Biomedical subjects

L Cai

Publications and source records attributed to L Cai.

At least 127 records · Page 7Linked to original sources

PubMed1997

[Erythropoietin gene cloning and expression in S. cerevisiae].

Mature human erythropoietin gene was amplified from EPO cDNA by PCR methods. The PCR product was cloned into pUC18 plasmid at Sma I site, then precisely engineered into a intermidiate vector pSK43SB which were digested with Hind III, Mung bean nuclease, and Sal I. Then degest pSK43SB-EPO plasmid with EcoR I and Cla I, the EC fragment with an alpha-factor leading sequence, EPO gene and CYC1 terminater were produced. It was then cloned into a typical high efficiency episomal expression vector YEpHC8. Human EPO protein with highly mannose glycosylated was identified by Western blot methods in both secreted and in cells proteins. N-Glycosidase F digested secreted EPO can produce 20,000 EPO without N-glycosylation similar with that produced in cells.

Amino Acid Sequence↗

PubMed1997

[The simultaneous analysis of substance P, somatostatin, neurokinin and neurotensin by capillary zone electrophoresis].

Neuropeptides play an important part in several different areas of neurochemistry. Recently, capillary electrophoresis (CE) was developed for analysis of neuropeptides. This paper reports a capillary zone electrophoretic method for the simultaneous separation and quantitative determination of the substance P (SP), somatostatin (SS), neurokinin (NKA) and neurotensin (NT). The separation was performed on a 50microm x 60cm fused-silica capillary using 0.1mol/L at pH 2.7 phosphate as buffer. The eluted fractions were detected at 214nm.

Electrophoresis, Capillary↗

PubMed1996

Adaptive response to ionizing radiation-induced chromosome aberrations in rabbit lymphocytes: effect of pre-exposure to zinc, and copper salts.

Various stress conditions including exposure to low-dose radiation and low concentrations of chemical mutagens can induce an adaptive response to subsequent radiation-induced chromosome damage. In this study, the effect of pretreatment of rabbit lymphocytes with zinc or copper salts on radiation-induced chromosome damage was investigated. Pretreatment of rabbit peripheral lymphocytes with Zn (50 microM in vitro or 100 mumol/g body weight in vivo) resulted in resistance to gamma radiation (2.0 Gy)-induced chromosome aberrations such as dicentrics plus centric rings and cells with chromosome aberrations. On the other hand, pretreatment with Cu (50 microM in vitro) did not show any protective effect on radiation-induced chromosome damage in rabbit lymphocytes. However, the concentration of metallothionein increased in activated lymphocytes 24 h after in vitro pretreatment with both Zn and Cu. In addition, gamma-radiation-induced calf thymus DNA damage could be prevented directly by the addition of Zn-metallothionein in the cell-free system. These results suggest that the induction of zinc-metallothionein synthesis may act as one of the defensive mechanisms to the induction of cytogenetic adaptive response to ionizing radiation while copper-metallothionein did not show any radioprotective effect.

Adaptation, Physiological↗

PubMed1996

Compounds from Syzygium aromaticum possessing growth inhibitory activity against oral pathogens.

A crude MeOH extract of Syzygium aromaticum (clove) exhibited preferential growth-inhibitory activity against Gram-negative anaerobic periodontal oral pathogens, including Porphyromonas gingivalis and Prevotella intermedia. By means of bioassay-directed chromatographic fractionation, eight active compounds were isolated from this extract and were identified as 5,7-dihydroxy-2-methylchromone 8-C-beta-D-glucopyranoside, biflorin, kaempferol, rhamnocitrin, myricetin, gallic acid, ellagic acid, and oleanolic acid, based on spectroscopic evidence. The antibacterial activity of these pure compounds was determined against Streptococcus mutans, Actinomyces viscosus, P. gingivalis, and P. intermedia. The flavones, kaempferol and myricetin, demonstrated potent growth-inhibitory activity against the periodontal pathogens P. gingivalis and P. intermedia.

Anti-Bacterial Agents↗

PubMed1996

Androgen and pituitary control of penile nitric oxide synthase and erectile function in the rat.

