To study the effect of etanercept on retinopathy in diabetic rats and explore its mechanism.

Diabetic rat model was established by streptozotocin. The rats were randomly divided into control group (normal feeding), model group (diabetic rat model was established), experimental group (after modeling, etanercept 2 mg·kg^-1 was injected subcutaneously into the abdomen twice a week for 12 weeks), and combined group [based on the experimental group, 100 nmol·mL^-1 microRNA (miR)-30a inhibitor was injected into the tail vein]. The pathological morphology of retina was observed by hematoxylin-eosin staining. Enzyme-linked immunosorbent assay was used to detect the levels of serum inflammatory factors. The positive expression of glial fibrillary acidic protein (GFAP) in retinal of rats was detected by immunofluorescence staining. The relative expression level of miR-30a in retina was detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction.

In the model group, pathological changes occurred in each layer of retina, while in the experimental group, the pathological changes were alleviated, retinal edema was alleviated, cell arrangement was more regular, and the structural boundaries of each layer were clear. In control group, model group and experimental group, the serum levels of interleukin-6 (IL-6) were (0.42±0.07), (1.21±0.16) and (0.69±0.11) μg·L^-1, respectively; the levels of IL-1β were (0.26±0.05), (1.82±0.29) and (0.74±0.16) μg·L^-1, respectively; the levels of tumour necrosis factor-α(TNF-α) were (0.49±0.08), (1.25±0.17) and (0.72±0.13) μg·L^-1, respectively; the relative expression levels of GFAP in retinal tissues were 1.00±0.17, 3.18±0.56 and 1.45±0.23, respectively; the relative expression levels of miR-30a in retinal tissues were 1.00±0.19, 0.61±0.10, and 0.89±0.15, respectively; there were statistical differences between the model group and the control group, and between the experimental group and the model group (all P<0.05).

Etanercept can reduce the level of inflammatory factors in diabetic rats, and has a protective effect on diabetic rats retinopathy, its mechanism may be related to the regulation of miR-30a to activate insulin receptor substrate 1/phosphatidylinositol 3 kinase/protein kinase B signaling pathway.

To investigate the role of the receptor-interacting protein kinase 3 (RIP3)-mediated necroptosis pathway in penile tissue damage in diabetes mellitus-induced erectile dysfunction (DMED) rats and the effects of GSK872 and Yimusake intervention in DMED rats.

Male SD rats with normal sexual function were selected, and diabetic rats were constructed by 2 consecutive days injections of streptozotocin (45 mg·kg^-1), DMED rats were screened and randomly divided into model group, experimental group, Yimusake group and combined group. The normal and model groups were injected intraperitoneally with an equal amount of 0.9% NaCl and gavaged with an equal amount of distilled water; the experimental group was injected intraperitoneally with 1 mg·kg^-1 GSK872 and gavaged with an equal amount of distilled water; the Yimusake group was injected intraperitoneally with an equal amount of 0.9% NaCl and gavaged with 250 mg·kg^-1 Yimusake; the combined group was injected intraperitoneally with 1 mg·kg^-1 GSK872 and gavaged with 250 mg·kg^-1 Yimusake. The expression of RIP3, mixed lineage kinase domain like protein (MLKL) and transient receptor potential melastatin 7 (TRPM7) were detected by immunohistochemistry and immunofluorescence, α-smooth muscle actin (α-SMA) and Collagen Ⅰ were detected by Western blotting in rat penile tissues.

The positive area ratios of RIP3 in the penile tissues of normal, model, experimental, Yimusake and combined groups were (7.71±1.92)%, (35.72±2.73)%, (20.20±2.51)%, (19.94±2.66)% and (9.31±1.98)%; MLKL positive area ratios were (7.39±1.73)%, (26.48±1.74)%, (15.66±1.87)%, (15.88±1.59)% and (8.45±1.65)%; TRPM7 positive area ratios were (6.43±2.10)%, (35.77±1.78)%, (23.76±1.79)%, (25.24±2.08)% and (9.39±1.72)%; α-SMA protein relative expression levels were 1.53±0.13, 0.39±0.08, 0.88±0.14, 0.86±0.12 and 1.52±0.12; Collagen Ⅰ protein relative expression levels were 0.29±0.10, 1.20±0.14, 0.73±0.07, 0.71±0.11 and 0.31±0.10. The differences between the above indices in the model group compared with the normal group, the above indices in the experimental group compared with the model group, the above indices in the Yimusake group compared with the model group and the above indices in the combined group compared with the model group were all statistically significant (all P<0.05).

