CAMK4 gene variation is associated with hypertension in a Uygur population.

Considering that calcium/calmodulin-dependent kinase 4 (CAMK4) plays a pivotal role in blood pressure regulation, we investigated the association between a CAMK4 polymorphism (rs10491334) and hypertension in the Han, Kazak, and Uygur ethnic groups. We studied 1224 patients with hypertension and 967 normotensive controls classified into three ethnic groups (Han, Kazak, and Uygur). The rs10491334 polymorphism was genotyped using a TaqMan® 5'-nuclease assay. In the Uygur group, the T-allele frequency in patients with hypertension was twice that of the controls (12.5 vs 6.38%), and T-allele carriers had a significantly increased risk of hypertension compared with non-carriers (odds ratio = 2.200; 95% confidence interval = 1.473-3.285, P < 0.001). However, no significant correlation was found in the Han and Kazak groups. The T-allele of rs10491334 in CAMK4 was associated with hypertension in the Uygur group.

Activity of corilagin on post-parasiticide liver fibrosis in Schistosomiasis animal model.

This study investigates the effects and possible molecular mechanisms of corilagin extraction on prevention of Schistosoma japonicum ova-induced granulomas and liver fibrosis. As a result, under a light microscope, when compared to a model group, the corilagin group showed smaller granulomas, less liver cell denaturation and less inflammatory cell infiltration, and the connective tissues were significantly decreased. By Masson staining, the liver sections from the corilagin group showed less collagen distributed around granulomas, decreased liver fibrosis in the portal tracts and less formed interlobular tissue. The expression of hydroxyproline, IL-13 in liver and GATA3 in spleen in the model group was significantly higher than that in the normal group (P less than 0.05 or 0.01), while the level of hydroxyproline, IL-13 and GATA3 in the corilagin group were significantly lower than that in the model group (P less than 0.05). In conclusion, corilagin extraction can decrease the level of Th2-associated profibrotic cytokine IL-13, and down-regulate the transcription of GATA3 mRNA in spleen cells, which alleviate the hepatic fibrosis caused by egg granuloma in Schistosoma japonicum infection.

Pharmacokinetics and metabolism of 1,5-dicaffeoylquinic acid in rats following a single intravenous administration.

1,5-Dicaffeoylquinic acid (1,5-DCQA) is a potentially important HIV-1 integrase inhibitor widely distributed in many plants. To characterize the pharmacokinetic and metabolic properties of 1,5-DCQA in rats following single intravenous administration (160 mg/kg), the plasma concentrations of 1,5-DCQA were measured by high-performance liquid chromatography (HPLC) and the metabolites formed in urine were identified by liquid chromatography-mass spectrometry (LC-MS) in parallel to diode-array detection (DAD). The results showed that the concentrations of 1,5-DCQA in plasma declined rapidly in a biphasic manner with a mean terminal half-life (t(1/2)) of 1.40 h. The mean clearance (CL) and the apparent volume of distribution (Vd(B)) of 1,5-DCQA were 0.44l/h/kg and 0.89l/kg, respectively. A total of 15 metabolites in rat urine were identified, including four isomeric O-mono-methylated (M1-M4), six isomeric O-di-methylated (M5-M10), one isomeric O-mono-methyl-glucuronidated (M11) and four isomeric O-di-methyl-glucuronidated (M12-M15) metabolites. The O-methylation positions of three important metabolites (M1, M2 and M5) were determined (3''-, 3'-, and 3',3''-) by comparing with synthesized standards. These results suggested that the disappearance of 1,5-DCQA from plasma was rapid, and that its quick urinary excretion and extensive metabolism, including methylation and glucuronidation, were two factors causing its rapid elimination from the circulation.

Pharmacokinetics of (+/-)-, (+)-, and (-)-gossypol in humans and dogs.

