Biomedical subjects

B Pan

Publications and source records attributed to B Pan.

At least 37 records · Page 2Linked to original sources

PubMed1999

Solution structure of the single-strand break repair protein XRCC1 N-terminal domain.

XRCC1 functions in the repair of single-strand DNA breaks in mammalian cells and forms a repair complex with beta-Pol, ligase III and PARP. Here we describe the NMR solution structure of the XRCC1 N-terminal domain (XRCC1 NTD). The structural core is a beta-sandwich with beta-strands connected by loops, three helices and two short two-stranded beta-sheets at each connection side. We show, for the first time, that the XRCC1 NTD specifically binds single-strand break DNA (gapped and nicked). We also show that the XRCC1 NTD binds a gapped DNA-beta-Pol complex. The DNA binding and beta-Pol binding surfaces were mapped by NMR and found to be well suited for interaction with single-strand gap DNA containing a 90 degrees bend, and for simultaneously making contacts with the palm-thumb of beta-Pol in a ternary complex. The findings suggest a mechanism for preferential binding of the XRCC1 NTD to flexible single-strand break DNA.

Amino Acid Sequence↗

PubMed1999

The dimerization and topological specificity functions of MinE reside in a structurally autonomous C-terminal domain.

Correct placement of the division septum in Escherichia coli requires the co-ordinated action of three proteins, MinC, MinD and MinE. MinC and MinD interact to form a non-specific division inhibitor that blocks septation at all potential division sites. MinE is able to antagonize MinCD in a topologically sensitive manner, as it restricts MinCD activity to the unwanted division sites at the cell poles. Here, we show that the topological specificity function of MinE residues in a structurally autonomous, trypsin-resistant domain comprising residues 31-88. Nuclear magnetic resonance (NMR) and circular dichroic spectroscopy indicate that this domain includes both alpha and beta secondary structure, while analytical ultracentrifugation reveals that it also contains a region responsible for MinE homodimerization. While trypsin digestion indicates that the anti-MinCD domain of MinE (residues 1-22) does not form a tightly folded structural domain, NMR analysis of a peptide corresponding to MinE1-22 indicates that this region forms a nascent helix in which the peptide rapidly interconverts between disordered (random coil) and alpha-helical conformations. This suggests that the N-terminal region of MinE may be poised to adopt an alpha-helical conformation when it interacts with the target of its anti-MinCD activity, presumably MinD.

Amino Acid Sequence↗

PubMed1999

[Clinical application of 96 hours' rapid skin tissue expanding technique].

OBJECTIVE: To reduce expansion course, diminish complications and improve expanding effect in skin tissue expander treatment. METHODS: We have chosen 38 cases of skin tissue defects of small and moderate areas, on which we have carried out 96 hours' rapid skin tissue expanding technique by altering the time and the amount of saline injected. RESULTS: All skin tissue defects have been successfully repaired. Half a year later, the incisions were inconspicuous. CONCLUSION: 96 hours' rapid skin tissue expanding technique has the following advantages, it is simple and safe, economic and convenient, rapid and reliable. This technique tends to be mature. What is more, it is easy to be popularized. We regard it as an ideal rapid expansion technique.

Adolescent↗

PubMed1999

[The effect of combined endocrine therapy on prostate and testis].

OBJECTIVE: To study the pathological changes of the effect of combined endocrine therapy on normal prostate, benign prostatic hyperplasia, prostatic adenocarcinoma and testis. METHODS: 11 radical prostatectomy specimens, 3 core-needle biopsies of prostate and 3 testes obtained after at least 3 months of enatone-flutamide inhibition therapy were studied. Step-section was performed on radical specimens and average 16 sections per case were reviewed. PSA, PSAP and AE1/AE3 were immunostained on 14 cases of prostate cancer and a comparative study pre- and post-treatment was made. RESULTS: No residual tumor was recognized in 2 cases. Such characteristic changes were found in 9 cases as prominent acinar atrophy, decreased ratio of acini to stroma, stromal fibrosis, squamous metaplasia of carcinoma, cytoplasmic vaculation, nuclear shrinkage, and nucleolar shrinkage. No apparent change was discovered in 3 cases. Secretory epithelial atrophy and basal cell hyperplasia were the popular change in BPH and normal prostate. The expressions of both PSA and PSAP were markedly reduced in prostate carcinoma and nonneoplastic glands. Pathological downstaging of the tumor was not found statistically. Epithelial atrophy also existed in seminal vesicles and Leydig cells of the testes after hormone therapy. CONCLUSIONS: Combined endocrine therapy results in histologically distinctive changes that can be found in both nonneoplastic and neoplastic prostate tissue. However, the drugs can not eradicate prostate cancer completely. Testis atrophy is the direct action of the therapy.