Castration of adult male rats reduces by half the penile erectile response to electrical field stimulation (EFS) of the cavernosal nerve, and the activity of penile nitric oxide synthase (NOS). Both changes are prevented by androgen administration. We have now investigated whether other strategies of androgen ablation or competition may act as stronger inhibitors, and, if so, whether the stronger inhibition is due to the depletion of penile NOS content. Rats were castrated or left intact and were treated daily as follows: 1) intact, with the antiandrogen flutamide (25 mg/kg/day, i.p.); 2) castrated, with similar treatment; 3) castrated, with 17 beta-estradiol 3-benzoate (estradiol; via silastic tubing, s.c.). Additional groups of intact rats received injections of a GnRH antagonist (GnRHA, 1.25 mg/kg, s.c.), or were hypophysectomized and left untreated. Controls were untreated intact and castrated animals. After 7 days, rats were subjected to EFS, and the ratios between maximal intracavernosal pressure (MIP) and mean arterial pressure (MAP) were measured. Penile NOS activity and the contents of neuronal NOS (nNOS) and endothelial NOS (eNOS) were determined. Castration reduced the MIP:MAP ratio and penile NOS activity. Androgen receptor blockade with flutamide induced a similar response in intact rats. When flutamide treatment was combined with castration, the erectile response was nearly abolished, but NOS activity was not decreased below the values in castrated rats. Estradiol given to castrated rats and hypophysectomy or GnRHA treatment in intact rats diminished the erectile response below the level in castrated animals. In hypophysectomized rats, penile NOS activity fell below levels in castrated animals. contents of nNOS and eNOS were not significantly reduced by any treatment. These data suggest that penile erection in the rat is completely dependent on androgens, presumably because of their role in the maintenance of penile NOS activity and of other ancillary factors. However, only the complete blockade of residual androgen effects at the tissue level or a total androgen depletion can abolish the erectile response.

Androgen Antagonists↗

PubMed1996

[A clinical-pathological study on hypoxic ischemic encephalopathy in 124 cases of perinates].

OBJECTIVE: To explore the morphological feature, its pathogenesis and clinical significance of hypoxic-ischemic encephalopathy (HIE) through pathological examinations. METHODS: In addition to the routine pathologic examination, the brain tissues, including frontal lobus, temporal lobus, occipital lobus, hippocampus, capsula interna, cerebellum and medullary bulb, were sectioned and examined respectively. RESULTS: (1) 93.5% of cases had a history of direct or indirect asphyxia. (2) The main pathological alterations were edema, congestion, hyaline thrombosis, haemorrhage, degeneration and necrosis of nerve cells. Softening, calcification, cavity formation and fibrosis were also seen in some cases. CONCLUSIONS: The study suggested that congestion and edema of cerebral tissue were essential changes for the pathogenesis of HIE. Hemorrhage and necrosis were common pathological alterations. Fibrosis and cavitation were changes of convalescence. It is of primary importance to treat asphyxia during perinatal period for the prevention of HIE.

Asphyxia Neonatorum↗

PubMed1996

[A pathological survey of the therapeutic effect on experimental hypoxic-ischemic encephalopathy].

Rat models of the acute and recuperative phases of hypoxic ischemic encephalopathy (HIE) were established beginning by the 7th day after birth through ischemia and hypoxia. The prophylatic and therapeutic effects on experimental HIE were studied by the application of radix salviae miltiorrhizae, flunarizine and hyperbaric oxygen. Experimental data indicated that among these measures, radix salviae miltiorrhizae gave a better result and the pathological change in the prophylactic and therapeutic groups particularly the result of the latter one were light serious than those of the control group.

Animals↗

PubMed1996

[Prognosis of laryngeal carcinoma in youth].

The prognosis of 55 cases of laryngeal carcinoma in young people had been studied. The major factors mainly related to prognosis were smoking and misdiagnosis, while the tumour differentiation might be not so important. There was a high recurrence rate of 25.5% in the young patients and low 3- and 5-year survival rates of 47.4% and 34.5% respectively. The mortality of 1-year after surgery was 21.8%. The 3-and 5-year survival rates between total and partial laryngectomy made no difference. The survival rate of patients with positive node was lower. Early diagnosis is most important for young patients. Partial laryngectomy and neck dissection must be performed as far as possible.

Adult↗

PubMed1995

Construction and characterization of a bovine bacterial artificial chromosome library.

