Objective To observe the clinical effect and safety of recombinant human interferon-α2b(rhIFN-α2b)combined with 5-aminolaevulinic acid photodynamic therapy(ALA-PDT)in the treatment of patients with cervical intraepithelial lesions and human papilloma virus(HPV)infection.Methods The clinical data of patients with cervical intraepithelial lesions and HPV infection were analyzed retrospectively.The patients were divided into control group and treatment group according to cohort method.The control group was treated with rhIFN-α2b gel at posterior fornix,qd.On this basis,the treatment group was treated with ALA-PDT,namely applying 5％ALA temperature-sensitive gel prepared by 118 mg of ketone valerate hydrochloride powder for external use on vaginal surface,combined with photodynamic therapy,once a week.The clinical efficacy,time to recovery from clinical symptoms,recurrence rate,changes in cytokines[interleukin-6(IL-6),interleukin-8(IL-8)and interferon-γ(IFN-γ)]and immune function[T cell subsets CD3+,CD4+,CD8+and CD4+/CD8+],and safety were compared between the two groups.Results There were 98 cases in treatment group and 100 cases in control group.The total effective rates in treatment group and control group were 93.88％and 83.00％,with statistically significant difference(P＜0.05).The relief time of lower abdominal pain in treatment group and control group were(3.65±0.52)and(5.26±0.65)d;the time to recovery from abnormal vaginal discharge were(5.77±0.83)and(7.16±0.92)d;the time to recovery from irregular vaginal bleeding were(4.82±0.62)and(6.94±0.77)d;HPV clearance rates were 58.16％and 42.00％;IL-6 levels were(0.16±0.09)and(0.23±0.05)mg·L-1;IL-8 levels were(0.47±0.05)and(0.66±0.07)mg·L-1;IFN-γ levels were(10.07±0.98)and(7.24±0.65)ng·mL-1;CD3+were(71.06±8.29)％and(61.36±6.88)％;CD4+were(48.25±5.94)％and(42.25±5.13)％;CD8+were(20.37±2.42)％and(24.69±2.51)％;CD4+/CD8+were 2.11±0.27 and 1.36±0.16;the differences were all statistically significant(all P＜0.05).Recurrence rates in treatment group and control group were 5.10％and 14.00％,with statistically significant difference(P＜0.05).The total incidence of adverse drug reactions in treatment group and control group were 26.53％and 21.00％,without statistically significant difference(P＞0.05).Conclusion The combined treatment of rhIFN-α2b and ALA-PDT is effective for patients with cervical intraepithelial lesions and HPV infection.It has obvious advantages in improving immunity,relieving inflammatory response and reducing recurrence rate,with good safety.

Objective To investigate the effect of dapagliflozin-mediated miR-98-5p on high glucose-induced damage in human renal tubular epithelial cells.Methods Blood samples from patients with diabetic nephropathy(DN)and healthy individuals were collected.The expression of serum miR-98-5p was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR),and kidney injury-related indicators were measured using a biochemical immunoassay analyzer.HK-2 cells were cultured in vitro and randomly divided into control group(5 mmol·L-1 glucose),HG group(30 mmol·L-1 glucose),experimental-L group(30 mmol·L-1 glucose+20 μmol·L-1 dapagliflozin),experimental-H group(30 mmol·L-1 glucose+40 μmol·L-1 dapagliflozin),anti-miR-NC group(transfected with anti-miR-NC+30 mmol·L-1 glucose+40 μmol·L-1 dapagliflozin),and anti-miR-98-5p group(transfected with anti-miR-98-5p+30 mmol·L-1 glucose+40 μmol·L-1 dapagliflozin).Cell viability was evaluated using the cell counting kit 8(CCK-8)assay 24 hours after drug treatment;miR-98-5p expression in cells was detected by RT-qPCR;cell apoptosis rate was measured by flow cytometry,apoptosis-related protein expression was detected by Western blot;and inflammatory cytokine expression was measured by enzyme-linked immunosorbent assay.Results The expression levels of miR-98-5p in the serum of DN patients and healthy individuals were 1.00±0.25 and 0.39±0.05,respectively,showing a significant difference between the two groups(P＜0.05).The expression levels of miR-98-5p in the control group,HG group,experimental-H group,anti-miR-NC group,and anti-miR-98-5p group were 1.00±0.09,0.31±0.04,0.72±0.06,0.75±0.07 and 0.22±0.03;the cell survival rates were(100.00±3.36)％,(51.63±5.89)％,(79.46±9.90)％,(82.88±5.71)％and(59.69±7.43)％;apoptosis rates were(3.52±0.20)％,(35.80±3.67)％,(16.43±1.57)％,(15.71±1.42)％and(29.37±2.18)％;tumor necrosis factor-α(TNF-α)levels were(22.46±1.67),(68.37±6.05),(34.45±2.47),(35.11±2.84)and(60.46±3.56)pg·mL-1,respectively.The differences among these indicators were all statistically significant when comparing the HG group to the control group,the experimental-H group to the HG group,and the anti-miR-98-5p group to the anti-miR-NC group(all P＜0.05).Conclusion Dapagliflozin can alleviate high glucose-induced HK-2 cell damage by upregulating the expression of miR-98-5p,inhibiting inflammation,and reducing cell apoptosis.

