ObjectiveTo explore the interaction and competitive binding of Homo sapiens long intergenic non-protein-coding RNA 1134 （LINC01134） to CCCTC-binding factor CTCF， affecting the transcription of cyclin-dependent kinase inhibitor （p21） and influencing the proliferation of A549 cells， in order to investigate the possible mechanism of Astragali Radix and Hedyotis diffusa （A-H） in inhibiting A549 proliferation by regulating this axis. MethodsRNA-binding protein immunoprecipitation （RIP） assays were conducted to examine the interaction between LINC01134 and CTCF， and chromatin immunoprecipitation （ChIP） assays were used to study the effect of LINC01134 overexpression on the interaction between CTCF and p21. Stable A549 cell lines （oe-NC and oe-LINC01134） were established using lentiviral transfection， and each group was treated with 10% A-H drug-containing serum. Real-time PCR and Western blot analyses were performed to detect the effects of A-H on the expression of LINC01134， CTCF， and p21 in A549 cells. Cell counting kit-8 （CCK-8） and colony formation assays were used to assess the effects of A-H on A549 cell proliferation via LINC01134. Flow cytometry was employed to evaluate the effects of A-H on the A549 cell cycle through LINC01134， and Western blot was used to detect changes in cell cycle proteins. ResultsCompared with the IgG group， the oe-CTCF group showed a significantly increased abundance of LINC01134 aggregates （P0.01）. Compared with the oe-Vector group， p21 abundance in CTCF complexes was significantly reduced in the oe-LINC01134 group （P0.01）. Compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed the expression of LINC01134 and p21 （P0.05）， but had no significant regulatory effect on CTCF. Compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed cell viability at 72 h （P0.05）， inhibited malignant proliferation （P0.05）， and reversed the proportions of cells in the G₀/G₁ and S phases （P0.01）. Furthermore， compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed the expression of Cyclin D₁， CDK4， Cyclin E， CDK2， phosphorylated retinoblastoma protein （p-Rb）， and E2F transcription factor 3 （E2F3） （P0.01）. ConclusionA-H regulates the LINC01134-CTCF-p21 axis to block the G₁/S phase transition of A549 cell cycle， accelerate cellular senescence， and inhibit malignant proliferation.

ObjectiveTo explore the interaction and competitive binding of Homo sapiens long intergenic non-protein-coding RNA 1134 （LINC01134） to CCCTC-binding factor CTCF， affecting the transcription of cyclin-dependent kinase inhibitor （p21） and influencing the proliferation of A549 cells， in order to investigate the possible mechanism of Astragali Radix and Hedyotis diffusa （A-H） in inhibiting A549 proliferation by regulating this axis. MethodsRNA-binding protein immunoprecipitation （RIP） assays were conducted to examine the interaction between LINC01134 and CTCF， and chromatin immunoprecipitation （ChIP） assays were used to study the effect of LINC01134 overexpression on the interaction between CTCF and p21. Stable A549 cell lines （oe-NC and oe-LINC01134） were established using lentiviral transfection， and each group was treated with 10% A-H drug-containing serum. Real-time PCR and Western blot analyses were performed to detect the effects of A-H on the expression of LINC01134， CTCF， and p21 in A549 cells. Cell counting kit-8 （CCK-8） and colony formation assays were used to assess the effects of A-H on A549 cell proliferation via LINC01134. Flow cytometry was employed to evaluate the effects of A-H on the A549 cell cycle through LINC01134， and Western blot was used to detect changes in cell cycle proteins. ResultsCompared with the IgG group， the oe-CTCF group showed a significantly increased abundance of LINC01134 aggregates （P0.01）. Compared with the oe-Vector group， p21 abundance in CTCF complexes was significantly reduced in the oe-LINC01134 group （P0.01）. Compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed the expression of LINC01134 and p21 （P0.05）， but had no significant regulatory effect on CTCF. Compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed cell viability at 72 h （P0.05）， inhibited malignant proliferation （P0.05）， and reversed the proportions of cells in the G₀/G₁ and S phases （P0.01）. Furthermore， compared with the 10% blank + oe-LINC01134 group， the 10% A-H + oe-LINC01134 group reversed the expression of Cyclin D₁， CDK4， Cyclin E， CDK2， phosphorylated retinoblastoma protein （p-Rb）， and E2F transcription factor 3 （E2F3） （P0.01）. ConclusionA-H regulates the LINC01134-CTCF-p21 axis to block the G₁/S phase transition of A549 cell cycle， accelerate cellular senescence， and inhibit malignant proliferation.