Aged↗

PubMed1999

[The isolation of microbe producing biosurfactants].

Microorganisms capable of producing biosurfactants can be isolated by a series of steps including hydrocarbon enrichment culture, hemolytic activity assay on blood agar plates and oil displacement activity assay etc. Ten strains(including bacteria, yeasts and fungi) were isolated with higher surface activity from 1000 samples from soil, sludge, waste water etc. Two strains of bacteria produced rhamnolipids and trehaloselipids respectively. One strain of bacterium produced long chain unsaturated carboxylic acid and another produced azelaic acid. Two strains of yeasts produced lipopolysaccharides with excellent emulsification property.

Bacteria↗

PubMed1998

Crystal structure of an RNA octamer duplex r(CCCIUGGG)2 incorporating tandem I.U wobbles.

The crystal structure of the RNA octamer duplex r(CCCIUGGG)2has been elucidated at 2.5 A resolution. The crystals belong to the space group P21and have unit cell constants a = 33.44 A, b = 43.41 A, c = 49.39 A and beta = 104.7 degrees with three independent duplexes (duplexes 1-3) in the asymmetric unit. The structure was solved by the molecular replacement method and refined to an Rwork/Rfree of 0.185/0.243 using 3765 reflections between 8.0 and 2.5 A. This is the first report of an RNA crystal structure incorporating I.U wobbles and three molecules in the asymmetric unit. Duplex 1 displays a kink of 24 degrees between the mismatch sites, while duplexes 2 and 3 have two kinks each of 19 degrees and 27 degrees, and 24 degrees and 29 degrees, respectively, on either side of the tandem mismatches. At the I.U/U.I mismatch steps, duplex 1 has a twist angle of 33.9 degrees, close to the average for all base pair steps, but duplexes 2 and 3 are underwound, with twist angles of 24.4 degrees and 26.5 degrees, respectively. The tandem I.U wobbles show intrastrand purine-pyrimidine stacking but exhibit interstrand purine-purine stacking with the flanking C.G pairs. The three independent duplexes are stacked non-coaxially in a head-to-tail fashion to form infinite pseudo-continuous helical columns which form intercolumn hydrogen bonding interactions through the 2'-hydroxyl groups where the minor grooves come together.

Base Pairing↗

PubMed1998

Structure of a 16-mer RNA duplex r(GCAGACUUAAAUCUGC)2 with wobble C.A+ mismatches.

The crystal structure of a 16-mer, the longest known RNA duplex, has been determined at 2.5 A resolution. The hexadecamer r(GCAGACUUAAAUCUGC) contains isolated C.A/A.C mismatches with two hydrogen bonds. The two hydrogen bonds in the mismatches suggests that N1 of A is protonated even though the crystallization was done at neutral pH. Therefore, the C.A mismatch is a C.A+ wobble similar to the G.U wobble. The two C.A+ pairs are isolated by four Watson-Crick pairs and flanked by five Watson-Crick base-pairs on either sides. Kinks/bends of 20 degrees are observed at the wobble sites. The Watson-Crick base-pair A5.U26 on the 5'-side of the first C6.A27(+) wobble has a twist angle of 27 degrees compared to the 3'-side U7.A28 pair of 36 degrees. The twist angles are reversed (37 degrees and 26 degrees) in the second A11(+).C22 wobble because of the approximate dyad in the molecule, the flanking base-pair sequences are A.U pairs. The wobbles expand the major groove to 7.1 A/7.3 A. The duplexes form helical columns and are tightly packed around the 31-screw axis. The minor grooves of adjacent columns in juxtaposition interact through the O2' atoms and the anionic phosphate oxygen atoms.

