As a classic fluorescent detect technique, fluorescence resonance energy transfer (FRET) has been widely used in biological researches. Researchers have developed a series of fluorescence detect probes which were based on FRET. Caspase family plays an important role in apoptosis pathway, especially Caspase-8 which located, at the initial of death receptor mediated apoptosis pathway, whose its activation can trigger subsequent precaspases' activation and lead to apoptosis. So it is of great significance to detect the activation of Caspase-8 in apoptosis assay. In this study, a fluorescent probe based on FRET has been designed which can detect the activity change of Caspase-8 in cells. To identify the effectiveness and specificity of the probe, we measure the Caspase-8 activity under the Caspase-8 specifically activated apoptosis inducer RGD-TRAIL with the flow cytometry FRET detection platform. The results show that the probe can respond to the activity change of Caspase-8 in apoptotic cells, and the change can be quantified rapidly by flow cytometry. The study provides a more efficient and convenient detection method of Caspase-8 activity in living cells.

Hepatolenticular degeneration,namely Wilson's disease,an autosomal recessive disorder of copper metabolism.The author of this article gave a definition to toxin-pathogen,discussed the basic concept of collaterals disease briefly.And pointed out that hepatolenticular degeneration is a disease due to toxin-pathogen invading collaterals,which was caused by disorder of copper metabolism.This can result in toxin stagnating in collaterals and stagnation of blood stasis in collaterals in the initial stage,and collaterals injured at last.To improve QI-blood circulation and regulate the function of zang fu is important in order to accelerate elimination of toxin and prevent copper from stagnating. Specific therapy vary according to different clinical symptoms,such as promoting blood circulation for removing obstruction in collaterals and clearing toxin, clearing heat-fire and clearing toxin and dispersing stagnated liver-qi for promoting bile flow et al.

AIM: To investigate the urinary protein (UP) excretion in rats suffering from renal ischemia-reperfusion (I-R) and effect of nitric oxide on it. METHODS: SD rats were used to establish the renal I-R model. Sodium nitroprusside (SNP), N ?-nitro-L-arginine (L-NNA) and aminoguanidine (AG) were used to determine the effect of nitric oxide on UP excretion under renal I-R. Quantitative analysis of UP was made by chromatometry. UP species were separated by SDS-PAGE. RESULTS: Renal I-R caused significant increase in UP ( P

AIM: To study the up-regulation of inducible nitric oxide synthase (iNOS) in lung of pulmonary fibrosis and its relationship with fibrosis. METHODS: The changes of amount of iNOS positive stain cells and type Ⅰ?Ⅲ collagen were examined on the day 7, 14 and 30 after intratracheal administration of bleomycin A_5. The contents of NO-_2/NO-_3 (nitrite/nitrate) in out-flowing pulmonary blood (OPB), hydroxyproline in lung and the histological changes were detected after iNOS was blocked by aminoguanidine (AG). RESULTS: (1) The number of iNOS-positive stain cells increased significantly in BLMA_5 7 d, 14 d and 30 d groups compared with that in control group (P

Objective To investigate the clinical application of Glissonean pedicle transection method in hepatocellular carcinoma. Methods We analyzed the clinical data of 232 cases of hepatocellular carcinoma underwent hepatic resection in our department from April 2009 to April 2013. Eighty-nine cases underwent hepatic resection by Glissonean pedicle transection method, and 143 cases underwent Pringle maneuver. Results There was no significant differences in the operation time and blood loss between the two groups (P > 0.05). However, the postoperative hospital stay time and the postoperative complications in group A were better than those in group B(P<0.05). Levels of ALT and AST in group A increased significantly slowly than those in group B. The 1, 3-year disease-free survival rates in the two groups were 87.4%, 71.1% and 85.1%, 60.0%, respectively. The 1, 3-year survival rates were 93.3%, 73.5% and 93.4%, 65.0%, respectively, with no significant differences (P>0.05). Conclusion Glissonean pedicle transection method has many advantages including effectively preventing the intraoperative bleeding, maximally reserving the remaining of liver function, decreasing the rate of tumor spread and metastasis. Meanwhile, it is a safe and effective technique for hepatectomy without dissection of the hepatic artery, portal vein and bile duct in the hepatoduodenal ligament.

Objective To summarize the experience in application of amputation of secondary structures of splenic pedicle and self-made spleen-removing bag in laparoscopic splenectomy the using 3-hole method. Methods The clinical data of 11 patients receiving the procedure from June 2007 to April 2009 in our hospital were retrospectively analyzed. Results Advantages of the procedure were less bleeding, small wound, quick recovery and no occurrence of postoperative complications. Six cases had slight postoperative pain. All 11 patients were cured. Conclusion The technique of amputation of secondary structures of splenic pedicle is safe and feasible in laparoscopic splenectomy using the 3-hole method.

