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The systemicity of chlorantraniliprole was investigated and evidence was collected for formulating strategies in controlling vegetable or rice pests. Systemicity of chlorantraniliprole was investigated through chlorantraniliprole application on velvetleaf (Abutilon theophrasti) leaves as well as through hydroponic experiments. The roots of velvetleaf were incubated in chlorantraniliprole solution with a concentration of 50 microg/mL; the concentrations of chlorantraniliprole in xylem above the solution parts were 3.14, 5.67, and 6.89 microg/g at 24, 48, and 72 h after treatment. When the roots were incubated in chlorantraniliprole solution with a concentration of 200 microg/mL, the concentrations of chlorantraniliprole in xylem above the solution reached 6.48, 8.76, and 10.55 microg/g at 24, 48, and 72 h after treatment. Chlorantraniliprole was not detected in the phloem above the solution after these two treatments. When chlorantraniliprole solution with a concentration of 100 microg/mL was applied on mature leaves, chlorantraniliprole was found in xylem above the mature leaves at concentrations of 0.55, 0.74, and 0.92 microg/g at 24, 48, and 72 h after the treatment. No chlorantraniliprole was detected in the leaves below the mature leaves or the phloem above them. When chlorantraniliprole solution with a concentration of 100 microg/mL was applied on apical leaves, no chlorantraniliprole was detected in the xylem or phloem below them. These results indicated that chlorantraniliprole can be transported through xylem only upward, but chlorantraniliprole has no phloem mobility in velvetleaf.

To study the effect of VEGF overexpression on development of cortical newborn neurons in the brains after stroke, we injected human VEGF(165)-expressive plasmids (phVEGF) into the lateral ventricle of rat brains with a transient middle cerebral artery occlusion (MCAO). An injection of phVEGF significantly promoted angiogenesis (BrdU(+)-von Willebrand's factor(+)) and reduced infarct volume in the rat brain after MCAO. Single labeling of 5'-bromodeoxyuridine (BrdU) and double staining of BrdU with lineage-specific neuronal markers were used to indicate the proliferated cells and maturation of newborn neurons in the brain section of rats at 2, 4, and 8 weeks after MCAO. The results showed that BrdU positive (BrdU(+)) cells existed in ipsilateral frontal cortex within 8 weeks after MCAO and reached the maximum at 2 weeks of reperfusion. The phVEGF treatment significantly increased BrdU(+) cells compared with the control plasmid (pEGFP) injection. Cortical neurogenesis was indicated by the presence of newborn immature (BrdU(+)-Tuj1(+)), newborn mature (BrdU(+)-MAP-2(+)), and newborn GABAergic (BrdU(+)-GAD67(+)) neurons. All these neurons declined within 8 weeks after MCAO in the controls. Injection of phVEGF significantly increased BrdU(+)-Tuj1(+) neurons at 2 weeks, and BrdU(+)-MAP-2(+) neurons and BrdU(+)-GAD67(+) neurons at 4 and 8 weeks, respectively after MCAO. Moreover, phVEGF treatment significantly increased neurite length and branch numbers of BrdU(+)-MAP-2(+) newborn neurons compared with pEGFP treatment. These results demonstrate that VEGF enhances maturation of stroke-induced cortical neurogenesis and dendritic formation of newborn neurons in adult mammalian brains.

