BACKGROUND:Synovitis is involved in all stages of osteoarthritis and is a key factor contributing to the development of osteoarthritis.Studies have shown that circular RNA(circRNA)plays an important role in the proliferation,apoptosis and extracellular matrix degradation of synovial cells and chondrocytes.OBJECTIVE:To observe the effects of circRNA SEC24A on the interleukin-1β-induced proliferation,apoptosis,and expression of inflammatory factors in human synovial fibroblasts.METHODS:Human synovial fibroblasts were divided into four groups,including control group,interleukin-1β group,empty vector group,and sh-circSEC24A group.Except for the control group,the other three groups were induced with 10 ng/mL interleukin-1β for 24 hours to establish inflammatory cell models;the empty vector group and sh-circSEC24A group were infected with empty vector virus and lentiviral vector knocking down circSEC24A.CCK-8 assay was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.ELISA was used to detect the levels of matrix metalloproteinase-9,matrix metalloproteinase-13,interleukin-6,and tumor necrosis factor-α in cell supernatant.Western blot assay was used to detect the relative expression levels of Bax,Bcl-2,matrix metalloproteinase-9,matrix metalloproteinase-13,casepase3,cleaved-casepase3,casepase8,and cleaved-casepase8 proteins in cells.RESULTS AND CONCLUSION:(1)qRT-PCR results showed that compared with the normal group,the expression of circSEC24A in human synovial fibroblasts induced by interleukin 1β was significantly up-regulated.(2)The absorbance value of cells in the sh-circSEC24A group detected by CCK-8 assay was significantly higher than that of interleukin 1β group and empty vector group(P＜0.05).The apoptosis rate of sh-circSEC24A group detected by flow cytometry was significantly lower than that of interleukin 1β group and empty vector group(P＜0.05).(3)The levels of tumor necrosis factor α and interleukin 6 in the supernatant of human synovial fibroblasts in the sh-circSEC24A group detected by ELISA were significantly lower than those in the interleukin 1β group and the empty vector group(P＜0.01,P＜0.001).(4)Western blot assay results showed that compared with the interleukin 1β group and the empty vector group,the expression of the pro-apoptotic factor Bax protein in the sh-circSEC24A group significantly decreased,and the expression of the anti-apoptotic factor Bcl-2 protein increased significantly(P＜0.05);apoptosis and related activating factors cleaved-casepase3 and cleaved-casepase8 protein expressions were both reduced(P＜0.05).(5)ELISA and western blot assay results showed that compared with the interleukin 1β group and the empty vector group,the sh-circSEC24A group had lower levels of matrix metalloproteinase 9 and matrix metalloproteinase 13 protein(P＜0.05).These findings indicated that the expression of circSEC24A was abnormally increased in human synovial fibroblasts induced by interleukin 1β.Knocking down circSEC24A expression could promote the proliferation of human synovial fibroblasts and inhibit apoptosis,inflammatory factor release,and extracellular matrix degradation,suggesting that circSEC24A may be an important intervention target for early osteoarthritis.

BACKGROUND:Synovitis is involved in all stages of osteoarthritis and is a key factor contributing to the development of osteoarthritis.Studies have shown that circular RNA(circRNA)plays an important role in the proliferation,apoptosis and extracellular matrix degradation of synovial cells and chondrocytes.OBJECTIVE:To observe the effects of circRNA SEC24A on the interleukin-1β-induced proliferation,apoptosis,and expression of inflammatory factors in human synovial fibroblasts.METHODS:Human synovial fibroblasts were divided into four groups,including control group,interleukin-1β group,empty vector group,and sh-circSEC24A group.Except for the control group,the other three groups were induced with 10 ng/mL interleukin-1β for 24 hours to establish inflammatory cell models;the empty vector group and sh-circSEC24A group were infected with empty vector virus and lentiviral vector knocking down circSEC24A.CCK-8 assay was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.ELISA was used to detect the levels of matrix metalloproteinase-9,matrix metalloproteinase-13,interleukin-6,and tumor necrosis factor-α in cell supernatant.Western blot assay was used to detect the relative expression levels of Bax,Bcl-2,matrix metalloproteinase-9,matrix metalloproteinase-13,casepase3,cleaved-casepase3,casepase8,and cleaved-casepase8 proteins in cells.RESULTS AND CONCLUSION:(1)qRT-PCR results showed that compared with the normal group,the expression of circSEC24A in human synovial fibroblasts induced by interleukin 1β was significantly up-regulated.(2)The absorbance value of cells in the sh-circSEC24A group detected by CCK-8 assay was significantly higher than that of interleukin 1β group and empty vector group(P＜0.05).The apoptosis rate of sh-circSEC24A group detected by flow cytometry was significantly lower than that of interleukin 1β group and empty vector group(P＜0.05).(3)The levels of tumor necrosis factor α and interleukin 6 in the supernatant of human synovial fibroblasts in the sh-circSEC24A group detected by ELISA were significantly lower than those in the interleukin 1β group and the empty vector group(P＜0.01,P＜0.001).(4)Western blot assay results showed that compared with the interleukin 1β group and the empty vector group,the expression of the pro-apoptotic factor Bax protein in the sh-circSEC24A group significantly decreased,and the expression of the anti-apoptotic factor Bcl-2 protein increased significantly(P＜0.05);apoptosis and related activating factors cleaved-casepase3 and cleaved-casepase8 protein expressions were both reduced(P＜0.05).(5)ELISA and western blot assay results showed that compared with the interleukin 1β group and the empty vector group,the sh-circSEC24A group had lower levels of matrix metalloproteinase 9 and matrix metalloproteinase 13 protein(P＜0.05).These findings indicated that the expression of circSEC24A was abnormally increased in human synovial fibroblasts induced by interleukin 1β.Knocking down circSEC24A expression could promote the proliferation of human synovial fibroblasts and inhibit apoptosis,inflammatory factor release,and extracellular matrix degradation,suggesting that circSEC24A may be an important intervention target for early osteoarthritis.

