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Biomedical subjects

L Jin

Publications and source records attributed to L Jin.

At least 127 records · Page 7Linked to original sources

[Association of single nucleotide polymorphisms in code region of beta2-adrenoceptor gene with hypertension in Chinese population].

OBJECTIVE: To detect single nucleotide polymorphisms(SNPs) existing in code region of beta(2)-adrenoceptor(2-AR) gene and to investigate association of the identified SNPs with essential hypertension in Chinese Han population. METHODS: Beta(2)-AR gene was sequenced with fluorescent labelling automatic sequencing method in unrelated Chinese Han population from Dabie Mountain in Anhui Province. Genotype of the SNPs were typed with PCR-RFLP method. RESULTS: Two SNPs were identified in length of 774bp, at position + 1053 with G-->C substitution and + 1239 with A-->G substitution respectively. The frequency of genotype of the two SNPs complied well with the Hardy-Weinberg equilibrium in normal group. Distribution of genotype AA, GA, GG of the SNPs at locus + 1239 in hypertension group was significantly different from that in normal group (chi(2) = 6.70, df = 2, P < 0.05). No significant difference was observed in distribution of genotypes of the SNPs at locus + 1053 between the two groups. CONCLUSION: These results indicate that the SNPs at locus + 1239 of beta(2)-AR gene is associated with EH. The SNPs at position + 1053 was not linked to hypertension.

Aged↗

Allogeneic bone nail for articular fractures.

OBJECTIVE: To investigate the effect of allogeneic bone nail on articular fractures. METHODS: Ninety-one patients with cancellous bone fracture involved in various joints were fixed with the bone nail or the bone screw. No immunosuppressive agent was used in this series. The average period of follow-up wa s 18 months. RESULTS: All patients except one healed without displacement an d the healing time was within 1-3 months. Joint function was partially limited in 6 patients and others were functioning well. CONCLUSIONS: Allogeneic bone nail possesses a good biological compatibility and intensity, thus provides a rigid fixation. Being completely absorbed and ossificated, thus obviating the need for secondary operation, is a suitable internal fixation material for the treatment of articular fractures.

Acetabulum↗

The influence of human single chain inteleukin-12 gene transduction on the biological behavior of hepatoma 7721 cells.

OBJECTIVE: To investigate the anti-tumor effects of human single chain interleukin-12 (hscIL-12). METHOD: pcDNA/hscIL-12 recombinant was transfected into human hepatic carcinoma cells (7721 cells) by lipofectin method. The 7721/hscIL-12 cells which secrete hscIL-12 stably, were obtained via G418 selection, and in vitro the influence of hscIL-12 gene transduction on the growth of tumor cells was evaluated by cell cycle analysis. In vivo, genetically engineered 7721 cells (7721/hscIL-12, 7721/pcDNA) and parental cells were implanted into BALB/c nude mice, respectively. 7721/pcDNA and 7721/hscIL-12 groups were divided into two sub-groups on day 8: one was administered with hPBL twice, 6 days at interval; the other was given equal volume of PBS. Mice were sacrificed on day 26, and spleens and tumors were taken out for histologic assay. RESULTS: hscIL-12 produced stably by 7721/hscIL-12 cells had bioactivity, and it was proved by Western blot, immunocytochemistry, and in situ hybridization. In vitro, compared with 7721 and 7721/pcDNA, the 7721/hscIL-12 grew much more slowly. FACS assay showed apparent G1 arrest of 7721/hscIL-12 cells. In animal experiment, on day 8 after inoculation, the tumors of 7721 and 7721/pcDNA group were up to 5 approximately 7 mm, while those of 7721/hscIL-12 group were 2 approximately 4 mm. When treated with hPBL, the tumor of 7721/hscIL-12 group disappeared completely. Histologically, the tumors from 7721/hscIL-12 without hPBL treatment had numerous lymphocyte infiltration, the tumor cells displayed depression looking, atrophy, focal necrosis and apoptosis, whereas the tumors of 7721 and 7721/pcDNA groups grew thrivingly. CONCLUSION: hscIL-12 transduced 7721 cells could induced significant antitumor immune response which resulted in tumor regression totally when the hPBL was inoculated, and also hscIL-12 has certain effects on mice immune system. These findings suggest that hscIL-12 and hscIL-12 gene therapy might have promising prospects in clinical application.

Animals↗

[Characteristics on ultraviolet absorption spectrum from difference in temperature and electron transition types of compounds].

