Objective To study the significance of detecting the mutations of K-ras gene in faecal for diagnosis colorectal carcinoma.Methods 36 cases suffering from colorectal carcinoma,24 cases from colorectal benign diseases and 24 cases from healthy subjects,all cases had been diagnosed by enteroscope. K-ras gene mutation at the codon 12 was detected by PCR-RFLP,DNA was taken from faecal in modified method.Results K-ras mutation rate in colorectal carcinoma was 77.8 %,colorectal benign diseases was 25.0 %,healthy cases was 8.3 %.Conclusion Detecting the mutations of K-ras gene in faecalmay improve the rate of diagnosing colorectal carcinoma.

Objective To evaluate the effectiveness of urgent surgical correction for infra-cardiac total anomalous pulmonary venous drainage (TAPVD) in infants and children. Methods From July 2000 to April 2009, seventeen patients with infra-cardiac type of total anomalous pulmonary venous connection received surgical correction in our department. The age of patients ranged from 15 days to 3.7 months at admission, 9 patients ( 52.9 % ) were younger than 1 month of age. The mean body weight was (4.10 ±0.95) kg and was less than 5 kg in 15 patients (88.2%). Deep hypothermic circulatory arrest (DHCA) without cerebral perfusion was used in 13 patients (76%), and continuous hypothermic low-flow perfusion was used in4 (24%). Incisions on the left atrium and pulmonary veins were sutured without tension, and the width of the anastomosis was 2.5 to 4.0 cm. Results Urgent operations were performed in all patients and no early death occurred during surgery.Mean bypass time was 121.9 minutes ( ranging from 70 to 277 minutes) and mean aortic cross-clamp time was 44 minutes ( ranging from 30 to 74 minutes). 6 neonates had delayed wound closure after surgery and 4 had obvious pulmonary edema. Inhalation of nitric oxide was given to 5 cases for the management of significant postoperative pulmonary hypertension, while peritoneal dialysis was performed in 2 cases with temporary renal failure. Transient atrioventricular block of grade 3 occurred in 2patients and acute renal failure occurred in 2 cases. All patients were uneventfully discharged. The mean ventilation time was 45 h (ranging from 12 to 430 h). Mean duration in ICUs and hospitals were 15 days ( ranging from 4 to 40 days) and 22. 5days ( ranging from 10 to 42 days), respectively. During the period of follow up, one patient presented with pulmonary venous obstruction and recurrent pulmonary infection with increased flow speed at the site of anastomosis, pulmonary hypertension and tricuspid regurgitation on echocardiogram. No improvement was observed in the patient spite of medical treatment and he was lost during follow up one year after operation. 16 patients received postoperative follow up for 1 month to 9 years, mean (51.8 ± 35.0) months. Echocardiography, chest radiography, and ECG were performed during this period. As a result, most children had good cardiac function, with sinus rhythm on ECG and apparently reduced pulmonary congestion on radiography.No obstructive pulmonary venous return was observed on echocardiography. Most patients had good cardiac functions. Conclusion Corrective operation for infra-cardiac TAPVC on urgent basis may provide favorite outcomes. The prognosis is associated with the size of anastomosis between the pulmonary vein and left atrium, as large anastomosis may prevent the obstruction of pulmonary venous retum.

To study the effect of cholesterol and 25-OH-cholesterol on cholesterol metabolism in HepG2 cells and the effect of coptisine (Cop) extracted from Coptidis Rhizoma (CR) in reducing and regulating cholesterol. In this study, TC, TG, LDL-c and HDL-c were measured by biochemical analysis; mRNA and protein expressions of LDLR, HMGCR and CYP7A1 were detected by qRT-PCR and Western blot. According to the results, cholesterol and 25-OH-cholesterol inducing could decrease in mRNA and protein expressions of LDLR and CYP7A1, so as to increase TC and LDL-c contents. However, Cop could up-regulate mRNA and protein expressions of LDLR and CYP7A1 and down-regulate that of HMGCR, so as to reduce TC and LDL-c levels. These findings suggested that Cop has potential pharmacological activity for reducing cholesterol, and may reduce cholesterol by regulating mRNA and protein expressions of key genes involved in cholesterol metabolism, such as LDLR, CYP7A1 and HMGCR. This study laid a firm theoretical foundation for developing new natural drugs with the cholesterol-lowering activity.
Berberine ; analogs & derivatives ; pharmacology ; Cholesterol ; metabolism ; Cholesterol 7-alpha-Hydroxylase ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Enzymologic ; drug effects ; Hep G2 Cells ; Humans ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Receptors, LDL ; genetics ; metabolism ; Triglycerides ; metabolism

