Rational design and synthesis of triazene-amonafide derivatives as novel potential antitumor agents causing oxidative damage towards DNA through intercalation mode.

In this work, we rationally designed and synthesized two novel triazene-amonafide derivatives 2-(2-(diisopropylamino)ethyl)-5-(3,3-dimethyltriaz-1-en-1-yl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (D-11) and 5-(3,3-diethyltriaz-1-en-1-yl)-2-(2-(diisopropylamino)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (D-12) as potential antitumor agents. The DNA damage induced by the intercalation mode of D-11 (D-12) towards DNA was electrochemically detected through the construction of efficient biosensors. The consecutive processes of reversible redox of naphthylimide ring and irreversible oxidation of triazene moiety were elucidated on the surface of glassy carbon electrode (GCE) by CV, SWV, and DPV methods. Electrochemical biosensors were obtained through the immobilization of ctDNA, G-quadruplexes, poly(dG), and poly(dA), respectively, on the clean surface of GCE. After the incubation of biosensors with D-11 or D-12, the peaks of dGuo and dAdo decreased prominently, and the peak of 8-oxoGua appeared at +0.50 V, suggesting that the interaction between D-11 (D-12) and DNA could result in the oxidative damage of guanine. Unexpected, the as-prepared DNA biosensor possessed satisfactory anti-interference property and good practicability in real samples. UV-vis and fluorescence spectra, and gel electrophoresis assays were employed to further confirm the intercalation mode of D-11 (D-12) towards DNA base pairs. Moreover, D-11 was proved to exhibit stronger anti-proliferation activity than mitionafide and amonafide against both A549 and HeLa cell lines.

Synthesis and biological evaluation of aminobenzamides containing purine moiety as class I histone deacetylases inhibitors.

The aminobenzamide is selective to class I histone deacetylases (HDACs) and displays unique tight-binding/slow-off HDAC-binding mechanism. Herein, we report a series of 9-substituted purine aminobenzamides that selectively inhibit class I HDACs. The activities in vitro showed compound 9d exhibited 12 folds more potent than MS-275 against HDAC1 isoform and showed excellent inhibitory activity on cancer cells, including HCT-116, MDA-MB-231, K562 cell lines. The metabolic stability of 9d was much better than that of the well-known HDAC inhibitor SAHA. Pulse exposure test of western blot assay demonstrated that 9a, 9d induced histone acetylation in a similar manner to MS-275. Further biological validation demonstrated that 9d prevented cell transition from G1 phase to S phase by reducing Cyclin D1, CDK2 and lifting p21, induced early apoptosis by upregulating BAX and downregulating Bcl-2 in HCT-116 cells.

[The impurity profiling of paclitaxel and its injection by UPLC-MS/MS].

An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method was developed to elucidate the impurity profiles of paclitaxel and paclitaxel injections from different Chinese pharmaceutical companies. The fragmentation patterns for paclitaxel and the related impurities were analyzed and summarized. To remove the interference from auxiliary materials, such as hydrogenated castor oil, paclitaxel was dissolved in ethanol for acid, base, peroxide, and light induced forced degradation analysis, which could produce all the impurities exist in the paclitaxel injection. A total of 10 impurities were characterized, such as cephalomannine (1), 7-epi-10-deacetylpaclitaxel (2), 7-epipaclitaxel (3), baccatin Ⅲ (4), ethyl ester side chain (5), 7-epi-baccatin Ⅲ (6), 10-deacetylpaclitaxel (7), paclitaxel isomer(C3-C11 bridge) (8), paclitaxel isomer (9), and N-benzoyl-(2R,3S)-3-phenylisoserine (10), respectively. Among them, compounds 1-3 could be introduced during manufacture processing. In the forced degradation studies, while acid induced degradation products included 3-7, base induced degradation could produce 2-7 and 10; while 7 is the main compound produced by hydrogen peroxide treatment, 4 compounds (3-5 and 7) were produced by high temperature environment and 5 compounds (2-5 and 9 which is the first reported) from intensity light exposure. Furthermore, 8 was the main impurity came from intensity light exposed paclitaxel powder. The results from this study provide an important reference in processing, optimization, quality control and evaluation of paclitaxel.

