To explore the inhibitory effect of ginkgolide B (GB) on MH7A human fibroblast-like synoviocytes (FLS) and its potential mechanism. Firstly, 20 μg/L tumor necrosis factor-α (TNF-α) was pretreated with MH7A to establish a cell model of arthritis. After incubation of MH7A cells with various concentrations of GB, CCK-8 assay, Transwell assay, and flow cytometry (FCM) were separately used to detect cell viability, cell invasion, and cell apoptosis rate and cell cycle; Real-time quantitative PCR and Western blot assay were performed to detect the apoptosis- and cycle-related gene transcriptions and protein expressions, respectively. The results showed that compared with the control group, GB dose- and time-dependently suppressed cell viability to a greater extent; GB significantly reduced cell invasive ability and increased cell apoptosis rate and proportion of G0/G1 phase in MH7A cells, along with increased transcription levels of Bcl-2-associated X protein (Bax) and p21 mRNA and decreased transcription levels of Bcl-2, myeloid cell leukemia 1(Mcl-1), protein kinase B (PKB; AKT), IP3K, Cyclin D1 and cyclin-dependent kinase 4 (CDK4) mRNA; GB remarkably increased expression levels of Bax, p21, and cleaved-Caspase 3 protein and decreased expression levels of Bcl-2, Mcl-1, p-AKT, p-PI3K, Cyclin D1, and CDK4 protein, with decreased ratios of p-PI3K/PI3K, p-AKT/AKT, and Bcl-2/Bax. In conclusion, GB blocks the G1-to-S cell cycle transition, suppresses cell viability and cell invasion and induces cell apoptosis of MH7A human RA-FLS via suppressing the PI3K/AKT signaling pathway.

Objective To explore the clinical efficacy of swallowing training combined with Vitalstim neuromuscular electrical stimulation in patients with dysphagia after stroke for providing a scientific basis for improving the swallowing function.Methods A total of 100 patients with post-stroke dysphagia were enrolled and randomly divided into control group(n=50,conventional swallowing training)and observation group(n=50,conventional swallowing training+Vitalstim neuromuscular electrical stimulation),with a total treatment course of 3 weeks.The swallowing functions before and after treatment was compared between two groups.Results After treatment,observation group scored lower on the water swallow test and the Rosenbek Penetration-Aspiration Scale(P<0.05),while higher on the Functional Oral Intake Scale and the Functional Independence Measure as compared with control group(P<0.05).In addition,the score of Nutritional Risk Screening 2002(NRS2002)was higher in observation group than in control group(P<0.05).Conclusion The combination of swallowing training and Vitalstim neuromuscular electrical stimulator can effectively improve the swallowing function of stroke patients and has high clinical application value,worthy of further promotion and research.

Resistant ovary syndrome(ROS)and premature ovarian insufficiency(POI)fall under the cat-egory of hypogonadotropic amenorrhea,sharing similar clinical features that often pose challenges in differentiation.ROS can be easily misdiagnosed as POI,which presents a significant obstacle to subsequent treatment.Therefore,it is crucial for patients with fertility requirements to have a clear understanding of the etiology,clinical features,and diagnostic criteria of ROS and POI in order to establish an early diagnosis and develop an appropriate treatment plan.This article provides a systematic and comprehensive discussion on the research progress regarding the eti-ology and pathogenesis,clinical features and diagnosis,as well as individualized management of ROS and POI.The aim is to offer reference for clinicians in achieving early clarification of diagnoses,avoiding misdiagnosis or mistreatment,while assisting patients in improving symptoms and realizing their fertility aspirations through person-alized management.