ObjectiveTo investigate the effects of excessive activation of the Specific protein 1 （Sp1）-MicroRNA-582-3p （miR-582-3p）-cyclin-dependent kinase inhibitor （p27） axis on the cell cycle and proliferation of A549 lung adenocarcinoma cells at the cellular level， thereby investigating the possible mechanisms by which Astragali Radix and Hedyotis diffusa（A-H） regulate the axis to inhibit A549 cells proliferation. MethodsLentiviral plasmid transfection technology was used to obtain four stable A549 transgenic cell lines： shRNA-Sp1+LV-miR-582-3p mimic， shRNA-Sp1+LV-mNC， shRNA-NC+LV-miR-582-3p mimic， and shRNA-NC+LV-mNC. Real-time PCR was used to detect the effect of Sp1 on miR-582-3p expression in A549 lung adenocarcinoma cells. CCK-8 and colony formation assay were used to detect the effect of Sp1 on cell proliferation through miR-582-3p， while flow cytometry was used to detect the effect of Sp1 on cell cycle via miR-582-3p. Western blot was used to detect the effect of Sp1 on cyclin in A549 cells through miR-582-3p. CCK-8 was employed to detect the effects of different concentrations （5%， 10%， 20%， 40%， 80%） of A-H on A549 cell viability， and the optimal concentration was selected for subsequent mechanism exploration. Stable transgenic strains oe-Sp1 and control oe-Vector were constructed using the above-mentioned transfection techniques. Real-time PCR was then used to investigate the effect of A-H on miR-582-3p expression in A549 cells， while Western blot was employed to detect the effect of A-H on Sp1 and p27 expression in A549 lung adenocarcinoma cells. ResultsCompared with the shRNA-NC+LV-mNC group， the expression of miR-582-3p was significantly inhibited in the shRNA-Sp1+LV-mNC group （P<0.01）. Compared with the shRNA-Sp1+LV-mNC group， the shRNA-Sp1+LV-miR-582-3p mimic group significantly reversed miR-582-3p expression （P<0.01）， indicating that Sp1 has a significant regulatory effect on miR-582-3p. Compared with the shRNA-Sp1+LV-mNC group， the shRNA-Sp1+LV-miR-582-3p mimic group significantly reversed cell proliferation （P<0.01）， suggesting that miR-582-3p can partially reverse the proliferative effect of Sp1 on A549 lung adenocarcinoma. Compared with the shRNA-Sp1+LV-mNC group， the shRNA-Sp1+LV-miR-582-3p mimic group showed a significant decrease in G₀/G₁ proportions， while the G₂/M and S phase proportions increased significantly （P<0.01）， indicating that miR-582-3p can partially reverse the effect of Sp1 on the A549 cell cycle. Compared with the shRNA-Sp1+LV-mNC group， the shRNA-Sp1+LV-miR-582-3p mimic group significantly reversed the expression of various proteins with the decrease of p27 and the increase of Cyclin D₁， CDK4， Cyclin E， CDK2， p-Rb， and E2F3 （P<0.01）， indicating that miR-582-3p can partially reverse the effect of Sp1 on the cell cycle of A549 lung adenocarcinoma. Compared with the blank group， the serum containing A-H significantly inhibited A549 cell viability in a concentration-dependent manner. Therefore， 10% A-H was selected for intervention of subsequent mechanism experiments. Compared with A-H+oe-Vector， A-H+oe-Sp1 significantly reversed the downregulation of Sp1， miR-582-3p， and the upregulation of p27 （P<0.01）. This suggests that Sp1 can partially reverse the effects of A-H on miR-582-3p and p27 levels in A549 lung adenocarcinoma cells. ConclusionOveractivation of the Sp1-miR-582-3p-p27 axis significantly promotes the proliferation of A549 lung adenocarcinoma cells， and the potential mechanism of the inhibitory effect of A-H on the proliferation of A549 cells may be related to its inhibition on the overactivation of the Sp1-miR-582-3p-p27 axis.