First provincial survey of Angiostrongylus cantonensis in Guangdong Province, China.

The rat lungworm Angiostrongylus cantonensis is a zoonotic nematode with a wide distribution. We report the first provincial survey of the prevalence of A. cantonensis infection among wild rodents and snails in Guangdong Province, China. A total of 2929 Pomacea canaliculata and 1354 Achatina fulica were collected from fields in 22 survey sites with a larval infection rates ranging from 0-26.6% to 0-45.4%. In addition, 114 Cipangopaludina sp and 252 Bellamya sp were bought from markets; larvae were found only in Bellamya snails from two survey sites with an infection rate of 1.4% (1/70) and 3.3% (3/91), respectively. Four hundred and ninety-one rodents were captured in nine sites (Rattus norvegicus, R. flavipectus, Suncus murinus, Mus musculus, Bandicota indica, R. losea and R. rattus). Adult worms were found in R. norvegicus, R. flavipectus and Bandicota indica. Our survey revealed a wide distribution of A. cantonensis and its intermediate hosts P. canaliculata and A. fulica in Guangdong. The prevalence of A. cantonensis in wild snails and rats poses a substantial risk for angiostrongyliasis in humans.

Aluminium-phosphorus interactions in plants growing on acid soils: does phosphorus always alleviate aluminium toxicity?

Aluminium (Al) toxicity and phosphorus (P) deficiency are considered to be the main constraints for crop production in acid soils, which are widely distributed in tropical and subtropical regions. Conventionally, P addition is regarded as capable of alleviating Al toxicity in plants. However, this field is still rife with unsubstantiated theories, especially for different plant species growing on acid soils. In this review, the responses of plants to different methods of Al-P treatments are briefly summarized, and possible reasons are proposed by considering recent results from our laboratory. It is shown that: (1) long-term Al-P alternate treatment is advantageous for studying Al-P interactions in plants; (2) under the long-term Al-P alternate treatment, the roles of P in Al phytotoxicity might be associated with the Al resistance capability and P use efficiency of the plant, and a P/Al molar ratio exceeding 5 in roots may be the threshold of P alleviating Al toxicity based on the calculation of the tested plants; (3) in acid soils, P application may be effective only after Al stress is overcome for Al-sensitive species. Thus it is concluded that P application does not always alleviate Al toxicity under long-term Al-P alternate treatment.

[Methods for grain size analysis of nanomedicines].

As nanomedicines are developing fast in both academic and market areas, building up suitable methods for nanomedicine analysis with proper techniques is an important subject, requiring further research. The techniques, which could be employed for grain size analysis of nanomedicines, were reviewed. Several key techniques were discussed with their principles, scope of applications, advantages and defects. Their applications to nanomedine analysis were discussed according to the properties of different nanomedicines, with the purpose of providing some suggestions for the control and administration of nanomedicines.

[Analysis and evaluation of the impurity of felodipine and its tablets].

The paper reports the systematic study on felodipine and its impurities in tablets, to improve its quality standards for the control of the related substances. HPLC-DAD, UPLC-MS, IR and NMR methods were used for the isolation of felodipine and its impurities in tablets, their identification and the zebrafish animal model was used for the analysis of the toxic impurities. In felodipine material and its tablets, three impurities are isolated and identified. They are impurity 1 [dimethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], impurity 2 [ethyl methyl 4-(2, 3-dichlorophenyl)-2, 6-dimethylpyridine-3, 5-dicarboxylate] and impurity 3 [diethyl 4-(2, 3-dichlorophenyl)-2, 6-dimethyl-1, 4-dihydropyridine-3, 5-dicarboxylate], separately. The result of zebrafish animal model analysis showed that the teratogenic effects of four compounds were: impurity 3 > or = felodipine > impurity 1 > impurity 2, lethal effects were as follows: impurity 2 = impurity 3 > felodipine > or = impurity 1. This study confirmed the toxicity of three impurities in felodipine. According to the results, the paper suggested the amendments to the standard of the medicine and provided the support to the control of impurities in the manufacturing process.

The TCM pattern of the six-zang and six-fu organs can be simplified into the pattern of five-zang and one-fu organs.

