Objective: To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1 cells, and to clarify its mechanism Methods: Three kinds of human esophageal cancer cells, TE-1, ECA109 and KYSE150, were selected. The relative expression levels of MRPL35 mRNA in three kinds of cells by real-time quantitative PCR. The esophageal cancer TE-1 cells were divided into sh MRP 1.35 group and shCtrl group, and the cells were infcctcd with si-RNA lentivirus and si-RNA lentivirus; the esophageal cancer cell line stably silenting the MRPL35 gene was established. Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35 gene silencing. The cell growth curves in various groups were detected by CCK-8 method, and the apoptotic rates were detected by flow cytometry after Annexin \ -PE/7AAD double staining. Results: Three kinds of esophageal cancer cells expressed MRPL35 gene, and the expression levels were not statistically significant between them (P>0.05). The results of real-time quantitative PCR and Western blotting methods showed that the mRNA and protein levels of MRPL35 in the TE-1 cells in shMRPI.35 group were significantly lower than those in shCtrl group ( P<∗0. 05). Compared with shCtrl group, the cell growth speed in shMRPI.35 group was decreased ( P

Objective:To observe the efficacy and safety of recombinant human thrombopoietin (rhTPO) in the treatment of radiation induced thrombocytopenia (RIT) .Methods:From January 2019 to March 2021, 204 cases (including 101 cases of radiotherapy alone and 103 cases of concurrent chemoradiotherapy) were collected retrospectively after radiotherapy and with decreased in blood platelet count <75×10 9/L in Jilin Cancer Hospital. These patients received rhTPO 15 000 U, once a day, subcutaneous, for at least 4 consecutive days, or met the withdrawal criteria blood platelet count ≥100×10 9/L, or the absolute value of blood platelet increase ≥50×10 9/L. The characteristics of blood platelet decline, treatment efficacy, and safety were analyzed. Results:The numbers of radiotherapy treatments with platelets lower than 75×10 9/L in the radiotherapy alone group and the concurrent chemoradiotherapy group were 19 (13, 22) and 13 (10, 17) times, respectively, indicating that patients in the concurrent chemoradiotherapy group experienced platelet decline earlier ( Z=-5.27, P<0.001), the lowest values of platelet decline in the two groups were 68 (45, 74) ×10 9/L and 62 (44, 74) ×10 9/L, respectively, with no statistically significant difference ( Z=-1.15, P=0.252). After received rhTPO treatment, the numbers of days that the two groups of patients had platelets <50×10 9/L were 7 (3, 13) d and 7 (5, 11) d, respectively, with no statistically significant difference ( Z=-1.13, P=0.281). After the patients received radiotherapy, rhTPO was started when the platelet count dropped to <75×10 9/L. The number of days required to recover to 75×10 9/L was 4 (2, 10) d in the radiotherapy alone group and 4 (2, 8) d in the concurrent chemoradiotherapy group, with no statistically significant difference ( Z=-1.07, P=0.285) ; the number of days required for platelets to recover to 100×10 9/L or for the absolute value to increase by 50×10 9/L was 8 (6, 14) d in the radiotherapy alone group and 11 (8, 16) d in the concurrent chemoradiotherapy group. The recovery time of the concurrent chemoradiotherapy group was longer than that of the radiotherapy alone group ( Z=-3.64, P<0.001). Regardless of the baseline level, there was no statistically significant difference in the number of days for platelets to recover to 75×10 9/L after rhTPO treatment between the radiotherapy alone group and the concurrent chemoradiotherapy group ( Z=-1.42, P=0.155; Z=-0.97, P=0.332). The number of days required for the two groups of patients to recover to 100×10 9/L or for the absolute value to increase by 50×10 9/L were 8 (6, 14) d and 11 (8, 16) d, respectively, with a statistically significant difference ( Z=-3.64, P<0.001). The numbers of days required for the two groups of patients with baseline platelets ≥50×10 9/L to recover to 100×10 9/L or for the absolute value to increase by 50×10 9/L were 8 (4, 12) d and 10 (8, 16) d, respectively, with a statistically significant difference ( Z=-3.12, P=0.002). However, there was no statistically significant difference in the number of days required for the two groups of patients with baseline platelets <50×10 9/L to recover to 100×10 9/L or for the absolute value to increase by 50×10 9/L ( Z=-1.88, P=0.061). The total platelet elevation rate of rhTPO within 20 days of radiotherapy treatment for both groups of patients was 93.63% (191/204), of which 95.05% (96/101) was for radiotherapy alone and 92.23% (95/103) for concurrent chemoradiotherapy, with no statistically significant difference ( χ2=0.68, P=0.410). In addition, there was no statistically significant difference in gender ( χ2=3.47, P=0.063), age ( χ2=2.79, P=0.095), TNM staging ( χ2=5.07, P=0.167), and baseline platelet count ( χ2=0.62, P=0.822) between the two groups.During the radiotherapy cycle, 27 patients (13.23%) received blood platelet infusion, and 158 patients (77.45%) completed the radiotherapy plan without interruption. No rhTPO-related adverse reactions were found. Conclusion:rhTPO in the treatment for RIT can effectively promote the recovery of blood platelet without any adverse reactions, and has good safety.

