Objective To establish an analytical method combining gas chromatography-tandem mass spectrometry(GC-MS)and liquid chromatography-tandem mass spectrometry(LC-MS)to detect diazepam residue in bait nest materials and fish samples,and improve the pretreatment steps of samples to make the experimental results accurate and the sample processing convenient and fast.Methods Taking diazepam as the research object,samples were extracted with methanol and dichloromethane/n-hexane as solvents according to the type,and the supernatant was taken for detection after centrifugation.Results The diazepam standard sample showed a good linear relationship in the range of 10～10 000 ng/mL.The retention times in methanol and mixed solvent were 13.54 min and 13.83 min,respectively,and the correlation coefficients were 0.998 and 0.999,respectively;The limit of detection(LOD)of using methanol as extraction solvent was 2 ppb,and limit of quantification(LOQ)was 6 ppb.The LOD of using mixed solvents was 5 ppb,and the LOQ was 15 ppb.When the sample is a bait nest material,the GC-MS spectrum was clear and standard,and the peak shape was sharp and prominent;When the sample is biological specimen,the GC-MS spectra are disturbed by matrix,while the LC-MS data is more accurate and faster.Conclusion It is more appropriate to use GC-MS to determine the content of bait nest materialsamples,and it is more accurate to LC-MS to determine the content of fish samples due to the complexity of the organism.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To observe the role of Huaiqihuang Granules (HQ) in the long-term management of bronchial asthma in young children, and the effective effect on concomitant rhinitis.Methods:A prospective real-world multicenter study was conducted in children aged 2-5 years with asthma diagnosed in the outpatient department (from April 2016 to March 2019)who received either inhaled corticosteroid (ICS)/leukotriene receptor antagonist (LTRA)(control group); inhaled ICS/LTRA plus HQ(combination group), or HQ alone(HQ group). All patients were followed up at week 4, 8, 12 after treatment. The number of days with asthma symptoms, the frequency of severe asthma attacks, the level of asthma control, and the days with rhinitis symptoms in the last 4 weeks were recorded. Differences before and after treatment, and those among groups after treatment were compared using Kruskal- Wallis H test or Wilcoxon rank-sum test. Results:A total of 2 234 eligible patients were recruited, and 2 147 cases completed followed-up visits, including 477, 1 374 and 296 cases in the control group, combination group, and HQ group, respectively. After the treatment, all 3 groups showed significant declines in the days with asthma symptoms, frequency of severe asthma attack and the days with rhinitis symptoms (all P<0.01), and the rate of well-controlled asthma increased significantly ( P<0.01). It lasted until the end of follow-up. Among groups, patients in the combination group showed significantly less days of asthma symptoms than those of the other 2 group at week 8 and 12[0(0, 0.9) d vs.0(0, 0.3) d, P<0.05; 0(0, 0.1) d vs. 0(0, 1.0) d, P<0.01]. Patients in the combination group and HQ group showed a significantly lower rate of severe asthma attacks than that of the control group at week 12 [0(0, 1), 0(0, 1), 0(0, 2), all P<0.05]. The well-controlled rate of asthma in the combination group was significantly higher than that of the control group and HQ group at week 8 and 12 (89.6% vs. 85.9% vs.82.1%, H=15.28; 90.9% vs. 84.1% vs. 81.8%, χ2=29.32, all P<0.01). Conclusions:HQ can significantly alleviate symptoms of asthma and rhinitis, severe attack of asthma, and increase the control rate of asthma when used as an additional treatment or used alone.

Mycobacterium neoaurum has the ability to produce steroidal intermediates known as 22-hydroxy-23, 24-bisnorchol-4-en-3-one (BA) upon the knockout of the genes for either the hydroxyacyl-CoA dehydrogenase (Hsd4A) or acyl-CoA thiolase (FadA5). In a previous study, we discovered a novel metabolite in the fermentation products when the fadA5 gene was deleted. This research aims to elucidate the metabolic pathway of this metabolite through structural identification, homologous sequence analysis of the fadA5 gene, phylogenetic tree analysis of M. neoaurum HGMS2, and gene knockout. Our findings revealed that the metabolite is a C23 metabolic intermediate, named 24-norchol-4-ene-3, 22-dione (designated as 3-OPD). It is formed when a thioesterase (TE) catalyzes the formation of a β-ketonic acid by removing CoA from the side chain of 3, 22-dioxo-25, 26-bisnorchol-4-ene-24-oyl CoA (22-O-BNC-CoA), followed by spontaneously undergoing decarboxylation. These results have the potential to contribute to the development of novel steroid intermediates.
Mycobacterium/metabolism* ; Phylogeny ; Steroids/metabolism* ; Metabolic Networks and Pathways ; Sterols/metabolism*