BACKGROUND:Increasing studies have shown that autophagy plays an important role in the treatment of osteoarthritis,and moderate autophagy can preserve the normal physiological function of osteoarticular chondrocytes.Traditional Chinese medicine(TCM)monomers can target and modulate autophagy to treat osteoarthritis,and their characteristics such as single components,clear efficacy,low price,and easy availability have obvious benefits in the treatment of osteoarthritis.OBJECTIVE:To review the effects of TCM monomers on the targeted regulation of autophagy in the treatment of osteoarthritis and the research progress,with a view to laying a foundation for the treatment of osteoarthritis and even other bone metabolic diseases.METHODS:Relevant literature published from January 2012 to October 2022 was retrieved in PubMed,Web of Science,CNKI,and WanFang using the keywords of"traditional Chinese medicine,Chinese herbal monomer,autophagy,osteoarthritis"in English and Chinese.Inclusion and exclusion criteria were developed,and 63 relevant articles were finally included by screening through reading the title,abstract,and full-text content.RESULTS AND CONCLUSION:TCM monomers can treat osteoarthritis by targeting autophagy to inhibit chondrocyte apoptosis,protect cartilage extracellular matrix,reduce inflammation and antagonize oxidative stress injury.Different TCM monomers can regulate autophagy in the same way,and the same TCM monomers can affect autophagy in different ways.The combination of multiple monomers and the multi-target and multi-pathway regulation of autophagy by TCM monomers remain to be explored.The regulation of autophagy by TCM monomers can provide new ideas and strategies for the prevention and treatment of osteoarthritis.Moderate regulation of autophagy by TCM monomers to keep the autophagic flux unimpeded may be the key to treating osteoarthritis.

BACKGROUND:More and more studies have shown that oxidative stress should play an important role in the treatment of osteoporosis.Oxidative stress should cause the accumulation of oxidation activity,which will damage bone-related cells.Finally,it causes the imbalance of bone resorption and bone formation,resulting in a decrease in bone volume and the destruction of the slight structure.Research in recent years has found that some natural products can regulate oxidative stress to treat osteoporosis.The characteristics of extensive sources and small side effects have obvious advantages in the treatment of osteoporosis,and the efficacy is objective. OBJECTIVE:To discuss the mechanism of natural product regulation of oxidation stress in treatment of osteoporosis,conduct a review based on the latest related research progress,provide reference and ideas for more natural products to treat osteoporosis in the future,and provide data support for the clinical application of natural compounds in the treatment of osteoporosis. METHODS:"oxidative stress,free radical,antioxidant,phytotherapy,plant extracts,medicinal plants,herbal medicine,osteoporosis,bone density,bone loss"were used as the keywords in PubMed,Web of Science,Embase,Cochrane,VIP,CBM,WanFang,and CNKI databases to search relevant articles published from January 2010 to February 2023.Inclusion and exclusion criteria were developed,and 64 relevant articles were selected by reading titles,abstracts,and full texts. RESULTS AND CONCLUSION:(1)Some natural products have antioxidant effects and can regulate osteogenic differentiation,osteoblast bone matrix mineralization,osteoclast-mediated bone resorption,proliferation,differentiation,activity,and apoptosis of bone-related cells by improving oxidative stress,thus affecting bone metabolism.(2)These natural products with antioxidant effects play a role in treating osteoporosis by improving bone remodeling balance.(3)The research on the combination of a variety of natural products to improve osteoporosis remains to be explored.(4)The use of natural products to regulate oxidative stress may become a powerful weapon for the clinical treatment of osteoporosis in the future.

