Perilla frutescens (L.) Britt. is a characteristic oil crop rich in polyunsaturated fatty acids, particularly α-linolenic acid, which has important development and utilization value. The Dof transcription factor is one of the plant-specific transcription factor families, which is widely involved in important biological processes such as plant growth, development, and metabolic regulation. In order to explore the key Dof transcription factors involved in the oil biosynthesis and systematically analyze their regulatory mechanisms of P. frutescens seeds, a total of 56 PfDof gene family members were identified from the genome and transcriptome data of P. frutescens and classified into four subfamilies according to sequence characteristics. All PfDofs contained highly conserved C₂-C₂ zinc finger domains, with gene duplication being the primary mechanism driving their evolution and expansion. Genes within the same subgroup exhibited similar gene structures and conserved motifs. The 56 PfDofs were predicted as unstable hydrophilic proteins, with α-helixes and random coils as their predominant structural components. The RNA-seq results revealed that 11 PfDofs exhibited differential expression during different developmental stages of P. frutescens seeds. RT-qPCR was performed to further validate the expression patterns of these 11 members across various tissue samples (root, stem, leaf, and flower) of P. frutescens and at different developmental stages of its seeds. The results showed that PfDof29 exhibited the highest expression level in seeds, which was consistent with the transcriptome data. Subcellular localization studies demonstrated that PfDof29 was localized to the nucleus and had a transcriptional activation activity. Overexpression of PfDof29 in Nicotiana tabacum resulted in a significant increase in total oil content of tobacco leaves, accompanied by reductions in starch and soluble sugar content, while the protein content remained unchanged. Additionally, the metabolic balance between saturated and unsaturated fatty acids in the transgenic tobacco leaves was altered, with a significant increase in α-linolenic acid content. The expression levels of the fatty acid desaturase genes NtFAD2, NtFAD3, and NtFAD8 were significantly upregulated. A yeast one-hybrid assay revealed that PfDof29 could directly bind to the promoter region of PfFAD8, thereby regulating its expression. This study provides an initial understanding of the regulatory mechanisms of PfDof transcription factors in the synthesis and accumulation of oil in P. frutescens. These findings offer new insights into the enhancement of oil content and quality of P. frutescens seeds.
Transcription Factors/physiology* ; Perilla frutescens/metabolism* ; Plant Proteins/metabolism* ; Gene Expression Regulation, Plant ; alpha-Linolenic Acid/biosynthesis* ; Lipids/biosynthesis* ; Seeds/genetics*

To establish a method for determining 26 inorganic elements in earthworm polypeptide and determine the elemental content in different batches of earthworm polypeptide, microwave digestion method was used to pre-treat the samples, and ICP-MS method was used to determine the content of 26 elements in different batches of earthworm polypeptide. The linear relationships of 26 elements were good in the range of 0-1 000 μg·L^-1, with R² greater than 0.999, precision RSD 0.21%-2.71%, repeatability RSD 0.19%-4.69%, stability RSD 0.11%-4.24%, and recovery rates of 82.41%-116.16%. The data was plotted using Origin 2022 software to characterize the distribution of elements content. SPSS 27.0 was used for principal component analysis, and SIMCA 14.1 software was used for OPLS-DA analysis. The results showed that among the 26 elements, the higher content of earthworm polypeptide was K, Na, and Ca, followed by Zn, Fe, Al, B and other trace elements, In, Sc, Co, Pb, Bi and other elements had little or no detectable content, and there were differences in the content of polypeptide in different batches. This study provides a theoretical basis for the production quality control, quality evaluation and drug efficacy application of earthworm polypeptide through the determination and analysis of the elements and content of earthworm polypeptide.

