Objective To analyze the composition characteristics and biological functions of tumor cell subpopulations in glioblastoma(GBM)through multi-transcriptomics sequencing technology,and explore the hypoxia characteristics and spatial localization features of the mesenchymal-like(MES-like)tumor cell subpopulation in GBM and the influence on malignant biological behaviors.Methods Multi-transcriptomics sequencing data,including single-cell RNA sequencing(scRNA-seq)data(18 patients),bulk RNA sequencing(bulk RNA-seq)and spatial transcriptomics(ST)data of GBM,were employed to define cell subpopulations in GBM,and Gene Ontology(GO)and Gene Set Enrichment Analysis(GSEA)were utilized to analyze their functions.The proportions and locations of cell subpopulations in bulk RNA-seq data were evaluated with BayesPrism deconvolution.Immunofluorescence assay was conducted for verification on 12 paraffin samples of GBM from patients who visited the neurosurgical department of our hospital from 2015 to 2023 and met the pathological diagnostic criteria for GBM(10 males and 2 females,at an average age of 53.50 years and a median age of 54.50 years).pySCENIC was applied to predict specific transcription factors of tumor cell subpopulations.Results Tumor cells in GBM were highly heterogeneous,and could be mainly divided into 4 subpopulations:astrocyte-like(AC-like),neural progenitor-like(NPC-like),oligodendrocyte progenitor-like(OPC-like)and MES-like.Differential gene analysis found that the MES-like tumor cells highly expressed vascular endothelial growth factor A(VEGFA),adrenomedullin(ADM),N-myc downstream regulated 1(NDRG1),insulin like growth factor binding protein 5(IGFBP5),and A-kinase anchoring protein 12(AKAP12)(P<0.001).pySCENIC transcription factor prediction found that the high-active transcription factors of the MES-like tumor cells were AT-rich interaction domain 3A(ARID3A),FOS like 2,AP-1 transcription factor subunit(FOSL2),endothelial PAS domain protein 1(EPAS1),CCAAT enhancer binding protein delta(CEBPD),and CCAAT enhancer binding protein beta(CEBPB)(P<0.05).GO and GSEA enrichment analyses found that the MES-like tumor cells were enriched in hypoxia-related pathways,especially the pathway of cell responses to hypoxia levels(NES=2.437,P<0.001).BayesPrism deconvolution showed that the MES-like tumor cells mainly existed in PAN(Pseudopalisading cells around necrosis)and perinecrotic zone.Immunofluorescence assay confirmed CD44+(CD44 antigen)MES-like tumor cells were mainly located in hypoxia areas with highly expression of hypoxia inducible factor 1 subunit alpha(HIF1α)(P<0.01).Multivariate Cox regression analysis indicated that the MES-like tumor cells were significantly correlated with the adverse prognosis of GBM patients(HR=1.71,95%CI:1.38～2.11,P<0.001).Conclusion Tumor cells in GBM are of highly heterogeneity.They could be mainly divided into 4 subpopulations:AC-like,NPC-like,OPC-like and MES-like.MES-like tumor cells,mainly locating in PAN and perinecrotic zone,are characterized by hypoxia,which can promote the malignant progression of GBM.

