Objective: To characterize the serological profile, RHD genetic spectrum, and their frequencies among pregnant women preliminary screened as RhD-negative and weak positive in Qingdao and surrounding areas, and correlate these findings with unexpected antibody detection results, thereby providing testing recommendations and suggestions for such individuals. Methods: Blood samples of pregnant women who were initially identified as RhD negative and weak positive in hospitals in Qingdao and surrounding areas over the past five years were collected. Different cloned IgG anti-D antibodies were used for RhD negative confirmation experiments. RHD genotyping was performed by combining PCR-SSP and Sanger sequencing. Unexpected antibody screening and identification were carried out using test tube method and microcolumn gel card. The immunologic status of newborns delivered by anti-D pregnant women was also tracked. Results: A total of 602 blood samples were collected from pregnant women initially identified as RhD-negative and weak positive. Among them, 569 (94.5%) were confirmed as RhD-negative in the RhD confirmation test, and 33 (5.5%) were D variant phenotype. Except for 4 cases where no definite mutations were found, gene analysis revealed 474 (78.7%) D-negative cases with 5 genotypes (RHD 01N.01, RHD 01N.03, RHD 01N.16, RHD 01N.05, and 1 new allele), 90 (15.0%) Del cases with 2 genotypes (RHD 01EL.01, and RHD 01EL.18), 23 (3.8%) weak D cases with 2 genotypes (RHD 15 and 1 new allele), and 11 (1.8%) partial D cases with 2 genotypes (RHD 06.03.01 and RHD 05.04). Anti-D and complex antibodies containing anti-D were detected in 96 RhD-negative and partial D pregnant women (15.9%). After injection of anti-D immunoglobulin, One O-type RhD-negative pregnant woman delivered a newborn with hyperbilirubinemia. The newborn was typed to be B Del, and anti-D was detected in both serum and eluate. Conclusion: The serological profiles, RHD gene types and frequencies among RhD negative pregnant women in Qingdao and surrounding areas are basically consistent with domestic published data. Pregnancy can stimulate anti-D production in D-negative and partial D individuals. However, anti-D antibody has not been detected in Del type pregnant women. Since anti-D immunoglobulin can binds to Del type red blood cells, its administration is not recommended for Del type pregnant women.

Amblyopia is a common visual development disorder and is the main cause of monocular vision impairment in children and adults. Photobiomodulation(PBM), a non-invasive treatment method, has gradually gained attention in the field of ophthalmology. This paper begins with the macroscopic manifestation of light on the animal model of amblyopia. Additionally, it discusses the pathological changes of the amblyopic retina and the human eye's central nervous system, as well as the influence and mechanism of PBM on the visual perception and processing system and its chemical effect on the visual system through dopamine and melatonin. It examines its mechanism of action, current clinical application status, and future development direction in order to provide new ideas and theoretical foundation for amblyopia treatment.

Dry eye disease (DED) is a prevalent and intractable ocular disease induced by a variety of causes. Elevated sphingomyelin (SM) levels and pro-inflammatory cytokines were detected on the ocular surface of DED patients, particularly in the meibomian glands. Sphingomyelin synthase 2 (SMS2), one of the proteins involved in SM synthesis, would light a novel way of developing a DED therapy strategy. Herein, we report the design and optimization of a series of novel thiophene carboxamide derivatives to afford 14l with an improved highly potent inhibitory activity on SM synthesis (IC_{50, SMS2} = 28 nmol/L). Moreover, 14l exhibited a notable protective effect of anti-inflammation and anti-apoptosis on human corneal epithelial cells (HCEC) under TNF-α-hyperosmotic stress conditions in vitro, with an acceptable ocular specific distribution (corneas and meibomian glands) and pharmacokinetics (PK) profiles (t _{1/2, cornea} = 1.11 h; t _{1/2, meibomian glands} = 4.32 h) in rats. Furthermore, 14l alleviated the dry eye symptoms including corneal fluorescein staining scores and tear secretion in a dose-dependent manner in mice. Mechanically, 14l reduced the mRNA expression of Tnf-α, Il-1β and Mmp-9 in corneas, as well as the proportion of very long chain SM in meibomian glands. Our findings provide a new strategy for DED therapy based on selective SMS2 inhibitors.