Osteoporosis， a common metabolic bone disease， is caused by the imbalance of bone homeostasis. Bone homeostasis depends on the dynamic balance between bone resorption by osteoclasts and bone matrix formation by mesenchymal lineage osteoblasts and involves a series of complex and highly regulated steps. When bone resorption is faster than bone formation， bone homeostasis will be destroyed， which will lead to osteoporosis. Autophagy is a protein and organelle degradation pathway important for the maintenance of cytoplasmic homeostasis. The basal level of autophagy is present in all the three types of bone cells. Autophagy is a process whereby damaged organelles are recycled and bone remodeling continues， and thus it plays an important role in bone maintenance. Therefore， it is possible to regulate bone metabolism by targeting specific autophagy-related molecules. At the same time， as the research on the treatment of osteoporosis by Chinese medicine advances， it is found that the monomers isolated from traditional Chinese medicinals have the potential of regulating autophagy in the treatment of osteoporosis. Thus， this paper discusses the relationship between autophagy and osteoporosis and summarizes six different mechanisms of 22 Chinese medicine monomers in the treatment of osteoporosis through targeting autophagy： increasing osteogenic differentiation of mesenchymal stem cells， promoting osteoblast mineralization， inhibiting osteoclasts differentiation， reducing apoptosis of bone cells， antagonizing oxidative stress injury， and coordinating inflammation and autophagy levels. The review is expected to provide a reference and ideas for future research on the treatment of osteoporosis by Chinese medicine monomers.

The dental operative microscope has been widely employed in the field of dentistry, particularly in endodontics and operative dentistry, resulting in significant advancements in the effectiveness of root canal therapy, endodontic surgery, and dental restoration. However, the improper use of this microscope continues to be common in clinical settings, primarily due to operators' insufficient understanding and proficiency in both the features and established operating procedures of this equipment. In October 2019, Professor Jingping Liang, Vice Chairman of the Society of Cariology and Endodontology, Chinese Stomatological Association, organized a consensus meeting with Chinese experts in endodontics and operative dentistry. The objective of this meeting was to establish a standard operation procedure for the dental operative microscope. Subsequently, a consensus was reached and officially issued. Over the span of about four years, the content of this consensus has been further developed and improved through practical experience.
Humans ; Dentistry, Operative ; Consensus ; Endodontics ; Root Canal Therapy ; Dental Care

Objective:To investigate the effect of the adaptive computerized cognitive training(ACCT) on patients with mild cognitive impairment(MCI).Methods:A total of 114 patients with mild cognitive impairment (53 cases in the treatment group and 61 cases in the observation group) were selected.In the treatment group, routine treatment combined with ACCT were given for 24 weeks, then routine treatment only for 24 weeks, 48 weeks altogether. In the observation group, routine treatment was given for 48 weeks.At week 0, 24, 48, both groups were assessed by scales including: mini-mental state examination (MMSE), Montreal cognitive assessment(MoCA), numerical memory span test, activities of daily living, Hamilton depression scale(HAMD)and Hamilton anxiety scale(HAMA). SAS 9.4 was used for statistical analysis.The data were analyzed by χ 2 test, rank sum test and multi-level model analysis. Results:Interactions between group and time on MMSE( treatment group: 0 week 22.0 (21.0, 23.0), 24 weeks 24.0 (24.0, 25.0), 48 weeks 25.0 (24.0, 26.0); observation group: 0 week 23.0 (21.0, 24.0), 24 weeks 23.0 (21.0, 24.0), 48 weeks 23.0 (21.0, 24.0)), MoCA( treatment group: 0 week 18.0 (17.0, 20.0), 24 weeks 22.0 (20.0, 23.0), 48 weeks 22.0 (20.0, 24.0); observation group: 0 week 19.0 (17.0, 20.0), 24 weeks 19.0 (18.0, 20.0), 48 weeks 19.0 (18.0, 20.0)), IADL( treatment group: 0 week 11.0 (10.0, 13.0), 24 weeks 12.0 (10.0, 12.0), 48 weeks 12.0 (10.0, 12.0); observation group: 0 week 12.0 (11.0, 13.0), 24 weeks 11.0 (10.0, 12.0), 48 weeks 11.0 (10.0, 12.0)), DST-forwards and HAMD scores were significant(all P<0.05), and DST-backwards had significant group main effect ( P<0.05). Further simple effect analysis showed that the influence of group and time on MMSE, MoCA and DST-forwards were statistically significant (all P<0.05), and the influence of time on IADL and HAMD were statistically significant (both P<0.05). Further comparison of the difference between the two groups at each time point: D-value of MMSE, MoCA, DST-forwards, and DST-backwards score in the treatment group were higher than those in the observation group, while D-value of HAMD score was lower than that in the observation group, and the differences were statistically significant (all P<0.05). Conclusion:ACCT may have long term effect on improving the cognitive function and depression of MCI patients.

