Objective To establish the ELISA kit of monoclonal antibodies to Clostridium difficile toxin A.Methods A sandwich ELISA was used.Flat-bottomed 96 well polystyrene microtitre plates were coated with 100 ?l of purified rabbit monospecific antitoxin(8 ?g/ml, capturing antibody) in carbonate buffer(pH9.6) and incubated overnight at 4℃, the plates were washed once in PBS containing 0.05% Tween-20, pH7.4 (PBST). After 200 ?l of 10% BSA in PBS-T was added to the wells and incubated at 37℃ for 2h, washed 5 times in PBS-T with 3 min incubation at room temperature between each wash, 100 ?l of C. difficile toxin A or test samples in PBS-T were added to each well and incubated for 1h at 37℃, washed 5 times. Then 100 ?l of 1:1000 diluted monoclonal antibodies IgG-Horseradish peroxidase conjugate(detecting antibody) was added for 1h at 37℃, wells were washed five times with PBS-T, and 0.1ml of tetramethylbenzidines substrate was added to each well. After 15 min at 37℃ in dark, the reactions were stopped by the addition of 1 drop of 2M sulfuric acid and the A450 was measured.Results The tested specimens included culture filtrates of 2 strains of toxigenic C. difficile, 2 non-toxigenic strains of C. difficile, 26 strains of E. coli, 2 strains of S. dysenteriae, 1 strains of Bifidobacterium, 5 strains of V. cholera, 2 strain of S. typhi, 7 strains of C. botulinum, 1 strain of toxigenic C. sordllii, and 1 strain of C. butyricum. The ELISA demonstrated high specificity and good sensitivity, it detected amounts of toxin A as low as 0.1ng/ml.Conclusion An ELISA kits with high specificity and good sensitivity for the rapid detection of C. difficile toxin A was presented. It will be a beneficial tool to clinical detection of Clostridium difficile toxin A.

OBJECTIVETo investigate clinical effects of damage contral resuscitation (DCR) in rescue multiple fracture with hemorrhagic shock.

METHODSFrom January 2009 to May 2013, clinical data of 24 patients suffered from multiple fracture with hemorrhagic shock were retrospectively reviewed. Among them, 18 cases were male and 6 cases were female, aged from 21 to 48 years old with an average of (32.5 +/- 4.5). Small capacity of balanced salt solution were used to maintain systolic, pressure (80-90) mmHg before operation. After control of bleeding with operation, sufficient amount of liquid were rapidly use, and plasma were supplied according to proportion of plasma and red cell suspension (1 U:2 U-1 U:1 U). After remedy of shock, fluid infusion were sustained negative balance slightly and keep acid-base equilibrium of electrolyte. Dosage of balanced salt solution and blood transfusion before remedy shock were recorded, removal time of lactic acid, coagulation function, incidence and case fatality of DIC were observed.

RESULTSFour patients were died after 6-18 h into hospital (2 cases died for acute respiratory distress syndrome and 2 cases for irreversible shock). Twenty patients with shock were corrected at 2-6 h after injury. Dosage of balanced salt solution was(4,259 +/- 268) ml,red cell suspension was (14 +/- 2) U, fresh frozen plasma was (800-1,600) ml (FFP: 1 U = 100 ml) averaged (900 + 300) ml, blood platelet was 4-6 U. Coagulation function and electrolyte were normal. Lactic acid was less than 2 mmol/L within 24 h,the success rate of recovery was 83.3% (20/24).

CONCLUSIONPerforming DCR can obvious improve success rate of remedy serious fracture combined with uncontrolled hemorrhagic shock. Supplementing FFP when correcting coagulation function should be carry out promptly in fluid resuscitation.

Adult ; Blood Transfusion ; Female ; Humans ; Male ; Middle Aged ; Multiple Trauma ; therapy ; Resuscitation ; Retrospective Studies ; Shock, Hemorrhagic ; therapy ; Young Adult

OBJECTIVETo discuss the clinical effects of damage control resuscitation (DCR) in treating severe multiple trauma in the pelvic fractures.