Objective: To study the anatomy of peroneal tendofascial flap combined with adipofascial flap for the repair of heel tissue defects., Methods: The lower extremities of five cadavers (10 sides) were perfused with red latex, the blood supply of peroneal tendofascial flap and vicinity adipofascial flap were observed. The diameter, course, branches and location of the blood vessels were measured. Eight fresh cadavers (16 sides) were perfused with lead oxide-gelatine mixture. The covering fascia tissues of the lower extremities was obtained and photographed by X-ray. The vascular anastomosis and association of nutrient vessel of peroneal tendofascial flap and vicinity adipofascial flap were observed. Two adult lower extremities specimens (4 sides) were used to construct vessel diagrams for observation of the course, distribution and anastomosis of the vessels. Eight cases were treated successfully with theses flaps., Results: The blood supply of the combined fascial flap is multi-originated. For the area within 4 cm below and above the lateral malleolus cusp, the blood supply includes 2-5 branches from heel lateral artery with an average diameter of (0.5 +/- 0.2) mm, 1-2 branches from posterior lateral malleolus artery with an average diameter of (0.6 +/- 0.2) mm and 2-3 branches from the descending part of perforating branches of peroneal artery with an average diameter of (0.5 +/- 0.2) mm. The blood supply of area 4 cm above lateral malleolus cusp is 1-3 branches from intermuscular septum perforating branches of peroneal artery with an average diameter of (1.0 +/- 0.2) mm. These above branches are anastomosed each other and also send off many smaller branches to form vascular net around tendon. The fascial flaps and free skin grafts in eight patients were completely survived. All patients were followed up for 3-24 months, the donor and recipient sites were healed very well. The functional and cosmetic results were satisfactory., Conclusions: Peroneal tendofascial flap combined with adipofascial flap, with proximal pedicle or reverse distal pedicle, can be used to repair the defect at the lower leg and refractory small- and medium-sized defects at the heel.

A quaternary ammonium chitosan, 2-hydroxypropyltrimethylammonium chloride chitosan (HACC), has been developed for the dynamic coating material in CE for the first time. It presented many advantages such as favorable water solubility, satisfactory coating efficiency, and EOF toward the anode at pH >7.0. Using the modified fused-silica capillary, sulfonamides (SAs), an important group of veterinary drugs, were separated and detected by CE combined with field-amplified sample injection (FASI). The LODs of sulfonamides with UV detection were less than 0.5 ng/mL. The proposed method has been applied to the determination of veterinary sulfonamide residues in samples such as chicken, beef, and honey with fast separation (15 sulfonamides within 20 min), low LODs (0.1-0.5 ng/mL), and good reliability compared to the criteria of China (GB/T 18932.17-2003).

Totally 2803 SSRs distributed in 2443 ESTs were mined out and accounted for 13.58% of 17987 non-redundant ESTs from oilseeed rape, with the average distance of distribution about 4.26 kb. Dinucleotide and trinucleotide repeats are the dominant type, with similar frequency and accounting for 89.05% together in all SSRs. AG/CT and AAG/CTT are the most frequent motifs, accounting for 84.31% and 37.71% in dinucleotide and trinucleotide repeats respectively. Further, 23 primer pairs for EST-SSRs were designed and the suitable annealing temperature for each primer pair was determined by gradient PCR. The amplification and polymorphism displayed by these primers in 10 varieties of oilseed rape were detected by using silver staining of nondenaturing polyacrylamide gel. 21 primer pairs showed the amplification, accounting for 91.30% of total primers, and 12 primer sets showed polymorphisms, accounting for 57.14% of primers available. These results indicate that it is an effective and feasible approach to develop SSR markers based on ESTs in oilseed rape.

Forty two tea varieties were analyzed by using 16 SSR primer sets derived from tea ESTs in this study, and 13 of the primer sets produced clear bands and 10 of them showed polymorphism, accounting for 76.9%. The PIC (polymorphism information content) for each polymorphic primer set varied from 0.522 to 0.866, with a average about 0.73. Totally 84 Genotypes and 74 alleles were detected in all materials by 10 polymorphic markers, with the range from 4 to 12 and from 3 to 10 for each polymorphic primer set, respectively. The genetic distance among 42 tea varieties varied from 0.074 to 0.667, averagely 0.363, suggesting that the materials used in the experiment possess a broad genetic variation. Based on the similarity coefficient about 0.55, all the tea varieties tested could be classified into 3 groups and most of them were in first group. The results of this study proved that the EST-SSR marker is very effective in evaluation of tea germplasms.

Objective: To explore effective ways of integrated clinical and basic studies., Methods: Try to sum up clinically basic laws of acupuncture from the study thinking of clinical and basic integration., Results: Some laws of clinical acupuncture were raised based on classical theories of the channel and a great of clinical experiences, in combination with basic studies of modern neuroanatomy and physiology., Conclusion: The clinical symmetry-corresponding law, the segmental innervation law, the central-axial law, the distal limb dominance law, the law of sallow-deep correlation and the law of local feedback confirm with classical theories of the channels and clinical experiences.