The density and composition of lymphocytes infiltrating colon tumors serve as predictive factors for the clinical outcome of colon cancer.Our previous studies highlighted the potent anti-cancer properties of the principal compounds found in Garcinia yunnanensis(YTE-17),attributing these effects to the regu-lation of multiple signaling pathways.However,knowledge regarding the mechanism and effect of YTE-17 in the prevention of colorectal cancer is limited.In this study,we conducted isobaric tags for relative and absolute quantification(iTRAQ)analysis on intestinal epithelial cells(IECs)exposed YTE-17,both in vitro and in vivo,revealing a significant inhibition of the Wnt family member 5a(Wnt5a)/c-Jun N-terminal kinase(JNK)signaling pathway.Subsequently,we elucidated the influence and mechanism of YTE-17 on the tumor microenvironment(TME),specifically focusing on macrophage-mediated T helper 17(Th17)cell induction in a colitis-associated cancer(CAC)model with Wnt5a deletion.Additionally,we performed the single-cell RNA sequencing(scRNA-seq)on the colonic tissue from the Wnt5a-deleted CAC model to characterize the composition,lineage,and functional status of immune mesenchymal cells during different stages of colorectal cancer(CRC)progression.Remarkably,our findings demon-strate a significant reduction in M2 macrophage polarization and Th17 cell phenotype upon treatment with YTE-17,leading to the restoration of regulatory T(Treg)/Th17 cell balance in azoxymethane(AOM)/dextran sodium sulfate(DSS)model.Furthermore,we also confirmed that YTE-17 effectively inhibited the glycolysis of Th17 cells in both direct and indirect co-culture systems with M2 macrophages.Notably,our study shed light on potential mechanisms linking the non-canonical Wnt5a/JNK signaling pathway and well-established canonical β-catenin oncogenic pathway in vivo.Specifically,we proposed that Wnt5a/JNK signaling activity in IECs promotes the development of cancer stem cells with β-catenin activity within the TME,involving macrophages and T cells.In summary,our study undergoes the po-tential of YTE-17 as a preventive strategy against CRC development by addressing the imbalance with the immune microenvironment,thereby mitigating the risk of malignancies.

Objective:To observe the effectiveness and safety of pars plana vitrectomy (PPV) combined with inner limiting membrane (ILM) removal and 41G microneedle subretinal injection of balanced salt solution (BSS) in the treatment of refractory macular hole.Methods:A prospective clinical study. From January to June 2023, 20 cases (20 eyes) of refractory macular hole patients diagnosed through examination at The Affiliated Eye Hospital of Nanchang University were included in the study. The basal diameter of the affected eye's basal diameter (BD) was >1 000 μm. Macular hole index (MHI) was <0.5. The affected eye received treatment with 23G PPV combined with ILM removal and 41G microneedle subretinal injection of BSS. Best corrected visual acuity (BCVA), microperimetry, and optical coherence tomography angiography (OCTA) were performed before and 1, 2, 3, and 6 months after surgery for the affected eye. BCVA examination was performed using standard logarithmic visual acuity chart, and convert it to logarithmic minimum resolution angle (logMAR) visual acuity for statistical purposes. MP-3 microperimetry was used for micro view examination, record the mean sensitivity (MS) of the retinal within a 12° range of the fovea. OCTA was used to measure the area of the avascular zone of the macula (FAZ), perimeter of the FAZ (PERIM), retinal vascular length density (VLD), and vascular perfusion density (VPD). The changes in BCVA, MS, FAZ area, PERIM, VLD, VPD before and after surgery were compared and analyzed. After the same time, the closure of macular hole and the occurrence of complications after surgery were observed. Single factor analysis of variance was used to compare the observation indicators at different times before and after surgery. The correlation between various observation indicators and preoperative minimum diameter (MD), BD, and hiatus height at 6 months after surgery were analyzed using Pearson correlation analysis.Results:Among the 20 cases with 20 eyes, there were 2 males with 2 eyes and 18 females with 18 eyes. Age was (61.45±8.56) years old. The logMAR BCVA, MS, FAZ area, PERIM, VLD, and VPD of the affected eye were 1.46±0.21, (16.20±5.81) dB、(0.40±0.17) mm 2, (2.89±0.99) mm, (6.23±3.59) mm -1, (0.17±0.10)%, respectively. Six months after surgery, out of 20 eyes, macular hole closure and incomplete closure were 18 (90.0%, 18/20) and 2 (10.0%, 2/20) eyes, respectively. The logMAR BCVA, MS, FAZ area, PERIM, VLD, and VPD were 0.80±0.20, (22.20±4.60) dB, (0.18±0.10) mm 2, (1.83±0.80) mm, (9.54±2.88) mm -1, (0.31±0.14)%. Compared with before surgery, the differences were statistically significant ( P<0.05). The correlation analysis results showed a positive correlation ( P<0.05) between preoperative BD and postoperative 6-month PERIM and VPD. There was a negative correlation between preoperative MD and postoperative VLD at 6 months ( P<0.05). There was a negative correlation between preoperative MHI and logMAR BCVA and VPD at 6 months after surgery ( P<0.05). No complications such as elevated or decreased intraocular pressure, damage to retinal pigment epithelium, retinal hemorrhage, endophthalmitis, or retinal detachment occurred after surgery in all affected eyes. Conclusion:Minimally invasive PPV combined with ILM removal and 41G microneedle subretinal injection of BSS can effectively improve the closure rate of refractory macular hole patients in the short term, improve vision, and have good safety.

