Heterozygosity analysis of spontaneous 2n female gametes and centromere mapping of the diploid Hevea brasiliensis based on full-sib triploid populations.

KEY MESSAGE: Unreduced megagametophytes via second-division restitution were confirmed through heterozygosity analysis, and four candidate physical centromeres of rubber were located for the first time. The evaluation of maternal heterozygosity restitution (MHR) is vital in identifying the mechanism of 2n gametogenesis and assessing the utilization value of 2n gametes. In this study, three full-sib triploid populations were employed to evaluate the MHR of 2n female gametes of rubber tree clone GT1 and to confirm their genetic derivation. The 2n female gametes of GT1 were derived from second-division restitution (SDR) and transmitted more than half of the parental heterozygosity. In addition, low recombination frequency markers were developed, and four candidate physical centromeres of rubber tree were located for the first time. The confirmation that 2n female gametes of rubber tree clone GT1 are derived from SDR provides insights into the molecular mechanisms of 2n gametogenesis. In addition, the identified centromere location will aid in the development of centromeric markers for the rapid identification of the 2n gametogenesis mechanism.

Research on the dentification of Panax ginseng and Panax quinquefolium using catalysed hairpin assembly technology.

INTRODUCTION: Panax ginseng and Panax quinquefolium are traditional Chinese herb medicines and similar in morphology and some chemical components but differ in drug properties, so they cannot be mixed. However, the processed products of them are often sold in the form of slices, powder, and capsules, which are difficult to identify by traditional morphological methods. Furthermore, an accurate evaluation of P. ginseng, P. quinquefolium and the processed products have not been conducted. OBJECTIVE: This study aimed to establish a catalysed hairpin assembly (CHA) identification method for authenticating products made from P. ginseng and P. quinquefolium based on single nucleotide polymorphism (SNP) differences. METHOD: By analysing the differences of SNP in internal transcribed spacer 2 (ITS2) in P. ginseng and P. quinquefolium to design CHA-specific hairpins. Establish a sensitive and efficient CHA method that can identify P. ginseng and P. quinquefolium, use the sequencing technology to verify the accuracy of this method in identifying Panax products, and compare this method with high-resolution melting (HRM). RESULTS: The reaction conditions of CHA were as follows: the ratio of forward and reverse primers, 20:1; hairpin concentration, 5 ng/µL. Compared with capillary electrophoresis, this method had good specificity and the limit of detection was 0.5 ng/µL. The result of Panax product identification with CHA method were coincidence with that of the sequencing method; the positive rate of CHA reaction was 100%. CONCLUSION: This research presents an effective identification method for authenticating P. ginseng and P. quinquefolium products, which is helpful to improve the quality of Panax products.

[Functional Genomics Analysis of Nitrogen and Phosphorus Transformation in Maize Rhizosphere Microorganisms].

Fertilizer reduction and efficiency improvement is an important basis for ensuring the safety of the agricultural ecological environment. Microorganisms are the key driving force for regulating the soil nitrogen and phosphorus cycle. Studying the nitrogen and phosphorus transformation function of rhizosphere microorganisms can provide a microbiological regulation approach for further improving the use efficiency of soil nitrogen and phosphorus. Based on the field micro-plot experiments of three typical farmland soils(phaeozem, cambisol, and acrisol), metagenomic sequencing technology was used to study the differences in functional genes and regulatory factors of maize rhizosphere microorganisms during soil nitrogen and phosphorus transformation. The results showed that the functional diversity of maize rhizosphere microorganisms was affected by soil type. The functional diversity of rhizosphere microorganisms in phaeozem and cambisol was mainly affected by water content and nutrient content, and that in acrisol was affected by total phosphorus(TP) and available phosphorus(AP). For soil nitrogen transformation, the gene abundance of related enzymes in the pathway of nitrogen transformation was the highest in the urease gene(ureC) and glucose dehydrogenase gene(gdh), which were 7.25×10-5-12.88×10-5 and 4.47×10-5-7.49×10-5, respectively. The total abundance of assimilatory nitrate reduction functional genes in acrisol was higher than that in phaeozem and cambisol, and the total abundance of functional genes related to other processes was the highest in cambisol. The abundance of functional genes encoding enzymes related to nitrogen metabolism was mainly driven by soil bacterial richness, total potassium(TK), and TP. For soil phosphorus transformation, the number of alkaline phosphatase genes(phoD) catalyzing organic phosphorus mineralization was 1093, and the number of acid phosphatase genes(PHO) was 42. The abundance of phoD was two orders of magnitude higher than that of PHO. In addition, fertilization had no significant effect on the abundance of phoD and PHO in the same soil type. Random forest analysis showed that the abundances of phoD and PHO were significantly affected by soil moisture, organic matter(OM), and total nitrogen(TN), but AP content had the greatest impact on PHO abundance. These results clarified the nitrogen and phosphorus transformation characteristics of maize rhizosphere microorganisms at the functional genomic level and enriched the molecular biological mechanism of the microbial nitrogen and phosphorus transformation function.