Objective:To evaluate anatomical locking plate combined with bone cement intramedullary support in the treatment of proximal humeral fractures in the elderly.Methods:From May 2016 to July 2018, 19 geriatric proximal humeral fractures were treated with anatomical locking plate combined with bone cement intramedullary support at Department of Orthopaedics, General Hospital of Shenyang Northern Theater. There were 8 females and 5 males, with an average age of 71 years (from 60 to 84 years). All the fractures were closed. According to the Neer classification, there were 4 two-part, 5 three-part and 4 four-part fractures. At 6, 12, 18 and 36 weeks and one year postoperatively, anteroposterior and lateral X-ray films of the scapula were taken. At the last follow-up, the Constant-Murley shoulder score and the Disabilities of the Arm, Shoulder and Hand (DASH) score for the upper limb dysfunction were recorded.Results:The 13 patients were followed up for 3 to 12 months (6 months on average). Bony union was achieved in all the 13 patients. Local necrosis and deformation of the humeral head and partial screw perforation were observed one year after operation in 2 cases, but other patients reported no such complications or loosening of internal fixation. At the last follow-up, their mean Constant-Murley score was 82.6 (from 71 to 95) and their mean ADSH score 19.2 (from 9.2 to 48.1).Conclusion:Anatomical locking plate combined with bone cement intramedullary support may lead to fine early efficacy for the treatment of proximal humeral fractures in the elderly, since it can facilitate fracture reduction, promote rigid fixation and prevent loss of reduction.

Objective: To investigate the effect of X-ray fluoroscopy and CT guided technique in sacroiliac screw fixation for type Tile B or C sacrum pelvic fractures or dislocations. Methods: A retrospective case control study was conducted to analyze the clinical data of 103 patients with type Tile B or C posterior pelvic ring fracture or dislocation admitted to the General Hospital from Northern Theater of PLA from January 2007 to December 2017. There were 58 males and 45 females, aged 28-69 years, with an average age of 43.8 years. Among the patients, 84 had normal sacrums while 19 had dysmorphic sacrums. The accuracy and placement time of two kinds of sacroiliac screw were compared by X-ray fluoroscopy (46 patients with normal sacroiliac screw placement and 11 with dysmorphic sacroiliac screw placement) or CT guidance (66 patients with normal sacroiliac screw placement and 18 with dysmorphic sacroiliac screw placement) were compared. Matta standard was used to evaluate the imageological healing and clinical prognosis 9 months after operation. Results: In normal sacrum group, there was no statistical difference in placement accuracy between X-ray fluoroscopy [89%(41/46)] and CT guided technique [94%(62/66)](P>0.05). In dysmorphic sacrum group, the placement accuracy of CT guided technique [89%(17/18)] was significantly higher than that of X-ray fluoroscopy [55%(6/11)] (P<0.05). The screw placement time of X-ray fluoroscopy [(39.1±4.9)minutes] was significantly shorter than that of CT guided technique (54.7±3.8)minutes for normal sacrums (P<0.05), and there was no significant difference in terms of placement time by CT guided technique for dysmorphic sacrums (P>0.05). Nine months after operation, in normal sacrum group, the X-ray fluoroscopy had the excellent rate of 74% (34/46), good rate of 26% (12/46) for healing and the excellent rate of 83% (38/46), good rate of 17% (8/46) for clinical prognosis, showing no significant difference from those of CT guided technique [83%(55/66), 17%(11/66); 92%(61/66), 8%(5/66)] (P>0.05). In dysmorphic sacrum group 9 months after operation, the excellent and good rate of X-ray fluoroscopy [excellent 9%(1/11), good 64%(7/11)] was significantly lower than that of CT guided technique in terms of imageological healing [excellent 56%(10/18), good 39%(7/18)](P<0.05), while no significant difference was found in terms of clinical prognosis [excellent 55%(6/11), good 36%(4/11) vs. excellent 78%(14/18), good 22%(4/18)](P>0.05). Conclusions X-ray fluoroscopy for the fixation of type Tile B or C posterior ring fractures or dislocations of the normal sacrum takes much shorter time, although the comparable effect with CT guidance in aspects of placement accuracy and bone healing rate. For the dysmorphic sacrum, CT guidance allows more accurate screw placement and has better healing rate than X-ray fluoroscopy.