To introduce and analyze guideline terminology related to clinical question formulation, evidence retrieval and appraisal, and recommendation development.

OBJECTIVE To provide a reference for standardizing the conduct of positron-emitting radiopharmaceuticals’ phase 0 clinical trials （hereinafter referred to as “phase 0 clinical trials”） and advancing the development of innovative drug by medical institutions. METHODS Initiated by the First Affiliated Hospital of Jinan University， a panel of experts consisting of pharmacy， clinical medicine and medical ethics from multiple institutions was established to investigate the current landscape， and discuss the necessary conditions， procedures， and other aspects for conducting phase 0 clinical trials in medical institutions by integrating relevant national policies， regulations and expert consensus. Finally， an agreement was reached to formulate this consensus. RESULTS & CONCLUSIONS Currently， most medical institutions have deficiencies in pharmaceutical care during the management of radiopharmaceuticals and the phase 0 clinical trials. In conjunction with the Expert Consensus on the Establishment of Nuclear Pharmacist Positions， this consensus explicitly defines the responsibilities of nuclear pharmacists in the phase 0 clinical trials on the basis of the Expert Consensus for the Application of Positron Emission Tomography Radioligands for Translational Study in the Phase 0 Clinical Trials （2020 edition）， providing a guidance for high-quality participation of nuclear pharmacists from medical institutions in China in phase 0 clinical research. Additionally， in consideration of some constraints imposed by current relevant regulations， this consensus also proposes strategic recommendations， such as encouraging medical institutions to form a consortium， leading to the establishment of dedicated bases or industrial parks， holding significant implications to strengthen institutional capacity for advancing radiopharmaceutical innovation through phase 0 clinical trials.

T7 RNA polymerase (T7 RNAP) is one of the simplest known RNA polymerases. Its unique structural features make it a critical model for studying the mechanisms of RNA synthesis. This review systematically examines the static crystal structure of T7 RNAP, beginning with an in-depth examination of its characteristic “thumb”, “palm”, and “finger” domains, which form the classic “right-hand-like” architecture. By detailing these structural elements, this review establishes a foundation for understanding the overall organization of T7 RNAP. This review systematically maps the functional roles of secondary structural elements and their subdomains in transcriptional catalysis, progressively elucidating the fundamental relationships between structure and function. Further, the intrinsic flexibility of T7 RNAP and its applications in research are also discussed. Additionally, the review presents the structural diagrams of the enzyme at different stages of the transcription process, and through these diagrams, it provides a detailed description of the complete transcription process of T7 RNAP. By integrating structural dynamics and kinetics analyses, the review constructs a comprehensive framework that bridges static structure to dynamic processes. Despite its advantages, T7 RNAP has a notable limitation: it generates double-stranded RNA (dsRNA) as a byproduct. The presence of dsRNA not only compromises the purity of mRNA products but also elicits nonspecific immune responses, which pose significant challenges for biotechnological and therapeutic applications. The review provides a detailed exploration of the mechanisms underlying dsRNA formation during T7 RNAP catalysis, reviews current strategies to mitigate this issue, and highlights recent progress in the field. A key focus is the semi-rational design of T7 RNAP mutants engineered to minimize dsRNA generation and enhance catalytic performance. Beyond its role in transcription, T7 RNAP exhibits rapid development and extensive application in fields, including gene editing, biosensing, and mRNA vaccines. This review systematically examines the structure-function relationships of T7 RNAP, elucidates the mechanisms of dsRNA formation, and discusses engineering strategies to optimize its performance. It further explores the engineering optimization and functional expansion of T7 RNAP. Furthermore, this review also addresses the pressing issues that currently need resolution, discusses the major challenges in the practical application of T7 RNAP, and provides an outlook on potential future research directions. In summary, this review provides a comprehensive analysis of T7 RNAP, ranging from its structural architecture to cutting-edge applications. We systematically examine: (1) the characteristic right-hand domains (thumb, palm, fingers) that define its minimalistic structure; (2) the structure-function relationships underlying transcriptional catalysis; and (3) the dynamic transitions during the complete transcription cycle. While highlighting T7 RNAP’s versatility in gene editing, biosensing, and mRNA vaccine production, we critically address its major limitation—dsRNA byproduct formation—and evaluate engineering solutions including semi-rationally designed mutants. By synthesizing current knowledge and identifying key challenges, this work aims to provide novel insights for the development and application of T7 RNAP and to foster further thought and progress in related fields.

