Animals ; Biomarkers ; Hexanes ; poisoning ; toxicity ; Humans ; Neurotoxicity Syndromes ; etiology

OBJECTIVETo investigate the possible mechanism of compound Chinese sour taste herbs (CS) in preventing and ameliorating diabetic macroangiopathy by analyzing the effects of CS on the deposition of advanced glycation end products (AGEs) and gene expression of receptor for advanced glycation end products (RAGE) in the aorta tissue of rats with type 2 diabetes mellitus (T2DM).

METHODSRat model of T2DM was established by peritoneal injection of streptozotocin (STZ) and high caloric diet feeding. Experimental SD rats were divided into the normal group, the model group, the aminoguanidine (AG) group, and the CS group. At the end of the 8th and 12th week, the fasting blood glucose (FBG) was measured by glucose oxidase method; content of AGEs and collagen in aorta detected by fluorescent method and gene expression of RAGE in aorta determined by Real-time PCR method.

RESULTSFBG, AGEs and collagen contents and RAGE expression in aorta of model rats were all higher than those in the normal control group (P <0.05), while all these indices were lower in the CS group than in the model group (P<0.05).

CONCLUSIONCS could realize the goal for preventing and ameliorating diabetic macroangiopathy by way of suppressing the production of AGEs and down-regulating the gene expression of RAGE in aorta of T2DM rats.

Animals ; Aorta ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Glycation End Products, Advanced ; metabolism ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; genetics ; metabolism

OBJECTIVETo analyze the relationship of K-ras mutation with the development of liver metastasis in colorectal cancer patients and the survival outcomes.

METHODSFrom 2003 to 2008, 300 patients who underwent colorectal cancer surgery in the Department of General Surgery of Zhongshan Hospital, Fudan University were assigned to different groups, according to the diagnosis and follow-up results. The mutation of exon 2 of K-ras was detected in primary paraffin-embedded lesions by PCR and Pyrosequencing. The association of gene mutation with the development of liver metastasis and its prognosis was studied.

RESULTSAmong 300 cases, the mutations of exon 2 were present in 120 cases(40%). The G13D mutation was more common in metachronous metastasis group than that in synchronous group(17.0% vs. 8.0%, P=0.041). Multivariable regression analysis showed that G13D mutation was an independent risk factor(HR=1.108, 95%CI:1.032-5.062, P=0.048) for metachronous metastasis. Patients with mutated K-ras had a poorer overall survival compared to those without mutated K-ras for patients without liver metastasis(median overall, 65 vs. 72 months, P=0.039), and for patients who received metastasis resection(median disease-free survival 18 vs. 24 months, P=0.048). Multivariable analysis showed that K-ras mutation was an independent risk factors of overall survival(HR=1.561, 95%CI:1.022-6.422, P=0.045) in patients without liver metastasis.

CONCLUSIONDetection of K-ras mutation may predict the development of liver metastasis and prognosis.

Aged ; Colorectal Neoplasms ; genetics ; pathology ; Female ; Genes, ras ; genetics ; Humans ; Liver Neoplasms ; genetics ; secondary ; Male ; Mutation ; Prognosis

Acute Disease ; Adolescent ; Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Pesticides ; poisoning ; Poisoning ; epidemiology

2600 Anthers from T0 modified cry1 Ac-transgenic rice lines of Minghui 81, an elite restoring line of commercial CMS indica hybrid rice, were cultured on SK3 media. 83 green plantlets were recovered, 43 double haploid (DH) and 40 haploid among them. Results of PCR analyzes indicated that 55 plants of 83 were harbored the cry1Ac gene, and the ratio of cry1Ac-positive against cry1Ac-negative was 2:1 (55/28). 36 putative transgenic DH plants were further confirmed by Southern blot. ELISA detection showed that Cry1Ac level in different transgenic rice plants of the same cry1Ac-DH clone was almost equal and the highest one amount to 0.25% of the total soluble protein. Pest insect-resistant bioassay at field trials demonstrated that some of the homozygous cry1Ac-transgenic rice plants not only showed high-level resistance against striped stem borer (Chilo suppressalis) but also retained elite agronomy characters. These results demonstrated that rice anther culture has a great value in rice molecular breeding.
Animals ; Bacillus thuringiensis ; genetics ; Bacterial Proteins ; genetics ; metabolism ; Bacterial Toxins ; Blotting, Southern ; Culture Techniques ; DNA, Plant ; genetics ; Endotoxins ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Hemolysin Proteins ; Immunity, Innate ; genetics ; Moths ; growth & development ; Oryza ; genetics ; growth & development ; parasitology ; Plant Structures ; genetics ; growth & development ; parasitology ; Plants, Genetically Modified