Celastrol and wilforlide A are the main active triterpenoids of the traditional Chinese medicine Lei Gong Teng, which have anti-tumour, anti-inflammatory and immunosuppressive activities, and are the material basis for the clinical efficacy of Lei Gong Teng-related Chinese medicinal preparations. By analysing the biosynthetic pathway of active ingredients, optimizing genetic elements and utilizing "cell factory" to produce triterpenoids heterologously will be an effective way to obtain from Tripterygium wilfordii in the future in a "low-cost and high-efficient" manner. CYP712Ks are the first cytochrome P450s involved in the skeleton modification of friedelane-type and oleanane-type triterpenoids in T. wilfordii, and they can catalyse the generation of polpunonic acid from friedelin and 3-epi-katonic acid from β-amyrin by carboxylation at C-29 position. In this study, four multifunctional TwCYP712K1/2/3/5 were used to clarify the catalytic function and substrate selection preference using in vivo functional characterization in Saccharomyces cerevisiae. The spatial structure of the protein-substrate binding was clarified through homology modeling and molecular docking, and then the differential amino acids in the active pocket of the protein were mutated to clarify the crucial amino acids determining the catalytic function and the selection of substrate structure. A total of 63 mutant elements were constructed, and the amino acid sites affecting the carboxylation function of TwCYP712Ks were analyzed. In particular, the key amino acids affecting the substrate selectivity of TwCYP712K2 towards oleanane-type and friedelane-type triterpenoids were revealed and the TwCYP712K2^F127I and TwCYP712K2^A227T mutants would result in a reversal of the product ratio. In conclusion, four proteins of TwCYP712Ks were semi-rationally designed by homologous protein alignment and mutual mutation to elucidate multiple amino acid sites determining the catalytic function of the proteins, and a series of activity-enhancing or altering mutants were obtained, which provide abundant catalytic elements for the biosynthesis of active triterpenoids from T. wilfordii, and the mechanism of carboxylation in the C-29 position was initially elucidated.

The pathogenesis of depression is complex, and some existing monoamine antidepressants have problems such as drug resistance or off-target failure. Traditional Chinese medicine has the characteristics of "multi-component and multi-target", and has been used in the treatment of depression in clinical practice. Yueju pill is effective in the treatment of depression. Geniposide and ligustrazine, the active ingredients of Gardeniae fructus and Ligusticum sinense 'Chuanxiong', play a key role in the treatment of depression. In this study, based on the neuroprotective activity of genipin and the rapid antidepressant activity of tetramethylpyrazine, a series of novel genipin derivatives were designed and synthesized through pharmacophore assembly principle, and their neuroprotective activity and antidepressant effect were investigated. The results showed that the novel genipin derivatives had well neuroprotective activity on the glutamate-induced HT-22 cell model, with compounds W-1 and W-3 showing better protective activity. In behavioral despair depression (BDD) model mice, compound W-3 was found to have better antidepressant activity than W-1 in tail suspension test and forced swimming test. Further study on the behavior of chronic unpredictable mild stress (CUMS) model mice showed that W-3 could significantly improve the depression-like behavior of model mice. All animal experiments were approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine (approval number: AHUCM-mouse-2022027). The effects of the preferred compound W-3 on protein kinase A (PKA), cAMP response element binding protein (CREB), brain-derived neurotrophic factor (BDNF), 5-hydroxytryptamine 1A (5-HT_1A) receptor, N-methyl-D-aspartate ionic glutamate receptor 2A (GluN2A) and N-methyl-D-aspartate ionic glutamate receptor 2B (GluN2B) were analyzed by Western blot. W-3 treatment significantly up-regulated the protein expression of PKA, CREB, BDNF and 5-HT_1A, and down-regulated the protein expression of GluN2A and GluN2B. The results of qRT-PCR were consistent with those of Western blot. According to the above results, compound W-3 has a potential antidepressant effect, and its mechanism may be related to the activation of PKA-CREB-BDNF signaling pathway by regulating the expression of GluN2A, GluN2B and 5-HT_1A receptor proteins.

