Objective To construct a 293T-CymR cell strain to express cysteine metabolism regulator(CymR) component by gene editing technology, and to regulate the target gene expression with CuO regulatory sequence promoter.Methods Based on clustered regularly interspaced short palindromic repeats(CRISPR)-Cas9 technology, the intracellular β factor receptor, C-C chemokine receptor(CCR5) gene, one of the safe harbors in HEK293T cell genome, was chosen as the target site for inserting the desired gene. The selection gene Pac(denoted as PuroR) encoding puromycin N-acetyltransferase was linked to the 3'end of the CymR gene via an internal ribosome entry site(IRES), sharing a cytomegalovirus(CMV) promoter. The 293 TCymR monoclonal cell strain capable of expressing CymR was screened by Puro resistance and limiting dilution method. The CymR gene position and protein function in the 293T-CymR monoclonal cell strain were verified by PCR, Western blot and fluorescence analysis.Results The pY93-CymR plasmid expressing both the target and resistance genes, was successfully constructed. Following co-transfection with the pX330-CCR5 plasmid into HEK293T cells, four 293T-CymR monoclonal cell strains were obtained via Puro resistance selection and the limiting dilution method. The PCR and Western blot analysis indicated that CymR gene was successfully inserted into the intended CCR5 site and CymR protein was efficiently produced in three of these clones. Subsequent transfection tests demonstrated that the produced CymR could successfully suppress the gene expression under the CMV promoter containing CuO element.Conclusion CymRgene was successfully inserted into the CCR5 locus of HEK293T cell genome, and 293T-CymR monoclonal cell strains capable of expressing CymR and regulating gene expression were obtained.

Extracellular vesicles(EVs)are nanoscale membrane vesicles released by cells that carry bioactive molecules such as proteins and nucleic acids. They play a key role in intercellular communication. In recent years, they have shown significant potential in the diagnosis and treatment of corneal and ocular surface diseases. As diagnostic biomarkers, EVs derived from tears or other sources can reflect disease-specific characteristics, providing a basis for the early disease detection. In terms of therapy, mesenchymal stem cells(MSCs)-derived EVs deliver miRNAs to modulate inflammatory pathways, promote corneal epithelial repair, and inhibit fibrosis. Furthermore, as natural drug vehicles, EVs enable precise delivery of therapeutic agents and promote corneal wound healing. This review systematically summarizes the recent research advances in the application of EVs in the field.

Abnormal blinking can manifest in various ways, including a significant increase or decrease in blink frequency, incomplete blinking, and uncoordinated blinking movements. Currently, a unified diagnostic criterion is still lacking. Abnormal blinking is closely linked to factors such as ocular surface diseases, neurodevelopmental disorders, and visual function abnormalities. In recent years, an increasing number of pediatric patients have visited ophthalmic clinics with abnormal blinking as their primary complaint. With the rising incidence of pediatric meibomian gland dysfunction(MGD)and dry eye disease, their proportion among cases of abnormal blinking has also grown continuously.This article reviews the research progress on meibomian gland changes in children with abnormal blinking. It aims to provide guidance for the early prevention and clinical care of pediatric MGD, reduce the incidence of MGD, and shorten the treatment cycle, thereby providing a solid basis for clinical diagnosis, treatment, and precise prevention.

Herpes zoster (HZ) and its complications remain one of the major public health problems threatening human health. Four HZ vaccines are currently licensed for commercial use worldwide, while novel HZ vaccine candidates based on diverse technological platforms are in different stages of development. Enhancing public understanding of the effectiveness and safety of HZ vaccines is particularly crucial for increasing vaccine coverage and reducing vaccine hesitancy. This article introduces the overview of licensed HZ vaccines with a focus on elucidating the effectiveness and safety of HZ vaccines in immunocompetent and immunocompromised populations, in order to provide a scientific basis for the deployment of HZ vaccination.