OBJECTIVETo detect potential mutation of EXT1 gene in a pedigree affected with multiple osteochondroma and explore its pathogenic mechanism.

METHODSThe coding regions and their flanking sequences of the EXT1/EXT2 genes were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified by excluding possible single nucleotide polymorphisms and bioinformatics analysis. Transcripts of the EXT1 gene in the proband were analyzed by TA clone-sequencing, with its abundance compared with that of healthy controls.

RESULTSDNA sequencing has identified in the proband a novel heterozygous point mutation (c.1164+1G to A) at the 5'splice sites of intron 3 of the EXT1 gene. The same mutation was not found in the healthy controls. Bioinformatics analysis indicated that the mutation is highly conserved and can lead to skipping of exon 3 or aberrant splicing. TA clone-sequencing indicated that the numbers of transcripts with skipping of exon 3 has significantly increased in the proband (< 0.05) compared with the controls.

CONCLUSIONThe c.1164+1G to A mutation has resulted in skipping of exon 3 in a proportion of EXT1 gene transcripts. As the result, the number of transcripts with tumor suppressing function is relatively reduced and has ultimately led to the tumors.

Adult ; Base Sequence ; Child ; Exostoses, Multiple Hereditary ; genetics ; Female ; Humans ; Male ; Molecular Sequence Data ; N-Acetylglucosaminyltransferases ; genetics ; Point Mutation ; RNA Splice Sites ; RNA Splicing

The occurrence of the clinical manifestations of myocardial ischemia shows clear circadian rhythmicity ,and they are unevenly distributed during the 24 h with higher morbidity during the initial hours of the daily activity span and in the late afternoon or early evening .Such temporal patterns result from circadian rhythms in pathophysiological mechanisms plus cyclic environmental stressors that trigger these clinical events .β‐receptor antagonist medications ,oral nitrate ,and calcium channel blocker have been shown to be influenced by the circadian time of their administration .Here we briefly review the char‐acteristics of circadian rhythmicity in MI ,the pathophysiological mechanisms as well as the current chronotherapy ,and then discuss the future treatment strategies .

Objective:To investigate the biomechanical properties of a new pediatric femoral neck system in the fixation of pediatric femoral neck fractures with a free fracture fragment.Methods:Ten Sawbones model bones were randomly divided into 2 even groups ( n=5), all of which were made into Delbet Type Ⅱ femoral neck fractures with a Pauwels angle of 70°. A free bone block was removed from the bottom at the proximal end of the fracture to simulate a femoral neck fracture with a free fragment. Group A were fixed with traditional inverted triangle cannulated screws, and group B with a new pediatric femoral neck system. After the 2 groups of specimens were placed on a biomechanical testing machine, each specimen was subjected to a static axial compression test, an anti-torsion test and a cyclic load test in turn. The biomechanical results were compared between groups A and B in aspects of axial compression stiffness, torsional stiffness and maximum displacement difference. Results:The axial compression stiffness [(321.718±5.770) N/mm] and torsional rigidity [(1.448±0.079) N·m/°] in group B were significantly higher than those in group A [(266.722±4.788) N/mm and (1.282±0.023) N·m/°] ( P<0.05). The maximum displacement difference in the cyclic load test in group B [(0.063±0.038) mm] was also significantly smaller than that in group A [(0.117±0.056) mm] ( P<0.05). Conclusion:In fixation of pediatric femoral neck fractures with a free fracture fragment, the new pediatric femoral neck system can lead to better biomechanical stability than the traditional inverted triangle cannulated screws.

Objective To screen potential metabolites and significantly altered metabolic pathways of liver lesions by central carbon pathway metabolites. Methods 32 healthy volunteers (HC), 23 patients with biliary cysts (CYST), 19 patients with biliary stones (Stone), 45 patients with hepatocellular carcinoma (HCC), and 50 patients with hilarcholangiocarcinoma (HCCA) were recruited. Their serum samples were collected for UPLC-QQQ-MS analysis and further MPP statistical analysis. Pattern recognition was further used to discovery the differences in metabolome between groups, and to explore the significantly altered metabolic pathway and possible pathogenic mechanism of liver diseases. Results A total of 15, 7, 7, and 3 metabolites and a total of 8, 4, 4, and 1 metabolic pathway that were significantly different in serum between CYST, Stone, HCC, HCCA and healthy controls were identified and enriched through serum metabolomics analysis, respectively. Conclusion According to the above identified differential metabolites and enriched metabolic pathway results, it is shown that liver lesions mainly involved in the energy metabolism and amino acid metabolism & transport, in addition, inositol phosphate metabolism were significantly changed both in CYST, Stone, HCC and HCCA.