A bacterial artificial chromosome (BAC) library has been constructed for use in bovine genome mapping using constructed for use in bovine genome mapping using the pBeloBAC11 vector. Currently, the library consists of 23,040 clones, which achieves a 70% probability (P=0.70) of the library containing a specific unique DNA sequence. Sixty thousand clones, or about three haploid bovine genomes, will be required to achieve a 95% probability (P=0.95) of containing a unique sequence. An average insert size of 146 kb was estimated from the analysis of 77 randomly selected BAC clones produced by one or two rounds of size selection. The bovine DNA inserts proved to be very stable for at least 100 cell generations. No chimeric clones were detected among 11 large, size-selected BAC clones using fluorescence in situ hybridization (FISH) on metaphase bovine chromosomes. Thirty-three of 46 (72%) sequences were present in the library in at least one copy, which is consistent with the estimated 70% probability of this library containing a unique DNA sequence. A BAC clone as sequence-tagged sites for genetic mapping. These markers cosegregated, and no recombinants were detected in 193 informative meioses. Plasmid end rescue and the inverse polymerase chain reaction methods were used to rescue both ends of this BAC clone, and chromosome walking was performed using PCR primers designed within the end region sequences. Based on our experimental results, the BAC system provides a very useful tool for complex genome analysis.

3-Hydroxysteroid Dehydrogenases↗

PubMed1995

Metallothionein protects DNA from copper-induced but not iron-induced cleavage in vitro.

Iron and copper ions mediate generation of reactive oxygen radicals from O2 and H2O2 by the Fenton reaction: these radicals are capable of damaging DNA. We studied (a) the ability of these metals to induce double-strand breaks in DNA in vitro in the presence of H2O2 and ascorbic acid as donors of reactive oxygen, and (b) the ability of the metal-binding protein metallothionein (MT) to protect DNA from damage. Strand cleavage was measured by loss of fluorescence after binding to ethidium bromide and by increased mobility of DNA in agarose. The results show that Cu(II), Fe(II) and Fe(III) all can induce damage to calf thymus DNA under our experimental conditions. Cu(II)-induced DNA damage was dose-dependent and the degree of damage was proportional to the concentration of H2O2. On the other hand, DNA fragmentation was significant only in the presence of high concentrations of Fe(II) or Fe(III). Addition of Zn-MT to the reaction mixture prior to addition of Cu(II) inhibited fragmentation of DNA in a dose-dependent manner but had little effect on iron induced damage. Other proteins (histone or albumin) were not effective in protecting DNA from Cu-induced damage, as compared to Zn-MT. The formation of Cu(I) from Cu(II) in the presence of hydrogen peroxide and ascorbate was also inhibited by addition of Zn-MT. Thus, MT may protect DNA from damage by free radicals by sequestering copper and preventing its participation in redox reactions.

Animals↗

PubMed1995

A metal-dependent form of protein phosphatase 2A.

Highly purified bovine heart protein phosphatase 2A catalytic subunit lost virtually all of its activity during storage at -70 degrees. When the enzyme was preincubated with Co2+, over 35% of the original activity was restored. Freshly prepared protein phosphatase 2A purified from bovine heart was stimulated at least 3 to 4-fold by pretreatment with Co2+ or Mn2+. Activation by Co2+ appeared to be irreversible whereas activation by Mn2+ was partially reversed after the cation was chelated with excess EDTA/EGTA. The sensitivity of Co2(+)-stimulated protein phosphatase 2A to okadaic acid or inhibitor-2 was similar to that of spontaneously active protein phosphatase 2A. The enzyme was converted to a latent form by treatment with phosphate or pyrophosphate. The latent form was completely reactivated by preincubation with Co2+. These results demonstrate that protein phosphatase 2A, like phosphatase 1, can exist in a metal ion-dependent form and may represent a new mechanism for the regulation of protein phosphatase 2A activity.

Amino Acid Sequence↗

PubMed1995

Isolation and structural elucidation of pentacyclic triterpenoids from Maprounea africana.

Pentacyclic triterpenoids based on the taraxer-14-ene skeleton with a C-28 attached carboxylic acid group have been isolated from the roots of Maprounea africana. These compounds were identified as 1 beta,2 alpha-dihydroxyaleuritolic acid 2,3-bis-p-hydroxybenzoate [1], 2 alpha-hydroxyaleuritolic acid 3-p-hydroxybenzoate [2], 2 alpha-hydroxyaleuritolic acid 2,3-bis-p-hydroxybenzoate [4], aleuritolic acid 3-p-hydroxybenzoate [5], aleuritolic acid [6], and aleuritolic acid 3-acetate [7]. Compounds 1 and 2 are new triterpene esters. Compound 3 was previously reported as 7 beta-hydroxymaprounic acid 3-p-hydroxybenzoate [13]. However, based on detailed nmr studies, its structure has been revised.

