Objective To investigate the mechanism of morin-induced autophagy in non-small cell lung cancer A549 cells based on mTOR/STAT3 signaling axis.Methods A549 cells were divided into blank group and 30,60,90,120 and 150 μg·mL-1 of morin groups.After 24,48 and 72 hours of culture,the cell proliferation activity was detected by CCK-8 method,and the cell inhibition rate was calculated.A549 cells were divided into blank group and 30,90,150 μg·mL-1 morin groups.After 14 days of culture,the cell proliferation was detected by colony formation assay.After 24 hours of culture,the cell proliferation ability was detected by BeyoClickTM EdU-488.Apoptosis was detected by flow cytometry;acridine orange staining was used to detect cell autophagy;the formation of autophagosomes was observed by transmission electron microscopy.Western Blot was used to detect the expression levels of apoptosis,autophagy and mTOR/STAT3 signaling axis-related proteins in cells.A549 cells were divided into blank group,blank group + chloroquine(10 μg·mL-1)group,morin(30,150 μg·mL-1)group,morin(30,150 μg·mL-1)+ chloroquine(10 μg·mL-1)group.After 48 hours of intervention,the cell activity was detected by CCK-8 method,and the cell survival rate was calculated.Results Compared with the blank group,the inhibition rate of A549 cells in 60,90,120,150 μ g·mL-1 of morin group was significantly increased after 24 hours of intervention(P＜0.05,P＜0.001).The inhibition rates of A549 cells in 30,60,90,120 and 150 μg·mL-1 of morin groups were significantly increased after 48 and 72 hours of intervention(P＜0.001).The number of A549 cell colonies and the number of green fluorescent proliferation positive cells in the 30,90,150 μg·mL-1 of morin groups were significantly decreased(P＜0.01,P＜0.001),the apoptosis rate was significantly increased(P＜0.01,P＜0.001),and the protein expression level of cleaved-PARP was significantly increased(P＜0.001).The protein expression levels of p-P38/P38 MAPK in A549 cells of 90 and 150 μg·mL-1 of morin groups were significantly increased(P＜0.01,P＜0.001).Different degrees of orange fluorescence appeared in A549 cells of 30,90 and 150 μg·mL-1 of morin groups,and the orange fluorescence of 90 and 150 μg·mL-1 of morin groups was significant.Autophagosomes and autolysosomes appeared in the cytoplasm of A549 cells in 150 μg·mL-1 of morin group,respectively.The protein expression of LC3-Ⅱ in A549 cells of 150 μg·mL-1 of morin group was significantly up-regulated(P＜0.05).The protein expression of Atg16L1-Ⅱ in A549 cells of 90,150 μg·mL-1 of morin group was significantly up-regulated(P＜0.001),and the protein expressions of p-mTOR/mTOR and p-STAT3/STAT3 were significantly down-regulated(P＜0.001).Compared with the morin(150 μg·mL-1)group,the survival rate of A549 cells in the morin(150 μg·mL-1)+chloroquine(10 μg·mL-1)group was significantly increased(P＜0.05).Conclusion Morin can promote the apoptosis of A549 cells and induce autophagy in A549 cells,and the mechanism may be related to mTOR/STAT3 axis.

Insomnia disorder is a common clinical mental disorder.Currently, clinical subtyping of insomnia disorder relies primarily on symptomatic descriptions, lacking objective measures and subtyping-based treatment approaches. In recent years, increasing attention has been drawn to sleep electroencephalography (EEG) as a valuable tool for observing abnormal sleep architecture and continuity of insomnia disorder. Sleep EEG analysis holds the potential to elucidate the underlying biological mechanisms of insomnia disorder, facilitating data-driven subtyping and enhancing personalized therapeutic strategies.Five types of sleep EEG subtypes of insomnia disorder were systematically searched and summarized: classifications derived from objective sleep duration, power spectral characteristics, cyclic alternating pattern, spindle and microarousal.EEG characteristics of each subtype and clinical outcomes are discussed.This review aims to provide evidence-based insights for clinical subtyping and personalized treatment of insomnia disorder.

