Clustering of gout-related comorbidities and their relationship with gout flares: a data-driven cluster analysis of eight comorbidities.

OBJECTIVES: To study the aggregation of multiple comorbidities in people with gout and explore differences in prognosis of gout flares among different subgroups. METHODS: Hierarchical clustering was performed to identify homogeneous subgroups among 2639 people with gout using eight comorbidities. A one-year follow-up of acute gout flares in 463 of these people was conducted; the incidence and the timing of gout flares in each cluster were assessed to explore prognosis of gout flares. Binary logistic regression was applied to assess factors associated with gout flares. RESULTS: In baseline study, we identified five subgroups (C1-C5). C1 (n = 671, 25%) was characterized by isolated gout with few comorbidities. C2 (n = 258, 10%) were all obese. Almost all people in C3 (n = 335, 13%) had diabetes (99.7%). All people in C4 (n = 938, 36%) had dyslipidemia. C5 (n = 437, 17%) had the highest proportion of cardiovascular disease (CVD, 53%), chronic kidney disease (CKD, 56%), and cancer (7%). In follow-up study, C5 had the highest incidence (71.9%) and earliest onset (median 3 months) of gout flares. C2 had the lowest incidence (52.1%) and the latest onset (median 10 months) of gout flares. The highest relative risk for gout recurrent was seen for C5 (OR = 2.09). Other factors associated with the risk of gout flares were age at diagnosis of gout, duration of gout, presence of tophi, and smoking ≥ 20 cigarettes/day. CONCLUSIONS: We clustered people with gout into five groups with varying comorbidities. People with CVD, CKD, and cancer had the highest risk of gout flares and should receive comprehensive care.

The effect of the major components of Salvia Miltiorrhiza Bunge on bone marrow cells.

Salvia Miltiorrhiza Bunge (SMB), a traditional Chinese medicinal herb, has been alleged to support bone healing. However, the effects of the isolated major components of SMB on osseous cells and their corresponding effective doses are still unclear. In the present study, the effects of three components of SMB, including tanshinone IIA (Ts), salvianolic acid B (salB) and protocatechuic aldehyde (Pca), on mesenchymal bone marrow cells with the potential for osteoblastic differentiation were investigated. Various concentrations of Ts, salB and Pca were added to a rat bone marrow cell culture. The total metabolic activity and differentiation of bone marrow cells were evaluated by a metabolic assay and alkaline phosphatase (ALP) activity, respectively. The morphology and number of cells was observed by phase contrast microscopy and fluorescent microscopy after propidium iodide staining, respectively. Ts suppressed the growth and differentiation of bone precursor cells. SalB exhibited a biphasic effect: the high concentration of 160 microg/mL significantly depressed the population of bone marrow cells, however, lower concentrations (3-80 microg/mL) enhanced the total metabolic activity and their ALP expression. Pca suppressed the bone marrow cell population in a dose-dependent manner. Therefore, SalB has the potential to ameliorate bone healing by stimulating both the total metabolic activity and ALP activity of osteoblastic cells. Aqueous extracts, which preferably contain salB over Pca and are free of Ts therefore are recommended for bone formation.

Radiation-mediated control of drug delivery.

