Sleep disorders are increasingly prevalent within the population, and optimal sleep is essential for overall health. The sleep-wake cycle is a multifaceted process influenced by a variety of factors, including several neurotransmitters such as acetylcholine, norepinephrine, serotonin, histamine, dopamine, orexin, and gamma-aminobutyric acid (GABA). The activity of these neurotransmitters is further modulated by numerous nutrients involved in their metabolic pathways. In recent years, GABA has garnered significant attention due to its critical role in sleep regulation. This review aims to examine the mechanisms through which GABA affects sleep, its clinical implications, as well as recent advancements in research and future directions in this field.

OBJECTIVE To investigate the biosynthetic pathway of C₂₁ steroidal compounds in Cynanchum otophyllum Schneid. METHODS Metabolomics and transcriptomics were used to compare and analyze the relative contents of C₂₁ steroids in the roots, stems and leaves of C. otophyllum. Then, the genes related to C₂₁ steroid biosynthesis in C. otophyllum were screened through Kyoto Encyclopedia of Genes and Genomes(KEGG) database annotation. Finally, some differentially expressed genes (DEGs) were verified by quantitative real-time PCR. RESULTS The qingyangshengenin content in the roots was significantly up-regulated, with the relative qingyangshengenin content in the roots being approximately 73.10 times higher than that in the leaves and 19.05 times higher than that in the stems. Transcriptomic analysis revealed that 269 DEGs annotated C₂₁ steroidal biosynthetic pathways. By analyzing the DEGs annotated between the comparison groups, 18 key enzymes were screened out in the C₂₁ steroidal synthesis pathway, which were encoded by 87 genes, among which AACT and other enzymes were the key enzymes in the upstream stage of the biosynthesis pathway. Quantitative real-time PCR was performed to verify the expression trend of eight DEGs, which was consistent with the corresponding transcriptome data. CONCLUSION The biosynthetic pathway of C₂₁ steroidal compounds is systematically analyzed in this study, and several key enzymes and coding genes are screened, which enrich the omics data of C. otophyllum. These findings lay a foundation for further study on the biosynthesis mechanism of C₂₁ steroid compounds of C. otophyllum.

OBJECTIVE To examine the herbal evidence of “Agaru” Tibetan medicines,conduct biopharcological studies and proveide a basis for the corrent origin and quality control. METHODS Through literature research, combined with field visits and expert consultations, variety classification, origins, characteristics and effects, compound preparation,and quality standards of “Agaru” type Tibetan medicines were compiled and summarized. The morphological and microscopic characteristics of “Aerma” medicinal materials were observed. Physicochemical identification was carried out based on thin-layer chromatography and determination of volatile oil components and content using GC-MS technology. RESULTS The name “Agaru” has evolved from Sanskrit loanwords and is a typical Tibetan medicine characterized by “multiple varieties and multiple origins”. Tibetan medicine often classifies it based on color into three categories: white (transliterated as “A Er Jia” or “A Jia Ga Bu”), black (transliterated as “A Er Na” or “A Ga Na Bao”), and red (transliterated as “A Er Ma” or “A Ga Ku Ao”). There is a significant difference in the varieties and origins of “Agaru” type Tibetan medicines used by Tibetan doctors in different regions, involving about 15 species (including varieties and forms) of plants from 5 families and 7 genera. Among them, “Alma” is widely used clinically, and its mainstream base is the heartwood of Camphora glandulifera (Wall.) Nees. Biopharmacological research was carried out on “Aerma” [C. glandulifera (Wall.) Nees], a clinically used variety of the “Agaru” group of Tibetan medicines, providing basic data for the establishment of its quality standards. CONCLUSION This study completes the botanical verification of the Tibetan medicine “Agaru” and the pharmacognostical research on “Aerma” [C. glandulifera (Wall.) Nees]. This will provide an important basis for the scientific evaluation of quality and in-depth development of “Agaru”.

OBJECTIVE To prepare methotrexate (MTX)-nicotinamide (NIC) coamorphous (MTX-NIC CA), evaluate its pharmacokinetic behavior in rats, and explore its solubilization mechanism. METHODS The coamorphous complex was prepared by melting method and characterized by powder X-ray diffraction and other techniques; the physical stability and thermodynamic stability of MTX-NIC CA were examined. The solubilization mechanism was studied by solubility profile method. The concentration of MTX in rat plasma was determined by high-performance liquid chromatography and the pharmacokinetic curve was drawn. RESULTS The physical stability of MTX-NIC CA was good; the reaction of MTX with NIC was a spontaneous enthalpy-driven reaction, and the mechanism of NIC for enhancing the solubility of MTX was that a soluble A_L-type complex with a ligand-to-metal ratio of 1∶1 was formed in solution. Compared with the raw drug MTX, the t_max of MTX-NIC CA was prolonged, and the c_max, AUC_0-t, AUC_0-∞ and F were significantly increased (P<0.05). CONCLUSION The MTX-NIC CA is successfully prepared by melting method with good stability, and the formation of co-amorphous complex improves the bioavailability of poorly soluble drugs.

