1.The Refinement and Innovation of The UV Cross-linking and Immunoprecipitation
Jia-Min ZHAO ; Cheng-Jiang LU ; Ming YANG ; Nashun BUHE ; Gang WANG
Progress in Biochemistry and Biophysics 2025;52(4):1036-1052
RNA-binding proteins (RBPs) are ubiquitous components within cells, fulfilling essential functions in a myriad of biological processes. These proteins interact with RNA molecules to regulate gene expression at various levels, including transcription, splicing, transport, localization, translation, and degradation. Understanding the intricate network of RBP-RNA interactions is crucial for deciphering the complex regulatory mechanisms that govern cellular function and organismal development. Ultravidet (UV) cross-linking and immunoprecipitation (CLIP) stands out as a powerful approach designed to map the precise locations where RBPs bind to RNA. By using UV light to create covalent bonds between proteins and RNA, followed by immunoprecipitation to isolate the protein-RNA complexes, researchers can identify the direct targets of specific RBPs. The advent of high-throughput sequencing technologies has revolutionized CLIP, enabling the identification of not only the types but also the exact sequences of RNA bound by RBPs on a genome-wide scale. The evolution of CLIP has led to the development of specialized variants, each with unique features that address specific challenges and expand the scope of what can be studied. High-throughput sequencing CLIP (HITS-CLIP) was one of the first advancements, significantly increasing the throughput and resolution of RNA-protein interaction mapping. Photoactivatable-ribonucleoside-enhanced CLIP (PAR-CLIP) introduced the use of photoactivatable ribonucleosides to enhance cross-linking efficiency and specificity, reducing background noise and improving the detection of low-abundance RNA-protein interactions. Individual-nucleotide resolution CLIP (iCLIP) further refined the technique, achieving unprecedented precision by resolving individual nucleotides involved in RBP binding, which is particularly valuable for studying the fine details of RNA structure and function. Despite the remarkable progress, there remains room for improvement in CLIP technology. Researchers continue to seek methods to increase sensitivity, reduce technical variability, and improve the reproducibility of results. Advances in sample preparation, data analysis algorithms, and computational tools are critical for addressing these challenges. Moreover, the application of CLIP to more diverse biological systems, including non-model organisms and clinical samples, requires the development of tailored protocols and the optimization of existing ones. Looking forward, the field of RNA biology is poised to benefit greatly from ongoing innovations in CLIP technology. The exploration of non-canonical RNA-protein interactions, such as those involving long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs), promises to reveal new layers of cellular regulation and may lead to the discovery of novel therapeutic targets. Furthermore, integrating CLIP data with other omics approaches, such as proteomics and metabolomics, will provide a more comprehensive understanding of the dynamic interplay between RNA and its binding partners within the cell. In conclusion, the continuous refinement and expansion of CLIP techniques have not only deepened our knowledge of RNA biology but have also opened up new avenues for investigating the molecular underpinnings of health and disease. As the technology matures, it is expected to play an increasingly pivotal role in both basic and applied research, contributing to the advancement of medical science and biotechnology.
