Epidemic situation and diagnosis of imported malaria before and after malaria elimination in Nanjing City, Jiangsu Province / 中国血吸虫病防治杂志

Objective To analyze the epidemiological characteristics and diagnosis of imported malaria before and after malaria elimination in Nanjing City of Jiangsu Province, so as to provide the scientific evidence for formulating the malaria control strategy after malaria elimination. Methods Data pertaining to the epidemic situation and individual investigation of malaria in Nanjing City before (from 2012 to 2016) and after malaria elimination (from 2017 to 2020) were captured from the National Notifiable Communicable Disease Reporting System and the Information System for Parasitic Diseases Control and Prevention and were analyzed statistically. Results A total of 178 malaria cases were reported in Nanjing City from 2012 to 2020, and all were imported cases. There were 99 malaria cases reported before malaria elimination in Nanjing City, including 78 cases with Plasmodium falciparum malaria (78.79%), 5 cases with P. vivax malaria (5.05%), 10 cases with P. ovale malaria (10.10%), 3 cases with P. malariae malaria (3.03%) and 3 cases with mixed infections (3.03%), and 79 malaria cases reported after elimination, including 63 cases with P. falciparum malaria (79.75%), 5 cases with P. vivax malaria (6.33%), 9 cases with P. ovale malaria (11.39%), 2 cases with P. malariae malaria (2.53%). There was no significant difference in the proportion of each type of malaria cases in Nanjing City before and after malaria elimination (χ2 =2.400, P > 0.05). Malaria cases mainly acquired Plasmodium infections in African regions, and no significant difference was seen in the proportion of malaria cases returning to Nanjing City from African countries before and after malaria elimination (χ2 = 0.093, P > 0.05). The number of malaria cases peaked in Nanjing City in January and during the period from May to July before elimination, and there was no apparent seasonal variation in the distribution of malaria cases after elimination. The proportion of malaria cases living in Nanjing City was significantly greater after malaria elimination than before elimination (72.15% vs. 55.56%; χ2 = 5.187, P = 0.023). The proportions of businessmen and international students were both 5.05% before malaria elimination, and increased to 15.19% and 13.92% after elimination, respectively (χ2 = 5.229 and 4.229, both P values < 0.05). The percentage of definitive diagnosis of malaria at initial diagnosis was 18.75% in county-level hospitals before malaria elimination and increased to 61.11% after elimination (χ2 = 6.275, P = 0.012), while the proportion of malaria cases with definitive diagnoses in county-level hospitals was 4.04% before malaria elimination and increased to 13.92% after elimination (χ2 = 5.562, P = 0.018). During the period from 2012 to 2020, the proportion of malaria cases with definitive diagnoses within 1 to 3 days post-admission increased from 27.27% in Nanjing City before malaria elimination to 45.57% after elimination (χ2 = 6.433, P = 0.011). Conclusions The epidemic situation of imported malaria remains serious in Nanjing City during the post-elimination stage, and malaria parasite infections predominantly occur in African regions. In addition, there are changes in regional and occupational distributions of malaria cases and the diagnostic capability of malaria increases in county-level hospitals in Nanjing City after malaria elimination. Further improvements in the malaria surveillance system and the diagnostic and treatment capability of malaria in medical institutions at each level are required to consolidate malaria elimination achievements in Nanjing City.

Antithrombotic effects of ethyl acetate fraction of Curcuma kwangsiensis / 中成药

AIM To study the antithrombotic effects and mechanism of ethyl acetate fraction of Curcuma kwangsiensis S.G.Lee et C.F.Liang (CKEAF).METHODS To observe the antithrombotic effects of CKEAF on tail thrombotic mouse models and arteriovenous bypass thrombotic rat models induced by carrageenan,and in vivo thrombosis rat models induced by FeCl3.The serum levels of ET-1,6-keto-PGF1α,TXB2 in rats were measured by enzyme linked immunosorbent assay (ELISA),and the level of NO was determined by nitrate reductase method.Rat models of acute blood stasis were further established for hemorheological study.RESULTS CKEAF significantly decreased the number and length of blackened thrombotic tail of carrageenan-treated mice.Rat models shared significant wet weight loss in arteriovenous bypass thrombosis and in vivo thrombosis,increased levels of NO,6-keto-PGF1α,decreased levels of ET-1,TXB2,and decreased whole blood and plasma viscosity (rats of acute blood stasis model).CONCLUSION Significant antithrombotic effects by CKEAF may contribute to elevated levels of NO,6-keto-PGF1α,and decreased levels of ET-1,TXB2,and lowered whole blood and plasma viscosity.

