Breast cancer remains a leading cause of mortality in women worldwide.Triple-negative breast cancer(TNBC)is a particularly aggressive subtype characterized by rapid progression,poor prognosis,and lack of clear therapeutic targets.In the clinic,delineation of tumor heterogeneity and development of effective drugs continue to pose considerable challenges.Within the scope of our study,high hetero-geneity inherent to breast cancer was uncovered based on the landscape constructed from both tumor and healthy breast tissue samples.Notably,TNBC exhibited significant specificity regarding cell prolif-eration,differentiation,and disease progression.Significant associations between tumor grade,prog-nosis,and TNBC oncogenes were established via pseudotime trajectory analysis.Consequently,we further performed comprehensive characterization of the TNBC microenvironment.A crucial epithelial subcluster,E8,was identified as highly malignant and strongly associated with tumor cell proliferation in TNBC.Additionally,epithelial-mesenchymal transition(EMT)-associated fibroblast and M2 macrophage subclusters exerted an influence on E8 through cellular interactions,contributing to tumor growth.Characteristic genes in these three cluster cells could therefore serve as potential therapeutic targets for TNBC.The collective findings provided valuable insights that assisted in the screening of a series of therapeutic drugs,such as pelitinib.We further confirmed the anti-cancer effect of pelitinib in an orthotopic 4T1 tumor-bearing mouse model.Overall,our study sheds light on the unique characteristics of TNBC at single-cell resolution and the crucial cell types associated with tumor cell proliferation that may serve as potent tools in the development of effective anti-cancer drugs.

Objective:Three-dimensional simulation modeling technology was used to conduct three-dimensional reconstruction, classification and measurement for the anatomic structures of colorectal canal, tumor and key blood vessels in patients with rectal cancer before operation. And the accuracy of the data and information obtained for the prediction of anastomotic tension, important types of vascular variant, positioning of anatomical landmarks, etc. in laparoscopic-assisted radical resection of rectal cancer, and the guiding effect of operation was evaluated.Methods:The clinical data of 50 patients with laparoscopic-assisted radical resection of rectal cancer treated in the General Surgery Department, Beijing Ditan Hospital Capital Medical University from January 2019 to February 2021 were analyzed retrospectively, including 31 males and 19 females, aged from 42 to 83 years old, with an average age of (62.72 ±15.21) years. The patients were divided into two groups according to whether the patients underwent three-dimensional simulation reconstruction before operation. The patients who underwent abdomen pelvic enhancement CT and further three-dimensional reconstruction before surgery were taken as reconstruction group ( n=24), and the patients who were only routinely performed abdomen pelvic enhancement CT before operation were taken as control group ( n=26). For the patients in the reconstruction group, the CT images were modeled by Mimics software before operation, and the key data such as the length of colorectal and tumor, the correlation length of rigid structure of pelvic wall, the length of inferior mesenteric artery (IMA) from the bifurcation point of left and right arteria iliaca communis, the type and proportion of IMA variation, the length of left colonic artery (LCA) from the beginning of IMA and the distance between LCA and IMV were measured, and the consistency correlation coefficient (CCC) was analyzed with the actual data obtained during operation. And then the accuracy of three-dimensional simulation modeling technology for surgical guidance was evaluated. MedCalc 19.0 software was used for statistical analysis. Results:In the reconstruction group, regarding the data of each dimension of the model (intestinal tract, pelvic cavity, blood vessels) and the corresponding structural measurements during the operation, the consistent correlation coefficient (CCC) evaluation was more than 0.9. One case was predicted to have free splenic flexure of colon and one case actually had free splenic flexure of colon. The prediction accuracy was 100%. The IMA variants in the reconstruction group were divided into 4 types, all of which were verified by operation. Compared with the control group, the operation time ( P=0.011) and the location time (IMA, P=0.043; LCA, P=0.007; IMV, P=0.034) of each vessel in the reconstruction group were shorter, and the amount of intraoperative blood loss was less ( P=0.017). Conclusion:The application of three-dimensional simulation modeling technology before operation is helpful for the operator to accurately predict the intraoperative anastomotic tension, the type of IMA variation and the related diameter length, based on which the accurate operation plan can be made to guide the operation.

