[Identification of Cervi Cornu (Cervus elaphus) and its formula granules based on PCR-RFLP].

Cervi Cornu is the ossified antler, or the base antler that falls off in the spring of the following year after the pilose antler is sawn off from Cervus elaphus or C. nippon, as a precious traditional Chinese medicine, has been recognized for its medicinal value and widely used in clinical practice. However, the origins of Cervi Cornu are miscellaneous, and Cervi Cornu is even mixed with adulterants in the market. Currently, there is a shortage of ways to identify Cervi Cornu and no standard to control the quality of Cervi Cornu. So it is valuable to develop a way to effectively identify Cervi Cornu from the adulterants. In this study, the differences in the mitochondrial barcode cytochrome b(Cytb) gene sequences of C. elaphus, C. nippon and their related species were compared and the specific single nucleotide polymorphism(SNP) sites on the Cytb sequences of Cervi Cornu were screened out. According to the screened SNPs, Cervi Cornu-specific primers dishmy-F and dishmy-R were designed. The PCR system was established and optimized, and the tolerance and feasibility of Taq polymerases and PCR systems affecting the repeatability of the PCR method were investigated. The amplification products of C. elaphus and C. nippon were digested using the restriction enzyme MseⅠ. The results showed that after electrophoresis of the product from PCR with the annealing temperature of 56 ℃ and 35 cycles, a single specific band at about 100 bp was observed for C. elaphus samples, and the product of C. elaphus samples was 60 bp shorter than that of C. nippon samples. There was no band for adulterants from other similar species such as Alces alces, Rangifer tarandus, Odocoileus virginianus, O. hemionus, Cap-reolus pygargus, Przewalskium albirostis and negative controls. The polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method established in this study can quickly and accurately identify Cervi Cornu originated from C. elaphus in crude drugs, standard decoctions, and formula granules, and distinguish the origins of Cervi Cornu products, i.e., C. nippon and similar species. This study can be a reference for other studies on the quality standard of other formula granules of traditional Chinese medicines.

Integrated 4D label-free proteomics and data mining to elucidate the effects of thermal processing on crisp grass carp protein profiles.

The crisp grass carp (CGC; Ctenopharyngodon idellus C. et V.), known for its unique texture and flavour, is a culinary delicacy whose quality is significantly influenced by thermal processing. This study employed 4D label-free proteomics and data mining techniques to investigate the proteomic changes in CGC muscle tissue induced by various heating temperatures. CGC samples were subjected to a series of heat treatments at increasing temperatures from 20 °C to 90 °C. Proteins were extracted, digested, and analysed using high-resolution mass spectrometry. The proteomic data were then subjected to extensive bioinformatics analysis, including GO and KEGG pathway enrichment. We identified a total of 1085 proteins, 516 of which were shared across all the temperature treatments, indicating a core proteome responsible for CGC textural properties. Differential expression analysis revealed temperature-dependent changes, with significant alterations observed at 90 °C, suggesting denaturation or aggregation of proteins at higher temperatures. Functional enrichment analysis indicated that proteins involved in amino acid metabolism, glutathione metabolism, and nucleotide metabolism were particularly affected by heat. Textural analysis correlated these proteomic changes with alterations in CGC quality attributes, pinpointing 70 °C as the optimum temperature for maintaining the desired texture. A strong positive correlation between specific upregulated proteins was identified, such as the tubulin alpha chain and collagen alpha-1(IV) chain, and the improved textural properties of CGC during thermal processing, suggesting their potential as the potential biomarkers. This study offers a comprehensive proteomic view of the thermal stability and functionality of CGC proteins, delivering invaluable insights for both the culinary processing and scientific management of CGC. Our findings not only deepen the understanding of the molecular mechanisms underpinning the textural alterations in CGC during thermal processing but also furnish practical insights for the aquaculture industry. These insights could be leveraged to optimize cooking techniques, thereby enhancing the quality and consumer appeal of CGC products.

