1.Strategies and advances in hepatitis B vaccination in China
Lin TANG ; Xia XU ; Zemei ZHOU ; Xiaoqi WANG ; Jizhen LI ; Zundong YIN ; Fuzhen WANG
Journal of Clinical Hepatology 2025;41(2):210-215
Hepatitis B is a major global public health issue. Through the implementation of comprehensive prevention and control strategies centered on hepatitis B vaccination, China has achieved remarkable progress in hepatitis B prevention and control, while there are still many issues and challenges. This article reviews the development of hepatitis B vaccination strategies in China, analyzes the goal and advances in vaccination in different populations, and problems and challenges, in order to provide a reference for further optimizing vaccination strategies and improving the levels of prevention and control.
2.Effect of Icariin on Steroid-induced Ferroptosis in Rat Bone Microvascular Endothelial Cells
Jiancheng TANG ; Yue ZHANG ; Ruichen JIANG ; Zhengrong YUE ; Ming LI ; Yaqi ZHANG ; Zetao YIN ; Weiguo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):131-140
ObjectiveTo investigate the effect of icariin (ICA) on steroid-induced ferroptosis in bone microvascular endothelial cells (BMECs). MethodsRat BMECs were selected and treated with 500 mg·L-1 hydrocortisone for 1.5 h to establish a ferroptosis model of BMECs. The experimental cells were divided into a blank group, hormone group (500 mg·L-1 hydrocortisone), ICA group (500 mg·L-1 hydrocortisone + 34 mg·L-1 ICA), and ferroptosis agonist group (500 mg·L-1 hydrocortisone + 34 mg·L-1 ICA + 2.7 mg·L-1 erastin). Cell viability was detected by CCK-8. The levels of ferrous ion, glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and reactive oxygen species (ROS) were detected by related kit species. The ferroptosis-related proteins, such as glutathione peroxidase 4(GPX4), ferritin light chain (FTL), and transferrin receptor protein1 (sTfR) were detected by Western blot, as well as autophagy-related proteins including microtubule-associated protein 1 light chain 3B (LC3B), Beclin1, B-cell lymphoma-2 (Bcl-2), and Caspase-3. Results500 mg·L-1 hydrocortisone intervention for 1.5 h could effectively induce ferroptosis in BMECs, and ferroptosis levels could reach a peak as the intervention continued. In terms of cellular antioxidant capacity, compared with those in the blank group, the cell vitality, GSH in the hormone group decreased significantly, and the levels of ROS, SOD, MDA, and ferrous ions were significantly increased (P<0.01). Compared with those in the hormone group, the cell viability, GSH were significantly increased, and the levels of ROS, SOD, MDA, and ferrous ions were decreased in the ICA group (P<0.01). Compared with those in the ICA group, the cell vitality, GSH in the ferroptosis agonist group decreased significantly, and the levels of ROS, SOD, MDA, and ferrous ions increased significantly (P<0.01). In terms of the relationship between ferroptosis and autophagy, compared with the blank group, the hormone group had significantly increased expression levels of LC3B, sTfR, Beclin1, and FTL and significantly decreased expression levels of GPX4 (P<0.01). Compared with the hormone group, The ICA group had significantly decreased expression levels of LC3B, sTfR, and FTL and significantly increased expression levels of Beclin 1 and GPX4 (P<0.01). Compared with those in the ICA group, the expression levels of LC3B, sTfR, and FTL increased in the rapamycin group, and those of Beclin 1 and GPX4 decreased (P<0.01). In terms of cell ferroptosis and apoptosis,compared with the blank group, the hormone group had significantly increased expression levels of FTL, sTfR and Caspase-3 and significantly decreased expression levels of GPX4, and Bcl-2 (P<0.01). Compared with the hormone group, the ICA group had significantly decreased expression levels of FTL, sTfR and Caspase-3 and significantly increased expression levels of GPX4, and Bcl-2 (P<0.01). Compared with those in the ICA group, the expression levels of FTL, sTfR and Caspase-3 in the ferroptosis agonist group were increased, and the expression levels of GPX4, and Bcl-2 were decreased (P<0.01). In terms of cell function,compared with that in the blank group, the ability of cell migration and tube formation was significantly decreased in the hormone group (P<0.01). Compared with that in the hormone group, the cell migration and tube formation ability in the ICA group were significantly increased (P<0.01). ConclusionFerroptosis is involved in steroid-induced damage in BMECs. ICA can inhibit steroid-induced ferroptosis in BMECs, and the mechanism may be associated with the inhibition of ferroptosis by regulating autophagy.
