OBJECTIVE: To investigate the biological activity and antitumor effect of pegylated recombinant human interleukin 2 (PEG-rhIL-2) obtained by site-specific conjugation of polyethylene glycol (PEG) with non-natural amino acids, and to explore its antitumor mechanism. METHODS: The binding activities of PEG-rhIL-2 at three different sites (T41, Y45, and V91) to human interleukin 2 receptors α (IL-2R_α) and β (IL-2R_β) and were detected by surface plasmon resonance (SPR) technology. Western blot was used to detect the levels of the Janus kinase-signal transducer and activator of transcription 5 (JAK-STAT5) signaling pathway activated by different doses of rhIL-2 and PEG-rhIL-2 in CTTL-2 and YT cells. Blood was collected after a single administration in mice to detect the drug concentration at different time points and evaluate the pharmacokinetic parameters of Y45-PEG-rhIL-2. Mouse hepatoma cell line Hepa1-6, pancreatic cancer cell line Pan-02, and colon cancer cell line MC-38 were selected. Tumor models were constructed in C57BL/6 mice. Different doses of Y45-PEG-rhIL-2 and excipient control were administrated respectively to evaluate the tumor suppression effect of the drug. In the MC-38 colon cancer model, the tumor suppression effect of Y45-PEG-rhIL-2 combined with anti-programmed death-1 (PD-1) monoclonal antibody was evaluated. Hepa1-6 mouse tumor models were constructed and rhIL-2, Y45-rhIL-2 and Y45-PEG-rhIL-2 were administrated respectively. The proportion of tumor-infiltrating lymphocytes was analyzed by flow cytometry. RESULTS: The SPR detection results showed that the binding activities of PEG-rhIL-2 to IL-2R_α/IL-2R_β were both reduced. The affinity of Y45-PEG-rhIL-2 to IL-2R_α was reduced to approximately 1/250, and its affinity to IL-2R_β was reduced to 1/3. Western blot results showed that the activity of Y45-PEG-rhIL-2 in stimulating JAK-STAT5 signaling in CTLL-2 cells expressing heterotrimeric IL-2 receptor complex IL-2R_αβγwas reduced to approximately 1/300, while its activity in YT cells expressing heterodimeric IL-2 receptor complex IL-2R_βγwas reduced to approximately 1/3. The pharmacokinetic evaluation after a single dose in the mice showed that the elimination half-life of Y45-PEG-rhIL-2 was 17.7 h. Y45-PEG-rhIL-2 has pharmacokinetic characteristics superior to those of rhIL-2. Y45-PEG-rhIL-2 showed dose-dependent tumor suppression activity, and the combination of Y45-PEG-rhIL-2 and anti-PD-1 antibody had a better tumor-inhibiting effect than the single use of Y45-PEG-rhIL-2 or anti-PD-1 antibody. Flow cytometry analysis demonstrated that 72 h after the administration of Y45-PEG-rhIL-2, the proportion of tumor-infiltrating cytotoxic T lymphocytes (CD8⁺T cells) increased by 86.84%. At 120 h after administration, the ratio of CD8⁺T cells to regulatory T cells (Treg) increased by 75.10%. CONCLUSION Y45-PEG-rhIL-2 obtained by site-specific conjugation via non-natural amino acids changed its receptor binding activity and inhibited tumor growth in dose-dependent manner in multiple tumor models by regulating CD8⁺T cells.
