Objective: To investigate whether membrane vesicles (MVs) of Streptococcus mutans （S.mutans） contain autoinducer-2 (AI-2) and to preliminarily explore the effects of these MVs on the growth and biofilm formation of S. mutans. Methods: MVs were isolated from the S. mutans UA159 strain using differential centrifugation. The isolated MVs were characterized by nanoparticle tracking analysis for particle size and concentration and observed by transmission electron microscopy. The presence of AI-2 was identified using the Vibrio harveyi BB170 bioluminescence assay: the BB170 diluent was supplemented with AB medium (control group), MV extract (MVs group), pre-ultrafiltration supernatant (Sup group), or post-ultrafiltration supernatant (Sup-af group). The effects of MVs on growth and biofilm formation were assessed using the S.mutans UA159 strain or a luxS deletion mutant as the control group, compared with experimental groups stimulated with gradient concentrations of MVs (MVs-2.0E+7, MVs-2.0E+8, and MVs-2.0E+9 groups). Growth curves, MTT assay, and colony-forming unit (CFU) counts were used to determine changes in growth capacity. Biofilm formation was evaluated using crystal violet staining, confocal laser scanning microscopy, and the anthrone method for polysaccharide quantification. Results: Enriched S. mutans MVs were successfully obtained, with an average particle size of approximately 94.19 nm and a concentration of 1.87E+11 particles/mL. The bioluminescence assay showed that the luminescence intensity of the Sup group was higher than that of the Sup-af group, and the MVs group exhibited higher intensity than the control group. Assessments via growth curves, MTT assay, and CFU counts indicated no significant differences in the growth capacity of the various S. mutans strains after treatment with different concentrations of MVs. Crystal violet staining quantification and confocal laser scanning microscopy observations revealed that high-concentration MV treatment (2.0E+9 particles/mL group) resulted in lower biofilm mass compared to the control. The anthrone method showed that the production of both water-soluble and water-insoluble polysaccharides was significantly lower in the high-concentration MV group than in the control. Conclusion S. mutans MVs contain the quorum sensing signal molecule AI-2. These MVs do not significantly affect the growth of S. mutans, but they can regulate biofilm formation and exhibit an inhibitory effect at high concentrations.

[摘 要] 目的：探讨精氨酸-甘氨酸-天冬氨酸（RGD）修饰对肿瘤抑素19肽（T-19）抗肝癌活性的影响，比较分析T-19及RGD修饰的T-19（RGD-T-19）对肝癌SK-Hep-1细胞增殖、侵袭和迁移能力的影响。方法：用Fmoc固相法合成T-19及RGD-T-19，用高效液相色谱仪和质谱进行分离、鉴定。常规培养SK-Hep-1细胞，用0、50、100、150、200、250 mg/mL的T-19及RGD-T-19分别处理细胞，分为0 mg/mL（对照）组、50 mg/mL组、100 mg/mL组、150 mg/mL组、200 mg/mL组、250 mg/mL组。CCK-8法、克隆形成实验、划痕愈合实验和Tanswell小室实验、WB法和qPCR法分别检测SK-Hep-1细胞的增殖、迁移、侵袭能力，以及环氧合酶-2（COX-2）、基质金属蛋白酶-2（MMP-2）、MMP-9、组织基质金属蛋白酶抑制剂-1（TIMP-1）、TIMP-2蛋白和MMP-1、MMP-2 mRNA的表达。结果：经质谱鉴定，用Fmoc固相法合成的T-19及RGD-T-19纯度高。T-19和RGD-T-19均能显著抑制SK-Hep-1细胞的增殖、迁移、侵袭能力，抑制COX-2蛋白、MMP-2和MMP-9蛋白及mRNA的表达、促进TIMP-1、TIMP-2蛋白的表达（P < 0.05, P < 0.01, P < 0.001），RGD-T-19的抑制或促进效应均明显强于T-19（均P < 0.05）。结论：利用Fmoc固相法合成了纯度高、活性好的T-19及RGD-T-19，两种肽均能抑制SK-Hep-1细胞增殖、侵袭和迁移能力，RGD-T-19作用明显强于T-19。