OBJECTIVE: To simplify the TCM pattern of the zang-fu organs. METHODS: A database of zang-fu syndromes was established. The relationship between the zang-fu syndromes was analyzed by means of frequency analysis, association rule, and phi-coefficient correlation. RESULTS: The six zang-fu organs of the heart, liver, spleen, lung, kidney, and stomach prove to be the main components of the zang-fu pattern. The large intestine, small intestine, gallbladder, bladder, triple energizer, and pericardium have strong association and positive correlation with the above-mentioned six zang-fu organs, and their clinical symptoms can all be included into the symptoms of these six zang-fu organs. CONCLUSION: The traditional TCM pattern of six-zang and six-fu can be simplified into five-zang and one-fu.

[Investigation on Angiostrongylus cantonensis infection in rodents in Guangdong Province].

OBJECTIVE: To investigate the natural infection of Angiostrongylus cantonensis in Guangdong Province, and to provide the scientific evidence for control measures. METHODS: The investigation was carried out in 56 villages of 28 towns of 28 counties/districts in East Guangdong, West Guangdong, the mountain area of North Guangdong and Peal River Delta of the Province from 2005 to 2010. The rodents were captured with live trap and the species identified. Angiostrongylus cantonensis adult worms were collected from the hearts and lungs of rodents, examined, counted and the sex of worms identified. RESULTS: The rodents were captured from 2005 to 2010, belonged to 2 orders, 2 families (subfamily), 4 genera and 10 species. Seven species of the rodents were found infected with Angiostrongylus cantonensis in all 28 counties/districts. Totally 5 820 rats were examined and 496 infected ones were identified, with a mean infection rate of 8.52%. The infection rate of rodents was highest in the Peal River Delta, reaching 9.8% (205/2084) (chi2=15.25, P<0.01). Rattus norvegicus had the highest infection rate of 16.9% (310/1 835) (chi2=240.91, P<0.01). The mean intensity of infection was 6.1 worms/rat. 1 125 female and 1064 male worms were found respectively (chi2=1.75, P>0.05). CONCLUSION: Natural infection of Angiostrongylus cantonensis in rodents has been found in all the 56 villages selected from the 4 regions of Guangdong Province.

Development and validation of HPLC-UV-MS method for the control of four anti-diabetic drugs in suspected counterfeit products.

An HPLC-UV method has been developed for the determination of valibose, miglitol, voglibose and acarbose, the four anti-diabetic drugs. The separation was accomplished successfully by using reversed phase chromatography (Prevail carbohydrate column, 250 mm x 4.6 mm, 5 microm) with a gradient acetonitrile-phosphate buffer solution (pH 8.0) at a wavelength of 210 nm. Furthermore, the method of a high-performance liquid chromatography coupled with ESI-MS in positive ionization mode has been established. These two methods were successfully applied to the assay and qualitative detection of four alpha-glucosidase inhibitors in the potential counterfeit anti-diabetic drugs.

Nineteen clinical features of fever in Chinese medicine.

Based on the 4,067 descriptions about fever in the medical records of the well-known Chinese medicine physicians throughout the ages, 19 different clinical features of fever are summarized as follows: chilly fever, high fever, low fever, dull fever, bones fever, alternating chills and fever, tidal fever, head fever, face fever, chest fever, back fever, abdomen fever, waist fever, palms and soles fever, lower extremity fever, afternoon fever, evening fever, postpartum fever, and menstrual fever. Although, some of these fever features have not be paid close attention to in Western medicine, stage-division and classification about fever in Western medicine deserve to be used by Chinese medicine for reference.

[Raman spectroscopy fluorescence background correction and its application in clustering analysis of medicines].

During Raman spectroscopy analysis, the organic molecules and contaminations will obscure or swamp Raman signals. The present study starts from Raman spectra of prednisone acetate tablets and glibenclamide tables, which are acquired from the BWTek i-Raman spectrometer. The background is corrected by R package baselineWavelet. Then principle component analysis and random forests are used to perform clustering analysis. Through analyzing the Raman spectra of two medicines, the accurate and validity of this background-correction algorithm is checked and the influences of fluorescence background on Raman spectra clustering analysis is discussed. Thus, it is concluded that it is important to correct fluorescence background for further analysis, and an effective background correction solution is provided for clustering or other analysis.