Interleukin-2/pharmacokinetics* ; Animals ; Mice ; Humans ; Recombinant Proteins/pharmacology* ; Polyethylene Glycols/chemistry* ; Cell Line, Tumor ; Antineoplastic Agents/pharmacokinetics* ; Signal Transduction/drug effects* ; STAT5 Transcription Factor/metabolism* ; Interleukin-2 Receptor alpha Subunit/metabolism* ; Interleukin-2 Receptor beta Subunit/metabolism*

Intentional tooth replantation (ITR) is an advanced treatment modality and the procedure of last resort for preserving teeth with inaccessible endodontic or resorptive lesions. ITR is defined as the deliberate extraction of a tooth; evaluation of the root surface, endodontic manipulation, and repair; and placement of the tooth back into its original socket. Case reports, case series, cohort studies, and randomized controlled trials have demonstrated the efficacy of ITR in the retention of natural teeth that are untreatable or difficult to manage with root canal treatment or endodontic microsurgery. However, variations in clinical protocols for ITR exist due to the empirical nature of the original protocols and rapid advancements in the field of oral biology and dental materials. This heterogeneity in protocols may cause confusion among dental practitioners; therefore, guidelines and considerations for ITR should be explicated. This expert consensus discusses the biological foundation of ITR, the available clinical protocols and current status of ITR in treating teeth with refractory apical periodontitis or anatomical aberration, and the main complications of this treatment, aiming to refine the clinical management of ITR in accordance with the progress of basic research and clinical studies; the findings suggest that ITR may become a more consistent evidence-based option in dental treatment.
Humans ; Tooth Replantation/methods* ; Consensus ; Periapical Periodontitis/surgery*

This paper summarizes the experience of professor DING Yuanqing in treating post-stroke depression （PSD） of young and middle-aged patients with the method of regulating qi and promoting blood circulation. PSD is a syndrome resulting by vascular injury and impairment of brain marrow and vital activity after the stroke. Factors such as poor lifestyle， improper control of chronic diseases and sleep disorders，etc.，which can be harmful individually， or they can interact. Over time，these factors can block yang of defensive qi，obstract blood circulationg and disturb qi movement. Reverse ascending of defensive qi can generate wind and fire，generate phlegm and stasis from the fluid the blood. Qi stagnation， phlegm and stasis can combined with stagnation heat， phlegm heat， blood stasis heat which caused by stroke ， which can further aggravate pulse accumulation， damage the blood vessels and block the collaterals. Consequently， defensive qi is floating over and nutrient qi is not smooth， resulting in inadequate nourishment of the brain marrow，and disfunction of vital activity， causing depressive symptoms. Professor DING innovatively applied the method of regulating qi and promoting blood circulation. He selected the classic prescriptions such as Guizhi Decoction（桂枝汤）， Baoyuan Decoction（保元汤）， as well as self-fitting prescriptions like Erdan Decoction（二丹汤）， Erzhu Decoction（二竹汤）， to relieve qi and tonify qi，promote harmonious blood circulation， facilitate vasodilation， ease symptoms of depression， invigorate the mind， and provide an effective treatment for PSD.

【Objective】 To investigate the effect of clinical factors and inflammatory markers on platelet transfusion refractoriness (PTR) in children with chemotherapy- induced thrombocytopenia (CIT) and evaluate their predictive value, so as to provide reference for mechanism study of PTR caused by non-immune factors in children with cancer and provide clinical guidance for reasonable and effective platelet transfusion in children. 【Methods】 A total of 60 CIT pediatric cancer patients from Children’s Hospital, Zhejiang University School of Medicine between November 2022 and February 2024 were selected as the study subjects, and divided into the PTR group (n=30) and the non-PTR group (n=30) to collect the clinical data and laboratory markers before platelet transfusion (Plt). Univariate and multivariate logistic regression analysis was used to analyze the influencing factors of PTR. The predictive value influencing factors of PTR was analyzed by receiver operating characteristic (ROC) curve. 【Results】 Plasma IL-1β concentration in PTR group was significantly higher than that in non-PTR group[67.43 pg/mL (29.38, 222.40) vs 36.38 pg/mL (17.27, 68.06); P<0.05]; plasma IL-8 concentration in PTR group was significantly higher than that in non-PTR group[60.97 pg/mL (39.07, 112.00) vs 25.23 pg/mL (5.00, 71.38); P<0.01]; and the interval from initiation of chemotherapy to platelet transfusion was significantly higher in the PTR group than in the non-PTR group[9.5 d (8.0, 12.0) vs 12.0 d (9.8, 13.2); P<0.05]. Multivariate logistic regression analysis showed that IL-8 concentration (OR=1.05, P<0.05) had a statistically significant effect on the occurrence of PTR. ROC curve analysis showed that plasma IL-1β concentration (Cut-off value: 64.88 pg/mL; AUC: 0.653[95% CI: 0.511-0.796]) and plasma IL-8 concentration (Cut-off value: 33.33 pg/mL; AUC: 0.754[95% CI: 0.631-0.878]) and the interval from initiation of chemotherapy to platelet transfusion (Cut-off value: 11.5 days; AUC: 0.669[95% CI: 0.529-0.810]) were statistically significant in predicting PTR. 【Conclusion】 Plasma IL-8 concentration is an independent risk factor for PTR, and plasma IL-1β, IL-8 concentration and interval from initiation of chemotherapy to platelet transfusion have predictive value for PTR.