[Irritant toxicity and lectin content of different processed products of Pinelliae Rhizoma].

The present study aims to investigate the correlation between irritant toxicity variation and lectin content variation during the processing of Pinelliae Rhizoma products and to explore the feasibility of Western blot as a method for the detection of lectin. We processed Pinelliae Rhizoma Praeparatum Cum Alumine, Pinelliae Rhizoma Praeparatum, and Pinelliae Rhizoma Praeparatumcum Zingibere et Alumine to different degrees and then analyzed their irritant toxicity via Draize rabbit eye test. Western blot was employed to determine the lectin content in Pinelliae Rhizoma products processed with different methods. The correlation between toxicity variation and lectin content variation was then analyzed. Different decoction pieces of Pinelliae Rhizoma were collected for the determination of lectin content. The three processed products of Pinelliae Rhizoma showed gradually decreased toxicity and lectin content as the processing continued. The decreasing trend of lectin content was consistent with that of irritant toxicity during processing, which indicated that the change in lectin content could reflect the trend of irritant toxicity. No band of lectin appeared in the Western blot of processed products of Pinelliae Rhizoma, which suggested that western blotting can be used for the detection of toxic lectin in the processed products of Pinelliae Rhizoma. Lectin should not be detected in the Pinelliae Rhizoma products processed according to the methods in the Chinese Pharmacopoeia.

[Identification of specific peptides from Bubali Cornu by ultra-performance liquid chromatography-tandem mass spectrometry].

This paper explored the specific peptides from Bubali Cornu by ultra-performance liquid chromatography-tandem mass spectrometry and based on mathematics set theory. Following the profile analysis of peptides from Bubali Cornu, Bovis Grunniens Cornu, Caprae Hircus Cornu, and Suis Cornu by nano LC-LTQ-Obitrap-MS after digestion with trypsin, the relationship of peptide composition among different samples was analyzed using the mathematics set theory. The ones that existed only in the Bubali Cornu set rather than in any other set were considered as the specific peptides of Bubali Cornu. The further bioinformatic analysis revealed four specific peptides from Bubali Cornu, whose specificity was verified by UPLC-QQQ-MS. The results showed that these four peptides could be used for distinguishing Bubali Cornu from Caprae Hircus Cornu and Suis Cornu. This study has provided a rapid and simple method for seeking the specific peptides in animal medicines, which can be utilized for quality evaluation of animal medicines, thus making them authenticable and traceable.

[Effect of different parts of Pinelliae Rhizoma Decoction against airway inflammation and analysis of effective components].

This study investigated the material basis and mechanism of Pinelliae Rhizoma Decoction in the treatment of airway inflammation. The cigarette smoke combined with lipopolysaccharide(LPS) was used to induce an airway inflammation model in mice. The expression levels of IL-6 and IL-8 in the bronchoalveolar lavage fluid(BALF) and the phosphorylation levels of p38 and IκB in the lungs of mice were taken as indexes to screen the effective extracts by system solvent extraction from Pinelliae Rhizoma Decoction(dichloromethane extract, ethyl acetate extract, n-butanol extract, etc.). Meanwhile, the human bronchial epithelial(16-HBE) cell model of cigarette smoke extract(CSE)-induced injury was established, and the mRNA expression levels of IL-6 and IL-8 and the phosphorylation levels of p38 and IκB proteins were also taken as indexes to evaluate the anti-inflammatory effect of different extracts of Pinelliae Rhizoma Decoction. The results showed that Pinelliae Rhizoma Decoction significantly antagonized airway inflammation in mice by down-regulating the expression levels of IL-6 and IL-8 in mice with airway inflammation and 16-HBE cells with CSE-induced injury and inhibiting the phosphorylation levels of p38 and IκB. The dichloromethane and ethyl acetate extracts of Pinelliae Rhizoma Decoction showed significant anti-inflammatory effects, while such effects of other extracts were not prominent. Furthermore, the database of Pinelliae Rhizoma composition was constructed, and the components in effective extracts were analyzed by HPLC-TOF-MS and Nano-LC-MS/MS. As revealed by the results, the compositions of the two effective extracts were similar with 36 common components. They were combined and then divided into Pinelliae Rhizoma alkaloids(PTAs) and Pinelliae Rhizoma non-alkaloids(PTNAs) by 732 cation-exchange resin. Further in vitro investigation confirmed the significant anti-inflammatory effect of PTNAs, while such effect of PTAs was not manifest. The MS analysis showed 172 peptides and 7 organic acids in PTNAs. The peptide content in PTNAs was 63.5% measured by quantitative analysis of BCA assay, and the organic acid content was 9.92% by potentiometric titration method. The findings of this study suggested that Pinelliae Rhizoma Decoction could antagonize airway inflammation in mice by inhibiting phosphorylation of p38 and IκB and blocking the activation of MAPK and NF-κB signaling pathways, and the effective components were related to the peptides and organic acids in PTNAs. The above results lay a foundation for the research on the mechanism and material basis of Pinelliae Rhizoma in antagonizing airway inflammation.