Pharmacokinetic parameters of (+/-)-, (+)-, and (-)-gossypol were determined in humans and dogs after a single oral or intravenous dose. Mean (+/- SD) oral bioavailability of (+/-)-gossypol in dogs was 30.9% +/- 16.2%. Studies in dogs who received single intravenous injections revealed that the elimination t1/2 and volume of distribution of (+)-gossypol were five and six times those of (-)-gossypol, respectively, whereas total body clearance and the AUC of the two enantiomers were similar. Data from men receiving the compounds orally show that the average peak plasma concentration and the AUC of (+)-gossypol are significantly greater than those of the (-)-isomer. The rate constants of alpha, beta, ka, k21, and k10 for (-)-gossypol are significantly greater than those for (+)-gossypol, indicating higher rates of mass transfer of the (-)-species. In humans the elimination t1/2 of (+)-gossypol was 29 times as that of (-)-gossypol, a difference that is more striking than that found in dogs. The elimination t1/2 of (+/-)-gossypol in humans averages 286 +/- 179 hours.

Beneficial effects of N-acetylcysteine and cysteine in stunned myocardium in perfused rat heart.

1. The objective of this study was to evaluate the effects of three sulphydryl (SH) compounds, N-acetylcysteine (NAC), cysteine (Cys) and cystamine, on functional recovery and ventricular arrhythmias (VF) in stunned myocardium in the isolated perfused heart of the rat. 2. Hearts (n = 7-8 per group) were perfused by the Langendorff procedure for 20 min to stabilize and then assigned to one of five groups: saline, sham, NAC, Cys and cystamine. After the stabilizing period, the drugs (at 3.6 microM min-1) or their vehicle (saline) were infused into coronary vessels throughout the experimental period. Ten min after administration of drugs, the left anterior descending coronary artery (LAD) was ligatured for 20 min and then untied to reperfuse for 30 min. In the sham group, a ligature was placed around the LAD but not tied. 3. NAC and Cys had a significant effect in attenuating myocardial stunning: the percentage recovery of rate-pressure product measured 30 min after reperfusion as an index of heart function, was improved with the NAC (98.3 +/- 4.5) and Cys groups (104.0 +/- 6.5) compared with the saline (only 73.6 +/- 3.8, P < 0.01) group. Cystamine did not show these beneficial effects. This may be due to the difference in chemical structure between NAC, Cys and cystamine since the latter does not have a free SH group with a disulphide bond formed. This phenomenon suggests that a free SH group is essential for the protective effects of compounds like NAC and Cys in myocardial injury. 4. NAC and Cys prevented the fall in coronary flow during the LAD occlusion and enhanced coronary flow during reperfusion but cystamine did not have such a beneficial effect. 5. The incidence of VF in the saline, cystamine, Cys and NAC groups was 6/8 (75.0%), 4/7 (57.1%), 3/8 (37.5%) and 2/7 (28.6%), respectively, and no significant differences (P > 0.05) were noted between the saline- and drug-treated groups. 6. An in vitro study with electron spin resonance indicated that Cys effectively scavenged the hydroxyl radical (-OH) generated by Fenton's reaction but did not scavenge superoxide generated in an irradiated riboflavin system. NAC and cystamine showed a scavenging effect on -OH to a certain extent but this effect did not reach statistical significance (P > 0.05 vs saline). 7. Our results demonstrate that NAC and Cys treatment before ischaemia and reperfusion can reduce myocardial stunning. This beneficial effect may be mainly due to their ability to preserve and enhance coronary flow during coronary occlusion and reperfusion and in part due to scavenging -OH and/or replenishing intracellular glutathione. The results also indicate that the condition of coronary perfusion can produce a great impact on postischaemic ventricular performance.

An effective method for quantitative evaluation of proteins adsorbed on biomaterial surfaces.

An effective method for the quantitative evaluation of proteins adsorbed on biomaterial surfaces has been developed. First, the kinetic behavior of a range of human fibrinogen (Fib) adsorbed onto polystyrene (PS) films was investigated by using a reflectometry interference spectroscopy setup. The specific molecular number of adsorbed proteins, N(p,) was then defined. According to the definition, the numbers of Fib molecules adsorbed on PS films were calculated. An atomic force microscope (AFM) was used to scan the lateral distribution of the Fib molecules adsorbed on the PS films. From the AFM images, the practical specific molecular numbers were obtained by direct counting of the molecules. In order that the adsorbed number of Fib molecules on a unit area of the PS films could be counted easily, the solution concentration of proteins was reduced to 5 ag/mL (10(-18)g/mL). There was good consistency between the numbers calculated with the formula defined by us and the numbers counted from AFM images. Therefore, the results of the present study prove the validity of our definition of the specific molecular number of adsorbed proteins and the effectiveness of the reflectometry interference spectroscopy-based method for quantitative evaluation of adsorptive proteins.