Magnetic Resonance Spectroscopy↗

PubMed1995

Mechanistic evaluation of new plant-derived compounds that inhibit HIV-1 reverse transcriptase.

Swertifrancheside [1], a new flavonone-xanthone glucoside isolated from Swertia franchetiana, 1 beta-hydroxyaleuritolic acid 3-p-hydroxybenzoate [2], a triterpene isolated from the roots of Maprounea africana, and protolichesterinic acid [3], an aliphatic alpha-methylene-gamma-lactone isolated from the lichen Cetraria islandica, were found to be potent inhibitors of the DNA polymerase activity of human immunodeficiency virus-1 reverse transcriptase (HIV-1 RT), with 50% inhibitory doses (IC50 values) of 43, 3.7, and 24 microM, respectively. They were not cytotoxic with cultured mammalian cells. The kinetic mechanisms by which compounds 1-3 inhibited HIV-1 RT were studied as was their potential to inhibit other nucleic acid polymerases. Swertifrancheside [1] bound to DNA and was shown to be a competitive inhibitor with respect to template-primer, but a mixed-type competitive inhibitor with respect to TTP. On the other hand, 1 beta-hydroxyaleuritolic acid 3-p-hydroxybenzoate [2] and protolichesterinic acid [3] were mixed-type competitive inhibitors with respect to template-primer and noncompetitive inhibitors with respect to TTP. Therefore, the mechanism of action of 1 beta-hydroxyaleuritolic acid 3-p-hydroxybenzoate [2] and protolichesterinic acid [3] as HIV-1 RT inhibitors involves nonspecific binding to the enzyme at nonsubstrate binding sites, whereas swertifrancheside [1] inhibits enzyme activity by binding to the template-primer.

4-Butyrolactone↗

PubMed1995

Induction of a cytogenetic adaptive response in germ cells of irradiated mice with very low-dose rate of chronic gamma-irradiation and its biological influence on radiation-induced DNA or chromosomal damage and cell killing in their male offspring.

In earlier studies we have shown that either a single exposure or multiple exposures to a low dose of X-rays (0.05 Gy) induced a significant cytogenetic adaptive response in mouse germ cells. In this paper, a very low-dose rate (20 microGy/min) of chronic 60Co gamma-irradiation was used to pre-irradiate mice for 40 days. Then, another 40 days later, these mice were treated with a subsequent large dose of X-irradiation, followed 24 h later by cytogenetic analysis of their spermatocytes. Analysis for radiation-induced DNA and chromosomal damage was also carried out in splenocytes, bone marrow cells and spermatocytes of the offspring of mice adapted by the low-dose rate of chronic gamma-irradiation. Results demonstrated that (i) cumulative gamma-irradiation (1.10 Gy) at the dose rate 20 microGy/min induced a marked cytogenetic adaptive response in the mouse germ cells (stem spermatogonia); (ii) the sensitivity of offspring's bone marrow cells and spermatocytes to 1.5 Gy X-ray-induced chromosome aberrations was not influenced by the low-dose radiation delivered to paternal germ cells; (iii) either constitutive or post-irradiation DNA repair capacity (UV-induced unscheduled DNA synthesis, UDS) was not modified in the offspring's splenocytes; (iv) the sensitivity of the offspring's splenocytes to radiation-induced cell killing was also not altered. These results suggest that low-dose radiation delivered to the male parents with a significant induction of cytogenetic adaptive response in their germ cell does not likely cause any risk of damaging effects to the offspring of those irradiated male mice.

Adaptation, Physiological↗

PubMed1995

Restoration of normal adult penile erectile response in aged rats by long-term treatment with androgens.

