Objective: This study employed next-generation sequencing (NGS) for large-scale genotyping of 42 blood group systems in blood donors to evaluate its applicability in multi-system blood group identification and rare blood type repository construction, while exploring the complementary value of third-generation sequencing (TGS) in haplotype phasing and variant capture in regions with insufficient sequencing depth for ABO ambiguous samples. Methods: A total of 562 regular blood donors (median donation frequency 11 times) from a blood center were enrolled. NGS was used for genotyping of 42 blood group systems, with a focused analysis on 12 systems essential for rare blood type repository construction: MNS, Lutheran, Kell, Duffy, Kidd, Diego, Yt, Colton, Gerbich, Ok, H, and I. TGS was employed to resolve haplotype phasing (i. e., determining whether different mutation sites are located on the same chromosome) in 12 ABO samples with discrepancies between forward and reverse typing. Serological and genotyping results were compared for MNS, Duffy, Kidd, Lewis, and ABO systems. Results: NGS-based genotyping revealed that Fy (a-b+) in the Duffy system accounted for 0.87% (4/458), and Di (a+b+) in the Diego system accounted for 10.62% (31/292). The Lutheran, Kell, Yt, Colton, H, Ok, I, and Gerbich systems exhibited near-uniform phenotype distributions. Among the 12 ABO ambiguous samples, NGS yielded multiple possible genotype combinations in 5 cases due to inability to phase alleles, whereas TGS uniquely resolved the genotypes through long-read sequencing. In one case, NGS detected only 2 mutation sites due to insufficient sequencing depth, while TGS identified all 11 sites and assigned the A1 phenotype. Concordance rates between serology and genotyping were: Duffy 96.55% (140/145), ABO 96.43% (513/532), Kidd 94.17% (97/103), MNS 85.07% (57/67), and Lewis 68.18% (75/110). In the MNS system, 7 of 8 samples serologically typed as M+N+ but genotyped as M-N+ carried GYPB variants. Lewis discrepancies predominantly featured genotype Le (a-b+) with serological phenotypes of Le (a+b-), Le (a+b+), or Le (a-b-). The NGS platform completed sequencing of 192 samples within 2 weeks. Conclusion: The tiered genotyping strategy combining NGS, TGS, and serology enables multi-system blood group identification in large-scale blood donor populations. TGS provides complementary value in phasing ambiguous ABO samples and detecting variants in regions with insufficient sequencing depth. This study provides a technical framework and baseline frequency data for the expansion of a local rare blood type repository. However, standardization and cost-effectiveness of this strategy require further validation with expanded sample sizes, and serological confirmation should be retained for secretion status-related systems such as Lewis.

OBJECTIVE To investigate the pharmacokinetics and bioavailability of naproxen choline ionic liquid in rats after intragastric administration. METHODS Naproxen choline ionic liquid was given to rats by intragastric administration. Blood samples were collected at different time points after administration. The blood samples were precipitated by methanol and then centrifuged, and then an Extend-C18 column(4.6 mm×250 mm, 5 μm) was used. Methanol(A)-0.3% phosphoric acid aqueous solution(B) (74:26) was used as the mobile phase, the flow rate was 0.8 mL·min-1, and the detection wavelength was 230 nm. Indomethacin and naproxen were used as internal standard and tested object in determination of naproxen choline ionic liquid in rat plasma. Pharmacokinetic parameters were calculated using DAS 2.0 software. RESULTS After intragastric administration of naproxen suspension to rats, its t1/2α was 5.12 h, t1/2β was 10.13 h, Tmaxwas 2 h, Cmax was 112.92 mg·L-1, and AUC(0-t) was 1 091.01 mg·L-1·h. After intragastric administration of naproxen choline, its t1/2α was 5.64 h, t1/2β was 69.32 h, Tmax was 1 h, Cmax was 135.97 mg·L-1, AUC(0-t) was 1 305.79 mg·L-1·h, and the relative bioavailability was 119.686%. CONCLUSION After intragastric administration of naproxen choline to rats, the peak concentration and bioavailability of the naproxen in vivo are higher than those of the common suspension, and the peak time is earlier.

Objective To investigate the difference of MT4 G-201A of Metallothionein (metallotionein, MT) family gene locus single nucleotide polymorphisms (SNPs) in type 2 diabetes and essential hypertension. Mothods We selected 324 cases of patients with type 2 diabetes, 301 cases of patients with essential hypertension, 301 case of normal physical examination population. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis technology was used to detect MT4 G-201A polymorphism. Results (1) The genotype GG, GA and AA frequency of MT4 G-201A in 324 cases of patients with type 2 diabetes,301 cases of patients with essential hypertension, 301 case of normal physical examination population were 39.5%, 46.6%, 13.9%;42.2%, 48.5%, 9.3%;and 42.5%, 51.5%, 6.0%, respectively. The genotypes in all the distribution were in Hardy-Weinberg equilibrium (P>0.05) . (2) MT4 G-201A polymorphism:Three groups of genotype and allele frequency distribution (G and A) had differences (P<0.05) ; there was no difference between essential hypertension group and normal group (P>0.05);there was a difference between type 2 diabetes group and the normal group (P<0.05);there was no significant difference in genotype and allele frequency distribution (P>0.05) . (3) MT4 G-201A polymorphism in Men:Three groups of genotype and allele frequency distribution had differences (P<0.05) . In essential hypertension and type 2 diabetes group, there was no significant difference in genotype and allele frequency distribution (P>0.05) .MT4 G-201A polymorphism between women:Three groups of genotype and allele frequency distribution had differences (P<0.05), there was a difference between type 2 diabetes and normal group genotypes (P<0.05) . Conclusions (1) There is correlation of MT gene G-210A polymorphism and type 2 diabetes, no association with essential hypertension. (2) There is a difference of MT gene G-210A polymorphism in type 2 diabetes in women, no difference in men.

Objective To study the analgesic effect of Eupatorium odoratum L and its mechanism .Methods Mice were random‐ly divided into 6 groups .Swinging tail method in spinal cord destroyed mouse ,hot‐plate and twisted body method in naloxone blocked mouse ,and formalin‐induce inflammatory pain method were done to investigate the analgesic effect of Eupatorium odoratum L .Each pain index under each experiment condition was determined after intra gastric administration of each group of mice .The concentration of NO and PGE2 in serum and brain were measured to study its mechanisms .Results High‐dose of Eupatorium odor‐atum L significantly enhanced pain threshold in either spinal cord destroyed or naloxone blocked mouse with pain induced by hot and acetic acid(P<0 .05) .High and middle dosage groups of Eupatorium odoratum L markedly reduced the first‐and second‐phase re‐sponses of formalin‐induced inflammatory pain mouse(P< 0 .05) ,and decreased the writhing times and licking rear feet times caused by twisted body method(P<0 .05) .The level of NO and PGE2 in serum and brain of mouse induced by formalin were both decreased(P<0 .05) .Conclusion Eupatorium odoratum L has notable analgesic effect ,which may be related to reduce the level of NO and PGE2 in central and peripheral site .