2.The correlation between abnormal metabolic indexes and the severity of coronary artery lesions in patients with acute coronary syndrome
Yajun ZHAO ; Ming LIU ; Yuxiang DAI ; Xiaopan LI ; Xuelin CHENG ; Qizhe WANG ; Ru LIU ; Yaxin XU ; Sunfang JIANG
Chinese Journal of Clinical Medicine 2025;32(3):441-448
Objective To explore the influencing factors of coronary artery lesion severity in patients with acute coronary syndrome (ACS). Methods Clinical data of ACS patients admitted to Zhongshan Hospital, Fudan University from December 2017 to December 2019 were consecutively collected. The modified Gensini score was used to assess the severity of coronary artery lesions. Univariate and multivariate linear regression analyses were performed to identify independent factors associated with coronary artery lesion severity. Results A total of 1 689 ACS patients were included, with an average age of (64.04±11.45) years; 1 353 (80.11%) were male, and the mean modified Gensini score was (8.12±4.03). Multivariate linear regression analysis revealed that sex (β=0.97, P=0.001), age (β=0.03, P=0.021), estimated glomerular filtration rate (eGFR; β=-0.03, P<0.001), low-density lipoprotein cholesterol (LDL-C; β=0.58, P<0.001), apolipoprotein A1 (Apo A1; β=-1.28, P=0.012), lipoprotein(a) [Lp(a); β=0.001, P=0.033], and glycated hemoglobin A1C (HbA1C; β=0.45, P<0.001) were independent influencing factors of the modified Gensini score. Conclusions Metabolic indicators, including Apo A1, LDL-C, HbA1C, and Lp(a), may serve as risk factors for coronary artery lesion severity in ACS patients, with Apo A1 demonstrating the strongest impact.
3.Comparative analysis of the efficacy of RAPN and LPN in treating tumors in isolated kidney
Bin JIANG ; Yin LU ; Xupeng ZHAO ; Qiang CHENG ; Qing AI ; Fan GAO ; Hongzhao LI
Chinese Journal of Urology 2024;45(1):6-11
Objective:To compare the outcomes of robot-assisted laparoscopic partial nephrectomy (RAPN) and laparoscopic partial nephrectomy (LPN) in the treatment of tumors in isolated kidney, and analyze the factors influencing postoperative renal function and long-term survival in patients.Methods:A retrospective analysis was conducted on clinical data of 67 patients with tumors in isolated kidney who underwent surgery at the Chinese PLA General Hospital from November 2010 to January 2022. There were 48 males and 19 females, with an average age of (58.6±10.1) years old. The patients were divided into RAPN group (43 cases) and LPN group (24 cases) based on the surgical approach. The RAPN group had a higher R.E.N.A.L. score than the LPN group [(8.7±1.5) vs. (7.9±1.7), P=0.042]. There were no statistically significant differences between the two groups in terms of age [(57.4±10.2) years old vs. (60.9±9.8) years old, P=0.185], body mass index (BMI) [(25.7±3.5) kg/m 2 vs. (25.1±3.6) kg/m 2, P=0.518], and preoperative serum creatinine [(102.9±31.6) μmol/L vs. (102.3±22.4) μmol/L, P=0.930]. Twelve cases underwent hypothermic treatment during surgery, with 9 cases(20.9%) in the RAPN group and 3 cases(12.5%) in the LPN group( P=0.596). Surgical time, intraoperative warm ischemia time, intraoperative blood loss, postoperative fasting time, perioperative complication rate, postoperative serum creatinine, and other indicators were compared between the two groups. Multiple linear regression analysis was used to identify factors affecting postoperative serum creatinine. Kaplan-Meier curves were employed to analyze patient prognosis, and log-rank tests were performed to compare the differences between the two groups. Multiple Cox regression analysis was used to identify factors influencing patient prognosis. Results:All surgeries were completed successfully with negative pathological margins. There were no statistically significant differences between the RAPN and LPN groups in terms of surgical time [(136.6±47.6) min vs. (125.3±34.4) min, P=0.311], intraoperative ischemia time [23.0 (16.0, 30.0) min vs. 19.0 (13.5, 27.5) min, P =0.260], intraoperative blood loss [50.0 (50.0, 100.0) ml vs. 50.0 (22.5, 100.0) ml, P=0.247], postoperative hospital stay [(6.6±3.5) days vs. (7.7±4.2) days, P=0.244], time to drain removal [4(3, 5) days vs. 5(3, 6) days, P =0.175], postoperative fasting time [(2.1±0.7) days vs. (2.2±1.0) days, P=0.729], perioperative complication rate [18.6% (8/43) vs. 16.7% (4/24), P=1.000], postoperative serum creatinine [145.2 (128.3, 191.3) μmol/L vs. 157.8 (136.2, 196.3) μmol/L, P =0.229], and pathological staging [T 1a/T 1b/T 2a/T 3a/T 4 stage: 32/7/1/3/0 case vs. 17/5/0/1/1 case, P=0.804]. Kaplan-Meier survival curves showed that the total survival rates at 1, 3, and 5 years after surgery were 94.7%, 84.9%, and 84.9% for the RAPN group, and 100.0%, 95.5%, and 95.5% for the LPN group, with no statistically significant difference in the log-rank test ( P=0.116). Excluding 10 patients with preoperative tumor metastasis (7 in the RAPN group and 3 in the LPN group), the progression-free survival rates at 1, 3, and 5 years after surgery were 84.8%, 81.1%, and 81.1% for the RAPN group, and 100.0%, 95.0%, and 90.0% for the LPN group, with no statistically significant difference in the log-rank test ( P =0.142). Multiple linear regression analysis showed that the use of hypothermic treatment during surgery significantly reduced postoperative serum creatinine ( B=-72.191, P=0.048). Multiple Cox regression analysis revealed that BMI ( HR=0.743, P=0.044), pathological T stage ( HR=4.235, P=0.018), and preoperative metastasis ( HR=18.829, P=0.035) were independent factors affecting patient overall survival time. A smaller BMI, higher pathological stage, and preoperative metastasis were associated with poorer prognosis. Conclusions:Despite the higher R. E.N.A.L. score and greater surgical difficulty in the RAPN group, RAPN achieved similar perioperative and prognostic results as the LPN, indicating RAPN advantages in treating tumors in isolated kidney. Appropriate intraoperative hypothermic treatment can better protect postoperative renal function. BMI, pathological T stage, and preoperative metastasis are independent factors affecting overall survival time.
4.Study on The Promotion of Tenocyte Proliferation and Differentiation by Oriented Fiber Membrane Loaded With Nano-zinc Oxide
Jia FAN ; Peng-Cheng GU ; Xi-Ting CHENG ; Qiang JIANG ; Ya ZHAO ; Xiao-Fang PAN ; Yan BAI
Progress in Biochemistry and Biophysics 2024;51(8):1895-1903
ObjectiveTo simulate the microstructure and mechanical properties of tendon tissue and promote its regeneration and repair, electrospinning technology was used to prepare L-polylactic acid (PLLA) fiber membranes loaded with different nano zinc oxide contents and with oriented structures. Physical and chemical characterization and biological performance evaluation were carried out to explore their effects on tendon cell proliferation and differentiation. MethodsPreparation of PLLA fiber scaffolds and PLLA/ZnO fiber scaffolds containing different mass fractions of nano ZnO was performed using electrospinning technology. The physicochemical properties of the scaffold were characterized by scanning electron microscopy, mechanical stretching, and EDS spectroscopy. The scaffold was co-cultured with mouse tendon cells to detect its biocompatibility and regulatory effects on cell differentiation behavior. ResultsThe fiber scaffolds were arranged in an oriented manner, and zinc elements were uniformly distributed in the fibers. The tensile strength and Young’s modulus of PLLA/0.1%ZnO fiber scaffolds were significantly higher than PLLA fiber scaffolds. The number of cells on the surface of PLLA/0.1%ZnO fiber scaffold was significantly higher than that of the PLLA group, and the activity was better; mouse tendon cells exhibit directional adhesion and growth along the fiber arrangement direction. ConclusionThe oriented PLLA/0.1%ZnO fiber scaffold had excellent physicochemical properties, which can significantly promote the oriented growth, proliferation and differentiation of tendon cells. It is expected to be used for tendon tissue regeneration and repair in the future.