Objective To investigate the role of homologous genes absent from the wings of drosophila melanogaster(Notch)signaling pathway in the imbalance of helper T cells 1(Th1)and helper T cells 2(Th2)and the intervention mechanism of Qizhi Zhoufei Granule in chronic obstructive pulmonary disease(COPD).Methods Ten of seventy Wistar rats were selected as the blank control group,and the other rats were established by cigarette smoking combined(CS)with tracheal infusion of lipopolysaccharide(LPS).The COPD model was established by randomly selecting 3 rats in the control group and the model group to verify the success of the model.At the end of modeling,gavage administration was performed.The rats in the model group were randomly divided into model control group,positive control group(67.5 μg·kg-1)and Qizhi Zhoufei Granule high,medium and low treatment group(3.24,1.62,0.81 g·kg-1).Each group was treated with normal saline,dexamethasone acetate suspension and Qizhi Zhoufei Granule suspension at high,medium and low doses.The rats in the blank control group were given the same volume of normal saline as the model control group.After modeling with 28 days and treatment with 28 days,peak inspiratory flow(PIF)and peak expiratory flow(PEF)were detected by the animal lung function test system.Rats were killed to extract lungs,spleen,serum and bronchoalveolar lavage fluid(BALF),hematoxylin-eosin(HE)staining was used to evaluate the pathological changes of lung tissues.The level of tumor necrosis factor-α(TNF-α)in serum and BALF was determined by enzyme-linked immunosorbent assay(ELISA).Flow cytometry was used to detect Th1/Th2 cells in spleen.Immunohistochemistry(IHC)and western blot were used to detect Notch1,Hes1 and Hey1 protein levels in lung tissues.Real-time fluorescence quantitative polymerase chain reaction(Real-Time PCR)was used to detect Notch1,Hes1 and Hey1 gene expression levels in lung tissues.Result Compared with the blank control group,the lung function of the model control group was significantly decreased(P<0.05),inflammatory cell infiltration and bronchial structure destruction occurred in the lung tissue,TNF-α content in serum and BALF increased significantly(P<0.05),the percentage of spleen Th1 cells was significantly decreased(P<0.05),and the percentage of Th2 cells was significantly increased(P<0.05),the protein and mRNA expressions of Notch1,Hes1 and Hey1 in lung tissues were significantly increased(P<0.05),the differences were statistically significant;Compared with the model control group,the lung function of rats in each administration group was significantly increased(P<0.05),the pathological injury of lung tissue was alleviated,TNF-α content in serum and BALF decreased significantly(P<0.05),the percentage of spleen Th1 cells was significantly increased(P<0.05),the percentage of Th2 cells was significantly decreased(P<0.05),the lung tissue of Notch1,Hes1,Hey1 protein and mRNA expression were significantly decreased(P<0.05),the differences were statistically significant.Conclusion Qizhi Zhoufei Granule regulate Th1/Th2 balance by inhibiting Notch signaling pathway,thereby improving pulmonary function and pathological injury,and affecting immune function in COPD rats.

Objective To explore the risk factors for suicidal ideation among the relatives and friends of suicide victims through psychological support hotline.Methods Calls from the relatives and friends of suicide victims to the Beijing psychological support hotline between 2009 and 2023 were included.Their current suicidal ideation was assessed by asking whether they had suicidal thoughts in the past two weeks.Demographic information of the callers was collected and the severity of their depression was assessed using the depression diagnostic screening scale.Interviews were conducted to evaluate distress,sense of hope,history of suicide attempts,and suicide risk factors.Logistic regression analysis was used to explore the risk factors for suicidal ideation among callers.Results A total of 360 cases were included in this study.There were 50.3%(181 cases)of the callers reporting suicidal ideation within 2 weeks before the call.Years of education(OR=0.86,95%CI:0.79-0.94)and high levels of hope(OR=0.19,95%CI:0.11-0.32)were protective factors against suicidal ideation in callers.History of suicide attempts(OR=2.01,95%CI:1.20-3.36),high levels of distress(OR=3.28,95%CI:1.92-5.61),and high levels of depression(OR=1.73,95%CI:1.05-2.84)were independent risk factors for suicidal ideation in callers.Conclusions History of suicide attempts,high levels of distress and high levels of depression among the relatives and friends of suicide victims calling the psychological support hotline are risk factors for their own suicidal ideation.