Low-carbohydrate diet score and the risk of stroke in older people: Guangzhou Biobank Cohort Study and meta-analysis of cohort studies.

OBJECTIVES: To our knowledge, evidence regarding the association of a low-carbohydrate diet (LCD) with the risk for stroke is limited. The aim of this study was to examine the association between an LCD and stroke in a population with relatively high-carbohydrate intake and risk for stroke and use a meta-analysis to update the evidence. METHODS: The study included 19 850 participants aged ≥50 y without stroke history at baseline in the Guangzhou Biobank Cohort Study. LCD score was calculated based on the percentage of energy from carbohydrate, protein, and fat (range 0-30 points), with a higher score indicating a lower intake of carbohydrate and a higher intake of fat and protein. RESULTS: During an average follow-up of 13.1 y, 1661 stroke events and deaths occurred, including 1255 ischemic strokes. The second, versus the lowest quartile of animal-based LCD scores, were associated with lower risks for all stoke and ischemic stroke (hazards ratio [HR], 0.87; 95% confidence interval [CI], 0.77-0.997, and HR, 0.83; 95% CI, 0.72-0.97). Non-significant associations of plant-based or total LCD scores with the risk for stoke were found. CONCLUSIONS: In a population consuming high levels of carbohydrates and low levels of protein and fat, a moderate level of animal-based LCD, but not plant-based LCD, was associated with a lower risk for stroke.

MRGPRX2 is critical for clozapine induced pseudo-allergic reactions.

BACKGROUND: Clozapine is one of the most widely used second-generation antipsychotics in clinic. However, allergy-like symptoms such as rash and angioedema have been reported frequently, and the mechanism is still not clear. Mas-related G protein-coupled receptor X2 (MRGPRX2) expressed on mast cells is a crucial receptor for drug induced pseudo-allergic reactions. Therefore, we explored whether the symptoms induced by clozapine were associated with allergic reaction through MRGPRX2. METHODS: The effects of clozapine on pseudo-allergic reactions were evaluated by mast cells degranulation and calcium mobilization assay in vitro, and mice hindpaw swelling, serum histamine detection, avidin and H&E staining assay in vivo. The overexpressed MRGPRX2 cells membrane chromatography (MRGPRX2-HEK293/CMC), MRGPRX2-HEK293 cells calcium mobilization assay and molecular docking were applied to research the correlation between clozapine and MRGPRX2. RESULTS: The study showed that clozapine induced the release of ß-hexosaminidase, histamine and monocyte chemoattractant protein-1 (MCP-1), and trigged calcium mobilization in mast cells. In vivo, clozapine induced paw swelling, degranulation and vasodilation. Furthermore, clozapine could activate the calcium mobilization obviously in MRGPRX2-HEK293 cells, not in NC-HEK293 cells. Clozapine also had a good retention characteristic on MRGPRX2-HEK293/CMC column and the K D value is (2.33 ± 0.21)×10-01M. CONCLUSIONS: Our findings demonstrated that clozapine could induce pseudo-allergic reactions and MRGPRX2 might be the critical receptor for it.