Objective To investigate the importance of plasmid-mediated quinolone resistance in the development of quinolone resistance in clinical isolates of gram-negative bacteria.Methods A total of 541 consecutive clinical isolates of gram-negative ba- cilli resistant or intermediate to ciprofloxacin were screened for the qnrA gene by PCR.Conjugation experiments were carried out with azide-resistant E.coli J53 as a recipient.The aac(6')-Ib-cr gene was detected.The mutations in the quinolone-resist- ance-determining region (QRDR) of the gyrA and parC genes were identified in qnrA positive strains.Results qnrA was identi- fied in 7 of the 541 strains.Among the qnrA positive strains,5 were Enterobacter cloacae.No qnrA was detected in nonfer- menters.Quinolone resistance was transferred in 4 of 7 qnrA positive strains.Transconjugants had 12-to 125-fold increases in MIC of ciprofloxacin relative to that of the recipient.Seven strains contained qnrA with a nucleotide sequence identical to that originally reported.Two transconjugants with higher ciprofloxacin MICs contained aac(6')-Ib-cr gene.Mutations occurred in the QRDR of the gyrA and parC genes in 5 PCR-positive clinical strains.Conclusions Transferable plasmid-mediated quinolone resistance associated with qnrA is highly prevalent in clinical strains of Enterobacter spp.aac(6')-Ib-cr gene and mutations in the quinolone targets may co-exist with qnrA,which may contribute to the further increase of resistance to quinolones.

Objective To evaluate the early clinical efficaoy of hybrid procedure for infants less than six months old with ventricular septal defect and coarctation of aorta.Methods From January 2010 to July 2011,20 patients with ventricular septal defect and coarctation of aorta received hybrid procedure in our center.The body weight was (4.5 ± 1.6) kg ( ranged from 1.9 kg to 6.5 kg) and the age was ( 56 ± 45 ) days ( ranged from 18 days to 6 months).The pressure gradient of the coarctation of the aorta ranged from 30 mm Hg to 56 mm Hg,5 patients of them were diagnosed as hypoplasty of aortic arch.The size of the ventricular septal defect ranged from 8 mm to 16 mm.Results The mortality was zero in all the 20 cases during the surgery,and the mobidity was 20％ (4/20).The complications were pneumonia in 2 cases,infective endocarditis in 1 case and pneumothorax in 1 case.The diameter of coarctation of the aorta ranged from 1.5 mm to 3.4 mm,and the size of the balloon ranged from 4 mm to 12 mm.The pressure gradient of the coarctation of the aorta decreased to 0 to 27 mm Hg.The bypass time ranged from 40 minutes to 87 minutes,and the crossclamp time of the aorta ranged from 20 minutes to 41minutes.The atrial septal defects were repaired and the patent ductuses were ligated during the surgery without leaving the sternum open.The total operation time was (4.0 ± 0.7 ) hours ( ranging from 3.0 hours to 5.2 hours).The mean ventilation time was (2.2 ± 1.4) days and mean ICU stay time was (5 ± 3 ) days.All the patients were followed up for ( 10.0 ± 3.6) months without aneurysm in arch and obstruction in airway.The residual obstructive pressurc gradicnt in the aortic arch ranged from 12 mm Hg to 35 mm Hg and 2 patients received reintervention.One patient received re-balloon dilation and the other received surgery.The cardiac function reached NYHA Ⅰ - Ⅱ in all eases.Conclusion The early outcome of the hybrid procedure (balloon dilation of the coartation of the aorta and surgical repair of ventricular septal defect) for infants with ventricular septal defect and coarctation of aorta was satisfying,which could avoid from circulatory arrest.It is a relatively safe procedure which could be the optional method for one-stage surgical repair.

Objective To explore the characteristics of ischemia-reperfusion induced infant lung damage and the potential mechanisms of the injuried.Methods Both infant (15-21 days old) and adult (5-6 months old) rabbits were subjected to either ischemia-reperfusion or sham operation.Ischemia-reperfusion was induced by clamping the right pulmonary hilum for 1 hour and then removal of the clamp for 4 hours under anesthesia.The lung tissue were sampled for histological examination by light and electron microcopies and for biological evaluation of mitochondrial alterations.Production and expression of free radical species-hydroxyl radical (ROS-HR),malondialdehyde (MDA),superoxide dismutase (SOD),glutathione peroxidase (GSH-PX),myeloid differentiation factor-88 (MyD-88),and nuclear factor-κB (NF-κB) in the lung tissue were also examined.In addition,circulating levels of interleukin-β and tumor necrosis factor-α were measured during the ischemia-reperfusion process.Results In comparison to adult lungs,the infant lungs had more increased neutrophil infiltration,edema,swelled alveolar epithelial and endothelial cells,and severer mitochondrial impairment reflected by damage of the inner membrane as well as decrease in the membrane potential after ischemia-reperfusion.The lungs in infant animals subjected to sham operation displayed higher levels of ROS-HR and MDA and lower levels of SOD and GSH-PX than those in adult controls.The lungs in infants with ischemia-reperfusion were found to further produce more ROS-HR,and MDA,and less SOD and GSH-PX than the ischemia-reperfused adult lungs.Moreover,the circulating levels of interleukin-1β and tumor necrosis factor-α were elevated during the period of ischemia-reperfusion,particularly in the infant animals,which appeared to be associated with the expression of MyD-88 and NF-κB in the lungs.Conclusion Lung ischemia-reperfusion causes more severe lung damage in infants than in adults,probably due to combination of low antioxidant capacity and overproduction of ROS in infants.

