To establish an ultra-high performance liquid chromatography-tandem mass spectrometry method for the determination of 14 sedative hypnotics and their metabolites in serum and urine.

Serum or urine samples were treated with acetonitrile to precipitate proteins. Separated on an Agilent EclipsePlus C18 RRHD column, the liquid was eluted with 0.1% formic acid in water and methanol, at a flow rate of 0.30 mL·min^-1 and column temperature of 35 ℃. Quantified by the internal standard method, and multiple reaction monitoring (MRM) was performed by electrospray ionization (ESI⁺). The method was evaluated for specificity, standard curves and limits of quantification, precision and recovery, matrix effects, and stability. This method was applied to test the proficiency testing samples from the 2023 National Medical Emergency Toxicant Detection Laboratory Comparison Program, including qualitative and quantitative test samples for emergency poisoning assessment of sedative-hypnotic drugs in serum.

The method demonstrated good specificity, with no cross-interference between the various sedative hypnotics and their metabolites. The linear range of 14 sedative hypnotics and metabolites were 1.00-45.00 μg·L^-1, with a correlation coefficient greater than 0.998 2, the detection limit in serum and urine was 0.30 μg·L^-1, and the quantification limit was 1.00 μg·L^-1. In serum, recovery rates at three spiked levels (10.00, 50.00 and 200.00 μg·L^-1) ranged from 85.60% to 105.20%, with relative standard deviations (RSD) between 0.73% and 4.45%. In urine, recovery rates ranged from 88.93% to 108.12%, with relative standard deviations between 0.50% and 6.92%. Fluphenazine hydrochloride and clozapine were identified by this method. The two determined quantitative concentrations of fluphenazine hydrochloride were 339.00 μg·L^-1 and 349.00 μg·L^-1 , with an average value of 344.00 μg·L^-1 and a deviation of 2.90%. The two determined quantitative concentrations of clozapine were 1 668.00 μg·L^-1 and 1 707.00 μg·L^-1 , with an average value of 1 688.00 μg·L^-1 and a deviation of 2.30%. The final feedback result was satisfactory.

This method is rapid and accurate, making it suitable for emergency poisoning detection of sedative hypnotics. Its application to the blind sample detection of emergency poisoning cases is satisfactory.

To evaluate the predictive value of different risk models for hepatocellular carcinoma (HCC) in patients with chronic hepatitis B (CHB)-related cirrhosis receiving antiviral therapy with entecavir (ETV), tenofovir disoproxil fumarate (TDF), and tenofovir alafenamide fumarate (TAF), and to clarify the effects of pharmacological differences among various nucleos(t)ide analogs (NAs) on HCC risk, so as to provide a reference for clinical diagnosis and treatment.

A total of 252 treatment-naive patients with CHB-related cirrhosis without a history of HCC who received NA antiviral therapy from May 2015 to May 2020 were retrospectively enrolled. They were divided into the ETV group (n=108), TDF group (n=96), and TAF group (n=48) according to the NA used. Cox proportional hazards model was used to construct an HCC risk prediction model. Patients were classified into low-risk group (0-3 points), intermediate-risk group (4-7 points), and high-risk group (8-13 points) based on the Asia-Pacific Association for the Study of the Liver (APASL) REALB score. All patients continued to receive corresponding NA therapy and were followed up until the study endpoint. The predictive performance of each risk score was compared using time-dependent area under the receiver operating characteristic curve (AUROC), and the differences in HCC incidence among different NA groups were analyzed. The main outcome measures included HCC incidence, international normalized ratio (INR), alpha-fetoprotein (AFP), diabetes mellitus, drinking history, predictive performance of the REALB score, and prognosis-related effects of different NAs.