Milk consumption and risk of mortality from all-cause, cardiovascular disease and cancer in older people.

BACKGROUND: Milk as a common diet is recommended by many guidelines, but the results on the association of milk consumption with the risk of cardiovascular disease (CVD) or cancer were contradictory. Moreover, evidence regarding milk consumption and mortality risk in Chinese is scarce. OBJECTIVE: We examined the associations of milk consumption with the risk of all-cause, CVD and cancer mortality in a low milk consumption population using data from the Guangzhou Biobank Cohort Study. DESIGN: 18,214 participants aged 50+ years without CVD history at baseline (2003-6) were included. Causes of death were identified through record linkage. Cox proportional hazards regression was used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs). RESULTS: Of the 18,214 participants, 12,670 (69.6%) did not consume milk, 2669 (14.7%) had moderate (1-3 portions/week; 1 portion = 250 ml) and 2875 (15.8%) had high (3+ portions/week) consumption. During an average follow-up of 11.5 (standard deviation = 2.3) years, 2697 deaths occurred, including 917 CVD and 1029 cancer deaths. Compared with no consumption, the adjusted HR (95% CIs) of all-cause, CVD, ischemic heart disease (IHD) and stroke mortality for moderate milk consumption was 0.92 (0.81-1.04), 0.72 (0.57-0.92), 0.57 (0.38-0.85) and 0.77 (0.63-0.94), respectively. High consumption was associated with a higher risk of total cancer and esophagus cancer mortality, with the adjusted HR (95% CIs) being 1.33 (1.12-1.57) and 3.20 (1.21-8.43) respectively. No significant association of high consumption with lung cancer, liver cancer, gastrointestinal cancer, or colorectal and anal cancer was found. CONCLUSIONS: In our sample of Chinese with much lower milk consumption than those in the West, compared with no consumption, moderate milk consumption showed a lower risk of CVD mortality, but high milk consumption showed a higher risk of total cancer mortality. Further studies are warranted to verify the differential effects of milk on CVD and cancer.

SOX2-induced upregulation of lncRNA LINC01510 promotes papillary thyroid carcinoma progression by modulating miR-335/SHH and activating Hedgehog pathway.

LncRNA LINC01510 (LINC01510) was a newly identified tumor-related lncRNA whose dysregulation and potential function have been reported in several tumors. However, the expression, clinical significances, and action mechanisms of LINC01510 in papillary thyroid carcinoma (PTC) are still unclear. In this study, we firstly reported that LINC01510 was highly expressed in both PTC tissues and cell lines. Additionally, we used dual-luciferase reporter assay and confirmed that SOX2 could bind directly to the LINC01510 promoter region, activating its transcription. Functional assays with a series of cell experiments indicated that knockdown of LINC01510 suppressed the proliferation, migration and invasion of SW1736 and TPC-1 cells. Moreover, down-regulation of LINC01510 resulted in accelerated apoptosis by promoting the expression of Caspase3/9. In particular, LINC01510 acted as an endogenous sponge by directly binding miR-335, resulting in the suppression of miR-335 expressions. Besides, we confirmed that SHH was a target of miR-335 and miR-335 over-expression distinctly reduced SHH expression in PTC cells. Finally, in the cytoplasm, we provided evidenced that LINC01510 acted as a sponge for miR-335, reducing its ability to promote SHH expression. In addition, the results of Western blot indicated that knockdown of LINC01510 inhibited the expression of SHH and GLI1, suggesting that Hedgehog pathway was suppressed. Taken together, our findings revealed that the newly identified LINC01510/miR-335/SHH axis could be a therapeutic target for PTC.

Expression of recombinant human acetylcholinesterase and its application in screening its inhibitors.

This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.