Objective:To summarize the clinical and genetic features of children with intellectual developmental disorder with seizures and language delay (IDDSELD) due to 12q24.31 deletion and SETD1B locus variants. Methods:The clinical data of a child with 12q24.31 deletion diagnosed in the Department of Pediatric Neurology of the Third Affiliated Hospital of Zhengzhou University in September 2022 were retrospectively analyzed. Trio-whole exome sequencing (trio-WES) and copy number variations sequencing (CNV-seq) were used for genetic analysis. The relevant literatures were reviewed to summarize the clinical features of the disease.Results:The proband was a 7 years and 9 month old girl who had clinical features of global developmental delay, epilepsy, hyperactivity, hypertonia, gait disorder, special facial features (high eyebrow arch, big ears, upper lip protrusion), funnel chest, lumbar lordosis. Karyotypic analysis showed 46XX in the proband. CNV-seq showed 12q24.31 (chr12: 121895654-122449092) position had a deletion of about 553.44 kb which contained the SETD1B gene. Trio-WES showed deletion of all exons 1-16 of the SETD1B gene. CNV-seq results of her parents were normal: the SETD1B gene was wild-type. This type has not been reported in China. Four children with IDDSELD caused by 12q24.31 deletion (including the SETD1B gene) were retrieved (totally 5 cases including this case), with male to female ratio of 1∶4, all with de novo mutations, and all with mental retardation, cephalo-facial and skeletal malformations. Three cases had seizures, 2 cases still had developmental backwardness after treatment, and 1 case was seizure controlled. Forty-seven cases of IDDSELD due to point mutation in the SETD1B gene were retrieved: male to female ratio was 31∶16, missense mutations (38/47) were predominant, most were de novo mutations (36/47), and a few were inherited from their fathers/mothers (6/47) or of unknown origin (5/47), with clinical manifestations of speech delay (43/47), growth retardation (43/47), intellectual disability (37/41), behavioral problems (37/47), facial malformations (34/47), skeletal malformations (23/47), obesity (16/47), skin abnormalities (11/47), etc. Thirty-nine cases were combined with seizures, 23 of whom were under control after treatment, and 8 cases were recorded as still having developmental backwardness after treatment. Conclusions:IDDSELD patients are rare at home and abroad, with diverse clinical phenotypes and difficult diagnosis. Symptomatic treatment is the main approach. And the patients can leave behind seizures and varying degrees of developmental backwardness. Among them, patients with 12q24.31 deletion are relatively rare and have not been reported in China, and this type is more common in females, all of whom have de novo mutations, and genetic testing is helpful for the early diagnosis of IDDSELD.

Chronic obstructive pulmonary disease （COPD） is characterized by long treatment course and poor prognosis. The pathogenesis has not been fully elucidated but is mostly related to the non-specific inflammation of the airway and surrounding tissues. T helper 1 （Th1） and T helper 2 （Th2） are generated by CD4⁺ T cell differentiation， and are in a dynamic equilibrium when the body is in normal state. The balance between pro-inflammatory cytokines and anti-inflammatory cytokines regulated by Th1/Th2 is vital for maintaining the immune homeostasis in respiratory tract. Chronic inflammatory state changes the level of inflammatory cells in the body， and there is immune disorder in T lymphocytes in the onset stage of COPD. Th1 cells are predominantly expressed in the stable stage of COPD， while Th2 cells are predominantly expressed in the acute exacerbations of COPD （AECOPD）. Th1/Th2 immune imbalance aggravates the inflammatory reaction， and thus restoring the immune balance between them and inhibiting the inflammatory reaction are critical for the treatment of COPD. At present， there has been no satisfactory treatment plan for COPD. Chinese medicine has a long history of preventing and treating COPD， with remarkable curative effect and few adverse reactions. A large number of animal experiments and clinical studies on Chinese medicine intervention of Th1/Th2 immune balance in COPD have indicated that Th1/Th2 immune balance is an important potential target for treating COPD by Chinese medicine， which can correct chronic inflammatory state by regulating the immune disorder of the body. It has also been found that Th1/Th2 balance plays an important immunoregulatory role in inflammatory response， but little is known about its specific mechanism in the pathogenesis of COPD. On this basis， this paper summarized and analyzed the biological characteristics of Th1/Th2 and their mechanism in the pathogenesis of COPD， as well as the intervention effect of single Chinese medicine or its effective components and Chinese medicine compound on Th1/Th2 immune balance in COPD. It further explored the pathogenesis of COPD and the potential therapeutic targets of Chinese medicine in interfering with Th1/Th2 immune balance in COPD， providing reference for further study on prevention and treatment of COPD with Chinese medicine.