Prawn white spot syndrome is caused by the pathogen prawn white spot syndrome virus (WSSV). VP19 is a vesicle membrane protein of WSSV. HyNPV (Hybrid of AcNPV and BmNPV) constructed by the recombination of BmNPV and AcNPV is a new hybrid virus having both of their advantages. The recombinant transfer vector pBlueBicHisC-vp19 and recombinant baculovirus HyNPV-VP19 were constructed on the basis of the successful cloning of VP19. Newly-molted silkworms Bombyx mori of fifth instar were inoculated by the recombinant virus. SDS-PAGE and Western blotting analysis showed a specific band, about 21kD, which was consistent with the expectation suggesting that the WSSV-VP19 gene was successfully expressed in silkworm bodies.
Animals ; Baculoviridae ; genetics ; metabolism ; Bombyx ; genetics ; metabolism ; virology ; Genetic Vectors ; Penaeidae ; virology ; Recombinant Fusion Proteins ; genetics ; Viral Envelope Proteins ; genetics ; metabolism ; Virus Replication ; White spot syndrome virus 1 ; genetics

OBJECTIVETo study the sympathetic skin response (SSR) to the effects of N-hexane on autonomic nerves function in patients with chronic N-hexane poisoning.

METHODSThe subjects in present study included 30 controls and 37 cases with chronic N-hexane poisoning. Also 37 patients were divided into 3 subgroups (mild, moderate and severe poisoning) according to diagnostic criteria of occupational diseases. All subjects were examined by SSR test and nerve conduction velocity (NCV) test. All patients were reexamined by SSR and NCV every 1 ∼ 2 months. The differences in SSR parameters (latency, amplitude) among groups were observed. In the severe poisoning subgroup, the changes of SSR and NCV parameters (conduction velocity, amplitude) in different poisoning stages were observed.

RESULTSThere were significant differences in SSR latency of upper extremity among groups and the significant differences in SSR amplitude of upper and lower extremity among groups (P < 0.05). No significant differences in SSR parameters were found between the adjacent groups (P > 0.05). There were significant differences in SSR latency of upper extremity during different periods and the significant differences in SSR amplitude of upper and lower extremity during different periods among all groups (P < 0.05). The change of SSR parameters consistent with that in NCV. The longest SSR latency of upper extremity and the smallest SSR amplitudes of upper and lower extremity appears 1 - 2 months earlier than that of the smallest action potential amplitude.

CONCLUSIONThe damage of autonomic nerves induced by N-hexane increased with poisoning progresses. The damage of autonomic nerves corresponded with the damage of myelin sheath of large myelinated nerves, but which appeared 1 - 2 months earlier than the damage of axon of large myelinated nerves. SSR test may serve as a method to detect the damage of autonomic nerves function in patients with chronic N-hexane poisoning.

Adolescent ; Adult ; Autonomic Pathways ; physiopathology ; Case-Control Studies ; Female ; Galvanic Skin Response ; Hexanes ; poisoning ; Humans ; Male ; Neural Conduction ; Occupational Diseases ; physiopathology ; Skin ; innervation ; physiopathology ; Sympathetic Nervous System ; physiopathology ; Young Adult

OBJECTIVETo study the effect of 5-fluorouracil-FU in combination with astragalus membranaceus(AM) on amino acid metabolism in mice model of gastric carcinoma induced by 3-methylcholanthrene(MC).

METHODSMice gastric carcinoma models were established by 3-methylcholanthrene induction and randomly divided into different groups, and received 5-FU treatment (group A) 5-FU plus AM (group B), 5-FU plus a high dose of AM(group C), no treatment (group D). Normal mice were used as control (group N). Free amino acid in the tumor specimens were examined.

