[Objective] To analyze the serological characteristics and molecular biology results for a P^k phenotype. [Methods] One patient with P^k phenotype upon unexpected antibodies at Jining Blood Center in July 2022 was selected as the study subject. The blood groups and unexpected antibodies of the proband and his second son were identified using serological methods. The sequences of 3-β-N-acetylgalactosaminyltransferase gene (B3GALNT1) and the coding region of α-1,4-galactosyltransferase gene (A4GALT) were amplified and analyzed by PCR direct sequencing, and haploid sequence analysis was carried out on the variant sites of the B3GALNT1 gene. PROVEAN, SIFT, PolyPhen2 and Mutation Taster were used to analyze the effect of mutations on the protein. [Results] Serological test results suggested that the proband was a P2^k type, including anti-P antibody in his serum, and his son was the P2 phenotype. Sequencing analysis revealed that the proband had a compound heterozygous variants of the B3GALNT1 in c. 239T>A (p. Leu80Gln) and c. 433C>T (p. Arg145Ter). Further haploid analysis showed that the c. 239T>A had occurred in one haploid while c. 433C>T was present in the other haploid, and his son carried a heterozygous variant of c. 433C>T (p. Arg145Ter); the proband and his son all have homozygous mutation variants of the A4GALT in c. 903C>G. Bioinformatic analysis also predicted that the mutations were harmful to the protein function. [Conclusion] The compound heterozygous variants c. 239T>A (p. Leu80Gln) and c. 433C>T (p. Arg145Ter) in the B3GALNT1 gene may contribute to the P^k phenotype, of which the c. 239T>A variant has not been reported.

OBJECTIVES: To propose a hypothesis that aloe-emodin may inhibit scar tissue fibrosis through thrombospondin-1(THBS1)-PI3K-Akt pathway. METHODS: By cultivating fibroblasts derived from scar tissue after cleft palate surgery in humans, aloe emodin of different concentrations (10, 20, 30, 40 and 50 μmol/L) was added to the cells which activity was detected. At the same time, transcriptome sequencing was performed on scar tissue and cells, and bioinformatics methods were used to explore potential targets and signaling pathways of scar tissue fibrosis. RESULTS: Aloe-emodin had a concentration dependent inhibitory effect on fibroblast proliferation,with the 40 μmol/L concentration group showing the most significant effect. The results of tissue and cell sequencing indicated that differentially expressed genes were significantly enriched in extracellular matrix-receptor interaction pathway, and shared a common differential gene which was THBS1. The ORA analysis results indicated that differentially expressed genes, including THBS1, were significantly enriched in the PI3K-Akt signaling pathway. CONCLUSIONS Aloe emodin may inhibit the PI3K-Akt pathway by downregulating THBS1, thereby reducing the proliferation activity of fibroblasts derived from postoperative palatal scar tissue.
Thrombospondin 1/genetics* ; Humans ; Signal Transduction/drug effects* ; Fibroblasts/cytology* ; Proto-Oncogene Proteins c-akt/metabolism* ; Fibrosis ; Phosphatidylinositol 3-Kinases/metabolism* ; Cicatrix/metabolism* ; Cell Proliferation/drug effects* ; Anthraquinones/pharmacology* ; Cells, Cultured

Objective To evaluate the forensic application value of used dental floss as a source of bio-logical evidence for individual identification by analyzing the effects of dental floss sample collection methods,DNA extraction methods,preservation conditions,and sampling sites on the success rate of STR typing.Methods Dental floss samples were collected using three techniques:direct cutting,cotton swab wiping,and flocked swab wiping,respectively.DNA was extracted respectively by the Chelex,spin column-based and magnetic bead-based methods.DNA quantification and STR typing were per-formed using the Qubit kit and FGI HumDNA Typing kit(Platinum),respectively.Storage environ-ments(temperature and humidity,ultraviolet radiation)and sampling locations(the floss part,the handle part)on DNA quantity and STR typing were evaluated.Results Through conducting a statisti-cal analysis of three key indicators of average DNA mass concentration,STR locus detection rate,and typing accuracy rate,the direct cutting method demonstrated the highest efficacy,followed by cotton swab wiping mothed,and the flocked swab wiping method had the lowest efficacy.Direct cutting yielded an average DNA mass concentration greater than(4.94±1.87)ng/μL,with STR locus detection and accuracy rates of 100%.Bead-based DNA extraction method produced superior DNA concentration and quality compared to spin column-based and Chelex methods,regardless of whether the sampling technique used.Preservation conditions had a significant impact on the DNA analysis of samples.Par-ticularly,the STR typing accuracy of samples preserved at 55℃/50%RH for 35 days dropped to(81.82±12.31)%,and that of samples exposed to ultraviolet radiation for 12 h dropped to(55.46±34.31)%.DNA concentration from the handle part of dental floss was extremely low,with an STR typing accuracy of only(30.91±27.35)%.Conclusion Using cotton swabs to wipe or directly cutting the thread of dental floss samples,and combining this approach with the magnetic bead method for DNA extraction,can best guarantee the concentration and quality of DNA.In addition,samples should be stored in low-temperature,low-humidity environment,protected from light and ultraviolet radiation.