RIP3/MLKL/TRPM7 necroptosis pathway may participate in regulating the progression of DMED rats by promoting cell death and fibrosis, while GSK872 and Yimusake may ameliorate penile injury in rats by inhibiting this pathway.

To investigate the mechanism of Jianpi Yishentongqiao granule (JPYSTQG) in the intervention of spleen-kidney Yang deficiency type allergic rhinitis (AR) rats based on metabolomics.

The AR rat model of spleen-kidney-yang deficiency type was constructed by ovalbumin intraperitoneal injection sensitization + hydrocortisone intraperitoneal injection + rhubarb irrigation combined with modeling, and was randomly divided into model group and experimental group, with 10 rats in each group. Another 10 normal rats were selected as blank group. The experimental group was given 472.5 mg·mL^-1 JPYSTQG solution, the blank group and model group were given equal volume 0.9% NaCl. The rats in the three groups were administrated by gavage three times a day for 14 days. Serum levels of interleukin-4 (IL-4), immunoglobulin E (IgE) and interferon-γ (IFN-γ) were detected by enzyme-linked immunosorbent assay. Differential metabolites were screened by metabolomics and enriched by Kyoto Encyclopedia of Genes and Genomes.

The serum IgE levels in experimental, model and blank groups were (21.23±0.40), (23.80±0.67) and (20.03±1.42) ng·mL^-1; IL-4 levels were (111.00±5.86), (119.79±7.07) and (99.24±1.03) pg·mL^-1; IFN-γ levels were (1 980.63±15.63), (1 602.50±50.00) and (2 136.88±315.63) mg·mL^-1, respectively. Compared with the model group, the differences of experimental and blank groups were statistically significant (all P<0.01). Thirty-seven potential biomarkers of JPYSTQG intervention in spleen-kidney-yang deficiency rats were screened, mainly through arginine biosynthesis and tumor-related pathways.

JPYSTQG can regulate the expression of cytosine and other substances through arginine metabolism pathway, reduce IL-4, IgE and other inflammatory indicators, and thus have therapeutic effect on the allergic rhinitis rats with spleen-kidney-yang deficiency.

To observe the clinical efficacy and safety of caffeine citrate injection combined with poractant alfa injection in the treatment of respiratory distress syndrome (RDS) in premature infants.

RDS premature infants were divided into control group and treatment group according to the cohort method. The control group was given 100-200 mg·kg^-1 of poractant alfa injection by endotracheal tube injection and added 100 mg·kg^-1 after 12 h, whereas the treatment group received intravenous injection of caffeine citrate injection on the basis of the control group (with the initial loading dose of 20 mg·kg^-1 for 30 min, followed by an additional maintenance dose of 5 mg·kg^-1 for 10 min every 24 h). Both groups were treated continuously for 7 days. The clinical efficacy, treatment-related indicators, blood gas indicators, serological indicators and complications were compared between groups, and the safety evaluation was performed.

There were 57 cases in treatment group and 51 cases in control group. After treatment, the total effective rates in treatment group and control group were 94.74% (54 cases/57 cases) and 78.43% (40 cases/51 cases), respectively (P<0.05). After treatment, the mechanical ventilation time in treatment group and control group was (101.29±22.24) and (113.57±27.55) h; oxygen therapy time was (199.21±43.28) and (238.72±44.01) h; the hospitalization time was (25.63±3.37) and (29.98±4.90) d; the partial pressure of arterial blood oxygen (PaO₂) values were (79.55±7.27) and (74.83±6.16) mmHg; the arterial partial pressure of carbon dioxide (PaCO₂) values were (40.65±3.94) and (45.72±4.46) mmHg; blood pH values were 7.40±0.05 and 7.34±0.05; adrenocorticotropic hormone (ACTH) levels were (30.71±6.92) and (36.80±7.04) pg·mL^-1; cortisol (Cor) levels were (272.90±34.51) and (306.47±30.11) ng·mL^-1; interleukin-6 (IL-6) levels were (35.10±8.49) and (42.55±9.72) pg·mL^-1, and compared with control group, the above indicators in treatment group were statistically different (all P<0.05). During treatment, the incidence rates of complications in treatment group and control group were 3.51% (2 cases/57 cases) and 5.88% (3 cases/51 cases), respectively (P>0.05). There were no adverse drug reactions such as hypotension, bradycardia and irritability in both groups during treatment.