Neoadjuvant chemotherapy in inoperable, locally advanced, and inflammatory breast carcinoma: a pilot study of MTT assay in vitro and outcome analysis of 10 patients.

Patients with inoperable, locally advanced, and inflammatory breast carcinoma (LAIBC), whether with supraclavicular lymph nodes (SLN) or not (stage IIIB and IV), usually carry an overall poor prognosis. The current treatment for these patients is by means of combined modality, including preoperative chemotherapy. This strategy has led to a substantial improvement in clinical response, making some patients operable, and even making breast conservative surgery possible. However, the long-term results still are not promising. The aim of this pilot study was to determine the efficacy of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay in vitro in directing chemotherapy (including preoperative adjuvant chemotherapy and postoperative adjuvant chemotherapy) for these patients. Between June 1994 and March 1997, 10 patients with inoperable LAIBC, whether with SLN or not, were enrolled. During the period of the combined therapy modalities, the neoadjuvant chemotherapy was adopted for three cycles according to the results of chemosensitivity in vitro by MTT assay. Then a modified radical or radical mastectomy was performed, which was followed by radiotherapy and further postoperative adjuvant chemotherapy with the same regimen as that of neoadjuvant chemotherapy. All patients had been followed up from the beginning of neoadjuvant chemotherapy to the end of October 1999. Two patients had clinical complete response (CRs), with one having pathologic CR in both breast tumor and axillary lymph node, and the other having pathologic CR in axillary lymph node. The other eight patients had partial response. By the time of analysis, six patients had been dead of relapse or progression. Among the four patients who were still alive, one had local relapse, one had distant metastatic disease, and the other two had no evident disease. By retrieving from MEDLINE before 1999, the authors learned that this is the first pilot study of neoadjuvant chemotherapy for inoperable LAIBC using MTT assay to predict the chemosensitivity in vitro. Compared with conventional chemotherapy, the clinical response and long-term results seem to be more encouraging.

[Protective effects of SH-compounds on ischemia reperfusion induced arrhythmias in the isolated rat heart].

Protective effects of cysteine (Cys), N-acetylcysteine (NAC), cysteamine (MEA), cystamine (CSSC) and aminopropylmethylisothiourea (APMT) on ischemia/reperfusion induced arrhythmias were studied in isolated Langendorff perfused rat hearts. The arrhythmias were caused by ligation of the anterior descending branch of the left coronary artery for 10 min and reperfused for 5 min. The drugs were dissolved in saline (NS) and perfused through a peristaltic pump system at 0.1, 0.6 or 3.6 mumol/min (n = 10), starting from 10 min before ligation up to 5 min after reperfusion. The control hearts were perfused with NS. The results showed that Cys, NAC and MEA pursued at 0.6-3.6 mumol/min significantly reduced the incidence of ventricular fibrillation (VF), which were 80-90% in control and 0-20% in 3 treated groups, with P less than 0.01-0.001. The duration of ventricular tachycardia (VT) + VF was 3.0 +/- 1.6 min in control and were 0.2 +/- 0.2, 0.2 +/- 0.1 and 1.2 +/- 2.1 min in Cys, NAC and MEA groups, respectively (with P less than 0.01-0.001). Coronary flow (CF) were remarkably reduced to about 50% during ligation in NS, but remained at normal levels in three treated groups. There were no significant protective effects on arrhythmias in CSSC and APMT perfused hearts. CF of CSSC and APMT groups were even less than those of control. The structure-activity analysis suggested that the SH group may play a crucial role in the protective effect of SH compounds on ischemia/reperfusion induced arrhythmias. The mechanism of protection was briefly discussed in this paper.

["Target secondary structural motif" in the action of antisense oligodeoxynucleotides].