BACKGROUND:Currently,there is no drug that can completely cure osteoarthritis and its pathogenesis is still unclear.Circular RNAs(circRNAs)are differentially expressed in patients with osteoarthritis and are closely associated with various pathological processes in osteoarthritis.circRNAs play an important role in various physiological and pathological processes,such as chondrocyte homeostasis,extracellular matrix formation,and inflammatory response. OBJECTIVE:To mainly review the effects of circRNAs on pathological factors related to osteoarthritis,as well as the types and expression levels of circRNAs in osteoarthritis. METHODS:Related articles published from 1976 to August 2023 were retrieved from CNKI,WanFang,VIP,PubMed,Medline,Web of Science and Elsevier databases.The keywords were"osteoarthritis,circular RNA,non-coding RNA,synovial tissue,chondrocytes"in Chinese and English,respectively.All the relevant articles were screened,summarized,analyzed,and finally 69 papers were included in the review. RESULTS AND CONCLUSION:circRNAs are non-coding RNAs widely found in eukaryotic cells,with covalently closed continuous loop structure,but with no 5'hat structure and 3'poly A tail,which are involved in multi-gene and multi-target regulatory networks and cannot be degraded by nucleic acid exonucleases(RNase R).circRNAs have a high abundance,high conservativeness and stability,and cell and tissue specificity.circRNAs have biological functions such as acting as molecular sponges for miRNAs,regulating linear RNA transcription and RNA shearing,interacting with RNA-bound proteins,and translating proteins.circRNAs regulate chondrocyte apoptosis and proliferation,degradation of cartilage extracellular matrix,and inflammation and other physiopathologic processes.circRNAs are expected to become biomarkers and potential therapeutic targets for clinical diagnosis and prognosis of osteoarthritis,and may become a new strategy for clinical treatment of osteoarthritis in the future.

Objective : To evaluate the vertical distance between the maxillary first molars (MFMs) and the maxillary sinus floor (MSF) and its interrelationship with sex, age, and vertical facial pattern in skeletal ClassⅡ patients to provide a reference for clinical orthodontic treatment. Methods: Sixty teenagers and sixty adults with skeletal Class Ⅱ malocclusion who met the inclusion criteria were selected to evaluate the vertical relationship between the MFMs and the MSF on cone-beam CT (CBCT) images. The vertical distance between the roots of the MFMs and the MSF was measured. Statistical analysis was used to assess differences between patients by sex, age, and vertical facial pattern. Results: The contact percent of the roots of MFMs and MSF was 85% and 56% in skeletal Class Ⅱ teenagers and adults, respectively. The contact percent and penetration percent of the roots with MSF were higher in teenagers than in adults（P<0.05）. The penetration percent of the high-angle (HA) and the normal-angle(NA) groups was 34.1% and 36.6% respectively, which was significantly higher than that in the low-angle (LA) group(20.8%)（P<0.05）. The difference between the distance of the bilateral MFMs and the MSF was not significant in skeletal Class Ⅱ patients (P>0.05); No significant difference was found between different sexes of skeletal Class Ⅱ patients when comparing the distance of the MFMs and the MSF (P>0.05). The MFMs of skeletal Class Ⅱ teenagers were closer to the MSF than those of adults (P<0.05). In the adult group, the distance was not significantly different in different vertical facial patterns (P>0.05). In the teenager group, the MFMs were more closely related to the MSF in the NA and HA groups than in the LA group. Among them, the difference between the mesiobuccal roots and distalbuccal roots was significantly different (P<0.05). There was no significant difference between the groups of the palatal roots (P>0.05). Conclusion The MFMs were closer to the MSF in skeletal Class Ⅱ teenagers than in adults. The distance between the MFMs and MSF was associated with the vertical facial pattern in skeletal Class Ⅱ teenagers, while it was not associated with the vertical facial pattern in adult patients.