Clinical trials of radiotherapy to control drug delivery were initiated in 1999 at Vanderbilt University. The initial studies exploited the findings that platelets are activated in tumor blood vessels after high-dose irradiation as used in radiosurgery and high-dose-rate brachytherapy. Platelets labeled with 111In showed binding in tumor blood vessels. However, the platelet labeling process caused platelets to also accumulate in the spleen. That clinical trial was closed, and subsequent clinical trials targeted protein activation in irradiated tumor blood vessels. Preclinical studies showed that peptide libraries that bind within irradiated tumor blood vessels contained the peptide sequence Arg-Gln-Asp (RGD). RGD binds to integrin receptors (e.g., receptors for fibrinogen, fibronectin, and vitronectin). We found that the fibrinogen receptor (GPIIb/IIIa, alpha2bbeta3) is activated within irradiated tumor blood vessels. RGD peptidemimetics currently in clinical trials include GPIIb/IIIa antagonists and the platelet-imaging agent biapcitide. Biapcitide is an RGD mimetic that is labeled with 99Tc to allow gamma camera imaging of the biodistribution of the GPIIb/IIIa receptor in neoplasms of patients treated with radiosurgery. This study has shown that the schedule of administration of the RGD mimetic is crucial. The peptide mimetic must be administered immediately before irradiation, whereas the natural ligands to the receptor compete for biapcitide binding if biapcitide is administered after irradiation. The authors currently are conducting a dose deescalation study to determine the threshold dosage required for RGD mimetic binding to radiation activated receptor. Radiation-guided clinical trials have been initiated by use of high-dose-rate brachytherapy. In a separate trial, the pharmacokinetics of radiation-inducible gene therapy are being investigated. In this trial, the radiation-activated promoter Egr-1 regulates expression of the tumor necrosis factor alpha gene, which is administered by use of the attenuated adenovirus vector. The Ad.Egr-TNF (ADGV) gene is administered by intratumoral injection of vector followed by irradiation in patients with soft-tissue sarcomas. This review highlights recent findings in these phase I pharmacokinetic studies of radiation-controlled drug delivery systems.

Agouti-related protein is a mediator of diabetic hyperphagia.

To explore the role of agouti-related protein (AGRP) in diabetic hyperphagia changes in hypothalamic AGRP mRNA levels were examined in diabetic rats. Rats rendered diabetic by streptozotocin displayed marked hyperglycemia (blood glucose 456.0+/-8.4 mg/dl versus 71.8+/-1.9 mg/dl) and hyperphagia (36.9+/-1.0 g/day versus 22.0+/-0.4 g/day), that was associated with a 286.6+/-4.4% increase in hypothalamic AGRP mRNA and a 178.9+/-13.5% increase in hypothalamic NPY mRNA. Insulin treatment of diabetic rats partially corrected blood glucose (147.4+/-13.1 mg/dl) and ameliorated hyperphagia (26.6+/-2.0 g/day). Insulin replacement was also associated with a return of hypothalamic AGRP mRNA (111.7+/-8.3% of controls) and NPY mRNA (125.0+/-8.9% of controls) from the elevated levels that were observed in untreated diabetic rats. In contrast to insulin treated rats, sodium orthovanadate treated diabetic rats remained significantly hyperglycemic (361.5+/-12.5 mg/dl). However, despite their persistent hyperglycemia, orthovanadate treated diabetic rats were still observed to have a significant reduction of hypothalamic AGRP mRNA (138.7+/-11.4%) and NPY mRNA (129.9+/-9.8%). Simultaneous measurement of serum leptin revealed suppressed levels in both untreated diabetic (0.5+/-0.1 ng/ml) and sodium orthovanadate treated rats (0.5+/-0.1 ng/ml) compared to non-diabetic controls (2.1+/-0.1 ng/ml). These data indicate that AGRP is a mediator of diabetic hyperhpagia and suggest that insulin can directly influence hypothalamic AGRP and NPY mRNA expression.

[Protective effect of Panax quinquefolium 20s-proto-panaxdiolsaponins on acute myocardial infarction in dogs].

OBJECTIVE: To study the protective effects of Panax quinquefolium 20s-protopanaxdiolsaponins extracted from leaves of P. quinquefolium (PQDS) on acute myocardial infarction(AMI) in dogs. METHOD: The parameters of myocardial infart size, the serum CK and LDH activity, myocardial metabolism, free radicals and coronary circulation etc were determined by using the model of ligation of LAD in the anaesthetized open-chest dogs. RESULT: In dogs treated with PQDS(in a dosage of 10 and 20 mg.kg-1 i.v. infusion), the myocardial infarct size, the activity of serum CK, LDH and the contents of serum FFA and LPO were decreased, whereas the activity of serum SOD and GSH-Px increased markedly. At the same time, myocardial blood flow was increased and coronary vascular resistance decreased significantly. CONCLUSION: PQDS has protective effect on myocardial ischemia by modifying metabolic dysfunction of FFA, inhibiting oxygen free radical mediated peroxidation of membrane lipids, enhancing endogenous antioxidase activity and increasing myocardial blood supply.