The paper states a measuring method about Ultraviolet absorption spectrum from difference in temperature (UVSDT). The UVSDT of chemical compounds have been obtained with UV-240 ultraviolet spectrophotometer. The results show that the characteristic of UVSDT is related to electronic transition type of molecule. The long wave side shifts to long wave, when difference in temperature (delta t) accretes, peak high increases, bathochromic shift augments for UVSDT of n-sigma* electron transition. The UVSDT of n-pi* electron transition has two peak (one positive peak and one negative peak), when delta t accretes, the positive peak up increases, the negative peak down increases. The UVSDT of pi-pi* electron transition become negative peak, when delta t accretes, the negative peak down increases, the location of peak not moves.

Acetone↗

[Monte Carlo simulation of FCS in a laser gradient field].

Fluorescence correlation spectroscopy (FCS) is a powerful tool for probing biological process inside living cells. It measures fluorescence fluctuations of small number of molecules and derive information on molecular kinetics and reactions. We have developed a Monte Carlo model to simulate Browning motion of Rayleigh particles in a laser gradient field. The simulation reveals relations between laser field strength and measured parameters from FCS, such as diffusion coefficient and number density of the particles. The simulated results agree qualitatively to the experimental results obtained using fluorescent spheres. Empirical relations from the simulation are also discussed.

Computer Simulation↗

[Analysis on infrared spectrum of human body gallstone with comparative method].

The infrared spectra (IRS) are measured for chemical composition of series gallstones. The infrared spectra (IRS) of the gallstone are compared with IRS of standard compound sample. The typical IRS are divided into 6 genera in component analysis of gallstone according to the principal characteristic peak: cholesterol, calcium bilirubin, calcium stearate, calcium carbonate, hydroxyl carbonate phosphatic rock and mixture. The gallstones are correlatively divided into 6 genera of gallstones or 2 genera-cholesterol and calcium salt gallstones according to whether containing calcium salt.

Bilirubin↗

Atypical measles in a patient twice vaccinated against measles: transmission from an unvaccinated household contact.

Described are two cases within the same household that were involved in an outbreak of measles in Niterói, RJ. Measles diagnosis was confirmed serologically by specific IgM detection in Case 1 (classic measles) who was unvaccinated, and rising measles specific IgG in the absence of IgM in Case 2 (mild modified measles) who had a history of two vaccinations with measles-containing vaccines. Measles virus was detected by reverse transcriptase polymerase chain reaction (RT-PCR) in saliva samples from both cases. The nucleic acid amplified by RT-PCR was sequenced and showed identical measles sequence in the two cases. This study highlights the difficulty of diagnosing nonclassical measles infection on clinical and serological grounds, and the usefulness of PCR for viral RNA sequencing from noninvasive specimens for confirming epidemiologic links.

Adult↗

Amperometric ultramicrosensors for peroxynitrite detection and its application toward single myocardial cells.

The research studied the concentration variation of peroxynitrite anion (O=N-O-O-) released from cultured neonatal myocardial cells induced by ischemia/reperfusion and studied the protective effect of melatonin on the injury. For this purpose, amperometry peroxynitrite ultramicrosensors (UMS) were fabricated and constructed by electropolymerizing inorganic macromolecular film of tetraaminophthalocyanine manganese(II) and coating chemically with poly(4-vinylpyridine). Under optimum conditions, the UMS showed high selectivity and sensitivity to peroxynitrite determination with a calculated detection limit of 1.8 x 10(-8) mol/L (S/N of 3). The detection of peroxynitrite was based on electrocatalytic reduction of peroxynitrite. The mechanism of catalysis was also discussed. The UMS should be promising for in vivo measurement of peroxynitrite without interference or fouling. Peroxynitrite released from myocardial cells both in the ischemic period and in the reperfusion period was measured directly. This approach may lead to important information for myocardial cells on the mechanism of injury and prospective treatments of medicine such as melatonin.

Animals↗

Enhanced murine macrophage TNF receptor shedding by cytosine-guanine sequences in oligodeoxynucleotides.