Objective In this study ,we constructed a series of recombinant plasmids carriers expressing shRNA targeting hTERT and Bi‐1 gene .These recombinant plasmids carriers were transfected into CNE‐2Z cell lines using Lip and continuously in‐duced the expression of shRNAs .Furthermore ,the shRNAs caused the degradation of mRNAs homologous in sequence with the target genes ,which lead to a sequence‐specific gene silencing .Methods The CNE‐2Z cells was divided into untreated group ,pEG‐FP‐N1 group and pEGFP‐N1/Lip group .Flow cytometry(FCM ) was applied to determine the transfection efficiency .The changes of hTERT and Bi‐1 gene expression were detected by Real‐time RT‐PCR and Western blotting .Results The best transfection effi‐ciency between plasmid and Lip was 2 .5 μg plasmid and 6 .25μL Lip .Conclusion We constructed several shRNA recombinant eu‐karyotic expression plasmids successfully .The recombinant plasmid can inhibit the expression of hTERT and Bi‐1 gene specifically and effectively .

Objective To investigate the way that nasopharyngeal carcinoma (NPC) and NPC stem cells develops resistance to cisplatin through anti-reactive oxygen species mechanism. Methods Using CCK-8 cell counting kit, we measured the half inhibitory concentration of cisplatin against NPC cellsCNE-2and NPC stem cellsCNE-2S, and compared their resistant index. We examined the differences in the reactive oxygen species (ROS) levels, total glutathi?one (GSH) levels, and total superoxide dismutase (SOD) levels between CNE-2 and CNE-2S at different concentrations of cisplatin administration (0.1,0.5 and 1.0μmol·L-1). Using q-PCR, we determined the mRNA expression level of GSS, GCLC, GCLM, SOD1 and SOD2 after 48 hours administration of cisplatin at 1 μmol · L-1. Protein expression level of SOD2 was also tested using Western Blot after 48 hours administration of cisplatin at 1μmol · L-1. Upon silencing the SOD2 in NPC cell through siRNA, Trypan blue was used to analyze cell survival after cisplatin was administrated at 1μmol · L-1. Results The inhibition concentration of cisplatin against CNE-2 was higher than that against CNE-2S (μmol · L-1:9.8 ± 1.1 vs 2.4 ± 0.6,P<0.05). ROS levels in CNE-2 and CNE-2S both rise with cisplatin administration, but ROS levels of CNE-2 before and after cisplatin treatment were both higher than those in CNE-2S (P<0.05). The total gluta?thione levels in CNE-2 and CNE-2S were both increased after 1μmol·L-1 cisplatin treatment but there is no significant dif?ference in levels of glutathione between these two cell lines. After treated with cisplatin, SOD level were increased in both CNE-2S and CNE-2, but it is higher in CNE-2S than that in CNE-2 (P<0.05). The mRNA levels of GSS, GCLC, GCLM, and SOD1 were not different significantly between in CNE-2 and in CNE-2S with or without cisplatin treatment. However, SOD2 in CNE-2S were higher than that in CNE-2 on both mRNA and protein levels (P<0.05). Silenced SOD2 disrupted the resistance of cisplatin in CNE-2S. Conclusion These data suggest that NPC stem cells (CNE-2S) enhance its drug re?sistance to cisplatin through highly expression of SOD2 which posed anti-ROS capacity.

Objective To investigate the role of ATP￣binding cassette ( ABC ) family on the resistance of nasopharyngeal carcinoma (NPC) stem cells (CSCs) to cisplatin. Methods We compared the differences between the drug extravasation capability of CNE￣2 and CNE￣2S by using Rhodamine￣123 efflux assay. We determined the mRNA and protein expression levels of ABC transport family members, including ABCA3,ABCB1,ABCB5,ABCC1,ABCC2 and ABCG2,after 48 h being treated with 1 μmol.L-1 cisplatin by RT￣PC and Western blotting.Rhoamine￣123 efflux and apoptosis by cisplatin in two kinds of cells was examined by ABCA3 gene silencing with specific small￣interfering RNA. Results The IC50 of cisplatin on CNE￣2S was 4.1 fold to that on CNE￣2(P<0.05).For the relative drug effluent activity and Na+K+ ATPase activity,CNE￣2S was 4.8 fold to CNE￣2(P<0.05),suggested that CNE￣2S expressed more ABCA3,ABCB1,ABCC1 and ABCG2 in comparison to CNE￣2(P<0.05).After 48 h treatment with 1 μmol.L-1 cisplatin,ABCA3 specifically highly expressed in CNE￣2S (P<0.05), and knocking down of ABCA3 resulted in reduction of rhodamine￣123 efflux and increase of apoptosis. Conclusion The cisplatin resistance of NPC CSCs is associated with enhanced expression of ABCA3,ABCC1 and ABCG2, suppression of ABCA3 could reverse the resistance of NPC CSCs to cisplatin.