Pharmacokinetics of the prototype and hydrolyzed carboxylic forms of ginkgolides A, B, and K administered as a ginkgo diterpene lactones meglumine injection in beagle dogs / 中国天然药物

Ginkgo diterpene lactones meglumine injection (GDLI) is a commercially available product used for neuroprotection. However, the pharmacokinetic properties of the prototypes and hydrolyzed carboxylic forms of the primary components in GDLI, i.e., ginkgolide A (GA), ginkgolide B (GB), and ginkgolide K (GK), have never been fully evaluated in beagle dogs. In this work, a simple, sensitive, and reliable method based on ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) was developed, and the prototypes and total amounts of GA, GB, and GK were determined in beagle dog plasma. The plasma concentrations of the hydrolyzed carboxylic forms were calculated by subtracting the prototype concentrations from the total lactone concentrations. For the first time, the pharmacokinetics of GA, GB, and GK were fully assessed in three forms, i.e., the prototypes, the hydrolyzed carboxylic forms, and the total amounts, after intravenous administration of GDLI in beagle dogs. It was shown that ginkgolides primarily existed in the hydrolyzed form in plasma, and the ratio of hydrolysates to prototype forms of GA and GB decreased gradually to a homeostatic ratio. All of the three forms of the three ginkgolides showed linear exposure of AUC to the dosages. GA, GB, and GK showed a constant half-life approximately 2.7, 3.4, and 1.2 h, respectively, which were consistent for the forms at three dose levels (0.3, 1.0, and 3.0 mg·kg) and after a consecutive injection of GDLI for 7 days (1.0 mg·kg).

Impact of parameter fluctuations on the performance of ethanol precipitation in production of Re Du Ning Injections, based on HPLC fingerprints and principal component analysis / 中国天然药物

The present study was designed to determine the relationships between the performance of ethanol precipitation and seven process parameters in the ethanol precipitation process of Re Du Ning Injections, including concentrate density, concentrate temperature, ethanol content, flow rate and stir rate in the addition of ethanol, precipitation time, and precipitation temperature. Under the experimental and simulated production conditions, a series of precipitated resultants were prepared by changing these variables one by one, and then examined by HPLC fingerprint analyses. Different from the traditional evaluation model based on single or a few constituents, the fingerprint data of every parameter fluctuation test was processed with Principal Component Analysis (PCA) to comprehensively assess the performance of ethanol precipitation. Our results showed that concentrate density, ethanol content, and precipitation time were the most important parameters that influence the recovery of active compounds in precipitation resultants. The present study would provide some reference for pharmaceutical scientists engaged in research on pharmaceutical process optimization and help pharmaceutical enterprises adapt a scientific and reasonable cost-effective approach to ensure the batch-to-batch quality consistency of the final products.

Metabolism of six saponins by rat intestinal bacteria in vitro / 中国中药杂志

To investigate the metabolism of six saponins by rat intestinal bacteria in vitro.Six saponins, including notoginsenoside R₁, ginsenoside Rg₁, ginsenoside Rg₂, ginsenoside Re, ginsenoside Rd and ginsenoside Rb₁, were incubated for 8 and 24 h with rat intestinal bacteria under anaerobic environment, respectively. After the samples were precipitated by acetonitrile and extracted with ethyl acetate, LC-Q-TOF-MS/MS was applied for the qualitative analysis of the metabolites. The potential metabolites in rat feces were analyzed by comparing the total ion current of the test samples and blank samples and analyzing the quasi-molecular ion and fragment ion of all chromatograms. The results showed that six saponins could be easily metabolized by rat intestinal bacteria. Notoginsenoside R₁ was mainly metabolized into five metabolites, and it's metabolic pathway was notoginsenoside R₁→ginsenoside Rg₁→ginsenoside Rh₁ and ginsenoside F₁→protopanaxatriol→dehydrogenated protopanaxatriol. Ginsenoside Rg₁ was mainly metabolized into four metabolites, and it's metabolic pathway was ginsenoside Rg₁→ginsenoside Rh₁ and ginsenoside F₁→protopanaxatriol→dehydrogenated protopanaxatriol. Ginsenoside Rg₂ was mainly metabolized into two metabolites, and it's metabolic pathway was ginsenoside Rg₂→ protopanaxatriol→dehydrogenated protopanaxatriol. Ginsenoside Re was mainly metabolized into four metabolites, and it's metabolic pathway was ginsenoside Re→ginsenoside Rg₂→ginsenoside F₁→protopanaxatriol→dehydrogenated protopanaxatriol. Ginsenoside Rd was mainly metabolized into four metabolites, and it's metabolic pathway was ginsenoside Rd→ginsenoside Rg₃ and ginsenoside F₂→ginsenoside Rh₂→protopanaxadiol. Ginsenoside Rb1 was mainly metabolized into five metabolites, and it's metabolic pathway was ginsenoside Rb₁→ginsenoside Rd→ginsenoside Rg₃ and ginsenoside F₂→ginsenoside Rh₂→protopanaxadiol. In summary, six saponins could be quickly metabolized by rat intestinal bacteria in vitro. Their major metabolic pathways were deglycosylation and dehydrogenation.

Study on limit detection of flavones in diterpene ginkgolides meglumine injection materials by LC-MS and HPLC-DAD / 中国中药杂志

Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.

Determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials by LC-MS/MS / 中国中药杂志

To develop a LC-MS/MS method for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials, the column was Agilent ZORBAX Eclipse plus C18 (3.0 mm x 50 mm, 1.8 µm), and the mobile phase consisted of methanol-water (containing 0.2% formic acid) (95:5) at a flow rate of 0.5 mL · min(-1). The multiple reaction ion monitoring (MRM) with an ESI interface in the negative ion mode was selected. The results showed that the linear ranges of five kinds of ginkgolic acids were in the range of 0.2-36.0 µg · L(-1) (r ≥ 0.999 5). The lowest limit of quantification (LOQ) of ginkgo acid C13: 0, C15:1, C17:2, C15:0 and C17:1 were 0.18, 0.18, 0.21, 0.10 and 0.20 µg · L(-1), respectively. The average recovery was between 73.28% and 87.56%, and the average content of total ginkgolic acids in three batches of samples was in the range of 0.023-0.028 µg · g(-1), which was much lower than 2 µg · g(-1) prescribed in drug registration standards. This method is simple and rapid with high sensitivity, which can be used for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials.

Similarity between leaves of Nauclea officinalis and stems of Nauclea officinalis / 中国中药杂志

The study is to develop a method to determine 3 batches leaves of Nauclea officinalis and stems of N. officinalis by HPLC. The differences between strictosamide contents and fingerprints was compared, then chromatographic peak of fingerprints was validated with the assistance of LC-MS. The strictosamide contents in stems of N. officinalis were higher than leaves of N. officinalis. The main chemical composition in leaves of N. officinalis and stems of N. officinalis were alkaloid which revealed by LC-MS. There are 7 chemical compositions were same between them, but the chemical composition in leaves of N. officinalis is more than stems of N. officinalis. This provides a scientific basis for the development of the potential medicinal value of leaves of N. officinalis and the sustainable utilization of N. officinalis.

Quantitative and qualitative evaluation on tablets of Ginkgo biloba leaves using fingerprint and LC-MS analysis / 中国中药杂志

A reasonable method for the quality control of tablets of Ginkgo biloba leaves was established in this paper. The total flavonol glycosides and terpene lactones of G. biloba tablets were quantified by HPLC. Totally, 16 batches of the commercially available tablets of G. biloba leaves were determined. Among of them, 2 batches were unqualified in the content of total flavonol glycosides, and 3 batches were unqualified in the content of terpene lactones. A validated HPLC fingerprint method was established to evaluate the commercially available tablets of G. biloba leaves with the assistance of LC-MS. Sixteen batches showed the similarity of 0.763-0.989. There were 31 fingerprint chromatogram peaks were identified as flavonoids compositions by LC-MS. This provides a research idea for the quality control of tablets of G. biloba leaves.

Rapid identification of six chemical constituents in Guizhi Fuling capsule by DART-Q-TOF-MS / 中国中药杂志

In order to establish a rapid method for identifying six constituents in Guizhi Fuling capsule, Q-TOF with DART ion source was used to perform the direct analysis of compounds in Guizhi Fuling capsule. The DART sampler delivery rate was 0.2 mm s(-1). The temperature of helium gas of DART was 450 degrees C. The capillary voltage was kept at 1 000 V. The temperature of the drying gas of Agilent 6538 Q-TOF MS was set at 350 degrees C. The flow rate of the drying gas of MS was set at 3.5 L x min(-1). The MS scan range was m/z 50-1 000. Based on accurate mass measurements and the elemental compositions of the product ions and fragmentation patterns of reference conpounds, six components, amygdalin, paeonol, paeoniflorin, cinnamic acids, gallic acid, benzoic acid were identified rapidly. The method can rapidly identify six chemical constituents in three batch of Guizhi Fuling capsule. The DART-Q-TOF-MS method is simple, rapid and specific and it can be used for rapid identification and characterization of compounds in traditional Chinese medicines.

Quality evaluation of guizhi fuling capsule using self-control method of reference Chinese medicine preparation / 中国中药杂志

Taking guizhi fuling capsule (GZFL) for instance, a new method about reference Chinese medicine preparation which was used as standard substance for the quality evaluation of complex Chinese medicine preparation by the fingerprint of reference preparation instead of standard fingerprint was proposed. It could eliminate the errors from different instruments, chromatographic columns and solve the problem of similarity matching in the absence of standard fingerprint. The qualification of reference GZFL was evaluated according to the quality control method of GZFL from Chinese Pharmacopoeia. Then multiple batches of GZFL were estimated, taking fingerprint of reference preparation and standard fingerprint as references, respectively, at different instruments and chromatographic columns. Finally, the packaging and expiration date for reference GZFL were confirmed according to the results of stability investigation. The results indicated that the fingerprint of reference GZFL could be used to assess the quality of GZFL better than standard fingerprint. The data of accelerated stability and long-term stability test demonstrated that reference GZFL was stable in the conditions of double blister package. Therefore, reference GZFL can be used as standard substance in quality control of GZFL.