A total of 252 patients were screened, and finally 108 patients were assigned to the ETV group, 96 to the TDF group, and 48 to the TAF group. The median follow-up duration was 56.96 months, and hepatocellular carcinoma (HCC) was diagnosed in 19.00% (48 cases/252 cases) of patients. The 1year incidence rates of HCC in the ETV, TDF, and TAF groups were 4.63% (5 cases/108 cases), 5.21% (5 cases/96 cases), and 4.17% (2 cases/48 cases), respectively; the 3 year rates were 12.96% (14 cases/108 cases), 13.54% (13 cases/96 cases), and 12.50% (6 cases/48 cases), respectively; and the 5 year rates were 17.59% (19 cases/108 cases), 18.75% (18 cases/96 cases), and 16.67% (8 cases/48 cases), respectively. No statistically significant differences were observed among the three groups (all P>0.05). Multivariate stepwise regression analysis revealed that INR (HR=2.77, 95% CI: 1.46-5.25, P<0.01), AFP (HR=1.00, 95% CI: 1.00-1.00, P<0.05), diabetes mellitus (HR=3.06, 95% CI: 1.54-6.08, P<0.01), and history of alcohol consumption (HR=2.25, 95% CI: 1.04-4.86, P<0.05) were independent risk factors for HCC development. The predictive performance of the REALB score at 3 years (AUROC=0.74) and 5 years (AUROC=0.70) remained higher than that of other models except the RWSHCC model, while its 1year AUROC was similar to those of other models. Statistically significant differences in the 1, 3, and 5 year HCC incidence rates were found between the intermediaterisk group and the highrisk group (all P<0.001).

Different NAs (ETV, TDF, TAF) have comparable efficacy in controlling HCC risk in patients with CHB-related cirrhosis, and differences in their pharmacological properties do not significantly affect HCC risk. The REALB score shows stable and excellent predictive ability for HCC in patients receiving NA antiviral therapy and can be used as a preferred tool for clinical prognosis evaluation.

To explore the effects of Wilms tumor 1-associating protein (WTAP) rs7766006 polymorphisms on chemotherapy toxicities and clinical prognosis in children with brain tumors.

Pediatric patients with brain tumors who received chemotherapy at our hospital were included as study subjects. Matrix-assisted laser desorption/ionization time of flight mass spectrometry was used for WTAP rs7766006 genotyping. Clinical data collected included chemotherapy toxicities and tumor progression. The associations of WTAP rs7766006 G>T polymorphisms with chemotherapy toxicities and progression-free survival (PFS) were analyzed. The expression of WTAP in brain tumors and its prognostic significance, and the potential mechanism of rs7766006 G>T polymorphisms in WTAP expression were explored based on bioinformatics methods.

Among the 107 children with brain tumors included, the rs7766006 GG homozygous, GT heterozygous, and TT homozygous genotypes accounted for 40.19% (43 cases/107 cases), 44.86% (48 cases/107 cases) and 14.95% (16 cases/107 cases), respectively. The frequencies of G and T alleles were 62.62% (134 cases/214 cases) and 37.38% (80 cases/214 cases) respectively. The incidence rates of mucositis in the GG, GT, and TT genotype groups were 53.49% (23cases/43 cases), 27.08% (13 cases/48 cases) and 43.75% (7 caes/16 cases), respectively. The incidence rates of coagulation disorders in three groups were 18.61% (8 cases/43cases), 2.08% (1 case/48 cases) and 6.25% (1 case/16 cases), respectively. The difference in the incidence rates of the two chemotherapy toxicities mentioned above between the GG and GT genotypes was statistically significant (all P<0.05). However, there were no significant differences in the incidence of other chemotherapy toxicities among the three groups (all P>0.05). The disease progression rates for the GG, GT, and TT genotype groups were 65.12% (28 cases/43 cases), 43.75% (21 cases/48 cases), and 62.50% (10 cases/16 cases), respectively. The risk of disease progression in children with the GG genotype was significantly higher than in those with the GT genotype (P<0.05). Bioinformatics analysis showed that the WTAP expression in brain tumors 6.00±0.66 was significantly higher than that in normal tissues 4.63±1.34 (P<0.001). The median overall survivals for the WTAP high-expression group and the low-expression group were 537 and 2 835 days, respectively (P<0.001). The rs7766006 polymorphism was located in the exonic splicing enhancer site and possibly regulated WTAP expression by affecting alternative splicing.