The diagnosis and treatment resources for rare diseases in China are highly imbalanced. The basic diagnosis and treatment capabilities are weak, the diagnosis period for patients is long, and the rates of missed diagnosis and misdiagnosis are relatively high. The establishment of a hierarchical diagnosis and treatment system is the inevitable approach to enhancing the diagnosis and treatment standards of rare diseases. Currently, the implementation of the domestic hierarchical diagnosis and treatment system for rare diseases still confronts numerous challenges, such as ambiguous referral standards and processes of primary medical institutions, and ineffective information interaction among institutions at all levels. Thus, it is essential to facilitate high-level information construction for the hierarchical diagnosis and treatment of rare diseases. This paper explores the process of constructing a multidisciplinary joint remote diagnosis and treatment platform and a health management platform through informatization, with the hope of establishing two closed loops of digital diagnosis and treatment services and health follow-up management for patients with rare diseases, as well as achieving timely diagnosis and lifelong health management for patients. It integrates and optimizes auxiliary diagnostic tools, promotes the rapid dissemination of rare disease diagnosis and treatment experiences to the grassroots, enhances the information construction level of the hierarchical diagnosis and treatment system, and endeavors to address the practical predicament of weak diagnosis and treatment capabilities of rare diseases in grassroots medical institutions. Additionally, this paper proposes an essential approach for multi-dimensional independent innovation to guide the popularization of efficient and high-quality rare disease diagnosis and treatment services. By encompassing innovating the rare disease diagnosis and treatment collaboration network and multidisciplinary diagnosis and treatment model, facilitating the application of the latest biomedical and informatics technologies to the grassroots, and constructing a national intelligent data platform for rare disease innovation, a new model for rare disease services with Chinese characteristics will be established. This will significantly enhance the medical treatment level of rare diseases in China and strive for more benefits for patients.

Objective To investigate the effect of programmed death ligand-1(PD-L1)overexpression on epithelial mesenchymal transformation(EMT)in endometrial cancer cells and its possible mechanism.Methods Ishikawa/PD-L1,a PD-L1 stable overexpression cell line constructed in the laboratory and the control Ishikawa/EV were used.The efficiency of PD-L1 overexpression was detected by qPCR and Western blot assay.The cell migration ability and invasion activity were detected by scratch assay and Transwell assay.The EMT-related protein and the phosphorylation level of nuclear transcription factor-κB(NF-κB)were detected by Western blot.Results Compared with the control group,the migration ability and invasion activity of Ishikawa/PD-L1 were significantly enhanced,and the expression of E-cadherin was significantly down-regulated,whereas vimentin,N-cadherin and p-p65 were significantly increased.Conclusion PD-L1 can promote EMT by inducing the activation of NF-κB pathway,and thus enhance the migration and invasion ability of endometrial cancer cells.

Objective To analyze the efficacy and safety of camrelizumab combined with tegafur, gimeracil and oteracil potassium capsule in treating advanced esophageal squamous cell carcinoma. Methods Eighty patients with advanced esophageal squamous cell carcinoma treated in the hospital from July 2021 to June 2022 were divided into tegafur, gimeracil and oteracil potassium group (n=44) and combined group (n=36) according to the therapeutic methods. The tegafur, gimeracil and oteracil potassium group was orally treated with tegafur, gimeracil and oteracil potassium capsule, while the combined group was treated with camrelizumab injection on the basis of the tegafur, gimeracil and oteracil potassium group. Tumor markers and immune cell population levels were compared between the two groups before and after treatment; the quality of life was evaluated in both groups; the therapeutic effect and adverse reactions of drugs were analyzed between the two groups. Results After treatment, the squamous cell carcinoma antigen(SCC-Ag), cytokeratin 19 fragment (CYFRA21-1), carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125), CD8⁺, and the performance status of the Eastern Cooperative Oncology Group (ECOG) decreased significantly in both groups, while CD3⁺, CD4⁺, CD4⁺/CD8⁺, score of the Quality of Life (QOL), and score of the Karnofsky Scale increased significantly, and the improvement of the above indicators in the combined group were significantly greater than those in the tegafur, gimeracil and oteracil potassium group (P<0.05). The effective rate was 34.09% and the disease control rate was 72.73% in the tegafur, gimeracil and oteracil potassium group, while the effective rate was 63.89% and the disease control rate was 83.33% in the combined group; the effective rate of the combined group was significantly higher than that of thetegafur, gimeracil and oteracil potassium group (P<0.05). Both the tegafur, gimeracil and oteracil potassium group and the combined group occurred adverse reactions such as gastrointestinal discomfort, cardiovascular adverse reactions, myelosuppression, hair loss, thyroid dysfunction, rash and edema, but there were no significant differences between the two groups (P>0.05). The mean progression-free survival (PFS) was (3.13±0.94) months and the mean overall survival (OS) was (6.05±2.38) months in the tegafur, gimeracil and oteracil potassium group, while the mean PFS was (5.08±1.67) months and the mean OS was (9.01±3.54) months in the combined group; the mean PFS and OS in the combined group were significantly longer than those in the tegafur, gimeracil and oteracil potassium group (P<0.05). Conclusion Camrelizumab combined with tegafur, gimeracil and oteracil potassium capsule can effectively alleviate clinical symptoms, reduce pain and improve the quality of life in patients with advanced esophageal squamous cell carcinoma.