Objective To investigate changes in the pro-inflammatory mediator S100A8/9 and NLRP3/Caspase-1/IL-1β pathway in a rat kidney-aging model induced by D-galactose.Methods Twelve SD rats were divided into control and D-galactose groups,and injected subcutaneously in the back of the neck with D-galactose(150 mg/kg)to establish a rat model of kidney aging.Kidney samples were collected under anesthesia after 8 weeks.Kidneys were stained for senescence-associated beta-galactosidase(SA-β-Gal),mRNA expression levels of the aging-related genes p21,p16,and p53 were detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR),and histopathological changes were observed by hematoxylin-eosin(HE)and Masson staining.Serum urea nitrogen and creatinine,and catalase(CAT),glutathione peroxidase(GSH-PX),superoxide dismutase(SOD),and malondialdehyde(MDA)levels in the kidney tissues were detected.Reactive oxygen species(ROS)were detected by dihydroethdium staining and protein expression levels of collagen Ⅲ,α-smooth muscle actin(α-SMA),Protein expression of S100A8/9 was detected by immunofluorescence,and transforming growth factor(TGF)-β1 levels in kidney tissues and key factors in the NLRP3/Caspase-1/IL-1β inflammatory pathway were detected by Western Blot.A renal senescence model using HK-2 cells was constructed using H2O2 in vitro,and expression levels of the senescence proteins p21 and p16 and mRNA expression levels of the inflammatory factors IL-18 and tumor necrosis factor-α(TNF-α)were detected.Cell senescence was observed by SA-β-Gal staining.The effects of the S100A8/9 inhibitor paquinimod on expression levels of S100A8/9 and NLRP3/Caspase-1/IL-1β pathway-related proteins in the aging model were also detected.Results mRNA levels of the aging genes p21,p16,and p53 in kidney tissues were significantly increased in rats in the D-galactose group compared with the control group(P＜0.01),and SA-β-Gal staining showed a significant increase in senescent cells(P＜0.01).Serum blood urea nitrogen and creatinine levels increased(P＜0.05),CAT,GSH-PX,and SOD activities decreased(P＜0.01),while MDA activity increased in the D-galactose group(P＜0.01).Collagen Ⅲ,α-SMA,and TGFβ1 expression and the ROS content in tissues increased(P＜0.05).Glomeruli were atrophied or absent in the D-galactose group,the lumens of the renal sacs and renal tubules were enlarged,the nuclei were deeply stained and constricted,and numerous collagen fibers were deposited.Levels of S100A8 and S100A9 protein(P＜0.01),as well as NLRP3,Caspase-1,and IL-1β increased(P＜0.05).Paquinimod alleviated HK-2 cell senescence and decreased expression levels of the senescence proteins p21 and p16,and mRNA levels of the inflammatory factors IL-18 and TNF-α(P＜0.05,P＜0.01).The number of senile cells was also decreased,shown by SA-β-Gal staining(P＜0.01).Paquinimod also inhibited the protein expression of S100A8 and S100A9(P＜0.01)and NLRP3,Caspase-1,and IL-1β(P＜0.05 or P＜0.01).Conclusions S100A8/9 participates in the chronic inflammatory response by activating the NLRP3/Caspase-1/IL-1β pathway,thereby promoting D-galactose-induced renal aging.

Objective To explore the clinical efficacy of swallowing training combined with Vitalstim neuromuscular electrical stimulation in patients with dysphagia after stroke for providing a scientific basis for improving the swallowing function.Methods A total of 100 patients with post-stroke dysphagia were enrolled and randomly divided into control group(n=50,conventional swallowing training)and observation group(n=50,conventional swallowing training+Vitalstim neuromuscular electrical stimulation),with a total treatment course of 3 weeks.The swallowing functions before and after treatment was compared between two groups.Results After treatment,observation group scored lower on the water swallow test and the Rosenbek Penetration-Aspiration Scale(P<0.05),while higher on the Functional Oral Intake Scale and the Functional Independence Measure as compared with control group(P<0.05).In addition,the score of Nutritional Risk Screening 2002(NRS2002)was higher in observation group than in control group(P<0.05).Conclusion The combination of swallowing training and Vitalstim neuromuscular electrical stimulator can effectively improve the swallowing function of stroke patients and has high clinical application value,worthy of further promotion and research.

Resistant ovary syndrome(ROS)and premature ovarian insufficiency(POI)fall under the cat-egory of hypogonadotropic amenorrhea,sharing similar clinical features that often pose challenges in differentiation.ROS can be easily misdiagnosed as POI,which presents a significant obstacle to subsequent treatment.Therefore,it is crucial for patients with fertility requirements to have a clear understanding of the etiology,clinical features,and diagnostic criteria of ROS and POI in order to establish an early diagnosis and develop an appropriate treatment plan.This article provides a systematic and comprehensive discussion on the research progress regarding the eti-ology and pathogenesis,clinical features and diagnosis,as well as individualized management of ROS and POI.The aim is to offer reference for clinicians in achieving early clarification of diagnoses,avoiding misdiagnosis or mistreatment,while assisting patients in improving symptoms and realizing their fertility aspirations through person-alized management.