3.Preparation and evaluation of quality,targeting and cytotoxicity of triptolide-loaded targeting nanoparticles
Moli YIN ; Wenbin LUO ; Jingzhe XU ; Zebo TANG ; Ni GUO ; Youxing LAO ; Huiyan WANG
China Pharmacy 2025;36(12):1457-1462
OBJECTIVE To prepare nanoparticle-based targeting preparation loaded with triptolide (TP), and evaluate its quality, targeting ability and cytotoxic effects. METHODS Polymer nanoparticles carrying TP-targeted folic acid (FA) receptor (TP@PLGA-PEG-FA) were fabricated using poly (lactic-co-glycolic acid)/polyethylene glycol/FA (PLGA-PEG-FA) as the carrier by emulsion and volatilization technique. The morphology and distribution were observed, and their particle size, Zeta potential, polydispersity index (PDI), drug loading capacity and encapsulation efficiency were measured. Their stability, blood compatibility, in vitro drug release, uptake by RAW264.7 cells (localization with fluorescent dye Cy3.5), and in vitro cytotoxicity were evaluated. RESULTS TP@PLGA-PEG-FA exhibited spherical shape and uniform distribution, with particle size of (122.60±0.02) nm, Zeta potential of (-17.6±0.6)mV, and PDI of 0.26±0.02; drug loading capacity and encapsulation efficiency of TP were measured to be (7.78±0.05)% and (68.62±0.03)%, respectively. The hemolysis rates of 100, 200, 300, 400 µg/mL TP@PLGA- PEG-FA were 0.77%, 0.92%, 1.34% and 1.63%, respectively. There were no significant changes in particle size, PDI and Zeta potential when TP@PLGA-PEG-FA were placed in 4 ℃ water for 14 days and in DMEM culture medium containing 10% fetal bovine serum at 37 ℃ for 12 h. The cumulative release rate of TP@PLGA-PEG-FA was (84.83±0.29)% in phosphate buffer at pH5.5 for 72 h, which was significantly higher than the cumulative release rates in phosphate buffer solutions at pH7.4 and 6.5 for 72 h ([ 42.37±0.35)% and (63.83±0.29)% , respectively] (P<0.05). Activated RAW264.7 cells took up significantly more Cy3.5@PLGA-PEG-FA than they took up Cy3.5@PLGA-PEG-FA+free FA and Cy3.5@PLGA-PEG. When the mass concentration of TP was≥15.63 ng/mL, the survival rates of activated cells in the TP@PLGA-PEG-FA groups were significantly lower than those of the same mass concentration of free TP groups (P<0.05). CONCLUSIONS The prepared TP@PLGA-PEG-FA has high stability, good blood compatibility, active targeting and cytotoxicity to inflammatory cells.