Purpose: This study identified risk factors for neurogenic lower urinary tract dysfunction (NLUTD) in patients with acute ischemic stroke (AIS) through multidimensional analysis of the medical records of patients, aiming to reduce the incidence of NLUTD, improve prognosis, and facilitate rehabilitation. Materials and Methods: In this case-control study, patients with AIS were recruited from two tertiary general hospitals in Shenzhen, China, from March 2021 to October 2023. Patients were divided into NLUTD and non-NLUTD groups based on the presence and absence of NLUTD, respectively. Comparative analysis was performed using the Mann–Whitney U and chi-square tests, with significant variables being included in logistic regression analysis. Results: Of the 652 participants enrolled in this study, 119 participants (18.3%) developed NLUTD. Bivariate analysis showed that 39 of 54 screened factors exhibited a significant correlation (p<0.05) with the incidence of NLUTD after AIS. Significant variables identified through logistic regression analysis included Glasgow coma scale (GCS) and National Institutes of Health Stroke Scale (NIHSS) scores, anemia, aphasia, pneumonia, brainstem involvement, multiple lesions, urine clarity (CLA), random venous blood glucose (GLU) and hemoglobin (HGB) levels, and white blood cell (WBC) count. Conclusions A total of 11 risk factors for NLUTD were identified in this study. This finding provides valuable guidance for reducing the incidence of NLUTD after AIS and improving the quality of life of patients.

Alcoholic liver disease (ALD) is a growing global health concern, and its early pathogenesis includes steatosis and steatohepatitis. Inhibiting lipid accumulation and inflammation is a crucial step in relieving ALD. Evidence shows that puerarin (Pue), an isoflavone isolated from Pueraria lobata, exerts cardio-protective, neuroprotective, anti-inflammatory, antioxidant activities. However, the therapeutic potential of Pue on ALD remains unknown. In the study, both the NIAAA model and ethanol (EtOH)-induced AML-12 cell were used to explore the protective effect of Pue on alcoholic liver injury in vivo and in vitro and related mechanism. The results showed that Pue (100 mg·kg^-1) attenuated EtOH-induced liver injury and inhibited the levels of SREBP-1c, TNF-α, IL-6 and IL-1β, compared with silymarin (Sil, 100 mg·kg^-1). In vitro results were consistent within vivo results. Mechanistically, Pue might suppress liver lipid accumulation and inflammation by regulating MMP8. In conclusion, Pue might be a promising clinical candidate for ALD treatment.

Five new saponins, including three steroid saponins, paristenoids A-C (1-3), and two triterpenoid saponins, paristenoids D-E (4-5), along with four known ones (6-9) were isolated from the rhizomes of Paris polyphylla var. stenophylla. The structures of the isolated compounds were identified mainly by detailed spectroscopic analysis, including extensive 1D and 2D NMR, MS, as well as chemical methods. Compound 3 is a new cyclocholestanol-type steroidal saponin with a rare 6/6/6/5/5 fused-rings cholestanol skeleton, and this skeleton has been first found from the genus Paris. The cytotoxicities of the isolated compounds against three human three glioma cell lines (U87MG, U251MG and SHG44) were evaluated, and compound 7 displayed certain inhibitory effect with IC₅₀ values of 15.22 ± 1.73, 18.87 ± 1.81 and 17.64 ± 1.69 μmol·L^-1, respectively.
Humans ; Rhizome/chemistry* ; Steroids/chemistry* ; Liliaceae/chemistry* ; Saponins/chemistry* ; Triterpenes/analysis*

Objective: To construct a mouse derived pcDNA3. 1 ⁃3 × Flag⁃c⁃NUP85 expression plasmid and observe its effect on expression of inflammation factors in LPS⁃induced RAW264. 7 cells , as well as on the proliferation and apoptosis of RAW264. 7 cells. Methods: The NUP85 gene was amplified by PCR to construct pcDNA3. 1 ⁃3 × Flag-c⁃NUP85 eukaryotic expression plasmid. The pcDNA3. 1 ⁃3 × Flag⁃c vector was divided with enzymes. The purified PCR product was ligated with the vector, and the ligated product was transformed into bacterial competent cells. After identification by enzyme digestion , sequencing and analysis were performed. Then , it was transfected into RAW264. 7 cells , and the blank plasmid without NUP85 gene was set as the control group. The effect on cell proliferation and apoptosis were detected by CCK⁃8 assay and flow cytometry , and the expression of inflammatory cytokines such as tumor necrosis factor⁃α (TNF⁃α ) and interleukin⁃6 (IL⁃6) in LPS⁃induced RAW264. 7 cells was detected by Western blot and ELISA. Results: Enzyme digestion identification and Western blot results showed that pcDNA3. 1 ⁃3 × Flag⁃c⁃NUP85 eukaryotic expression plasmid was successfully constructed and expressed. The results of CCK⁃8 assay showed that the cell survival rate of NUP85 overexpression group was significantly lower than that of control group after 24 h[(0. 55 ± 0. 03) vs (0. 67 ± 0. 05) , F = 30. 98 , P < 0. 05 ] . The results of flow cytometry showed that the cell apoptosis rate of NUP85 overexpression group was higher than that of control group[( 15. 78 ±1. 05)% vs ( 13. 40 ± 0. 47)% , F = 75. 38 , P < 0. 05] . The results of Western blot and ELISA showed that after transfection of pcDNA3. 1 ⁃3 × Flag⁃c⁃NUP85 , the expression of TNF⁃α and IL⁃6 in RAW264. 7 cells were higher than those in the control group ,with statistical significance (P < 0. 05) . Conclusion NUP85 can inhibit the proliferation and promote apoptosis in LPS⁃stimulated RAW264. 7 cells , and NUP85 can promote the expression of inflammatory cytokines IL⁃6 and TNF⁃α in LPS⁃stimulated RAW264. 7 cells.

