OBJECTIVE To study the effects of transferrin-targeting peptide T7 （7pep） on intracellular transportation of polyethylene glycol-polycaprolactone （PEG-PCL） micelles in human cervical cancer HeLa cells. METHODS Using coumarin-6 （C6） as fluorescent indicator probe， both coumarin-6 （C6）-loaded PEG-PCL （PEG-PCL-C6） micelles and 7pep-modified PEG- PCL （7pep-PEG-PCL-C6） micelles were prepared by film-dispersion method. The particle size， polydispersity index and appearance morphology were compared between two types of micelles； the real-time uptake of two types of micelles by HeLa cells was compared， and the colocalization of two types of micelles with early endosomes （EE）， endocytic recycling compartments （ERC） and late endosomes （LE） after entry into the cells was observed. RESULTS The particle sizes of PEG-PCL-C6 and 7pep-PEG-PCL- C6 micelles were（75.0±2.3）and（82.0±1.5）nm； the polymer dispersity indexes were 0.17±0.20 and 0.17±0.32， respectively， with a regular spherical appearance. The colocalization results showed that entry speed and amount of 7pep-PEG-PCL-C6 micelles were significantly faster/more than those of PEG-PCL-C6 micelles. 7pep-PEG-PCL-C6 micelles entered EE faster than PEG-PCL-C6 micelles， while PEG-PCL-C6 micelles entered ERC at a faster rate than 7pep-PEG-PCL-C6 micelles， and both PEG-PCL-C6 micelles and 7pep-PEG-PCL-C6 micelles tended to accumulate gradually in LE； Pearson coefficient， signal overlap ratio， and colocalization ratio of 7pep-PEG-PCL-C6 micelles with LE were significantly lower 60 minutes after entering the cell than those 30 minutes after entering the cell （P＜0.05 or P＜0.01）. CONCLUSIONS Targeting 7pep modification can increase the entry speed and amount of PEG-PCL-C6 micelles， and also alter their intracellular transportation behavior.

This paper discussed how to set the limit of impurities in the quality standard when the impurities in small-molecular innovative drugs were metabolites.Firstly,through the analysis and interpretation of each relevant guideline,in combination with the review cases of FDA and EMA,found out the conditions for qualification of im-purity,and then establ ish the acceptable limit of impurities based on the test results of multiple batches,the incre-ments of production process and storage process.