5.Analysis of the Institutional Distribution of Curative Care Expenditure for the Elderly Population before and after the Comprehensive Reform of Medical-pharmaceutical Separation and Linkage of Medical Con-sumption in Beijing
Yan JIANG ; Yiru ZHOU ; Xiaowei MAN ; Liying ZHAO ; Wei CHENG
Chinese Hospital Management 2024;44(1):31-35
Objective By studying the changes in the institutional distribution of curative care expenditure(CCE)of the elderly population before and after the comprehensive reform medical-pharmaceutical separation and linkage of medical consumption,it provided data reference for the next step of accurately optimizing the elderly patients flow.Methods A multi-stage stratified whole-group sampling survey was used to select the sample.A System of Health Accounts 2011 was used to calculate the CCE of elderly patients in medical institutions.Results The CCE of medical institutions for the elderly population in Beijing increased from 60.457 billion yuan to 797.54 billion yuan,with an average annual growth rate of 6.83%,the fastest growth rate of 24.04%for community-based health treat-ment center.The percentage of CCE in the community increased from 11.31%to 17.71%,while the percentage of CCE in tertiary hospitals decreased by 4.39 percentage points.The flow of CCE for outpatient patients was obviously opti-mized.Younger elderly outpatient patients are more willing to seek treatment in the community,but the flow di-rection of outpatient treatment for elderly patients is more optimized.The CCE fpr elderly outpatient patients with chronic diseases such as endocrine,nutritional and metabolic diseases and nervous system diseases have been substantially transferred to the community-based health center.Conclusion The reform has different impacts on the treatment of elderly patients with different genders,ages and diseases.It is necessary to strengthen the service capacity building of primary medical institutions,highlight the development characteristics of secondary hospitals,and accurately improve the hierarchical diagnosis and treatment system for elderly patients.
6.The Role and Research Progress of Communication in the Implementation of Hospice Care for Professional Health Caregivers
Jingjing ZHAO ; Enshe JIANG ; Lexin WANG ; Fang CHENG
Chinese Medical Ethics 2024;35(6):663-668
The communication between professional health caregivers, which mainly composed of medical staff, and patients and their families plays an important role in hospice care. The effective doctor-patient communication can guide terminal patients and their families to establish a correct medical view, reduce excessive medical treatment, and improve the quality of life for terminal patients. This paper briefly described the communication context of hospice care, the communication subject and the current situation of hospice care, analyzed the problems and challenges encountered by professional health caregivers, which mainly including medical staff, in the communication context of hospice care diagnosis and hospice care, and made the prospects of the development of effective communication measures.
7.Methodology for the Development of Clinical Practice Guidelines for Chinese Patent Medicine(Part 3): Identification of Clinical Questions
Ziteng HU ; Ning LIANG ; Lijiao YAN ; Yujing ZHANG ; Yaxin CHEN ; Fuqiang ZHANG ; Zhao CHEN ; Yin JIANG ; Cuicui CHENG ; Nannan SHI ; Yanping WANG
Journal of Traditional Chinese Medicine 2024;65(1):55-59
The identification of clinical questions for clinical practice guidelines of Chinese patent medicine (CPM) is important for subsequent evidence retrieval, evaluation of evidence quality, formation of recommendations. This paper described a methodological proposal for the identification of clinical questions for CPM guidelines to highlight the characteristics of Chinese patent medicine and reflect its effect in specific stage of the disease. Considering four aspects, namely, the drug of Chinese patent medicine (D), the specific disease stage (S), comparison (C), and specific outcome (O), DSCO framework has been proposed to formulate the clinical questions. Multi-source information through scientific research, policy or standard documents, and clinical data are suggested for collecting clinical questions, and clear selection criteria should be set to finalize the clinical questions to be addressed by the guideline. In addition, the above process needs to be transparently and publicly reported in order to ensure the clarity and completeness of the guidelines.
