To observe the clinical efficacy and safety of tislelizumab injection in the treatment of patients with advanced non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) mutations and resistance to targeted therapy.

Patients with advanced NSCLC and EGFR mutations were assigned to control group and treatment group. The control group received 500 mg·m^-2 pemetrexed disodium for injection by intravenous infusion on the first day + 7.5 mg·kg^-1 bevacizumab injection by intravenous infusion every 3 weeks + 80 mg·m^-2 nedaplatin by intravenous infusion on the first day. On the basis of control group, the treatment group received 200 mg tislelizumab injection by intravenous infusion every 3 weeks. Two groups were treated for 3 cycles with 3 weeks per cycle. The clinical efficacy, tumor markers, quality of life and safety were compared between two groups.

Treatment group was enrolled 51 cases; control group was enrolled 51 cases. After treatment, the overall effective rates of treatment and control groups were 54.90% (28 cases / 51 cases) and 21.57% (11 cases / 51 cases), with statistically significant difference (P<0.01). After treatment, the levels of carcinoembryonic antigen (CEA) in treatment and control groups were (6.67±1.24) and (11.58±1.36) ng·mL^-1; the levels of cytokeratin-19 fragments were (7.64±1.47) and (12.25±1.63) ng·mL^-1; the levels of neuron-specific enolase were (22.65±2.81) and (25.05±2.94) ng·mL^-1; the quality of life scores were (73.67±6.24) and (64.45±5.36) points; the median progression-free survival were 7.00 and 10.00 months; the median overall survival were 14.00 and 20.00 months, respectively. The differences of above indexes were statistically significant between two groups (all P<0.05). The adverse drug reactions of two groups were nausea and vomiting, liver function impairment and renal function impairment. The incidences of total adverse drug reactions in treatment and control groups were 27.45% (14 cases/51 cases) and 31.37% (16 cases/51 cases), without statistically significant difference (P>0.05).

Tislelizumab injection has a definitive clinical efficacy in the treatment of patients with advanced NSCLC harboring EGFR mutations and resistance to targeted therapy, which can reduce the tumor marker levels, improve the quality of life, with high safety.

To explore the effects of psoralen and clopidogrel on metabolism of ciprofol in human liver microsomes.

A high performance liquid chromatography-tandem mass spectrometry method for the detection of ciprofol concentrations in human liver microsomes was established, and the specificity, linearity and lower limit of quantitation, precision, recovery rate, matrix effect and stability of this method were investigated. A series of remaining concentrations of ciprofol after co-incubation of with human liver microsomes were detected, and the results were substituted into GraphPad Prism 8 software to calculate the kinetic parameters of the enzymatic reaction of ciprofol in human liver microsomes. Afterwards, psoralen and clopidogrel were co-incubated with ciprofol, respectively, and the median inhibitory concentrations (IC₅₀) were calculated to investigate the effects of the two cytochrome P450 2B6 (CYP2B6) inactivators on ciprofol metabolism.

Ciprofol was linear in the range of 1.00-100.00 μg·mL^-1. The standard curve equation was y=0.15x-0.11 (r=0.999 2); the lower limit of quantification was 1.00 μg·mL^-1. The relative standard deviation (RSD) values for intra-day and inter-day precision were less than 7%, with relative error not exceeding ±5%. The extraction recoveries and matrix effects were 92.69%-99.62% and 94.77%-101.84%, respectively. In addition, the samples exhibited satisfactory stabilities incubated in a water bath at 37 ℃ for 2 h, stored at room temperature for 8 h or processed and placed in the autosampler for 24 h. The Michaelis constant (K_m), maximum reaction rate (V_max) and intrinsic clearance (CL_int) of ciprofol were 37.81 μg·mL^-1, 41.54 ng·min^-1·mg protein^-1 and 0.91 mL·min^-1·mg^-1, respectively. Psoralen and clopidogrel inhibited the metabolism of ciprofol in human liver microsomes in a dose-dependent manner with IC₅₀ values of 30.25 and 2.09 μmol·L^-1, showing mild and moderate inhibition of the metabolism of ciprofol, respectively.

Psoralen and clopidogrel can reduce the metabolism of ciprofol in human liver microsomes.

To explore the effects of Wu Wei Han bamboo slips “Yufang” on kidney injury in spontaneously hypertensive rats and its mechanism of action.