Integrating chemical similarity and bioequivalence: A pilot study on quality consistency evaluation of dispensing granule and traditional decoction of Scutellariae Radix by a totality-of-the-evidence approach.

There is an increasing focus on the quality consistency evaluation of dispensing granule in traditional Chinese medicines (TCMs). According to the guideline from Chinese Pharmacopoeia Commission, the substantial equivalence of dispensing granule and traditional decoction should be determined, and the chromatographic fingerprint has been recommended as a comprehensive qualitative approach to assess the quality consistency between dispensing granule and traditional decoction. However, a high-degree chemical similarity does not equal a bioequivalence. Attempting to realize the quality evaluation by integrating chemical consistency and bioequivalence, we herein proposed a totality-of-the-evidence approach based on clustering analysis and equivalence evaluation taking the dispensing granule and traditional decoction of Scutellariae Radix (SR) as a typical case. Chemical fingerprints were developed by high performance liquid chromatography coupled with photodiode array detector and quadrupole time-of-flight mass spectrometry (HPLC-PDA/QTOF-MS). Subsequently, a feature selection strategy, integrated linear and nonlinear correlation analysis, was carried out to assess the correlation between chemical profiles and biological activities. Finally, quality consistency between the dispensing granule and the traditional decoction was determined by bioactive marker-guided hierarchical clustering analysis (HCA), k-means clustering method and bioequivalence evaluation. The available evidence suggested that not all the dispensing granule of SR were sufficiently similar to the traditional decoction. This study provides an applicable methodology for quality consistency evaluation of dispensing granule and traditional decoction in TCMs.

Activities of Endogenous Lipase and Lipolysis Oxidation of Low-Salt Lactic Acid-Fermented Fish (Decapterus maruadsi).

There is increasing demand for low-salt meat products that retain traditional flavors. In this study, dry-salted fish (Decapterus maruadsi) were processed by 2 methods to obtain traditional salted fish (HS) and low-salt lactic acid-fermented fish (LAF). The relationship between lipolysis and lipid oxidation was investigated by evaluating changes in endogenous lipase (lipolytic enzymes; lipoxygenase, LOX), free fatty acid composition, thiobarbituric acid reactive substances (TBARS), and peroxide value (POV) during processing. Lipolytic enzyme activity showed a decreasing trend, in general. LOX activity initially increased and eventually decreased. Phospholipase, acid lipase, and neutral lipase activity was 0.33, 0.17, 0.57 (in HS) and 0.39, 0.25, 0.67 (in LAF) times in the final product than the activity levels observed in fresh fish. A principal component analysis indicated that phospholipase and neutral lipase play major roles in promoting lipid hydrolysis (in HS and LAF), the correlation between lipolytic activity and lipid oxidation in HS is greater than the correlation in LAF, and the contribution of LOX to lipid oxidation was minor in salted fish.