ObjectiveTo confirm the clinical efficacy and safety of Yishen Yangxin Anshen tablets in the treatment of insomnia （heart-blood deficiency and kidney-essence insufficiency syndrome）. MethodA randomized block， double-blind， placebo-controlled， multi-center clinical trial design method was adopted， and a total of 480 patients with insomnia due to deficiency of heart blood and insufficiency of kidney essence （treatment group-control group 3∶1） from seven hospitals （Guang'anmen Hospital， China Academy of Chinese Medical Sciences， The First Clinical Hospital， Jilin Province Academy of Traditional Chinese Medicine（TCM）， The Second Affiliated Hospital of Liaoning University of TCM， The First Affiliated Hospital of Henan University of Chinese Medicine， Henan Province Hospital of TCM， Hebei General Hospital， The First Hospital of Hunan University of Chinese Medicine） were enrolled. The treatment group was given Yishen Yangxin Anshen tablets and the control group received placebo tablets （4 tablets/time， 3 times/day， 4 weeks of administration， 4 weeks of follow-up after drug withdrawal）. The sleep dysfunction rating scale （SDRS） score， pittsburgh sleep quality index （PSQI） score， TCM， polysomnography （PSG） indicators from four hospital （Guang'anmen Hospital， China Academy of Chinese Medical Sciences， Henan Province Hospital of TCM， Hebei General Hospital， The First Hospital of Hunan University of Chinese Medicine）， and other efficacy indicators were compared between the two groups before and after treatment. Through general physical examination， laboratory examination， and observation of adverse events， the safety of the drugs was evaluated. ResultThe baseline indexes of the two groups showed no significant difference and thus the two groups were comparable. After treatment， the total score of SDRS in the treatment group was lower than that in the control group （P<0.01）. After drug withdrawal for 4 weeks， the total score of SDRS demonstrated no significant change in the treatment group as compared with that at the end of treatment， indicating that the rebound change of curative effect was not obvious. After treatment， the total score of PSQI in the treatment group decreased as compared with that in the control group （P<0.01）， and the change of total score of PSQI in the treatment group was statistically significant （P<0.05） after drug withdrawal for 4 weeks but small， indicating that the rebound change of curative effect was not obvious. After treatment， the total effective rate about the TCM symptoms in the treatment group was higher than that in the control group （χ²=137.521，P<0.01）. After treatment， the disappearance rates of single indexes in the treatment group， such as difficulty in falling asleep， easily waking up after sleeping， early awakening， short sleep time， dreamfulness， palpitation， forgetfulness， dizziness， mental fatigue， and weakness of waist and knee， increased compared with those in the control group （P<0.01）. After treatment， the treatment group demonstrated fewer awaking times （AT）， longer total sleep time （TST）， lower ATA/TST ratio， and higher sleep efficiency （%） than the control group （P<0.05）. No abnormal value or aggravation related to drugs was observed in either group. The incidence of adverse events in the treatment group and the control group was 5.57% and 8.40% respectively. No serious adverse events or adverse events leading to withdrawal happened in either group. ConclusionYishen Yangxin Anshen tablets is effective and safe for patients with insomnia of deficiency of heart-blood and insufficiency of kidney-essence.