Objective To explore the relationship between the oral health status with the severity and outcome of pediatric bronchopneumonia.Methods A total of 150 children patients with bronchopneumonia admitted and treated in this hospital from January 2020 to August 2022 were selected as the observation group(n=150).According to the severity of the illness,the children patients were divided into the mild group(n=97)and severe group(n=53).In addition,150 children undergoing physical examination during the same pe-riod were selected as the control group(n=150).The changes in the decay missing filling surface(DMFS),simplified oral hygiene index(OHI-s)and plaque index(PLI)were analyzed.Spearman was used to analyze the relationship between the oral health status and the severity of bronchopneumonia.After one month of treatment,the changes in oral health status were analyzed under different outcome conditions.The receiver op-erating characteristic(ROC)curve was drawn to assess the predictive effect of the oral health status on the outcome of pediatric bronchopneumonia.Results The DMFS,OHI-s scores and PLI scores in the observation group were higher than those in the control group(P＜0.05).The DMFS,OHI-s score and PLIscores in the severe group were higher than those in the mild group(P＜0.05).Spearman analysis found that DMFS,OHI-s and PLI were positively correlated with the severity(r=0.441,0.253,0.328,P＜0.05).The levels of DMFS,OHI-s and PLI in the poor prognosis group were higher than those in the good prognosis group(P＜0.05).The area under the curve(AUC)of DMFS,OHI-s and PLI combination in predicting the outcome of bronchopneumonia was 0.851(0.703～0.999),with a sensitivity of 83.78％and a specificity of 92.92％.Con-clusion The oral health status is closely related to the severity of pediatric bronchopneumonia,which can be used to assess the outcome of the children patients.

Objective To explore the potential role and underlying mechanism of the functionally uncharacterized gene Gm49394 on regulating β-cell apoptosis under diabetic conditions.Methods The expression and translational activity of Gm49394 in pancreatic β-cell lines and non-β-cell lines were validated using RNA fluorescence in situ hybridization(RNA-FISH),quantitative real-time PCR(qPCR),Western blotting,and immunofluorescence(IF)assay.The β-cell lines(NIT-1/Min6)with Gm49394 overexpression or knockdown were constructed.The proliferation,apoptosis,mitochondrial function,as well as oxidative stress and endoplasmic reticulum stress markers in these β-cell lines under physiological homeostasis or pathological stress conditions,such as high glucose(30 mmol/L),inflammation(10 ng/mL IFN-γ alone or combined with 10 ng/mL IL-6),and hydrogen peroxide(100 μmol/L H2O2)were detected by flow cytometry and Western blotting.Results RNA-FISH and qPCR indicated that Gm49394 was specifically expressed in pancreatic β-cell lines and up-regulated under high glucose or inflammatory stimulation.IF assay and Western blotting showed that Gm49394 had protein-coding activity.Flow cytometry and Western blotting identified that Gm49394 overexpression did not affect β-cell proliferation,but promoted β-cell apoptosis and increased reactive oxygen species(ROS)and mitochondrial superoxide(MitoSOX)levels in β cells under physiological homeostasis or pathological stress conditions(P<0.05).Under physiological conditions,Gm49394 knockdown failed to induce significant alterations on β-cell apoptosis,ROS,or MitoSOX levels.Under pathological stress conditions,Gm49394 knockdown significantly suppressed β-cell proliferation,apoptosis,as well as oxidative and endoplasmic reticulum stress(P<0.05).Conclusion Gm49394 may promote β-cell apoptosis via oxidative stress and endoplasmic reticulum stress.

S-methyl-L-cysteine sulfoxide (SMCO) is a non-protein sulfur-containing amino acid with a variety of functions. There are few reports on the enzymes catalyzing the biosynthesis of SMCO from S-methyl-L-cysteine (SMC). In this study, the flavin-containing monooxygenase gene derived from Schizosaccharomyces pombe (spfmo) was heterologously expressed in Escherichia coli BL21(DE3) and the enzymatic properties of the expressed protein were analyzed. The optimum catalytic conditions of the recombinant SpFMO were 30 ℃ and pH 8.0, under which the enzyme activity reached 72.77 U/g. An appropriate amount of Mg²⁺ improved the enzyme activity. The enzyme kinetic analysis showed that the K_m and k_cat/K_m of SpFMO on the substrate SMC were 23.89 μmol/L and 61.71 L/(min·mmol), respectively. Under the optimal reaction conditions, the yield of SMCO synthesized from SMC catalyzed by SpFMO was 12.31% within 9 h. This study provides reference for the enzymatic synthesis of SMCO.
Schizosaccharomyces/genetics* ; Escherichia coli/metabolism* ; Recombinant Proteins/metabolism* ; Cysteine/biosynthesis* ; Mixed Function Oxygenases/metabolism* ; Schizosaccharomyces pombe Proteins/metabolism* ; Oxygenases/metabolism* ; Kinetics

