To investigate the RNA interference (RNAi) effect induced by vector-derived small interfering RNA (siRNA) targeting the three gatekeeper genes (Rad52, Ku70, Ku80) and screen the more effective target sites from candidates for further research, by using siRNA design tools online, we selected 2 candidate sequences directed to every gatekeeper gene. According to the sequences, six vector-derived siRNAs (denoted psiRNA1-6) and one mocking psiRNA7 were constructed. Among them, psiRNA1 and psiRNA2 targeted Rad52, psiRNA3 and psiRNA4 to Ku70, psiRNA5 and psiRNA6 to Ku80. The mocking psiRNA7 was used as control. After sequence identification, the seven plasmids were transfected into HepG2 cell line. siRNA-induced silencing of gatekeeper genes was determined by using RT-PCR at RNA level and Western Blot at protein level. The results showed that the six plasmids specifically targeting the coding region of gatekeeper genes were successfully designed and constructed. To some extent, the six plasmids could reduce the expression of target gene. Comparatively, the plasmid-derived siRNA psiRNA1, psiRNA4 and psiRNA5 were more effective than their counterparts. The results suggest that the gene silencing efficiency of siRNA is different, depending on their targeted region, and siRNA may provide us with practical tools for further study on the three gatekeeper genes, i.e. Rad52, Ku70, Ku80.

ObjectiveTo observe the hong-huang antioxidant on oxidative stress in patients with breast cancer during chemotherapy, including their related blood indexes, blood rheology changes, and the effects on TCM clinical symptoms and symptoms of stress.MethodsA total of 60 cases of breast cancer patients during chemotherapy from Jiangsu Province Hospital of TCM was randomly divided into treatment group and control group, 30 cases in each group. On the basis of conventional therapy, patients in treatment group were given hong-huang antioxidant (100 mL per bag) from the 1st day to the 14th day of chemotherapy, 2 bags for each day (morning and evening). Patients in control group were given foundation treatment the same as the treatment group. Patients in the two groups had their serum NO, the content of SOD, and blood rheology tested on the day before chemotherapy, and the 4th, 7th, 14th days during chemotherapy. Meanwhile, their symptom score and the integral of stress reaction and TCM symptoms were also assessed. ResultsOn the 4th day, serum NO of treatment group decreased, while SOD content increased,without statistical significance between the two groups (P>0.05). Serum NO on the 7th, 14th days was significantly lower than that in the control group, but the content of SOD was higher than that in the control group, with statistical significance between the two groups (P<0.05). Hemorheology on the 4th day significantly decreased after treatment (P<0.05), and was significantly better than that in the control group (P<0.05); Clinical symptoms and stress symptoms integral in the treatment group were significantly lower than those in the control group on the 4th, 7th, 14th days of chemotherapy, with statistical significance (P<0.05).Conclusion Hong-huang antioxidant can significantly improve the oxidative stress status, serological indexes, related blood rheology indexes, and clinical symptoms in patients with breast cancer.

Adenocarcinoma ; pathology ; Adult ; Aged ; Female ; Humans ; Lasers ; Male ; Middle Aged ; Neuroendocrine Cells ; pathology ; Polymerase Chain Reaction ; methods ; Stomach Neoplasms ; pathology

BACKGROUND:Studies have found that adipose-derived stem cells (ADSCs)/collagen complexes can promote the ADSCs differentiation and maturation into mature adipocytes and promote angiogenesis.OBJECTIVE:To explore the biological properties of the ADSCs/collagen sponge composite material and to detect its effect on angiogenesis.METHODS:(1) ADSCs were cultured on collagen sponge (experimental group) or cultured alone (control group).After 24 hours of culture,cell adhesive rate of ADSCs was determined with flow cytometry.After 2,4,6 days of culture,cell proliferation and level of vascular endothelial growth factor (VEGF) in the culture medium were detected.(2) Chick embryo chorioallantoic membranes were exposed and incubated for 7 days and then divided into four groups:0.2 mL of sterile PBS was added in the blank group,0.2 mL of 2× 108/L passage 3 ADSCs suspension was added in the ADSCs group,collagen sponge was added in the collagen sponge group,and collagen sponge with 0.2 mL of 2× 108/L passage 3 ADSCs suspension was added in the composite group.After 7 days of incubation,the microvessel count around the chorioallantoic membrane was measured.RESULTS AND CONCLUSION:(1) The cell adhesive rate of ADSCs to collagen sponge reached to (93.04±0.67)％.(2)The absorbance value (at 6 days of culture) and level of VEGF (at 4 and 6 days of culture) in the experimental group were significantly higher than those in the control group (P ＜ 0.01 or P ＜ 0.05).(3) Compared with the blank group,the number of microvessels was significantly higher in the ADSCs,collagen sponge and composite groups (P ＜ 0.05).Moreover,higher amount of microvessels were found in the composite group than the ADSCs and collagen sponge groups (P ＜ 0.05).To conclude,ADSCs can adhere well to the collagen sponge with good biocompatibility and their combined use can improve angiogenesis further by enhancing cell proliferation and VEGF secretion of ADSCs.