Paradoxical insomnia(Para-I),also known as pseudoinsomnia or sleep state misperception,is a condition in which the patient complains of severe insomnia but has no objective evidence of sleep disorder,and daytime functioning may be disrupted disproportionately to the degree of patient-reported sleep loss.Para-I is characterized by overestimation of sleep latency(SL)and underestimation of total sleep time(TST).Incorrect assessment of sleep quality hinders the diagnosis,evaluation of severity,and assessment of clinical efficacy of sleep disorders.The pathogenesis of Para-I remains unclear,but may be related to factors such as depression,anxiety,personality traits,social relationships and specific changes in brain structure and function.Studies on the polysomnography(PSG)of the patients with insomnia have found that changes in non-rapid eye movement(NREM)and rapid eye movement(REM)sleep may be related to the degree of subjective-objective sleep discrepancy.PSG is a valuable diagnostic tool for sleep disorders.It allows for the analysis of sleep structure and related physiological and behavioral changes by monitoring various parameters,including electroencephalogram(EEG),electromyogram(EMG),electrooculogram(EOG),oro-nasal airflow,thoracic and abdominal respiratory motions,oxygen saturation,electrocardiogram(ECG)and snoring.In recent years,studies have increasingly explored the sleep EEG and treatment of Para-I with PSG,resulting in significant progress.This article reviews the latest advances in the electroencephalographic characteristics and treatment of Para-I,providing new ideas for precise treatment.

Cancer still has elevated morbidity and mortality, which undoubtedly impacts the life quality of affected individuals. Remarkable advances have been made in cancer therapy, although the toxicities of traditional therapies remain an obvious challenge. Dahuang Zhechong Pill (DHZCP), developed by Zhongjing Zhang in the Synopsis of the Golden Chamber, represents an effective anticancer traditional Chinese medicine (TCM). In this review, it was found that DHZCP is therapeutically utilized in liver, lung, gastric, pancreatic and other cancers in clinic. Pharmacological evidence showed that its anti-tumor mechanisms mainly involve induced cell cycle arrest, apoptosis and autophagy, as well as suppressed tumor cell proliferation, obstructed angiogenesis and metastasis, enhanced immunity, and reversal of multidrug resistance. The present review provides a solid basis for the clinical application of DHZCP and may promote the wide use of TCM in clinical antitumor application.

ObjectiveTo evaluate the effects of dehydrocostus lactone （DL） on the proliferation， apoptosis， and autophagy of human lung cancer cell A549 and to elucidate its related mechanism. MethodThe effect of DL with different concentrations （0， 5， 10， 15， 20， 25 μmol·L^-1） on the proliferation of human lung cancer A549 cells was investigated by cell counting kit-8 （CCK-8）， and its impact on the clonogenic ability of A549 cells was studied by cell clonogenic assay. The concentrations 10， 20 μmol·L^-1 were selected as DL low-dose group and high-dose group. Hoechst 33258 staining and western blot were used to observe the effect of DL on apoptosis of A549 cells. Autolysosomes were detected by acridine orange staining， and the expression level of microtubule-associated protein 1 light chain 3 （LC3） was determined by immunofluorescence and western blot. In addition， the effects of DL in combination with autophagy inhibitors bafilomycin A1 （BAF-A1） or 3-methyladenine （3-MA） on the autophagy of A549 cells was checked by CCK-8 assay. Finally， the role of DL in the regulation of A549 cell signaling pathway was explored by Western blot. ResultCompared with the conditions in the control group， the survival rate of A549 cells in the DL groups （10， 15， 20， 25 μmol·L^-1） was decreased （P<0.01）， and 5 μmol·L^-1 DL could inhibited the formation of A549 clone cells （P<0.01）， indicating that DL could inhibit the proliferation of human lung cancer A549 cells. The number of apoptotic cells was higher in both DL low-dose and high-dose groups than that in the control group， and the expression of apoptosis-related proteins poly （ADP ribose） polymerase （PARP） and B lymphocytoma-2 （Bcl-2）-associated X protein （Bax） were up-regulated （P<0.05， P<0.01）， while the expression of Bcl-2 was down-regulated （P<0.01） in DL high-dose group. The acridine orange staining showed that the orange fluorescence in the DL high-dose group was enhanced compared with that in the control group， indicating that DL could dramatically promote the formation of autolysosomes. Moreover， 20 μmol·L^-1 DL could increase the orange fluorescent particles of LC3 and up-regulated the expression level of LC3 Ⅱ （P<0.01）. After addition of autophagy inhibitors， the sensitivity of A549 cells to the effects of DL was attenuated （P<0.01）， which suggested that autophagy was involved in DL-induced A549 cell death. Compared with the control group， DL high-dose group had increased expression of autophagy-related protein 3 （Atg3） and autophagy-related protein 5 （Atg5） while reduced phosphorylation levels of protein kinase B （Akt）， mammalian target of rapamycin （mTOR） and signal transducer and activator of transcription 3 （STAT3） （P<0.05， P<0.01）. ConclusionDL could activate apoptosis and autophagy to inhibit the proliferation and clonogenic ability of A549 cells via suppressing Akt/mTOR/STAT3 signaling pathway.