Agouti-related protein-like immunoreactivity: characterization of release from hypothalamic tissue and presence in serum.

A novel RIA was used to examine the release of agouti-related protein-like immunoreactivity (AGRP-LI) from perfused rat hypothalamic tissue slices and to characterize AGRP-LI in rat serum. A continuous low level basal AGRP-LI release was observed from hypothalami of rats fed ad libitum before the rats were killed. Basal AGRP-LI release was 3-fold greater in rats fasted 48 h. In fasted animals leptin dose-dependently suppressed basal AGRP-LI release. In fed animals no change in basal AGRP-LI release was detected in response to 10(-6) M alpha-MSH, orexin B, melanin-concentrating hormone, or serotonin. HPLC analysis of AGRP-LI in rat serum identified a single peak that eluted in close proximity to synthetic AGRP (87-132) and mouse [Leu127Pro]AGRP and that was identical to the peak seen in hypothalamic and adrenal tissue extracts. The serum concentration of AGRP-LI in rats fed ad libitum was 0.865+/-0.323 nmol/liter (mean +/- SE). Food deprivation resulted in a slow, but statistically significant rise in serum immunoreactivity at 48 h [1.174+/-0.118 nmol/liter (mean +/- SE)]. Bilateral adrenalectomy did not change serum levels of AGRP-LI. These studies demonstrate that in the rat there are different levels of basal hypothalamic AGRP-LI release in fed and fasted states and that in the fasted rat this release can be profoundly suppressed by leptin. These studies also suggest that AGRP is present in the systemic circulation of rats.

[Effect of motherwort herb on the myoelectric activity of uterus in rats].

OBJECTIVE: To investigate the effect of motherwort herb on the myoelectric activity of uterus in rats. METHOD: A pair of bipolar Ag-AgCl electrode was implanted on the serosal surface of the uterus in rats, motherwort herb was injected intra-abdominal cavity in rats in dose of 0.1 ml, 0.2 ml and 0.4 ml respectively, and the changes of uterine myoelectric activity were observed. RESULT: Motherwort herb significantly increased the frequency and average amplitude of uterus slow waves of in rats. The frequency and maximum amplitude of single waves also obviously increased. The time course of outbreak waves was prolonged, the series intervals were reduced and the maximum amplitude in each outbreak was increased. CONCLUSION: The exciting effect of motherwort herb on the uterine smooth muscle may have something to do with the alteration of the ion concentration in relation to myoelectric activity, resulting in the increase of myoelectric activity of pacesetter cells as well as in the acceleration of depolarization of spike activity.

[Effect of emodin on human kidney fibroblast proliferation].

OBJECTIVE: To study the effect of emodin on proliferation of human kidney fibroblasts in vitro. METHODS: After human kidney fibroblasts were cultured, isolated and identified, both the effects of five different concentrations of emodin (10, 30, 50, 80 and 100 micrograms/ml) on 3H-TdR incorporation, and the effects of three different concentrations of emodin (10, 50 and 100 micrograms/ml) on cell cycle by flow cytometry were investigated. RESULTS: The exposure of human kidney fibroblasts to emodin (10-100 micrograms/ml) caused a dose-dependent reduction in 3H-TdR (r = 0.995, P < 0.01), and could delay the progress of human kidney fibroblasts from G1 to S phase. CONCLUSIONS: Emodin inhibited the proliferation of human kidney's fibroblasts by inhibiting the cell DNA synthase and delaying the progress of cell cycle. These findings might provide part of experimental basis for the clinical use of emodin.

[Effect of yuanhua on isolated uterine strips in unpregnant rats].

