Objective To detect PAH gene mutations in classical PKU patients by HRM analysis. MethodsMutation scanning of PAH gene were performed in 17 classical PKU patients by HRM analysis ( LightScanner), covering the 13 exons and exon-intron boundaries. The HRM results were further confirmed by DNA sequencing, and the sensitivity and specificity of HRM method in PKU diagnosis were also evaluated. In addition, prenatal diagnosis was performed in two fetuses at risk for classical PKU. Results In the 17 patients, two mutations were identified in 16 patients, three mutations were identified in 1 patient.In this subject, a total of 22 different pathogenic mutations : 194V( c. 280A ＞ G), IVS4nt-1 G ＞ A( c. 442-1G ＞ A), R158Q( c. 4736 ＞ A), Q160X( c. 478C ＞ T), W187X( c. 561G ＞ A), E6nt-96A ＞ G( c. 611A ＞G), G239D( c. 716G ＞ A), R241 C( c. 721C ＞ T), R243Q( c. 728G ＞ A), G247R (c. 739G ＞ C), G247V (c. 740G＞T), R261X(c. 781C ＞T), PR261Q(c. 782G ＞ A), H264R (c. 791A ＞ G), F302fsX39 (c. 904delT), E305K( c. 913G ＞ A), G312V( c. 935G ＞ T), Y356X( c. 1068C ＞ A ), V399V ( c. 1197A ＞T), R408Q(c. 1223G ＞ A), T418P(c. 1252A ＞ C) , A434D(c. 1301C ＞ A), 3 silent mutations Q232Q (c. 696G ＞ A), V245V(c. 735G ＞ A), L385L(c. 1155C ＞ G), and one single nucleotide polymorphism rs2280615 ( c. 402A ＞ C) were identified, of which 194V ( c. 280A ＞ G), Q160X ( c. 478C ＞ T), H264R (c. 791A ＞ G), G312V( c. 935G ＞ T) and E305K ( c. 913G ＞ A) were novel mutations identified in PAH gene. The prenatal diagnosis results of the two fetuses : one was diagnosed as normal, the other was diagnosed as a carrier. In this study, the sensitivity and specificity for mutation detection by HRM were 100％, and the HRM results were consistent with DNA sequencing results. Conclusions HRM analysis is a simple,accurate, rapid, high-throughput and low-cost genetic analysis approach. It could be applied to mutation scanning of classical PKU of PAH gene and rapid prenatal diagnosis in parents with known mutations.

A rapid, reliable and sensitive pressurized capillary electrochromatography-Laser induced fluorescence ( pCEC/LIF ) method with trifluoroacetic acid ( TFA ) pre-column derivation for simultaneous determination of four aflatoxin ( AFB1 , AFB2 , AFG1 , AFG2 ) was developed. This method included separation on a capillary column packed with 1. 8μm C18 particles using 0. 05% FA aqueous solution/methanol (55:45, V/V) as mobile phase at a pump flow rate of 0. 05 mL/min when the split ratio was 1:300. Under the optimum conditions including running voltage of 15 kV, excitation wavelength of 375 nm and emission wavelength of 450 nm, the baseline separation of four aflatoxins was achieved within 10 minutes. The limits of detection (LODs) were 0. 02, 0. 016, 0. 008 and 0. 01 μg/L for AFG1, AFB1, AFG2, AFB2(S/N=3), respectively. The linear detection ranges of AFG1 , AFB1 , AFG2 , AFB2 were 0. 1-10, 0. 1-10, 0. 1-3 and 0. 1-3 μg/L with correlation coefficients (R2) of 0. 9999, 1. 0000, 0. 9995 and 0. 9997, respectively. The established method was applied to analyze the peanut butter, and the recoveries of standard addition experiment were between 90 . 0% and 112 . 0% for all analytes ( RSDs=0 . 5%-1 . 9%) .

Objective To analyze the research status of private hospitals in China for policy recommendations of their development.Methods Using CNKI as the object to retrieve,and key words ofprivate hospital ,social capital to run hospitals or the titles of'private hospitals ,social capital to run hospitals to retrieve 853 articles that are valid and published since end of 2013,for a biliometric analysis.Results The number of articles published were increasing year by year;the most of the articles published in the developed southern areas;the number of articles in hospital internal management was the largest (384 articles,45.02%),followed those on external management and development situation and countermeasures (accounting for 27.43% and 22.39% respectively).Conclusion The increase of articles on private hospitals is connected with the support of national policy,the research of private hospitals is not in balance and its development environment is not favorable.