METHODSFrom March 2009 to September 2013, a restrospective analysis was conducted on the clinical data of 28 patients with multiple trauma in the pelvic fractures, including 19 males and 9 females, ranging in age from 21 to 51 years old with an average of 32.5 years old. According to Tile classification of pelvic fractures, 16 cases were type B and 12 cases were type C. Injury severity score (ISS) averaged 32.0±3.4. All cases were treated with damage contral orthopaedics and DCR strategy, namely that used immediately with small capacity of balanced salt solution on admission so as to maintain the blood pressure between 80 to 90 mmHg. At the same time, emergency phase I simple debridement plus external fixator or bundled with fixed pelvic was done, and the chest or abdomen combined injury was treated at first. And limbs fracture was temporary dealing with bone traction or plaster external fixation, etc. After bleeding was controlled by operation, fluid resuscitation was done as fast as possible and the plasma was transfused early according to the proportion of plasma and red cell suspension (2 U:1 U-3 U:2 U). Dosage of balanced salt solution and blood before remedy shock was recorded, and the removal of time of lactic acid and coagulation were observed.

RESULTSFive cases were died after 4 to 15 hours into hospital (3 cases died for severe trauma-hemorrhagic shock and 2 cases for acute respiratory distress syndrome). Twenty-three cases were remedy shock at 1.6 to 4.3 hours after injury with an average of 2.4 hours. Period I operation duration was (78.2±10.3) minutes. Dosage of balanced salt solution was (3,798±340) ml and red cell suspension was 14 to 18 U, fresh frozen plasma (FFP) was (1,267±58) ml (1U FFP=100 ml), blood platelet was 8 to 12 U for 9 patients. The removal time of lactic acid and PT-APTT was (11.4±2.1) hours and (4.3±0.8) hours. Measures were taken to correct electrolyte and acid-base imbalance was normal. The success rat of recovery was 82.2% (23/28).

CONCLUSIONDamage control resuscitation (DCR) integrates the main links such as damage control operation and allowable low blood pressure (or limited liquid resuscitation) and hemostatic control resuscitation, was early effective treatment measures for the patients with severe multiple trauma in the pelvic fractures.

Adult ; Female ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Multiple Trauma ; surgery ; Pelvic Bones ; injuries ; surgery ; Resuscitation ; Retrospective Studies ; Treatment Outcome ; Young Adult

Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Ischemic Attack, Transient ; drug therapy ; Male ; Middle Aged ; Phytotherapy

Objective: To study the benzo(a)pyrene (B[a]P)-induced mRNA expression of aromatic hydrocarbon receptor (AHR) and cytochrome P4501A1 (CYP1A1) genes in rat liver. Methods: Rats were injected intraperitoneally with 5, 10 and 15 mg/kg of B[a]P. The total RNAs were extracted from rat livers by RNA purification kit, and the mRNA expression of AHR and CYP1A1 genes was determined by reverse transcription polymerase chain reaction (RT-PCR). β-actin was used as the internal control. The mRNA expression of both AHR and CYP1A1 genes was measured at indicated time points (24, 48 and 72 h) after B[a]P treatment at three different concentrations (5, 10 and 15 mg/kg). Results: The mRNA expression of AHR gene increased in a time-dependent manner at the concentration of 10 mg/kg but not at 5 and 15 mg/kg of B[a]P. The mRNA expression of CYP1A1 gene differed significantly at 48 h and 24 h in rat livers treated with 10 and 15 mg/kg dosage of B[a]P. The mRNA expression of AHR and CYP1A1 genes increased with B[a]P treatment in a concentration-dependent manner. The time-dependent increase in mRNA expression was shown by AHR but not by CYP1A1 gene with B[a]P (10 mg/kg) treatment. Conclusion: This study demonstrates that toxic B[a]P increases the mRNA expression of both AHR and CYP1A1 genes in vivo, suggesting that B[a]P may play a role in cancer genesis by this way.