Objective:To discuss the effects of leucovorin on tumor growth and angiogenesis in the mice with lung cancer transplantation tumor,and to elucidate its mechanism.Methods:Thirty-two healthy C57BL/6 mice were selected to prepare the Lewis lung cancer transplantation tumor models.The mice were randomly divided into model group(0.9%NaCl),low dose of leucovrin group(25 mg·kg-1 leucovrin),high dose of leucovrin group(50 mg·kg-1 leucovrin)and positive control group(60 mg·kg-1 cyclophosphamide),and there were 8 mice in each group.The levels of interleukin-2(IL-2),interferon-γ(INF-γ)and tumor necrosis factor-α(TNF-α)in serum of the mice in various groups were detected by enzyme linked immunosorbent assay(ELISA)method;the expression of vascular endothelial growth factor(VEGF)in tumor tissue of the mice in various groups were detected by immunohistochemistry,and the microvascular density(MVD)and VEGF protein expression scores were calculated;the expression levels of nuclear factor-κB(NF-κB)and hypoxia inducible factor-1α(HIF-1α)in tumor tissue of the mice in various groups were detected by Western blotting method.Results:Compared with model group,the transplantation tumor masses of the mice in low and high doses of leucovorin groups and positive control group were decreased(P<0.05);compared with low dose of leucovorin group,the transplantation tumor masses of the mice in high dose of leucovorin group and positive control group were decreased(P<0.05),and the tumor inhibitory rates were increased(P<0.05);compared with high dose of leucovorin group,the transplantation tumor mass of the mice in positive control group was decreased(P<0.05),and the tumor inhibitory rate was increased(P<0.05).Compared with model group,the spleen indexes and thymus indexes of the mice in low and high doses of leucovorin groups and positive control group were increased(P<0.05);compared with low dose of leucovorin group,the spleen indexes and thymus indexes of the mice in high dose of leucovorin group and positive control group were increased(P<0.05);compared with high dose of leucovorin group,the spleen index and thymus index of the mice in positive control group were increased(P<0.05).The ELISA method results showed that compared with model group,the levels of IL-2,INF-γ and TNF-α in serum of the mice in low and high doses of leucovorin groups and positive control group were increased(P<0.05);compared with low dose of leucovorin group,the levels of IL-2,INF-γ and TNF-α in serum of the mice in high dose of leucovorin group and positive control group were increased(P<0.05);compared with high dose of leucovorin group,the levels of IL-2,INF-γ and TNF-α in serum of the mice in positive control group were increased(P<0.05).Compared with model group,the MVD in tumor tissue of the mice in low and high dose of leucovorin groups and positive control group was decreased(P<0.05);compared with low dose of leucovorin group,the MVD in tumor tissue of the mice in high dose of leucovorin group and positive control group was decreased(P<0.05);compared with high dose of leucovorin group,the MVD in tumor tissue of the mice in positive control group was decreased(P<0.05).Compared with model group,the expression amounts of VEGF protein in tumor tissue of the mice in low and high dose of leucovorin groups and positive control group were decreased;compared with low dose of leucovorin group,the expression amounts of VEGF protein in tumor tissue of the mice in high dose of leucovorin group and positive control group were decreased;compared with high dose of leucovorin group,the expression amount of VEGF protein in tumor tissue of the mice in positive control group was decreased.The differences in VEGF protein expression scores between model group,low dose of leucovorin group,high dose of leucovorin group and positive control group were statistically significant(P<0.05).Compared with model group,the expression levels of NF-κB and HIF-1α in tumor tissue of the mice in low and high doses of leucovorin groups and positive control group were decreased(P<0.05);compared with low dose of leucovorin group,the expression levels of NF-κB and HIF-1α in tumor tissue of the mice in high dose of leucovorin group and positive control group were decreased(P<0.05);compared with high dose of leucovorin group,the expression levels of NF-κB and HIF-1 α in tumor tissue of the mice in positive control group were decreased(P<0.05).Conclusion:Leucolorin can inhibit the tumor growth,protect the immune organs and suppress the tumor angiogenesis in the Lewis lung cancer transplantation tumor mice,and may exert the therapeutic effect by targeting NF-κB/HIF-1α signaling pathway and down-regulating the expression levels of NF-κB and HIF-1α proteins.