Objective: To examine the feasibility, safety, and efficacy of mobilization of the vertebral artery for C₂ pedicle screws in cases with high-riding vertebral artery (HRVA). Methods: The clinical data of 12 patients with basilar invagination and atlantoaxial dislocation underwent atlantoaxial reduction and fixation in the Department of Neurosurgery, the First Affiliated Hospital of University of Science and Technology of China between January 2020 and November 2021 were retrospectively analyzed. All patients had high-riding vertebral artery on at least one side that prohibited the insertion of C₂ pedicle screws. There were 2 males and 10 females aged (48.0±12.8) years (range: 17 to 67 years). After correction of vertical dislocation during the operation, the C₂ pedicle screw insertion and occipitocervical fixation and fusion were performed using the vertebral artery mobilization technique. Neurological function was assessed using the Japanese Orthopedic Association (JOA) scale. The preoperative and postoperative JOA score and the main radiological measurements, including the anterior atlantodental interval (ADI), the distance of the odontoid tip above the Chamberlain line, the clivus-canal angle, were collected and compared by paired t-test. Results: Mobilization of the high-riding vertebral artery was successfully completed, and C₂ pedicle screws were then fulfilled after the vertebral artery was protected. There was no injury to the vertebral artery during the operation. Meanwhile, no severe surgical complications such as cerebral infarction or aggravated neurological dysfunction occurred during the perioperative period. Satisfactory C₂ pedicle screw placement and reduction were achieved in all 12 patients. All patients achieved bone fusion 6 months after surgery. No looseness and shift in internal fixation or reduction loss was observed during the follow-up period. Compared to the preoperative, the postoperative ADI decreased from (6.1±1.9) mm to (2.0±1.2) mm (t=6.73, P<0.01), the distance of the odontoid tip above the Chamberlain line decreased from (10.4±2.5) mm to (5.5±2.3) mm (t=7.12, P<0.01), the clivus-canal angle increased from (123.4±11.1) ° to (134.7±9.6) ° (t=2.50, P=0.032), the JOA score increased from 13.3±2.1 to 15.6±1.2 (t=6.99, P<0.01). Conclusion: The C₂ pedicle screw insertion assisted by mobilization of the vertebral artery is safe and considerably effective, providing a choice for internal fixation in cases with high-riding vertebral arteries.

Objective To investigate the influencing factors of willingness to screen for cervical cancer in women in Chizhou. Methods A cross-sectional web-based survey of 3,058 migrant women living in Chizhou was conducted through a social media platform. The survey included questionnaires on socioeconomic characteristics, history of cervical cancer screening, knowledge of cervical cancer prevention and treatment, and attitude. The χ² test and logistic regression were used to analyze the influencing factors of willingness to screen. Results Under 60 years of age, having a spouse or sexual partner (OR=1.368), having a junior high school education or above, living in urban areas (OR = 1.501), local household registration (OR=1.363), high monthly income, and high knowledge of cervical cancer prevention and control (OR=1.198) were positively associated with the rate of willingness to undergo cervical screening. Among attitude barriers, there were statistically significant differences in fear of seeing male doctors (38.23%), fear of seeing poor results (31.92%), belief that screening was painful (23.56%), shyness or embarrassment (20.18%), and belief that they did not need to participate in screening due to asymptomatic (17.56%), having received HPV vaccine (15.86%), or ages at which screening was not considered necessary (15.14%) between the groups (P<0.05). Conclusion Socioeconomic characteristics, knowledge of prevention and treatment, fear of screening experience, and lack of motivation are related factors affecting cervical cancer screening willingness. Achieving cervical cancer elimination requires action to improve screening coverage in socio-economic, health education and women's attitudes.