AIM To explore the clinical effects of Tongfu Jiangzhuo Formula on ICU delirium patients with Internal Accumulation of Turbid Toxins.METHODS Based on Turbid Toxins theory,two hundred and ten patients were randomly assigned into control group(105 cases)for intervention of conventional treatment,and observation group(105 cases)for intervention of both Tongfu Jiangzhuo Formula and conventional treatment.The changes in clinical effects,delirium scores(CAM-ICU,ICDSC),TCM syndrome score,inflammatory factors(IL-1β,IL-6,IL-8),Raf-1,MEK-2,ERK-1 mRNA and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05).After the treatment,the two groups displayed decreased delirium scores,TCM syndrome score,inflammatory factors and Raf-1,MEK-2,ERK-1 mRNA(P＜0.05),especially for the observation group(P＜0.05).No significant difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with ICU delirium due to Internal Accumulation of Turbid Toxins,Tongfu Jiangzhuo Formula can safely and effectively improve symptoms,and reduce inflammation levels and Raf-1,MEK-2,ERK-1 mRNA expressions.

AIM To explore the clinical effects of Tongfu Jiangzhuo Formula on ICU delirium patients with Internal Accumulation of Turbid Toxins.METHODS Based on Turbid Toxins theory,two hundred and ten patients were randomly assigned into control group(105 cases)for intervention of conventional treatment,and observation group(105 cases)for intervention of both Tongfu Jiangzhuo Formula and conventional treatment.The changes in clinical effects,delirium scores(CAM-ICU,ICDSC),TCM syndrome score,inflammatory factors(IL-1β,IL-6,IL-8),Raf-1,MEK-2,ERK-1 mRNA and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05).After the treatment,the two groups displayed decreased delirium scores,TCM syndrome score,inflammatory factors and Raf-1,MEK-2,ERK-1 mRNA(P＜0.05),especially for the observation group(P＜0.05).No significant difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with ICU delirium due to Internal Accumulation of Turbid Toxins,Tongfu Jiangzhuo Formula can safely and effectively improve symptoms,and reduce inflammation levels and Raf-1,MEK-2,ERK-1 mRNA expressions.

AIM: To compare the outcome of C3F8 versus silicone oil tamponade after pars plana vitrectomy(PPV)and inverted internal limiting membrane(ILM)for the treatment of highly myopic macular hole retinal detachment(MHRD).METHODS: Retrospective clinical study. Totally 45 patients(45 eyes)with highly myopic MHRD who visited our hospital between January 2019 and August 2022 were selected as the research subjects. The patients were divided into two groups according to different intraocular tamponade agents: C3F8(22 eyes)and silicone oil(23 eyes)groups. All patients underwent conventional three-incision PPV, ILM was tamped, a venous blood clot was placed on the tamped ILM, and 15% C3F8 and silicone oil were used as tamponade, respectively. The best corrected visual acuity(BCVA), multifocal electroretinogram(mfERG), the closure of the macular hole, retinal reattachment and the complications were observed.RESULTS: The macular hole closure rate was 77% in the C3F8 group and 83% in the silicone oil group, respectively(P>0.05), and retinal reattachment rates were 95% and 96%, respectively(P>0.05). The visual acuity of the two groups significantly improved, which was 0.99±0.34 and 1.22±0.37, respectively, and the C3F8 group was better than that of the silicone oil group(t=-2.156, P=0.037). After operation, the response density of the first ring of P1 wave in the first order kernel in mfERG was 114.27±26.37 nV/deg2 for the C3F8 group and 98.08±24.36 nV/deg2 for the silicone oil group, and the response density of the second ring of P1 wave was 80.45±14.94 nV/deg2 for the C3F8 group and 67.73±15.33 nV/deg2 for the silicone oil group, all of which were significantly higher compared to pre-operation [the response density of the first ring of P1 wave: 58.13±13.96 nV/deg2 for the C3F8 group and 55.30±10.48 nV/deg2 for the silicone oil group, the response density of the second ring of P1 wave: 51.18±8.19 nV/deg2 for the C3F8 group and 47.43±11.97 nV/deg2 for the silicone oil group](all P<0.05). It was found that the response density of the first ring of P1 wave was lower in the silicone oil group than in the C3F8 group(P<0.05). There was no statistically significant difference in the incidence of complications between the two groups(P>0.05).CONCLUSION: Silicone oil tamponade or C3F8 tamponade after PPV combined with ILM can both promote retinal reattachment and macular hole closure in patients with MHRD, and the C3F8 tamponade was superior to silicone oil in visual function recovery.