We wished to establish an expert consensus on late stage of critical care (CC) management. The panel comprised 13 experts in CC medicine. Each statement was assessed based on the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) principle. Then, the Delphi method was adopted by 17 experts to reassess the following 28 statements. (1) ESCAPE has evolved from a strategy of delirium management to a strategy of late stage of CC management. (2) The new version of ESCAPE is a strategy for optimizing treatment and comprehensive care of critically ill patients (CIPs) after the rescue period, including early mobilization, early rehabilitation, nutritional support, sleep management, mental assessment, cognitive-function training, emotional support, and optimizing sedation and analgesia. (3) Disease assessment to determine the starting point of early mobilization, early rehabilitation, and early enteral nutrition. (4) Early mobilization has synergistic effects upon the recovery of organ function. (5) Early functional exercise and rehabilitation are important means to promote CIP recovery, and gives them a sense of future prospects. (6) Timely start of enteral nutrition is conducive to early mobilization and early rehabilitation. (7) The spontaneous breathing test should be started as soon as possible, and a weaning plan should be selected step-by-step. (8) The waking process of CIPs should be realized in a planned and purposeful way. (9) Establishment of a sleep-wake rhythm is the key to sleep management in post-CC management. (10) The spontaneous awakening trial, spontaneous breathing trial, and sleep management should be carried out together. (11) The depth of sedation should be adjusted dynamically in the late stage of CC period. (12) Standardized sedation assessment is the premise of rational sedation. (13) Appropriate sedative drugs should be selected according to the objectives of sedation and drug characteristics. (14) A goal-directed minimization strategy for sedation should be implemented. (15) The principle of analgesia must be mastered first. (16) Subjective assessment is preferred for analgesia assessment. (17) Opioid-based analgesic strategies should be selected step-by-step according to the characteristics of different drugs. (18) There must be rational use of non-opioid analgesics and non-drug-based analgesic measures. (19) Pay attention to evaluation of the psychological status of CIPs. (20) Cognitive function in CIPs cannot be ignored. (21) Delirium management should be based on non-drug-based measures and rational use of drugs. (22) Reset treatment can be considered for severe delirium. (23) Psychological assessment should be conducted as early as possible to screen-out high-risk groups with post-traumatic stress disorder. (24) Emotional support, flexible visiting, and environment management are important components of humanistic management in the intensive care unit (ICU). (25) Emotional support from medical teams and families should be promoted through"ICU diaries"and other forms. (26) Environmental management should be carried out by enriching environmental content, limiting environmental interference, and optimizing the environmental atmosphere. (27) Reasonable promotion of flexible visitation should be done on the basis of prevention of nosocomial infection. (28) ESCAPE is an excellent project for late stage of CC management.
Humans ; Consensus ; Critical Care/methods* ; Intensive Care Units ; Pain/drug therapy* ; Analgesics/therapeutic use* ; Delirium/therapy* ; Critical Illness