Objective To investigate changes in the pro-inflammatory mediator S100A8/9 and NLRP3/Caspase-1/IL-1β pathway in a rat kidney-aging model induced by D-galactose.Methods Twelve SD rats were divided into control and D-galactose groups,and injected subcutaneously in the back of the neck with D-galactose(150 mg/kg)to establish a rat model of kidney aging.Kidney samples were collected under anesthesia after 8 weeks.Kidneys were stained for senescence-associated beta-galactosidase(SA-β-Gal),mRNA expression levels of the aging-related genes p21,p16,and p53 were detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR),and histopathological changes were observed by hematoxylin-eosin(HE)and Masson staining.Serum urea nitrogen and creatinine,and catalase(CAT),glutathione peroxidase(GSH-PX),superoxide dismutase(SOD),and malondialdehyde(MDA)levels in the kidney tissues were detected.Reactive oxygen species(ROS)were detected by dihydroethdium staining and protein expression levels of collagen Ⅲ,α-smooth muscle actin(α-SMA),Protein expression of S100A8/9 was detected by immunofluorescence,and transforming growth factor(TGF)-β1 levels in kidney tissues and key factors in the NLRP3/Caspase-1/IL-1β inflammatory pathway were detected by Western Blot.A renal senescence model using HK-2 cells was constructed using H2O2 in vitro,and expression levels of the senescence proteins p21 and p16 and mRNA expression levels of the inflammatory factors IL-18 and tumor necrosis factor-α(TNF-α)were detected.Cell senescence was observed by SA-β-Gal staining.The effects of the S100A8/9 inhibitor paquinimod on expression levels of S100A8/9 and NLRP3/Caspase-1/IL-1β pathway-related proteins in the aging model were also detected.Results mRNA levels of the aging genes p21,p16,and p53 in kidney tissues were significantly increased in rats in the D-galactose group compared with the control group(P＜0.01),and SA-β-Gal staining showed a significant increase in senescent cells(P＜0.01).Serum blood urea nitrogen and creatinine levels increased(P＜0.05),CAT,GSH-PX,and SOD activities decreased(P＜0.01),while MDA activity increased in the D-galactose group(P＜0.01).Collagen Ⅲ,α-SMA,and TGFβ1 expression and the ROS content in tissues increased(P＜0.05).Glomeruli were atrophied or absent in the D-galactose group,the lumens of the renal sacs and renal tubules were enlarged,the nuclei were deeply stained and constricted,and numerous collagen fibers were deposited.Levels of S100A8 and S100A9 protein(P＜0.01),as well as NLRP3,Caspase-1,and IL-1β increased(P＜0.05).Paquinimod alleviated HK-2 cell senescence and decreased expression levels of the senescence proteins p21 and p16,and mRNA levels of the inflammatory factors IL-18 and TNF-α(P＜0.05,P＜0.01).The number of senile cells was also decreased,shown by SA-β-Gal staining(P＜0.01).Paquinimod also inhibited the protein expression of S100A8 and S100A9(P＜0.01)and NLRP3,Caspase-1,and IL-1β(P＜0.05 or P＜0.01).Conclusions S100A8/9 participates in the chronic inflammatory response by activating the NLRP3/Caspase-1/IL-1β pathway,thereby promoting D-galactose-induced renal aging.

Objective To explore the main factors affecting the MV imager projection offset in the machine performance check(MPC)for Varian Vital Beam linear accelerator.Methods The MV imager projection offsets in the MPC after repairing the MV imaging arm encoder of shoulder motor,locking the treatment couch,and isocenter calibration were analyzed.Results MPC results revealed that the MV imager projection offset after repairing the MV imaging arm encoder of shoulder motor was(0.310±0.001)mm,significantly less than(0.450±0.010)mm in the blank group.The difference in MV imager projection offset between the isocenter calibration group and the blank group was trivial.The MV imager projection offset after locking the treatment couch was(0.240±0.030)mm,significantly less than(0.450±0.010)mm in the blank group.When MPC was carried out after repairing the imaging arm encoder and performing isocenter calibration,there was no significant statistical difference in MV imager center offset between the locked and unlocked treatment couch.Conclusion The damage of MV imaging arm encoder of shoulder motor is the main factor causing abnormal MV imager projection offsets.Locking the treatment couch before the MV imaging center check can reduce the results,but it cannot eliminate the MV imager projection offset.