Breast cancer in Iraq, incidence trends from 2000-2009.

BACKGROUND: Breast cancer is the most frequent malignancy of women worldwide. In Iraq, breast cancer ranks first among cancers diagnosed in women but no studies have been conducted on incidence trends. The present study of breast cancer in the country during 2000-2009 was therefore performed. MATERIALS AND METHODS: The registered data for breast cancer cases were collected from the Iraqi Cancer Registry/Ministry of Health. The significance of incidence rate trends during 2000-2009 was tested using Poisson regression. Age-standardized rates (ASR), and age-specific rates per 100,000 population were calculated. RESULTS: A total of 23,792 incident breast cancer cases were registered among females aged ≥15 years, represented 33.8% of all cancers in females registered during 2000-2009. It ranked first in all the years. The median age at diagnosis was 49 and the mean age was 52 years. The incidence rate of all female breast cancer in Iraq (all ages) increased from 26.6 per 100,000 in 2000 to 31.5 per 100,000 in 2009 (APC=1.14%, p<.0001). The incidence in age groups (40-49), (50-59) and (70+) increased in earlier years and has recently (2005-2009) become stable. The incidence in age group (60-69) did not decline since 2003, while the incidence rates in the age group (15-39) started to decline in 2004. CONCLUSIONS: With the Iraqi Cancer Registry data during the period 2000-2009, the incidence of all female breast cancer in Iraq (all ages) has risen. We found rapid increase in the age specific incidence rate among age group 60-69. However, breast cancer among Iraqi women still affects younger age groups than their counterparts in developed countries. Further epidemiological research is needed to examine possible causes and prevention measures.

Trend analysis of lung cancer incidence rates in ninawa province, Iraq, from 2000 to 2010--decrease and recent stability.

BACKGROUND: Lung cancer is the most frequent malignancy of men worldwide. In Ninawa in Iraq, lung cancer ranks first among cancers diagnosed in men. Since no prior studies have been conducted on incidence trends in our population the present investigation of rates during 2000-2010 was therefore performed. MATERIALS AND METHODS: Registy data for lung cancer cases were collected from the Directorate of Health in Ninawa-Mosul Continuing Medical Education Center. We restricted our analyses to men categorized according to the age groups of 0- 39, 40-49, 50-59, 60-69 and 70+ years. The significance of incidence rate trends during 2000-2010 was tested using Poisson regression. Age-standardized rates (ASR), and age-specific rates per 100,000 population were calculated. RESULTS: A total of 1,206 incident lung cancer were registered among males, accounting for 15.5% of all male cancers registered during 2000-2010. It ranked first throughout the period. Median age at diagnosis was 69 (mean 66.8± 11.0) years. The incidence rate of all male lung cancers in Ninawa (all ages) decreased from 26.4 per 100,000 in 2000 to 12.7 in 2010 (APC=-6.55%, p<.0001). The incidences in age groups 40-49, 50-59, 60-69 and 70+ decreased in earlier years and recently appeared (2007-2010) stable. The incidence in age group (0-39) remained stable between 2000-2010. Squamous cell carcinoma (SCC)was the most common type of lung cancer, while adenocarcinoma was relatively rare. CONCLUSIONS: With the data from Directorate of Health in Ninawa during the period 2000-2010, lung cancer is the most common cancer but generally declining. Among all age groups, the recent incidence of lung cancer remained stable. The SCC predominance suggests change in tobacco habits as an important factor in the trends observed.

ADAM17 mediates hypoxia-induced drug resistance in hepatocellular carcinoma cells through activation of EGFR/PI3K/Akt pathway.