AIM: To evaluate the convenience and accuracy of a novel smartphone-assisted “any-point two-step method” for finding the target axial position in cataract phacoemulsification combined with intraocular lens(IOL)implantation.METHODS: Prospective observational study. A total of 62 cases(62 eyes)of patients with age-related cataracts who underwent cataract phacoemulsification combined with IOL implantation in our hospital from October 2021 to April 2022 were selected. They were randomly divided into two groups: 31 cases(31 eyes)in the control group were applied with the “traditional two-step method” using slit lamp to mark the target axial position of the IOL, and 31 cases(31 eyes)in the experimental group were applied with the smartphone-assisted “two-step method” to mark the target axial position of the IOL. The Callisto eye navigation system was used as a standard reference, and the deviation of the reference marking point(deviation-1), the deviation of the target axial marking point(deviation-total), and the deviation of the angle from the reference marking point to the target axial marking point(deviation-2)were calculated and recorded as the preoperative axial marking time.RESULTS:Both deviation-1 and deviation-total values were lower in the experimental group than those in the control group(1.06°±1.39° vs 2.48°±2.23°, 1.77°±1.54° vs 2.81°±1.58°, all P<0.01), but there was no significant difference in the deviation-2 values between the two groups(1.35°±1.40° vs 1.48°±1.79°, P>0.05). The preoperative axial marking took shorter time in the experimental group than in the control group(1.77±1.70 min vs 2.88±3.20 min, P<0.01).CONCLUSION: The smartphone-assisted “any-point two-step method” for finding the target axial position in cataract phacoemulsification combined with IOL implantation is simple, time-saving, and accurate compared with the “traditional two-step method”.

目的：探讨穿心莲内酯（Andro）调节脂肪酸合成酶（Fas）/脂肪酸合成酶配体（FasL）信号轴对子宫内膜癌Ishikawa细胞顺铂（DDP）耐药性的影响。方法：采用0、5、10、20 μg/mL DDP分别处理Ishikawa细胞和顺铂耐药的Ishikawa/DPP细胞，0、5、10、25、50 μmol/L Andro处理Ishikawa/DDP细胞，MTT法检测细胞增殖情况并为后续实验选择合适的给药剂量。将Ishikawa/DDP细胞随机分为对照组、DDP组（DDP干预）、Andro组（DDP、Andro干预）、pcDNA3.1-NC组（转染pcDNA3.1+DDP、Andro干预）、pcDNA3.1-Fas/FasL组（转染pcDNA3.1-Fas/FasL+DDP、Andro干预），24 h后，采用qPCR法检测Fas、FasL mRNA的表达，平板克隆形成实验、Transwell实验和流式细胞术分别检测细胞克隆能力、细胞迁移与侵袭和细胞凋亡，WB法检测增殖细胞核抗原（PCNA）、BAX、Bcl-2、MMP-2、PD-L1、多药耐药蛋白-1（MDR-1）及Fas、FasL蛋白表达。结果：DDP以剂量依赖的方式抑制Ishikawa和Ishikawa/DPP细胞增殖，并且与Ishikawa细胞比较，Ishikawa/DPP细胞对DDP的敏感性更低（均P<0.05）；Andro以剂量依赖性的方式抑制Ishikawa/DPP细胞的增殖（均P<0.05）。Ishikawa/DPP细胞中Fas、FasL的表达水平均高于Ishikawa细胞（均P<0.05）。选取20 μg/mL DDP和25 μmol/L Andro为干预剂量，干预时间24 h。与对照组比较，DDP组Ishikawa/DPP细胞中PD-L1、MDR-1、Fas、FasL mRNA及蛋白表达水平显著升高（P<0.05），而克隆形成率、迁移与侵袭细胞数、凋亡率差异均无统计学意义（均P>0.05）；与DDP组比较，Andro组Ishikawa/DPP细胞中Fas、FasL mRNA表达水平、细胞克隆形成率、迁移与侵袭细胞数、PCNA、Bcl-2、MMP-2、PD-L1、MDR-1、Fas、FasL蛋白表达水平显著降低，BAX蛋白表达水平及凋亡率显著升高（P<0.05或P<0.01），pcDNA3.1-NC组与Andro组类似；与pcDNA3.1-NC组比较，pcDNA3.1-Fas/FasL组Ishikawa/DPP细胞上述指标变化均被逆转（P<0.05）。结论：Andro可能通过抑制Fas/FasL信号轴来抑制Ishikawa/DPP细胞增殖、迁移与侵袭，促进凋亡，从而降低细胞对DDP的耐药性。

