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Biomedical subjects

H Xin

Publications and source records attributed to H Xin.

At least 55 records · Page 3Linked to original sources

Assessing swine thermal comfort by image analysis of postural behaviors.

Postural behavior is an integral response of animals to complex environmental factors. Huddling, nearly contacting one another on the side, and spreading are common postural behaviors of group-housed animals undergoing cold, comfortable, and warm/hot sensations, respectively. These postural patterns have been routinely used by animal caretakers to assess thermal comfort of the animals and to make according adjustment on the environmental settings or management schemes. This manual adjustment approach, however, has the inherent limitations of daily discontinuity and inconsistency between caretakers in interpretation of the animal comfort behavior. The goal of this project was to explore a novel, automated image analysis system that would assess the thermal comfort of swine and make proper environmental adjustments to enhance animal wellbeing and production efficiency. This paper describes the progress and on-going work toward the achievement of our proposed goal. The feasibility of classifying the thermal comfort state of young pigs by neural network (NN) analysis of their postural images was first examined. It included exploration of using certain feature selections of the postural behavioral images as the input to a three-layer NN that was trained to classify the corresponding thermal comfort state as being cold, comfortable, or warm. The image feature selections, a critical step for the classification, examined in this study included Fourier coefficient (FC), moment (M), perimeter and area (P&A), and combination of M and P&A of the processed binary postural images. The result was positive, with the combination of M and P&A as the input feature to the NN yielding the highest correct classification rate. Subsequent work included the development of hardware and computational algorithms that enable automatic image segmentation, motion detection, and the selection of the behavioral images suitable for use in the classification. Work is in progress to quantify the relationships of postural behavior and physiological responses of pigs using thermographs. The results are expected to facilitate objective training of NN, hence improving the accuracy of the postural image-based assessment of the thermal comfort state. Work is also in progress to implement the analysis and assessment algorithms into computer codes for real-time application.

Animal Welfare↗

[The cytologic ultrastructure and cytochemical features of human dendritic cells from peripheral blood].

OBJECTIVE: To observe the cytomorphological, cellular ultrastructure and cytochemical features of the dendritic cell (DC) enriched populations in vitro. METHODS: Fresh peripheral blood from healthy volunteers was collected and mononuclear cells (PBMC) were isolated on lymphocyte separating solution. The PBMC were cultured in mediums containing rhGM-CSF, rhIL-4 and rhTNF-alpha 9-12 days to generate DC-enriched populations, the morphological features and numbers of DC were identified and counted respectively by light microscope (LM) and confirmed by immunofluorescence assay (IFA), the ultrastructure characteristics of the DC populations were observed by transmission electron microscope, and the cytochemical features of them were also examined. RESULTS: The percentage of DCs was 70%-80% in the DC-enriched populations as confirmed by IFA using mABXII. Transmission electron microscopy revealed that the DC populations contained two subsets of DC, one was considered as precursor cell derived DC(PC-DC), the cell surface of which had many irregular dendrites with a irregular nucleus, small nucleoli, numerous mitochondria, but scanty lysosomes, ribosome, rough endoplasmic reticulum and free polysomes. Cytochemical staining showed the DCs were both peroxidase (POX) and non-specific esterase (NSE) negative. The other was indentified as monocyte-derived DC(MO-DC), the features of which included: irregular shape, many ruffles and some rough lames on cell surfaces, nephritic or horseshoe nuclei with a thick rim of heterochromatin, numerous mitochondria, much more lysosomes, ribosomes, rough endoplasmic reticulum and Golgiosome. The cytochemical reactions indicated that most of them had POX-positive granules, and were NSE slightly positive. CONCLUSION: Cytokines in vitro induced DCs from human peripheral blood are heterogeneous.

Blood↗

[A study on formation of N-(nitrosomethyl) urea in experimental pig stomach gavaged with fish sauce].

