ObjectiveTo compare the differences between the ultrafiltration method and the dextran gel filtration method during the purification of Tfn-modified PCL micelles by using purification efficiency and micelle purity as indicators. MethodsCoumarin-6 （C6） was used as a fluorescent probe and was loaded into HOOC-PEG-PCL to form PCL micelles by the film-dispersion method. Tfn was then conjugated to the surface of PCL micelles via an amidation reaction， resulting in two types of micelles： Tfn/PCL_H and Tfn/PCL_L. The pharmaceutical properties of the two types of micelles were characterized. The micelles were then purified through ultrafiltration method and dextran gel method respectively， and the efficiency of the two methods， along with the purity of the final micelles， was compared. The density of Tfn on the surface of PCL micelles was also calculated. ResultsThe hydrated diameter of PCL micelles was approximately 73 nm， and the C6 loading efficiency was around 0.046%. The size increased to 134 nm and 158 nm for Tfn/PCL_L and Tfn/PCL_H， respectively. The micelle population was monodisperse. The purification results showed that， for the ultrafiltration method， after two and one rounds of purification， the Tfn/C6 ratio stabilized at 23.6 and 3.4 for Tfn/PCL_H and Tfn/PCL_L， respectively. For the dextran gel filtration method， the Tfn/C6 ratio reached 23.7 for the Tfn/PCL_H group after two rounds of purification. However， for the Tfn/PCL_L group， the Tfn/C6 ratio increased during four rounds of dextran gel purification， and a significant difference （P = 0.042 4） was observed between the first and last filtrations. The density of Tfn in the final micelles were calculated. For the ultrafiltration method， the Tfn density of Tfn/PCL_H and Tfn/PCL_L were 94.9% and 13.8%， respectively. For the dextran gel filtration method， the density of the two micelles were 95.6% and 14.4%， respectively. For Tfn/PCL_L group， the density results revealing a statistically significant difference （P=0.000 2）. ConclusionThe purification efficiency of the two methods is comparable. However， the purity of the final micelles shows a significant difference， with the dextran gel filtration method resulting in higher purity， particularly for the Tfn/PCL_L micelles.

Objective To explore the characteristics of upper airway obstruction on multi spiral computed tomography(MSCT)in patients with obstructive sleep apnea hypopnea syndrome(OSAHS)and their correlation with the disease,as well as to identify key characteristic indicators.Methods Forty-three patients with OSAHS(OSAHS group)and 45 normal controls(control group)were selected.MSCT of the upper airway was performed to analyze the differences in various observation indicators between the two groups,including diameters at levels of upper airway obstruction,soft palate thickness,mandibular plane to hyoid distance(MP-H),oropharyngeal length,air-pharyngeal ratio,and fat infiltration in the tongue base.A multivariate logistic regression model was con-structed to identify the main risk factors,and the receiver operating characteristic(ROC)curve was used to assess the diagnostic per-formance of the model.Results The OSAHS group exhibited smaller diameters at levels of upper airway obstruction and air-pharyn-geal ratio compared to the control group[area:(0.75±0.38)cm2 vs(1.36±0.58)cm2,t=5.855,P＜0.001].Additionally,body mass index(BMI),degree of fat infiltration in the tongue base,soft palate thickness,and MP-H were significantly greater in the OSAHS group than those in the control group(soft palate thickness:t=-6.881,P＜0.001).Multivariate logistic regression analysis revealed that fat infiltration in the tongue base,soft palate thickness,MP-H,and the area of upper airway obstruction were the main risk factors for OSAHS.The area under the curve(AUC)was 0.940[95％confidence interval(CI)0.868-0.979,P＜0.001],indica-ting good diagnostic performance.Conclusion MSCT can effectively evaluate the upper airway obstruction characteristics in patients with OSAHS,with fat infiltration in the tongue base,soft palate thickness,MP-H,and the area of upper airway obstruction as the primary indicators for OSAHS,providing a reference for clinical practice.

Objective To explore the characteristics of upper airway obstruction on multi spiral computed tomography(MSCT)in patients with obstructive sleep apnea hypopnea syndrome(OSAHS)and their correlation with the disease,as well as to identify key characteristic indicators.Methods Forty-three patients with OSAHS(OSAHS group)and 45 normal controls(control group)were selected.MSCT of the upper airway was performed to analyze the differences in various observation indicators between the two groups,including diameters at levels of upper airway obstruction,soft palate thickness,mandibular plane to hyoid distance(MP-H),oropharyngeal length,air-pharyngeal ratio,and fat infiltration in the tongue base.A multivariate logistic regression model was con-structed to identify the main risk factors,and the receiver operating characteristic(ROC)curve was used to assess the diagnostic per-formance of the model.Results The OSAHS group exhibited smaller diameters at levels of upper airway obstruction and air-pharyn-geal ratio compared to the control group[area:(0.75±0.38)cm2 vs(1.36±0.58)cm2,t=5.855,P＜0.001].Additionally,body mass index(BMI),degree of fat infiltration in the tongue base,soft palate thickness,and MP-H were significantly greater in the OSAHS group than those in the control group(soft palate thickness:t=-6.881,P＜0.001).Multivariate logistic regression analysis revealed that fat infiltration in the tongue base,soft palate thickness,MP-H,and the area of upper airway obstruction were the main risk factors for OSAHS.The area under the curve(AUC)was 0.940[95％confidence interval(CI)0.868-0.979,P＜0.001],indica-ting good diagnostic performance.Conclusion MSCT can effectively evaluate the upper airway obstruction characteristics in patients with OSAHS,with fat infiltration in the tongue base,soft palate thickness,MP-H,and the area of upper airway obstruction as the primary indicators for OSAHS,providing a reference for clinical practice.