Objective: To investigate the characteristics of newly reported HIV/AIDS cases aged 15 years and older in Shangcheng District, Hangzhou City from 2006 to 2022, so as to provide the basis for improving AIDS prevention and control strategies. Methods: Data of newly reported HIV/AIDS cases aged 15 years and older in Shangcheng District from 2006 to 2022 were collected through HIV/AIDS Comprehensive Control System of Chinese Disease Prevention and Control Information System. Population distribution and transmission routes were analyzed, and changing trends in case number were analyzed using average annual percent change (AAPC) and annual percent change (APC). Results: A total of 4 409 HIV/AIDS cases aged 15 years and older were newly reported in Shangcheng District from 2006 to 2022, including 3 932 males (89.18%). There were 3 447 cases (78.18%) under 50 years old and 962 cases (21.82%) being 50 years and older. Sexual contact was a predominant transmission route, with 4 326 cases accounting for 98.12%, including 2 626 cases (59.56%) with homosexual contact and 1 700 cases (38.56%) with heterosexual contact. The number of HIV/AIDS cases showed an overall upward trend from 2006 to 2022 (AAPC=13.038%, P<0.05), with an upward trend from 2006 to 2015 (APC=42.578%, P<0.05) and a downward trend from 2015 to 2022 (APC=-19.713%, P<0.05). The increase in the number of cases aged 50 years and older group was faster than that of cases aged under 50 years (AAPC=22.641% vs. 11.162%, both P<0.05). The increase in the number of cases with homosexual contact transmission was faster than that of cases with heterosexual contact transmission (AAPC=20.417% vs. 7.455%, both P<0.05). Conclusions The number of newly reported HIV/AIDS cases aged 15 years and older in Shangcheng District performed an overall upward trend from 2006 to 2022. The cases aged 50 years and older and transmitted through homosexual contact increased rapidly, which should be taken seriously.

DNA polymerase theta (Polθ), also known as DNA polymerase θ, is the member of the DNA polymerase A family and plays a crucial role in the repair of DNA double-strand breaks (DSB). Polθ has 3 distinct structural domains: the N-terminal helicase-like domain with a conserved sequence, the C-terminal polymerase domain, and the central domain, which is a disordered sequence connecting these two regions. Notably, Polθ is the only known polymerase in eukaryotes that possesses helicase activity. However, it is also an error-prone polymerase. When DNA DSBs occur, a specialized network consisting of at least 4 pathways, including classical-non homologous end joining (C-NHEJ), homologous recombination (HR), single-strand annealing (SSA), and alternative-end joining (Alt-EJ), is responsible for repairing DNA damage caused by DSBs. In the absence of major DNA repair pathways like HR, cells rely on Alt-EJ pathway mediated by Polθ to repair damaged DNA and maintain genomic stability. Nevertheless, due to the low fidelity of Polθ, Alt-EJ repair often leads to errors. Depletion of Polθ has shown to increases DSB formation and compromise genomic stability. Conversely, overexpression of Polθ has been associated with increases DNA damage markers and impairs cell cycle progression. As a result, the impact of Polθ on genome stability remains controversial. Furthermore, overexpression of Polθ is frequently observed in cancer and is associated with a characteristic mutational signature and poor prognosis. Depleting Polθ in an HR-deficient background has been shown to impair cell viability, suggesting a synthetic lethal (SL) relationship between Polθ and HR factors. In recent years, targeted chemotherapy drugs that inhibit tumor growth have gained significant attention. However, off-target effects and drug resistance pose challenges for clinical application, particularly with poly-ADP-ribose polymerase inhibitor (PARPi). Blocking Polθ activity in HR-deficient tumor cells has been found to reverse PARPi resistance, making Polθ a very promising therapeutic target in cancer treatment. The availability of crystal structures for both helicase and polymerase domain has facilitated the design of potent inhibitors of Polθ. Currently, several highly specific and effective small molecule inhibitors targeting Polθ, such as Novobiocin, RP-6685, and ART558, have been reported to effectively block various cancers with HR deficiency. The initial success of these inhibitors points to new directions for treating BRCA1/2-mutated tumors. Additionally, reducing the Alt-EJ repair pathway mediated by Polθ can improve HR repair efficiency and increase the chance of exogenous gene target integration (TI), suggesting potential new applications for Polθ inhibitors. This article reviews the recent research progress on the molecular function of Polθ and its involvement in the Alt-EJ pathway modification mechanism, providing insights for a deeper understanding of this field.