WTAP rs7766006 GG genotype might be a risk factor for oral mucositis, coagulation disorders, and progression in children with brain tumors.

To analyze the strengths and weaknesses of China’s R&D in obesity/overweight drug clinical trials by comparing domestic and international trial characteristics, and put forward suggestions based on national conditions.

Clinical trial data of drugs for overweight/obesity were retrieved from ClinicalTrials.gov (CT.gov) and the China Drug Clinical Trial Registration and Information Disclosure Platform (Registration Platform), and divided into a domestic trial group and an international trial group. A bibliometric method was used for comparative analysis from the dimensions of drug type, dosage form, trial design, number of trials, age, gender, and sample size.

Domestically, chemical drugs dominated (77.57%, 211 cases/271 cases), while international trail chemical drugs (51.30%, 256 cases/499 cases) and biological products (48.30%, 241 cases/499 cases) developed balancedly. Domestic trials focused on tablets (46.32%, 126 cases/271 cases) and injections (47.06%, 128 cases/271 cases), whereas international trials were dominated by injections (57.52%, 287 cases/499 cases) (P<0.001). Domestic randomization rate (92.25%, 250 cases/271 cases) was much higher than overseas (79.56%, 397 cases/499 cases), but blinding application was lower (44.65%, 121 cases/271 cases vs. 65.33%, 326 cases/499 cases). Few domestic trials included children (1.11%, 3 cases/271 cases), with only 3 female-specific studies.

Global obesity/overweight drug trials are mainly in China and the US, but China lags behind with unbalanced structure and insufficient population representativeness. China should improve R&D level through policy incentives and international cooperation based on clinical needs.

To investigation the effect of arbutin on hypoxia/reoxygenation (H/R)-induced injury of cardiomyocytes (H9C2) through the neurogenic locus notch homolog protein 1(Notch1)/hairy and enhancer of split 1 (Hes1) signaling pathway.

An H/R-induced H9C2 cell injury model was established. The optimal intervention concentration of arbutin was screened by pretreating cells with various concentrations (12.50-200.00 μmol·L^-1). H9C2 cells were divided into a normal group, a model group, experimental-L, M, H groups (25.00, 50.00, and 100.00 μmol·L^-1 arbutin, respectively), and an inhibitor group {100.00 μmol·L^-1 arbutin+10.00 μmol·L^-1 N-[N-(3, 5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT)}. Cells in the normal group were cultured under normal conditions, while H/R models were established in the remaining groups. All treatment groups received arbutin pretreatment at concentrations ranging from 12.50 to 200.00 μmol·L^-1 for 24 hours before modeling. Cell proliferation was detected by the CCK-8 assay, and apoptosis was assessed by TUNEL assay; mitochondrial membrane potential was measured by JC-1; reactive oxygen species (ROS), lactate dehydrogenase (LDH), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and superoxide dismutase (SOD) levels were detected using kits. Expression of Notch1, Hes1, Bcl-2, Bax, and caspase-3 was detected by Western blot.