AIM: To optimize the antisense drug design based on the methods of secondary structure prediction of target mRNA by computer and the quantitative structure-activity relationship analysis. METHODS: The secondary structures of mRNA were predicted by the software RNAstructure, then the antisense phosphorothioate oligodeoxynucleotides (AS-ODN) were designed against the secondary structural elements. The in vitro anti-tumor bioactivity of AS-ODN was evaluated by A549 lung carcinoma cell line. The multiple regression was performed with the computer program SPSS. RESULTS: AS-ODN with high bioactivity concentrated on some local secondary structural motifs that were composed of several secondary structural elements, designated here as "target secondary structural motif" (target motif). The target motifs were relatively stable in the whole mRNA structures, but there were one or more unstable secondary structural elements (free energy > 0) such as internal loops, knots, and hairpins, especially the bulge loops in the motif. Bioactivities of AS-ODN targeting different "target motifs" were statistically different (P < 0.01) while AS-ODN against the same "target motif" were with similar effects. CONCLUSION: The concept of the "target motif" was helpful for optimizing the antisense drug design and might be useful for discovering the local function of mRNA and designing oligonucleotide probes and primers.

[Synthesis and properties of 3,17-disubstituted estrogenic steroids].

In order to develop new radio protectors of low estrogenic activity and high potency and to study the structure-activity relationships, 10 estrogenic steroids were synthesized from the view point of drug latentiation, and by substitution with active groups in different positions of the steroidal skeleton. The compounds were tested in mice for estrogenic activity and radioprotective effect. The results showed that the introduction of hydroxyalkylimido or hydroxy group into 17-position resulted in reduction of estrogenic activity. Hydroxyalkylimides (2d, e) showed better protective effect in 750 rad of 60CO gamma-irradiation in mice by the reduction of radiation dose of 144-156 rads than estradiol (reduction of 62.2 rad). Whereas 2a-c and 4a, b showed weak or no protective effect. Compounds 1, 3, and 5a, b increased the 30-day survival rates by 35-80% in mice exposed to 900 rad of irradiation, when administered ip 0.1 mg per mouse 24 h before irradiation.

Chemosensitizing effect of verapamil on Swiss-3T3 cells transfected with human MDR1 gene.

To further understand the characteristics of drug resistance reversal of verapamil, the relationship between the level of doxorubicin resistance and the magnitude of modulation by verapamil was examined in multidrug resistant Swiss-3T3 cells transfected with human MDR1 gene. In independently isolated transfectants doxorubicin cytotoxicity decreased markedly compared with parent cells. Potentiation of doxorubicin toxicity by a noncytotoxic concentration of 3 mumol.L-1 of verapamil was much greater in transfectants than in parent cells, while the magnitude of reversal was inversely dependent on the level of resistance. Southern blot hybridization indicated the MDR1 cDNA integration in genomics of each transfectant. Defect in cellular accumulation of doxorubicin was restored by verapamil in transfected cells. The saturation of active drug transport that may involve the magnitude changes of potentiation by verapamil, and the mode of interaction between P-glycoprotein and drugs, were discussed.

[Therapy of cantharides extract for perennial allergic rhinitis and its effect on total IgE in serum].

The therapy of 10% Cantharides extract in treating 50 cases of perennial allergic rhinitis (PAR) was studied. The extract was plastered and blistered on Dazhui, Neiguan point. It was observed by nasal mucosa provocative test, cells in nasal secretion test and serum total IgE test. The results showed that its effective rate was 88%, the allergic nasal mucosa provocative test of treated group alleviated obviously after the treatment (P < 0.01), the number of eosinophil and basophil in nasal secretion decreased (P < 0.01, P < 0.05); and the serum total IgE also reduced significantly (P < 0.01).

[Clinical study on relationship between liver-blood stasis and liver fibrosis].