This experiment aims to study the taste-masking effects of different kinds of corrigent used individually and in combination on ibuprofen oral solution, in order to optimize the taste-masking formulation. Firstly, a wide range of corrigent and the mass fractions were extensively screened using electronic tongue technology. Subsequently, a combination of sensory evaluation, analytic hierarchy process (AHP)-fuzzy mathematics evaluation, and Box-Behnken experimental design were employed to comprehensively assess the taste-masking effects of different combinations of corrigent on ibuprofen oral solution, optimize the taste-masking formulation, and validate the results. The study received ethical approval from the Review Committee of the Beijing University of Chinese Medicine (ethical code: 2024BZYLL0102). The results showed that corrigent fractions and types were screened separately through single-factor experiments. Subsequently, a Box-Behnken response surface design combined with AHP and fuzzy mathematics evaluation was used to fit a functional model: Z = 688.310 11 - 3 023.722 22X₁ - 11.477 00X₂ + 62.721 67X₃ + 14.600 00X₁X₂ - 179.666 67X₁X₃ - 3.152 00X₂X₃ + 4 031.111 11X₁² + 0.525 28X₂² + 9.772 00X₃². This model is stable and reliable, determining the optimal taste-masking formulation to be 3.9 g·L^-1 of stevioside, 100 g L^-1 of xylitol, and 30 g L^-1 of methyl-β-cyclodextrin. The comprehensive score of the verification test is 88.14, with a relative RSD of 0.39%, indicating the feasibility of this model. This study achieved the improvement of the taste of ibuprofen oral solution through a combination of objective and subjective methods. Three types of corrigent were identified for enhancing drug taste, leading to the selection of the optimal taste-masking formulation for ibuprofen oral solution. This significantly enhanced the taste of the original formulation, improved patient compliance, and offered a new approach to mitigating the undesirable taste of formulations.

Three 2,3-diketoquinoxaline alkaloids were isolated from Heterosmilax yunnanensis Gagnep. Their structures were determined through 1D and 2D NMR, HR-ESI-MS, UV, and IR as 1-[5′-(3″-hydroxy-3″-methyl) glutaryl] ribityl-2,3-diketo-1,2,3,4-tetrahydro-6,7-dimethylquinoxaline (1), 1-[2′-(3″-hydroxy-3″-methyl) glutaryl]ribityl-2,3-diketo-1,2,3,4-tetrahydro-6,7-dimethylquinoxaline (2), and 1-ribityl-2,3-diketo-1,2,3,4-tetrahydro-6,7-dimethylquinoxaline (3). Compounds 1 and 2 are novel compounds, and 3 was isolated from H. yunnanensis for the first time. The hepatoprotective activity of these three compounds was evaluated, with compound 3 showing promising hepatoprotective activity.

Objective:To explore the genetic etiology of a fetus with Coffin-Siris syndrome2 (CSS2).Methods:A fetus with abnormal ultrasound findings detected at Luoyang Maternal and Child Health Care Hospital in July 2023 was selected as the study subject. Clinical data were analyzed retrospectively. Whole exome sequencing was carried out on fetal tissue and parental peripheral blood samples, and candidate variant was verified by Sanger sequencing and pathogenicity analysis. This study was approved by Medical Ethics Committee of the Luoyang Maternal and Child Health Hospital (Ethics No. LYFY-YCCZ-2023011).Results:Color Doppler ultrasound at 16 + gestational weeks revealed bilateral ventriculomegaly and cerebellar hypoplasia in the fetus. Trio-WES found that the fetus has harbored a heterozygous c. 553C>T (p.Gln185Ter) variant of the ARID1A gene, which was verified by Sanger sequencing to have a de novo origin. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c. 553C>T (p.Gln185Ter) variant of the ARID1A gene was classified as pathogenic (PVS1+ PS2+ PM2_Supporting). Conclusion:The fetus was diagnosed with CSS type 2, and the heterozygous c. 553C>T (p.Gln185Ter) variant of the ARID1A gene probably underlay its brain malformations.