OBJECTIVE To evaluate the safety characteristics of Rehmannia glutinosa leaf total glycosides capsules in juvenile Wistar rats, and provide reference for the use of Rehmannia glutinosa leaf total glycosides capsules in child and adolescent patients. METHODS Twenty-seven-day-old Wistar rats were randomly divided into control and low-, medium-, and high-dosage groups. Each dosage group was given the Rehmannia glutinosa leaf total glycosides by gavage repeatedly for 8 weeks, followed by a 4 week recovery. The clinical symptoms, body weight, food consumption, hematological and serum biochemical indexes, central nervous system function, learning and memory ability, skeletal development, reproductive function, organ weight and histopathological changes of the rats was observed. RESULTS Rehmannia glutinosa leaf total glycosides capsules did not show significant effects on the clinical symptoms, body weight, food consumption, hematological and serum biochemical indexes, main organ weights, central nervous system function, bones, sexual cycle, sperm counts, vitality and form of the rats. No histopathological changes were observed associated with Rehmannia glutinosa leaf total glycosides capsules. CONCLUSION The no-observed-adverse-effect-level (NOAEL) for Rehmannia glutinosa leaf total glycosides capsules in juvenile rats is determined to be 750 mg·kg^-1 in the 8 week feeding study. The data provides reference for the use of Rehmannia glutinosa leaf total glycosides capsules in child and adolescent patients.

OBJECTIVE To develop appropriate formulation and process design for hot melt extrusion (HME) of poorly soluble drug posaconazole with aid of rheology and discriminatory dissolution. METHODS The viscoelastic properties of polymer matrices were assessed for oscillation shear strain on a rotation disc rheometer within temperature of 14-180 ℃ and angular frequency of 100-0.1 rads·s^-1, respectively. A paddle method and an open flow cell method were developed alternatively to screen key critical quality attributes. RESULTS The selected polymer carrier showed storage modulus (G') >loss modulus (G″) with loss factor Tan(delta) <1 within the assessed temperature range, for better HME processability. Oscillation-frequency assessments further demonstrated that G'and G″ were more shear stable with angular strain at 140 ℃ compared with the increasing modulus trends at 150 and 160 ℃. Based on quality by design, discriminatory dissolution helped in defining if need to add excipient hydroxypropylcellulose in the process, as well as in designing HME granule size for formulation drug T. DSC, XRPD, Raman and optical microscopy characterization showed that the morphology of API changed from multicrystalline state to amorphous molecule dispersion after extrusion. CONCLUSION The drug release in vitro and in vivo of formulation drug posaconazole T is in bioequivalence with that of reference listing drug.

OBJECTIVE To prepare berberine hydrochloride microemulsion gel patch to improve the in vitro transdermal penetration and relative bioavailability in vivo. METHODS Firstly, through the screening of microemulsion dressing materials, and using the central point design response surface optimization method to optimize the microemulsion prescription, the best microemulsion prescription was obtained. The morphology and particle size of microemulsion were investigated by laser particle size analyzer and transmission electron microscope. Secondly, the matrix was selected for the gel paste, and then the optimal design of the gel paste was obtained by using the star design response surface optimization method. Finally, the microemulsion was added to the gel paste to prepare the berberine hydrochloride microemulsion gel paste, and the prescription process was verified. Through the in vitro transdermal test of mice, by comparing the permeation promoting effect of different concentrations of zzone, draw the cumulative permeation amount time curve of azone with different concentrations, and select the best amount of Azone. Finally, pharmacokinetic studies in rats and pharmacodynamic studies in mice were conducted. RESULTS Berberine hydrochloride microemulsion gel paste was successfully prepared. The best formulation of microemulsion was oil phase-emulsifier-co-emulsifier=0.11∶0.6∶0.3. The morphology and particle size of microemulsion were investigated by laser particle size analyzer and transmission electron microscope. The results showed that most microemulsion morphology was round, regular spherical, no aggregation, and the particle size was appropriate. The best prescription of gel paste NP700∶glycerol-dihydroxyaluminum aminoacetate=2.3∶17.5∶0.1 aluminum glycinate. The results of in vitro skin penetration test showed that azone with a mass fraction of 3% had the best penetration promoting effect. Pharmacokinetics showed that berberine hydrochloride microemulsion gel patch could prolong the action time in vivo. Preliminary pharmacodynamics shows that the drug can effectively improve the skin lesions, obviously inhibit the increase of inflammatory factors and improve the pathological tissue. CONCLUSION Microemulsion combined with gel patches can be used to prepare microemulsion gel patch with high drug loading and good therapeutic effect. The preparation of berberine hydrochloride microemulsion gel patch can effectively improve the bioavailability of berberine hydrochloride through percutaneous absorption.

