1.Expression and Clinical Significance of PLCβ4 Gene in Hepatocellular Carcinoma Analyzed Based on TCGA Database and Experimental Validation
Limei WEN ; Yali GUO ; Qiang HOU ; Dongxuan ZHENG ; Wu DAI ; Xiang GAO ; Jianhua YANG ; Junping HU
Cancer Research on Prevention and Treatment 2025;52(6):502-510
Objective To analyze the PLCβ4 gene mRNA expression and its clinical significance in hepatocellular carcinoma (HCC) based on TCGA database. Methods Based on the data on 424 clinical samples (including 374 cases of HCC tissues and 50 cases of nontumor liver tissues) in the TCGA database, Kaplan–Meier method, Cox regression analysis, and immune infiltration analysis were performed to evaluate the relationship between PLCβ4 gene and the clinical characteristics and survival prognosis of HCC patients. Correlation analysis between PLCβ4 gene and 24 types of immune cells was applied to investigate the relationship between PLCβ4 gene and immune cell infiltration and mRNA expression level of TP53 gene, a high-frequency mutation gene in HCC. In addition, paraffin sections of highly, moderately, and poorly differentiated tumor tissues and normal liver tissues from HCC patients were collected. The histopathological observation was carried out via HE staining method, and the expression levels of PLCβ4 and Ki-67 proteins in each clinical sample were verified through the immunohistochemical method. Results The expression level of PLCβ4 gene in HCC was significantly higher than that in normal tissues (P<0.01), and all patients in the PLCβ4 high-expression group had a significantly longer overall survival than those in the low-expression group (P<0.05), which suggested that PLCβ4 substantially affected the prognosis of HCC patients. Correlation analysis showed that the expression level of PLCβ4 gene was highly correlated with immune cell infiltration and the expression level of TP53 gene. As verified by clinical sample experiments, HE staining experiments and immunohistochemical results revealed that PLCβ4 gene expression in HCC tissue samples was significantly higher than that in normal tissues (P<0.001), and it was negatively correlated with the degree of differentiation. Conclusion PLCβ4 may serve as an independent prognostic factor in HCC and is expected to be a novel molecular target for HCC treatment.
2.An analysis of 6-month follow-up efficacy of low-intensity extracorporeal shock wave therapy for refractory prostate-pelvic syndrome
Xiao XU ; Guizhong LI ; Xiang DAI ; Jie WANG
Journal of Modern Urology 2025;30(2):157-160
Objective: To evaluate the long-term efficacy and safety of low-intensity extracorporeal shock wave therapy (Li-ESWT) for refractory prostate-pelvic syndrome (PPS). Methods: Clinical data of 173 patients with refractory PPS undergoing Li-ESWT at our hospital during Jan.2020 and Jan.2023 were retrospectively analyzed.All patients received weekly treatment for 8 consecutive weeks.Changes in the National Institutes of Health chronic prostatitis symptom index (NIH-CPSI),international prostate symptom score (IPSS),visual analog scale (VAS),and international index of erectile function-5 (IIEF-5) were compared before treatment,immediately,1,3,and 6 months after treatment. Results: A total of 142 patients (82.1%) completed all follow-ups.Compared to baseline data,there was a statistically significant improvement in NIH-CPSI,IPSS,VAS,and IIEF-5 scores immediately after treatment and 1,3,and 6 months after treatment (P<0.01).No significant adverse reactions or complications were observed throughout the follow-up.At the time of treatment completion,115 patients (81.0%) had a decrease of ≥6 in NIH-CPSI; 99 patients (69.7%) had a decrease of ≥3 in IPSS; 121 patients (85.2%) had a decrease of ≥3 in VAS; 105 patients (73.9%) had an increase of ≥4 in IIEF-5.At the 6-month follow-up,patients who responded to treatment maintained satisfactory therapeutic effects. Conclusion: Li-ESWT can significantly improve clinical symptoms and quality of life for patients with refractory PPS,with therapeutic effects lasting at least 6 months.
