In this paper, five different methods were carried out for DNA extraction directly from soil. The result shows that all five methods could generate DNA with more than 15 kb in size. They were subsequently used as templates for PCR amplification with success, using primers of the bacterial 16S rRNA gene and Shiva-1 gene encoding an antibacterial peptide. However, method 5 is more suitable for DNA extraction directly from a small amount of soil sample as it produced a good yield of DNA in high integrity with reliable reproducibility.

Objective To observe the clinical effect of Yiqi Yangyin Huoxue decoction(the function of decoction is to tonify Qi,nourish Yin and enhance blood circulation)on salivary gland response attribute to radiotherapy.Methods This study,carried between January 2005 and December 2005,focused on the effect of Chinese herbs on salivary gland response attribute to radiotherapy.In the treatment group,30 cases took Chinese herbs during the duration of radiotherapy,while in the control group 30 cases were given routine therapy.Results Both groups had finished the radiotherapy,however,in the control group,there were 5 cas- es with a break for 1～2 weeks.For the comparison of the salivary gland change in acute stage,there was no variance(x~2=2.387,P=0.122);the latency for the salivary gland change in treatment group was longer than that in control group(x~2=13.106,P=0.000).For the comparison of Karnofsky after radiotherapy,the KS was superior in the treatment group than that in control group(x~2=12.685,P=0.013);For the comparison of objective effect after radiotherapy,the remission rate in treatment group was 90 %,and it was 86.7 % in control group(x~2=0.638,P=0.727).Conclusion The decoction can remit the salivary gland response caused by radiotherapy in clinic,prolong the latency for acute radioactive response;release the pain of the pa- tients,increase the achievement ratio for radiotherapy,and improve the patients'living condition.To combine with radiotherapy,Chinese herbs is a good supplemental therapy for nasopharyngeal carcinoma

Fluorescence ; Host-Pathogen Interactions ; Luminescent Measurements ; methods ; Luminescent Proteins ; chemistry ; genetics ; metabolism ; Plant Diseases ; virology ; Plant Viruses ; chemistry ; genetics ; metabolism ; Plants ; chemistry ; genetics ; metabolism ; virology

OBJECTIVETo investigate a new method for correction of claw hand deformity after burns.

METHODSFrom May 2006 to Jul. 2010, 12 patients with claw hands deformities after burns were treated with skin grafts (11 hands) and skin flap (1 hand) with unsatisfactory results. Then elastic traction (skin traction or skeletal traction) were performed with individual functional brace.

RESULTSAll patients were followed up for 0.5 to 2 years. Elastic traction was effective in the correction of metacarpophalangeal joint deformity, buttonhole deformity, thumb-in-palm deformity, scar contracture, and palmar arch deformity.

CONCLUSIONSElastic traction is a simple and effective way for the correction of claw hand deformity after burns with less morbidity and stable results.

Adolescent ; Adult ; Burns ; complications ; Cicatrix ; surgery ; Female ; Follow-Up Studies ; Hand Deformities, Acquired ; etiology ; surgery ; Humans ; Male ; Middle Aged ; Skin Transplantation ; methods ; Surgical Flaps ; Traction ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo explore method of recombinant gene lentivirus containing LIM mineralization protein-1 (LMP-1) in transfecting bone marrow mesenchymal stem cells (BMSC), and to observe the effect of gene LMP-1 on proliferation effect and expression of BMSC.

METHODSSix clean SD rats aged 4 weeks were selected, bone marrow mesenchymal stem cells were extracted under sterile conditions and cultured to the third generation, then divided into three groups:control group (the third generation of BMSC), lentiviral vector transfection group (PGC-FU-GFP and Polybrene were injected into the third generation of BMSC) and recombinant gene transfection group (PGC-FU-LMP-1-GFP and Polybrene transfection were injected into the third generation of BMSC). After 48 hours' transfection, fluorescent expression were detected under immuno-fluorescence microscopy; lentiviral transfection efficiency were detected by flow cytometry; effect of lentiviral transfection on BMSC were evaluated by MTT; gene expression of transfected cells were determined by Western Blot.