Objective:To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1cells, and to clarify its mechanism.Methods:Three kinds of human esophageal cancer cells, TE-1, ECA109and KYSE150, were selected.The relative expression levels of MRPL35mRNA in three kinds of cells by real-time quantitative PCR.The esophageal cancer TE-1cells were divided into shMRPL35group and shCtrl group, and the cells were infected with si-RNA lentivirus and si-RNA lentivirus;the esophageal cancer cell line stably silenting the MRPL35gene was established.Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35gene silencing.The cell growth curves in various groups were detected by CCK-8method, and the apoptotic rates were detected by flow cytometry after AnnexinⅤ-PE/7AAD double staining.Results:Three kinds of esophageal cancer cells expressed MRPL35gene, and the expression levels were not statistically significant between them (P＞0.05) .The results of real-time quantitative PCR and Western blotting methods showed that the mRNA and protein levels of MRPL35in the TE-1cells in shMRPL35group were significantly lower than those in shCtrl group (P＜0.05) .Compared with shCtrl group, the cell growth speed in shMRPL35group was decreased (P＜0.05) , and the apoptotic rate was significantly increased (P＜0.01) .Conclusion:Silencing MRPL35gene can inhibit the proliferation of esophageal cancer TE-1cells and plays a role through the apoptotic pathway.

Nucleic acid-based drugs, such as RNA and DNA drugs, exert their effects at the genetic level. Currently, widely utilized nucleic acid-based drugs include nucleic acid aptamers, antisense oligonucleotides, mRNA, miRNA, siRNA and saRNA. However, these drugs frequently encounter challenges during clinical application, such as poor stability, weak targeting specificity, and difficulties in traversing physiological barriers. By employing chemical modifications of nucleic acid structures, it is possible to enhance the stability and targeting specificity of certain nucleic acid drugs within the body, thereby improving delivery efficiency and reducing immunogenicity. Moreover, utilizing nucleic acid drug carriers can facilitate the transportation of drugs to lesion sites, thereby aiding efficient intracellular escape and promoting drug efficacy within the body. Currently, commonly employed delivery carriers include virus vectors, lipid nanoparticles, polymer nanoparticles, inorganic nanoparticles, protein carriers and extracellular vesicles. Nevertheless, individual modifications or delivery carriers alone are insufficient to overcome numerous obstacles. The integration of nucleic acid chemical modifications with drug delivery systems holds promise for achieving enhanced therapeutic effects. However, this approach also presents increased technical complexity and clinical translation costs. Therefore, the development of nucleic acid drug carriers and nucleic acid chemical modifications that are both practical and simple, while maintaining high efficacy, low toxicity, and precise nucleic acid delivery, has become a prominent research focus in the field of nucleic acid drug development. This review comprehensively summarizes the advancements in nucleic acid-based drug modifica-tions and delivery systems. Additionally, strategies to enhance nucleic acid drug delivery efficiency are discussed, with the aim of providing valuable insights for the translational application of nucleic acid drugs.
Nucleic Acids ; RNA, Small Interfering/genetics* ; Drug Carriers ; Drug Delivery Systems ; Drug Development