N6-Methyladenosine (m⁶A) is the most abundant internal modification in eukaryotic mRNA, playing critical role in various bioprocesses. Like other epigenetic modifications, m⁶A modification can be catalyzed by the methyltransferase complex and erased dynamically to maintain cells homeostasis. Up to now, only two m⁶A demethylases have been reported, fat mass and obesity-associated protein (FTO) and alkylation protein AlkB homolog 5 (ALKBH5), involving in a wide range of mRNA biological progress, including mRNA shearing, export, metabolism and stability. Furthermore, they participate in many significantly biological signaling pathway, and contribute to the progress and development of cancer along with other diseases. In this review, we focus on the studies about structure, inhibitors development and biological function of FTO and ALKBH5.

Objective:To investigate the accuracy of pure titanium and cobalt-chromium alloy frameworks fabricated using the additive manufacturing (AM) of selective laser melting technology (SLM) for the mandibular implant-supported fixed prostheses and the maxillary removable partial denture (RPD), and to provide a reference for clinical application of SLM pure titanium frameworks.Methods:One edentulous mandibular model with implants and screw fixed abutments at bilateral canines and the first molars was selected and used as the mandibular full arch implant-supported model. At the same time, a Kennedy class Ⅰ maxillary dentition defect model was selected. The digital models were obtained by scanning the dental models, and the metal frameworks of the mandibular full arch implant-supported denture and the maxillary RPD (design model) were designed using the 3 Shape software. Meanwhile, 12 mandibular frameworks in the cobalt-chromium alloy and the pure titanium (6 in each group were treated with heat treatment, while the other 6 were not treated), and 7 maxillary frameworks in the cobalt-chromium alloy and the pure titanium were respectively made by SLM with the improved dual-laser metal printer. The axial direction of the printing powder accumulation was taken as the Z-axis. During the design process, the software (3Shape Dental System 2018) automatically generated the X-axis and Y-axis, X axis was the sagittal axis of the dental model and Y axis was the coronal axis of the dental model. The deviation of the interface center of the abutment of the digital model of the mandibular frameworks from the design model in the X, Y and Z axes was analyzed. As for the trueness of the mandibular framework, the larger the deviation data was, the worse the trueness was. The deviation of the whole maxillary framework and 7 measuring points (palatal plate center point and bilateral occlusal rests, I bars, proximal plates) were analyzed. The fitness of the whole maxillary framework to the design model was expressed by root mean square (RMS) of the deviation data, and the fitness of measuring points was expressed by the mean±standard deviation of the data. The trueness differences of each group before and after heat treatment of the mandibular framework and the fitness of the maxillary framework were compared.Results:The cobalt-chromium alloy frameworks showed lower trueness on the X, Y, Z-axes [(96.3±12.1), (86.3±11.4), (61.2±13.2) μm] than did the pure Ti frameworks [(82.3±11.2), (72.2±10.2), (51.2±11.6) μm] by SLM, and the heat treatment could reduce the discrepancy between the SLM frameworks and STL models, for pure titanium frameworks [(62.4±11.3), (55.2±13.2), (41.3±10.8) μm] and for cobalt-chromium alloy [(84.5±10.5), (72.3±11.2), (54.2±11.6) μm]. For the thin RPD major frameworks, pure titanium had better fitness [(121.3±17.0) μm] than cobalt-chromium alloy [(174.0±18.3) μm] by SLM, and the difference was statistically significant ( P<0.05). Conclusions:Pure titanium frameworks fabricated by SLM additive manufacturing technology exhibited better fitness and trueness than did the Co-Cr frameworks after heat treatment respectively, and this satisfied the requirements of implant-supported fixed prostheses and RPD major metal frameworks.