8.Optimization of water extraction technology of Xiangqin jiere granules by orthogonal design based on G1-entropy weight compared with BP neural network
Bingduo CHENG ; Liqin LUO ; Yuanzeng LI ; Jie JIANG ; Yiying CHEN ; Ji ZHAO ; Rui XUE ; Yunshu MA
China Pharmacy 2024;35(1):27-32
OBJECTIVE Optimizing the water extraction technology of Xiangqin jiere granules. METHODS The orthogonal test of 3 factors and 3 levels was designed, and comprehensive scoring was conducted for the above indexes by using G1-entropy weight to obtain the optimized water extraction technology of Xiangqin jiere granules with water addition ratio, extraction time and extraction times as factors, using the contents of forsythoside A, baicalin, phillyrin, oroxylin A-7-O-β-D-glycoside, wogonoside, baicalein and wogonin, and extraction rate as evaluation indexes. BP neural network modeling was used to optimize the network model and water extraction process using the results of 9 groups of orthogonal tests as test and training data, the water addition multiple, decocting time and extraction times as input nodes, and the comprehensive score as output nodes. Then the two analysis methods were compared by verification test to find the best water extraction process parameters. RESULTS The water extraction technology optimized by the orthogonal test was 8-fold water, extracting 3 times, extracting for 1 h each time. Comprehensive score was 96.84 (RSD=0.90%). The optimal water extraction technology obtained by BP neural network modeling included 12-fold water, extracting 4 times, extracting for 0.5 h each time. The comprehensive score was 92.72 (RSD=0.77%), which was slightly lower than that of the orthogonal test. CONCLUSIONS The water extraction technology of Xiangqin jiere granules is optimized successfully in the study, which includes adding 8-fold water, extracting 3 times, and extracting for 1 hour each time.
9.Multicenter evaluation of the diagnostic efficacy of jaundice color card for neonatal hyperbilirubinemia
Guochang XUE ; Huali ZHANG ; Xuexing DING ; Fu XIONG ; Yanhong LIU ; Hui PENG ; Changlin WANG ; Yi ZHAO ; Huili YAN ; Mingxing REN ; Chaoying MA ; Hanming LU ; Yanli LI ; Ruifeng MENG ; Lingjun XIE ; Na CHEN ; Xiufang CHENG ; Jiaojiao WANG ; Xiaohong XIN ; Ruifen WANG ; Qi JIANG ; Yong ZHANG ; Guijuan LIANG ; Yuanzheng LI ; Jianing KANG ; Huimin ZHANG ; Yinying ZHANG ; Yuan YUAN ; Yawen LI ; Yinglin SU ; Junping LIU ; Shengjie DUAN ; Qingsheng LIU ; Jing WEI
Chinese Journal of Pediatrics 2024;62(6):535-541
Objective:To evaluate the diagnostic efficacy and practicality of the Jaundice color card (JCard) as a screening tool for neonatal jaundice.Methods:Following the standards for reporting of diagnostic accuracy studies (STARD) statement, a multicenter prospective study was conducted in 9 hospitals in China from October 2019 to September 2021. A total of 845 newborns who were admitted to the hospital or outpatient department for liver function testing due to their own diseases. The inclusion criteria were a gestational age of ≥35 weeks, a birth weight of ≥2 000 g, and an age of ≤28 days. The neonate′s parents used the JCard to measure jaundice at the neonate′s cheek. Within 2 hours of the JCard measurement, transcutaneous bilirubin (TcB) was measured with a JH20-1B device and total serum bilirubin (TSB) was detected. The Pearson′s correlation analysis, Bland-Altman plots and the receiver operating characteristic (ROC) curve were used for statistic analysis.Results:Out of the 854 newborns, 445 were male and 409 were female; 46 were born at 35-36 weeks of gestational age and 808 were born at ≥37 weeks of gestational age. Additionally, 432 cases were aged 0-3 days, 236 cases were aged 4-7 days, and 186 cases were aged 8-28 days. The TSB level was (227.4±89.6) μmol/L, with a range of 23.7-717.0 μmol/L. The JCard level was (221.4±77.0) μmol/L and the TcB level was (252.5±76.0) μmol/L. Both the JCard and TcB values showed good correlation ( r=0.77 and 0.80, respectively) and agreements (96.0% (820/854) and 95.2% (813/854) of samples fell within the 95% limits of agreement, respectively) with TSB. The JCard value of 12 had a sensitivity of 0.93 and specificity of 0.75 for identifying a TSB ≥205.2?