OBJECTIVE: Exploring the relationship between Liver-Blood Stasis (LBS) and liver fibrosis (LF) to lay a theoretical foundation of rational using traditional Chinese medicines against LF. METHODS: Human procollagen peptide III (hPC III), hyaluronic acid (HA), laminin (LN) in the sera of 35 patients with hepatopathy and Blood Stasis Syndrome (HP-BS) and 35 patients with hepatopathy and Non-Blood Stasis Syndrome (HP-NBS) were measured by radioimmunoassay. Thirty healthy subjects were taken as control. Correlation analysis between the degrees of LBS and those of the serum indexes of LF was made. RESULTS: (1) hPC III, HA, LN in the sera of the patients with HP-BS were markedly higher than those in the sera of the patients with HP-NBS, but the latter were markedly higher than healthy subjects. (2) Degrees of LBS correlated closely with those of LF. (3) (Xuefu Zhuyu Decoction XFZYD) not only might improve the degrees of LBS but also decline the serum LF indexes. Nevertheless, the curative effects in early period of taking the herbs only showed serum HA dropped. CONCLUSION: The nature of LF in traditional Chinese medicine concept is mainly LBS. The degrees of LBS might reflect those of LF to a certain extent. XFZYD showed effective in declining serum LF indexes but it needs at least 2 months.

Stretched inverse opal colloid crystal substrates-induced orientation of fibroblast.

Recently, there has been increasing interest in studying the interaction between mammalian cells and nanometer-sized structures. However, the effect of nanostructures on cell behavior, such as cell morphology and alignment, is still largely unknown. Inverse opal colloid crystal substrates, which can be stretched to produce nano-scale pore structures of different degrees of orientation, serve as a convenient model system to study the effect of nanotopography on cell morphology and cell alignment. In this work, we fabricated inverse opal colloidal crystal films that were either unstretched or stretched to three, four or six times their original length, producing pore structures of increasing degree of orientation. Human dermal fibroblast-fetal (HDF-f) cells were seeded and cultured on these four types of substrates. The results from fluorescence microscopy and scanning electron microscopy indicated that cells showed the highest degree of alignment when cultured on inverse opal colloid crystal films that were stretched the most (six times original length). The results also demonstrated that the orientation of nanostructures could affect both the morphology and growth direction of fibroblasts. The ability to control the direction of cell growth through the engineering of nanostructures could have important applications in tissue engineering, especially for tissues with anisotropic structures, such as cardiac muscle, blood vessel, tendon and ligament.

[Image analysis of effects of 4 drugs on coronary occlusion and myocardial reperfusion injury].

Myocardial infarction induced by coronary occlusion (O model) and reperfusion (R model) and the effects of nitroglycerin (Nit), propranolol (Pro), lidocaine (Lid), nifedipine (Nif), and saline were studied in rats by image analysis and weighing methods. The results showed that the myocardial infarct size (MIS) in R and O models were 27 +/- 8% and 39 +/- 6%, respectively (P < 0.01). The roundness and the distances from the border of the infarct zone (IZ) to endocardium or epicardium in R model were greater than those in O model, while the gray level difference between normal and infarct myocardium (delta G) in the R model was less than that in O model. MIS of Nit group in O and R models were 23 +/- 12% and 16 +/- 7%, respectively, which were significantly less than those in Lid, Nif groups, and saline. The reduction of MIS was also found in Pro group in both models. The results suggested that early restoration of blood flow resulted in the salvage of injured myocardium and Nit and Pro were found to have beneficial effects in both models.

Pharmacokinetics of recombinant human granulocyte colony-stimulating factor in rabbits and mice.

AIM: To study the pharmacokinetics of recombinant human granulocyte colony-stimulating factor (rhGCSF) in rabbits and mice. METHODS: 125I-rhGCSF was prepared by iodogen method and determined by size exclusive HPLC (SEHPLC). RESULTS: Concentration-time curves after i.v. 125I-rhGCSF in rabbits were best fitted with 2-compartment open model. The alpha and terminal elimination T1/2 were 0.25-0.33 and 3.2-4.6 h, respectively. AUC increased with doses, and Cls and K10 were similar. Tpeak was 0.59 +/- 0.25 h after s.c., and elimination T1/2 was similar to that after i.v. The bioavailability after sc was 1.0. In mice the highest level was found in renal system, the next was bile-enteric system. Levels in lymph nodes, bone marrow, and spleen were approximately equal to or slightly lower than that in plasma, while the levels in brain, fat, and muscles were the lowest. About 68%-86% were recovered in urine and feces. CONCLUSION: Pharamcokinetics of 125I-rhGCSF in rabbits and mice provided a useful index for clinical trial.