Caffeine citrate injection combined with poractant alfa injection in the treatment of RDS in premature infants can improve the efficacy, shorten the mechanical ventilation time, oxygen therapy time and hospitalization time, improve the blood gas and serum oxidative stress and inflammatory indicators, and the safety is good.

To observe the clinical efficacy and safety of rituximab injection combined with valsartan capsules in the treatment of patients with chronic glomerulonephritis.

The patients with chronic glomerulonephritis were randomly divided into control group and treatment group. The control group was given valsartan capsules 80 mg each time, qd, orally; on the basis of control group, the treatment group was combined with rituximab 1 000 mg per time, intravenous infusion, once on the 1st and 15th day. Two groups were treated for 12 months. The clinical efficacy, inflammatory indexes, renal function, disease recurrence and safety were compared between two groups.

Seventy-eight cases were enrolled in the treatment group, 9 cases were dropped out, and finally 69 cases were included in the statistical analysis; 79 cases were enrolled in the control group, 8 cases were dropped out, and finally 71 cases were included in the statistical analysis. After treatment, the total effective rates of treatment and control groups were 88.41% (61 cases/69 cases) and 61.97% (44 cases/71 cases), with significant difference (P<0.05). After treatment, the levels of interleukin-1 in the treatment and control groups were (73.88±9.91) and (86.47±10.27) pmol·L^-1, the levels of tumor necrosis factor-α were (42.17±5.66) and (47.94±5.52) pg·mL^-1, the levels of interferon-γ were (13.29±3.86) and (17.72±4.08) pg·mL^-1, the levels of creatinine were (82.17±12.23) and (87.89±10.21) μmol·L^-1, the levels of urea nitrogen were (4.66±1.26) and (5.71±1.65) mmol·L^-1, the 24-hour urinary protein quantifications were (3.55±1.12) and (4.69±1.57) g, the recurrence rates were 1.64% and 18.18%, and the differences were statistically significant (all P<0.05). The adverse drug reactions of treatment group were infection, neutropenia, nausea and vomiting, and leukopenia, while those in the control group were angioedema, nausea and vomiting, and neutropenia. The total incidences of adverse drug reactions in the treatment and control groups were 7.25% and 4.23%, without significant difference (P>0.05).

The clinical efficacy of rituximab injection combined with valsartan capsules in the treatment of patients with chronic glomerulonephritis is better than valsartan capsules alone, without increasing the incidence of adverse drug reactions.

To explore the mechanism of remimazolam improving learning and memory ability and brain injury in postoperative cognitive impairment (POCD) mice by regulating NOD-like receptor heat protein domain associated protein 3 (NLRP3).

The C57BL/6J mice were divided into sham group (exposure of the tibia only without truncation), model group (tibia ampution method to construct POCD model), oe-NC group (modeling+15 mg·kg^-1 remimazolam treatment+tail vein injection of 60 nmol·L^-1oe-NC), oe-NLRP3 group (modeling+15 mg·kg^-1 remimazolam treatment+tail vein injection of 60 nmol·L^-1oe-NLRP3), experimental-L, -M, -H groups (after modeling, 5, 10, 15 mg·kg^-1 remimazolam was respectively given intraperitoneal injection), 12 rats per group. For the treated mice, the Morris water maze and step - down test were used to evaluate their learning and memory abilities. The expression levels of NLRP3, interleukin-1β (IL-1β), IL-10 and tumor necrosis factor-α (TNF-α) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The level of reactive oxygen species (ROS) was detected by 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) staining, and the level of glial fibrillary acidic protein (GFAP) was detected by immunofluorescence.