Arbutin at concentrations of 25.00–200.00 μmol·L^-1 significantly increased the survival rate of H/R-induced H9C2 cells (P<0.05). Arbutin at 25.00, 50.00, and 100.00 μmol·L^-1 was selected as the low, medium, and high doses for subsequent experiments. The creatine kinase isoenzyme MB (CK-MB) levels in the normal group, model group, experimental-H group, and inhibitor group were (2.45±0.39), (8.72±0.82), (3.56±0.42), and (5.48±0.57) U·mL^-1, respectively; the cTnI levels were (22.58±2.23), (68.34±5.96), (30.12±3.26), and (49.62±5.15) ng·L^-1, respectively; the LDH levels were (53.42±5.44), (163.72±13.41), (65.45±6.57), and (98.33±9.85) U·L^-1, respectively; the relative ROS levels were 1.01±0.12, 3.34±0.35, 1.46±0.18, and 2.24±0.23, respectively; the SOD levels were (96.23±8.12), (24.31±2.24), (78.39±7.93), and (43.78±4.49) U·mg^-1, respectively; the GSH-Px levels were (74.82±7.55), (16.42±1.83), (61.32±6.29), and (35.72±3.38) U·mg^-1, respectively; the MDA levels were (15.45±2.03), (58.33±5.57), (21.56±2.24), and (42.47±4.13) μmol·L^-1, respectively; the apoptosis rates were (2.45±0.39)%, (29.86±3.25)%, (9.86±0.91)%, and (21.34±2.25)%, respectively; the red/green fluorescence intensity ratios were (5.78±0.65)%, (1.42±0.20)%, (4.89±0.54)%, and (2.85±0.33)%, respectively; the Notch1 protein expression levels were 1.27±0.12, 0.34±0.03, 1.05±0.08, and 0.54±0.05, respectively; the Hes1 protein expression levels were 0.95±0.10, 0.25±0.03, 0.81±0.08, and 0.57±0.05, respectively; the Bcl-2 protein expression levels were 1.01±0.11, 0.31±0.03, 0.89±0.08, and 0.61±0.06, respectively; the Bax protein expression levels were 0.45±0.04, 1.34±0.12, 0.61±0.06, and 0.93±0.09, respectively; and the caspase-3 protein expression levels were 0.23±0.03, 0.91±0.09, 0.37±0.04, and 0.64±0.06, respectively. Comparisons between the normal group and the model group, between the model group and the experimental-L, M, H groups, and between the experimental-H group and the inhibitor group all showed statistically significant differences for the above indicators (all P<0.05).

Arbutin alleviates H/R-induced H9C2 cell injury by activating the Notch1/Hes1 pathway and inhibiting oxidative stress.

To explore the effect and safety of rituximab injection in the treatment of patients with primary membranous nephropathy (PMN).

PMN patients were selected as the study subjects and divided into control group and treatment group according to envelope method. Patients in control group received valsartan capsules, 80 mg·d^-1, and the patients in treatment group was treated with 375 mg·m^-2 rituximab injection, (once a week, and adjusted to once per month after one month of therapy) on the basis of control group. Both groups received continuous treatment for six months. The clinical efficacy, renal function indexes (24-hour urine protein, creatinine, blood urea nitrogen), immune cells (Th1 cells, Th2 cells, Th17 cells), immune-related factors [immunoglobulin G subtype 4 (IgG4), interleukin-4 (IL-4)], vascular endothelial cell function [plasminogen activator inhibitor-1 (PAI-1), endothelin-1 (ET-1)], soluble suppression of tumorigenicity 2 (sST2) and Toll-like receptor 4 (TLR4) levels were compared and analyzed and the safety were evaluated.

The clinical effective rates of the treatment group and the control group were 86.49% (64 cases/74cases) and 63.51% (47 cases/74 cases), respectively, and the difference was statistically significant (P<0.01). After treatment, the 24 h urine protein in the control group and the experimental group were (2.03±0.39) and (1.49±0.32) g·24 h^-1, respectively; the serum creatinine were (98.11±12.29) and (76.69±10.12) μmoL·L^-1, respectively; blood urea nitrogen was (7.39±0.89) and (6.08±1.27) mmoL·L^-1, respectively; the proportion of Th2 cells were (0.92±0.11) % and (0.76±0.13) %, respectively; the levels of sST2 were (1.59±0.38) and (1.19±0.31) ng·mL^-1, respectively; the levels of TLR4 were (125.09±13.91) and (91.58±10.67) ng·mL^-1; the levels of ET-1 were (53.58±5.34) and (41.77±4.35) pg·mL^-1. The differences of the above indicators between two groups were all statistically significant (all P<0.01). The drug adverse effects of treatment group and the control group were 16.22% (12 cases/74 cases) and 13.51% (10 cases/74 cases) (P>0.05).

Rituximab injection is beneficial for primary membranous nephropathy to improve renal function and vascular endothelial cell function, reduce the release of sST2 and TLR4, reduce the expression of immune-related factors, improve clinical efficacy, and have a good safety profile.

To explore the effects of woginostatin (WOG) on the biological behaviors of gastric cancer cells through long non-coding RNA small nucleolar RNA host gene 16 (LncRNA SNHG16)/microRNA (miR)-302a-3p/E2F transcription factor 1 (E2F1).