Objective To explore the associations between microsatellites in gonadal receptor genes and dysfunctional attitudes in adolescent with major depressive disorder(MDD).Methods Polymerase chain reaction(PCR),capillary electrophoresis and genetic scanning were performed in testing the length of microsatellites in first-onset adolescent depressive patients.Dysfunctional attitudes scale(DAS) was used in rating the dysfunctional cognition of adolescent depressive sample.These results were tested by correlative analysis and comparison analysis.Results 1.There existed significantly negative correlation between microsatellite′s length in estrogen receptor ?(ER?) gene and total score of DAS in female adolescent patients with first-onset depressive disorder.2.DAS′ total score of shorter alleles′ group was significantly higher than that of longer alleles′ group on female′ estrogen receptor ?(ER?) Gene.Conclusion The microsatellite′s length of ER? and ER? gene may have associations with dysfunctional attitudes of female adolescent with MDD.

AIMTo study the binding of sibutramine hydrochloride (SH) and bovine serum albumin (BSA) in physiological condition by spectroscopic method.

METHODSThe quenching mechanism of the fluorescence of bovine serum albumin by sibutramine hydrochloride was studied with the fluorescence and the absorption spectroscopy. The binding constants K and the number of binding sites were determined at different temperatures according to Scatchard equation and the main binding force was discussed by thermodynamic equations. The effect of the drug on bovine serum albumin conformation was also studied by using synchronous fluorescence spectroscopy.

RESULTSThe quenching mechanism of sibutramine hydrochloride to bovine serum albumin was static quenching. The binding constants K at 8 degrees C, 25 degrees C, 37 degrees C were 1.21 x 10(5), 8.31 x 10(4), 6.97 x 10(4) L x mol(-1) with one binding site, respectively. The thermodynamic parameters of the reaction were deltaH = -9.70 kJ x mol(-1), deltaS = 56.41 J x mol(-1) x K(-1).

CONCLUSIONThe binding force is electrostatic interaction. Sibutramine hydrochloride can be deposited and transported by serum protein in vivo. Sibutramine hydrochloride has nearly no effect on the serum protein conformation.

Animals ; Binding Sites ; Cattle ; Cyclobutanes ; metabolism ; Protein Binding ; Serum Albumin, Bovine ; metabolism ; Spectrometry, Fluorescence ; methods ; Spectrophotometry, Ultraviolet ; methods

OBJECTIVETo observe the characteristics and difference of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues.

METHODSUsing serial analysis of gene expression (SAGE), two SAGE libraries were generated. Forty clones from each SAGE library were sequenced, and the results were analyzed by SAGE2000 software and compared with the SAGE map at NCBI.

RESULTSA total of 655 gene tags, representing 43 genes, were extracted from the 40 sequence files of the para-tumor normal pituitary tissues and 737 gene tags, representing 53 genes, were extracted from the 40 sequence files of the pituitary adenomas. Of these tags, 13 were not reported before. The genes related to pituitary hormone secretion and energy metabolism were highly expressed in the two kinds of tissues. Some growth factors and cytokines were also expressed, including those involved in the immunological system. But there were also much difference of gene expression in the two tissues. Thirty-one and five tags were only detected in para-tumor normal pituitary tissues and pituitary adenomas, respectively.

CONCLUSIONSGenes involved in hormones secretion and energy metabolism were highly expressed in the pituitary adenomas and para-tumor normal pituitary tissues. Many growth factors and cytokines were also expressed in pituitary. There was also much difference of gene expression in the two kinds of tissues. SAGE can be used not only in understanding the quantity information of gene expression, but also in finding new genes.

Adenoma ; genetics ; metabolism ; Base Sequence ; Cloning, Molecular ; Expressed Sequence Tags ; Gene Expression ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Pituitary Gland ; metabolism ; Pituitary Neoplasms ; genetics ; metabolism

OBJECTIVETo analysis the risk factors predicting intracardial thrombus after prosthetic valve replacement.

METHODSThe clinical data of 29 cases from January 2005 to April 2009 with intracardial thrombus after prosthetic valve replacement during a 1-year follow-up was retrospectively analyzed. There were 11 male and 18 female, aged from 12 to 70 years with a mean of 48 years. The risk factors of intracardial thrombus were examined by univariate and multivariate analysis.

RESULTSUnivariate analysis found that bioprosthetic valve replacement, anticoagulation using aspirin, valve replacement at mitral position, atrial fibrillation, preoperative and postoperative internal diameter of left atrium, postoperative fibrinogen were predict factors of intracardial thrombus after prosthetic valve replacement (P < 0.05). Logistic regression analysis showed valve replacement at mitral position (OR = 9.815, P < 0.05), atrial fibrillation (OR = 5.267, P < 0.05), preoperative internal diameter of left atrium (OR = 4.529, P < 0.05) were significant risk factors of intracardial thrombus after prosthetic valve replacement.