Objective:To investigate the role and molecular mechanism of death receptor 5 (DR5) in early brain injury (EBI) after subarachnoid hemorrhage (SAH), as well as the neuroprotective effect of soluble DR5 (sDR5) on SAH.Methods:Experiment 1: SD rats were randomly divided into sham-operated group ( n=6) and SAH group (SAH model was established by carotid artery puncture, n=30), and the SAH group was further subdivided into post-SAH (6 h) group, post-SAH (12 h) group, post-SAH (24 h) group, post-SAH (48 h) group and post-SAH (72 h) group ( n=6); Western blotting was used to detect the expressions of tumor necrosis factor (TNF)-α and DR5; immunofluorescent DR5 and neuronal nuclear antigen (NeuN) double staining was used to evaluate the DR5 expression in neurons. Experiment 2: SD rats were randomly divided into sham-operated group, SAH group, Trail group (injected Trail agonist dordaviprone), and Trail+sDR5 group (injected dordaviprone+sDR5, n=6); at the 24 th h of successfully constructed SAH model, the caspase family protein levels were detected by Western blotting, and Tunel staining and immunofluorescent DR5 and Caspase-3 double staining were performed. Experiment 3: SD rats were divided into sham-operated group, SAH group, Trail group and Trail+sDR5 group ( n=6); long-term motor functions, by modified Gracia score, forelimb placement experiment, rotarod test and misstep experiment, were evaluated 5, 7 and 12 d after successfully constructed SAH model; and long-term learning and memory functions were detected by water maze experiment 14, 16, 18, 20 and 21 d after successfully constructed SAH model. Results:(1) Result of Experiment 1: the expressions of TNF-α and DR5 in sham-operated group, post-SAH (6 h) group, post-SAH (12 h) group, post-SAH (24 h) group, post-SAH (48 h) group and post-SAH (72 h) group were statistically different ( F=837.992, P<0.001; F=503.942, P<0.001), and these expressions peaked 24 h after SAH; immunofluorescent DR5 and NeuN double staining showed that DR5 was located in neurons after SAH. (2) Result of Experiment 2: compared with the SAH group and Trail group, the Trail+sDR5 group had significantly decreased levels of activated caspase-8, tBid and activated caspase-3, significantly decreased numbers of Tunel positive cells and DR5 and activated caspase-3 co-marked positive cells ( P<0.05). (3) Result of Experiment 3: compared with the SAH group and Trail group, the Trail+sDR5 group had significantly increased Garcia scores, decreased failure rate in forelimb placement experiment, prolonged duration of stick rotation, and decreased foot fault rate ( P<0.05), suggesting that sDR5 could improve the long-term motor function deficit after SAH; water maze experiment showed that 21 d after SAH, compared with the SAH group and Trail group, Trail+sDR5 group had significantly increased proportion of escape time in the original platform quadrat in total escape time and increased proportion of movement path in the original platform quadrat in total movement path after platform removal ( P<0.05), suggesting that sDR5 could improve long-term learning and memory impairment after SAH. Conclusion:The sDR5 can inhibit DR5-Trail-mediated neuronal apoptosis and improve long-term neurological functional deficits after SAH.

Objective:To investigate the correlation between 25 hydroxyvitamin D[25 (OH) D] level and sleep monitoring index in patients with severe altitude obstructive sleep apnea hypopnea syndrome (OSAHS).Methods:Sixty-six patients with severe OSAHS (AHI≥30 times/hour) diagnosed by apnea hypopnea index (AHI) who had lived at high altitude (1 800-4 193 m) for≥1 year were included in the experimental group. The patients underwent polysomnography monitoring in Sleep Medicine Center of Qinghai Red Cross Hospital from June to December 2021. In addition, healthy volunteers matched the experimental group by gender, age, ethnicity and living altitude during the same period were selected for polysomnography monitoring. Finally, 48 healthy volunteers with AHI<5 times/hour were included as the control group. 