28 pairs of primers were designed according to the expressed sequence tags in Chinese cabbage. After testing on the annealing temperature and the concentration of primer, dNTP and MgCl2, a suitable PCR system was established. Under the condition of reaction system developed, primers designed specific to ESTs were screened against genomic DNA of inbreed line A from which the cDNA library was constructed. Among them, 18 pairs of primers showed the amplification. Then all the primers available in line A were subjected to PCR for DNAs from 17 cabbage varieties. Polymorphism was detected by electrophoresis with agarose gel, and 10 of 18 primer sets could reveal polymorphisms among cabbage varieties, which accounted for 55.6% of primers selected. To examine the transferability of EST markers developed in cabbage, all primers were further used for PCR-mediated amplification of genomic DNA from different varieties of rapeseeds. Of 28 pairs of primers, 24 were able to produce amplified product(s) and 18 showed polymorphisms, accounting for 85.7% and 64.3% of total primers respectively. All of 18 primer sets that amplified in cabbage also showed amplified products in rapeseed and 13 of them were polymorphic. Even amongst the 10 primer sets that were unable to amplify in cabbage, 6 pairs produced amplification and 5 could reveal the polymorphism in rapeseeds. Results obtained in the present paper proved that developing polymorphic markers based on EST could be feasible and this kind of marker would be transferable to closed related species.

The segregation mode of transgenes was investigated in the transgenic progenies of three rice varieties (lines) produced by Agrobacterium-mediated transformation. The transgenic lines all contained the Bacillus thuringiensis cry1Ab gene, under the control of a maize ubiquitin promoter, and linked in tandem with gusA and hpt genes. PCR analysis showed the transgenes cry1Ab and gusA co-segregated in all self and crossed progenies tested. Therefore, GUS bio-assay of leaf or endosperm tissues was used to monitor transgene segregation in the experiment. It was found that the ratio of positive to negative plants was significantly smaller than 3:1 in all heterozygous plants derived progenies, which implied the segregation biased from typical Mendelian mode for a single dominant gene. Less GUS positive plants, and consequently less homozygous transgenic lines than expected were recovered from the self progenies. Transgenic heterozygous plants (+/-) were crossed as female or male parent to conventional rice varieties (-/-), and the ratio of gusA positive (+/-) to negative (-/-) plants was investigated in test F1 population. When used as female parent, the segregation fit to 1:1, but significantly smaller than 1:1 when used as male parent. The seed-set of transgenic Nipponbare progeny was investigated individually for GUS positive and negative plants. It was found that the positive plants had an average seed-set of 64.5%, significantly lower than that of negative plants (77.9%). The biological and genetic basis of distorted segregation of transgenes was discussed and deduced on the basis of above results, and the authors are inclined to ascribe these phenomena to the poor competitive ability of pollens carrying transgenes.

Two transgenic rice (Oryza sativa L.) lines, KMD1 and KMD2 at the R4 generation, transformed with a synthetic cry1Ab gene from Bacillus thuringiensis Berliner, were first evaluated for stem borer resistance in the field during the rice growing season of 1998 in two areas of Zhejiang Province, China. Both KMD1 and KMD2 were highly resistant to the stem borers Chilo suppressalis (Walker) and Scirpophaga incertulas (Walker), and were completely undamaged during the whole rice growing season. In contrast, damage to the plants of the untransformed parental control (Xiushui 11) was in the form of deadhearts or whiteheads. Under natural infestation by the C. suppressalis, the damage to control plants reached a peak of 88.7% of plants and 20.1% of tillers encountered with deadhearts. Under artificial and natural infestation of neonate striped stem borers at the vegetative stage and booting stage, 100% of plants and 25.6% of tillers, 78.9% of plants and 15.6% of productive tillers among artificially infested control plants were observed with the symptom of deadhearts and whiteheads, respectively. Damage to the control plants from artificial infestation by the S. incertulas reached a peak of 97.0% of plants and 22.9% of tillers damaged. The field research indicated that both KMD1 and KMD2 show great potential for protecting rice from attack by these two stem borers.