Objective: To establish the norm of the Physical Activity afterschool Questionnaire for Preschooler(P-PAQ) in urban areas of China, so as to provide a basis for graded guidance from the family perspective and to improve children s physical activity levels. Methods: From October 2020 to January 2021, 6 267 children aged 3-6 years old were recruited from 40 kindergartens in eight cities across six major administrative regions by stratified cluster sampling, and the P-PAQ initially developed by the researchers of this study were completed by the primary caregivers. The questionnaire was administered to collect data relating to the amount of physical activity undertaken by the preschoolers, and the norm was determined by quartiles. Data relating to parental concepts of sports and parental behavior were assessed by calculating mean scores in order to establish the norm. Results: Among preschoolers in urban areas, the M(P 25 ，P 75 ) of total physical activity time (min/day), moderate-to-vigorous physical activity time (min/day), outdoor time (min/day) and screen time (min/day) on school days outside kindergarten and on weekends were 84 (54,120), 22 (8,40), 12 (0,24) and 18 (6,30), and 170 (115,240), 60 (30,95), 90 (35,120) and 30 (20,60), respectively. When the score of parents sports concept and behavior (total score of 40) were≥34, 29-<34, 24-<29, <24, it was defined as four levels about above medium, medium, lower medium and lower, respectively. And for two dimensions，when the score of parental sports concept were ≥19, 17-<19, 15-<17, <15，and the score of parental behaviors were ≥16, 12-<16, 8-<12, <8, it was defined as four levels about upper medium, medium, lower medium and lower, respectively. Conclusion The norm of extracurricular activities among preschool children in Chinese cities has good representativeness and appropriate threshold values, which could provide a valuable reference for early assessment, as well as guidance in relation to out-of-school physical activity behaviors among children aged 3-6 years old.

Objective:To investigate the experience and perception of general practice residents on standardized training based on stratified progressive teaching model.Methods:From January to April 2022, focus group interviews were conducted among general practice residents who received standardized training at Peking University First Hospital.Results:Seventeen residents were selected by the purpose sampling method in the study, including 7 males and 10 females aged 24-39 years. Four themes were extracted from the interviews: lower levels of basic knowledge and feeling pressure at clinical work felt by junior residents; lack of clinical research ability and systematic training felt by senior residents; the rotation form and teaching contents were mono in grassroots practice bases; desire of raising salary levels for residents.Conclusions:In order to upgrade the quality of general practice residency training, it is necessary to strengthen the knowledge, skill training and research ability training of general practice residents, improve their treatment, and perfect the standardized training program accordingly.

Glycosite-specific antibody‒drug conjugatess (gsADCs), harnessing Asn297 N-glycan of IgG Fc as the conjugation site for drug payloads, usually require multi-step glycoengineering with two or more enzymes, which limits the substrate diversification and complicates the preparation process. Herein, we report a series of novel disaccharide-based substrates, which reprogram the IgG glycoengineering to one-step synthesis of gsADCs, catalyzed by an endo-N-acetylglucosaminidase (ENGase) of Endo-S2. IgG glycoengineering via ENGases usually has two steps: deglycosylation by wild-type (WT) ENGases and transglycosylation by mutated ENGases. But in the current method, we have found that disaccharide LacNAc oxazoline can be efficiently assembled onto IgG by WT Endo-S2 without hydrolysis of the product, which enables the one-step glycoengineering directly from native antibodies. Further studies on substrate specificity revealed that this approach has excellent tolerance on various modification of 6-Gal motif of LacNAc. Within 1 h, one-step synthesis of gsADC was achieved using the LacNAc-toxin substrates including structures free of bioorthogonal groups. These gsADCs demonstrated good homogeneity, buffer stability, in vitro and in vivo anti-tumor activity. This work presents a novel strategy using LacNAc-based substrates to reprogram the multi-step IgG glycoengineering to a one-step manner for highly efficient synthesis of gsADCs.

Promoting balanced distribution of medical resources and realizing high-quality sharing of basic medical services between urban and rural areas are an important part of common prosperity. Huzhou urban medical alliance was a new urban and rural of medical alliance mode based on level 4 vertical integration, which played an important role in the whole evolution process of the medical alliance. This medical alliance had not only broken through the " integration of counties" but also explored the " integration of cities" . It was a bridge connecting counties and cities, which was of great significance in narrowing the gap between urban and rural medical health services. The construction of urban medical alliance took digital transformation and systematic integration as two breakthroughs to comprehensively promote the upgrade of energy efficiency and benefit stacking of health governance. In 2021, the grass-roots medical utilization rate and county-level medical utilization rate were 72.7% and 90.6%. In 2020, the average hospitalization cost(8 726.7 yuan)and the average outpatient and emergency expenses(239.6 yuan)of public hospitals were 25.0% and 8.6% lower than the average level of Zhejiang Province, respectively. Although Huzhou city had broken through the restrictions of administrative divisions and actively promoted the construction of the medical alliance, and achieved phased results, there were still structural defects and institutional difficulties. In order to solve these problems, the author proposed to promote the improvement of governance structure and optimization of governance system through " three governance" and " three characteristics" , and then promote the high-quality development of urban medical alliances.