Objective To investigate the effects of glycerol ingestion on pure tone audiometry(PTA),distor-tion products otoacoustic emission(DPOAE),and electrocochleography(ECochG)in patients with Ménière disease(MD).Methods Glycerol test was conducted in 50 patients with MD.PTA was performed in four series:before glycerol intake,1,2 and 3 hours after intake.DPOAE and ECochG were performed before glycerol intake and 2 hours after intake.All results were analyzed to assess the effect of glycerol on cochlear function of patients with MD.Results ① 55％of MD patients tested positive in PTA glycerol test,and the positive rate increased gradually after 1-3 hours of glycerin ingestion(P＜0.05).For the 33 positive ears,the pure tone threshold decreased the most between 1-2 hours and reached the lowest thresholds at 3 hours.Thresholds at 0.5 kHz,1 kHz,2 kHz dropped the most.② The positive rate of DPOAE glycerol test was 56.67％,with 34 positive ears showing a sig-nificant increase in amplitude between 0.75-2 kHz of f2.③ The positive rate of ECochG glycerin test was 13.64％.The decrease of-SP/AP ratio was not statistically significant before and after ingestion of glycerin(P＞0.05).Conclusion Ingestion of glycerin could alter to varying degrees of the results of PTA,DPOAE and ECo-chG,and influence the cochlear function to some extent.

Objective To evaluate the clinical efficacy and safety of Huoxue Jiedu Prescription in the treatment of polycythemia vera with heat toxicity and blood stasis syndrome.Methods Totally 155 patients of polycythemia vera with heat toxicity and blood stasis syndrome from 5 hospitals including Langfang Traditional Chinese Medicine Hospital from October 2022 to March 2024 were collected.Patients were divided into an observation group(79 cases)and control group(76 cases)using a random number table method.Both groups received conventional Western medicine treatment.The observation group was given Huoxue Jiedu Prescription,one dosage per day,taken orally twice a day;both groups received one treatment course of one month,and three treatment courses were observed.The efficacy of Western medicine and TCM syndromes was observed,and the total symptom assessment scale of myeloproliferative neoplasms(MPN-10)scores,hematological indicators,coagulation function before and after treatment were compared.The safety indicators of the two groups were monitored.Results The control group and observation group lost 2 and 4 cases,respectively.The total effective rate of Western medicine in the observation group was 90.67%(68/75),while the control group was 67.57%(50/74),with statistical significance(P<0.01).The total effective rate of TCM syndromes in the observation group was 94.67%(71/75),while in the control group was 71.62%(53/74),with statistical significance(P<0.01).Compared with before treatment,the total score of MPN-10 in both groups significantly decreased(P<0.05);after treatment,the total score of MPN-10 in the observation group was lower than that in the control group(P<0.05).Compared with before treatment,both groups showed significant reductions in hemoglobin,white blood cell count,hematocrit and platelet count after treatment(P<0.05);after treatment,the above hematological indicators in the observation group were better than those in the control group(P<0.05).Compared with before treatment,the levels of D-dimer and fibrinogen in both groups significantly decreased after treatment,and the activated partial thromboplastin time and prothrombin time were significantly shortened(P<0.05);after treatment,the observation group showed better improvement in the coagulation function indicators compared to the control group(P<0.05).There were no significant adverse reactions in the two groups.Conclusion Huoxue Jiedu Prescription can improve clinical efficacy of polycythemia vera with heat toxicity and blood stasis syndrome,improve hematological indexes,reduce coagulation indexes,and has good safety.

In recent years, the prevalence of myopia has shown a significant upward trend characterized by earlier onset and accelerated progression rates. This epidemic not only imposes an increasing socioeconomic burden but also leads to severe vision impairment through high myopia-related complications that profoundly affect daily life. Current research on the pathogenesis of myopia primarily focuses on four mechanistic theories, including scleral remodeling, choroidal hemodynamic abnormalities, dopamine synthesis and metabolism, and inflammatory responses. The advent of high-throughput sequencing technologies has revolutionized our investigative approaches, enabling deeper exploration into myopia development through multi-omics strategies encompassing genomics, proteomics, and metabolomics. These cutting-edge methodologies have provided novel insights for myopia prevention and control, while simultaneously identifying potential therapeutic targets for precision intervention. This review focuses on summarizing the aforementioned research findings.