ObjectiveTo evaluate the effect of aminoguanidine on expression of inducible nitric oxide synthase (iNOS) in neurons in the small intestinal nerve plexus of starved rats.MethodsNinety male SD rats weighing 230-270 g were randomly divided into 3 groups:group normal control (group C,n =10) ; group starvation (group S,n=40) and group starvation + aminoguanidine (group A,n =40).The animals were allowed free access to water but no food during starvation in S and A groups.In group A the animals were given aminoguanidine 150 mg·kg-1 ·d-1 intraperitoneally during starvation.Ten animals were sacrificed at 3,5,7 and 9 d of starvation respectively and intestine specimens were taken for determination of ratio of intestinal transit using dextran blue-2000 as indicator.Then the specimens of intestinal myenteric nerve plexus of ileum were collected and stained by histochemistry with nicotinamide-adenine dinucleotide phosphate-d for determination of iNOS expression.ResultsStarvation significantly reduced the small intestinal transit and increased iNOS expression in neurons in the myenteric nerve plexus of small intestine in proportion to days of starvation in group S compared with group C.Intraperitoneal aminoguanidine significantly attenuated the starvation-induced changes in intestinal transit and iNOS expression.ConclusionAminoguanidine can attenuate the up-regulation of the expression of iNOS in neurons in the myenteric nerve plexus of small intestine induced by starvation and is helpful in promoting the intestinal transit in starved rats.

Objective Mutations of 13 CODIS (Combined DNA Index System) STR core loci in 532 cases of paternity testing were observed in confirming paternity, the mutation rate and the mutation type were studied. Methods 587 cases of paternity testing were routinely carried out using AmpFe STR Profiler Plus and Cofiler PCR Amplification Kits. When one or two STR exclusions were found, then HLA system and other blood groups were tested by molecular typing, and sixteen STR loci were genotyped by using PowerPlexl6 PCR Amplification Kit. If necessary, the genotyping of Y chromosome specific STR and HLA allelic sequencing were added. Results 1052 meiosis were observed among the 532 cases in confirmed paternity, 18 mutation events were found in 17 paternity cases. Single-locus mutation was observed in 16 cases, and mutation at two STR loci was observed in one case. The observed mutational loci include: D5S818, D3S1358, D16S539, CSFIPO, D21S11, D13S317, D7S820, vWA, D18S51 and FGA. The mutation rates for D18S51 and FGA loci were both 0.29% , which were the highest among the ten mutational loci. 11 events of paternal source mutations, 5 events of maternal source mutations and two events of indistinguishable mutations were observed in 18 STR mutational events. Conclusion When one or two STR exclusions were found in paternity testing, other more genetic markers must be detected as complement before making final conclusions.

Objective To explore the feasibility and reliability to build a sinoatrial node damage model in canine induced by formaldehyde wet dressing.Methods Twenty dogs were randomly divided into 4 groups(5 in each group) by means of random number table:2-hour,24-hour,1-week and 4-week groups after wet dressing.The sinoatrial node area of canine was damaged by wet dressing with 20% formaldehyde.Sinus node function was measured before,2 h after wet dressing and corresponding times of each group.Electrocardiogram(ECG) of body surface were recorded synchronically.Results Average wet dressing time was 6.2?2.6(3 to 12)min.Five dogs showed significantly decrease of heart rate(HR)(140?11 vs 89?6 beat/min,P