Objective To establish a novel experimental model of New Zealand rabbit to assess the biocompatibility of 0.1 mm polytetrafluoroethylene (PTFE),a novel material of pulmonary valve.Methods Forty-two adult New Zealand rabbits about 3 kg were selected to give 35 mg/kg ketamine and 0.25 mg/kg dexmedetomidine intramuscularly for the anesthesia.The chest was open to expose the upper segment.The surface of right ventricular outflow tract (RVOT) was exposed after the pericardium was opened partially.The valve material (0.1 mm PTFE) was inserted into the right ventricle via the central mini-incision.Then the skin was closed and the rabbits were recoved with the oxygen inhalation.Results Forty-two adult New Zealand rabbits accepted the operation.Six rabbits died during the early period due to the inappropriate anesthesia drug (n =2),pneumothorax (n =1) and thrombosis (n =3),which occurred the 5th,7th and 8th postoperative day.The anticoagulation treatment was adjusted to solve the thrombosis problem.Thirty-six rabbits survived for several months with weight increase until the experiment was finished.Conclusions The experimental model of New Zealand rabbit is appropriate for the biocompatible assessment of 0.1 mm PTFE.The advantage is to avoid intubation and cardiopulmonary bypass (CPB) and to decrease the pneumothorax.

Aim The effect of tetrandrine on TGF-?1 mRNA expression in glomerulosclerosis rat was observed. Methods The rats were randomly divided into four groups, such as the normal control group (sham operative rat), glomerulosclerosis model group,tetrandrine group and amlodipine group. The expression of TGF-?1 mRNA was analyzed by Northern blot hybridization. Results The expressions of TGF-?1 mRNA in two treating groups were much lower than untreated model group. There were no difference between these two treating groups. Conclusion Tetrandrine can decrease the expression of TGF-?1 mRNA in glomerulosclerosis rat induced by unilateral renctomy plus adriamycin.

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment

OBJECTIVETo investigate the effect of Astragalus membranaceus (AM) on endothelial-dependent (EDV) and non- dependent (EIV) vascular relaxation in ex vivo thoracic aortic rings of obese rats.

METHODSFifteen SD rats were randomized into 3 equal groups, namely the control group fed with normal chow, obese group with high-fat chow, and AM intervention group fed with high-fat chow and daily AM gavage. The rats were sacrificed after 6 weeks of feeding, and the aortic rings were dissected and cut into 3-mm rings. The response to acethylcholine (Ach) and sodium nitroprusside (SNP) were examined in organ bath. In ex vivo study, the aortic rings obtained from the control group and obese group were incubated with AM or vehicle for 3 h in organ bath before testing the EDV and EIV. The body weight and weight of the visceral fat in each group were recorded.

RESULTSThe weight of visceral fat was greater in the obese group than in the control group, and a 6-week AM treatment significantly reduced the fat tissue due to high-fat diet. The maximum EDV value was (87.0 - or + 3.5)% in the control group, (54.8 - or + 7.8)% in the obese group, and (69.8 - or + 5.7)% in AM intervention group; the EIV values were comparable between the 3 groups. After incubation with AM, the maximum EDV values of aortic rings obtained from the obese group were significantly increased from (55.6 - or + 8.3)% to (85.1 - or + 4.5)%.

CONCLUSIONAM can improve endothelial dysfunction in obese rats, and the mechanism involves improved insulin resistance and increased endothelium-derived NO productor function.

Animals ; Aorta, Thoracic ; pathology ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Endothelium, Vascular ; drug effects ; pathology ; physiopathology ; Endothelium-Dependent Relaxing Factors ; therapeutic use ; In Vitro Techniques ; Insulin Resistance ; Male ; Obesity ; physiopathology ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Vasodilator Agents ; pharmacology