25(OH)D level and its deficiency were compared between the two groups. The experimental group was further divided into severe deficiency group [25(OH)D≤10 μg/L], the deficiency group [10 μg/L<25(OH)D≤20 μg/L] and the non-deficiency group [25(OH)D>20 μg/L] according to 25(OH)D level, and the differences of sleep parameters among the three groups were compared. Correlation analysis and multifactor linear regression analysis were performed on the factors that may affect the level of 25(OH)D in patients with severe OSAHS.Results:A total of 114 adults living on the plateau for at least one year were enrolled, including 66 in the experimental group and 48 in the control group. 25(OH)D deficiency (≤30 μg/L) was found in all the individuals included in the experimental group and the control group, and the 25(OH)D level of the two groups was [(13.13±4.05) vs (13.68±4.60) μg/L, P=0.507] and there was no significant difference in deficiency degree (all P>0.05). Within the experimental group, rapid eye movement (REM) sleep time and proportion (REM%) and sleep awakening time of 25(OH)D non-deficiency group, were significantly lower than those in severe deficiency group (all P<0.05), and sleep efficiency in 25(OH)D non-deficiency group was significantly higher than that in severe deficiency group and deficiency group (all P<0.05). Spearman correlation analysis showed that the level of 25 (OH) D in experimental group were positively correlated with serum calcium ion level ( r=0.293, P=0.017) and sleep efficiency ( r=0.309, P=0.011), and were negatively correlated with age ( r=-0.298, P=0.015), REM sleep time ( r=-0.401, P=0.001), REM% ( r=-0.421, P<0.001) and awakening time ( r=-0.362, P=0.003). Multifactor linear regression analysis suggested that serum calcium, REM sleep time and history of hypertension were the predictors of 25(OH)D level in severe OSAHS at high altitude. Conclusions:There is a correlation between sleep monitoring indexes and serum 25(OH)D level in patients with severe OSAHS at high altitude. The longer the REM sleep time, the lower the 25(OH)D level. Meanwhile, there is a certain relationship between co-morbidity hypertension and 25(OH)D level in patients with severe OSAHS at high altitude.

Objective:To explore the classification and treatment strategy of femoral neck fracture combined with ipsilateral intertrochanteric fracture.Methods:A retrospective analysis was performed of the 44 patients who had been admitted to Department of Orthopedics, Liangxiang Teaching Hospital, Capital Medical University from March 2003 to March 2019 for femoral neck fracture combined with ipsilateral intertrochanteric fracture. They were 19 males and 25 females, aged from 37 to 93 years (average, 77.9 years). According to the anatomical location and displacement severity, the femoral neck fractures were divided into 3 types while the intertrochanteric fractures were classified as stable or unstable ones. There were 3 cases of type Ⅰ which were completely extracapsular ones, 31 cases of type Ⅱ which were intracapsular stable ones, and 10 cases of type Ⅲ which were intracapsular unstable ones. Types Ⅰ and Ⅱ fractures were treated with intramedullary fixation, and type Ⅲ fractures with cemented hip hemi-replacement+reduction and fixation of the intertrochanteric fracture with Kirschner wires and steel cables. Recorded were fracture healing time, function of the affected hip and complications.Results:The 44 patients were followed up for at least 2 years. The fracture healing time for the 3 patients with type I fracture averaged 5.6 months (from 4.4 to 6.8 months); their hip function at the last follow-up, evaluated by the Harris scoring system, was excellent in 2 cases and good in one case. For 30 of the 31 patients with type Ⅱ fracture, the fracture healing time averaged 7.2 months (from 5.1 to 9.3 months); their hip function at the last follow-up, evaluated by the Harris scoring system, was excellent in 18 cases, good in 6 cases, fair in 5 cases, and poor in 2 cases, giving an excellent and good rate of 77.4% (24/31). As for complications, withdrawal or cutting-out of the head screw happened in 6 cases, infection in one case and nonunion in one case. In 10 patients with type Ⅲ fracture, the hip function at the last follow-up was excellent in 7 cases, good in 2 cases and poor in one case.Conclusions:For type Ⅰ and type Ⅱ femoral neck fractures combined with ipsilateral intertrochanteric fracture, intramedullary fixation with angulation stability may be a proper choice. For type Ⅲ ones, hip joint replacement should be the first choice.