To analyze the differential genes between normal kidney tissues and hypertensive kidney tissues in GSE37460, a hypertensive kidney disease dataset in GEO database, and to explore the mechanism of hypertensive kidney injury; Spontaneously hypertensive rats (SHR) were randomly divided into model group (equal volume of distilled water), control group (7.59 mg·kg^-1 valsartan) and experimental-L, -M, -H groups (7.15, 14.29, 28.58 g·kg^-1 “Yufang” granules), with 10 rats in each group; additionally, Wistar-Kyoto (WKY) rats were taken as a normal group (equal volume of distilled water). Six groups were gavaged once a day for 8 consecutive weeks. The levels of urinary microalbumin (mAlb), N-acetyl-β-D-glucosaminidase (NAG) and serum thromboxane (TXB2), 6-keto-prostaglandin F1 α (6-keto PGF1α) were detected by enzyme linked immunosorbent assay. Western blot was used to detect the expression levels of hypoxia inducible factor-1 α (HIF-1α), vascular endothelial growth factor (VEGF) and transmembrane receptor protein 1 (Notch1) in rat kidney.

The data set GSE37460 contained 155 differential genes, 94 of which were up-regulated and 61 were down regulated and pathway enrichment involves pathways such as HIF-1α/VEGF/Notch1. The urinary mAlb levels of normal, model, control and experimental -H groups were (60.83±3.81), (159.60±20.65), (70.45±3.97) and (79.72±9.01) μg·L^-1, respectively; the urinary NAG levels were (34.37±6.92), (91.30±9.05), (44.65±10.09) and (54.79±2.87) ng·L^-1, respectively; the contents of TXB2 in serum were (78.58±3.78), (107.80±5.16), (80.92±4.02) and (85.65±2.17) pg·mL^-1, respectively; the serum 6-keto PGF1α levels were (121.50±3.45), (85.05±2.92), (108.30±2.96), (101.90±3.08) pg·mL^-1, respectively; the relative expression levels of HIF-1 α protein in rat kidney were 0.56±0.03, 1.25±0.02, 0.77±0.01 and 0.83±0.02, respectively; the relative expression levels of VEGF protein were 0.24±0.02, 1.24±0.03, 0.81±0.04 and 0.95±0.02, respectively; the relative expression levels of Notch1 protein were 0.33±0.03, 1.02±0.01, 0.50±0.04 and 0.68±0.01, respectively; there were significant differences in the above indexes between the control and experimental -H groups and the model group (all P<0.05).

The Wuwei Han bamboo slips “Yufang” can improve hypertension-related kidney damage and reduce kidney fibrosis, and its mechanism may be related to the inhibition of the HIF-1α/VEGF/Notch1 signaling pathway.

To explore the mechanism of long non-coding RNA prostate cancer-associated transcript 19 (LncRNA PCAT19) knockdown on oxaliplatin (OXA) sensitivity of human gastric cancer cells by regulating microRNA (miR)-200c-3p mediated ataxia-telangiectasia mutated proteins (ATM) and Rad3-related kinase-related kinase (ATR) / checkpoint kinase1(CHK1) signaling pathway.

Human gastric adenocarcinoma cells (AGS) were randomly divided into OXA group (8 μg·L^-1 OXA), OXA+si-NC+inh-NC group (si-NC and inh-NC co-transfected with +8 μg·L^-1 OXA), OXA+si-PCAT19+ inh-NC group (si-PCAT19 and inh-NC co-transfected +8 μg·L^-1 OXA), OXA+si-PCAT19+ miR-200c-3p inh group (si-PCAT19 and miR-200c-3p inh co-transfected +8 μg·L^-1 OXA). Cell proliferation in each group was detected by 5-acetylidene-2′-deoxyuridine assay; cell apoptosis was detected by in situ terminal transferase labeling technique; cell migration was detected by cell scratch assay; the expression levels of phosphorylated histone H2A.X (γH2A.X) and radiation-sensitive protein 51 (RAD51) were detected by immunofluorescence.

