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.

Objective:To develop a catalytic system for the asymmetric Morita-Baylis-Hillman ( MBH) reaction of conjugated ni-troalkene with activated aldehyde, and screen out the chiral catalysts with high activity and enantioselectivity. Methods: Totally 21 chiral organocatalysts were applied in the asymmetric MBH reaction ofβ-nitrostyrene with ethyl glyoxylate, and the ee value was deter-mined by chiral HPLC. The effects of temperature, solvent and substrate ratio on the catalytic reaction were investigated. Results: In the presence of cinchona alkaloid catalyst (DHQ)2AQN, β-nitrostyrene reacted with ethyl glyoxylate in toluene at 0℃ affording the MBH adduct in 60% yield with good enantioselectivity (up to 56.9% ee). Conclusion: The bis-cinchona alkaloids with aromatic bridging group are the efficient catalysts for the asymmetric MBH reaction ofβ-nitrostyrene with ethyl glyoxylate, and moderate isolated yield and enantioselectivity are obtained.

Objective To determine whether the poking reduction and bone grafting technique with guide through bony tunnel can correct a Hill-Sachs lesion. Methods A total of 30 cadaveric humeri were equally divided into three groups, 10 cadaveric humeri per group. Hill-Sachs lesions were replicated with a osseous defect involving 10% (group A ) , 20% (group B ) and 30% (group C ) of the articular surface. All the bone defects in each group were measured and the poking reduction and bone grafting technique with guide through a bony tunnel was performed in group B and group C. The preoperative and postoperative transverse arc length, longitudinal are length, depth and volume of the osseous defects in group B and group C were compared by using paired t test. Results Before reduction, the transverse arc length of the bone defects was ( 10.9 ± 1.4 )mm in group B and ( 16.3 ± 2.3 ) mm in group C ; longitudinal arc length was ( 22.4 ± 2.4 ) mm in group B and ( 28.0 ± 2.2 ) mm in group C ;depth was (6.9±0.9) mm in group B and (11. 1 ±0.9) mm in group C; volume was (708.7±93.9) mm3 in group B and (1338.3 ± 185.6) mm3 in group C. After reduction, the transverse arc length of the bone defects was (5.1 ± 2.4 ) mm in group B and ( 7.6 ± 3.6 ) mm in group C ; longitudinal arc lengthwas (10.5 ±4.9) mm in group B and (12.3 ±5.3) mm in group C; depth was (0.3±0.1 ) mm in group B and (0.4 ±0.1 ) mm in group C; volume was (48.9 ± 16.1 )mm3 in group B and (70.3 ± 37.9) mm3 in group C. The comparison of all the parameters showed statistical difference (P <0. 01 ). Conclusion The poking reduction and bone grafting technique with guide through a bony tunnel can effectively correct the Hill-Sachs lesions with humeral head osseous defects involving 20% -30% of the articular surface.

OBJECTIVETo analyze the complications of percutaneous kyphoplasty except bone leakge for the treatment of osteoporotic thoracolumbar vertebral compression fractures.

METHODSFrom October 2008 to October 2012,178 patients with 224 osteoporotic vertebral compression fractures were treated with percutaneous kyphoplasty under local anethsia. There were 72 males and 106 females,ranging in age from 58 to 92 years old,with an average of 75.3 years,including 93 thoracic vertebrae and 131 lumbar vertebrae. The complications except bone cement leakage were analyzed during operation and after operation.

RESULTSAll operations were successful and all patients were followed up from 12 to 60 months with an average of 26.2 months. No death was found. Bone cement leakage occurred in 27 cases, about 15.1% in 178 cases; and complications except bone cement leakage occurred in 15 cases. There was 1 case with cardiac arrest,was completely recovery by cardiopulmonary resuscitation (CPR) immediately; and 1 case with temporary absence of breathing,was recovery after treatment. There were 3 cases with fall of blood pressure and slower of heart rate; 1 case with intestinal obstruction; 2 cases with local hematoma and 1 case with intercostal neuralgia. Vertebral body fractures of 2 cases were split by bone cement and the fractures of adjacent body occurred in 4 cases.

CONCLUSIONIt's uncommon complication except bone cement leakge in treatment of osteoporotic thoracolumbar vertebral compression fractures with percutaneous kyphoplasty. The complication of cardiopulmonary system is a high risk in surgery; and cytotoxicity of bone cement,nervous reflex,fat embolism and alteration of intravertebral pressure may be main reasons.

Aged ; Aged, 80 and over ; Female ; Fractures, Compression ; surgery ; Humans ; Kyphoplasty ; adverse effects ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Osteoporotic Fractures ; surgery ; Postoperative Complications ; etiology ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery

OBJECTIVETo study the phenotypes and genotypes of a protein C (PC) deficiency pedigree.

METHODSImmunoassay (ELISA) was used for PC antigen and activated PC (APC) detection, PCR for amplification of the fragment of protein C gene exon II to exon IX, single-strand conformation polymorphism (SSCP) for difference of denatured cDNA and DNA sequencing for gene mutation.

RESULTSFour members in the pedigree were found to be PC antigen levels between 34.3% - 67.8% and PC activity between 22% - 49% which are lower in comparison with normal references (80% - 120% and 70% - 130%, respectively). A G-to-A mutation in exon VII of the protein C gene at 6 219 position was identified in 9 members. This mutation resulted in the substitution of Arg for Gln at 169 amino acid.