4.Study on Kinetic and Static Tasks With Different Resistance Coefficients in Post-stroke Rehabilitation Training Based on Functional Near-infrared Spectroscopy
Ling-Di FU ; Jia-Xuan DOU ; Ting-Ting YING ; Li-Yong YIN ; Min TANG ; Zhen-Hu LIANG
Progress in Biochemistry and Biophysics 2025;52(7):1890-1903
ObjectiveFunctional near-infrared spectroscopy (fNIRS), a novel non-invasive technique for monitoring cerebral activity, can be integrated with upper limb rehabilitation robots to facilitate the real-time assessment of neurological rehabilitation outcomes. The rehabilitation robot is designed with 3 training modes: passive, active, and resistance. Among these, the resistance mode has been demonstrated to yield superior rehabilitative outcomes for patients with a certain level of muscle strength. The control modes in the resistance mode can be categorized into dynamic and static control. However, the effects of different control modes in the resistance mode on the motor function of patients with upper limb hemiplegia in stroke remain unclear. Furthermore, the effects of force, an important parameter of different control modes, on the activation of brain regions have rarely been reported. This study investigates the effects of dynamic and static resistance modes under varying resistance levels on cerebral functional alterations during motor rehabilitation in post-stroke patients. MethodsA cohort of 20 stroke patients with upper limb dysfunction was enrolled in the study, completing preparatory adaptive training followed by 3 intensity-level tasks across 2 motor paradigms. The bilateral prefrontal cortices (PFC), bilateral primary motor cortices (M1), bilateral primary somatosensory cortices (S1), and bilateral premotor and supplementary motor cortices (PM) were examined in both the resting and motor training states. The lateralization index (LI), phase locking value (PLV), network metrics were employed to examine cortical activation patterns and topological properties of brain connectivity. ResultsThe data indicated that both dynamic and static modes resulted in significantly greater activation of the contralateral M1 area and the ipsilateral PM area when compared to the resting state. The static patterns demonstrated a more pronounced activation in the contralateral M1 in comparison to the dynamic patterns. The results of brain network analysis revealed significant differences between the dynamic and resting states in the contralateral PFC area and contralateral M1 area (F=4.709, P=0.038), as well as in the contralateral PM area and ipsilateral M1 area (F=4.218, P=0.049). Moreover, the findings indicated a positive correlation between the activation of the M1 region and the increase in force in the dynamic mode, which was reversed in the static mode. ConclusionBoth dynamic and static resistance training modes have been demonstrated to activate the corresponding brain functional regions. Dynamic resistance modes elicit greater oxygen changes and connectivity to the region of interest (ROI) than static resistance modes. Furthermore, the effects of increasing force differ between the two modes. In patients who have suffered a stroke, dynamic modes may have a more pronounced effect on the activation of exercise-related functional brain regions.
5.Induction of apoptosis in hepatocellular carcinoma cells by polyphyllin 9 through regulating the Fas/FasL sig-naling pathway and the inhibitory effect on the growth of transplanted tumor in nude mice
Minna YAO ; Wei ZHANG ; Kai GAO ; Ruili LI ; Ying YIN ; Chao GUO ; Yunyang LU ; Haifeng TANG ; Jingwen WANG
China Pharmacy 2025;36(18):2238-2243
OBJECTIVE To investigate the induction of apoptosis in hepatocellular carcinoma cells by polyphyllin 9 (PP9) through the regulation of the Fas/Fas ligand (FasL) signaling pathway, and its inhibitory effect on the growth of transplanted tumor in nude mice. METHODS Based on the screening of cell lines and intervention conditions, HepG2 cells were selected as the experimental subject to investigate the effects of 2 μmol/L and 4 μmol/L PP9 treatment on cell colony formation activity, apoptosis rate, as well as the protein expressions of Fas, FasL, cleaved caspase-8 and cleaved caspase-3. Additionally, Fas inhibitor KR- 33493 was introduced to investigate the underlying mechanism of PP9’s anti-hepatocellular carcinoma activity. Using HepG2 cell tumor-bearing nude mice model as the object, and 5-fluorouracil (20 mg/kg) as the positive control, the effects of 10 mg/kg PP9 on tumor volume, tumor mass, and the protein expressions of the nuclear proliferation-associated antigen Ki-67 and cleaved caspase-3 in tumor-bearing nude mice were investigated. RESULTS Compared with the control group, 2, 4 μmol/L PP9 significantly decreased the number of clones and the clone formation rate of cells, but significantly increased the apoptosis rate, the protein expressions of Fas, FasL, cleaved caspase-8 and cleaved caspase-3 (P<0.05 or P<0.01). However, the combination of Fas inhibitor KR-33493 could significantly reverse the effect of PP9 on the up-regulation of proteins related to the Fas/FasL signaling pathway (P<0.01). Compared with the control group, the tumor volume (on day 27), mass and protein expression of Ki- 67 in nude mice of the PP9 group were significantly decreased, while the protein expression of cleaved caspase-3 was significantly increased (P<0.01). CONCLUSIONS PP9 can induce apoptosis of HepG2 cells by activating the Fas/FasL signaling pathway. Meanwhile, PP9 can also effectively inhibit the growth of transplanted tumors in nude mice.