In OXA group, OXA+si-NC+inh-NC group, OXA+si-PCAT19+inh-NC group, OXA+si-PCAT19+miR-200c-3p group, the cell proliferation rates were (58.83±6.52)%, (59.42±6.86)%, (45.59±6.09)% and (55.71±7.69)%, respectively; the apoptosis rates were (20.06±2.44)%, (20.32±2.66)%, (31.74±4.43)% and (24.60±3.29)%, respectively; the migration rates were (54.50±6.14)%, (56.11±6.72)%, (31.51±4.13)% and (48.59±7.26)%, respectively; the relative fluorescence intensities of γH2A.X were 1.00±0.08, 0.97±0.12, 4.84±0.82 and 2.11±0.41, respectively; the relative fluorescence intensities of RAD51 were 1.00±0.06, 1.08±0.15, 0.22±0.04 and 0.55±0.08, respectively. There were no significant differences between OXA+si-NC+inh-NC group and OXA group (all P>0.05). The above indexes of OXA+si-PCAT19+inh-NC group were compared with those of OXA+si-NC+inh-NC group, the above indexes in OXA+si-PCAT19+miR-200c-3p inh group were compared with those of OXA+si-PCAT19+inh-NC group, the differences were statistically significant (all P<0.05).

Knockdown of LncRNA PCAT19 can affect the proliferation, migration, apoptosis and deoxyribonucleic acid repair of gastric cancer cells by targeting and negatively regulating miR-200c-3p, thereby enhancing the sensitivity of gastric cancer cells to OXA, which may be related to the ATR/CHK1 signaling pathway.

To investigate the effect of curcumin on lung injury in rats with acute organophosphorus pesticide poisoning (AOPP) through the cytochrome C/apoptotic protease activating factor-1 (CytC/Apaf-1) pathway.

SD rats were divided into control group, model group, inhibitor group, experimental -L and experimental -H groups, with 10 rats in each group. Except for the control group, the rats in the other 4 groups were administrated with 20 mg·kg^-1 oxomethoate to establish the rat model of organophosphorus pesticide poisoning. After the modeling was successful, the rats in the experimental -L and experimental -H groups were given 100 and 200 mg·kg^-1 curcumin by intragastric administration, and the rats in the inhibitor group were given 67.8 mg·kg^-1 YZ-488 intramuscularily, and the model group and control group were given equal volume of normal saline in the same way by intragastric administration and intramuscularily. Five groups of rats were given the drug once a day for 28 days. Mitochondrial membrane potential detection kits were applied to detect mitochondrial membrane potential (MMP). Enzyme-linked immunosorbent assay method was applied to detect levels of serum interleukin-10 (IL-10); apoptosis in lung tissue was detected by TUNEL; Western blot was applied to detect the expression levels of CytC and Apaf-1 proteins in lung tissue.

The MMP of experimental -L and experimental -H groups, control group, model group, and inhibitor group were (0.64±0.05), (0.78±0.09), (0.93±0.06), (0.51±0.04) and (0.76±0.08) RFU·mg^-1, respectively; IL-10 levels were (67.58±6.82), (104.29±11.37), (148.62±13.54), (32.84±3.51) and (98.74±8.76) pg·mL^-1, respectively; the apoptosis rates were (29.37±3.28)%, (16.58±1.87)%, (5.72±0.64)%, (43.68±4.91)% and (17.32±2.14)%, respectively; the relative expression levels of CytC protein were 0.89±0.07, 0.63±0.05, 0.42±0.04, 1.16±0.12 and 0.65±0.06, respectively; the relative expression levels of Apaf-1 protein were 0.73±0.06, 0.48±0.04, 0.32±0.03, 0.97±0.08 and 0.49±0.05, respectively. The above indexes of the model group were statistically significant compared with those in the experimental -L and experimental -H groups, inhibitor group and control group (all P<0.05).

Curcumin may reduce inflammatory response and oxidative stress in rats by inhibiting the CytC/Apaf-1 pathway, alleviate lung tissue and mitochondrial structural damage, and thereby improve lung injury in rats with AOPP.

To investigate the effect of Jinfu’an decoction on the apoptosis of non-small cell lung cancer cells (A549) by regulating phosphatidylinositol 3-kinase/ protein kinase B/ mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway mediated autophagy.

A549 cells were randomly divided into blank group, control group, combined group and experimental -L, -M, -H groups. The blank group was treated with 10% blank serum. The control group was treated with 20 μmol·L^-1 PI3K specific activator (740Y-P). The experimental-L, -M, -H groups were treated with 5%, 10% and 20% Jinfu ’an decoction containing serum, respectively. The combined group was treated with 20% Jinfu ’an decoction serum +20 μmol·L^-1 740Y-P. Flow cytometry was used to detect the apoptosis rate. Western blot was used to detect the expression levels of microtubule-associated protein 1A/1B -light chain 3 (LC3), sequestosome-1 (p62), Akt and mTOR.