In cell experiment, human gastric cancer cells (MGC-803) were divided into cell control group (normal culture), cell experimental-H group (100 μmol·L^-1 WOG treated cells), oe-NC group (transfected oe-NC), oe-LncSNHG16 group (transfected oe-LncSNHG16), mimic-NC group (transfected mimic-NC), miR-302a-3p mimic group (transfected miR-302a-3p mimic), WOG + oe-NC group (transfected oe-NC based on 100 μmol·L^-1 WOG treated cells), WOG + oe-LncSNHG16 group (transfected oe-LncSNHG16 based on 100 μmol·L^-1 WOG treated cells), WOG +oe-LncSNHG16+miR-302a-3p mimic group (transfection of oe-LncSNHG16 and miR-302a-3p mimic on the basis of 100 μmol·L^-1WOG treatment group). In animal experiment, the subcutaneous transplanted tumor model of gastric cancer was established in BALB/C male nude mice. The nude mice were randomly divided into animal experimental-L, -M and -H dose groups (15, 30 and 60 mg·kg^-1 WOG gavage), and the animal control group was gavaged with equal volume of physiological saline, with 10 mice in each group. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of LncRNA SNHG16, miR-302a-3p and E2F1. The cell viability was detected using the cell counting kit 8 (CCK-8). The cell proliferation, invasion and apoptosis were detected using 5-ethynyl-2′-deoxyuridine (Edu), Transwell and in situ terminal transferase labeling technique (TUNEL).

In cell experiment, the relative expression levels of LncRNASNHG16 in cell control group, cell experimental-H group, mimic-NC group, miR-302a-3p mimic group, oe-NC group and oe- LncSNHG16 group were 1.00±0.13, 0.47±0.05, 1.00±0.10, 0.81±0.12, 1.02±0.09 and 3.76±0.41, respectively; the relative expression levels of miR-302a-3p were 1.00±0.11, 3.60±0.39, 1.00±0.09, 3.74±0.32, 0.97±0.10 and 0.27±0.03, respectively; the relative expression levels of E2F1 mRNA were 1.00±0.14, 0.23±0.02, 1.00±0.10, 0.33±0.03, 0.95±0.08 and 2.18±0.25, respectively. Compared the cell experimental-H group with cell control group, compared miR-302a-3p mimic group with mimic-NC group and compared oe-LncSNHG16 group with oe-NC group, the above indicators were all statistically significantly different (P<0.01, P<0.001). The proliferation rates of cell control group, cell experimental-H group, WOG + oe-LncSNHG16 group and WOG + oe-LncSNHG16 + miR-302a-3p mimic group were (95.04±10.14)%, (36.36±4.75)%, (97.64±11.04)% and (31.87±3.56)%, respectively; the number of invasive cells were (147.36±15.01), (43.31±6.25), (153.45±15.22) and (37.35±5.04), respectively; the apoptosis rates were (7.83±0.83)%, (35.77±4.38)%, (8.44±0.88)% and (29.35±3.09)%, respectively. There were statistically significant differences in the above indexes between the cell experimental-H group and the cell control group, between the WOG + oe-LncSNHG16 group and the cell experimental-H group, and between the WOG + oe-LncSNHG16 group and the WOG + oe-LncSNHG16 + miR-302a-3p mimic group (all P<0.001). In animal experiment, the tumor volume of animal control group and animal experimental-L,-M,-H groups were (2 554.50±280.41), (2 194.10±234.22), (1 512.40±166.75) and (1 146.20±124.32) mm³, respectively; the tumor mass was (2.13±0.28), (1.84±0.23), (1.65±0.21) and (1.32±0.14) g, respectively. There were statistically significant differences in the above indicators between experimental-L, -M, -H groups and animal control group (P<0.05, P<0.01, P<0.001).

WOG can inhibit the proliferation and invasion of gastric cancer cells and promote their apoptosis. This may be related to the regulation of the LncRNA SNHG16/miR-302a-3p/E2F1 axis, which blocks the cell cycle.