Base Pair Mismatch↗

PubMed1998

[Primary epididymal tumors].

OBJECTIVE: To improve the diagnosis of primary epididymal tumors. METHOD: In 23 primary epididymal tumors (22 benign and 1 malignant), 14 were adenomatoid tumors, 6 were leiomyomas, 1 was multiple fibrous pseudomonas and 1 was sclerosing hemangioma. Epididymal adenocarcinoma was seen in a 57-year-old man. RESULT: Epididymal tumors might be easily misdiagnosed as epididymal tuberculosis, chronic epididymis and spermatocele. Apart from certain benign clinical characteristics, benign epididymal tumors could be diagnosed by B-ultrasonography and aspiration biopsy. Most malignant tumors of epididymis presented as rapidly growing scrotal masses and cured by the removal of tumors or the whole epididymis of the same side. 19 of 22 cases were free from recurrence. CONCLUSION: Malignant epididymal tumors should be treated as malignant tumors of testis and their prognosis is extremely poor.

Adenocarcinoma↗

PubMed1997

Secondary and tertiary structural changes in gamma delta resolvase: comparison of the wild-type enzyme, the I110R mutant, and the C-terminal DNA binding domain in solution.

gamma delta Resolvase is a site-specific DNA recombinase (M(r) 20.5 kDa) in Escherichia coli that shares homology with a family of bacterial resolvases and invertases. We have characterized the secondary and tertiary structural behavior of the cloned DNA binding domain (DBD) and a dimerization defective mutant in solution. Low-salt conditions were found to destabilize the tertiary structure of the DBD dramatically, with concomitant changes in the secondary structure that were localized near the hinge regions between the helices. The molten tertiary fold appears to contribute significantly to productive DNA interactions and supports a mechanism of DNA-induced folding of the tertiary structure, a process that enables the DBD to adapt in conformation for each of the three imperfect palindromic sites. At high salt concentrations, the monomeric I110R resolvase shows a minimal perturbation to the three helices of the DBD structure and changes in the linker segment in comparison to the cloned DBD containing the linker. Comparative analysis of the NMR spectra suggest that the I110R mutant contains a folded catalytic core of approximately 60 residues and that the segment from residues 100 to 149 are devoid of regular structure in the I110R resolvase. No increase in the helicity of the linker region of I110R resolvase occurs on binding DNA. These results support a subunit rotation model of strand exchange that involves the partial unfolding of the catalytic domains.

Ammonium Sulfate↗

PubMed1997

Crystal structure of d(GCGCGCG) with 5'-overhang G residues.

The crystal structure of the DNA heptamer d(GCGCGCG) has been solved at 1.65 A resolution by the molecular replacement method and refined to an R-value of 0.184 for 3598 reflections. The heptamer forms a Z-DNA d(CGCGCG)2 with 5'-overhang G residues instead of an A-DNA d(GCGCGC)2 with 3'-overhang G residues. The overhang G residues from parallel strands of two adjacent duplexes form a trans reverse Hoogsteen G x G basepair that stacks on the six Z-DNA basepairs to produce a pseudocontinuous helix. The reverse Hoogsteen G x G basepair is unusual in that the displacement of one G base relative to the other allows them to participate in a bifurcated (G1)N2 . . . N7(G8) and an enhanced (G8)C8-H . . . O6(G1) hydrogen bond, in addition to the two usual hydrogen bonds. The 5'-overhang G residues are anti and C2'-endo while the 3'-terminal G residues are syn and C2'-endo. The conformations of both G residues are different from the syn/C3'-endo for the guanosine in a standard Z-DNA. The two cobalt hexammine ions bind to the phosphate groups in both GpC and CpG steps in Z(I) and Z(II) conformations. The water structure motif is similar to the other Z-DNA structures.