OBJECTIVE To establish a population pharmacokinetic (PK) model for rosuvastatin and investigate the effects of demographic data, clinical characteristics, and genetic polymorphisms (ABCG2, SLCO1B1, SLCO1B3, SLCO10A1, ABCB1, CYP2C9) on its PK parameters in Chinese population. METHODS A total of 944 steady-state concentration data were collected from 944 patients with hyperlipidemia. UPLC-MS/MS was used to determine the plasma concentration of rosuvastatin. DNA was extracted and measured for concentration, and the Sequenom MassArray technology platform was utilized for genetic typing. Eventually a population PK model was established with non-linear mixed-effects modeling software (NONMEM). RESULTS The population estimates for the apparent clearance, apparent volume of distribution and absorption rate constant were 253 L·h^-1, 1 810 L and 0.318 h^-1, respectively. The datasets were best described by a one-compartment model with first-order elimination. The steady-state concentration of rosuvastatin increased as estimated glomerular filtration rate(eGFR) decreased and the variant allele for rs2199936 increased, under the same dosage regimen. Carrying one or two variant alleles for the ABCG2 rs2199936 showed a decrease of 32.6% and 53.2% in apparent clearance relative to the value in individuals without the variant allele. CONCLUSION Both ABCG2 rs2199936 and the eGFR were found to be significant covariates for apparent clearance. The results suggest that Chinese individuals with renal impairment and one or two variant alleles for the rs21999336 polymorphism (ABCG2) who are undergoing coronary angiography (CAG) should avoid high doses of rosuvastatin.

OBJECTIVE To explore the characteristics of the inhibitory effect of low concentrations of deguelin on the proliferation of activated T cells. METHODS Human peripheral blood mononuclear cells were isolated by Ficoll-Hypaque density gradient centrifugation, human T cells were purified using immunomagnetic beads, and T cells were activated with Anti-CD3/CD28 antibodies. Flow cytometry was used to detect T cell survival rate, proliferation index, apoptosis progression, CD25 expression level, and cell division ratio; ELISA was used to detect cytokines IL-2, IL-4, IL-6, IL-17, and IFN-γ secretion levels. RESULTS Low concentrations of deguelin inhibit T cell proliferation activated by anti-human CD3/CD28 antibodies, with an IC₅₀ of (73±12) nmol·L^-1, and a concentration of 400 nmol·L^-1 has no cytotoxicity. Low concentrations of deguelin do not affect the expression of CD25 and secretion of IL-2 in activated T cells but increases the proportion of G₀/G₁ phase cells. Low concentrations of deguelin promote the secretion of anti-inflammatory cytokine IL-4 and inhibit the production of pro-inflammatory cytokines IL-6, IL-17, and IFN-γ in T cells. CONCLUSION Low concentrations of deguelin significantly inhibit T cell proliferation in the G₀/G₁ phase and effectively suppress the secretion of pro-inflammatory cytokines, suggesting its potential role in the treatment of autoimmune diseases.

OBJECTIVE To study the changes of main components of Angelica sinensis(A.sinensis) after stir-frying high performance liquid chromatography(HPLC) fingerprint combined with chemometrics. METHODS The HPLC fingerprints of A. sinensis before and after stir-frying were established. The fingerprints were analyzed and evaluated by similarity evaluation,principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA).The variable importance in the projection(VIP)value> 1 was used as the standard to screen the different components before and after stir-frying of A. sinensis,and paired t-test was performed on the different components. RESULTS There were 20 common peaks in the fingerprints of raw A. sinensis(RAS)and stir-fried A. sinensis(SFAS). After comparison with mixed reference substances, eight chromatographic peaks were identified as tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, coniferyl ferulate, ligustilide, and butyliden phthalide.PCA(SFAS)showed that there were specific regions in the spatial distribution of principal components in the samples of RAS and SFAS. OPLS-DA screened out three differential components of ligustilide,senkyunolide I and coniferyl ferulate with VIP value> 1 as the standard.The peak areas of ligustilide,senkyunolide I and coniferyl ferulate were significantly reduced by paired t test (P<0.001). Chemometric analysis could effectively distinguish RAS and SFAS. CONCLUSION The chemical composition of RAS changes to a certain extent after stir-frying with soil.The established HPLC fingerprint had high similarity and could not significantly distinguish between RAS and SFAS. When combining HPLC fingerprint with stoichiometric analysis PCA,OPLS-DA and differential component distribution t test, RAS and SFAS can be obviously distinguished,which provides a reference for studying the processing principle of SFAS as well as data support for subsequent studies on pharmacological effects before and after processing.