Objective:To explore the clinical manifestations and genetic characteristics of Wolf-Hirschhorn syndrome (WHS) to improve the ability of diagnosis and differential diagnosis of the disease.Methods:The clinical features and auxiliary examinations and treatment of a proband with WHS caused by microdeletion of 4p16.3 segment who admitted to the Third Affiliated Hospital of Zhengzhou University in December 2021 were recorded, and whole exome sequencing (WES) of the family was performed. The prognosis was followed up.Results:The female proband, 11 months old, presented with convulsions at the age of 8 months, with the characteristics of heat sensitivity and cluster seizures, and her identical twin sister had a similar medical history. Physical examination found malnutrition, retarded development, special face, prominent forehead, wide nasal bridge, small jaw, precordial murmur and grade 3/6 murmur in the whole period, hyperactivity of P2, and low limb muscle tone. The whole exon and copy number variation (CNV) test of the family revealed that the proband had a 1.99 Mb heterozygous deletion in the chromosome 4p16.3 segment, including WHSC1 (NSD2), WHSC2 (NEFLA) and other genes. Copy number variation sequencing (CNV-Seq) of the proband and her sister showed 1.97 and 1.92 Mb heterozygous deletion of chromosome 4p16.3, respectively. Genealogical analysis by quantitative polymerase chain reaction revealed that the CNV was de novo, and it was determined to be a pathogenic variant according to the American College of Medical Genetics and Genomics guidelines. The proband took sodium valproate orally, and her sister took oral sodium valproate, zonisamide, and levetiracetam successively, and at the same time they received family rehabilitation training. The age at the last follow-up was 1 year and 8 months. Neither of them had convulsions again in the past 3 months, but the developmental delay was obvious. Conclusion:WHS patients may present with growth retardation, epilepsy, Greek warrior helmet-like special face, and congenital heart disease, and may have microdeletions in the chromosome 4p16.3 segment.

OBJECTIVE To provide reference for scientifi c and standardized development of clinical comprehensive evaluation of drugs in China. METHODS Guided by the theory of total quality management （TQM），drawing lessons from the successful experience of the British and German conducting evaluation ，combining with plan-do-check-act cycle and other quality management methods and tools ，drug clinical comprehensive evaluation of total quality management system was constructed in accordance with the requirements for our country related policy and local practice. RESULTS & CONCLUSIONS To construct total quality management system of clinical comprehensive evaluation of drugs in China from 5 aspects of organization system ，management process，assessment system ，evaluation and supervision platform ，support and guarantee mechanism. The organization system included national ，provincial and medical institutions ；management process should focus on the key links in the 3 stages of theme selection，evaluation and implementation ，and result transformation and application ；assessment system ，evaluation and supervision platform，support and guarantee mechanism should be established together so as to further improve the scientificity ，rationality， practicality and standardization of total quality management of clinical comprehensive evaluation of drugs. The development of total quality management is an effective starting point to promote the continual improvement of the drug clinical comprehensive evaluation；relevant government departments and the implementation of evaluation of medical institutions should further set up quality management consciousness ，establish report quality feedback mechanism and the results co-constructing and sharing mechanism and strengthen professional personnel training and innovation synergy regulation mode to ensure that the authenticity and reliability of evaluation results.

Objective:To summarize and analyze the clinical characteristics of different treatment in children with esophageal foreign bodies.Methods:This study collected 246 children with esophageal foreign bodies in our hospital from January 2016 to January 2020, which was divided into endoscopic group and operative group.The general and clinical data of children treated with different treatment were collected and statistical analyzed.Results:There were 222 children in endoscopic group and 24 children in operative group, respectively.The rate of surgery was 9.75%.There were no significant differences in gender and location of esophageal foreign bodies.However, the average age of operative group was(2.92±2.67) years, which significantly younger than that in endoscopic group(4.12±3.37)years( P=0.049). The residence time in operative group(median 29.10 h)was remarkable longer than that in operative group(median 11.80 h)( P<0.001). The proportion of sharpness(50.00%) and corrosive(45.83%) foreign bodies in operative group were more than those in endoscopic group[16.22% and 8.11%( P<0.001)]. Moreover, the occurrence rate of major complication in operative group was 83.33%, which was dramatically higher than that in endoscopic group(0.90%)( P<0.001). Conclusion:The younger and longer residence time of esophageal foreign bodies in children contribute to the rate of operative treatment.Additionally, the sharpness and corrosive foreign bodies increase the risk of surgery and serious complications.