RESULTSThe levels of free Valine, Methionine, Leucine, Arginine and cystine in the tumor specimens in group D were significantly higher than that in group N(P< 0.05). The levels of free serine in group A, B, C, D were significantly higher than that in group N. The levels of free glutamic acid in group A, B were significantly higher than that in group N(P< 0.05). The levels of free proline in group C, D were significantly higher than that in group P, N(P< 0.05).

CONCLUSIONSThe increasing levels of free serine and proline in tumor specimens in gastric cancer mice model reveals metabolic disturbance of amino acid. 5-FU plus astragalus membranaceus can decrease the level of free glutamic acid in the mice models, and inhibit tumor growth.

Amino Acids ; metabolism ; Animals ; Astragalus membranaceus ; Fluorouracil ; therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Phytotherapy ; Stomach Neoplasms ; drug therapy ; metabolism

Objective To discuss the characteristic of brachial plexus root avulsion injury of high resolution MR imaging and the value in diagnosing of brachial plexus root avulsion injury early.Methods Fourty-five cases of brachial plexus root avulsion injury patients had being used for investigation to find the characteristic and diagnostic value of MR image of brachial plexus root avulsion injury,which all have pre-operative MR imaging and were diagnosed brachial plexus root avulsion injury by intra-operative exploration and electrophysiology form February 2006 to February 2011.Results Post-traumatic spinalmeningolceles were seen in 42 cases,the frequency was 93.3％; Displacement of spinal cord was seen in 25 cases,the frequency was 55.6％; Absence of anterior and posterior root of spinal nerve was seen in 8 eases,the frequency was 17.8％;Black line sign was seen in 18 cases,the frequency was 40.0％.The sensitivity,specificity,and accuracy of MRI in diagnosing brachial plexus root injury were 95.7％,77.8％ and 94.6％ respectively.Conclusion Posttraumatic spinalmeningolceles are most often seen in MR of brachial plexus root avulsion injury,this sign can play an important role in diagnosing and treatment of brachial plexus root avulsion injury.

In our recent study by exploring an intein-based dual-vector to deliver a B-domain-deleted FVIII (BDD-FVIII) gene, it showed that covalently ligated intact BDD-FVIII molecules with a specific coagulant activity could be produced from expressed heavy and light chains by protein trans-splicing. Here, we assessed the hypothesis that the efficiency of trans-splicing may be increased by adding to the intein sequences a pair of leucine zippers that are known to bring about specific and strong protein binding. The intein-fused heavy and light chain genes were co-transferred into cultured COS-7 cells using a dual-vector system. After transient expression, the intracellular BDD-FVIII splicing was observed and the spliced BDD-FVIII and bioactivity secreted to culture media were quantitatively analyzed. An enhanced splicing of BDD-FVIII with decreased protein precursors from gene co-transfected cells was observed by Western blotting. The amount of spliced BDD-FVIII and bioactivity secreted to the culture media were 106 +/- 12 ng x mL(-1) and 0.89 +/- 0.11 U x mL(-1) analyzed by ELISA and Coatest method respectively, which was greater than leucine zipper free intein-fused heavy and light chain genes co-transfected cells (72 +/- 10 ng x mL(-1) and 0.62 +/- 0.07 U x mL(-1)). The activity of cellular mechanism-independent protein splicing was also improved, as showed by the increasing of spliced BDD-FVIII and bioactivity in culture media from combined cells separately transfected with heavy and light chain genes which was 36 +/- 11 ng x mL(-1) and 0.28 +/- 0.09 U x mL(-1). It demonstrated that the leucine zippers could be used to increase the efficiency of protein trans-splicing to improve the efficacy of a dual-vector mediated BDD-FVIII gene delivery by strengthening the interaction between the two intein-pieces fused to heavy and light chains. It provided evidence for further study in animal model using a dual-adeno-associated virus vector to deliver FVIII gene in vivo.
Animals ; COS Cells ; Cercopithecus aethiops ; Factor VIII ; chemistry ; genetics ; metabolism ; Genetic Vectors ; Inteins ; Leucine Zippers ; Peptide Fragments ; chemistry ; genetics ; metabolism ; Protein Splicing ; Trans-Splicing ; Transfection