OBJECTIVE To establish a quality control method of human chain-activated immune cell preparations. METHODS The ability to secrete cytokines and in vitro cytotoxicity were detected by co-culture and flow cytometry analysis. Purity was determined by flow cytometry. The concentration of live cells and cell viability were determined by AO/PI dual fluorescence cell counter. The residual cytokines were determined by enzyme-linked immunosorbent assay. Mycoplasma nucleic acid was detected by probe real-time PCR.Other detection items were carried out according to the provisions of the 3rd part of Chinese Pharmacopoeia 2020. RESULTS The detection of human chain-activated immune cell preparations was carried out using the established method,and the results of biological activity research, physical and chemical characteristics research, and cytokine residual detection were normally distributed. The detection results were valid,and the coefficient of variation(CV) was less than 20%. All other indicators met the requirements of the Chinese Pharmacopoeia 2020. CONCLUSION A preliminary quality control method of human chain-activated immune cell preparations are established, which has the characteristics of ensuring the quality control lability, safety, and effectiveness of the cell preparations. It can be used for the quality control of the human chain-activated immune preparations.

OBJECTIVE To investigate the clinical efficacy of aspirin combined with Huoxue Quyu decoction in the treatment of lower extremity arteriosclerosis obliterans (ASO). METHODS A total of 136 patients with lower extremity ASO admitted to our hospital were selected as the research objects, aged 49-87 years, with an average age of (65.00±6.54) years. According to treatment methods, 68 cases of conventional aspirin treatment were included in the control group, and 68 cases of Huoxue Quyu prescription combined with aspirin were included in the study group. The platelet function, hemodynamic index, blood lipid index, arteriosclerosis index, health status and clinical effect of the two groups were compared before and after treatment. Generalized estimation equation (GEE) model was used to analyze the factors affecting the clinical efficacy of ASO in the lower extremities, and GEE model was used to analyze the interaction effects of ankle-brachial index(ABI) at different time points, different groups and different stages. RESULTS After 6 weeks of treatment, the platelet function, hemodynamic indexes, lipid indexes and arteriosclerosis indexes of the two groups were significantly different compared with those before treatment (P<0.05). PAR, D-D, FIB, HWBV, LWBV, ESR, TC, TG and LDL-C levels in the study group were significantly lower than those in the control group (P<0.05), while BT, HDL-C, ABI, TBI, lameness distance, health status score and clinical efficacy were significantly higher than those in the control group (P<0.05). GEE model analysis showed that treatment method, treatment time and Fontaine stage significantly affected the therapeutic effect of ASO in lower limbs (P<0.05). In the interaction effect analysis, after 2 and 4 weeks of treatment, the ABI values of ASO patients in the study group and the control group were significantly different (P<0.05). After 6 weeks of treatment, different treatment methods had statistically significant effects on ABI values of ASO patients in stage Ⅱ, Ⅲ and Ⅳ (P<0.05). CONCLUSION Aspirin combined with Huoxue Quyu decoction is better than aspirin alone in the treatment of lower extremity ASO in the aspect of improvement of platelet function, hemodynamics, blood lipids and arteriosclerosis. After 6 weeks of treatment, the total effective rate and health status SF-36 score of the study group with combined medication are significantly higher than those of the control group. Treatment method, treatment time and Fontaine stage significantly affect the therapeutic effect of lower extremity ASO.

OBJECTIVE To discuss the problems needing attention in the pharmaceutical research of oral soluble film generic drugs, and to provide reference for the research and development of oral soluble film generic drugs. METHODS By searching the relevant foreign review reports and literature, this paper comparatively analyzed the prescription technology of ondansetron oral soluble film listed in different regions, and combined with the relevant guiding principles, discussed the general considerations in the development of oral soluble film. RESULTS The main research contents of ondansetron oral soluble film were determined around the concerns of prescription technology, quality control and stability. CONCLUSION In the development of oral soluble film generic drugs, clinical drug demand should be taken into consideration,combined with the characteristics of raw and auxiliary materials, prescription composition and production process,reasonable inspection items should be rationally established,and conducting in-depth pharmaceutical research to improve the quality of oral soluble film generic drugs.