Objective:To compare the efficacy of percutaneous vertebroplasty (PVP) under enhanced regional and conventional anesthesia for multisegmental acute symptomatic osteoporotic thoracolumbar fractures (m-ASOTLF).Methods:A retrospective cohort study was conducted to analyze the data of 91 patients with m-ASOTLF who were admitted to Honghui Hospital of Xi′an Jiaotong University from January 2021 to December 2022, including 36 males and 55 females, aged 55-80 years [(67.4±7.3)years]. According to American Society of Anesthesiologists (ASA) classification system, 18 patients were classified as grade I, 52 grade II, and 21 grade III. Injured segments included T 6-T 10 in 23 patients, T 11-L 2 in 47 and L 3-L 5 in 21. All the patients were treated with PVP, among whom 45 were given enhanced regional anesthesia (enhanced anesthesia group) and 46 regional conventional anesthesia (conventional anesthesia group). The following indicators were compared between the two groups: the operation time, intraoperative bleeding, intraoperative heart rate, intraoperative mean arterial pressure (MAP), number of intraoperative fluoroscopies, and total amount of bone cement injected; the visual analogue scale (VAS) and Oswestry dysfunction index (ODI) before surgery, at 1 day, 1 month after surgery and at the last follow-up; the mini-mental state examination (MMSE) before surgery, at 1, 6, and 12 hours after surgery; the anterior vertebrae height (AVH), middle vertebrae height (MVH), and vertebral kyphosis angle (VKA) before and at 1 day after surgery; the incidence of complications such as bone cement leakage. Results:All the patients were followed up for 12-20 months [(15.8±2.6)months]. There were no significant differences between the two groups in the operation time, intraoperative bleeding, intraoperative heart rate, intraoperative MAP, number of intraoperative fluoroscopies or total amount of bone cement injected ( P>0.05). No significant differences were found between the two groups in VAS or ODI before surgery and at the last follow-up ( P>0.05). The VAS scores in the enhanced anesthesia group were (2.5±0.4)points and (1.8±0.3)points at 1 day and 1 month postoperatively respectively, which were both lower than (3.5±0.4)points and (2.0±0.5)points in the conventional anesthesia group ( P<0.01). The ODI values in the enhanced anesthesia group were 39.8±3.3 and 26.5±5.0 at 1 day and 1 month postoperatively respectively, which were both lower than 43.8±7.5 and 30.3±6.4 in the conventional anesthesia group ( P<0.01). The VAS and ODI at all postoperative time points decreased in both groups compared with those before surgery, with significant differences among those at all postoperative time points ( P<0.05). There was no significant difference between the two groups in the MMSE scores before, at 1, 6, and 12 hours after surgery ( P>0.05). The MMSE scores at 1 and 6 hours postoperatively were lower than that preoperatively in both groups ( P<0.05), and it was increased at 6 hours compared with that at 1 hour postoperatively ( P<0.05). There was no significant difference between the MMSE scores at 12 hours postoperatively and preoperatively in both groups ( P>0.05). The differences between the two groups in AVH, MVH, or VKA preoperatively were not statistically significant ( P>0.05). The AVH and MVH at 1 day postoperatively in the enhanced anesthesia group were (22.4±4.2)mm and (22.7±3.7)mm respectively, which were both higher than those in the conventional anesthesia group [(19.3±3.7)mm and (20.1±6.3)mm] ( P<0.05 or 0.01); the VKA at 1 day postoperatively in the enhanced anesthesia group was (13.9±3.7)°, which was lower than that in the conventional anesthesia group (15.8±4.1)° ( P<0.05). The AVH, MVH, and VKA in both groups were all improved at 1 day postoperatively compared with those preoperatively ( P<0.05). The incidence of bone cement leakage in the enhanced anesthesia group was 6.7% (3/45), which was lower than 21.7% (10/46) in the conventional anesthesia group ( P<0.05). Conclusion:Compared with conventional regional anesthesia, PVP under enhanced regional anesthesia for m-ASOTLF has more advantages in early postoperative pain relief, improvement of spinal function, restoration of vertebral height and reduction of bone cement leakage.

Objective To investigate the regulatory mechanism of apurnic/apyrimidinic endonuclease 1(APE1)in the transformation of chronic intestinal inflammation to colitis-associated colorectal cancer(CAC).Methods C64S mutant(APE1C64S)mice and APE1 wild type(APE1WT)mice were randomly divided into experimental group and control group.In vivo CAC model was established by azoxymethane(AOM)and dextran sulfate sodium salt(DSS)solution.Immunohistochemistry(IHC)and multiple IHC(mIHC)assays were used to observe the expression of APE1 and immune cell infiltration in colon tissues of each group.A mouse colon cancer cell line MC38 with stable knockdown of APE1 was constructed by lentivirus transfection,and subcutaneous tumor bearing experiments were performed in APE1WT and APE1C64S mice to confirm that tumor cell-derived APE1 caused immunosuppressive tumor microenvironment.The expression of APE1 and CXCL1[chemokine(C-X-C motif)ligand 1]and the infiltration of immune cells in tumor-bearing specimens were analyzed by IHC and mIHC assays.The tumor specimens of a 28-year-old female patient with CAC from Army Medical Center of PLA were analyzed for the expression of APE1 and CXCL1 and the infiltration of immune cells in the tumor and adjacent inflammatory tissues by IHC and mIHC assays.Results Compared with the control group and APE1WT erperimental group,APE1C64S erperimental group had significantly reduced disease activity index and tumor formation,polymorphonuclear myeloid-derived suppressor cells(PMN-MDSCs)infiltration,and CD4+and CD8+T cells(P＜0.05).No significant differences were observed in tumor growth and immune cells between APE1WT and APE 1C64S mice bearing subcutaneous tumors.However,in the tumor-bearing experiment using tumor cells with knockdown of APE1,the tumor growth was significantly lower and the number of infiltrated PMN-MDSCs was reduced,while those of CD4+and CD8+T cells were significantly increased(P＜0.05).Furthermore,high expression of APE1 and increased infiltration of PMN-MDSCs were found in the tumor tissues of the young CAC patient,and CD8+T cells were significantly reduced in the tumor tissues compared with the inflammatory tissues(P＜0.05).Conclusion APE1-redox in tumor cells can promote the infiltration of PMN-MDSCs and reduce the number of T cells,thereby forming an immunosuppressive tumor microenvironment and mediating the occurrence and development of CAC.