CONCLUSIONSValve replacement at mitral position, atrial fibrillation, and preoperative internal diameter of left atrium are the correlated risk factors of intracardial thrombus after prosthetic valve replacement. Anticoagulation after prosthetic valve (especially bioprosthetic valve) replacement should be standardized to prevent intracardial thrombus formation.

Adolescent ; Adult ; Aged ; Child ; Female ; Follow-Up Studies ; Heart Diseases ; etiology ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Thrombosis ; etiology ; Young Adult

OBJECTIVETo analysis the causes of valve prosthesis-patient mismatch (PPM) after mitral valve replacement in Chinese patients.

METHODSConsecutive 100 patients for elective mitral valve replacement from January 2009 to June 2009 were enrolled and followed for this study. There were 37 males and 63 females. The mean age at operation was (52 ± 9) years (ranging 32 to 76 years). The predominant mitral valve lesion was stenosis in 60 patients, regurgitation in 14 patients and mixed in 26 patients. Among them, 63 patients were combined tricuspid valve regurgitation. Mitral valve effective orifice area was measured by Doppler echocardiography in 100 patients who received mitral valve replacement and indexed for body surface area (EOAI). PPM was defined as not clinically significant if the EOAI was above 1.2 cm(2)/m(2), as moderate if it was >0.9 and ≤ 1.2 cm(2)/m(2), and as severe if it was ≤ 0.9 cm(2)/m(2). By using the criteria, all 100 patients were classified to two groups: PPM group and no PPM group. The clinical characteristic of the patients between the two groups was compared to determine the causes of PPM and the predictors of outcomes after mitral valve replacement, such as the gender, age, valve prosthesis type, size, body surface area, and mitral valve lesion, et al.

RESULTSOf the 100 patients after MVR, 52 (52.0%) had significant PPM, 51 (51.0%) had moderate PPM, and 1 (1.0%) had severe PPM. In comparison to patients in no PPM group, patients in PPM group had a significantly larger body surface area [(1.76 ± 0.17) m(2) vs. (1.59 ± 0.13) m(2), P < 0.01] and higher prevalence of male gender (55.8% vs. 16.6%, P < 0.01). The other preoperative and operative data were similar in both groups, such as the valve prosthesis type, size, and mitral valve lesion, et al. There were no significant differences in postoperative Doppler-echocardiographic data of cardiac structure and heart function between the two groups (P > 0.05).

CONCLUSIONSThe higher incidence of PPM in mitral valve position was in male or large body surface area patients. At the time of operation, surgeons should consider the related factors, such as the patient's gender and body surface area, et al. A larger prosthesis size might be implanted to avoid PPM in mitral valve position.

Adult ; Aged ; Female ; Heart Valve Prosthesis ; adverse effects ; Heart Valve Prosthesis Implantation ; adverse effects ; methods ; Humans ; Male ; Middle Aged ; Mitral Valve ; surgery

OBJECTIVETo investigate the effects of two fluid resuscitations on the bacterial translocation and the inflammatory factors of small intestine in rats with hemorrhagic shock.

METHODSFifty SD healthy male rats were randomly divided into 5 groups (n equal to 10 per group): Group A (Sham group), Group B (Ringer's solution for 1 h), Group C (Ringer's solution for 24 h), Group D (hydroxyethyl starch for 1 h) and Group E ((hydroxyethyl starch for 24 h). A model of rats with hemorrhagic shock was established. The bacterial translocation in liver, content of tumor necrosis factor-alpha (TNF-alpha) and changes of myeloperoxidase enzyme (MPO) activities in small intestine were pathologically investigated after these two fluid resuscitations, respectively.

RESULTSThe bacterial translocation and the expression of TNF-alpha in the small intestine were detected at 1 h and 24 h after fluid resuscitation. There were significant increase in the number of translocated bacteria, TNF-alpha and MPO activities in Group C compared with Group B, significant decrease in Group E compared with Group D and in Group B compared with Group D. The number of translocated bacteria and TNF-alpha expression significantly decreased in Group E as compared with Group C.

CONCLUSIONSThe bacterial translocation and the expression of TNF-alpha in the small intestine exist 24 h after fluid resuscitation. 6% hydroxyethyl starch can improve the intestinal mucosa barrier function better than the Ringer's solution.

Animals ; Bacterial Translocation ; drug effects ; Fluid Therapy ; Hydroxyethyl Starch Derivatives ; administration & dosage ; pharmacology ; Intestine, Small ; metabolism ; Isotonic Solutions ; administration & dosage ; pharmacology ; Male ; Peroxidase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; therapy ; Tumor Necrosis Factor-alpha ; metabolism