[Analysis of schistosomiasis endemic situation in Yizhou City from 2011 to 2014].

OBJECTIVE: To evaluate the effect of schistosomiasis control in Yizhou City, Guangxi Zhuang Autonomous Region, from 2011 to 2014, so as to provide the evidence for formulating the further prevention and control strategy. METHODS: The schistosomiasis surveillance data were collected and analyzed in Yizhou City from 2011 to 2014. RESULTS: From 2011 to 2014, 770.38 hm² was surveyed for the Oncomelania hupensis snails, and two snail infested sites were found, while no infected snails were found. Totally 3,524 residents were tested by ELISA for Schistosoma japonicum infection, and 38 cases were positive. The positive rate of 2013 was significantly higher than those of other three years (χ² = 15.08, P < 0.05). Totally 432 rats and 28 dogs were dissected and 1,697 cattle were examined by the stool test, but no positive cases were found. CONCLUSIONS: The indicators of schistosomiasis surveillance are basically stable in Yizhou City.

A new mathematical equation for the evaluation of the compression behavior of pharmaceutical materials.

A new mathematical equation characterizing the compression of pharmaceutical materials is presented. This equation presumed that the rate of change of the compressible volume of powder with respect to the pressure is proportional to the compressible volume. The new model provided a good fit to several model substances employing non-linear regression techniques. The validity of the model had been verified with experimental results of various pharmaceutical powders according to the Akaikes informatics criterion (AIC) and the sum of squared deviations (SS). The parameter of the new model might reflect quantitatively the fundamental compression behaviors of the powders. It had demonstrated that the proposed model could well predict the compaction characteristics of solid particles like the Kawakita model.

[Evaluation with compression equations of compression behavior of pellets with different intragranular pore volumes].

Microcrystalline cellulose (MCC), calcium phosphate (DCP)/MCC (4:1, w/w) and lactose (Lac)/MCC (4:1) pellets with different intragranular porosity were prepared in an extrusion-spheronizator and three volume ratios of ethanol/water were used as binder agents to prepare pellets. The compression behaviors of these pellets with different intragranular pore volume were evaluated with the parameters of Kawakita model. The results showed that high pore volume of pellets made up of MCC had the best compressibility and low pore volume of pellets had a poor compactibility. However, the compressibility of different porosity of pellets made up of DCP/MCC (4:1) or Lac/MCC (4:1) was good, but they were not significantly different. The reason might be the main compression mechanism of high porosity of MCC pellets was plastic deformation and that of DCP/MCC pellets or Lac/MCC pellets was not plastic deformation but fragmentation. These results can be observed directly by the SEM photographs. According to these results, the conclusion could be drawn that high porosity MCC pellets and different porosity DCP/MCC pellets and Lac/MCC pellets can be used as cushion granules to maintain the original shape and release characteristics of drug pellets when pellets were tabletted.

[Preparation of tablets containing enteric-coated diclofenac sodium pellets].

Fluidized-bed manufactured enteric-coated diclofenac sodium pellets were compressed into tablets. The blend of two aqueous acrylic resins dispersion in different ratios, Eudragit NE30D and Eudragit L30D-55, were used to prepare enteric-coated diclofenac sodium pellets of different particle sizes and coating level. The cushioning pellets with different properties and these enteric-coated pellets were compressed into tablets in different proportions. The drug release of the tablets containing these pellets would be lower than 10% in 2 h in simulated gastric fluid, but reach (83 +/- 2.42)% in 1 h in simulated enteric fluid. The mixture of Eudragit NE30D and Eudragit L30D-55 could be used to prepare enteric pellets which are suitable for compression. The cushioning pellets which were composed of stearic acid/microcrystalline cellulose (4:1, w/w) could avoid rupture of the coating of pellets during the compression.