Objective To explore the perioperative experiences of congenital heart disease in infants.Methods From Jan.2000 to Aug.2006,109 patients with congenital heart disease were operated in our department,their clinical data were retrospectively collected and analyzed.The patients′ age ranged from 31 days to 3 years old (13.6 months).The body weight ranged from 2.1 to 16 kg (8.6 kg).Ninety-three patients were operated under hypothermic anaesthesia with cardiopulmonary bypass(CPB).Sixteen patients underwent deep thermal and low flow CPB.Ultrafiltration was used in 62 patients.Results There were 8 deaths and the operative mortality was 7.3%,4 cases caused by low output syndromeclos(LOS),3 cases caused by pulmonary hypertension and 1 case caused by lung intection.The morbidity was in 25 cases(22.9%),the main complications were LOS in 6 patients and respiratory complications in 18 patients,hydropericardium in 1 case,respectively.Conclusion To improve the operative and CPB technique,and to improve the skills of the postoperative managements of LOS and respiratory complications are the main points in the success of the cardiac operation in infants.

Objective To investigate the role of expressions of pituitary tumor transforming gene(PTTG) and p27 in glioma,and explore the relationship between the expressions of them and cell cycle regulation.Methods Specimens of 40 patient with gliomas were divided into gradeⅠ:8 cases,gradeⅡ:10 cases,gradeⅢ:13 cases, gradeⅣ:9 cases,PTTGmRNA and p27mRNA were detected by reverse transcription polymerase chain reaction (RT-PCR),and their expressions of PTTG,P27,CDK4 protein were detected by immunostaining assay using strep- tavidin-peroxidase(SP) method.Results The expressions of PTTGmRNA in gradeⅠ～Ⅳwere (0.907?0.065),(1.109?0.083),(1.312?0.089),(1.499?0.215) respectively,there was significant difference among the groups(P

The primary object of this fundamental research was to survey the synergistic cardiovascular effects of iptakalim, a novel ATP-sensitive potassium channel (K(ATP)) opener, and clinical first-line antihypertensive drugs, such as calcium antagonists, thiazide diuretics and β receptor blockers by a 2 x 2 factorial-design experiment. It would provide a theoretical basis for the development of new combined antihypertensive therapy program after iptakalim is applied to the clinic. Amlodipine besylate, hydrochlorothiazide and propranolol were chosen as clinical first-line antihypertensive drugs. Blood pressure, heart rate (HR) and cardiac functions were observed in anesthetized normal rats by an eight-channel physiological recorder. The results showed that iptakalim monotherapy in a low dose could produce significant antihypertensive effect. There was no interaction between iptakalim and amlodipine on the maximal changes of systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial blood pressure (MABP), the left ventricular systolic pressure (LVSP), and the left ventricular end-diastolic pressure (LVEDP) (P > 0.05). However, the effects of combination iptakalim/amlodipine on the maximal changes of SBP, DBP, MABP, LVSP and LVEDP were more obvious than those of iptakalim or amlodipine monotherapy. And there was strong positive interaction between iptakalim and amlodipine on the maximal changes of HR (P>0.05). According to the maximal changes of DBP, MABP, LVSP and LVEDP (P < 0.05) of combination iptakalim with hydrochlorothiazide, there was strong positive interaction between them. But there was no interaction between iptakalim and hydrochlorothiazide on the maximal drop of SBP and HR (P > 0.05). According to the maximal drops of DBP, MABP of combination iptakalim with propranolol, there was strong positive interaction between them (P < 0.05). But there was no interaction between iptakalim and propranolol on the maximal changes of SBP, LVSP, LVEDP and HR (P > 0.05). In conclusion, it was the first time to study the effects of amlodipine, hydrochlorothiazide or propranolol, which had different mechanisms of action from iptakalim, on cardiovascular effects of iptakalim in anesthetized normal rats. This study proved that the combination of iptakalim with hydrochlorothiazide or propranolol respectively had significant synergism on lowering blood pressure, while the combination of iptakalim/amlodipine had additive action on lowering blood pressure. Meanwhile the antihypertensive effect was explicit, stable and long-lasting. Iptakalim thus appears suitable for the clinical treatment of hypertensive people who need two or more kinds of antihypertensive agents.
Amlodipine ; pharmacology ; Animals ; Antihypertensive Agents ; pharmacology ; Blood Pressure ; drug effects ; Drug Synergism ; Heart Rate ; Hydrochlorothiazide ; pharmacology ; Hypertension ; Propranolol ; pharmacology ; Propylamines ; pharmacology ; Rats