OBJECTIVE: To study formation of N-(nitrosomethyl) urea (NMU), which is categorized as N-nitrosamides -- a sort of strong chemical carcinogen, by perfusion with fish sauce via pig stomach fistula, which is used daily as cooking flavor by local residents in Changle County, Fujian Province, a highly-prevalent area for gastric cancer. METHODS: Fistulization was performed in fasting experimental pigs' stomach, and their gastric juice was suctioned 30 minutes after perfusion with fish sauce and sodium nitrite via fistula. Gastric juice specimen was purified and concentrated and determined for NMU with high performance liquid chromatography-photolysis pyrolysis-thermal energy analyzer. RESULTS: NMU was detected in the pig stomach at pH 1 - 2, where NMU formed. Formation of NMU depended on the amount of nitrite added in a dose-dependent relationship. Level of NMU in gastric juice reached 25.4 and 7.97 micromol/L, respectively, when 3.48 and 0.87 mmol of sodium nitrite were fed into the stomach. No NMU could be detected, as 0.22 mmol of sodium nitrite fed. CONCLUSION: Under the condition of pH 1 - 2 and presence of sodium nitrite, NMU was synthesized in the stomachs of experimental pigs fed with fish sauce via gastric fistula. It is postulated that N-nitrosamides, such as NMU could be formed endogenously in the stomachs of local residents who consume fish sauce often and have a higher exposure to nitrite in their stomachs.

Animals↗

[Different protein forms of purple membrane produced by Halobacterium halobium under various nutritional condition].

The extremely halophilic bacterium, purple membrane producing Halobacterium halobium strain R1 were cultured in four liquid media which consist of different constituents. After the purple membrane is harvested by ultracentrification and purified by sucrose density gradient ultracentrification, and run on SDS-PAGE, the result of Coomassie blue staining showed different bR protein forms: three bands with molecule weight ranging from 26.0 to 27.5 kD in peptone medium, while only one band with MW 26.0 kD in other three media-complex, synthetic and artificial seawater media, which corresponded to the mature form in peptone medium. The result of Western blotting not only confirmed the different protein forms of Coomassie blue staining in the four media, but also gave additional bands that Coomassie blue can not detect due to its lower sensitivity: 1) in the complex, synthetic and artificial seawater media, a faint 28.0 kD band existed, while in the peptone medium it did not exist; and 2) in the four media, it appeared that a faint 23.0-24.0 kD band exited. The different bR protein forms of purple membrane which result from the difference of nutritional constituents in these four media might be caused by different precursor processing enzymes or different activities of precursor processing enzyme(s) under these different nutritional conditions.

Culture Media↗

Enhanced high-level expression of soluble 1-aminocyclopropane-1-carboxylase synthase and rapid purification by expanded-bed adsorption.

1-Aminocyclopropane-1-carboxylate (ACC) synthase is a key enzyme regulating the biosynthesis of the plant hormone ethylene. Expression of ACC synthase in Escherichia coli can result in the production of a large proportion of the enzyme in the form of insoluble aggregates (inclusion bodies). We investigated the effect on the soluble expression in E. coli of tomato and zucchini ACC synthases, by manipulation of the induction conditions, changing the vector, and deletions in the amino acid sequence. Manipulation of the induction conditions did not influence the soluble expression; however, soluble expression increased significantly when the enzyme was cloned into vector pET11d, in comparison to the other vector used, pET30a. It was also found that when ACC synthase with a portion of the C-terminus deleted was inserted into pET11d, the soluble expression was further enhanced in comparison to that of the full length. Structural and functional analysis of ACC synthase requires the purification of milligram quantities of protein to homogeneity. The development of a faster and simpler protocol for the purification of ACC synthase is highly desirable due to the extreme lability of the enzyme. C-terminal truncated tomato ACC synthase was overexpressed in E. coli pET11d and purified by expanded-bed adsorption and hydroxylapatite FPLC. This improved two-step purification protocol allows for rapid, high-level purification with a significantly improved yield in comparison to the multistage purification it replaces. 15.7 mg of highly purified tomato ACC synthase del-1 were obtained from 2 L of cells in comparison to 2 mg from 10 L using a multistage purification. This represents a 40-fold improvement in yield. Antibodies were raised against C-terminal deleted ACC synthase. The antibodies were purified by epitope-specific affinity chromatography and used to assess the identity and purity of the C-terminal-deleted tomato ACC synthase purified by expanded-bed adsorption.

Chromatography↗

Sonographically guided microwave coagulation treatment of liver cancer: an experimental and clinical study.