Objective: To summarize the clinical characteristics and gene variants of 2 pedigrees of non-muscle myosin heavy chain 9 related diseases (MYH9-RD) in children. Methods: The basic information, clinical features, gene variants and laboratory tests of MYH9-RD patients from 2 pedigrees confirmed in the First Affiliated Hospital of Zhengzhou University in November 2021 and July 2022 were analyzed retrospectively. "Non-muscle myosin heavy chain 9 related disease" "MYH9" and "children" were used as key words to search at Pubmed database, CNKI and Wanfang database up to February 2023. The MYH9-RD gene variant spectrum and clinical data were analyzed and summarized. Results: Proband 1 (male, 11 years old) sought medical attention due to epistaxis, the eldest sister and second sister of proband 1 only showed excessive menstrual bleeding, the skin and mucous membrane of the their mother were prone to ecchymosis after bumping, the uncle of proband 1 had kidney damage, and the maternal grandmother and maternal great-grandmother of proband 1 had a history of cataracts. There were 7 cases of phenotypic abnormalities in this pedigree. High-throughput sequencing showed that the proband 1 MYH9 gene had c.279C>G (p.N93K) missense variant, and family verification analysis showed that the variant was inherited from the mother. A total of 4 patients including proband 1 and family members were diagnosed with MYH9-RD. The proband 2 (female, 1 year old) sought medical attention duo to fever and cough, and the father's physical examination revealed thrombocytopenia. There were 2 cases of phenotypic abnormalities in this pedigree. High-throughput sequencing showed that there was a c.4270G>A (p.D1424N) missense variant in the proband 2 MYH9 gene, and family verification analysis showed that the variant was inherited from the father. A total of 2 patients including proband 2 and his father were diagnosed with MYH9-RD. A total of 99 articles were retrieved, including 32 domestic literatures and 67 foreign literatures. The MYH9-RD cases totaled 149 pedigrees and 197 sporadic patients, including 2 pedigrees in our study. There were 101 cases with complete clinical data, including 62 sporadic cases and 39 pedigrees. There were 56 males and 45 females, with an average age of 6.9 years old. The main clinical manifestations were thrombocytopenia, skin ecchymosis, and epistaxis. Most patients didn't receive special treatment after diagnosis. Six English literatures related to MYH9-RD caused by c.279C>G mutation in MYH9 gene were retrieved. Italy reported the highest number of cases (3 cases). Twelve literatures related to MYH9-RD caused by c.4270G>A mutation in MYH9 gene were retrieved. China reported the highest number of cases (9 cases). Conclusions: The clinical manifestations of patients in the MYH9-RD pedigrees varied greatly. MYH9 gene c.279C>G and c.4270G>A mutations are the cause of MYH9-RD.
Infant ; Humans ; Female ; Male ; Child ; Myosin Heavy Chains/genetics* ; Ecchymosis ; Epistaxis ; Pedigree ; Retrospective Studies ; Muscular Diseases ; Thrombocytopenia ; Cytoskeletal Proteins

Most drugs need to interact with cell membrane to reach the biological target, so that membrane affinity assay is an important early screening step in drug discovery. However, at present, the traditional oil-water distribution method is still used, a new, simple and accurate method for membrane affinity assay is urgently needed. In this study, according to the colorimetric principle, a new assay model based on polydiacetylene vesicles was optimized through a series of experiments including different concentrations of vesicle solution, temperature, or pH reaction environment. On this basis, tetracaine hydrochloride, 2-methylimidazole and histamine were used as model drugs to measure the membrane affinity constants and verify the between-batch precision of the optimized assay model (relative standard deviation less than 5%). In addition, polydiacetylene vesicles were stable for up to 180 days, demonstrating the potential application of the assay model. This strategy is simple, stable, reliable, with high reproducibility, low cost and easy to promote, which provided a new tool and a new direction for the high-throughput assay of membrane affinity.

OBJECTIVE: To explore the effect of dichloromethane extraction phase of ethanol extract from stem of Patrinia scabiosaefolia Fisch.(DPSS) on proliferation and differentiation of K562 cells and its related mechanism. METHODS: MTT assay was used to detect the effects of DPSS at 0, 25, 50, 100 and 200 μg/ml on the proliferation of K562 cells at 24, 48 and 72 hours. Flow cytometry was used to analyze the changes of cell cycle and apoptosis at 24 and 48 hours. Wright-Giemsa staining was used to observe the morphological changes of K562 cells. The cell surface antigens CD33 and CD11b were detected by flow cytometry. RESULTS: The proliferation of K562 cells treated with different concentrations of DPSS was inhibited in a time-dose dependent manner (r=-0.96). Cell cycle analysis showed that with the increase of DPSS concentration, cells in G₂/M phase increased (r=0.88), and cells were blocked in G₂/M phase. Flow cytometry results showed that with the apoptosis rate of K562 cells was the highest when treated with 200 μg/ml DPSS for 48 h. Morphological observation showed that the K562 cell body increased, the amount of cytoplasm increased, the ratio of nucleus to cytoplasm decreased, and the nuclear chromatin was rough after DPSS treatment. Cell differentiation antigen, CD33 and CD11b, were positively expressed after treated with DPSS. CONCLUSION DPSS can induce apoptosis through cell cycle arrest, inhibit the proliferation of K562 cells, and induce K562 cells to differentiate into monocytes, which has a potential anti-leukemia effect.
Humans ; K562 Cells ; Patrinia ; Methylene Chloride/pharmacology* ; Apoptosis ; Cell Proliferation ; Cell Differentiation

