Female ; Humans ; Infant ; Mitochondrial Proton-Translocating ATPases ; deficiency ; genetics ; Mutation

Animals ; Bufo bufo ; Caspase 3 ; genetics ; metabolism ; Female ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

Background Retinal neovascularization varies with the change of microRNA (miRNA) expression level.Quantitative real-time PCR (qRT-PCR) is a common method for the analysis of miRNA expression profiling.However,the housekeeping genes selected as references may exhibit differential expression levels under the distinctive experimental conditions.The accuracy of the levels of target gene expression often is affected if the selected housekeeping genes with significantly fluctuating expression as references.Determining a reference gene with stable expression level is the premise to consecutive studies.Objective This study was to compare the expression levels and stability of the frequently used reference genes for miRNA expression analysis in normally developing eyes and in eyes of a mouse model of oxygen-induced retinopathy (OIR),and select the optimal reference gene (s) exhibiting stable ocular expression under both hypoxia and normal development conditions.Methods P7,P12,P17,P37 and 8-week-old clean C57BL/6J mice were selected randomly.q-PCR was used to detect the dynamic changes of relative expressions of 5 kinds of reference genes in different ages of mice,including snRNAU6 (RNU6),5S ribosomal RNA (5s rRNA),snoRNA U68 (U68),snoRNA U70 (U70),snoRNA U49A (U49A).Other 40 P7 mice were randomized into the normal control group and the OIR group.The mice of the normal control group were fed in the normal oxygen environment,and those in the OIR group were raised in the (75-±2)％ oxygen environment for 10 days.Fluorescine was injected via ritrobulbar vein for retinal stretched preparation,and the histopathological examination of mouse retinas was performed to identified the models.The expressing difference of 5 kinds of reference genes in the retinas were detected to compare the differences of 5 kinds of genes expression between the two groups.GeNorm program was employed to compare the stability of the expressing genes.This study were approved and granted by Ethical Committee of Tianjin Medical University and met the requirements stipulated in the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research.Results Across the developmental ages,the expression levels of U68 in the eye exhibited the greatest variability,and then coming with U49A,U70,RNU6 and 5s rRNA.Significant differences were seen across the developmental ages for the expression levels of U68,U49A,U70,and RNU6 (U70:F =7.005,P =0.000 ; U68:F =10.189,P =0.000 ; U49A:F =21.134,P=0.000;RNU6:F=4.968,P=0.004).Expression of 5s rRNA showed the least variability across the developmental ages (F=2.099,P =0.107).In P17 mice of the OIR group,tortuous access vessels and non-perfusion area were found in the retinal section.Hematoxylin-eosin stain showed the neovascular sprout through across the inner limiting membrane.The relative expression of U70 exhibited the greatest variability and then were U49A,U68 and 5s rRNA in turn in P17 OIR mice,and significantly elevated expressions were found in U70,U49A,U68 and 5s rRNA genes between the OIR group and the normal control group (t =5.174,P =0.000;t =3.376,P =0.012;t =4.802,P =0.000 ;t=2.856,P=0.029).Expression of RNU6 showed the least variability between the two groups (t =2.104,P=0.065).As analyzed by GeNorm program,the stability for the five reference genes across developmental ages was 5s rRNA/RNU6＞ U70 ＞ U49A ＞ U68,and the optimal reference combination was 5s rRNA and RNU6.Whereas the stability for the OIR model was 5s rRNA/RNU6＞U68＞ U49A＞U70,and the optimal reference combination was 5s rRNA and RNU6.Conclusions Reference genes should be selected based on specific subjects and experimental conditions when qRT-PCR is used to analyze miRNA expression levels.In the OIR model,both developmental and hypoxic factors need to be considered.5s rRNA and RNU6 reference combination is the preferred option for the qRT-PCR analysis of ocular miRNA expression if available.

Objective To observe the effect of duration of carbon dioxide pneumoperitoneum on coagulation, fibrinolysis and endothelial activation in elderly patients undergoing laparoscopic cholecystectomy (LC). Methods The 45 elderly patients with cholelithiasis scheduled for LC, aged over 60 yeas, were placed in different groups respectively after surgery according to the duration of pneumoperitoneum. The duration of pneumoperitoneum was ≤60 minutes in group A (n=21),and more than 60 minutes in group B (n=24). Venous blood samples were taken on admission (baseline), at the end of surgery, the 1st, 2nd and 3rd day after surgery for determination of prothrombin time (PT), activated partial thromboplastin time (APTT), prothrombin fragment F1+2 (F1+2), antithrombin 3 (AT-Ⅲ activity), fibrinogen (Fib), tissue plasminogen activator (t-PA), plasminogen activator inhibitor type-1 (PAI-1), D-dimer (D-D), von Willebrand factor (vWF activity). Results Concerning the coagulation activation, at the 3rd postoperative day, the level of F1+2 was significantly higher in group B than in group A [(1.60±0.26) μg/L vs. (1.32±0.24) μg/L, P＜0.05]; AT-III was significantly higher in group B than in group A [(84.82%±20.21%) vs. (97.49%±16.87%), P＜0.05]. At the 2nd and 3rd postoperative day, the levels of Fib were significantly higher in group B than in group A [(3.87±0.62) g/L vs. (3.42±0.72) g/L, (3.98±0.77) g/L vs. (3.42±0.63) g/L, respectively, P＜0.05]. Concerning fibrinolysis, But at the 2nd and 3rd postoperative day, the level of PAI-1 was significantly higher in group B than in group A [(33.93±10.42) μg/L vs. (26.69±9.49) μg/L, (32.90±11.25) μg/L vs. (26.31±7.06) μg/L respectively, P＜0.05]. Concerning endothelial activation, at the 2nd and 3rd postoperative day, vWF was significantly higher in group B than in group A [(174.53%±44.03%) vs. (134.37%±37.74%), (176.31%±47.6%) vs. (131.21%±36.34%), respectively, P＜0.05]. Conclusions Marked activations of coagulation-fibrinolysis and endothelial activation are observed postoperatively in elderly patients undergoing laparoscopic cholecystectomy. Along with prolonged duration of pneumoperitoneum, more pronounced alterations of increased coagulation, reduced fibrinolysis and endothelial activation are observed, which could constitute an imbalanced situation of coagulation-fibrinolysis and increases the risk of venous thrombosis.