Objective: To investigate the proteomic characteristics of overweight/obesity and related abnormal glucose and lipid metabolism caused by phlegm-dampness retention to identify related biomarkers. Methods: Seventy-one subjects were enrolled in the study. We assessed blood glucose, blood lipids, body mass index (BMI), and phlegm-dampness pattern, which was confirmed by a traditional Chinese med-icine clinician. Of the participants, we included healthy participants with normal weight (NW, n =23), overweight/obese participants with normal metabolism (ONM, n = 19), overweight/obese participants with pre-diabetes (OPD, n = 12), and overweight/obese participants with marginally-elevated blood lipids (OML, n = 17). Among them, the ONM, OPD, and OML groups were diagnosed with phlegm-dampness pattern. The data-independent acquisition (DIA) method was first used to analyze the plasma protein expression of each group, and the relevant differential proteins of each group were screened. The co-expressed proteins were evaluated by Venn analysis. The pathway analyses of the differential proteins were analyzed using Ingenuity Pathway Analysis (IPA) software. Parallel reaction monitoring (PRM) was used to verify the differential and common proteins in each group. Results: After comparing ONM, OPD, and OML groups with NW group, we identified the differentially expressed proteins (DEPs). Next, we determined the DEPs among OPD, OML, and ONM groups. Using Venn analysis of the DEPs in each group, 24 co-expressed proteins were screened. Two co-expressed proteins were verified by PRM. IPA analysis showed that pathways including LXR/RXR activation, acute phase response signaling, and FXR/RXR activation were common to all three groups of phlegm-damp overweight/obesity participants. However, the activation or inhibition of these pathways was different among the three groups. Conclusion: Participants with overweight/obesity have similar proteomic characteristics, though each type shows specific proteomic characteristics. Two co-expressed proteins, VTN and ORM1, are potential biomarkers for glucose and lipid metabolism diseases with overweight/obesity caused by phlegm-dampness retention.

OBJECTIVE: To observe the therapeutic effect of different doses of dihydroartemisinin (DHA) on atopic dermatitis (AD) in mice and explore the mechanism. METHODS: Forty-two C57BL/6 mice were randomly divided into 7 groups ( RESULTS: Treatment with 25, 75, and 125 mg/kg DHA and dexamethasone all alleviated AD symptoms of mice, reduced the severity scores of skin lesions, and ameliorated pathological changes of the skin tissue. DHA at 125 mg/kg produced the most obvious therapeutic effect and significantly alleviated mast cell infiltration in the lesions as compared with the other treatment groups ( CONCLUSIONS DHA is effective for the treatment of AD in mice with an optimal dose of 125 mg/kg. The therapeutic effect of DHA is achieved probably through regulation of local immunity by inhibiting mast cell infiltration in the lesions.
Animals ; Anti-Inflammatory Agents/therapeutic use* ; Artemisinins ; Cytokines ; Dermatitis, Atopic/drug therapy* ; Immunoglobulin E ; Mast Cells ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Skin

OBJECTIVE To construct eukaryotic expression vectors for human ABO and subgroup genes A101, B101, CisAB01, Ael05, B(A)04 and Bw03, and validate their expression in vitro. METHODS Total RNA was isolated from individuals with the A101 and B101 subgroups. cDNA of A101 and B101 was synthesized by reverse transcription and amplified with specific primers. Subgroup genes CisAB01, Ael05, B(A)04 and Bw03 were then amplified with PCR for site-directed mutagenesis. Fragments of the ABO genes were directionally linked to pcDNA3.1 positive-eukaryotie expression vectors. After antibiotic screening, the sequences were analyzed. The vectors were transfected into Hela cells, and the expression of target proteins was detected by Western blotting and immunofluorescence assay. RESULTS Sanger sequencing has confirmed that pDNA3.1-A101, pDNA3.1-CisAB01, pDNA3.1-Ael05, pDNA3.1-B101, pDNA3.1- B(A)04, pDNA3.1-Bw03 positive-eukaryotic expression vector were successfully constructed. The results of Western blotting and immunofluorescence revealed clear presence of the expressed proteins. CONCLUSION Eukaryotic expression vectors for ABO subgroup genes were successfully constructed and worked well in Hela cells in vitro, which can facilitate further study of the ABO blood group proteins.