OBJECTIVE: To investigate the clinical efficacy of femoral neck system(FNS) and three cannulated compression screws(CCS) in the treatment of unstable femoral neck fractures in young adults. METHODS: The clinical data of 52 young and middle-aged patients with unstable femoral neck fractures admitted from August 2018 to August 2021 were reviewed and analyzed. All patients were divided into two groups according to the internal fixation method, 25 cases were treated with FNS fixation, 27 cases were treated with closed reduction and 3 CCS inverted triangular distribution. The operation time, incision length, intraoperative bleeding, hospitalization expenses and fracture reduction quality of two groups were recorded and compared;The patients were followed up regularly after operation. The fracture healing time, complete weight-bearing time and postoperative complications(nonunion, femoral neck shortening, femoral head necrosis) of two groups were compared. The Harris score was used to evaluate hip function 6 months after surgery. RESULTS: The operation was successfully completed in both groups. The patients in FNS group had more bleeding, longer incision length and higher hospitalization cost than CCS group(P<0.01). There ware no significant difference in operation time and Garden index between two groups(P>0.05). Patients in both groups were followed up for 6 to 32 months.The fracture healing time in FNS group was less than that in CCS group, the time of complete weight bearing after surgery was earlier than that in CCS group, and the hip Harris score was higher than that in CCS group (P<0.01). There were no internal fixation fracture complications in two groups during follow-up. In the FNS group, there were 4 cases of avascular necrosis of the femoral head and 2 cases of femoral neck shortening, of which 3 cases underwent total hip replacement due to avascular necrosis of the femoral head. In the CCS group, there were 2 cases of nonunion, 9 cases of avascular necrosis of the femoral head, and 11 cases of femoral neck shortening, among which 5 cases underwent total hip replacement due to nonunion and avascular necrosis of the femoral head. CONCLUSION With simple operation, rotational stability and angular stability, FNS enables patients to start functional exercise as early as possible and reduces the incidence of postoperative complications of unstable femoral neck fracture. It is a new choice for the treatment of unstable femoral neck fracture in young adults.

Objective:To observe the efficacy and safety of ultra photon intense pulsed light combined with hydroquinone cream for the treatment of chloasma.Methods:From January 2020 to October 2022, a total of 30 patients with chloasma treated in the Department of Dermatology, Huazhong University of Science and Technology Union Shenzhen Hospital, ranged from 28 to 54 years old with an average age of 39 years, were randomly divided into three groups with 10 patients in each group: combined treatment group was treated with ultra photon intense pulsed light combined with hydroquinone cream; ultra photon intense pulsed light group was treated with ultra photon intense pulsed light alone; hydroquinone group was treated with hydroquinone cream alone. The efficacy and safety of the three groups were compared after four months of treatment.Results:After the treatment, 10 cases were effective in the combined treatment group, 5 cases were effective in the ultra photon intense pulsed light group and 2 cases were effective in the hydroquinone group. The total effective rate of the combined treatment group was significantly better than that of the ultra photon intense pulsed light group and the hydroquinone group, with statistical significance ( P<0.05). The average melasma area severity index score of the three groups after treatment was significantly decreased compared with that before treatment respectively ( P<0.05), and the average melasma area severity index score of the combined treatment group after treatment was significantly lower than that of hydroquinone group, with statistical significance ( P<0.05). No serious adverse reactions were observed in all the patients. Conclusions:Ultra photon intense pulsed light and hydroquinone cream can improve chloasma alone, but ultra photon intense pulsed light combined with hydroquinone cream has better therapeutic effect.