Objective:To analyze the prevalence and distribution characteristics of diffuse idiopathic skeletal hyperostosis (DISH) in hospitalized patients with thoracic ossification of the ligamentum flavum (TOLF).Methods:The clinical records of 132 consecutive TOLF patients from January 2018 to June 2019 were retrospectively studied. DISH was identified by the preoperative X-ray and CT and its prevalence was calculated. The prevalence of patients with different genders, different age groups and different ossification types was compared. The segmental distribution of DISH and the distribution in the upper (T 1-T 4), middle (T 5-T 8), and lower thoracic spine (T 9-T 12) were analyzed. Ossification degree of DISH was evaluated based on the Meta scoring system. The demographic characteristics (age, gender, BMI, etc.) were compared between DISH and non-DISH group. Results:Forty-nine patients was diagnosed as DISH with the prevalence of 37.1% in all included cases. The prevalence was about twice as high in male (46.3%) than in female (23.1%) ( χ2=8.806, P=0.003). The prevalence in the age groups of <40, 40-49, 50-59, 60-69 and ≥70 years was 20.0%, 28.0%, 34.4%, 44.0%, and 66.7%, respectively. The prevalence in long-segment TOLF patients (45.1%) was significantly higher than that in short-segment TOLF patients (24.0%) ( χ2=5.937, P=0.015). DISH most frequently affected T 8,9 levels (91.8%). The total number and mean number of ossified segments were 365 and 7.4, respectively. Ossification lesions in the upper, middle, lower thoracic spine accounted for 26.03%, 40.54%, and 33.15%, respectively. Grade I, grade II, and grade III ossification accounted for 21.4%, 28.5% and 50.1%, respectively. The mean age of the DISH group was older than the non-DISH group ( t=2.024, P=0.045). The proportion of male patients in the DISH group was significantly higher than that in the non-DISH group ( χ2=8.806, P=0.003). The average height and weight in the DISH group were significantly greater than those in the non-DISH group ( t=2.564, P=0.012; t=2.191, P=0.030), whereas no significant differences in BMI and constituent ratio of concurrent diabetes, cardiac disease, hypertension between two groups were observed. Conclusion:The prevalence of DISH in patients with TOLF is 37.1%. Male, elderly and long-segment TOLF patients are associated with higher prevalence. DISH frequently occurs in the middle and lower thoracic spine, and T 8,9 is the common affected segment. Ossification lesions may develop with age. Demographic characteristics of DISH group differ, to some extent, from those of non-DISH group.

Thoracic ossification of ligament flavum (TOLF) is a pathological process of heterotopic bone formation from ligamentum flavum tissue, which is the most common cause of thoracic spinal stenosis and thoracic myelopathy. Imaging examination is the predominant measure for localization and qualitative diagnosis for TOLF. Various TOLF classification systems have been reported currently, based on its morphology, distribution, configuration of compressed spinal cord or its association with peripheral tissue structures. However, there has been no unified classification due to confusing clinical applications. Therefore, we conducted a review to summarize TOLF classification systems and to evaluate the reliability of these classification systems and the diagnostic accuracy of various imaging modalities. As demonstrated in literatures, the diagnostic accuracy of radiography and the reliability of related classification were low. CT-based Sato classification (lateral, extended, enlarged, fused, tuberous) and Kuh classification (unilateral, bilateral, bridging), MRI-based Kuh classification (round, beak) and Chen Zhongqiang classification (isolated, skipping, continuous), Muthukumar classification combined with dural ossification (Tram track sign, Comma sign) elaborated ossification characteristics from different angles and dimensions. These classification systems were clinically significant in pathogenesis implication, risk assessment, treatment guidance and prognosis judgement, which showed high diagnostic accuracy and reliability. Combining multi-dimension and multi-level parameters of CT and MRI will play an important role in the diagnosis, treatment and new classification research of TOLF.