Objective To investigate the mechanism of RNF7 for regulating myeloid-derived suppressor cells(MDSCs)in non-small cell lung cancer(NSCLC).Methods TIMER2.0 database and immunohistochemistry were used to analyze RNF7 expression level and its correlation with immune cell infiltration in non-small cell lung cancer.The impact of RNF7 expression levels on prognosis of lung cancer patients was analyzed using Kaplan-Meier survival analysis.CMT-167 cells with RNF7 overexpression or knockdown were inoculated subcutaneously in C57BL/6 mice,and the mice in RNF7 knockdown group were treated with anti-PD-1 or IgG isotype control 7 days after the inoculation.The tumor tissues were harvested after 30 days for tumor volume measurement,detection of S100A8+A9 and Gr-1 expressions with immunohistochemistry,and analysis of MDSC infiltration.Gene set enrichment analysis(GSEA)was performed to identify the potential pathways regulated by RNF7 in NSCLC.Western blotting and luciferase assays were used to assess the impact of RNF7 on the NF-κB signaling pathway.ELISA and RT-qPCR were used to measure chemokine(C-X-C motif)ligand 1(CXCL1)expression.Results RNF7 expression was significantly upregulated in NSCLC,and high RNF7 expression levels were associated with poor prognosis of the patients(P<0.001).TIMER2.0 analysis revealed a positive correlation between RNF7 expression and MDSC infiltration(P<0.001).GSEA suggested that RNF7 was enriched in the NF-κB signaling pathway.In NSCLC cells,RNF7 knockdown significantly inhibited NF-κB activation and reduced CXCL1 expression.In the tumor-bearing mice,RNF7 overexpression significantly increased MDSC infiltration in the tumor tissue,and RNF7 knockdown obviously reduced MDSC infiltration and enhanced the efficacy of anti-PD-1 therapy.Conclusion High expression of RNF7 in NSCLC cells promotes CXCL1 expression by activating the NF-κB signaling pathway,thus leading to the chemotactic recruitment of MDSCs,which contributes to tumor resistance to anti-PD-1 therapy.

Objective To analyzes the influence of Artificial Intelligence(AI)technology on physician workload based on the panel data of 31 provinces in China from 2012 to 2021.Methods Using MATLAB R2022b software to estimate the kernel density of physician workload and analyze the changes of physician workload in the past 10 years.Select fixed effects model and perform panel regression analysis between AI techniques and physician workload based on Stata MP 17.0 software.Results At present,there is a serious regional multi-level differentiation in physician workload.AI-assisted diagnosis,AI-assisted treatment and AI-assisted management have significantly reduced the physician workload.However,there is regional difference in the negative influence of AI technology on physician workload,with the most significant influence in eastern China and less significant in western China.Both aging and personal health investment negatively moderate the relationship between AI-assisted diagnosis technology and physician workload.Conclusion Great attention should be paid to AI,especially AI-assisted diagnosis technology,because it plays an important role in reducing the physician workload and alleviating the pressure caused by insufficient medical resources and uneven distribution in the context of aging.

Objective To investigate the mechanism of RNF7 for regulating myeloid-derived suppressor cells(MDSCs)in non-small cell lung cancer(NSCLC).Methods TIMER2.0 database and immunohistochemistry were used to analyze RNF7 expression level and its correlation with immune cell infiltration in non-small cell lung cancer.The impact of RNF7 expression levels on prognosis of lung cancer patients was analyzed using Kaplan-Meier survival analysis.CMT-167 cells with RNF7 overexpression or knockdown were inoculated subcutaneously in C57BL/6 mice,and the mice in RNF7 knockdown group were treated with anti-PD-1 or IgG isotype control 7 days after the inoculation.The tumor tissues were harvested after 30 days for tumor volume measurement,detection of S100A8+A9 and Gr-1 expressions with immunohistochemistry,and analysis of MDSC infiltration.Gene set enrichment analysis(GSEA)was performed to identify the potential pathways regulated by RNF7 in NSCLC.Western blotting and luciferase assays were used to assess the impact of RNF7 on the NF-κB signaling pathway.ELISA and RT-qPCR were used to measure chemokine(C-X-C motif)ligand 1(CXCL1)expression.Results RNF7 expression was significantly upregulated in NSCLC,and high RNF7 expression levels were associated with poor prognosis of the patients(P<0.001).TIMER2.0 analysis revealed a positive correlation between RNF7 expression and MDSC infiltration(P<0.001).GSEA suggested that RNF7 was enriched in the NF-κB signaling pathway.In NSCLC cells,RNF7 knockdown significantly inhibited NF-κB activation and reduced CXCL1 expression.In the tumor-bearing mice,RNF7 overexpression significantly increased MDSC infiltration in the tumor tissue,and RNF7 knockdown obviously reduced MDSC infiltration and enhanced the efficacy of anti-PD-1 therapy.Conclusion High expression of RNF7 in NSCLC cells promotes CXCL1 expression by activating the NF-κB signaling pathway,thus leading to the chemotactic recruitment of MDSCs,which contributes to tumor resistance to anti-PD-1 therapy.