Objective:To investigate the positive rate and isolate Bordetella pertussis in children with suspected whooping cough and their close family members in Zhejiang Province, and further explore the susceptibility and resistant mechanism of Bordetella pertussis to antibiotics. Methods:A total of 273 nasopharynx swabs specimens from children with suspected whooping cough in Hangzhou Children′s Hospital from May 2022 to October 2022 were collected. The strains were isolated and cultured using charcoal select agar plate. Pertussis target genes were detected by RT-PCR. E-test method was used to detect the sensitivity of Bordetella pertussis strains to different antibiotics. The mechanism of resistance of Bordetella pertussis to macrolides was analyzed by whole genome sequencing. The phylogenetic analysis of isolated strains was based on core genome multilocus sequence typing(cgMLST). Results:Among 273 clinical samples of children with suspected pertussis and their close family members, 168 samples were positive by fluorescence quantitative PCR, accounting for 61.54%, and 30 pertussis strains were successfully isolated with a positive rate of 10.98%. In addition, among the 143 samples of close family members, 54.55% (78/143) samples were positive by RT-PCR and 9.79% (14/143) samples were positive by culture, suggesting that the close family member are important in family transmission of pertussis. Besides, most of the positive samples were from mothers. The results of E-test showed that 96.67%(29/30) strains showed high resistance to azithromycin with MIC value>256 mg/L, and the resistant mechanism of azithromycin was A2047G mutation in 23S rRNA. The phylogenetic analysis based on the cgMSLT showed that the isolated strains were clustered into two new different clades.Conclusions:The positive rate of Bordetella pertussis in close family members is at a high level and the mother may be the main source of infection, which is of great significance for monitoring and laboratory detection of suspected children′s family members. Bordetella pertussis shows high resistance to macrolides in Zhejiang Province, so monitoring of the antimicrobial resistance should be further strengthened to provide theoretical basis for clinical treatment and drug guidance.

Objective:To identify the phosphodiesterase (PDE) activity of the gene product encoded by LA_RS06960 of Leptospira interrogans ( L. interrogans), and analyze whether dinucleotides that can be degraded by PDE can activate macrophages to express innate immune factors. Methods:The LA_RS06960 gene in L. interrogans strain 56601 was amplified by PCR, and the prokaryotic expression system was constructed for the protein expression. The expressed rPDE was purified by Ni-NTA affinity chromatography. High performance liquid chromatography (HPLC) was used to measure the degration of c-di-AMP or 5′-pApA to AMP by rPDE. Real-time fluorescent quantitative RT-PCR (qRT-PCR) was used to detect the changes in the expression of target genes in Leptospira or THP-1 cells associated with innate immune factors during infection. qRT-PCR and ELISA were used to detect the changes in the expression and secretion level of the innate immune factors in macrophages treated with bacterial dinucleotide. Results:The prokaryotic expression system for LA_RS06960 gene of L. interrogans was constracted successfully, and the purified rPDE could degrate 5′-pApA and c-di-AMP into AMP in vitro. The mRNA level of leptospiral LA_RS06960 gene was significantly down-regulated, while IFN-β, TNF-α, IL-6 and IL-1β encoding genes of macrophages were significantly up-regulated during infection. The mRNA level or the secretion level of IFN-β and IL-6 of macrophages were increased after treated with the bacterial dinucleotide substrate of PDE. Conclusions:PDE encoded by LA_RS06960 gene has phosphodiesterase activity, and the bacterial dinucleotide substrate of the PDE could activate the innate immune response of macrophages.