μmol/L, and a sensitivity of 1.00 and specificity of 0.35 for identifying a TSB ≥342.0?μmol/L. The TcB value of 205.2?μmol/L had a sensitivity of 0.97 and specificity of 0.60 for identifying TSB levels of 205.2 μmol/L, and a sensitivity of 1.00 and specificity of 0.26 for identifying TSB levels of 342.0 μmol/L. The areas under the ROC curve (AUC) of JCard for identifying TSB levels of 153.9, 205.2, 256.5, and 342.0 μmol/L were 0.96, 0.92, 0.83, and 0.83, respectively. The AUC of TcB were 0.94, 0.91, 0.86, and 0.87, respectively. There were both no significant differences between the AUC of JCard and TcB in identifying TSB levels of 153.9 and 205.2 μmol/L (both P>0.05). However, the AUC of JCard were both lower than those of TcB in identifying TSB levels of 256.5 and 342.0 μmol/L (both P<0.05). Conclusions:JCard can be used to classify different levels of bilirubin, but its diagnostic efficacy decreases with increasing bilirubin levels. When TSB level are ≤205.2 μmol/L, its diagnostic efficacy is equivalent to that of the JH20-1B. To prevent the misdiagnosis of severe jaundice, it is recommended that parents use a low JCard score, such as 12, to identify severe hyperbilirubinemia (TSB ≥342.0 μmol/L).
10.Risk factors and prediction model of acute respiratory failure in patients with hypertriglyceridemic acute pancreatitis
Yaobing LIANG ; Zhenhua FU ; Ziyue ZHAO ; Jianming LUO ; Dongyu CHENG ; Haixing JIANG ; Shanyu QIN
Tianjin Medical Journal 2024;52(11):1183-1187
Objective To analyze risk factors of acute respiratory failure(ARF)in patients with hypertriglyceridemia acute pancreatitis(HTG-AP)and construct a risk prediction model.Methods A total of 222 HTG-AP patients were included in this study and divided into the non-ARF group(176 cases)and the ARF group(46 cases)according to diagnostic guidelines for ARF.Clinical data of the two groups were compared and the predictive factors were screened.These selected factors were then utilized in a multivariate Logistic regression analysis to construct a Logistic regression model.Subsequent evaluation of the model′s predictive ability,accuracy and clinical utility was conducted through ROC,curve analysis,calibration plot examination and decision curve analysis(DCA),respectively.Results Compared with the non-ARF group,the levels of high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL)-C and albumin(ALB)were decreased in the ARF group(P<0.05),while the levels of creatinine(Cr),urea nitrogen(BUN),aspartate aminotransferase(AST)and C-reactive protein(CRP)were increased,and the incidence of pleural fluid and ascites was also increased(P<0.05).Multivariate Logistic regression analysis showed that higher levels of Cr and AST,lower levels of ALB,HDL-C and ascites were independent risk factors for HTG-AP complicated ARF(P<0.05).Based on these results,a column-line prediction model for HTG-AP complicated ARF was established.After internal verification,the area under curve(AUC)of receiver operating characteristic(ROC)curve of the nomogram model was 0.952(95%CI:0.923-0.981),the Youden index was 0.808 and the sensitivity and specificity were 93.33%and 87.43%,respectively.The calibration curve showed that the probability of HTG-AP concurrent ARF predicted by the model was in good agreement with the actual probability.The DCA curve showed that the model had certain clinical value.Conclusion The nomogram prediction model combined could provide a scheme for the clinical prevention of HTG-AP complicated with ARF.

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