Base Composition↗

PubMed1997

Chemical sensory deafferentation abolishes hypothalamic pituitary activation induced by noxious stimulation or electroacupuncture but only decreases that caused by immobilization stress. A c-fos study.

We have shown in previous c-fos studies that noxious stimulation or electroacupuncture in deeply anaesthetized rats activate the hypothalamic pituitary corticotrope axis in a specific way. C-fos expression was more pronounced in the arcuate than the paraventricular hypothalamic nuclei, and none occurred in the pituitary intermediate lobe. The absence of the usual autonomic responses to psychological stress, such as tachycardia or blood pressure elevation, suggested a specific action of the somatosensory input on the hypothalamic pituitary axis. To prove this hypothesis, c-fos expression was examined in the paraventricular, arcuate and other hypothalamic nuclei, the pituitary gland, and the A1 and A2 medullary catecholaminergic cell groups of animals deprived of nociceptive primary afferent input by neonatal capsaicin. After noxious stimulation or electroacupuncture, no c-fos enhancement occurred in any of those sites in capsaicin-treated animals, and there was no increased plasma release of adrenocorticotropic hormone. In contrast, the hypothalamic pituitary c-fos activation provoked by immobilization stress though markedly decreased, was not abolished by capsaicin, whereas plasma release of adrenocorticotropic hormone remained undiminished. These findings suggest that noxious stimulation or electroacupuncture act on the hypothalamic pituitary corticotrope axis through an exclusively physical effect depending on the noxious signal elicited in the somatosensory pathway. They also demonstrate the occurrence of a minor somatosensory physical component after forced immobilization, acting on the hypothalamic pituitary axis probably together with the prevalent component of emotional arousal elicited by this form of stress.

Adrenocorticotropic Hormone↗

PubMed1997

[Severe vascular injuries following total hip replacement].

Severe vascular injury following total hip replacement (THR) is very rare. It has an a incidence of 0.25% and poses a serious threat to the involved limb and the patient's life. Vascular structures affected include the external iliac artery and vein, femoral artery and vein. In 721 cases of THR 3 had vascular injuries. The first case sustained the injury of the external iliac vessel which was cut by the sharp edge of bone cement while removing the displaced prosthesis. The second had laceration of the iliac vein while a pneumatic drill was used in the anterion superior acetabulum, and the third had the cut injury of the femoral artery during resection of the capsular scar tissue. The understanding of the mechanisms and location of vascular injury during THR will help surgeons in early diagnosis and treatment of the injury.

Adult↗

PubMed1996

Activation of anterior lobe corticotrophs by electroacupuncture or noxious stimulation in the anaesthetized rat, as shown by colocalization of Fos protein with ACTH and beta-endorphin and increased hormone release.

A marked expression of the c-fos proto-oncogene has been recently reported in cells of the anterior lobe of the pituitary gland in rats subject to electroacupuncture or noxious thermal stimulation under pentobarbital anaesthesia. The present study was undertaken to identify the activated pituitary cells. Following both kinds of stimulation, most Fos-immunoreactive anterior lobe cells showed colocalization with adrenocorticotropic hormone or beta-endorphin immunoreactivity. No c-fos expression occurred in pituitary cells immunoreactive for growth hormone, prolactin, luteinizing hormone, or thyrotropin-stimulating hormone. A marked rise of adrenocorticotropic hormone and beta-endorphin concentrations occurred in plasma. In the hypothalamus, c-fos expression was increased in the mediobasal nuclei-namely, the arcuate nucleus-and in the paraventricular nucleus, but more in the former. It is suggested that somatosensory noxious input, or the partly noxious input evoked by electroacupuncture, activate the hypothalamo-pituitary-adrenocortical axis as in common forms of stress, but with a specific activation of the mediobasal hypothalamic nuclei and no stimulation of intermediate lobe cells. Opiate release from the pituitary gland may contribute to acupuncture analgesia or the intrinsic antinociceptive reactions triggered by noxious stimulation.

Adrenocorticotropic Hormone↗

PubMed1996

Attenuation of cyclic AMP production by carbamazepine.