Abstract By reviewing the advocacy framework and implementation path of global adolescent health promotion, summarizing the main health problems of adolescents worldwide and current status of adolescent health care services in China, an ecological model of adolescent health care was concluded according to the demands of adolescents towards health care services. A comprehensive intervention strategy, ie. multi sector cooperation, community participation and integration of hospital, school, community and family, etc, has been put forward to promote the health and sustainable development of adolescents.

RNA viruses are critically dependent upon virally encoded proteases to cleave the viral polyproteins into functional proteins. Many of these proteases exhibit a similar fold and contain an essential catalytic cysteine, offering the opportunity to inhibit these enzymes with electrophilic small molecules. Here we describe the successful application of quantitative irreversible tethering (qIT) to identify acrylamide fragments that target the active site cysteine of the 3C protease (3C^pro) of Enterovirus 71, the causative agent of hand, foot and mouth disease in humans, altering the substrate binding region. Further, we re-purpose these hits towards the main protease (M^pro) of SARS-CoV-2 which shares the 3C-like fold and a similar active site. The hit fragments covalently link to the catalytic cysteine of M^pro to inhibit its activity. We demonstrate that targeting the active site cysteine of M^pro can have profound allosteric effects, distorting secondary structures to disrupt the active dimeric unit.

AIM: To investigate the impact of intravitreal conbercept injection on the aqueous humor levels of vascular endothelial growth factor(VEGF), interleukin-6(IL-6)and interleukin-8(IL-8)in patients diagnosed with neovascular glaucoma(NVG), and to evaluate the efficacy of conbercept in combination with different surgical modalities.

METHODS: This study was conducted as a retrospective, case series investigation. A total of 102 patients(102 eyes)diagnosed with NVG from Jan. 2019 to Feb. 2020 were enrolled and randomized to trabeculectomy group(50 eyes of 50 cases)or EX-PRESS drain implantation group(52 eyes of 52 cases)3-5d after conbercept injections. The concentrations of VEGF, IL-6 and IL-8 in aqueous humor were determined by enzyme-linked immunosorbent assay(ELISA). The therapeutic efficacies of different surgical modalities were evaluated and compared by status of iris neovascularization, changes in postoperative intraocular pressure(IOP), improvement of visual acuity and incidence of complications.

RESULTS:Decreases in aqueous humor concentrations of VEGF, IL-6 and IL-8 were observed at 3-5d after treatment of conbercept(all P<0.05). At 1, 3d, 1, 3, 6 and 12mo after surgery, the IOP levels of patients in both groups were significantly reduced compared to those before surgery(all P<0.05), and there was a statistically significant difference in IOP between the two groups at 3, 6, and 12mo postoperatively(all P<0.05). At 6 and 12mo after surgery, patients treated with EX-PRESS drain implantation showed better visual acuity compared to patients treated with trabeculectomy(all P<0.05). There was no statistically significant difference in types and dosages of anti-glaucoma drugs administered to patients in different groups. At 12mo follow-up, success rate of surgery in the EX-PRESS drain implantation group(86.5%)was higher than that in the trabeculectomy group(70.0%), along with remarkably lower incidence rate of complications compared to that of the trabeculectomy group(P<0.05).

CONCLUSION: The intravitreal injection of conbercept could down-regulate aqueous humor concentrations of VEGF, IL-6 and IL-8. Both of trabeculectomy and EX-PRESS drain implantation could reduce IOP in NVG patients, but the latter procedure had fewer incidence of complications and was more advantageous in improving visual acuity.