BACKGROUND: Synovectomy has been introduced into total knee arthroplasty (TKA) with the aim of relieving pain and inflammation of the synovium. However, there are no long-term, comparative data to evaluate the effect of synovectomy in TKA. This study was aimed at assessing pain, function, and complications in patients undergoing synovectomy during TKA for osteoarthritis (OA) at long-term follow-up. METHODS: This was a prospective randomized controlled trial of 42 consecutive patients who underwent staged bilateral TKA. Patients undergoing the first-side TKA were allocated to receive TKA with or without synovectomy followed by a 3-month washout period and crossover to the other strategy for the opposite-side TKA. The overall efficacy of both strategies was evaluated by determination of blood loss, the Knee Society score (KSS), and knee inflammation conditions during a 3-month postoperative period. The postoperative pain, range of motion (ROM), and complications were sequentially evaluated to compare the two groups until 10 years after surgery. RESULTS: At the 10-year follow-up, both groups had a similarly significantly improved ROM (114.88 ± 9.84° vs. 114.02 ± 9.43°, t  = 0.221, P  = 0.815) and pain relief with no differences between the two groups (1.0 [1.0] vs. 1.0 [1.5], U  = 789.500, P  = 0.613). Similar changes in total blood loss, KSS, and knee inflammation were found in both groups during 3 months postoperatively ( P  > 0.05). Additionally, there was no significant difference regarding complications and satisfaction between the two groups ( P  > 0.05). CONCLUSIONS: Synovectomy in conjunction with TKA for primary OA does not seem to provide any benefit regarding postoperative pain, ROM, and satisfaction during a 10-year follow-up. In addition, it may not result in more blood loss and increased incidence of long-term complications. Based on our long-term findings, it should not be performed routinely. TRIAL REGISTRATION Chinese Clinical Trial Registry, ChiCTR-INR-16008245; https://www.chictr.org.cn/showproj.aspx?proj=13334 .
Humans ; Arthroplasty, Replacement, Knee/methods* ; Synovectomy/methods* ; Osteoarthritis, Knee/surgery* ; Prospective Studies ; Pain, Postoperative ; Inflammation/etiology* ; Range of Motion, Articular ; Knee Joint/surgery* ; Treatment Outcome ; Knee Prosthesis/adverse effects*

OBJECTIVE: To investigate the risk factors of contralateral anterior cruciate ligament (ACL) injury after primary ACL reconstruction. METHODS: A retrospective review was conducted on the 716 patients with ACL injury who received primary ACL reconstruction surgery and met the selection criteria between January 2012 and September 2018. After a mean follow-up period of 7.6 years (range, 4-10 years), 65 patients (9.1%) experienced contralateral ACL injury (injured group) and 651 patients (90.9%) did not (uninjured group). There was no significant difference in age, body mass index, and preoperative Lachman test degree between groups ( P>0.05). However, the proportion of female in the injured group was significantly higher than that of male ( P<0.05), and the preoperative posterior tibial slope (PTS) was significantly higher than that of the uninjured group ( P<0.05). Using the outcome of contralateral ACL injury as the dependent variable, the clinical data of the patient was first used as the independent variable, and univariate COX regression was used to analyze the prognostic influencing factors. Then, the indicators with differences in univariate COX regression were used as the independent variable, and multivariate COX regression was used to analyze the independent risk factors affecting prognosis. Log-Rank (Mantel-Cox) test was used to test and analyze the occurrence time of contralateral ACL injury in patients of different genders; X-tile software was used to analyze the occurrence time of contralateral ACL injury in patients with different PTS using Log-Rank (Mantel-Cox) test and PTS cut-off values. RESULTS: Univariate COX regression analysis showed that gender and PTS were influence factors for contralateral ACL injury ( P<0.05); further multivariate COX regression analysis showed that female and increased PTS were independent risk factors for contralateral ACL injury ( P<0.05). The Log-Rank (Mantel-Cox) test results showed that the contralateral ACL injury occurred in female at 8.853 (8.600, 9.106) years, which was significantly shorter than that in male [9.661 (9.503, 9.819) years] ( χ ²=20.323, P<0.001). Using X-tile software to analyze the cut-off value of PTS, it was found that the cut-off value of PTS for contralateral ACL injury was 10.92°. According to the Log-Rank (Mantel-Cox) test, it was found that the contralateral ACL injury occurred in 5.762 (4.981, 6.543) years in patients with PTS≥10.92°, which was significantly shorter than patients with PTS<10.92° [9.751 (9.650, 9.853) years]( χ ² =302.479, P<0.001). CONCLUSION Female and PTS≥10.92° after primary ACL reconstruction are independent risk factors for contralateral ACL injury.
Humans ; Male ; Female ; Anterior Cruciate Ligament Injuries/surgery* ; Knee Joint/surgery* ; Risk Factors ; Tibia/surgery* ; Retrospective Studies ; Anterior Cruciate Ligament Reconstruction/methods*