Mitochondrial DNA (mtDNA) mutations result in a variety of genetic diseases. As an emerging therapeutic method, mtDNA editing technology recognizes targets more based on the protein and less on the nucleic acid. Although the protein recognition type mtDNA editing technology represented by zinc finger nuclease technology, transcription activator like effector nuclease technology and base editing technology has made some progress, the disadvantages of complex recognition sequence design hinder further popularization. Gene editing based on nucleic acid recognition by the CRISPR system shows superiority due to the simple structure, easy design and modification. However, the lack of effective means to deliver nucleic acids into mitochondria limits application in the field of mtDNA editing. With the advances in the study of endogenous and exogenous import pathways and the deepening understanding of DNA repair mechanisms, growing evidence shows the feasibility of nucleic acid delivery and the broad application prospects of nucleic acid recognition type mtDNA editing technology. Based on the classification of recognition elements, this article summarizes the current principles and development of mitochondrial gene editing technology, and discusses its application prospects.
Genes, Mitochondrial ; Gene Editing ; Mitochondria/genetics* ; DNA, Mitochondrial/genetics* ; Nucleic Acids ; Technology

Long non-coding RNAs (lncRNAs) which are usually thought to have no protein coding ability, are widely involved in cell proliferation, signal transduction and other biological activities. However, recent studies have suggested that short open reading frames (sORFs) of some lncRNAs can encode small functional peptides (micropeptides). These micropeptides appear to play important roles in calcium homeostasis, embryonic development and tumorigenesis, suggesting their potential as therapeutic targets and diagnostic biomarkers. Currently, bioinformatic tools as well as experimental methods such as ribosome mapping and in vitro translation are applied to predict the coding potential of lncRNAs. Furthermore, mass spectrometry, specific antibodies and epitope tags are used for validating the expression of micropeptides. Here, we review the physiological and pathological functions of recently identified micropeptides as well as research strategies for predicting the coding potential of lncRNAs to facilitate the further research of lncRNA encoded micropeptides.
Female ; Pregnancy ; Humans ; RNA, Long Noncoding/genetics* ; Research Design ; Antibodies ; Carcinogenesis ; Micropeptides

Immunological evasion is one of the defining characteristics of cancers, as the immune modification of an immune checkpoint (IC) confers immune evasion capabilities to tumor cells. Multiple ICs, such as programmed cell death protein-1 (PD-1) and cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4), can bind to their respective receptors and reduce tumor immunity in a variety of ways, including blocking immune cell activation signals. IC blockade (ICB) therapies targeting these checkpoint molecules have demonstrated significant clinical benefits. This is because antibody-based IC inhibitors and a variety of specific small molecule inhibitors can inhibit key oncogenic signaling pathways and induce durable tumor remission in patients with a variety of cancers. Deciphering the roles and regulatory mechanisms of these IC molecules will provide crucial theoretical guidance for clinical treatment. In this review, we summarize the current knowledge on the functional and regulatory mechanisms of these IC molecules at multiple levels, including epigenetic regulation, transcriptional regulation, and post-translational modifications. In addition, we provide a summary of the medications targeting various nodes in the regulatory pathway, and highlight the potential of newly identified IC molecules, focusing on their potential implications for cancer diagnostics and immunotherapy.
Apoptosis Regulatory Proteins ; CTLA-4 Antigen/therapeutic use* ; Epigenesis, Genetic ; Humans ; Immunotherapy ; Neoplasms/therapy* ; Programmed Cell Death 1 Receptor/therapeutic use*

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Humans ; Apoptosis Inducing Factor/metabolism* ; NAD/metabolism* ; Dimerization ; Apoptosis