6.Development and application of core competency evaluation index system for ethics committee members of drug clinical trial institution
Xiaohua TANG ; Qingqing JIANG ; Haiwei ZHANG ; Weiran MAO ; Yuanyuan YIN ; Jiangdong SUI ; Jun LI ; Xia CHEN
China Pharmacy 2025;36(20):2489-2494
OBJECTIVE To construct an evaluation index system for the core competencies of ethics committee members of drug clinical trial institution, providing a basis for optimizing the training system for committee members, improving the quality of ethical review, and fully safeguarding the safety and rights of subjects. METHODS Using methods such as literature research and expert consultation, a preliminary core competency evaluation index system was constructed. The Delphi method was employed to revise and validate it, ultimately forming an evaluation index system for the core competencies of ethics committee members. Based on this system, a questionnaire survey was conducted among 90 ethics committee members from 29 drug clinical trial institutions nationwide, comparing their importance rating and self-assessment scores of the core competency indexes. RESULTS The evaluation system constructed included 4 primary indicators (ethics and professional knowledge, ethics review ability, communication and expression ability, moral integrity and work style) and 39 secondary indicators (familiarity with the content of clinical trial-related laws and regulations, ability to complete project ethics review and identify ethical defects in research protocols within a short period of time, ability to judge the scientific value of clinical research, etc.). The results of questionnaire survey showed that the interviewed ethics committee members had significant capability gaps in dimensions such as regulatory knowledge, ethical norms, review efficiency, risk judgment, and problem analysis. The differences between the importance rating scores of corresponding secondary indicators and the self-assessment scores were all no less than 0.38. CONCLUSIONS This study has developed a quantifiable and stratified core competency assessment tool for ethics committee members. It can provide a scientific framework for committee member training, qualification certification, and standardized management of ethics committees.
7.Study on the Construction of a Question-Answer Corpus Dataset for Chinese Medical Knowledge Large Language Models
Tingyu LYU ; Xiaoying LI ; Ying ZHANG ; Yuyang LIU ; Jinhua DU ; Xinyi LI ; Yan LUO ; Xiaoli TANG ; Huiling REN ; Hui LIU ; Hao YIN
Journal of Medical Informatics 2024;45(5):20-25
Purpose/Significance To construct a Chinese medical knowledge Q&A corpus dataset as a standardized evaluation bench-mark for large language models(LLMs)in the medical domain,so as to improve the accuracy and efficiency of LLMs in handling Chinese medical questions.Method/Process Chinese medical paper knowledge,medical terminology explanations and supplementary questions are acquired from the Chinese medical licensing examination,and open-source Chinese medical Q&A datasets are encompassed in the developed Q&A datasets.Result/Conclusion The Chinese medical knowledge Q&A corpus datasets enrich the sources of existing datasets and promote the objective and comprehensive quantitative evaluation of large models in the medical field.In the near future,additional data such as electronic medical records and those from online health communities will be used to strengthen the support of artificial intelli-gence for the Healthy China strategy.
8.Distribution and antimicrobial resistance of bacterial strains isolated from blood samples in a traditional Chinese medicine hospital in Shenzhen
Xutao ZHENG ; Rimei ZHANG ; Qiong DUAN ; Shanru LIN ; Jialing TANG ; Lingfan YIN
Chinese Journal of Infection and Chemotherapy 2024;24(4):442-447
Objective To investigate the distribution and antimicrobial resistance of the bacterial strains isolated from blood samples of inpatients in Longgang Hospital,Beijing University of Chinese Medicine.Methods The bacterial identification and antimicrobial susceptibility test results for the strains isolated from 2018 to 2022 were retrospectively analyzed.Results A total of 910 strains of bacteria were isolated from blood samples,of which 63.2%(575/910)were gram-negative bacteria and 36.8%(335/910)were gram-positive bacteria.Escherichia coli,coagulase-negative Staphylococcus,Klebsiella pneumoniae,Staphylococcus aureus,and Enterococcus spp.were the top 5 pathogens.In the past 5 years,no carbapenem-resistant strains of E.coli or K.pneumoniae were found in the blood samples of the inpatients.A.baumannii had a resistance rate of 11.8%to carbapenems.The prevalence of methicillin-resistant strains in S.aureus,S.epidermidis and other Staphylococcus species was 16.7%,75.0%and 55.5%,respectively.No vancomycin-or linezolid-resistant staphylococcual isolates were found.No strains of Enterococcus faecalis or Enterococcus faecium were found resistant to high concentrations of gentamicin,linezolid,or vancomycin.Conclusions The bacteria isolated from blood samples in Longgang Hospital were mainly gram-negative bacteria.Carbapenem-resistant strain was identified in the strains of A.baumannii.Bacterial resistance surveillance should be strengthened for the isolates from blood samples and other specimens from the site of infection.Antimicrobial agents should be used rationally to prevent the spread of drug-resistant bacteria.