OBJECTIVETo observe the effect of natural type ginsenoside Rg2 (Rg2) and its stereoisomers [20 (R)-Rg2 and 20 (S)-Rg2] at different concentrations on oxygen-glucose deprivation/ reperfusion (OGD/R) induced cortical neuronal injury model in vitro, and to explore the mechanism, and compare their differences of action.

METHODSCortical neurons after 7-day culture were randomly divided into 5 groups, i.e., the control group, the model group, the Rg2 group, 20 (R) -Rg2 group, and 20 (S) - Rg2 group. Cortical neurons in the Rg2 group, 20 (R)-Rg2 group, and 20(S)-Rg2 group were pretreated with 20, 40, and 80 μmol/L Rg2, 20 (R) -Rg2, and 20 (S) -Rg2 for 24 h to prepare OGD/R model. The cell survival rate, the activity of Caspase-3, the intracellular Ca2+ concentration, contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected 24 h later.

RESULTSCompared with the control group, cell survival rates and activities of SOD obviously decreased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly increased with statistical difference (P < 0.05). Compared with the model group, cell survival rates and activities of SOD obviously increased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly decreased in 20 μmol/L Rg2 group, 40 μmol/L 20 (R) -Rg2 group, and 80 μmol/L 20 (S) -Rg2 group (P < 0.05). Compared with 20(S)-Rg2 group, cell survival rates increased and contents of MDA significantly decreased in 20, 40, and 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). The activity of Caspase-3 decreased and contents of SOD increased in 80 μmol/L 20 (R)-Rg2 group, and 40, 80 μmol/L Rg2 groups (P < 0.05). Ca2+ fluorescent optical gray value decreased in 40, 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). Compared with 20 (R)-Rg2 group, Ca2+ fluorescent optical gray value decreased in 80 μmol/L Rg2 group (P < 0.05); contents of SOD increased in 40 and 80 μmol/L Rg2 groups (P < 0.05); contents of MDA decreased in 20, 40, and 80 μmol/L Rg2 groups (P < 0.05).

CONCLUSIONSRg2 and its stereoisomers could improve cell vitality of cortical neurons against OGD/R induced injury. This might be related to improving anti-apoptotic capacities and antioxidant abilities, and reducing Ca2+ inflow. Besides, the neuroprotective effect of 20 (R) -Rg2 was better than that of 20 (S) -Rg2, but inferior to that of Rg2.

Antioxidants ; metabolism ; Apoptosis ; Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; Cells, Cultured ; Ginsenosides ; pharmacology ; Glucose ; Humans ; Malondialdehyde ; metabolism ; Neurons ; drug effects ; Neuroprotective Agents ; pharmacology ; Oxygen ; Random Allocation ; Reperfusion Injury ; Stereoisomerism ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the mechanism of learning and memory dysfuction in the transgenic mouse expressing human tau 40 isoform with P301L mutation (F10).

METHODSThe human tau protein expression and phosphor-tau protein levels were detected with Western blot method. The neurofibrillary tangles were observed with Bielshowsky silver stain. The behavior changes of learning and memory were observed by open field test and passive avoidance test. Acetyleholine level, activities of acetycholinesterase and choline acetyltransferase of whole brain was detected by colorimetry method. The nitric oxide level of whole brain was detected by nitrate enzyme reduction method.

RESULTSExogenous human tau gene was expressed and an elevation of phosphor-tau protein level in 7 and 3-month transgenic mice's hippocampus andcerebrocortex was observed. The neurofibrillary tangles were observed in cerebrocortex of 7-month transgenic mice; the 7-month transgenic mice also presented an evident reduction of learning and memory ability and nitric oxide level of the whole brain, but not changes in acetylcholine level, acetycholinesterase activity, choline acetyltransferase activity and expression in whole brain.