The apoptosis rates of experimental -H group, combined group, control group and blank group were (31.48±1.35)%, (14.91±1.26)%, (3.30±0.45)% and (7.73±0.50)%, respectively; the relative expression levels of LC3 Ⅱ/Ⅰ protein were 1.13±0.10, 0.77±0.06, 0.31±0.04 and 0.50±0.04, respectively; the relative expression levels of p62 protein were 0.32±0.03, 0.56±0.05, 1.07±0.06 and 0.83±0.06, respectively; the relative expression levels of p-Akt/Akt protein were 0.33±0.04, 0.80±0.07, 0.98±0.09 and 0.52±0.03, respectively; the relative expression levels of p-mTOR/mTOR protein were 0.53±0.04, 0.84±0.06, 1.14±0.08 and 1.01±0.06, respectively. The above indexes of the experimental -H group were statistically different from those of the combination group and the blank group (all P<0.05).

Jinfu’an decoction can promote the apoptosis of A549 cells, and its mechanism may be related to the inhibition of PI3K/Akt/mTOR signaling pathway and the activation of cell autophagy.

To investigate the pharmacokinetic characteristics, safety and tolerability of single oral administration of trifluridine/tipiracil tablet in Chinese solid tumors under fasting and fed states, to evaluate the bioequivalence of the two formulations.

A randomized, open-label, two-formulation, two-sequence, four-cycle, repeated crossover design was used, with 30 subjects enrolled in both the fasting and fed groups, receiving a single oral dose of 60 mg of the test or reference formulation in each cycle. The concentrations of trifluridine and tipiracil in plasma were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Phoenix WinNonlin 8.3 and SAS 9.4 software were used for pharmacokinetic parameter and bioequivalence analysis.

Fasting group: The pharmacokinetic parameters of trifluridine were as follows. For the test preparation, C_max was (6 711.36±1 641.92) ng·mL^-1; AUC_0-t was (14 216.19±4 420.98) h·ng·mL^-1; AUC_0-∞ was (14 432.88±4 613.02) h·ng·mL^-1. For the reference preparation, C_max was (6 332.20±1 606.03) ng·mL^-1; AUC_0-t was (14 393.26±4 070.13) h·ng·mL^-1; AUC_0-∞ was (14 636.58±4 285.74) h·ng·mL^-1. The pharmacokinetic parameters of tipiracil were as follows. For the test preparation, C_max was (140.73±44.39) ng·mL^-1; AUC_0-t was (491.99±149.66) h·ng·mL^-1; AUC_0-∞ was (499.49±150.62) h·ng·mL^-1. For the reference preparation, C_max was (142.70±42.64) ng·mL^-1; AUC_0-t was (505.43±135.58) h·ng·mL^-1; AUC_0-∞ was (513.42±137.70) h·ng·mL^-1. Fed group: The pharmacokinetic parameters of trifluridine were as follows. For the test preparation, C_max was (3 703.22±875.59) ng·mL^-1; AUC_0-t was (10 817.63±3 466.63) h·ng·mL^-1; AUC_0-∞ was (10 986.12±3 776.68) h·ng·mL^-1. For the reference preparation, C_max was (3 754.66±915.72) ng·mL^-1; AUC_0-t was (10 775.42±3 594.37) h·ng·mL^-1; AUC_0-∞ was (11 002.36±4 106.89) h·ng·mL^-1. The pharmacokinetic parameters of tipiracil were as follows. For the test preparation, C_max was (53.73±16.72) ng·mL^-1; AUC_0-t was (219.63±74.00) h·ng·mL^-1; AUC_0-∞ was (226.61±77.00) h·ng·mL^-1. For the reference preparation, C_max was (54.73±14.42) ng·mL^-1; AUC_0-t was (224.66±75.23) h·ng·mL^-1; AUC_0-∞ was (231.91±78.74) h·ng·mL^-1. The geometric mean ratios and their 90% confidence intervals for the main pharmacokinetic parameters (C_max, AUC_0-t, AUC_0-∞) of the test and reference formulations of trifluridine/tipiracil were all within the range of 80.00%-125.00%. Common adverse drug reactions in the fasting group includes sinus bradycardia and elevated alanine aminotransferase, while common adverse drug reactions in the fed group included decreased lymphocyte count and detection of urinary sediment. The incidence of adverse drug reactions in the fasting and fed conditions were 36.67% (11 cases / 30 cases) and 46.67% (14 cases / 30 cases), respectively. There were no serious adverse events occurred during the trials in the both group.

The test and reference formulations of trifluridine/tipiracil tablet are bioequivalent under both fasting and fed conditions, with good safety and tolerability.