BACKGROUND: Umbilical cord blood (UCB) with limited karyocytes is mainly used in child patients. Recently, physicians have tried to mix two units of cord blood in the treatment of adults with hematological system diseases.OBJECTIVE: To monitor quantitatively the dynamic changes and the development rules of engraftment, chimera types and relative amount after allogeneic transplantation of mixed UCB from two units in adults with leukemia.DESIGN: Donors and the recipient were regarded as observational subjects in umbilical cord blood transplantation (UCBT). DNA extracted from blood samples of donors and the recipient before and after transplantation was considered as detecting samples. Short tandem repeat (STR) loci were as observational measures.SETTING: Key Laboratory of Immunology and Genetics of Institute of Transfusion Medicine of Shenzhen Blood Center.PARTICIPANT: A 43-year male patient with acute myeloid leukemia (AML), 75 kg, who was hospitalized at Shenzhen Hospital of Peking University, was enrolled in June 2005. The patient received two units of human leucocyte antigen (HLA), one locus mismatched unrelated UCBT (2.5×107 kg-1 karyocytes in UCB 1, and 1.53×107 kg-1 karyocytes in UCB 2) at month 6 after complete remission from first chemotherapy. UCB was collected from Guangzhou umbilical cord blood bank. The patient signed the informed consent.METHODS: The adult with AML received two units of HLA, one locus mismatched unrelated UCBT (2.5×107 kg-1 karyocytes in UCB 1, and 1.53×107 kg-1 karyocytes in UCB 2). Nine STR loci of the blood sample were determined before and after transplantation by quantitative technique of fluorescence labeling with multiplex polymerase chain reaction (MPCR), while the engraftment and chimera types were qualitatively evaluated by comparing differential loci between the recipient and the donors. The relative amount of two units of UCB was calculated in the patient after transplantation according to the differential gene peak areas of two donors with 377XL DNA sequencer after fluorescence scanning. The engraftment level and the development rules of donors' cells were analyzed quantitatively. In addition, the results were also compared with that of HLA loci distinct analysis for engraftment.MAIN OUTCOME MEASURES: After UCBT, transition process of nine STR loci of the recipient and two donors was observed, and engraftment was quantitatively and qualitatively described.RESULTS: Two units of UCB at day 15 after transplantation were engrafted simultaneously and revealed a complete chimera of the two. The relative amounts of UCB 1 and UCB 2 were 51.3% and 48.7%, respectively. Subsequently, UCB 1 went up to 70.0% and UCB 2 declined to 30.0% at day 30. However, only the genotype of UCB 1 was detected at day 52, and engraftment turned to a complete chimera of a single donor. The one with fewer karyocytes was rejected and the one with more karyocytes was engrafted for a long term.CONCLUSION: To detect quantitatively STR chimera with fluorescence labeling and MPCR can show precisely the engraftment level and the change of two units of UCB. It provides an accurate and reliable experimental basis for clinical UCB application and donor selection. It is proved that adult transplantation at the same time with mixed UCB from two units HLA one locus mismatched unrelated donors is feasible.

Objective:To find absorbable adhesives with suitable bonding properties for the absorbable polylactic acid root canal post. To test and compare the bond strengths of absorbable polylactic acid root canal post with three different adhesives. Methods:The absorbable polylactic acid root canal posts were used to restore the extracted teeth, using 3 different adhesives: cyanoacrylates, fibrin sealant and glass ionomer cement. The teeth were prepared into slices for micro-push-out test. The bond strength was statistically analyzed using ANOVA. The specimens were examined using microscope and the failure mode was divided into four categories:cohesive failure between absorbable polylactic acid root canal posts and adhesives, cohesive failure between dentin and adhesives, failure within the adhesives and failure within the absorbable polylactic acid root canal posts. Results:The bond strength of cyanoacrylates [(16. 83 ± 6. 97) MPa] and glass ionomer cement [(12. 10 ± 5. 09) MPa] were significantly higher than fibrin sealant [ ( 1 . 17 ± 0 . 50 ) MPa ] , P <0 . 001 . There was no significant difference between cyanoacrylates and glass ionomer cement (P =0. 156). In the group of cyanoacrylates, the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives was 25 . 0%, the cohe-sive failure between the dentin and the adhesives was 16. 7%, the failure within the adhesives was 33. 3%, and the failure within the absorbable polylactic acid root canal posts was 25 . 0%. In the group of fibrin sealant , the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives was 66 . 7%, the cohesive failure between the dentin and the adhesives was 22 . 2%, the failure within the ad-hesives was 11. 1%. In the group of glass ionomer cement, the cohesive failure between the absorbablepolylactic acid root canal posts and the adhesives was 87. 5%, the failure within the adhesives was 12. 5%. The major failure mode in fibrin sealant and glass ionomer cement was the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives. No major failure modes were found in the group of cyanoacrylates. Conclusion:The bond strength of fibrin sealant is low, which cannot meet the requirement of clinical use. The bond strengths of cyanoacrylates and glass ionomer cement are suitable for clinical use. The cyanoacrylates are a kind of absorbable adhesive which has suitable bonding proper-ties for the absorbable polylactic acid root canal post.