OBJECTIVE: Percutaneous microwave coagulation was performed with a modified system in animal experiments and in a clinical study to evaluate this technique as a treatment option for liver cancer. SUBJECTS AND METHODS: As an in vitro study, a microwave electrode was inserted 5-6 cm into separated egg white, homogenate of pig liver, and pig liver, with different power outputs and different lengths of inner conductors. In the animal experiment, the sonographically guided coagulation was performed percutaneously nine times and at laparotomy 43 times on 17 adult dogs. The thermal needles were placed parallel to and 5 mm, 8-12 mm, and 15 mm from the electrode. Clinically, 41 patients with hepatocellular carcinoma and 10 patients with hepatic metastases were treated with a 60-W microwave emission for 240-300 sec. RESULTS: Microwave coagulation using the modified system at 60 W for 300 sec produced a necrosis volume of 3.7 x 2.6 x 2.6 cm. The coagulated volume was elliptic when the exposed inner conductor of the electrode was 27 mm. The temperature at the periphery was 62.0 +/- 5.8 degrees C. During a mean follow-up period of 23 months, in 41 patients with hepatocellular carcinoma, 79% (46/58) of lesions became smaller, and the intratumoral blood flow disappeared in 89% (47/53). All tumors showed decreased density on unenhanced CT, and 84% (32/38) of tumors showed no enhancement on contrast-enhanced CT. In 21 patients with an elevated alpha-fetoprotein level, the level decreased in all 21 and was normalized in 17. A second biopsy on 19 patients showed complete destruction of tumor in 18. In 10 patients with hepatic metastases, the mean follow-up period was 13 months. Shrinkage of lesions occurred in 84% (21/25), and the blood flow inside the tumor disappeared in 75% (12/16) of lesions. Seventy-three percent (8/11) of the nodules showed no enhancement. A second biopsy on six patients showed complete necrosis in five. CONCLUSION: Sonographically guided microwave coagulation performed with this modified system was an effective and safe treatment for liver cancer.

Adult↗

[Effect of calcium overload and platelet activating factor on calcitonin gene related peptide release from lung tissue].

The level of calcitonin gene related peptide (CGRP) in plasma and pulmonary lavage fluid was tested by radio-immunoassay on intestinal ischemia/reperfusion (I/R) injured rat. The results showed that plasma CGRP after I/R was 161.3% more than control, and 117.8% when pre-treated with platelet activating factor (PAF) receptor antagonist SR27417A before injury but no obvious changes were observed in the sham group. Meanwhile, CGRP was reduced by 17.4% in pulmonary lavage fluid in I/R animal and elevated by 97.9% in SR27417A pre-treated animal, compared with normal control. Furthermore, it was demonstrated that CGRP release from lung tissue was enhanced concentration-dependently by calcium ionophores A23187 or PAF, as a result of blockage by phospholipase A2 inhibitor p-BPB or SR27417A respectively. In vitro experiment, CGRP in pulmonary lavage fluid was 144.6% more than control when pre-treated with SR27417A and 114.6% less than that treated with A23187 only, even 14.8% less than control. Much the same, CGRP release was 13.7% less than that treated with PAF and was 55.6% more than control when pre-treated with SR27417A only. All these results indicated that (1) CGRP was significantly increased in plasma but without obvious changes in pulmonary lavage fluid following I/R injury; (2) lung tissue CGRP release may be related with Ca2+ increase in cells or phospholipase A2 activation as well as PAF increase; (3) SR27417A was a kind of strong reagent, which may promote CGRP release in vivo and in vitro, but block the effect of PAF on CGRP release.

Animals↗

Expression of bovine mitochondrial elongation factor Ts in Escherichia coli and characterization of the heterologous complex formed with prokaryotic elongation factor Tu.