OBJECTIVE: To explore the mechanism on the effect of Yuanhua on the contractile activity of isolated uterine strips in rats under artificial oestrum. METHOD: Isolated strips were suspended in a tissue chamber in which Yuanhua was added with some antagonists of inhibitors or without. RESULT: Yuanhua could excite the uterine smooth muscle strips in a dose-dependent manner, but atropine, phentolamine, hexamethonium and diphenhydramine could not inhibit this exciting action. This exciting effect could be inhibited completely by verapamil and decreased by indomethacin(inhibitor of prostaglandin synthase). CONCLUSION: The exciting effect of Yuanhua on un-pregnant rats in vitro might be related to an action on the cell membrane Ca2+ channel as well as to a partial stimulation of synthesis and release of PG.

Microcalorimetric study of the toxic effect of selenium on the mitochondrial metabolism of Cyprinus carpio liver.

The metabolic thermograms and heat output of mitochondria isolated from carp liver have been determined by using an LKB bioactivity monitor. The thermogram can be divided into four parts: the lag phase, active recovery phase, stationary phase, and decline phase. The thermokinetic equation was established for the active recovery and decline phase of metabolism as follows: dP/dt = k(m)P (1-SP). The rate constants k1 and k2 of two phases of active recovery and decline phase have been also calculated. The metabolism activity of mitochondrial inhibited by a high concentration of trace element selenium has been studied. The metabolic heat released, time of each phase, and rate constants can be significantly influenced by excess of selenite added. These results suggested that a high concentration of selenium can damage the structure and function of mitochondria, and thus influence their metabolism.

[Studies on function of vegetative nervous system in cardiopathic patients with heart-qi deficiency syndromes and heart-yin deficiency syndromes].

To explore the changes of the function of vegetative nervous system in the patients with the syndromes of Heart-Qi Deficiency (HQD) and Heart-Yin Deficiency (HYD), 53 cases of heart diseases were divided into HQD and HYD, the duration between heart beats, differences of blood pressure in the lying and standing position, and urine-catecholamine/24 hrs was determined separately, and compared with control group. The results showed that the duration between heart beats and heart rate in the lying position in those patients of HQD didn't manifest significant difference with control group. But in 78.9% of patients, the differences of heart rate in expiration/inspiration, surpassed 15 beats per minute, and the proportion of 30/15 was less than 1.03. Compared with control group, the difference was significant (P < 0.01). Meantime, the differences of heart rate of lying/standing position was less than 15 beats per minute, and the difference of blood pressure had an increasing tend in 47.4% of the cases. Those patients of HYD didn't show any difference in the above indexes. Urine-catecholamine/24 hrs in both of the two groups surpassed that of control group (P < 0.01). In short, this study showed that syndromes of HQD and HYD both displayed disturbed function of the vegetative nervous system, and their types and severities were correlated with the difference of Syndromes.

Heat shock effects on phosphorylation of protein synthesis initiation factor proteins eIF-4E and eIF-2 alpha in Drosophila.

Heat shock of mammalian cells causes changes in initiation factor phosphorylation that likely contribute to or cause the translation reprogramming characteristic of heat shock. In these investigations we have carried out a parallel analysis of Drosophila, focusing on eIF-4E and eIF-2 alpha. eIF-4E plus associated proteins was purified from lysates by m7GTP-Sepharose chromatography. A minor fraction (< 10%) of eIF-4E is phosphorylated under normal growth conditions, and phosphorylation decreases during heat shock. Drosophila eIF-2 alpha has been identified by in vitro translation of T7 RNA polymerase-transcribed mRNA, and immunoblotting with anti-Drosophila eIF-2 alpha antiserum. 32P-labeling analysis (unfractionated cell lysates and immunoprecipitates) detects phosphorylated eIF-2 alpha, whose amount increases approximately 2-3-fold upon heat shock. Immunoblotting analysis of two-dimensional gel-resolved proteins to determine the mass fraction of eIF-2 alpha phosphorylated detects a single eIF-2 alpha spot in both normal temperature and heat shocked cells, indicating less than 5% phosphorylation after and before heat shock. Staining quantification is consistent with this low prevalence. A major phosphoprotein which copurifies with eIF-4E on m7GTP-Sepharose shows decreased overall phosphorylation and decreased association with eIF-4E following heat shock. Several distinctive characteristics of this phosphoprotein suggest it is Drosophila eIF-4B.