Objective : To investigate the effects of actin like 6A (ACTL6A) knockdown on the proliferation, apop⁃ tosis, migration and invasion of SW1990 cells in pancreatic cancer. Methods : The Oncomine database was used to analyze the expression of ACTL6A mRNA in the tissues of pancreatic cancer and normal pancreas. The plasmid of knockdown ACTL6A and siRNA negative control were established and transfected into SW1990 cell line as siRNA⁃ACTL6A group and siRNA⁃NC group. CCK⁃8, cell apoptosis experiment, Wound healing and Transwell assay were used to determine the effects of ACTL6A knockdown on the proliferation, apoptosis, migration and invasion of SW1990 cells. GSEA predicted a possible pathway regulated by ACTL6A in pancreatic cancer. T⁃test was used between the two groups. Results : The expression of ACTL6A in pancreatic cancer tissues was higher than that in normal pancreatic tissues ( P < 0. 05 ) . The results of CCK⁃8 assay showed that the absorbance of siRNA⁃ACTL6A group at 24 and 48 h were lower than those in the siRNA⁃NC group, and the difference was statistically significant ( t = 5. 840, 8. 454, P < 0. 01) . The results of Wound healing assay and Transwell assay showed that the healing rate and the number of invasive cells in siRNA⁃ACTL6A group were both lower than those in the siRNA⁃NC group. The difference was statistically significant ( t = 3. 960,4. 464, P < 0. 05), but the apoptosis rate of siRNA⁃ACTL6A group was significantly higher than that of the siRNA⁃NC group, and the difference was statistically significant( t = 12. 192, P < 0. 001) . GSEA results showed that the group with high expression of ACTL6A mRNA was up⁃regulated in cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway and other related gene sets(P < 0. 05) . These pathways were activated when the expression of ACTL6A was up⁃regulated. Conclusion ACTL6A is highly expressed in pancreatic cancer tissues. ACTL6A knockdown promotes the cell apoptosis of SW1990 cells, and inhibits proliferation, invasion and migration of SW1990 cells. The mechanism of the occurrence and development of ACTL6A in pancreatic cancer is attributed to the activation of cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway.

Objective:To detect the level of long non-coding RNA LINP1 (lncRNA-LINP1) in endometrial carcinoma and to explore the prognostic value of the expression leve.Methods:From January 2015 to December 2016, the tissue samples of 82 patients with endometrial carcinoma in North China University of Science and Technology Affiliated Hospital were used as endometrial carcinoma group, and the normal adjacent tissues were selected as the paracancerous control group.The expression of LINP1 mRNA was measured by real-time fluorescence quantitative PCR (qRT-PCR). The clinicopathological data of the patients were collected, and the relationship between the expression of LINP1 mRNA and clinicopathological parameters was analyzed.Kaplan-Meier method was used to analyze the relationship between the expression of LINP1 mRNA and the survival rate of endometrial patients; and COX multivariate analysis was used to identify the risk factors of death in patients with endometrial cancer.Results:The expression of LINP1 mRNA in endometrial carcinoma group and paracancerous control group were (2.38±0.43) and (1.00±0.24). There was significant difference between the two groups ( t=25.376, P<0.001). The results of clinicopathological parameters showed that the expression of LINP1 mRNA was not related to the age and histological type of endometrial carcinoma (all P>0.05). It was related to lymph node metastasis, FIGO stage, tumor differentiation and vascular invasion (all P<0.05). the three-year total survival rate of the group with high expression of LINP1 mRNA (48.78%) was lower than that of the group with low expression of LINP1 mRNA (80.49%), and the differences were statistically significant (χ 2=6.306, P<0.05). COX Regression analysis showed that, high expression of LINP1 mRNA and FIGO Stage III, IV and lymph node metastasis were the risk factors for death in endometrial cancer patients (HR(95% CI) were 2.898(2.031-4.136), 1.831(1.448-2.316), 1.708(1.364-2.138), P values were 0.002, 0.004 and 0.008, respectively. Conclusion:The high expression of LINP1 mRNA in endometrial carcinoma is closely related to lymph node metastasis, FIGO stage, tumor differentiation, vascular invasion and prognosis.It is a risk factor for the death of patients with endometrial cancer and may be used as an index to evaluate the prognosis of patients with endometrial cancer.