Objective:To investigate the clinical efficacy of antiperistaltic uncut Roux-en-Y anastomosis (Uncut RY) in laparoscopic distal gastrectomy (LDG) for gastric cancer (GC).Methods:The clinicpathological data of 309 patients who underwent LDG for GC between Jan 2020 to Dec 2021 in Tianjin Medical University Cancer Institute & Hospital were collected. Among them, 102 cases received antiperistaltic Uncut RY anastomosis, 96 cases did RY anastomosis, and 111 cases did B Ⅱ+Braun anastomosis. The general data and clinical efficacy were compared.Results:The operative time in Uncut RY group and B Ⅱ+Braun group was shorter than that in RY group ( F=3.356, P=0.036). The incidence of postoperative short-term complications was not statistically different among three groups ( P>0.05). Compared to Uncut RY group and RY group, the B Ⅱ+Braun group had a significantly higher incidences of reflux gastritis (19.8% vs. 6.9% vs.5.2%, F=13.889, P=0.001). The incidence of RY stasis syndrome in RY group (15.6%) was significantly higher than that in Uncut RY group (2.0%) and B Ⅱ+Braun group (2.7%) ( F=19.320 ,P=0.000). There were no significant differences in postoperative 1-year nutritional indexes among three group ( P>0.05). Conclusions:The antiperistaltic Uncut RY anastomosis in LDG for GC is safe and feasible, which can reduce the incidences of reflux gastritis and RY stasis syndrome.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective To clarify the changes in CXCL5 in serum and gastric tissues of patients with chronic atrophic gastritis(CAG)and precancerous lesions of gastric cancer(PLGC)and to investigate the predictive value of CXCL5 for the diagnosis of CAG and PLGC.Methods This study enrolled 72 participants of CAG admitted to the Department of Splenology and Gastroenterology of the Second Affiliated Hospital of Anhui University of Chinese Medicine from June 2022 to June 2023,with gastroscopy and pathologically confirmed diagnosis,as well as 68 healthy participants who underwent gastroscopy in the same period at our department.We collected clinical information and laboratory results from all participants.The logistic regression analysis methods were used to identify the diagnostic value of serum CXCL5.Furthermore,in order to clarify the role of CXCL5 in the developent of CAG,a total of 15 patients each with CAG,intestinal metaplasia,and dysplasia treated at our hospital from June 2023 to December 2023,and 15 healthy participants were selected.The relationship between the expression of CXCL5 and the degree of clinicopathology was analysed in each group using ELISA,PCR,and immunohistochemistry staining to validate and assess the diagnostic efficacy of CXCL5.Results The study found that several factors were associated with CAG,including family history of tumours,smoking and alcohol consumption history,dietary regularity,Helicobacter pylori infection,the number of lesions,gastric function scores and CXCL5(P<0.05).The ROC curve had an AUC of 1.00 and a Youden index of 0.986,indicating excellent predictive ability.The ELISA results indicated a significantly higher serum CXCL5 expression level in the CAG,intestinal metaplasia,and dysplasia groups compared to the normal group.There was a positive correlation between the serum CXCL5 expression level and the degree of pathology.The PCR and immunohistochemistry staining results indicate that the mRNA and pro-tein expression levels of CXCL5 in gastric tissues of patient groups were significantly higher compared to the nor-mal group.Furthermore,the mRNA and protein expression levels of CXCL5 in gastric tissues were positively correlated with the degree of pathology.Conclusions The results indicate that CXCL5 is highly expressed in the serum and gastric tissues of patients with CAG and PLGC,and its expression level is positively correlated with the degree of pathology.Therefore,CXCL5 could serve as a predictive indicator and a potential therapeutic target for the diagnosis of CAG and PLGC.