When bovine mitochondrial elongation factor Ts (EF-Ts(mt)) is expressed in Escherichia coli, it forms a tightly associated complex with E. coli EF-Tu (EF-Tu(Eco) x Ts(mt)). This complex is active in poly(U)-directed polymerization and this activity is inhibited by kirromycin. The EF-Tu(Eco) x Ts(mt) complex does not bind guanine nucleotides detectably and is not dissociated to a significant extent by either GDP or GTP. A portion of the EF-Tu(Eco) x Ts(mt) complex can be dissociated by aa-tRNA in the presence of GTP. The heterologous complex cannot be dissociated completely in the presence of either the 8 M urea or 8 M guanidine hydrochloride, suggesting that EF-Ts(mt) has an unusually tight interaction with E. coli EF-Tu. The EF-Tu(Eco) x Ts(mt) complex can be dissociated by denaturation using 2 M guanidine thiocyanate. Free EF-Ts(mt) can then be purified and renatured. The refolded EF-Ts(mt) is active in stimulating the activity of expressed mitochondrial EF-Tu (EF-Tu(mt)) in poly(U)-directed polymerization. Almost all the EF-Ts(mt) molecules appear to refold into a conformation which can interact with EF-Tu(mt). Protease mapping of EF-Ts(mt) indicates that the first 54 residues fold into an independent domain. Analysis of deletion derivatives of EF-Ts(mt) indicates that extensive regions of this factor are required for its tight interaction with EF-Tu.

Animals↗

[Experimental studies on effects of zinc and germanium on immune function and anti-oxidation in mice].

Zinc and germanium concentrations in serum, liver and muscle of mice, T-lymphocyte subgroup proportion, serum superoxide dismutase (SOD) activity and malonodialdehyde (MDA) were determined to study whether there exist synergism or antagonism between zinc and germanium. Results showed there existed, to certain extent, competitive effects of serum zinc and germanium in mice. When concentration of serum zinc increased, that of germanium decreased, or vise versa. There existed certain relationship between zinc and germanium concentrations in serum and those in muscle and liver of mice, and between those and CD3 count and SOD activity. Immune function in mice was influenced and their T-lymphocyte subgroup proportion changed with changes in serum zinc and germanium concentrations. With increased germanium and decreased zinc serum concentrations, CD3 and CD1 counts decreased, CD8 count unchanged, and the ratio of CD1 to CD3 decreased, which showed certain antagonist effects of them as they operated together. But, no antagonism was observed in their anti-oxidant effects, SOD activity increased to varied extent, and the level of MDA decreased.

Adjuvants, Immunologic↗

[A simplified theory of protrusive balanced occlusion in complete denture and its application].

Based the study of the theory of protrusive balanced occlusion in complete denture construction and clinical experience the author established a new simplified theory named "the three principle factors and three laws for protrusive balanced occlusion in complete denture" to guide us to arrange teeth for balanced occlusion in complete denture construction. Based this theory the phenomena of protrusive occlusal disharmony appeared in the process of teeth arrangement and denture insertion can be corrected easily.

Dental Occlusion, Balanced↗

Cloning and expression of mitochondrial translational elongation factor Ts from bovine and human liver.

The sequences of the cDNAs for the mitochondrial translational elongation factor Ts (EF-Tsmt) from bovine and human liver have been obtained. The deduced amino acid sequence of bovine liver EF-Tsmt is 338 residues in length and includes a 55-amino acid signal peptide and a mature protein of 283 residues. The sequence of the mature form of bovine EF-Tsmt is 91% identical to that of human EF-Tsmt and 29% identical to Escherichia coli EF-Ts. Southern analysis indicates that there are two genes for EF-Tsmt in bovine liver chromosomal DNA. A 224-base pair intron is located near the 5'-end of at least one of these genes. Northern analysis using a human multiple tissue blot indicates that EF-Tsmt is expressed in all tissues, with the highest levels of expression in skeletal muscle, liver, and kidney. Both the mature and precursor forms of bovine liver EF-Tsmt have been expressed in E. coli as histidine-tagged proteins. The mature form of EF-Tsmt forms a complex with E. coli elongation factor Tu. This complex is active in poly(U)-directed polymerization of phenylalanine. The precursor form is expressed as a 42-kDa protein, which is rapidly degraded in the cell.

Amino Acid Sequence↗

The covalently closed duplex form of the hepadnavirus genome exists in situ as a heterogeneous population of viral minichromosomes.