Objective:To investigate the feasibility of transperineal ultrasound in quantitative assessment of posterior compartment prolapse among Chinese women.Methods:The prospective multicenter study enrolled 485 women between January 2017 and January 2019. All patients underwent a standard clinical interview, pelvic organ prolapse quantification (POP-Q) examination and transperineal ultrasound examination. Volume data of transperineal ultrasound examinations were obtained at rest and in maximal Valsalva maneuver.Results:The higher POP-Q stage of posterior compartment, the lower rectal ampulla position in maximal Valsalva maneuver (POP-Q stage=0 vs POP-Q stage=1, P<0.001; POP-Q stage=1 vs POP-Q stage≥2, P<0.001), and the greater rectal ampulla hypermobility (POP-Q stage=0 vs POP-Q stage=1, P<0.001; POP-Q stage=1 vs POP-Q stage≥ 2, P=0.007). The rectal ampulla position at rest and in maximal Valsalva maneuver and rectocele depth were correlated with prolapse symptoms ( r=-0.200, P<0.001; r=-0.252, P<0.001; r=0.086, P=0.045). The corresponding cut-off values of rectal ampulla position in maximal Valsalva in diagnosing posterior compartment prolapse (POP-Q stage ≥1) and clinical significant posterior compartment prolapse (POP-Q stage ≥2) were 7.32 mm below the symphysis pubis and 12 mm below the symphysis pubis, respectively, with the area under the ROC curve as 0.75 and 0.85, respectively. Conclusions:The ultrasonic measurements by transperineal ultrasound is significantly associated with POP-Q examination in posterior compartment, and it is demonstrated as a useful tool in quantitative assessment of the severity of posterior compartment prolapse.

Objective:To evaluate the therapeutic effect and safety of microwave ablation plus curettage.Methods:Clinical data of a single group of 12 patients admitted to our hospital from June 2015 to June 2019 who underwent microwave ablation for bone metastasis were retrospectively analyzed. There were 7 males and 5 females, with an average age of 59.5±10.1 (range from 43 to 79) years old. Curettage plus bone cement after microwave ablation was performedand when the lesion involved the sacroiliac joint surface and impaired the bearing arch, the internal fixation was performed with plate and screws. 8 among the 12 patients underwent curettage, bone cement and metal internal fixation after microwave ablation. After microwave ablation, and 4 patients underwent curettage and reconstruction with bone cement alone. The patients were followed up aftersurgery. The observational indicators included: patient survival rate, local recurrence rate, perioperative complications, Operation time, intraoperative blood loss, VAS score, SF-36 score, and MSTS function score. VAS scores and SF-36 scores were recorded before surgery, 1 week after surgery and 3 months after surgery.Results:12 patients were followed up for 4-27 months, with an average follow-up time of 14.2 months and a median follow-up time of 9 months. During the follow-up, 7 patients died and 5 survived, with a 6-month survival rate of 81.8%, a 1-year survival rate of 40.9% and a 2-year survival rate of 27.3%. The mean survival time of the dying patients was 8.8 months (5-18 months), and the median survival time was 7 months. Operation time was 101±21 min and intraoperative blood loss was 295±108 ml. During the follow-up period, local recurrence rate was 8.3% (1/12), and the average postoperative time to recurrence was 4 months. There were no complications such as wound nonunion, wound infection, deep infection, vascular nerve heat injury and internal fixation failure. VAS score before surgery was 7.5 (7, 8) points, VAS score 1 week after surgery was 5 (4, 5) points, and VAS score 3 months after surgery was 5 (4, 5.75) points ( P<0.05). Preoperative SF-36 score was 23 (21, 24.75) points, 1 week after surgery SF-36 score was 47 (46, 54) points, and 3 months after surgery SF-36 score was 50.5 (47, 55.25) points ( P<0.05). 3 months after the operation, the MSTS score was 20.5 (16.25, 21.75) points, of which excellent 16.7% (2/12), good 50% (6/12), medium 16.7% (2/12), poor 16.7% (2/12), and the overall excellent and good rate was 66.7% (8/12). Conclusion:Microwave ablation plus curettage for pelvic metastasis of malignant tumor can significantly relieve local pain, rebuild limb function and improve patients' quality of life, and it`s an alternative treatment for pelvic metastases.