Objective To compare the changes in blood coagulation, fibrinolysis and endothelial damage in patients undergoing laparoscopic cholecystectomy with different durations of carbon dioxide pneumoperitoneum. Methods Sixty-four ASA Ⅰ orⅡpatients, aged 23-60 yr, weighing 45-82 kg, scheduled for elective laparoscopic cholecystectomy, were randomly divided into 3 groups according to the duration of pneumoperitoneum: duration of pneumoperitoneum ≤30 min group (group Ⅰ, n=21), 30 min ＜ duration of pneumoperitoneum ＜ 60 min (group Ⅱ, n=23) and duration of pneumoperitoneum≥ 60 min (group Ⅲ , n=20).The intra-abdominal pressure was maintained at 12-14 mm Hg. Venous blood samples were taken before surgery (baseline, T0 ),at the end of surgery(T1), and at 1, 2 and 3 d after surgery (T2-4) for determination of prothrombin time, activated partial thromboplastin time, concentrations of prothrombin fragment 1+2(F1+2), fibrinogen (Fib), tissue plasminogen activator and plasminogen activator inhibitor type-1 (PAI-1), and activities of antithrombin Ⅲ(AT-Ⅲ)and von Willebrand factor(vWF).Results Compared with groupⅠ , the vWF activity and PAI-1 concentration at T2 , concentrations of Fib, F1+2, PAI-1 and activity of vWF at T3 and concentrations of Fib and F1+2 at T4 were significantly increased, while the AT-IE activity at T3 was significantly decreased in group Ⅲ(P＜0.05) .Conclusion When the duration of pneumoperitoneum is short, no obvious changes in the blood coagulation, fibrinolysis and endothelial damage are observed postoperatively in patients undergoing laparoscopic cholecystectomy, and along with the prolongation of the duration of pneumoperitoneum, increased blood coagulation, reduced fibrinolysisand aggravated endothelial damage are observed postoperatively.

Objective To investigate the clinical and epidemiological characteristics of pulmonary thromboembolism (PTE) and deep venous thrombosis (DVT). Methods The clinical data of 114 200 inpatients from June 2002 to June 2008, including gender, age, smoking history, primary disease and risk factors, were reviewed. Results There were 1445(1.27%) cases with venous thromboembolism (VTE), while 1433(1. 25%) patients suffered from DVT and 153(0. 13%) patients suffered from PTE, 16(11. 11 %) patients were dead of PTE. Of all the DVT patients, there were 1348(94. 1%) cases with DVT of lower limbs with no significant difference between left or right lower limb (P>0. 05). There were 49(3.4%) cases with inferior vena cava, 23(1.6%) cases with cavitas pelvis veins and 13(0. 9%) cases with upper extremity veins. The peak ages of morbidity were between 51 to 60 years. Conclusions The incidence of VTE is increasing with ageing and there is no significant difference between males and females. The most common risk factors for thromboembolism include trauma, surgery, cardiac and pulmonary disease, age over 50 years, deep phlebitis, long-term smoking, cancer, pregnancy, childbirth, braking, history of VTE, etc. We may lower the incidence and mortality of VTE by strengthening prevention work according to the high risk factors.

Acute Disease ; Adult ; Aged ; Humans ; Male ; Middle Aged ; Pesticides ; poisoning ; Young Adult

Dental Bonding ; methods ; Dentin ; Dentin-Bonding Agents ; chemistry ; Electricity ; Humans

Researches show that exosome can take park in the development and progression of breast cancer by means of mediating the intercellular communication,which can promote cancer metastasis and drug resistance,thus influencing the treatment effect of patients with cancers.Exosome is closely related with clinical stage and prognosis of breast cancer,which has a potential value in the early diagnosis and biological therapy of breast cancer and provides a new hope for the treatment of breast cancer.

OBJECTIVEThis study is to establish the fingerprint and find out the common chromatographic peaks of Inula cappa by HPLC.

METHODThe HPLC analysis was performed on an Agilent Eclipse Plus C18 column (2.1 mm x 150 mm, 1.8 μm) with 0.1% fomic acid aqueous solution-0.1% fomic acid acetonitrile solution as mobile phase at a flow rate of 0.3 · mL(-1) · min(-1); The detective wavelength is 325 nm; The column temperature is 45 °C.

RESULTThe results indicated that 5 of 17 common peaks were identified . The similarity about 10 groups of Inulacappais is over 0.95.

CONCLUSIONThis method is able to be a scientific basis of quality assessment according to its convenient and reliable.