The immunomodulatory role of unmethylated cytosine-guanine sequences (CpG) in bacterial DNA has been well documented. We have previously demonstrated that murine macrophage-like RAW 264.7 cells respond to CpG DNA with an increase in the proinflammatory cytokine, TNF-alpha, in both a dose-dependent and time-dependent manner. In addition, CpG DNA stimulates a significant, though delayed, secretion of the anti-inflammatory cytokine IL-10. Because TNF-alpha and TNFR (TNFRI and II) expression are tightly regulated responses, we hypothesized that CpG containing oligodeoxynucleotide (CpG ODN) would also affect TNFRI and II shedding. Using both murine peritoneal macrophages and RAW 264.7 cells, we demonstrated a significant, time-dependent increase in soluble TNFRI and TNFRII production with CpG ODN stimulation. RAW 264.7 cells treated with CpG ODN had a transient increase in membrane TNFRII expression, but not TNFRI. Both types of TNFR mRNA were also up-regulated by CpG ODN, and addition of the transcriptional inhibitor actinomycin D abrogated the effect of CpG ODN on TNFR mRNA and protein expression. Addition of anti-IL-10 and anti-TNF-alpha Abs did not change these results. The addition of plate-bound anti-TNF receptor Abs to this system increased the amount of bioactive TNF, implying that these receptors are acting as inhibitors of TNF activity. These results suggest that the de novo, non-IL-10- and non-TNF-alpha-dependent transcription, translation, and shedding of TNFRs are additional potential counterinflammatory effects of CpG DNA.

Adjuvants, Immunologic↗

Worldwide DNA sequence variation in a 10-kilobase noncoding region on human chromosome 22.

Human DNA sequence variation data are useful for studying the origin, evolution, and demographic history of modern humans and the mechanisms of maintenance of genetic variability in human populations, and for detecting linkage association of disease. Here, we report worldwide variation data from a approximately 10-kilobase noncoding autosomal region. We identified 75 variant sites in 64 humans (128 sequences) and 463 variant sites among the human, chimpanzee, and orangutan sequences. Statistical tests suggested that the region is selectively neutral. The average nucleotide diversity (pi) across the region was 0.088% among all of the human sequences obtained, 0.085% among African sequences, and 0.082% among non-African sequences, supporting the view of a low nucleotide diversity ( approximately 0.1%) in humans. The comparable pi value in non-Africans to that in Africans indicates no severe bottleneck during the evolution of modern non-Africans; however, the possibility of a mild bottleneck cannot be excluded because non-Africans showed considerably fewer variants than Africans. The present and two previous large data sets all show a strong excess of low frequency variants in comparison to that expected from an equilibrium population, indicating a relatively recent population expansion. The mutation rate was estimated to be 1.15 x 10(-9) per nucleotide per year. Estimates of the long-term effective population size N(e) by various statistical methods were similar to those in other studies. The age of the most recent common ancestor was estimated to be approximately 1.29 million years ago among all of the sequences obtained and approximately 634,000 years ago among the non-African sequences, providing the first evidence from a noncoding autosomal region for ancient human histories, even among non-Africans.

Animals↗

Conformational changes in serpins: II. The mechanism of activation of antithrombin by heparin.

Antithrombin, uniquely among plasma serpins acting as proteinase inhibitors in the control of the blood coagulation cascade, circulates in a relatively inactive form. Its activation by heparin, and specifically by a pentasaccharide core of heparin, has been shown to involve release of the peptide loop containing the reactive centre from partial insertion in the A sheet of the molecule. Here we compare the structures of the circulating inactive form of antithrombin with the activated structure in complex with heparin pentasaccharide. We show that the rearrangement of the reactive centre loop that occurs upon activation is part of a widespread conformational change involving a realignment of the two major domains of the molecule. We also examine natural mutants that possess high affinity for heparin pentasaccharide, and relate the kinetics of their interaction with heparin pentasaccharide to the structural transitions occuring in the activation process.

Amino Acid Sequence↗

Crystal structure of human parathyroid hormone 1-34 at 0.9-A resolution.

The N-terminal fragment 1-34 of parathyroid hormone (PTH), administered intermittently, results in increased bone formation in patients with osteoporosis. PTH and a related molecule, parathyroid hormone-related peptide (PTHrP), act on cells via a common PTH/PTHrP receptor. To define more precisely the ligand-receptor interactions, we have crystallized human PTH (hPTH)-(1-34) and determined the structure to 0.9-A resolution. hPTH-(1-34) crystallizes as a slightly bent, long helical dimer. Analysis reveals that the extended helical conformation of hPTH-(1-34) is the likely bioactive conformation. We have developed molecular models for the interaction of hPTH-(1-34) and hPTHrP-(1-34) with the PTH/PTHrP receptor. A receptor binding pocket for the N terminus of hPTH-(1-34) and a hydrophobic interface with the receptor for the C terminus of hPTH-(1-34) are proposed.

Amino Acid Sequence↗

Polynesian origins: insights from the Y chromosome.