Replication of hepadnaviruses requires a persistent population of covalently closed circular (CCC) DNA molecules in the nucleus of the infected cell. It is widely accepted that the vital role of this molecule is to be the sole DNA template for the synthesis by RNA polymerase II of all viral transcripts throughout the infection process. Since the transcriptional activity of eukaryotic nuclear DNA is considered to be determined in part by its specific organization as chromatin, the nucleoprotein disposition of the hepadnavirus CCC DNA was investigated. These studies were undertaken on the duck hepatitis B virus (DHBV) CCC DNA present in the liver cell nuclei of DHBV-infected ducks. The organization and protein associations of the DHBV CCC DNA in situ were inferred from sedimentation, micrococcal nuclease digestion, and DNA superhelicity analyses. These three lines of investigation demonstrate that the DHBV CCC DNA is stably associated with proteins in the nuclei of infected liver cells. Moreover, they provide compelling evidence that the viral nucleoprotein complex is indeed a minichromosome composed of classical nucleosomes but in arrays that are atypical for chromatin. When the DHBV chromatin is digested with micrococcal nuclease, a ladder of viral DNA fragments that exhibits a 150-bp repeat is produced. This profile for the viral chromatin is obtained from the same nuclei in which the duck chromatin shows the standard 200-bp ladder. The superhelicity of the DHBV CCC DNA ranges from 0 to 20 negative supertwists per molecule, with all possible 21 topoisomers present in each DNA preparation. The 21 topoisomers of DHBV CCC DNA are inferred to derive from an identically diverse array of viral minichromosomes. In the DHBV minichromosomes composed of 20 nucleosomes, 96.7% of the viral DNA is calculated to be compacted into these chromatin subunits spaced on average by 5 bp of linker DNA; other minichromosomes contain fewer nucleosomes and proportionately more linker DNA. Two major subpopulations of DHBV minichromosomes are detected with comparable prevalence. The two groups correspond to minichromosomes which contain essentially a full or half complement of nucleosomes. The functional significance of this minichromosome diversity is unknown but is suggestive of transcriptional regulation of the viral DNA template.

Animals↗

Comparative mutagenicity of plant-activated aromatic amines using Salmonella strains with different acetyltransferase activities.

Plant systems can activate aromatic amines into mutagens. In the plant cell/microbe coincubation assay, we earlier demonstrated that 2-aminofluorene and m-phenylenediamine were activated by plant cells into mutagens with reversion at the hisD3052 allele in Salmonella typhimurium strain TA98 as the genetic endpoint. New derivatives have been developed which possess elevated levels of acetyl-CoA:N-hydroxyarylamine O-acetyltransferase which are very sensitive to N-hydroxylated amines [Watanabe et al., 1990: Mutat Res 234:337-348]. The objectives of this research were to examine Salmonella strains with different acetyltransferase activities in the plant cell/microbe coincubation assay with a series of structurally related aromatic amines. The hypothesis tested was whether and to what degree a plant-activated metabolite of these aromatic amines could serve as a substrate for bacterial O-acetyltransferase and induce mutation in Salmonella. Every aromatic amine examined was activated by plant cells with YG1024 (the strain with elevated O-acetyltransferase activity) as the genetic indicator organism. The rank order of the mutagenic responses of YG1024 to the plant-activated aromatic amines was 2-aminofluorene > benzidine > m-phenylenediamine > 4-aminobiphenyl > 2,4-diaminotoluene > 2-naphthylamine. This rank order was expressed by TA98 but to a much lower degree. There was a very slight mutagenic response observed in TA98/1,8-DNP6 (a strain lacking O-acetyltransferase activity) with the plant-activated metabolites of benzidine and 4-aminobiphenyl. We conclude that the plant-activated aromatic amines are substrates for bacterial O-acetyltransferases.

2-Naphthylamine↗

Feed and water consumption, growth, and mortality of male broilers.

Daily feed use, water use, body weight, and mortality of Cobb x Cobb male broilers over 8-wk growout periods were measured for 10 consecutive growouts in four commercial-scale broiler houses (121.0 x 12.1 m each). Polynomial equations were developed to relate bird age to body weight, daily feed and water use, cumulative weekly feed and water use, and cumulative mortality. Weekly feed conversion was derived from growth and feed use data and was depicted by a third-order polynomial equation. Dead bird weight was calculated using mortality and body weight of the broilers and related to bird age with three polynomial equations over the growth period. Total dead bird weight averaged 76 kg per 1,000 birds placed, of which 10 kg or 13% occurred during the first 5 wk and the remaining 66 kg or 87% occurred during the last 3 wk of the growout periods. Results of this study provide a realistic data base for mathematical modeling of production responses and a guideline for management planning in commercial male broiler operation.

Animals↗