9.Multicenter retrospect analysis of early clinical features and analysis of risk factors on prognosis of elderly patients with severe burns
Qimin MA ; Wenbin TANG ; Xiaojian LI ; Fei CHANG ; Xi YIN ; Zhaohong CHEN ; Guohua WU ; Chengde XIA ; Xiaoliang LI ; Deyun WANG ; Zhigang CHU ; Yi ZHANG ; Lei WANG ; Choulang WU ; Yalin TONG ; Pei CUI ; Guanghua GUO ; Zhihao ZHU ; Shengyu HUANG ; Liu CHANG ; Rui LIU ; Yongji LIU ; Yusong WANG ; Xiaobin LIU ; Tuo SHEN ; Feng ZHU
Chinese Journal of Burns 2024;40(3):249-257
Objective:To investigate the early clinical characteristics of elderly patients with severe burns and the risk factors on prognosis.Methods:This study was a retrospective case series study. Clinical data of 124 elderly patients with severe burns who met the inclusion criteria and were admitted to the 12 hospitals from January 2015 to December 2020 were collected, including 4 patients from the Fourth People's Hospital of Dalian, 5 patients from Fujian Medical University Union Hospital, 22 patients from Guangzhou Red Cross Hospital of Jinan University, 5 patients from Heilongjiang Provincial Hospital, 27 patients from the First Affiliated Hospital of Naval Medical University, 9 patients from the First Affiliated Hospital of Nanchang University, 10 patients from Affiliated Hospital of Nantong University, 9 patients from Tongren Hospital of Wuhan University & Wuhan Third Hospital, 12 patients from the 924 th Hospital of PLA, 6 patients from Zhangjiagang First People's Hospital, 4 patients from Taizhou Hospital of Zhejiang Province, and 11 patients from Zhengzhou First People's Hospital. The patients' overall clinical characteristics, such as gender, age, body mass index, total burn area, full-thickness burn area, inhalation injury, causative factors, whether combined with underlying medical diseases, and admission time after injury were recorded. According to the survival outcome within 28 days after injury, the patients were divided into survival group (89 cases) and death group (35 cases). The following data of patients were compared between the two groups, including the basic data and injuries (the same as the overall clinical characteristics ahead); the coagulation indexes within the first 24 hours of injury such as prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time, D-dimer, fibrinogen degradation product (FDP), international normalized ratio (INR), and fibrinogen; the blood routine indexes within the first 24 hours of injury such as white blood cell count, platelet count, neutrophil-to-lymphocyte ratio, monocyte count, red blood cell count, hemoglobin, and hematocrit; the organ function indexes within the first 24 hours of injury such as direct bilirubin, total bilirubin, urea, serum creatinine, aspartate aminotransferase, alanine aminotransferase, total protein, albumin, globulin, blood glucose, triglyceride, total cholesterol, alkaline phosphatase, creatine kinase, electrolyte indexes (potassium, sodium, chlorine, calcium, magnesium, and phosphorus in blood), uric acid, myoglobin, and brain natriuretic peptide; the infection and blood gas indexes within the first 24 hours of injury such as procalcitonin, C-reactive protein, pH value, oxygenation index, base excess, and lactate; treatment such as whether conducted with mechanical ventilation, whether conducted with continuous renal replacement therapy, whether conducted with anticoagulation therapy, whether applied with vasoactive drugs, and fluid resuscitation. The analysis was conducted to screen the independent risk factors for the mortality within 28 days after injury in elderly patients with severe burns. Results:Among 124 patients, there were 82 males and 42 females, aged 60-97 years, with body mass index of 23.44 (21.09, 25.95) kg/m 2, total burn area of 54.00% (42.00%, 75.00%) total body surface area (TBSA), and full-thickness burn area of 25.00% (10.00%, 40.00%) TBSA. The patients were mainly combined with moderate to severe inhalation injury and caused by flame burns. There were 43 cases with underlying medical diseases. The majority of patients were admitted to the hospital within 8 hours after injury. There were statistically significant differences between patients in