Objective:The application of computer-rapid prototyping-assisted simulated hollow screw in the treatment of femoral neck fractures in fresh cadavers verified the reliability and accuracy of computer-rapid prototyping-assisted simulated hollow screw in the treatment of femoral neck fractures.Methods:The glass ion was labeled as a target for the treatment of femoral neck fracture with a cannulated screw. Three Kirschner pins were placed at the top and bottom of the greater trochanter. CT scan of the processed specimens. The scan data was imported into Mimics software, and the upper end of the femur was reconstructed three-dimensionally and the target of the femoral neck fracture was fixed with cannulated screws. The position of the cannulated screw in the femoral neck was designed. In Pro/E software, draw a guide that fits the three positioning Kirschner pins and cannulated screw guide needles on the upper end of the femur, and use rapid prototyping technology to make the guide. The navigation template was mounted on the positioning pin through the cannulated screw guiding hole, and the hollow screw guiding needle was drilled, and the hollow screw was inserted for the treatment of the femoral neck fracture to complete the positioning and guiding. According to the coordinates of the hollow screw guide needle and the target center in the Mimics software three-dimensional reconstruction coordinate system, the spatial distance calculation formula is used to calculate the distance between the hollow screw guide needle and the target center.Results:The coordinate analysis of the end point of the cannulated screw guide needle and the target can be obtained that the average distance between the end point of the cannulated screw guide needle for fixed femoral neck fracture and the target center point was 1.92 mm.Conclusion:The computer-rapid molding-assisted cannulated screw fixation of the femoral neck fracture is reliable and accurate, which provides a new idea and method for clinical individualization and precise fixation of the femoral neck fracture.

Objective:To summarize our experience in prevention of COVID-19 infection in emergency and confined operations during the first 3 weeks after Spring Festival in 2020.Methods:From February 3rd to 23rd, 2020, 151 patients were admitted to Department of Orthopaedic Trauma, Beijing Jishuitan Hospital for emergency and confined operations. In this cohort, 125 patients were admitted to ordinary wards. They were 70 males and 55 females with an age of 51.1 years ± 14.9 years. Of them, 2 were subjected to emergency operation and 123 to confined operation. The mean time from injury to operation was 9.9 days ± 6.1 days. There were 26 cases in the senile wards, 7 males and 19 females with an age of 80.8 years ± 7.0 years all of whom underwent confined operations. The mean time from injury to operation was 8.4 days ± 6.3 days. The protocols for emergency diagnosis, admission, emergency and confined operations, postoperative rehabilitation and management of suspects with COVID-19 during the epidemic of COVID-19 were optimized according to Diagnosis and Treatment Protocols for Novel Coronavirus Pneumonia (Trial version 5), emergency responding pre-plans of our hospital, and our experience in Enhanced Recovery After Surgery (ERAS) as well.Results:The patients in the ordinary wards had a hospital stay of 6.8 days ± 4.6 days while those in the senile wards 5.1 days ± 2.0 days. Abnormal temperature (≥37.3 ℃) was observed perioperatively in 17 cases in the ordinary wards. It was absorption fever in all and appeared in 4 cases upon admission. Fever appeared in 11 patients in the senile wards and upon admission in 3 of them. One senile patient who had been diagnosed of normal pneumonia returned to normal temperature and remained stable conditions after antibiotic therapy. The other patients were free of complications related to COVID-19 during their hospital stay.Conclusion:The first-line medical staff working at emergency department, wards and surgical theaters must heighten their vigilance against COVID-19 infection and rigorously follow protocols for prevention of COVID-19 infection in their daily clinical practice.