Ossification of posterior longitudinal ligament (OPLL) and ossification of ligamentum flavum (OLF) are clinically common heterotopic ossification diseases, which are the main causes of cervical and thoracic spinal stenosis and spinal cord injury. In some cases, OPLL and OLF may involve multiple spinal sites, individually or concurrently, increasing the difficulty of clinical diagnosis and treatment. This review initially attempts to define this specific ossification phenomenon as Tandem ossification of intraspinal ligaments (TOIL). It refers to a kind of severe spinal ligament ossification diseases caused by multiple factors, that OPLL and OLF occur alone or in combination at two or more sites of the cervical, thoracic, or lumbar spine with five or more ossified segments, progressively compressing the spinal cord or nerves, thereby resulting in a series of complex clinical symptoms. The prevalence of TOIL remains unknown, but its clinical detection rate is relatively high, and the most common TOIL is cervical OPLL combined with thoracic OPLL or/and OLF. Moreover, occurrence and development of TOIL involve many factors and its definitepathogenesis is not clear. Damage of upper and lower motor neurons can occur simultaneously in TOIL patients so that their clinical symptoms and signs often interfere with each other. TOIL has various forms, complex imaging features, and no uniform diagnostic and localization diagnostic criteria, which may easily lead to missed diagnosis and misdiagnosis. It is recommended that TOIL is identified by the typical symptoms and signs as the basis, CT three-dimensional reconstruction and MRI signs as the reference, and combination of the possible risk factors and clinical experience. Currently, surgical decompression for responsible lesions is the only effective treatment for symptomatic TOIL. However, domestic and foreign scholars still have great arguments on the optimal surgical strategy of TOIL. No consensus has been reached on how to determine the operating segment and the extent of decompression, which surgical approach and technique to be adopted, and when to select staged or one-stage combined surgery. Therefore, this review summarizes and discusses the current status and progress of the clinical characteristics and surgical strategies of TOIL through searching the relevant literature, to provide a reference for clinicians to diagnose and treat TOIL.

Objective: To investigate the mechanism of lncRNA HCG18/miR-17-5p/HMGA2 axis regulating the proliferation and metastasis of non-small cell lung cancer (NSCLC) cells. Methods: Sixty-two pairs of NSCLC tissues and corresponding para-cancerous tissues collected at Central Hospital of Chengde City from June 2017 to June 2018 were used for this study; in addition, NSCLC cell lines (A549, NCI-H1299, H1650, NCI-H460) and human lung epithelial BEAS-B cells were also collected. mRNA expression levels of HCG18, miR-17-5p and high-mobility group AT-hook 2 (HMGA2) in NSCLC tissues and cell lines were measured by quantitative Real-time polymerase chain reaction (qPCR). Si-HCG18, miR-17-5p, miR-17-5p+HCG18 or pcDNA3.1-HMGA2 were transfected into A549 cells and NCI-H460 cells; CCK-8 assay was used to detect the proliferation of transfected cells, Transwell assay was used to detect the migration and invasion ability of cells, and Wb was used to analyze the expressions of HMGA2 and EMT associated proteins (E-cadherin, N-cadherin and vimentin). The target relationships between HCG18 and miR-17-5p, or between miR-17-5p and HMGA2 were confirmed by dual luciferase reporter gene assay. Mice A549 cell xenograft model with HCG18 knockdown was constructed, and the growth of transplanted tumor was observed. Results: lncRNA HCG18 was highly expressed in NSCLC tissues and cells (all P<0.01); HCG18 level was significantly increased in patients at late stage or with lymphnode metastasis; and high HCG18 level was correlated with poor prognosis and low survival rates of NSCLC patients (all P<0.01). Knockdown of HCG18 significantly inhibited NSCLC cell proliferation, migration and invasion (all P<0.01), up-regulated E-cadherin expression but suppressed N-cadherin and vimentin expression (all P<0.01), and the volume of xenograft was obviously decreased (P<0.05). Dual luciferase reporter gene assay confirmed the relationship between HCG18 and miR-17-5p as well as miR-17-5p and HMGA2. miR-17-5p transfection significantly inhibited NSCLC cell proliferation, migration and invasion (all P<0.01), and up-regulated E-cadherin expression, reversely suppressed N-cadherin and vimentin expression (all P<0.01); however, miR-17-5p + HCG18 transfection reversed the effect of miR-17-5p on NSCLCcells.Conclusion:HCG18promotes the proliferationandmigrationofNSCLCcellsthrough regulating miR-17-5p/HMGA2 axis.