Objective:To explore the effect of family-based body weight management on cardiac function and readmission in patients with chronic heart failure.Methods:The study was a single-blind randomized controlled trial. Elderly patients with heart failure who were treated in Yichuan Community Health Service Center of Putuo District from June 2019 to May 2020 were enrolled in the study. The patients were randomly divided into the control group and the intervention group. All patients were treated with anti-heart failure drugs; in addition, the intervention group received family-based weight management and the control group received the conventional weight management. The cognition of self-management and weight management of patients was assessed by the Heart Failure Patient Self-Management Scale and the Weight Management Scale, before intervention and after 12 months of intervention, respectively. At the same time, the 6-min walking test, the New York Heart Association (NYHA) cardiac function grading assessment were performed, plasma N-terminal proB-type natriuretic peptide (NT-proBNP) was measured, left ventricular ejection fraction (LVEF) was determined and body weight measurement was completed; and the readmissions of patients due to heart failure during follow-up were recorded.Results:A total of 249 patients aged (65.2±2.9) years, including 104 males (41.8%) were enrolled; there were 124 in the intervention group and 125 in the control group. There were no significant differences in age, gender, marital status and educational level between the two groups (all P>0.05). There were no significant differences in the baseline scores of the Heart Failure Patient Self-Management Scale between the two groups (all P>0.05). After intervention, all scores of the intervention group were significantly better than those of the control group (all P<0.01). There were no significant differences in the scores of the Weight Management Scale between the two groups before intervention (all P>0.05), whereas all scores of the intervention group were significantly better than those of the control group after intervention (all P<0.01). There was no significant difference in body weight between the two groups before intervention ( P=0.397), while the average body weight of the intervention group was significantly lower than that of the control group after intervention ( P=0.029). At baseline, there were no significant differences in the LVEF, NT-proBNP level, 6-min walking distance, and the proportion of NYHA patients with grade Ⅲ/Ⅳ heart function between the two groups (all P>0.05). After intervention, LVEF and 6-min walking distance of patients in the intervention group were significantly higher than those in the control group (all P<0.01); the plasma NT-proBNP level and the proportion of NYHA grade Ⅲ/Ⅳ were significantly lower than those in the control group (all P<0.01). During follow-up, the rate of readmission due to heart failure in the intervention group was lower than that in the control group ( P<0.001). Conclusion:Family-based weight management can improve heart function and reduce the readmission rate in elderly patients with heart failure.