Objective: To estimate the relative risk for lung cancer associated with genetic polymorphism of T6235C mutation in 3′ non-coding region (MspI) of cytochrome P450 1A1 (CYP1A1) and glutathione S-transferase M1(GSTM1) in the Mongolian population in Inner Mongolian Region of China. Methods: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and multiplex PCR methods were used to analyze blood samples obtained from 263 case subjects and 263 control subjects to determine their genotypes for CYP1A1 and GSTM1. Control subjects were matched with case subjects by ethnic background, age and gender. Results: The frequencies of the variant CYP1A1 genotypes (CYP1A1C) and GSTM1-null in lung cancer groups were higher than those in control groups (38.4% vs. 28.5% and 57.8% vs. 48.0%). The individuals who carried with CYP1A1C genotype had a significantly higher risk of lung cancer (OR=1.56, 95% CI=1.08 to 2.25, P=0.016) than those who carried with non-variation CYP1A1 genotype. The ones who carried with GSTM1-null genotype also had a significantly higher risk of lung cancer (OR=1.49, 95% CI=1.06 to 2.10, P=0.023) than those who carried with GSTM1-present genotype. When combination of polymorphisms of CYP1A1 and GSTM1 genotypes was analyzed, the risk of lung cancer for combination of CYP1A1C and GSTM1-null genotypes was increased significantly (OR=2.084, 95% CI=1.27 to 3.42, P=0.003). Susceptibility to lung cancer was related to smoking (OR=2.10, 95% CI=1.48 to 2.98, P=0.000). Considering smoking status, the risk of lung cancer for combination of smoking and CYP1A1C genotype was remarkably increased (OR=2.76, 95% CI=1.74 to 4.37, P=0.000). It was the same case with combination of smoking and GSTM1-null genotype (OR=4.38, 95% CI=2.35 to 8.15, P=0.000). Conclusion: The polymorphisms of CYP1A1C genotype and GSTM1-null are the risk factors of lung cancer in the Mongolian population in Inner Mongolia Region of China. Smoking is also related to susceptibility to lung cancer. There may be a synergetic interaction between CYP1A1C and GSTM1-null in the elevated susceptibility of lung cancer. Smoking may have a synergetic interaction with CYP1A1C and GSTM1-null in the elevated susceptibility of lung cancer.

Objective: To study the benzo(a)pyrene (B[a]P)-induced mRNA expression of aromatic hydrocarbon receptor (AHR) and cytochrome P4501A1 (CYP1A1) genes in rat liver. Methods: Rats were injected intraperitoneally with 5, 10 and 15 mg/kg of B[a]P. The total RNAs were extracted from rat livers by RNA purification kit, and the mRNA expression of AHR and CYP1A1 genes was determined by reverse transcription polymerase chain reaction (RT-PCR). β-actin was used as the internal control. The mRNA expression of both AHR and CYP1A1 genes was measured at indicated time points (24, 48 and 72 h) after B[a]P treatment at three different concentrations (5, 10 and 15 mg/kg). Results: The mRNA expression of AHR gene increased in a time-dependent manner at the concentration of 10 mg/kg but not at 5 and 15 mg/kg of B[a]P. The mRNA expression of CYP1A1 gene differed significantly at 48 h and 24 h in rat livers treated with 10 and 15 mg/kg dosage of B[a]P. The mRNA expression of AHR and CYP1A1 genes increased with B[a]P treatment in a concentration-dependent manner. The time-dependent increase in mRNA expression was shown by AHR but not by CYP1A1 gene with B[a]P (10 mg/kg) treatment. Conclusion: This study demonstrates that toxic B[a]P increases the mRNA expression of both AHR and CYP1A1 genes in vivo, suggesting that B[a]P may play a role in cancer genesis by this way.