A disintegrin and metalloproteinase-17 (ADAM17) is a member of the metalloproteinase superfamily and involved in the cleavage of ectodomain of many transmembrane proteins. ADAM17 is overexpressed in a variety of human tumors, which is associated with tumor development and progression. In the present study, we sought to investigate the expression and function of ADAM17 in hypoxia-treated hepatocellular carcinoma (HCC) cells. Western blot analysis was used to measure the expression of ADAM17 in HCC cell lines (Hep3B and HepG2 cells). Annexin V/PI double staining was performed to analyze the effects of ADAM17 on hypoxia-mediated cisplatin resistance. ADAM17 expression was upregulated by hypoxia treatment in HCC cells at both mRNA and protein levels. Overexpression of ADAM17 reduced cisplatin-induced apoptosis in HCC cells, accompanies by less cleavage of caspase-3 and poly (ADP-ribose) polymerase (PARP). Forced expression of ADAM17 enhanced the phosphorylation of epidermal growth factor receptor (EGFR) and Akt without affecting the expression of total EGFR and Akt. Pretreatment with EGFR inhibitor AG1478 or phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 rescued ADAM17-mediated cisplatin resistance of HCC cells. ADAM17 silencing attenuated hypoxia-induced cisplatin resistance and enhanced the accumulation of cleaved caspase-3 and PARP. Western blot analysis showed that overexpression of hypoxia-inducible factor-1α (HIF-1α), a transcription factor, upregulated the expression of ADAM17 and HIF-1α silencing downregulated the expression of ADAM17 in hypoxia-treated HCC cells, indicating the regulation of ADAM17 by HIF-1α. Taken together, our results indicated that ADAM17 is upregulated by hypoxia and contributes to hypoxia-induced cisplatin resistance via EGFR/PI3K/Akt pathway.

[Pollution characteristics of microbial aerosols generated from a municipal sewage treatment plant].

To characterize the pollution characteristics of microbial aerosols emitted from municipal sewage treatment plants, microbial aerosols were sampled with an Andersen 6-stage impactor at different treatment units of a Xi'an sewage treatment plant between June 2011 and July 2011. The plate-culture and colony-counting methods were employed to determine the concentrations, particle size distributions and median diameters of the airborne bacteria, fungi and actinomycetes. The results showed that the highest concentrations of bacteria (7 866 CFU x m(-3) +/- 960 CFU x m(-3)) and actinomycetes (2 139 CFU x m(-3) +/- 227 CFU x m(-3)) were found in the sludge-dewatering house while the highest fungi concentration (2156 CFU x m(-3) +/- 119 CFU x m(-3)) in the oxidation ditch. The airborne bacteria, fungi and actinomycetes all showed a skewed distribution in particle size. The peaks of bacteria and fungi were in the size range of 2.1-3.3 microm, whereas the peak of airborne actinomycetes was between 1. 1-2.1 microm in size. In general, the order of the median diameters of different microbial aerosols generated from the sewage treatment plant was airborne bacteria > airborne fungi > airborne actinomycetes. In addition, the spatial variation characteristics of microbial aerosols showed that the larger the particle size of the microorganism, the faster the reducing rate of the aerosol concentration. The variations in the reducing rate of concentration with particle sizes can be ordered as airborne bacteria > airborne fungi > airborne actinomycetes.

Pharmacokinetics and plasma protein binding of rutin deca (H-) sulfate sodium.

Rutin deca (H-) sulfate sodium (RDS) possesses very good activity as an inhibitor of the complement system of warm-blooded animals and HIV. An ion-pair coupled with solid-phase extraction technique (IP-SPE) was developed to extract RDS from rat plasma, urine, bile and protein solution samples. The assay was applied to pharmacokinetics of RDS, including plasma pharmacokinetics, excretion and protein binding studies. After i.v. 5, 20 and 100 mg x kg(-1) RDS via tail vein in rats, the plasma concentration-time profiles were fitted using 3P97 software. The average terminal half-life (t(1/2)) was 3.432 +/- 0.185 2 h. The relationship of dose and AUC of RDS was linear within the dosage range. This suggested that the disposition of RDS in rats belong to linear kinetics and the pharmacokinetic parameters of RDS were dose independent. After iv RDS 20 mg x kg(-1) in rats, the biliary excretion amount of parent drug amount was only 0.3181% +/- 0.2087% of given dosage, and the urinary excretion was 86.0% +/- 6.1% in 36 h. Ultrafiltration techniques were applied to determine the protein binding of RDS in plasma (from SD rat, Beagle dog and human), human serum albumin (HSA) and human alpha1-acid glycoprotein (AGP). The mean protein binding rate in plasma of SD rat, Beagle dog and human plasma of RDS were 80%-90%, in which the range of concentration of RDS was 5 to 100 microg x mL(-1). The protein binding to HSA was 85.7% +/- 1.3% and 14.0% +/- 3.2% to AGP.