The anticonvulsant carbamazepine is an effective treatment both for epilepsy and for bipolar affective disorder, but the molecular mechanism(s) underlying its therapeutic effects have not been identified. We have found that carbamazepine exerts significant inhibitory effects on the cyclic AMP (cAMP) generating system. Within the clinical therapeutic range (approximately 50 microM), carbamazepine inhibited both basal and forskolin-stimulated cAMP production, without having any significant effects on phosphodiesterase activity. Carbamazepine also exerted its inhibitory effects on the cAMP generating system in pertussis toxin-treated cells, suggesting that the action of carbamazepine was likely mediated through an inhibitory guanine nucleotide binding protein-independent mechanism. A forskolin affinity purification column was used to purify adenylyl cyclases from rat cerebral cortex, and we found that carbamazepine inhibited both basal and forskolin-stimulated activity of purified adenylyl cyclase. We also investigated the effects of carbamazepine on the levels of the transcription factor, cAMP response element binding protein in the phosphorylated (active) state, and found that carbamazepine significantly inhibited forskolin-induced phosphorylation of the cAMP response element binding protein. The data indicate that carbamazepine inhibits adenylyl cyclase activity as well as the downstream effects of activation of adenylyl cyclase.

1-Methyl-3-isobutylxanthine↗

PubMed1996

Autoregulation of iclR, the gene encoding the repressor of the glyoxylate bypass operon.

The aceBAK operon was partially induced by a multicopy plasmid which carried the promoter region of the gene which encodes its repressor, iclR. Gel shift and DNase I analyses demonstrated that IclR binds to its own promoter. Disruption of iclR increased the expression of an iclR::lacZ operon fusion. Although aceBAK and iclR are both regulated by IclR, aceBAK expression responds to the carbon source, while expression of iclR does not.

Bacterial Proteins↗

PubMed1996

The binding site of the IclR repressor protein overlaps the promoter of aceBAK.

In Escherichia coli, repression of the aceBAK operon is mediated by the IclR protein. We used an in vitro oligonucleotide selection technique to determine the consensus recognition sequence for MR. Mutational analysis confirmed the contribution of this sequence to repression in vivo and identified the -35 element of the promoter.

Bacterial Proteins↗

PubMed1996

Integration host factor amplifies the induction of the aceBAK operon of Escherichia coli by relieving IclR repression.

A binding site for integration host factor (IHF) was identified upstream of the aceBAK promoter. Under inducing conditions, IHF activates aceB::lacZ expression by opposing IclR repression. In contrast, IHF has little effect on aceB::lacZ expression under repressing conditions. The ability of IHF to relieve repression under inducing but not repressing conditions allows this protein to amplify the induction of aceBAK.

Acetates↗

PubMed1995

[Determination of diazepam concentration in maternal and fetal serum after intravenous administration during active phase of labor and its effects on neonates].

OBJECTIVE: To investigate the effects of diazepam on the neonates administered intravenously during the active phase in labour. METHODS: Sixty normal term parturients during the active phase of labour with the cervical dilatation to 3-5 cm were randomly divided into two groups, study group (n = 30) and control group (n = 30). In study group 10mg diazepam was administered intravenously. The concentrations of diazepam and demethyldiazepam in maternal and cord serum were measured by high performance liquid chromatography and neonatal arterial blood gas and Apgar scoring were determined immediately after birth. RESULTS: At delivery, the mean level of diazepam in umbilical cord serum was 947 +/- 314 micrograms/L, markedly higher than that of maternal serum which was 488 +/- 300 micrograms/L, while the mean levels of demethyldiazepam were not significantly different in maternal and fetal serum. There was no marked difference between the two groups in fetal arterial blood gas and acidbase status. However, the rates of respiratory depression and muscle tone inhibition in the neonates of study groups (63.3% and 26.7% respectively) were significantly higher than those in the control group (30% and 3.3% respectively) (P < 0.05, P < 0.01). CONCLUSIONS: Use of diazepam in pregnant women during active phase of labour may cause depression of neonatal respiration.

Adult↗