Objective To analyze the efficacy and safety of double-ring sandwich procedure in the treatment of acute A-type aortic dissection root.Methods The clinical data of 103 patients with acute A-type aortic dissection treated by double-ring sandwich procedure in our center from October 2020 to May 2023 were retrospectively analyzed to investigate the surgical effect,short-term cardiac structural changes and short-term postoperative complications.Results Needle hemostasis was applied in 11.65％of all patients.The amount of red blood cell transfusion during the perioperative period was(2.95±1.97)U,plasma was(543.68±208.48)ml,and blood platelet was(0.08±0.30)U.The residual aortic regurgitation,cardiac function and pericardial effusion were significantly improved at 2 weeks after operation(P＜0.05).The proportion of residual false lumen in the aortic root was significantly reduced after operation(P＜0.05).The length of hospital stay,duration of intubation,and drainage within 24 hours after surgery were less.The incidence of postoperative short-term complications such as secondary thoracotomy,cerebral infarction,cerebral hemorrhage,hemofiltration,tracheal incision and death was low.Conclusions Double-ring sandwich procedure in the treatment of acute A-type aortic dissection can effectively block the false lumen of aortic root dissection,reduce the risk of aortic intimal avulsion,and the operation is safe and effective.

Objective To explore the relationship between the expression of lncRNA MGP-AS2 in bladder cancer tissues and the survival of bladder cancer patients,and to study the impact of MGP-AS2 on the biological function of bladder cancer cells and possible mechanism.Methods The TANRIC database was used to analyze MGP-AS2 expression level in bladder cancer tissues and its relation-ship with patient survival.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of MGP-AS2 in bladder cancer cell lines.The MGP-AS2 overexpression plasmid and negative control(NC)plasmid were transfected into bladder cancer 5637 cells,respectively,as the MGP-AS2group and NC group.Plate cloning experiment,cell scratch experiment and Tr-answell experiment were used to analyze the effects of MGP-AS2 overexpression on the proliferation,migration and invasion of bladder cancer 5637 cells.Dual-luciferase reporter gene experiment was used to verify the targeted binding between MGP-AS2 and miR-18a.The TANRIC database was used to analyze the correlation between MGP-AS2 and miR-18a expression.RT-qPCR method was used to analyze the effect of MGP-AS2 overexpression on the expression of miR-18a in bladder cancer 5637 cells.Western blot method was used to detect the effect of MGP-AS2 overexpression on the expression of Wnt/β-catenin signaling pathway proteins in bladder cancer 5637 cells.Results The expression level of MGP-AS2 in bladder cancer tissue was significantly lower than that in normal bladder tis-sues(P＜0.01).The overall survival and disease-free survival of patients with high expression of MGP-AS2 were significantly longer than those of patients with low expression of MGP-AS2(P＜0.01).The expression level of MGP-AS2 in bladder cancer cell lines were lower than those in SV-HUC-1 cells(P＜0.05).The bladder cancer cells with the lowest MGP-AS2 expression was 5637 cells(P＜0.01).Compared with the NC group,overexpression of MGP-AS2 could significantly inhibit the proliferation,migration and invasion a-bility of 5637 cells(P＜0.01).Dual-luciferase reporter gene experiment confirmed that MGP-AS2 could target and bind to miR-18a(P＜0.01).Compared with the NC group,overexpression of MGP-AS2 could downregulate the expression of miR-18a(P＜0.01).After overexpression of MGP-AS2,Wnt/β-catenin signaling pathway proteins were all down-regulated in bladder cancer 5637 cells(P＜0.01).Conclusion MGP-AS2 is low-expressed in bladder cancer,and the expression level of MGP-AS2 is correlated with the prognosis of bladder cancer patients.MGP-AS2 may inhibit the transduction of the Wnt/β-catenin signaling pathway by adsorbing miR-18a,thereby inhibiting the progression of bladder cancer.