The platform crossing times of sham operation group, model group and experimental-L, -M, -H groups were (4.58±0.91), (0.73±0.15), (0.92±0.18), (1.58±0.31) and (2.75±0.55), respectively; the error times were (0.50±0.11), (2.08±0.41), (1.75±0.35), (1.58±0.31) and (1.25±0.25), respectively; the relative expression levels of NLRP3 mRNA were 1.00±0.15, 3.51±0.69, 2.94±0.59, 2.57±0.51 and 1.91±0.38, respectively; the relative expression levels of IL-1β mRNA in sham operation group, model group, experimental-H group, control group and oe-NLRP3 group were 1.00±0.17, 2.72±0.54, 1.87±0.37, 1.91±0.38 and 2.66±0.53, respectively; the relative expression levels of IL-10 mRNA were 1.00±0.16, 0.51±0.10, 0.78±0.15, 0.73±0.15 and 0.56±0.11, respectively; the relative expression levels of TNF-α mRNA were 1.00±0.21, 3.08±0.61, 1.77±0.35, 1.82±0.36 and 2.65±0.53, respectively; the relative fluorescence intensities of ROS were 1.00±0.16, 7.22±1.45, 3.05±0.61, 3.13±0.63 and 6.75±1.31, respectively; the relative fluorescence intensities of GFAP were 1.00±0.18, 6.71±1.13, 3.38±0.63, 3.50±0.72 and 5.94±1.19, respectively. The above indexes in the sham operation group were compared with the model group, the above indexes in the experimental-H group were compared with the model group, the above indexes in the oe-NLRP3 group were compared with the control group, and the differences were statistically significant (all P<0.05).

Remimazolam improves postoperative cognitive function in mice by a mechanism that may be related to the inhibition of neuroinflammatory and oxidative stress damage by down-regulating NLRP3 expression.

To establish a quantitative analysis method for serum vitamin D (VD) based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and evaluate its clinical application value in pediatric nutritional support.

Serum samples were processed with acetonitrile, using 25-hydroxyvitamin D₂-d₆ (25-OH-VD₂-d₆) and 25-hydroxyvitamin D₃-d₆ (25-OH-VD₃-d₆) as internal standards. The chromatographic column was Kinetex C18 (50.0 mm × 3.0 mm, 2.6 μm). The mobile phase consisted of 0.1% formic acid aqueous solution and 0.1% formic acid methanol solution, with gradient elution at a flow rate of 0.5 mL·min^-1 and an injection volume of 10 μL. The column temperature was set to 30 ℃. Electrospray ionization was used in positive ion mode, with multiple reaction monitoring. The LC-MS/MS method was evaluated for specificity, calibration curves, lower limits of quantification, precision, recovery rate, stability, and matrix effects. The method was applied to detect VD levels in the serum of infants from the external group (oral 10 μg·kg^-1 VD drops, qd) and the parenteral group [parenteral nutrition solution with injectable fat-soluble vitamin (Ⅰ) administered intravenously, it was equivalent to 10 μg·mL^-1 VD, qd].

Both 25-OH-VD₂ and 25-OH-VD₃ showed good linearity in the range of 5-100 ng·mL^-1, with the 25-OH-VD₂ standard curve of y=1.00x+2.34×10^-2 (R=0.999 8), and a lower limit of quantification (LLOQ) of 0.5 ng·mL^-1; 25-OH-VD₃ standard curve of y=1.00x+5.6×10^-3 (R=0.999 8), and LLOQ of 1 ng·mL^-1. The recovery rate of 25-OH-VD₂ quality control samples was 95.59%-105.38%, with intra- and inter-day precision ≤ 4.75% and ≤ 4.68%, respectively. The recovery rate of 25-OH-VD₃ was 98.38%-113.63%, with intra- and inter-batch precision ≤5.64% and ≤7.83%, respectively. Stability of 25-OH-VD₂ and 25-OH-VD₃ samples at different concentrations was good under various conditions, with matrix effects ranging from 85%-110% and 88%-112%, respectively. After enteral or parenteral VD supplementation, the serum levels of 25-OH-VD₂ and 25-OH-VD₃ in both groups of preterm infants significantly increased, with the parenteral group showing significantly higher concentrations of 25-OH-VD₂ and 25-OH-VD₃ compared to the enteral group (P<0.05). The daily weight gain rate of preterm infants in the parenteral group was also significantly higher than those in the enteral group (all P<0.05).

The quantitative analysis method of serum VD based on LC-MS/MS has strong specificity and high sensitivity, which is suitable for the monitoring of VD level in premature infants.

To explore the design and application of digital intelligence quality control technology in drug clinical trials, aiming to improve the quality and efficiency of these trials.

Based on the characteristics and requirements of quality control in drug clinical trials, a digital intelligence quality control model was designed and applied throughout the entire process of drug clinical trials. The system’s performance and security were evaluated.

The digital intelligence quality management system can significantly enhance the accuracy and completeness of clinical trial data, improve the efficiency of drug clinical trials, and strengthen real-time monitoring capabilities to ensure compliance and safety of the trials.

The digital intelligence quality management system can optimize resource allocation, shorten trial duration, and reduce overall costs. It provides new perspectives and methodologies for quality control in drug clinical trials, laying a reliable foundation for the digital transformation of the biopharmaceutical industry.