Improved histochemical staining for GUS activity, PCR and Western blotting were used to analyse the progeny population of Bt rice crossed with conventional rice varieties. A total of 392 plants expressing Bt toxin protein were found in 394 GUS positive plants. The result demonstrated that cry1Ab gene closely inherited and expressed with reporter gene gus. GUS assays indicated that cry1Ab gene of Bt rice inherits as a single dorminant gene. Southern hybridization confirmed cry1Ab gene can inherit stabily in the progenies of Bt rice. Mendelian segregation of reporter gene gus was also observed in F2, BC1 and BC1F2 progenies, which indicated that cry1Ab gene inherits as a single dorminant gene in the progenies of Bt rice crossed with conventional rice varieties.

Multidrug Resistance Protein 1 (MRP1) transports diverse organic anionic conjugates and confers resistance to cytotoxic xenobiotics. The protein contains two nucleotide binding domains (NBDs) with features characteristic of members of the ATP-binding cassette superfamily and exhibits basal ATPase activity that can be stimulated by certain substrates. It is not known whether the two NBDs of MRP1 are functionally equivalent. To investigate this question, we have used a baculovirus dual expression vector encoding both halves of MRP1 to reconstitute an active transporter and have compared the ability of each NBD to be photoaffinity-labeled with 8-azido-[(32)P]ATP and to trap 8-azido-[(32)P]ADP in the presence of orthovanadate. We found that NBD1 was preferentially labeled with 8-azido-[(32)P]ATP, while trapping of 8-azido-[(32)P]ADP occurred predominantly at NBD2. Although trapping at NBD2 was dependent on co-expression of both halves of MRP1, binding of 8-azido-ATP by NBD1 remained detectable when the NH(2)-proximal half of MRP1 was expressed alone and when NBD1 was expressed as a soluble polypeptide. Mutation of the conserved Walker A lysine 684 or creation of an insertion mutation between Walker A and B motifs eliminated binding by NBD1 and all detectable trapping of 8-azido-ADP at NBD2. Both mutations decreased leukotriene C(4) (LTC(4)) transport by approximately 70%. Mutation of the NBD2 Walker A lysine 1333 eliminated trapping of 8-azido-ADP by NBD2 but, in contrast to the mutations in NBD1, essentially eliminated LTC(4) transport activity without affecting labeling of NBD1 with 8-azido-[(32)P]ATP.

The two chain "Insulin/Insulin-like Growth Factor-I" hybrids, Ins/IGF-I(8) and Ins/IGF-I(11), were obtained by means of enzymatic semisynthesis, using desoctapeptide insulin (DOI) and the octapeptide and undecapeptide chemically synthesized according to the sequence 22-29 and 22-32 of IGF-I respectively as the starting materials. Comparative studies of the molecules with insulin indicate that the hybrid molecules retain in vivo the full activity of insulin. So the replacement of B27Thr by Asn and B30Ala by Thr, and the exchange of the sequence orders of B25 and B26, B28 and B29, as well as the extension of tripeptide (Gly-Tyr-Gly) at the B30 do not affect insulin activity.

In view of the similarity of the charge distribution between fibrin A alpha 148-161 and A chain 149-157 of urokinase, the latter might compete with fibrin A alpha 148-161 when single chain pro-urokinase is converted to double chain urokinase. To test this, the stretch of urokinase A chain 135-157 was separated from the low molecular weight urokinase, a competitive binding between this stretch and fibrin to tPA kringle-2 was shown by radio-binding assay. The inhibition of the stretch on the fibrin stimulated activation of plasminogen was demonstrated in the caseinolytic system. The synthesized novapeptide urokinase A chain 149-157 (R-peptide) showed a significant inhibition on the activation of plasminogen in the presence of fibrin. By contrasting finely with R-peptide, a synthesized novapeptide in which Arg154 and Arg156 were replaced by Asp (D-peptide) did not show any inhibition effect on the fibrin stimulated activation of plasminogen by tPA. These results suggest that the positively charged residues in the stretch 149-157 of urokinase are crucial for the inhibition of fibrin binding with the kringle domain of urokinase.