The question surrounding the colonization of Polynesia has remained controversial. Two hypotheses, one postulating Taiwan as the putative homeland and the other asserting a Melanesian origin of the Polynesian people, have received considerable attention. In this work, we present haplotype data based on the distribution of 19 biallelic polymorphisms on the Y chromosome in a sample of 551 male individuals from 36 populations living in Southeast Asia, Taiwan, Micronesia, Melanesia, and Polynesia. Surprisingly, nearly none of the Taiwanese Y haplotypes were found in Micronesia and Polynesia. Likewise, a Melanesian-specific haplotype was not found among the Polynesians. However, all of the Polynesian, Micronesian, and Taiwanese haplotypes are present in the extant Southeast Asian populations. Evidently, the Y-chromosome data do not lend support to either of the prevailing hypotheses. Rather, we postulate that Southeast Asia provided a genetic source for two independent migrations, one toward Taiwan and the other toward Polynesia through island Southeast Asia.

Biological Evolution↗

A cis-proline to alanine mutant of E. coli aspartate transcarbamoylase: kinetic studies and three-dimensional crystal structures.

The only cis-proline residue in Escherichia coli aspartate transcarbamoylase has been replaced by alanine using site-specific mutagenesis. The Pro268-->Ala enzyme exhibits a 40-fold reduction in enzyme activity and decreased substrate affinity toward carbamoyl phosphate and aspartate compared to the corresponding values for the wild-type enzyme. The concentration of the bisubstrate analogue N-phosphonacetyl-L-aspartate (PALA) required to activate the mutant enzyme to the same extent as the wild-type enzyme is significantly increased. The heterotropic effects of ATP and CTP upon the Pro268-->Ala enzyme are also altered. Crystal structures of the Pro268-->Ala enzyme in both T- and R-states show that the cis-peptidyl linkage between Leu267 and Ala268 is maintained. However, the tertiary structure of both the catalytic and regulatory chains has been altered by the amino acid substitution, and the mobility of the active-site residues is increased for the R-state structure of Pro268-->Ala enzyme as comparison with the wild-type R-state structure. These structural changes are responsible for the loss of enzyme activity. Thus, Pro268 is required for the proper positioning of catalytically critical residues in the active site and is important for the formation of the high-activity high-affinity R-state of E. coli aspartate transcarbamoylase.

Alanine↗

Conformational correlation and coupled motion between residue A21 and B25 side chain observed in crystal structures of insulin mutants at position A21.

The C-terminal residue of the insulin A chain is invariant and kept as asparagine in all known insulin molecules from hagfish through birds to mammals. To get information on the role of this conserved residue, which is still unclear, the three-dimensional structures of four human insulin mutants, A21 Asn-->Gly, A21 Asn-->Asp, A21 Asn-->Ala, and A21 Asn-->Gln DesB30, were determined by X-ray crystallography. The four mutants crystallize separately into two kinds (rhombohedral and cubic) of crystals. In the refined structures, conformational correlation and coupled motion between the A chain C-terminal residue A21 and the B25 side chain was observed, in contrast to the nearly unchanged general structures as compared with the native insulin structures in their respective crystals. A detailed analysis suggests that residue A21 can affect insulin receptor binding by interaction with the B25 side chain and the B chain C-terminal segment to assist the B25 side chain rearranging into the 'active' conformation.

Crystallography, X-Ray↗

Expansion of genetically modified primary human hemopoietic cells using chemical inducers of dimerization.

The inability to deliver a therapeutic gene to a sufficient percentage of hematopoietic stem cells is the major obstacle to using gene therapy to treat blood disorders. Providing genetically corrected stem cells with a reversible growth advantage could solve this problem. To this end we have employed small synthetic molecules that can reversibly dimerize and activate fusion proteins which contain a growth factor receptor signaling domain. We have shown that the thrombopoietin receptor (mpl) signaling domain can be used in this system to expand transduced multipotential progenitor cells from mouse bone marrow. In the present study we tested a similar retroviral vector in human CD34-selected cord blood cells. Following transduction, cells cultured in the presence of the dimerizing molecule AP1903 expanded 13.8- to 186-fold relative to cells cultured in the absence of AP1903. The cell type that emerged in suspension culture was erythroid. Contrary to our results in the murine system, cell expansion was transient. Activation of mpl caused the disappearance of BFU-E followed by a transient increase in CFU-E. In contrast, mpl activation had no discernable effect on transduced myeloid progenitor cells. AP1903-mediated expansion was restricted to transduced cells, as demonstrated by immunohistochemical staining. These findings indicate that synthetic dimerizing molecules can be used to expand primary human hematopoietic cells. (Blood. 2000;95:430-436)

Animals↗