To explore the omalizumab via microRNA (miR)-26a-5p reduced the inflammasome, or the NOD-like receptor themal domain associated protein 3 (NLRP3) mediated pyroptosis on airway remodeling in asthmatic rats, based on the inhibition of the Toll-like receptor 4 (TLR4)/myeloid differentiation primary response protein 88 (MyD88)/nuclear factor kappa-B (NF-κB) signaling pathway.

Fifty SD rats were divided into control group, model group, omalizumab group, omalizumab+agomir NC group and omalizumab+miR-26a-5p agomir group, with 10 rats in each group. The control group was intraperitoneally injected with 0.9% NaCl; the model group established the asthma model; the omalizumab group was intraperitoneally injected with 100 mg·kg^-1·d^-1 omalizumab daily on the basis of model group; the omalizumab + agomir NC group received tail vein injection of agomir NC on the basis of the omalizumab group; the omalizumab+ miR-26a-5p agomir group received tail vein injection of miR-26a-5p agomir on the basis of omalizumab group. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissue; enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors; real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and fluorescence in situ hybridization (FISH) were used to detect the expression of miR-26a-5p; Western blotting (WB) was used to detect the pyroptosis and TLR4/MyD88/NF -κB pathway-related proteins expression levels.

The tube wall thickness in control group, model group and omalizumab group were (35.16±2.95), (52.94±4.63) and (44.07±3.78) μm; interleukin-13 (IL-13) levels were (18.23±3.94), (61.84±10.94) and (34.23±6.11) pg·mL^-1; interferon gamma (IFN-γ) levels were (68.43±11.59), (22.94±4.70) and (43.10±8.04) pg·mL^-1; miR-26a-5p mRNA were 1.00±0.15, 2.26±0.31 and 1.37±0.19. The relative expression levels of NLRP3 in control group, the model group, the omalizumab group, the omalizumab +agomir NC group and omalizumab+miR-26a-5p agomir group were 1.00±0.11, 3.21±0.47, 1.77±0.24, 1.69±0.27 and 2.53±0.38; the relative expression levels of TLR4 were 1.00±0.13, 2.54±0.39, 1.86±0.39, 1.90±0.30 and 2.24±0.34; the relative expression levels of MyD88 were 1.00±0.15, 1.97±0.26, 1.44±0.21, 1.41±0.18 and 1.69±0.23; the relative expression levels of NF-κB were 1.00±0.12, 2.88± 0.37, 1.83±0.29, 1.77±0.25 and 2.54±0.41, respectively. The differences of the above indexes were statistically significant when compared the model group with the control group, the omalizumab group with the model group, and the omalizumab+miR-26a-5p agomir group with the omalizumab+agomir NC group (P<0.05, P<0.01).

Omalizumab may reduce NLRP3 inflammasome-mediated pyroptosis by inhibiting the activation of TLR4/MyD88/NF-κB pathway, thus achieving omalizumab inhibited airway remodeling and improved lung function in bronchial asthmatic rats.

To devolope a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of furmonertinib in human plsama and apply this method to analyze the blood concentration of non-small cell lung cancer (NSCLC) patients after taking medicine.

Furmonertinib -d3 was employed as the internal standard. The analyte and internal standard were extracted from plasma by protein precipitation with acetonitrile and chromatographed on Agilent Poroshell 120 SB-C₁₈(2.1 mm×100.0 mm, 3.5 μm); mobile phase consisted of acetonitrile-10 mmoL^-1 ammonium acetate(both of containing 0.1% formic acid); the whole analytical time was 3 min. The quantitative analysis were ionized with an electrospray ionization (ESI) source and operated in positive ion, multiple reaction monitoring (MRM) mode, the specificity, linear range, precision and accuracy, extraction recovery rate, matrix effect, dilution validation, and stability were all investigated.

The standard curves were demonstrated to be liner in the range of 0.50-100.00 μg·L^-1 with y=3.95×10^-2x-4.16×10^-4 (r=0.995 4). The accuracy was 95.8%-107.3%. The coefficient of variation (CV) of inter-day (n=6) and intra-day (n=3) for four different concentration levels were less than 15%. The average recoveries and matrix effects were between 83.1%-88.2% and 94.6%-105.8%.

The established LC-MS/MS method for determining the concentration of vormetinib meets the requirements of biological sample analysis and can be used for the concentration determination of vormetinib in clinical practice.