Objective To investigate the effects of resveratrol on the first and double oxygen-glucose deprivation (OGD) primary cortical neuron silent information regulator 1 (SIRT1), AMP-activated protein kinase (AMPK) activity and ATP content, and its possible neuroprotective mechanism. Methods Cortical neurons were taken from the embryos of 18-day Wistar rats. An in vitro repeated ischemia model was induced by the double OGD after the success of primary culture. Trypan blue stalning was used to detect the cel survival rate. Western blot was used to detect the SIRT1 and phospho-AMPK expression. Deacetylase fluorescence assay was used to detect the SIRT1 activity. Bioluminescence assay was used to detect the ATP content. Results Compared with the control group, resveratrol (0. 5 μmol/L) preconditioning significantly increased the survival rates after the single and double OGD (al P < 0. 001), ATP content (al P = 0. 004), SIRT1 activity (single: P = 0. 001; double: P = 0. 002), and the expression levels of SIRT1 (single: P = 0. 029; double: P = 0. 023) and phospho-AMPK (al P = 0. 001). Conclusions Resveratrol has the neuroprotective effect for the first and double OGD cortical neurons. Its mechanism may be associated with upregulating the SIRT1/AMPK signaling pathways and decreasing the energy requirements.

AIM: To identify human leukocyte antigen (HLA)-DRB11454 allele and HLA-DRB1 exon 3 sequence information in the Chinese population, which is significant for organ transplantation, cell transplantation, and human genetics.METHODS: Polymerase chain reaction sequence-based typing (PCR-SBT) was used to identify HLA-DRB1 alleles from 58 donor-recipient individuals who would undergo haemopoietic stem cell transplantation. Medium to high resolution polymerase chain reaction-reverse sequence specific oligonucleotide probe (PCR-RSSOP) was used to identify HLA-DRB1 alleles from 1 268 healthy donors from Guangdong province. The some ambiguous results of HLA-DRB114-associated alleles were confirmed by high resolution polymerase chain reaction-sequence-specific primer typing (PCR-SSP).RESULTS: HLA-DRB11403, 1406, 1410, 1412, 1418, 1425 and 1454 alleles were detected in 1 268 healthy donors.HLA-DRB11454 was confirmed in 8 ambiguous results of HLA-DRB11401/1434/1454 alleles, and HLA-DRB11454 was one of common alleles of HLA-DRB114 allele group in Guangdong population. HLA-DRB114 exon 3 sequence information was confirmed to be polymorphic in Chinese population.CONCLUSION: HLA-DRB11454 and exon 3 of DRB1 are confirmed to be polymorphic in Chinese population, further elucidating that HLA-DRB1 axon 3 sequence information is important for Han population and some minority groups.

Objective To explore the effect of high voltage alternating electric field therapy on fracture healing in rats. Methods An animal model of cortical bone defect was successfully established in the unilateral proximal tibia of 41 rats.They were then randomized into an experimental group (21 rats) and a control group (20 rats).The animals in the experimentsl group were given high voltage alternating electric field therapy for 30 minutes daily,while those in the control group only had sham exposure to the high voltage alternating electric field therapy.Rats in each group were sacrificed at days 7,14,21 and 28 post-operation.After blood collection,the operated tibias of the rats were resected with muscle and other soft tissue removed.Bone mineral density (BMD) of the callus at the fracture point was measured using dual energy X-ray absorptiometry (DEXA).The calluses were then cut down and fixed,decalcificated and sliced for histological observation.At the same time serum osteocalcin ( OC ) and bone alkaline phosphatase (RAP) were also detected by ABC-ELISA. Results The BMD examination showed that callus BMD in the experimental group was significantly higher than in the control group at days 14 and 28.Serum OC and RAP in the experimental group were also higher at days 14 day 21,and OC was still significantly higher in the experimental group at day 28.Histological examination showed that in the early stages of bone healing hematoma absorption and organization appeared earlier in the experimental group than in the control group,with remarkable osteoblast and chondroblast proliferation.While at the later stage,bone trabeculas connection and mineralization also appeared earlier in the experimental group than in the control group. Conclusions High voltage alternating electric field therapy may promote fracture healing by accelerating hematoma absorption and organization,elevating osteoblast proliferation and differentiation as well as promoting extracellular matrix secretion and mineralization.The underlying therapeutic mechanism might be related to the increased expression of OC and RAP caused by exposure to the high voltage alternating electric field.