In order to determine whether the aging-related impairment of penile erection can be corrected by exogenous androgens, 20-mo-old ("old") rats received implants of Silastic tubing containing either testosterone (T) or dihydrotestosterone (DHT). Untreated aged-matched rats were used as reference controls, and values were compared with those obtained previously for 5-mo old ("adult") rats. After 45 days, groups of six rats were submitted to electrical field stimulation of the cavernosal nerve (EFS), and the intracavernosal pressure and mean arterial pressure (MAP) were recorded. Compared to the untreated old rats, the old animals receiving either T or DHT showed a similar and significant (p < 0.01) increase (56%) in the maximal intracavernosal pressure (MIP) to a level slightly above that of the untreated adult rats. The response was sensitive to the nitric oxide synthase (NOS) inhibitor Nw-nitro-L-arginine methyl ester (L-NAME). The MIP:MAP ratio in the treated animals was nearly double that found in the old untreated controls. Penile NOS activity was measured by the [3H]L-arginine-citrulline conversion, and neuronal NOS (nNOS) levels were estimated by a semiquantitative Western blot assay with antibodies against human nNOS. No significant variations were observed in either NOS levels or NOS activity between the treated and untreated old rats. These results suggest that aging-related erectile dysfunction in the intact rat may be responsive to androgen therapy and that this correction is not associated with an increase in the basal levels of penile NOS, in contrast to what occurs in castrated rats.

Aging↗

PubMed1995

Reduction of penile nitric oxide synthase in diabetic BB/WORdp (type I) and BBZ/WORdp (type II) rats with erectile dysfunction.

Erectile dysfunction occurs frequently in human diabetes, and it is sometimes associated with hypogonadism. These conditions also develop in a model of insulin-dependent (type I) diabetes, the BB/WORdp (diabetic prone) rat but have not yet been investigated in the model of insulin-resistant (type II) diabetes, the BBZ/WOR rat. It is also unknown whether diabetes-related impotence is due to reduced levels of the mediator of penile erection, nitric oxide, caused by a decrease of nitric oxide synthase (NOS) in the penis. To clarify these questions, groups (n = 5-6) of diabetic BB/WORdp (insulin-maintained) and BBZ/WOR rats were age-matched with diabetic-resistant BB/WORdr and non-diabetic BB/WORdp rats and submitted to determinations of serum glucose, testosterone, and penile reflexes (cups and flips). Erectile dysfunction was found in all of type I and in most of type II diabetic animals (glycemias of 25.0 and 31.1 mM), at the selected mean ages of 310 and 180 days old, respectively. This was evidenced by over 95% decreases of erectile reflexes in both types of diabetes and was accompanied by 75% reduction of serum testosterone. Soluble NOS activity was measured in penile tissue from the diabetic rats with impaired erectile reflexes and in the corresponding controls, by the (3H)-L-arginine/citrulline conversion assay. The neuronal NOS isoform (nNOS) content was determined by a semiquantitative western blot assay. Both types of diabetes showed a marked decrease of penile NOS activity (74 and 55%, respectively), and a lower reduction of penile nNOS content (47 and 33%, respectively). No endogenous NOS inhibitor was detected in the diabetic type I penile cytosol by cross-mixing NOS activity assays. Our data support a common etiology for erectile dysfunction present in rats with types I and II diabetes mellitus and suggest that the etiology is related to a decrease of penile NOS derived in part from serum androgen deficiency.

Animals↗

PubMed1995

Mild hyperthermia can induce adaptation to cytogenetic damage caused by subsequent X irradiation.

Many low-level environmental agents are able to induce an increased resistance to subsequent mutagenic effects induced by ionizing radiation. In this paper, an induced cytogenetic adaptation to radiation in human lymphocytes was studied with mild hyperthermia as the adaptive treatment and compared with that induced by low-dose radiation. We found that this adaptation could be induced not only in PHA-stimulated human lymphocytes (at 14, 38 and 42 h after addition of PHA), but also in unstimulated G0-phase cells (before addition of PHA) by mild hyperthermia (41 degrees C for 1 h) as well as 50 mGy X rays. When the two adaptive treatments were combined, no additive effects on the magnitude of the adaptation induced were observed, suggesting that low-dose radiation and hyperthermia may share one mechanism of induction of adaptation to cytogenetic damage. Some mechanisms which may be involved in the induction of adaptation to cytogenetic damage by low-dose radiation are discussed and compared with the effects of mild hyperthermia in inducing thermotolerance and radioresistance.

Adaptation, Physiological↗

PubMed1995

[Influence of radiotherapy and chemotherapy on the function of malignant tumor patients and regulation function of acupuncture].

The observation on the indexes of cortisol, estradiol, estriol and testosterone showed that incretory function of malignant tumor patients had different extent of pathologic changes, after radiotherapy and chemotherapy, making the change strengthened. Acupuncture can regularize this disorder of incretory function of patients treated with radiotherapy and chemotherapy to some extent.

Acupuncture Therapy↗