Effects of site-directed mutagenesis on the serine residues of human lecithin:cholesterol acyltransferase.

Lecithin:cholesterol acyltransferase (LCAT) is a serine protease-type enzyme that esterifies cholesterol in human plasma and is activated by apolipoprotein A-I in high-density lipoproteins. LCAT contains 22 serine residues, including Ser181, which is thought to be part of the catalytic site. In order to determine the importance of these serine residues in LCAT, we prepared six LCAT mutants: LCAT (Ser19-->Ala), LCAT (Ser181-->Gly), LCAT (Ser208-->Ala), LCAT (SEr216-->Ala), LCAT (Ser225-->Ala) and LCAT (Ser383-->Ala). We also replaced the adjacent asparagine residues in two additional mutants, LCAT (Ser19-->Ala, Asn20-->Thr) and LCAT (Ser383-->Ala, Asn384-->Thr), in order to ascertain the effect of the serines on N-glycosylation. The mutant complementary DNA (cDNA) were subcloned into a eukaryotic expression vector (pSG5) and expressed in COS-6 cells. By polymerase chain reaction analysis, LCAT-specific messenger RNA (mRNA) was found in all mutant and wild-type transfectants. Western blot analysis revealed LCAT-specific bands in media and lysates of the transfected cells. With two exceptions, the amounts of LCAT mass secreted by the transfectants were similar to that of the wild type (mean, 90% mass of wild type; range, 34-138%). Except for LCAT (Ser181-->Gly), which was inactive, the specific activities of the remainder of the mutant enzymes were also similar (mean 95% activity of wild type; range, 65-169%). These results indicate that Ser181 is part of the catalytic site and that stereoconservative substitutions for serines have minor effects on the synthesis, secretion and specific activities of human LCAT.

[Experimental study on anti-senility effects of centipede].

Our experiment has shown that the aqueous extract of Centipede can markedly decrease the contents of lipid peroxide in the serum as well as lipofuscin in the liver tissues and brain tissues of rats, enhances the activity of superoxide dismutase in erythrocyte and glutathione in blood, but has no action on the lipofuscin in the heart. In addition, the extract is able to increase the weight of spleen and thymus.

[Effect of Equisetum hyemale on experimental hyperlipemia in rats and its toxic test].

The results of an experimental study in rats fed with Equisetum hyemale and hyperlipid food have proved that inhibiting effects on the elevation of triglyceride and cholesterol can be obviously observed in both high and low doses of Equisetum. The study also shows that Equisetum hyemale can antagonize the hyperlipemia in rats. The acute toxic test has proved its low toxicity.

[Pharmacological studies on the volatile oil isolated from the leaves of Pinus pumila (Pall.) Regel].

The sedative, analgesic, antipyretic and anti-inflammatory activities of the volatile oil isolated from the leaves of Pinus pumila are reported, the volatile oil being administered ip or po. The ip LD50 in mice was 0.577 +/- 0.056 ml/kg.

[The relation of typing of gastric carcinoma according to traditional Chinese medicine theories and clinical pathologic classification].

102 cases of gastric carcinoma which have been removed by partial gastrectomy and have pathological diagnoses and follow-up results are reported. The relationship between their preoperational syndromes of TCM and pathological, TNM classification and post-operational survival rate has been studied. There is indication that the type of Ganwei Buhe (the dispersing function of the liver is disturbed and affects the stomach) manifests the earlier stage of the gastric carcinoma and the majority of cases are in stage 1 or 2 in TNM, the 5-year survival rate with surgery being 46%. The majority of cases of the types of Pishen Yangxu (deficiency of energy in the Spleen and Kidney), Yinxu Neire (the febrile syndrome of the viscera caused by the insufficient primordial energy) and Qixue Shuangkui (deficiency of both energy and blood) are in stage 3 or 4 in TNM, the 5-year survival rate with surgery differently being 5,8,12% respectively. The type of Qizhi Xueyu (energy stagnancy and blood stasis) is between the abovementioned two groups, with a 5-year survival rate of 40%.