Simultaneous determination of mandelic acid enantiomers and phenylglyoxylic acid in urine by high-performance liquid chromatography with precolumn derivatization.

A reversed-phase HPLC method for the simultaneous quantitative determination of mandelic acid enantiomers (MA) and phenylglyoxylic acid (PGA) in urine is described. MA and PGA were extracted with ethyl acetate from urine at acidic pH and derivatized with S-(-)-1-(1-naphthyl) ethylamine. A ZORBAX SB-C(18) column (250 mm x 4.6mm i.d., 5 microm, Agilent, USA) was used with a mobile phase composed of methanol-10 mmol/L phosphate buffer [pH 2.5 (65:35, v/v)] at a flow-rate of 0.8 ml/min. Detection was set at UV wavelength of 254 nm. The mean absolute recoveries were 94.2%, 91.9%, 92.5% and 86.3% for S-MA, R-MA, PGA and salicylic acid (I.S.), respectively. The intra- and inter-day precisions determined at three different concentrations ranged from 2.8% to 4.8%, 0.7% to 7.7% and 1.3% to 6.8%, respectively. The lower limits of detection for MA enantiomers and PGA in urine were 1 microg/ml and the lower limits of quantification were 5 microg/ml (R.S.D.<10%, n=5). The method has been applied to determine the urinary excretion of MA enantiomers and PGA from Sprague-Dawley rats after orally administered with styrene.

Determination of rutin deca(H-) sulfate sodium in rat plasma using ion-pairing liquid chromatography after ion-pairing solid-phase extraction.

Rutin deca(H-) sulfate sodium (RDS) is one of the most important drug candidates, which possesses very good activity as inhibitor of the complement system of warm-blooded animals and human immunodeficiency virus (HIV). In order to understand RDS metabolism and disposition, an ion-pairing coupled with solid-phase extraction technique (IP-SPE) was developed to extract RDS from rat plasma sample. Tetrabutyl ammonium bromide (TBAB) buffer (0.2 M, pH 8.0) was used as the ion-pairing extraction reagent and LC-18 was used as SPE sorbent. In addition, an ion-pairing HPLC method was established for the specific determination of RDS. A reversed phase C8 column was used for the separation of RDS and nitrendipine (internal standard). The mobile phase was composed of 10 mM phosphate buffer solution containing 25 mM TBAB-acetonitrile (52:48, v/v, pH 7.5). The calibration curve was linear from 0.3 to 30 nmol/mL. The analytical recovery from rat plasma was found to be 97.9+/-4.1% (n = 15). LOD and LOQ for RDS in plasma were calculated to be 0.12 nmol/mL and 0.30+/-0.024 nmol/mL (R.S.D. = 8.2%, n = 5), respectively. The intra- and inter-day precision was less than 9.2%. The assay was applied to a preliminary pharmacokinetic study in three male rats after those received a single intravenous bolus via caudal vein of 12 micromol/kg RDS.

Separation of rutin nona(H-) and deca(H-) sulfonate sodium by ion-pairing reversed-phase liquid chromatography.