Objective To study the pharmacokinetics and bioequivalence of two formulations of rasagiline mesylate tablets in healthy subjects under fasting and fed conditions.Methods The two-period,two-sequence,crossover study design was adopted in the fasting study.Thirty-six subjects were enrolled and given either test preparation or reference preparation 1 mg respectively in two periods.After collecting plasma samples,the plasma concentration of rasagiline was determined by liquid chromatography-tandem mass spectrometry(LC-MS/MS)and the bioequivalence was evaluated using the average bioequivalence(ABE)method.The four-period,two-sequence,fully replicate crossover study design was adopted in the fed study.Forty-eight subjects were enrolled and given the test preparation or the reference preparation at a dose of 1 mg twice respectively in four periods.According to the degree of intra-individual variation of Cmax,AUC0-t and AUC0-∞,the equivalence was evaluated using the reference-scaled average bioequivalence and ABE method,respectively.Results In the fasting study,the pharmacokinetic parameters of rasagiline of the test and reference preparation were as follow:Cmax were(9.70±3.14)and(9.62±3.85)ng·mL-1,AUC0-t were(6.03±1.47)and(6.02±1.95)ng·h·mL-1,AUC0-∞ were(6.13±1.51)and(6.12±1.97)ng·h·mL-1.The 90％confidence interval(CI)of the geometric mean ratio(GMR)were 94.11％-118.06％,99.22％-107.74％and 99.16％-107.44％for Cmax,AUC0-t and AUC0-∞,respectively,which were within the acceptance criteria of 80.00％-125.00％.In the fed study,the pharmacokinetic parameters of rasagiline of the test and reference preparation were as follow:Cmax were(3.00±1.92)and(3.52±1.77)ng·mL-1,AUC0_t were(5.02±1.20)and(5.06±1.20)ng·h·mL-1,AUC0-∞ were(5.11±1.23)and(5.14±1.22)ng·h·mL-1.The 90％CI of GMR were 96.99％-101.19％and 97.17％-101.41％for AUC0-t and AUC0-∞,which were within the acceptance criteria of 80.00％-125.00％.The 95％upper confidence bound of Cmax for were less than"0",and the point estimate of GMR were within the acceptance criteria of 80.00％-125.00％.The incidence of adverse events in fasting and fed studies was 22.86％and 22.92％,respectively,and all adverse events were moderate to mild.Conclusion The two rasagiline mesylate tablets were bioequivalent,and both the formulations were well tolerated.

Objective:To explore the association between frailty index (FI) and the risk for cardiovascular disease (CVD) in adults in Inner Mongolia Autonomous Region, and provide new evidence for the prevention of CVD in adults in Inner Mongolia Autonomous Region.Methods:The FI was constructed by using the data from a prospective cohort with a sample size of 25 055 individuals in 6 years of follow-up, and the prevalence of frailty in adults in Inner Mongolia Autonomous Region was described by the FI, and Cox proportional hazard regression model was used to evaluate the association between the FI and the incidence of CVD in adults in Inner Mongolia Autonomous Region.Results:The FI of the study population was 0.24±0.09. The population in the pre-frail (FI: 0.21-0.27) and frail (FI≥0.28) phases had increased risk for CVD compared to non-frail (FI≤0.20) population [pre-frail: hazard ratio ( HR)=1.232, 95% CI: 1.127-1.347; frail phase: HR=1.418, 95% CI:1.299-1.548]. For every 0.10 increase in FI, the risk for cardiovascular disease increased by 20.3% ( HR=1.203,95% CI:1.156-1.252). Conclusions:In this study, we constructed a FI, which can suggest the risk for CVD. As the increase of frailty degree, the risk for CVD increases.

Objective To explore the mechanism of melatonin(Mel)alleviating cholestatic liver injury in a mouse cholestasis model induced by cholic acid(CA)feeding.Methods A total of 15 8-week-old male C57BL/6J mice were randomly divided into control group,1％CA group,and 1％CA+Mel group,with 5 animals in each group.The control group was fed with normal chow diet,and the other 2 groups were fed with a diet containing 1％CA for 14 d to construct a model of cholestasis,and intraperitoneal injection with 100 mg/kg Mel was given to the mice from the 1％CA+Mel group.Immunohistochemical assay of α-SMA was applied for the liver tissues in the 1％CA group and the 1％CA+Mel group.The mRNA expression levels of fibrosis-related indicators in mouse liver tissue were examined by RT-qPCR.Liquid chromatography tandem mass spectrometry(LC-MS/MS)and automated biochemical analyzer were used to detect the contents of bile acids in the liver tissues and the serum of mouse,respectively.Then real-time qPCR and Western blotting were applied to detect the expression of bile acid synthesis and liver detoxification enzymes related indicators at mRNA and protein levels,respectively,to further investigate the mechanism of bile acid metabolism.Results Compared with the 1％CA group,the mRNA levels of liver fibrosis indicators(such as Tgfβ1,Col Ⅰ a1,Col Ⅱa1 and α-SMA)were significantly reduced(P＜0.05),and activation of stellate cells was obviously weakened displayed by immunohistochemical staining in the 1％CA+Mel group.The 1％CA+Mel group had notably decreased contents of bile acids in the serum and liver tissues,especially taurocholic acid and reduced mRNA levels of cholesterol 7α-hydroxylase(Cyp7a1)and Cyp8b1,while enhanced mRNA levels of hepatic detoxification enzymes Cyp2b10 and udp-glucuronosyltransferase(Ugt1a1)as well as protein levels of Cyp2b10 and sulfotransferase family 2A member 1(Sult2a1/2)when compared with the 1％CA group(P＜0.05).Conclusion Mel exerts its therapeutic effect on cholestasis by decreasing bile acid synthesis and increasing hepatic detoxification enzymes.