the 2 groups in terms of age, total burn area, full-thickness burn area, and inhalation injury, and PT, APTT, D-dimer, FDP, INR, white blood cell count, platelet count, urea, serum creatinine, blood glucose, blood sodium, uric acid, myoglobin, and urine volume within the first 24 hours of injury (with Z values of 2.37, 5.49, 5.26, 5.97, 2.18, 1.95, 2.68, 2.68, 2.51, 2.82, 2.14, 3.40, 5.31, 3.41, 2.35, 3.81, 2.16, and -3.82, respectively, P<0.05); there were statistically significant differences between two groups of patients in whether conducted with mechanical ventilation and whether applied with vasoactive drugs (with χ2 values of 9.44 and 28.50, respectively, P<0.05). Age, total burn area, full-thickness burn area, serum creatinine within the first 24 hours of injury, and APTT within the first 24 hours of injury were the independent risk factors for the mortality within 28 days after injury in elderly patients with severe burns (with odds ratios of 1.17, 1.10, 1.10, 1.09, and 1.27, 95% confidence intervals of 1.03-1.40, 1.04-1.21, 1.05-1.19, 1.05-1.17, and 1.07-1.69, respectively, P<0.05). Conclusions:The elderly patients with severe burns had the injuries mainly from flame burns, often accompanied by moderate to severe inhalation injury and enhanced inflammatory response, elevated blood glucose levels, activated fibrinolysis, and impaired organ function in the early stage, which are associated with their prognosis. Age, total burn area, full-thickness burn area, and serum creatinine and APTT within the first 24 hours of injury are the independent risk factors for death within 28 days after injury in this population.
10.Mechanism of Tumor T Cell Exhaustion from Perspective of ''Sanjiao-Yingwei'' Qi Transformation Malfunction
Xinhao TANG ; Bowen CHU ; Yuanyuan QIN ; Yeling LIU ; Xinyan SHU ; Mianhua WU ; Gang YIN ; Jianguo DAI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):176-185
In order to promote the innovative application of Sanjiao theory and Yingwei theory, this paper tries to apply the ''Sanjiao-Yingwei'' Qi transformation theory to the treatment of tumor diseases, integrating it with T cell exhaustion mechanism to elaborate on its scientific connotation and using network pharmacology and bioinformatics to elucidate the correlation between the anti-tumor mechanism of ''Sanjiao-Yingwei'' Qi transformation and T cell exhaustion. The ''Sanjiao-Yingwei'' Qi transformation function is closely related to the immunometabolic ability of the human body, and the ''Sanjiao-Yingwei'' Qi transformation system constitutes the immunometabolic exchange system within and outside the cellular environment. Cancer toxicity is generated by the fuzzy Sanjiao Qi, and the long-term fuzzy Sanjiao Qi is the primary factor leading to T cell exhaustion, which is related to the long-term activation of T cell receptors by the high tumor antigen load in the tumor microenvironment. Qi transformation malfunction of the Sanjiao produces phlegm and collects stasis, which contributes to T cell exhaustion and is correlated with nutrient deprivation, lipid accumulation, and high lactate levels in the immunosuppressed tumor microenvironment, as well as with the release of transforming growth factor-β and upregulated expression of programmed death receptor-1 by tumor-associated fibroblasts and platelets in the tumor microenvironment. Ying and Wei damage due to Sanjiao Qi transformation malfunction is similar to the abnormal manifestations such as progressive loss of exhausted T cell effector function and disturbance of cellular energy metabolism. Guizhi decoction, Shengming decoction, and Wendan decoction can correct T cell exhaustion and exert anti-tumor effects through multi-target and multi-pathways by regulating ''Sanjiao-Yingwei'' Qi transformation, and hypoxia inducible factor-1α (HIF-1α) may be one of the main pathways to correct T cell exhaustion. It was found that HIF-1α may be one of the important prognostic indicators in common tumors by bioinformatics. The use of the ''Sanjiao-Yingwei'' Qi transformation method may play an important part in improving the prognosis of tumor patients in clinical practice.

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