Ion-pairing reversed-phase liquid chromatography (RPLC) was used to separate two polysulfonates, rutin nona(H-) sulfonate sodium and rutin deca(H-) sulfonate sodium, which have very similar chemical structures. The final product always contained both of them when one of the compounds was synthesized. Baseline separation was achieved on a C8-bonded silica column at ambient temperature. The eluent was acetonitrile-15 mM phosphate buffer solution containing 20 mM TBA (pH 6.0) (46:54, v/v). The calibration plot was linear in the concentration range 0.5-200 microg ml(-1) for both analytes. The limits of detection (LODs; 254 nm) were 0.03 microg ml(1-) for rutin nona(H-) sulfonate sodium and 0.04 microg ml(-1) for rutin deca(H-) sulfonate sodium. Three batches of rutin deca(H-) sulfonate sodium were analyzed using the assay; the results showed that the analytical performance is really satisfactory.

[HPLC determination of zolmitriptan and its related substances].

OBJECTIVE: To develop an analytical method and quality control for determination of zolmitriptan and related substances. METHODS: Zolmitriptan and related substances were separated and determined on a shimadzu CLC-C(8) column (150 mm x 6 mm, 10 microm) with a mobile phase of acetonitrile-10 mmol/L phosphate buffer (25:75 pH 7.5) and a flow-rate of 1 ml/min; the UV-VIS detector was operated at 229 nm. RESULT: The limit of detection for the related substances was 0.5 ng on the zolmitriptan basis (S/N >3). Linear calibration curve was gene rated from 4 - 40 microg/ml with a correlation coefficient of 0.9999. The recovery rate of zolmitriptan was 99.1% with a standard deviation of 0.2%. The results of HPLC method were consistent with those of nonaqueous titration method. CONCLUSION: HPLC method is a rapid sensitive and accurate method for the determination of zolmitriptan and its related substances.

[Determination of three formulations of pilocarpine in rabbit ocular aqueous by RP-HPLC].

OBJECTIVE: To develop an RP-HPLC method for assay of pilocarpine in rabbit ocular aqueous humor. METHODS: The RP-HPLC method was performed on a column of ODS-C(18) with the mobile phase consisting of 0.5% of triethylamine (TEA) of phosphate solutions (10 mmol/L, pH 2.5) and acetonitrile (98/2,v/v). The detection wavelength was 215 nm and flow rate was 1.0 ml/min. Ninety albino rabbits were divided into 3 groups (30 in each):group 1 received 50 microl of eye drops containing 1% generic pilocarpine, group 21% mixture pilocarpine solution consisting of aqueous sample and liposome and group 31% liposome pilocarpine, respectively. The aqueous humor was withdrawn at 5, 10, 30, 40, 60, 90, 120, 180, 240 and 360 min. Pilocarpine was extracted from aqueous humor with dichloromethane. RESULT: The linear calibration curve was obtained in the concentration range of 0.1 - 20 microg/ml. The average recovery was (68.1+/-2.7)% (n=9). Inter-day and intra-day RSD were 4.33% and 2.87%, respectively. In three formations 1% liposome pilocarpine was the best for the areas under curve and measurable amounts. CONCLUSION: The RP-HPLC method is simple and reliable for pilocarpine measurement in ocular aqueous. Liposome formulation can significantly increase the bioavailability of pilocarpine in ocular aqueous.

[Enatiomeric separation of beta-blocking agents and analogs]

OBJECTIVE: To evaluate enantiomeric separation methods for beta-blocking agents and analogs. METHODS: Enantiomeric separation of racemates of 11 beta-blocking agents and their analogs was performed using chiral stationary phases and 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate (GITC). RESULTS: These beta -blocker racemates were separated into enantiomers in one or several chormatographic states such as propranolol, bisoprolol, metoprolol, celiprolol, carvedilol, sotalol, propafenone, ephedrine, and zomitriptan. Temperature had a significant effect on the resolution of the drugs when using chiralcel OD. Lower temperatures were associated with higher resolutions. CONCLUSION: When separating beta-blocking agents and their analogs, Chiralcel OD, Chiralpak AD, Chiral stationary phases and GITC chiral derivative reagents have complementary functions.