Objective:To examine the distribution features of angiotensin converting enzyme ( ACE) gene I/D polymorphisms and their correlation with left ventricular hypertrophy (LVH) in children and adolescents with essential hypertension. Methods:This was a case-control study.One hundred and forty-four children and adolescents who were diagnosed with essential hypertension but received no treatment at the Department of Cardiology, Children′s Hospital, Capital Institute of Pediatrics from January 2022 to April 2023 were recruited. ACE gene polymorphisms were detected using the polymerase chain reaction-restriction fragment length polymorphism method.Patients were divided into the DD+ DI genotype group and the II genotype group according to the dominant model, and the differences in gene frequencies, genotype frequencies, clinical data, and plasma renin-angiotensin-aldosterone system (RAAS) levels were compared between the two groups.In addition, according to the echocardiographic findings, patients were divided into the LVH and non-LVH groups.The risk factors of LVH were analyzed using the multivariate Logistic regression model.The correlation of ACE gene I/D polymorphisms with LVH in children and adolescents with essential hypertension was evaluated using the Spearman analysis.The correlation between plasma RAAS levels and LVH was assessed using the partial correlation analysis. Results:The frequencies of DD, DI and II genotypes in 144 patients were 11.1%, 50.7% and 38.2%, respectively.The frequency of the D and I allele was 36.5%(105/288) and 63.5%(183/288) respectively.Patients in the DD+ DI genotype group (89 cases) were older than those in the II genotype group (55 cases) ( Z=-2.308, P=0.021), and there was no statistically significant difference between the two groups in gender and height (all P>0.05).The incidence of LVH in the DD+ DI genotype group was higher than that in the II genotype group, with a statistically significant difference ( χ2=5.230, P=0.022).Renin activity in RAAS levels was positively correlated with left ventricular mass index and relative wall thickness ( r=0.276, 0.247; P=0.002, 0.006).Multivariate Logistic regression analysis showed that both DD+ DI genotype ( OR=5.678, 95% CI: 1.623-19.872) and body mass index (BMI) ( OR=1.124, 95% CI: 1.024-1.233) were risk factors for the development of LVH in children and adolescents with essential hypertension (all P<0.05). Conclusions:ACE gene I/D polymorphisms are strongly associated with LVH.DD+ DI genotype and BMI are independent risk factors for the development of LVH in children and adolescents with essential hypertension.

Objective To evaluate the protective effect and underlying mechanism of ulinastatin on hepatic ischemia-reperfusion injury.Methods Twenty-four male SD rats were divided into three groups:sham operation group(Sham group),hepatic ischemia-reperfusion injury group(HIRI group)and hepatic ischemia-reperfusion injury+ulinastatin pretreatment group(HIRI+UTI group),with 8 rats in each group.The HIRI rat models were established by occluding hepatic portal vein and hepatic artery for 1 h.In the HIRI+UTI group,the rats were intraperitoneally injected with ulinastatin at 30 min before model establishment,and an equivalent amount of normal saline was given in the Sham and HIRI groups.The rats were sacrificed at 6 h after model establishment.Serum samples were collected to detect alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels.Pathological changes of liver tissues were observed by hematoxylin-eosin(HE)staining.Ultrastructural changes of mitochondria in liver tissues were observed by transmission electron microscopy.The expression of glutathione peroxidase 4(GPX4)was determined by immunofluorescent staining.The contents of malondialdehyde(MDA),glutathione(GSH),Fe,reactive oxygen species(ROS)and GPX4 were detected.The expression levels of GPX4 and acyl-CoA synthetase long-chain family 4(ACSL4)messenger RNA(mRNA)and proteins in liver tissue were measured.Results Compared with the Sham group,serum ALT and AST levels were up-regulated,pathological changes such as congestion,hepatocyte necrosis and abnormal hepatic lobule structure were observed,pathological score was increased,the mitochondria shrank,the membrane density was increased,the mitochondrial crest was damaged or even absent,the contents of ROS,MDA and Fe were elevated,the GSH content was decreased,the fluorescent intensity of GPX4 was weakened,the relative expression levels of ACSL4 mRNA and protein were up-regulated,and the relative expression levels of GPX4 mRNA and protein were down-regulated in the HIRI group(all P＜0.05).Compared with the HIRI group,serum ALT and AST levels were down-regulated,liver tissue injury was alleviated,pathological score was decreased,mitochondrial shrinkage and crest breakage were mitigated,the contents of ROS,MDA and Fe were down-regulated,the GSH content was up-regulated,the fluorescent intensity of GPX4 was enhanced,the relative expression levels of ACSL4 mRNA and protein were down-regulated,and the relative expression levels of GPX4 mRNA and protein were up-regulated in the HIRI+UTI group(all P＜0.05).Conclusions Ulinastatin may alleviate hepatic ischemia-reperfusion injury in rats probably through inhibiting ferroptosis.