[Fingerprints of Curcuma wenyujin by RP-HPLC].

OBJECTIVE: To establish a mutual mode fingerprint of Curcuma wenyujin for quality control of C. wenyujin. METHODS: Waters Symmetry C18 column was used; the mobile phase was methanol-water in a linear gradient elution with the flow rate of 1.0 mL/ min, the temperature of column was 30 degrees C; the detection wavelength was set at 215 nm. 10 batches of C. wenyujin from different places and different time were determined, 3 batches of C. phaeocaulis and 3 batches of C. kwangsiensis were determined in the same chromatographic conditions. RESULTS: 11 mutual peaks in the fingerprint of the 10 groups of C. wenyujin, and the S peak among them represented germacrone. CONCLUSION: Curcuma wenyujin's fingerprints have strong feature and specificity, which can be combined with assaying in the quality control of C. wenyujin.

[Survey on the focus of Angiostrongylus cantonensis in Guangdong Province].

OBJECTIVE: To understand the distribution of Angiostrongylus cantonensis foci in Guangdong Province for making surveillance program. METHODS: Survey sites were chosen by strata sampling according to different geographic locations. Totally 22 survey sites were selected in four regions: East Guangdong, West Guangdong, North Guangdong and the Pearl River Delta. One or two administrative villages in each site were randomly selected for the investigation. Pomacea canaliculata and Achatina fulica collected from fields and other species of freshwater or terrestrial snails obtained in the restaurants and wet markets were examined for the third stage larvae by tissue grinding or lung examination. Rats were captured in the fields, and their hearts and lungs were dissected for adult worms. Rat feces were also collected for the detection of first stage larvae by water precipitation. RESULTS: Large number of P. canaliculata was found in all sites. A. fulica was found in most surveyed sites. Totally 2929 P. canaliculata and 1354 A. fulica were collected with a larva infection rate of 5.9% (172/2929) and 16.5% (223/1354), respectively (P<0.01). The average prevalence among the regions was different (P<0.01) with the highest prevalence in Pearl River Delta (15.6%, 152/975), especially in Dongguan City of the Delta (34.7%, 78/225). 114 Cipangopaludina sp. and 252 Bellamya sp. were bought from wet markets of 9. sites. Larvae were found only in Bellamya snails from Luoding and Kaiping cities with an infection rate of 1.4% (1/70) and 3.3% (3/91), respectively. Totally 491 rats were captured in 9 sites including Rattus norvegicus, R. flavipectus, Suncus murinus, Mus musculus, Bandicota indica, R. losea and R. rattus, with an average infection rate of 11.4% (56/491). Adult worms were found in R. norvegicus, R. flavipectus and B. indica with a prevalence of 19.8% (52/263), 2.5% (3/118) and 10.0% (1/10), respectively. Thirty-four rodent fecal samples were collected in 7 sites and examined with a larva positive rate of 44.1% (15/34). CONCLUSION: Foci of Angiostrongylus cantonensis are widely distributed in Guangdong Province as natural infection has been found in its intermediate and definitive hosts.

Research on Treatment of Oily Sludge from the Tank Bottom by Ball Milling Combined with Ozone-Catalyzed Oxidation.

Difficult separation of oil-solid phase and high fine content of the recovered oil were two problems in the treatment of oily sludge from the tank bottom by the hot water-based extraction process. To solve the problems, one technology with "ball milling + ozone-catalyzed oxidation" as the core was studied, and the process parameters of ball milling and ozone-catalyzed oxidation were respectively optimized. After ball milling treatment, the oil content of dry oily sludge decreased from 33.9 to 10.2%. Then, an ozone catalytic oxidation treatment technology with aluminum ore as the catalyst was developed to further treat this stubborn oily sludge. Under the optimal conditions, the oil content of oily sludge could be further reduced to 0.28%, which met the treatment and disposal requirements stipulated in GB4284-2018. For further research on the contribution of the catalyst to the ozone catalytic oxidation system, the reaction activation energy and reaction rates of ozone oxidation and ozone catalytic oxidation were compared from the perspective of kinetics. The results showed that, with the catalyst addition, the reaction rate constants increased about three times and the reaction activation energy reduced 82.26%, which showed the effectiveness of the catalyst on the kinetics quantitatively. The combined process with "ball milling + ozone-catalyzed oxidation" as the core can solve the two problems in the treatment of oily sludge from the tank bottom by hot water-based extraction and provides a reference for the harmless and resourceful treatment of oily sludge from the tank bottom.