Objective: To estimate the relative risk for lung cancer associated with genetic polymorphism of T6235C mutation in 3′ non-coding region (MspI) of cytochrome P450 1A1 (CYP1A1) and glutathione S-transferase M1(GSTM1) in the Mongolian population in Inner Mongolian Region of China. Methods: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and multiplex PCR methods were used to analyze blood samples obtained from 263 case subjects and 263 control subjects to determine their genotypes for CYP1A1 and GSTM1. Control subjects were matched with case subjects by ethnic background, age and gender. Results: The frequencies of the variant CYP1A1 genotypes (CYP1A1C) and GSTM1-null in lung cancer groups were higher than those in control groups (38.4% vs. 28.5% and 57.8% vs. 48.0%). The individuals who carried with CYP1A1C genotype had a significantly higher risk of lung cancer (OR=1.56, 95% CI=1.08 to 2.25, P=0.016) than those who carried with non-variation CYP1A1 genotype. The ones who carried with GSTM1-null genotype also had a significantly higher risk of lung cancer (OR=1.49, 95% CI=1.06 to 2.10, P=0.023) than those who carried with GSTM1-present genotype. When combination of polymorphisms of CYP1A1 and GSTM1 genotypes was analyzed, the risk of lung cancer for combination of CYP1A1C and GSTM1-null genotypes was increased significantly (OR=2.084, 95% CI=1.27 to 3.42, P=0.003). Susceptibility to lung cancer was related to smoking (OR=2.10, 95% CI=1.48 to 2.98, P=0.000). Considering smoking status, the risk of lung cancer for combination of smoking and CYP1A1C genotype was remarkably increased (OR=2.76, 95% CI=1.74 to 4.37, P=0.000). It was the same case with combination of smoking and GSTM1-null genotype (OR=4.38, 95% CI=2.35 to 8.15, P=0.000). Conclusion: The polymorphisms of CYP1A1C genotype and GSTM1-null are the risk factors of lung cancer in the Mongolian population in Inner Mongolia Region of China. Smoking is also related to susceptibility to lung cancer. There may be a synergetic interaction between CYP1A1C and GSTM1-null in the elevated susceptibility of lung cancer. Smoking may have a synergetic interaction with CYP1A1C and GSTM1-null in the elevated susceptibility of lung cancer.

Objective To study the curative effect of major aphthous ulcer using povidone-iodine,H_2O_2 asso- ciated with levamisole liniment.Methods The RAU patients were divided into 2 groups randomly.The patients of curative group were instructed to use povidone-iodine,H_2O_2 and levamisole liniment;The control group were in- structed to take metronidazole,antibiotics,compound vitamin B and vitamin C.Then the patients were observed peri- odically.Results The success rate of curative group was 85 % and the control group was 55 %.The difference had statistical significance(P＜0.01).Conclusion Treatment of major aphthous ulcer using povidone-iodine,H_2O_2 and levamisole liniment is effective.

BACKGROUND:Bone marrow mesenchymal stem cells have beneficial effects on the treatment of various diseases. Mitochondria transfer is newly proposed and its specific mechanisms of action and control factors remain unclear.
OBJECTIVE:To review the studies about stem cells and mitochondrial transfer, then to discuss its value on clinical.
METHODS:The PubMed, VIP and Wanfang databases were searched for related articles concerning stem cells and mitochondrial transfer. Key words were“stem cell, embryonic stem cell, progenitor cell, mitochondria”in Chinese and“Stem cell[s], Mother cell[s], Progenitor cell[s], Colony-Forming Unit[s], Colony Forming Unit[s], Mitochondria[l] transfer”in English. Thirteen articles were found initial y, and using the citation tracking method, final y 42 articles were determined. RESULTS AND CONCLUSION:Studies have shown that mitochondrial transfer from bone marrow mesenchymal stem cells is associated with rescue of aerobic respiration and restoration of mitochondrial function in the injured somatic cells. Bone marrow mesenchymal stem cells and recipient cells form tunneling nanotubes for mitochondrial transfer. Movement of mitochondria between cells is regulated and directed by Miro1. The successful transfer of mitochondria may be accompanied with the clearance of damaged mitochondria. The rescue of mitochondrial function in early stages may provide a valuable therapeutic strategy for various diseases include acute lung injury.