Objective To screen items of the Clinical Aided Decision Scheme for Stroke Simultaneous Treatment of Disease,Pulse and Syndrome;To provide reference for the formulation and improvement of the scheme.Methods The Delphi method was used to distribute two rounds of questionnaires to 60 experts in cerebropathy or neurology across the country.Statistical analysis was performed on the questionnaire results of the scheme's items,including the disease names,etiology and pathogenesis,syndrome characteristics,rules and regulations,representative prescriptions,acupuncture and other therapies,and preventive care.Results Totally 42 and 50 valid questionnaires were collected.The experts reached the consensus for the importance of etiology and pathogenesis,rules and regulations,acupuncture and other therapies,and preventive care.In the section on syndrome characteristics,items with low relevance or causing ambiguity were removed.Items that were no longer used in modern times and different prescriptions with the same name were removed from the representative prescriptions.The names of syndromes,rules and regulations were unified.Conclusion The experts generally reached the consensus for the importance of the Clinical Aided Decision Scheme for Stroke Simultaneous Treatment of Disease,Pulse and Syndrome.However,there are still some limitations that require further study and discussion.

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.

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.

Primary central nervous system lymphoma (PCNSL) is a highly aggressive malignant lymphoma. As most chemotherapy drugs have difficulty in crossing the blood-brain barrier, PCNSL shows a difficulty in clinical treatment, a high recurrence rate and a poor prognosis. Early identification of relapsed patients and prompt initiation of salvage therapy play a critical role in the improvement of patients' prognosis. Brain biopsy is the gold standard to identify recurrence, while the risk of operation failure and complications is still high. Non-invasive imaging techniques are beneficial for early identification of recurrence in PCNSL and can provide an important basis for guiding relapsed patients to adjust treatment plans in time. However, there is no unified evaluation standard for imaging methods of monitoring the relapsed lesions of PCNSL. With the further research of the pathophysiological mechanism of PCNSL, biomarker detection has become a new method to identify recurrence and more clinical evidence is still needed in the future.

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 analyze the identification of pathogenic bacteria in municipal sewage of Hangzhou City, Zhejiang Province from January to October 2023, to understand the serotype and drug resistance of Vibrio cholerae isolated from sewage, as well as to predict the virulence gene and drug resistance gene carrying situation. Methods From January to October 2023, urban sewage samples from various environments in Hangzhou City, Zhejiang Province, were continuously collected for bacterial isolation and culture. The serotypes of 27 Vibrio cholerae strains were identified by slide agglutination method, and the drug sensitivity test was conducted by micro broth dilution method. Whole genome sequence and bioinformatics software were used to predict drug resistance genes and virulence genes. Results The sewage identification results showed 26 bacteria genera, including Salmonella, Vibrio, Enterobacter, and Aeromonas, among 82 bacterial species. All 27 strains of Vibrio cholerae were non-O1 and non-O139 serogroups. The strains showed high resistance rates to ampicillin, ampicillin/sulbactam and cotrimoxazole at 59.26%, 14.81%, and 14.81%, respectively), exhibited high sensitivity to cefotaxime, tetracycline, chloramphenicol, meropenem, ertapenem, ceftazidime, ceftazidime/avibactam, amikacin, ciprofloxacin, tigecycline, and azithromycin. A total of 21 different drug resistance genes were detected in 11 strains of Vibrio cholerae, among which the highest carriage rate of β-lactam resistance genes reached 63.64%, and 1 strain carried 17 kinds of drug resistance genes. None of the 11 strains of Vibrio cholerae carried ctxA, ctxB, ace, zot, or chxA genes, but all the strains carried rtxB, rtxC, and rtxD genes; 90.91% of the strains carried hlyA, and 36.36% carried rtxA. Conclusions All Vibrio cholerae isolated from sewage in this study are non-O1 and non-O139 serogroups, showing good sensitivity to most antibiotics, but also they carry a certain number of drug-resistance genes and virulence genes. This suggests that future monitoring efforts should focus on these strains, providing certain data support for the study of the characteristics of Vibrio cholerae isolated from sewage.

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.