The control of clonorchiasis in Guangdong province, southern China.

Oriental liver fluke disease (clonorchiasis), caused by Clonorchis sinensis, is endemic in 81 counties in Guangdong province, China. This study was conducted following increasing lack of clonorchiasis awareness and increased consumption of raw freshwater fish and raw fish porridge at home and restaurants, which has been attributed to improved living standards in China, and is considered to have led to a higher C. sinensis infection rate in the country. A total of 61,517 individuals were investigated from 123 sites during a provincial survey in 1988, with average C. sinensis infection rate of 1.82% that increased to 4.08% (335/8217) in 1997. Higher infection rate was observed in adult men than women, with heavy infection rate in fishermen and businessmen because they have access and means to get raw fishes for consumption. Thus, clonorchiasis has become a serious public health problem in Guangdong province. Efforts targeting clonorchiasis prevention and control in Guangdong province started in the 1990s. These included carrying out large-scale surveys and chemotherapy in epidemic regions, research on promotion of environmental sanitation and methods for freshwater aquaculture. Establishment and promotion of clonorchiosis demonstration plots for comprehensive control measures, as well as the development of clonorchiasis-prevention community-out-patient clinics were initiated and their effectiveness was evaluated. However, there seem to be no obvious decline in the prevalence of C. sinensis with 4.90% (608/12,401) in 2015, at Guangdong province due to the people's habit of eating raw fishes in endemic areas. This has led to increasing calls to improve environmental sanitation in the aquaculture industry. It is, however, believed that control efforts on clonorchiosis in the province are at an early stage and require strengthening through the cooperation of different departments to work out effective strategies for sustainable field application of control mechanisms.

Establishment of an HPLC identification system for detection of counterfeit steroidal drugs.

A set of simple HPLC methods employing UV detection were developed for detection of counterfeit drugs by the qualitative and quantitative analysis of nine steroidal drugs, ethinylestradiol, diethylstilbestrol, norethisterone, norgestrel, methyltestosterone, medroxyprogesterone acetate, progesterone, testosterone propionate and nilestriol. The methods were based on studies of the relationships between the retention factors (k) of the nine compounds and the percentages of water to methanol in the mobile phases on a reverse phase Alltima C(18) column giving reliable separation of the compounds under three sets of chromatographic conditions. The methods were validated using statistical tests and were used on nine commercial samples for detection of possible counterfeit drugs.

[Current prevalence of Clonorchis sinensis infection in endemic areas of China].

OBJECTIVE: To investigate the current prevalence of Clonorchis sinensis infection in the known endemic areas of China. METHODS: According to different endemic level and river system, the lamination stochastic group sampling was carried out in 27 Provinces/Municipalities/Autonomous Regions (P/M/A) from 2002 to 2004. Eggs in stool were examined by modified Kato-Katz thick smears method. RESULT: A total of 5,230 cases of Clonorchis infection was found from 217,829 sampled population of 27 P/M/A, and the mean prevalence was 2.40% by which the number of infected people was estimated to be 12.49 millions. The geometry mean worm egg density was 2,208 EPG (stool). Light, moderate and heavy infection occupied 78.93%, 17.40%, and 3.67% respectively. Infection was found in 19 out of 27 P/M/A investigated, with the highest prevalence in the provinces of Guangdong (16.42%, 2,278/13,876), Guangxi (9.75%, 1,365/13,990) and Heilongjiang (4.72%, 636/13,458). Males showed higher prevalence (2.94%, 3,267/111,262) than females (1.84%, 1,963/106,567). There was infection in all age groups,with the highest prevalence in the group of 50 to 59 years (9.16%). By occupations, more infections were found in businessmen (13.42%, 124/924), followed by factory workers ((7.9%, 298/3,773), and retirees and veterans (5.28%, 70/1,327). Adults showed heavier infection in some provinces where the main infection way was raw fish-eating, but in children in other provinces. For geographical distribution, the terrain in network of rivers and lakes showed higher prevalence (5.23%, 687/13,125) than the hilly terrain (2.34%, 1,111/47,552). Prevalence showed family-aggregation feature. CONCLUSION: Clonorchiasis is widely endemic in China with a high prevalence in some provinces. Male adults have been more involved with a family aggregation.