Objective To explore the influence of intensive glycemic control by insulin pump on prognosis of critically ill patients with lung infection and respiratory failure.Methods In the emergency intensive care unit (EICU),200 critically ill patients with hyperglycemia (A-PACHE II score >15 ,random blood glucose > 11.1 mmol /L)were collected and randomly di-vided into the intensive insulin therapy (IIT)group and the convention insulin therapy(CIT) group(use insulin pump to control blood glucose).IIT group and CIT group included 31 cases and 33 cases of pulmonary infection and respiratory failure.Ventilator and antibiotic use days,short-term mortality (within 28 days),rate of hypoglycemia,nosocomial infection,hospital stay and hospital costs were observed and compared between two groups.Results There was no significant differences between two groups in aspects of age,sex ratio,oxygen saturation,pressure of oxygen, pressure of Carbon dioxide,pH,blood pressure,respiratory failure type,blood glucose,elec-trolytes,inflammation,heart function,liver and kidney function,fasting C peptide,HbAlc and APACHE Ⅱ score(P >0.05).APACHE II score at the time of 3 days and 7 days after admission, nosocomial infection,short-term mortality hospital day and hospital costs (P <0.05)in the IIT group were significantly lower and shorter than the CIT group.The hypoglycemia incidence rate of the IIT group was significantly higher than that of the CIT group (P <0.01).Result of serious hypoglycemia showed no significant difference between the two groups .Conclusion Strict intensive glucose control on pulmonary infection and respiratory failure may bring more benefits for acute and critically ill patients,and it can reduce the short-term mortality rate.

Objective To explore the influence of intensive glycemic control by insulin pump on prognosis of critically ill patients with lung infection and respiratory failure.Methods In the emergency intensive care unit (EICU),200 critically ill patients with hyperglycemia (A-PACHE II score >15 ,random blood glucose > 11.1 mmol /L)were collected and randomly di-vided into the intensive insulin therapy (IIT)group and the convention insulin therapy(CIT) group(use insulin pump to control blood glucose).IIT group and CIT group included 31 cases and 33 cases of pulmonary infection and respiratory failure.Ventilator and antibiotic use days,short-term mortality (within 28 days),rate of hypoglycemia,nosocomial infection,hospital stay and hospital costs were observed and compared between two groups.Results There was no significant differences between two groups in aspects of age,sex ratio,oxygen saturation,pressure of oxygen, pressure of Carbon dioxide,pH,blood pressure,respiratory failure type,blood glucose,elec-trolytes,inflammation,heart function,liver and kidney function,fasting C peptide,HbAlc and APACHE Ⅱ score(P >0.05).APACHE II score at the time of 3 days and 7 days after admission, nosocomial infection,short-term mortality hospital day and hospital costs (P <0.05)in the IIT group were significantly lower and shorter than the CIT group.The hypoglycemia incidence rate of the IIT group was significantly higher than that of the CIT group (P <0.01).Result of serious hypoglycemia showed no significant difference between the two groups .Conclusion Strict intensive glucose control on pulmonary infection and respiratory failure may bring more benefits for acute and critically ill patients,and it can reduce the short-term mortality rate.

Objective: The present study aimed to isolate and characterize a cancer stem cell-like sphere-forming cell subpopula-tion. Methods: By using a spheroid culture stem cell-conditioned medium, we isolated a subgroup of cancer stem-like cells from naso-pharyngeal cancer cell lines. Chemotherapy resistance was analyzed by using methyl thiazolyl tetrazolium, and clone-forming capabili-ty was determined by using softer agar clone formation assay. Finally, we verified the expression of the stemness-specific gene andβ-catenin by using immunocytochemistry staining and RT-PCR. Results: The lower-differentiated nasopharyngeal cancer lines con-tained more sphere-forming cells, whereas sphere-forming cells were not observed in the high differentiated nasopharyngeal cancer line CNE-1. Compared with CNE-2, CNE-2S (sphere-forming cells derived from CNE-2) exhibited higher chemotherapy resistance and clone-forming ability. Interestingly, the stemness genes BMI-1, ABCG2, and ALDH1 exhibited higher expression in CNE-2S than in CNE-2. β-catenin, a vital transcription factor of the WNT pathway related to stem cells, exhibited higher expression in CNE-2s cellular nucleus and plasma than in CNE-2. Conclusion: We isolated a subgroup of stem-like nasopharyngeal cancer sphere-forming cells. This discovery paves the way for the development of therapeutic strategies aimed at eradicating tumorigenic subpopulations in nasopharyn-geal cancer.