BACKGROUND: The judgment of the engraftment of hematopoietic stem cells after transplantation mainly depends on various genetic labeling in vivo, which are different in sensitivity and effectiveness, thus a method with powerful differential ability, high sensitivity and not restricted by sex is to be established.OBJECTIVE: To observe the DNA genetic loci of short tandem repeat in the blood samples of both donors and recipients before allo-hematopoietic stem cell transplantation and those of recipients at different time points after transplantation.DESIGN: An observation measurement.SETTING: Laboratory of Immunogenetics, Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center.PARTICIPANTS: Blood samples of 18 pairs of donors and recipients, who were successfully matched and accepted hematopoietic stem cell transplantation, were selected from the Laboratory of Immunogenetics, Shenzhen Institute of Transfusion Medicine, Shenzhen Blood Center from February 2004 to December 2005. Among the 18 patients, there were 10 males and 8 females, with a mean age of 35 years old, including 6 cases of them were donated by relatives with blood relationship, and 12 cases by volunteers without blood relationship. Informed consents were obtained from all the participants.METHODS: The blood samples of both donors and recipients before transplantation and the blood samples of recipients after transplantation were collected, and the fluorescence labeling short tandem repeat technique was used to detect the 15 loci for short tandem repeat and Amelogenin sex locus, so that the differential loci between the donor and recipient could be screened. The engraftment and dynamic changes of the short tandem repeat genes of the donors in the recipients after transplantation were observed, the times for the earliest occurrences of short tandem repeat genes of the donors and the complete chimerism were recorded.MAIN OUTCOME MEASURES: ① Differential genes between the donors and recipients before transplantation;②Times for the earliest occurrences of short tandem repeat genes of the donors and the complete chimerism.RESULTS: All the 18 pairs of donors and recipients were involved in the final analysis of results. Satisfactory results of the typing at the 15 loci for short tandem repeat and 1 sex locus in the 18 pairs of samples of both donors and recipients before transplantation and the sample of the recipients after transplantation respectively. Averagely 12.4 (8-15) differential loci for short tandem repeat could be distinguished between the donors and recipients. ②After transplantation, short tandem repeat genes could be detected the earliest at 8 (5-14) days averagely, It took 14 (9-23) days averagely for short tandem repeat loci to convert from recipient type completely into donor type, and the engraftment converted from the recipient chimerism types completely into the donor types.CONCLUSION: The fluorescence labeling compound amplification of short tandem repeat technique can precisely measure the number of PCR products, describe the engraftment of hematopoietic stem cells and the whole process of development. It can also provide accurate and timely information for the early judgement of engraftment, predicting failure of transplantation and controlling recurrence.

To study the chemical constituents of Bauhinia glauca subsp. pernervosa, eleven phenolic acids were isolated from a 95% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, MCI, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, 2D NMR, and HR-ESI-MS, these compounds were identified as isopropyl O-beta-(6'-O-galloyl)-glucopyranoside (1), ethyl O-beta-(6'-O-galloyl)-glucopyranoside (2), 3, 4, 5-trimethoxyphenyl-(6'-O-galloyl)-O-beta-D-glucopyranoside (3), 3, 4, 5-trimethoxyphenyl-beta-D-glucopyranoside (4), gallic acid (5), methyl gallate (6), ethyl gallate (7), protocatechuic acid (8), 3, 5-dimethoxy-4-hydroxybenzoic acid (9), erigeside C (10) and glucosyringic acid (11). Among them, compound 1 is a new polyhydroxyl compound; compounds 2, 10, and 11 were isolated from the genus Bauhinia for the first time, and the other compounds were isolated from the plant for the first time. Compounds 6 and 8 showed significant protein tyrosine phosphatase1B (PTP1B) inhibitory activity in vitro with the IC50 values of 72.3 and 54.1 micromol x L(-1), respectively.