The discussion and research on the clinical dominant diseases of traditional Chinese medicine （TCM） have attracted increasing attention. Through approaches including modern technology， evidence-based medical methods， and multi-disciplinary treatment， we should construct a sound TCM inheritance and innovation system， establish a collaborative innovation mechanism， and integrate major research projects， striving to make breakthroughs in TCM theory， methodology， standards， and regulation system， promoting the scientific and technological progress of TCM， and thereby improving its curative effect. The China Association of Chinese Medicine （CACM） carried out a series of youth salon seminars for clinical dominant diseases in TCM， discussing and sorting out the advantages of the dominant diseases in clinical diagnosis and treatment of TCM and integrated traditional Chinese and western medicine in specific diseases or fields. Authoritative experts in the industry were invited to give comment and guidance to form a report. Centering on clinical research of dominant diseases， thematic research was carried out in the aspects of practice， human experience-based evidence， and transformation path. Through the systematic study of the dominant diseases， the advantages of TCM in different stages of disease treatment were excavated to constantly improve the prevention and treatment ability of TCM and carry forward the advancement of TCM theory and practice. At the same time， the communication and understanding between traditional Chinese and western medicine were improved， laying the foundation for the further formation of industry guidelines or consensus and comprehensive promotion. These seminars are expected to provide references for the development of policy planning， clinical diagnosis and treatment， health economy， and social services in TCM and lay the foundation for the formation of a new modern diagnosis and treatment system with Chinese characteristics.

Objective:To compare the clinical efficacies of robot-assisted and free-hand long segment screw fixation combined with wedge osteotomy in the treatment of type IV chronic symptomatic osteoporotic thoracolumbar fractures (CSOVCFs).Methods:A retrospective cohort study was conducted to analyze the clinical data of 72 patients with type IV CSOVCFs who were admitted to Honghui Hospital of Xi′an Jiaotong University from May 2019 to December 2021, including 22 males and 46 females; aged 61-82 years [(71.2±12.3)years]. Fracture segments were located at T 11-T 12 in 37 patients and at L 1-L 2 in 31. A total of 32 patients were treated with robot-assisted long segment screw fixation combined with wedge osteotomy (robot group) and 36 with free-hand long segment screw fixation combined with wedge osteotomy (free-hand group). The operation time, intraoperative bleeding volume, dosage of radiation exposure, intraoperative needle adjustment, time of single pedicle screw placement and accuracy of pedicle screw placement were compared between the two groups. The kyphotic Cobb angle, sagittal vertical axis (SVA), thoracic kyphosis (TK), lumbar kyphosis (LL), visual analogue scale (VAS) and Oswestry disability index (ODI) were measured preoperatively, at 3 days postoperatively and at the last follow-up. The incidences of facet joint violation, deviation in guide needle placement, cerebrospinal leak and proximal junctional kyphosis (PJK) were observed. Results:All patients were followed up for 12-26 months [(18.2±5.1)months]. The operation time and time of single pedicle screw placement showed no significant differences between the two groups (all P>0.05). The intraoperative bleeding volume was (502.5±58.3)ml in the robot group, less than that in the free-hand group [(690.2±45.9)ml]. The dosage of radiation exposure was (32.6±10.8)μSv in the robot group, lower than that in the free-hand group [(48.6±15.2)μSv]. The intraoperative needle adjustment was (2.1±0.3)times in the robot group, higher than that in the free-hand group [(20.7±5.8)times], and the accuracy of pedicle screw placement was 99.7% in the robot group, less than that in the free-hand group (91.8%) (all P<0.01). Compared with pre-operation, the kyphotic Cobb angle, SVA, TK and LL were significantly improved in both groups at postoperative 3 days and at the last follow-up (all P<0.05). Compared with postoperative 3 days, the kyphotic Cobb angle, SVA and TK were increased at the last follow-up within the two groups, but with no significant differences (all P>0.05). Compared with postoperative 3 days, the LL was decreased within the two groups at the last follow-up, but with no significant differences (all P>0.05). The VAS and ODI in the two groups were significantly lower at postoperative 3 days and at the last follow-up when compared with those before operation (all P<0.05), and both values were significantly lower at the last follow-up than those at postoperative 3 days (all P<0.05). There were no significant differences in the VAS or ODI at all time points between the two groups (all P>0.05). The incidence of facet joint violation in the robot group was 1.6%, markedly lower than that in the free-hand group (9.6%) ( P<0.01). The incidences of deviation in guide needle placement, cerebrospinal leak and PJK showed no differences between the two groups (all P>0.05). Conclusion:For type IV CSOVCFs, the robot-assisted long segment screw fixation combined with wedge osteotomy can better reduce intraoperative blood loss, decrease radiation exposure, improve accuracy of pedicle screw placement, and reduce facet joint violation when compared with free-hand long segment screw fixation combined with wedge osteotomy.