The dental operative microscope has been widely employed in the field of dentistry, particularly in endodontics and operative dentistry, resulting in significant advancements in the effectiveness of root canal therapy, endodontic surgery, and dental restoration. However, the improper use of this microscope continues to be common in clinical settings, primarily due to operators' insufficient understanding and proficiency in both the features and established operating procedures of this equipment. In October 2019, Professor Jingping Liang, Vice Chairman of the Society of Cariology and Endodontology, Chinese Stomatological Association, organized a consensus meeting with Chinese experts in endodontics and operative dentistry. The objective of this meeting was to establish a standard operation procedure for the dental operative microscope. Subsequently, a consensus was reached and officially issued. Over the span of about four years, the content of this consensus has been further developed and improved through practical experience.
Humans ; Dentistry, Operative ; Consensus ; Endodontics ; Root Canal Therapy ; Dental Care

Objective:To analyze the individual iodine nutrition status and its influencing factors among students aged 8 to 15 in Dongtai City, Jiangsu Province.Methods:From May to August 2021, a total of 905 students aged 8 to 15 were selected as survey subjects in Dongtai City based on the sampling method in the "National Monitoring Plan for Iodine Deficiency Disorders" (2016 version). Salt samples from students' homes were collected for salt iodine testing. Urine samples of students were collected for urinary iodine and creatinine testing. The creatinine correction method was used to estimate individual 24 h urinary iodine excretion and calculate iodine intake. At the same time, basic information (age, gender, height, weight, etc.) and consumption frequency of iodine rich foods (seafood, eggs, meat, milk, solid snacks) of students were collected through questionnaires and actual measurements.Results:The coverage rate of iodized salt in Dongtai City was 98.2% (889/905), the qualified rate of iodized salt was 97.9% (870/889), and the consumption rate of qualified iodized salt was 96.1% (870/905). The median estimated 24 h urinary iodine excretion was 179.7 μg/d. The median estimated iodine intake was 195.4 μg/d, the constituent ratio of estimated iodine intake < recommended nutrient intake (RNI) was 16.2% (147/905), RNI-tolerable upper intake level (UL) was 63.4% (574/905), and > UL was 20.3% (184/905). The medians estimated 24 h urinary iodine excretion of students aged 8-10, 11-13 and 14-15 were 157.4, 193.0 and 236.5 μg/d, respectively, and the difference was statistically significant ( H = 55.42, P < 0.001). The median estimated 24 h urinary iodine excretion of boys was higher than that of girls (222.6 vs 148.6 μg/d), and the median estimated 24 h urinary iodine excretion of urban students was higher than that of township students (215.6 vs 162.7 μg/d), the differences were statistically significant ( Z = - 8.41, - 5.66, P < 0.001). There were statistically significant differences in the median estimated 24 h urinary iodine excretion between students with different body mass index (weight loss, overweight, obesity, normal; H = 56.15, P < 0.001) and iodine rich foods consumption frequencies (seafood, meat, milk, eggs, solid snacks; H = 23.15, 21.20, 60.77, 20.01, 24.47, P < 0.001). Conclusion:Iodine deficiency or excess exists in students aged 8-15 in Dongtai City, and girls aged 8-10 who are physically emaciated are the focus of attention for iodine deficiency.