[Study on evaluation index of acupoint transdermal administration--drug delivery coefficient].

OBJECTIVE: To investigate the correlation of skin electric resistance changes with the blood drug content in acupoint transdermal administration, and to establish an new evaluation index for drug delivery efficiency through acupoints. METHODS: Twenty-four rabbits were randomly divided into an observation group and a control group. In the observation group, aminophylline was administrated through "Feishu" (BL 13), "Geshu" (BL 17) and "Danzhong" (CV 17) which are commonly used for treatment of bronchial asthma, and the control group through sham-points on the back. The skin resistance and plasma aminophylline content were determined after application of aminophylline to the points, and their changes with time were observed. The ratio of Css/Rss at stability was defined as delivery coefficient (DC) which reflects the efficiency of delivering drug at acupoints and sham acupoints. RESULTS: Both the plasma aminophylline and the skin resistance value tended to steady about 6-8 h after application of aminophyiophylline. The DC in the acupoints was higher than that in the sham-acupoints (P < 0.01). And there was significant difference as DC of the "Feishu"was significant difference (BL 13) and "Danzhongas DC of the "Feish" (BLCV 17) compared with that of "Geshu" (BL 17) (P < 0.01). CONCLUSION: The bigger the DC is, the higher the efficiency of drug delivery is; the efficiency of drug delivery through acupoints is higher than that through sham-acupoints.

[The release behavior of the combined system of diltiazem hydrochloride delayed-onset sustained-release pellets and modeling by mathematics method].

AIM: To prepare the combined system of diltiazem hydrochloride delayed-onset sustained-release pellets in order to make time-specific drug delivery system. The drug can release from the system sustained after a predetermined lag time, and the release behavior can continue till 24 hour after administrating the formulation. According to the concept of chronotherapy, the combined system is useful to improve the pharmacotherapy of cardiovascular diseases. METHODS: The velocity-time curve of the drug release from the multiple-unit system containing pellets was consistent with the fluctuation curve following time of blood pressure and heart ratio. So the velocity-time curve was selected to describe the release behavior of the combined system. The velocity-time equation describing the release behavior of two kinds of pellets was deduced by non-linear least square model fit. And zero-order kinetics equation was adopted to fit the release behavior of different combinations which were composed of different proportion of two kinds of pellets. The velocity-time equation describing the release behavior of the combinations was deduced by non-linear least square model fit, too. The difference of combinations in velocity-time curves between theoretical value and test value was compared. RESULTS: The results showed that the test values were closely approximate to the theoretical values. Therefore, the multiple unit drug delivery system can be described by adding the velocity-time equations of different pellets to calculate the theoretical equations. CONCLUSION: A multiple-unit combined system containing different coated pellets, as a novel delayed-onset sustained-release system, was prepared. Then a time-specific drug delivery system has been made. The programmed drug delivery system could be predicted by adding the velocity-time equation of each kind of pellets to calculate the theoretical equations. characterized by mathematics equation. The release behavior of pellets system could be characterized by mathematics equation.

[Studies on combined release behaviors of versatile mini-tablets in capsule systems and fittings of their mathematical model].

AIM: To prepare nifedipine (NP) rapid release mini-tablet, sustained release mini-tablets, pulsed release mini-tablets and delayed-onset sustained release mini-tablets and develop multiplied pulsed drug delivery system (DDS), site-specific DDS, zero-order DDS and quick/slow DDS by various ways. METHODS: Velocity-time (v-t) equation of each mini-tablet was deduced by non-linear least square model fit. The difference of combinations in v-t profiles between theoretical value and test value was compared. RESULTS: According to the v-t equations, the combined release behaviors were observed directly from v-t profiles and the test values coincided with the theoretical profiles. CONCLUSION: The programmed DDS, which consist of a variety of mini-tablets with different dosages and combinations in capsules, could be predicted by summing up the v-t equation of each tablet.