and it reached (125+/-41) ng/L at 48 PIH. (3) Compared with PaO2 value before injury (109+/-8) mmHg, there was no obvious difference of the PaO, in A and D groups. PaO2 value in B and C group began to descend continually at 8 PIH (65+/-6) mmHg, and that in C group began to descend at 4 PIH (71+/-9) mmHg. PaCO2 value in C group began to increase at 24 PIH(52+/-11) mmHg when compared with that before injury(38+/-5 ) mmHg.

CONCLUSIONThe changes in the serum level of iCT within 8 PIH occurred much earlier than PaO2 and PaCO2, thus it has the same diagnostic significance as fibers bronchoscopic examination.

Animals ; Blood Gas Analysis ; Burns, Inhalation ; blood ; physiopathology ; Calcitonin ; blood ; Disease Models, Animal ; Dogs