[Follow-up survey on the imported cases of lymphatic filariasis in Guangdong Province].

OBJECTIVE: To make a survey on the possible imported cases with microfilaremia from a recently identified outbreak focus in Fuchuan County of Guangxi Region. METHODS: Moving people as temporary laborers from Guangxi and periphery residents were screened by thick blood smears for microfilariae and ELISA for specific IgG4 in Sept 2007. RESULTS: 989 persons in cities/districts of Nansha, Panyu, Shantou and Zhongshan were investigated. Among 150 persons from Changtang administrative village of Guangxi, 47 were from two outbreak focuses, Ganshang and Yinshan natural villages in Changtang. Three cases with microfilaremia were found, with a microfilaraemia density of 24, 20 and 2 per 120 microl blood, respectively. The 3 cases were all from the 2 natural villages, with a microfilaraemia positive rate of 6.4% (3/47). Meanwhile, anti-filaria IgG4 was detected for other 1 501 persons, including those from the outbreak focuses, from surr-founding areas and from non-endemic area as control, fifteen cases were found with IgG4 positive. Of the 15 positive cases, 12 were from the 2 outbreak natural villages, with a positive rate of 25.5%, higher than those of others (chi2=295.83, P < 0.01). The cases with IgG4 positive but microfilaremia negative were reexamined by ICT kit and all showed negative. CONCLUSION: It is suggested that no further transmission occurs although microfilaremia cases have been imported from the outbreak focus of Guangxi.

Small-scale spatial analysis of intermediate and definitive hosts of Angiostrongylus cantonensis.

BACKGROUND: Angiostrongyliasis is a food-borne parasitic zoonosis. Human infection is caused by infection with the third-stage larvae of Angiostrongylus cantonensis. The life cycle of A. cantonensis involves rodents as definitive hosts and molluscs as intermediate hosts. This study aims to investigate on the infection status and characteristics of spatial distribution of these hosts, which are key components in the strategy for the prevention and control of angiostrongyliasis. METHODS: Three villages from Nanao Island, Guangdong Province, China, were chosen as study area by stratified random sampling. The density and natural infection of Pomacea canaliculata and various rat species were surveyed every three months from December 2015 to September 2016, with spatial correlations of the positive P. canaliculata and the infection rates analysed by ArcGIS, scan statistics, ordinary least squares (OLS) and geographically weighted regression (GWR) models. RESULTS: A total of 2192 P. canaliculata specimens were collected from the field, of which 1190 were randomly chosen to be examined for third-stage larvae of A. cantonensis. Seventy-two Angiostrongylus-infected snails were found, which represents a larval infection rate of 6.1% (72/1190). In total, 110 rats including 85 Rattus norvegicus, 10 R. flavipectus, one R. losea and 14 Suncus murinus were captured, and 32 individuals were positive (for adult worms), representing an infection rate of 29.1% of the definitive hosts (32/110). Worms were only found in R. norvegicus and R. flavipectus, representing a prevalence of 36.5% (31/85) and 10% (1/10), respectively in these species, but none in R. losea and S. murinus, despite testing as many as 32 of the latter species. Statistically, spatial correlation and spatial clusters in the spatial distribution of positive P. canaliculata and positive rats existed. Most of the spatial variability of the host infection rates came from spatial autocorrelation. Nine spatial clusters with respect to positive P. canaliculata were identified, but only two correlated to infection rates. The results show that corrected Akaike information criterion, R2, R2 adjusted and σ2 in the GWR model were superior to those in the OLS model. CONCLUSIONS: P. canaliculata and rats were widely distributed in Nanao Island and positive infection has also been found in the hosts, demonstrating that there was a risk of angiostrongyliasis in this region of China. The distribution of positive P. canaliculata and rats exhibited spatial correlation, and the GWR model had advantage over the OLS model in the spatial analysis of hosts of A. cantonensis.