CONCLUSIONThe proband is of heterozygosity. The G6219 A mutation in exon VII of the protein C gene leads to the substitution of Arg 169 Gln. This mutation is reported for the first time in China.

Adult ; DNA Mutational Analysis ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; Protein C ; genetics ; metabolism ; Protein C Deficiency ; congenital ; genetics

Objective To explore the effect of Ginsenoside Rgl on glutathion (GSH) level and glutathion peroxidase (GPx) activity after oxygen-glucose deprivation/reperfusion in cultured hippocampal cells. Methods The model of oxygen-glucose deprivation and reperfusion were established with the hippocampal neurons of rats. They were randomly divided into control group, model group and Ginsenoside Rgl treatment groups (5 μmol/L, 20 μmol/L, 60 μmol/L). The GSH level and GPx activity were measured 6 h after reperfusion. The apoptosis and the metabolic rate of methyl thiazolyl tetrazolium (MTT) were detected 24 h after reperfusion. Results Compared with model group, the GSH level, GPx activity, and metabolic rate of MTT improved (P<0.001), and the apoptosis decreased in the Ginsenoside Rgl groups (P<0.001) except with the dosage of 5 μmol/L (P>0.05). Conclusion Ginsenoside Rgl can protect the brain from ischemia by increasing the GSH level and GPx activity.

Aim To explore the antidepressant effect and the mechanism of self-made prescription of strengthening spleen and replenishing Qi ( SMP-SS-RQ) . Methods The depression model of chronic un-predictable mild stress ( CUMS ) was established in mice. The animals were randomly divided into control group, model group, SMP-SSRQ low, middle and high dose groups (8, 16, 32 mg·kg-1), with 12 mice for each group. The control group and the model group were given the same volume of saline, and other groups were given corresponding dose of SMP-SSRQ. Animals of each group were administered by gavage twice a day for two weeks. Behavioral indexes of mice were deter-mined by open field experiment, sugar consumption test, forced swimming test and tail suspension test. Quantitative real-time PCR (qPCR) and Western blot were respectively performed to detect mRNA and pro-tein expression levels of Igsf11 , Pdia2 and Sec14 l2 inprefrontal cortex and hippocampus. Results Com-pared with model group, depressed mice' s horizontal mobile score, upright number and the sugar water pref-erence index increased and FST as well as TST de-creased in all SMP-SSRQ groups(P<0.05). The de-pression symptoms in mice were obviously improved by SMP-SSRQ therapy. Low and high doses of SMP-SSRQ significantly reduced mRNA and protein expression lev-el of Igsf11 , Pdia2 and Sec14 l2 in the prefrontal cortex and hippocampus of the depressed mice, presenting significant statistical difference compared with model group ( P<0.05 ) . Conclusions SMP-SSRQ can ef-fectively improve mouse depressive behavior, and its mechanism may be related to the down-regulation of Igsf11 , Pdia2 and Sec14 l2 in mouse prefrontal cortex and hippocampus.

Objective To investigate the microbleeding incidence of healthy eldery population and patients with stroke.Methods 30 cases of healthy eldery population,32 cases of cerebral hemorrhage,46 cases of patients with ischemic cerebral vascular diseases were performed of MRI and GRE-T_2 ~* WI examination.Results The microbleeding incidences was 37.5% in cerebral hemorrhage group,28.1% in multiple cerebral infarction group,25.0% in Binswanger's disease group.The most frequently seen microbleeding foci located in ganglia areas,then in thalamus areas,subcortical areas and brain stem,last in cerebellar.Conclusion GRE-T_2 ~* WI,helpful for finding microbleeding and indicating lesion degree of microblooding vessels,plays an important role in the diagnosis of stroke and decision making of treatment.

Cantharidin is a terpenoid compound secreted by Mylabrisphalerata Pallas.In a variety of cancer types,cantharidin has shown the effects of inhibiting cancer cell growth,proliferation and migration.The anticancer mechanism of cantharidin involves inducing cell cycle arrest and apoptosis,inhibiting autophagy,enhancing DNA damage,inhibiting DNA repair,and regulating various cell signaling pathways.This article reviews the mechanism and research progress of cantharidin and its derivatives in common cancer types,in order to identify novel targets of cancer treatment using cantharidin,and provide new direction for clinical research to improve anticancer therapy.

Objective To investigate the palliative care cognition of medical workers and provide evidence for medical worker' s vocational education and treatment of patients with end-stage.Methods 280medical workers from seven different hospitals in five provinces were surveyed with self-designed questionnaire of their palliative care cognition from January to March 2011.Results 280 questionnaires were handed out and 269 were recovered,with the recovery rate of 96.1％.The palliative care cognition of medical workers was affected by work type,age,working years,and working departments.Doctors and medical workers with age between 36 and 45 years old and working years more than 10 years had higher palliative care cognition,and the scores were respectively (25.81 ± 5.51),(26.88 ± 4.40) and (26.23 ± 4.92).Nurses,medical workers with age under 25 years old,working years less than 5 years and in surgical department had lower palliative care cognition,and the scores were respectively (22.25 ± 6.31),(23.68 ± 6.56),(23.57 ± 5.78),(22.55 ± 6.49).The differences were statistically significant (F =6.989,3.961,5.877,2.677,respectively; P ＜0.01).Conclusions Education of palliative care for medical workers,especially for nurses,is needed to provide high quality medical service for patients.