RESULTS1) The third generation of BMSC was cultured successfully,and transfected with MOI:100. After 48 hours, green fluorescent expression were detected and transfection efficiency was 67% under immuno-fluorescence microscopy; 2) Compared to control group, there were no statistical differences between control group and other two groups; 3) Western blot teast showed that 72KDa specific band was observed in recombinant gene transfection group and its size was similar to LMP-1 fusion protein (50 kDa+28 kDa=78 kDa).

CONCLUSIONThere is no effect of recombinant gene lentivirus containing LIM on BMSC, and can effectively influence the expression of LMP-1.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Animals ; Cell Proliferation ; Cells, Cultured ; Cytoskeletal Proteins ; genetics ; metabolism ; Female ; Genetic Therapy ; Genetic Vectors ; genetics ; metabolism ; Humans ; LIM Domain Proteins ; genetics ; metabolism ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; virology ; Osteoporosis ; genetics ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley

Objective:To investigate the underlying mechanism of histone deacetylase (HDAC) inhibitor FK228-in- duced apoptosis of the prostate cancer cell line DU145.Methods:The inhibitory effect of FK228 on DU145 cell growth and its cytotoxicity were determined by MTT assay;cell cycle arrest was detected by flow cytometry assay;morphological change was observed by Giemsa staining;and defined kinase protein levels were determined by Western blot analysis.Re- suits:FK228 obviously inhibited DU145 cells growth,arrested cell cycle at G_0/G_1 phase,induced cells morphological changes and degraded several kinase proteins,including EGFR,Her2,Raf-1,Src,Cdk4 and IAP member Survivin.The degradation of these kinases blocked Raf-Mek-Erk and PI3K/Akt survival signal pathways,inducing apoptosis.Condu- sion:FK228 may induce DU145 cell apoptosis through depletion of multiple kinase proteins and blockade of survival sig- nal pathways of DU145 cells.

Objective: To study the factors and preventive method of post-operative complications of spinal implant operation. Methods: The post-operative complication of 451 cases who had undergone spinal implant operation were analysed retrospectively. The relative factors of complications were analyzed. Results: Infection, thrombosis of deep vein and failed fusion were the most common complications. Among the factors, the age, operation time and blood lose during operation were closely related with the complication. Using antibiotic was effective to reduce the infection. Conclusion: It is effective to reduce the post-operative complication of spinal implant operation by proper pre-operative evaluation. Using antibiotic to control other diseases is also effective.

AIM:To investigate the mechanism of quercetin improving rat coronary artery myogenic response under high glucose ( HG) by measuring muscle tension of coronary arterial ring and recording voltage -gated K +channel ( Kv) current of coronary artery smooth muscle cells by whole cell patch clamp .METHODS:The coronary rings from the normal SD rats were acutely isolated , and then divided into 6 groups:(1) control group;(2) HG group;(3) HG+low dose (3 μmol/L) of quercetin group;(4) HG+moderate dose (10 μmol/L) of quercetin group; (5) HG+high dose (30 μmol/L) of quercetin group;(6) HG+C6303 (PKC inhibitor) +high dose of quercetin group.Determinations of coronary artery response to vasoconstrictor (60 mmol/L KCl or 0.1 mmol/L U46619) or vasodilator (Ach at 10 -9 ~10 -5 mol/L) were performed, and the percentage of coronary ring tension was calculated using the contraction as 100%caused by 60 mmol/L KCl.The rat coronary artery smooth muscle cells were acutely isolated for recording the Kv current using whole cell patch clamp .RESULTS:Compared with control group , the contraction amplitudes to 60 mmol/L KCl or 0.1 mmol/L U46619 were significantly increased under HG incubation .Quercetin intervention concentration-dependently re-duced the coronary artery contraction amplitude .Incubation of PKC specific inhibitor C 6303 attenuated the effect of querce-tin.Compared with control group , the diastolic amplitude to Ach decreased significantly in HG group , and quercetin inter-vention concentration-dependently increased the coronary artery diastolic amplitude .Incubation of PKC specific inhibitor C6303 attenuated the effect of quercetin .Compared with control group , HG incubation inhibited Kv current of coronary ar-tery vascular smooth muscle cells significantly , and quercetin intervention attenuated the inhibitory effect of HG on Kv cur-rent intensity .Incubation of PKC specific inhibitor C 6303 attenuated the effect of quercetin .CONCLUSION: Quercetin has a protective effect on myogenic response of coronary artery under HG and the effects is related to the increase in Kv cur -rent and the activation of PKC in vascular smooth muscle cells .

OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute-olin-7-O-β-D-glucuronide (LGU) against focalcerebral ischemic injury. METHODS The focal cerebral ischemic injury model was established by middle cerebral artery occlusion (MCAO). Male Sprague Dawley rats were randomly divided into sham group,model group(MCAO),LGU group(0.24,0.72 and 2.16 mg·kg-1)and positive control group(Edaravone at 5 mg·kg-1).LGU was injected intravenously 30 min after MCAO.Neurological severity score,infarct volume and brain water content were detected 24 h after MCAO and the levels of Na+-K+ATPase,Ca2+ATPase,TNF-α and IL-1β were detected to explore the possible mechanisms.For the therapeutic time window test,LGU(0.72 mg·kg-1)was injected intrave-nously 0.5, 2, 4, 6, 8, 10 and 12 h respectively after MCAO. To evaluate motion behavior, LGU were injected intravenously 30 min after MCAO and once per day during detection period. The changes of motor coordination were detected by rotating rod method and grip strength analysis, and the changes of gaits were detected using DigiGait Imaging System. RESULTS LGU improved the neurological severity score, infarct volume ratio and brain water content. The therapeutic time window of LGU for cerebral infarction and brain edema was at least 6 h and for neurological dysfunction was 12 h.LGU also prolonged the latency on rotarod, increased the forelimb tension and improved 8 gait parameters, including stance duration,stride length,stance width,paw area,paw area variability,gait symmetry,ataxia coefficient and tau propulsion.Furthermore,LGU increased Na+-K+-ATPase and Ca2+-ATPase levels in the cortex and hippocampus in the ischemic side,reduced the levels of TNF-α and IL-1β in the serum. CONCLUSION LGU has a significant neuroprotective effect against cerebral ischemic injury via improving energy metabolism and reducing inflammation.

OBJECTIVETo investigate the related factors and pathogens of ventilator-associated pneumonia (VAP) after heart surgery.

METHODSVAP was diagnosed in 105 patients out of 1688 cases (6.2%) who underwent heart surgery in our department between January 2004 and January 2011. Clinical data, pathogens and treatments were analyzed.

RESULTSIncidence of VAP was 6.2% (105/1688), and 53.0% (105/198) in patients who required more than 48 hours of mechanical ventilation. One hundred and ninety-eight pathogen strains were isolated by bacterial culture, in which Gram negative bacteria accounted for 69.2% (137/198), Gram positive bacteria 27.8% (55/198), and fungi for 3.0% (6/198). Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, and Staphylococcus aureus were the main pathogens of VAP. The independent risk factors for VAP were: age > 70 years, emergent surgery, perioperative transfusions, reintubation and days of mechanical ventilation (all P < 0.01). Median length of stay in the ICU for patients who developed VAP or not was (24.7 ± 4.5) days versus (3.2 ± 1.5) days, respectively (P < 0.05) and in-hospital mortality was 25.7% (27/105) versus 2.9% (46/1583) respectively (P < 0.05).

CONCLUSIONSPatients undergoing heart surgery have a high frequency of developing VAP, especially in patients that require more than 48 hours of mechanical ventilation. VAP is associated with high in-hospital mortality. Age > 70 years, emergent surgery, perioperative transfusions, reintubation and prolonged mechanical ventilation use are independent risk factors for VAP in patients following cardiac surgery. Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, and Staphylococcus aureus are the main pathogens of VAP.

Acinetobacter baumannii ; Adult ; Aged ; Cardiac Surgical Procedures ; adverse effects ; Female ; Humans ; Klebsiella pneumoniae ; Male ; Middle Aged ; Pneumonia, Bacterial ; microbiology ; Pneumonia, Ventilator-Associated ; microbiology ; Pseudomonas aeruginosa ; Retrospective Studies ; Risk Factors