CONCLUSIONTau transgenic mice (F10) can still inherit their parents' biologiccal characters, and develop learning and memory dysfunction awnodh san obvious decrease in nitric oxide level of whole brain in the 7-month old mice, suggesting a decrease of nitric oxide level of whole brain would be involved in the mechanism of learning and memory dysfunction in these transgenic mice.

Acetylcholine ; metabolism ; Acetylcholinesterase ; metabolism ; Animals ; Brain ; physiopathology ; Choline O-Acetyltransferase ; metabolism ; Humans ; Membrane Proteins ; genetics ; Memory Disorders ; genetics ; physiopathology ; Mice ; Mice, Transgenic ; Mutation ; Nitric Oxide ; metabolism

OBJECTIVETo investigate the susceptibility to carbon tetrachloride and benzene in offspring of expanded simple tandem repeats (ESTR) mutation mice exposed to formaldehyde (FA).

METHODSF5 and F10 offspring (200 mg/m3 x 2 hours) served as H group and ICR mice were used as control group (group C). The F5 and F10 offspring were exposed to 10 ml/kg carbon tetrachloride at the doses of 0.05%, 0.50% or 5.00% for 24 hours, respectively or 500 or 1000 mg/kg benzene for 24 hours, respectively by intraperitoneal injection. Serum alanine transaminase (ALT), aspartate transaminase (AST) and the hepatic superoxide dismutase (SOD) or malondialdehyde (MDA) were detected; also the hepatic pathological changes were observed under light microscope; the micronucleus in sternum bone marrow cells as the biomarker of benzene blood toxicity were measured.

RESULTSALT and AST activities in group C of F5 mice exposed to 0.50% and 5.00% CCl4, ALT in groups C and H of F10 mice exposed to 0.05%, 0.50%, 5.00% CCl4, AST in groups C and H of F10 mice exposed to 0.50% and 5.00% CCl4 were significantly higher than those in controls, respectively (P<0.05); as compared to the control, hepatic SOD activities in group C of F5 and F10 mice exposed to 0.50% and 5.00% CCl4, in group H of F5 mice exposed to 0.50% and 5.00% CCl4 and F10 mice exposed to 5.00% CCl4 were significantly reduced, respectively (P<0.05); however, MDA contents in group C of F10 mice exposed to 0.50% and 5.00% CCl4, in group H of F5 mice exposed to 0.05% and 0.50%, 5.00% CCl4 and F10 mice exposed to 0.50% and 5.00% CCl4 were significantly increased than those in control group, respectively (P<0.05). The susceptibility to CCl4 in ESTR mutation F5 mice exposed to FA was significantly higher than that in control F5 mice, but the susceptibility to CCl4 in ESTR mutation F10 mice exposed to FA was significantly lower than that in control F10 mice. The histopathological examination showed that the injury of hepatocytes in C and H groups significantly increased CCl4 doses, and the injury of hepatocytes in H group was higher than that in C group. The micronuclear rates in C and H group mice exposed to benzene(500 mg/kg C group, F5 and F10 mice; 1000 mg/kg C group, F5 and F10 mice; 500 mg/kg H group, F5 and F10 mice; 1000 mg/kg C group, F5 and F10 mice) were 5.88 per thousand +/- 4.55 per thousand, 8.25 per thousand +/- 2.06 per thousand, 7.50 per thousand +/- 6.99 per thousand, 10.67 per thousand +/- 1.16 per thousand, 7.88 per thousand +/- 3.09 per thousand, 9.20 per thousand +/- 1.30 per thousand, 9.63 per thousand +/- 4.34 per thousand and 13.33 per thousand +/- 2.08 per thousand, respectively, which were significantly higher than those (1.13 per thousand +/- 0.35 per thousand, 1.20 per thousand +/- 0.82 per thousand, 1.25 per thousand +/- 0.46 per thousand, 1.33 per thousand +/- 1.03 per thousand) in the solvent control group (P<0.05 or P<0.01).

CONCLUSIONFA could result in the change of susceptibility to CCl4 and benzene in offspring of ESTR mutation mice. ESTR mutation may be a biomarker of the susceptibility to chemicals, but the molecular mechanisms should be investigated in the future.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Benzene ; toxicity ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; Environmental Exposure ; Female ; Formaldehyde ; toxicity ; Genetic Predisposition to Disease ; Liver ; drug effects ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred ICR ; Mutation ; Superoxide Dismutase ; metabolism ; Tandem Repeat Sequences ; genetics