3.Effects of scutellarin on endometrial carcinoma Ishikawa cells based on PI3K/Akt signaling pathway
Li-Li DAI ; Jing WANG ; Xu-Rui WEI ; Qing-Ya MA ; Na WANG ; Zhi-Xiang DU
The Chinese Journal of Clinical Pharmacology 2024;40(1):27-31
Objective To study the effects of scutellarin on endometrial carcinoma Ishikawa cells,and analyze the correlation between the effects and phosphatidyl inositol 3 kinase(PI3 K)/protein kinase B(Akt)signaling pathway.Methods Ishikawa cells were divided into blank group and experimental-L,-M,-H groups,each group was treated with complete medium containing 0,5,10 and 20 μmol·L-1 scutellarin,respectively.Cell viability,clonal formation ability,metastatic ability,invasion,apoptosis and protein expression were detected by cell counting kit-8(CCK-8),plate cloning,scratch,Transwell,flow cytometry and Western blot assay,respectively.Results The cell viability of blank group and experimental-L,-M,-H groups at 48 h were(100.00±0.00)%,(78.51±7.54)%,(52.93±4.91)%and(41.62±5.33)%;the clone formation rate were(100.00±0.00)%,(56.59±6.34)%,(35.23±4.62)%and(10.66±1.91)%;the scratch healing rate were(53.70±6.19)%,(40.59±4.75)%,(34.25±4.40)%and(15.78±2.14)%;the number of invasive cells were 189.70±14.06,106.82±12.67,84.37±8.13 and 53.74±6.78;the relative expression levels of matrixmetallo proteinase-2 were 0.96±0.10,0.73±0.06,0.68±0.08 and 0.42±0.05;tissue inhibitor of MMP-1(TIMP-1)were 0.35±0.04,0.51±0.05,0.74±0.08 and 1.20±0.14;the apoptosis rates were(4.21±0.53)%,(15.83±2.42)%,(22.72±3.85)%and(34.41±4.67)%;the relative expression levels of B cell lymphoma-2(Bcl-2)were 1.38±0.15,0.90±0.10,0.56±0.06 and 0.24±0.03;Bcl-2 associated X protein(Bax)were 0.31±0.02,0.44±0.04,0.93±0.11 and 1.26±0.14;the relative expression levels of PI3Kp85 were 0.67±0.05,0.42±0.04,0.36±0.02 and 0.28±0.03;phosphorylated Akt(p-Akt)were 0.78±0.06,0.53±0.04,0.46±0.05 and 0.42±0.03.Compared with the blank group,the above indexes in the experimental-L,-M,-H groups were statistically significant(P<0.05 or P<0.01).Conclusion Scutellarin can inhibit the proliferation,metastatic ability and invasion of endometrial carcinoma Ishikawa cells and promote apoptosis by regulating PI3K/Akt signaling pathway.
4.Advances in HIV-1 latency-regulating agents
Jiao-jiao DAI ; Xiang-yi JIANG ; Da FENG ; Hao LIN ; Xin-yong LIU ; Peng ZHAN
Acta Pharmaceutica Sinica 2024;59(4):840-852
At present, there is no cure for acquired immune deficiency syndrome (AIDS) due to HIV-1 latent reservoirs. Therefore, it urgently requires novel HIV-1 latency-regulating agents with high potency, low toxicity and favorable drug-like properties to achieve a functional cure for AIDS. Herein, we reviewed the advances in HIV-1 latency-regulating agents since 2019, including the drug discovery strategies, bioactivities, and mechanisms of these compounds. It is of great guiding significance in the development of latency-regulating agents with clinical value.
5.Retrospective analysis of the epidemic characteristics of Gram-negative bacteria in a second-class hospital in Shanghai
Feifei DAI ; Xiang FU ; Qiongnian CHEN ; Suchun YU
Journal of Pharmaceutical Practice and Service 2024;42(12):528-532
Objective To retrospectively analyze the distribution and drug resistant characteristics of isolated pathogens (especially Gram-negative bacteria) in hospital and promote the rational use of antibacterial drugs. Methods The pathogens and drug sensitivity test results were collected from the whole hospital every quarter from July 2018 to March 2023, and were conducted retrospective analysis. Results A total of 14 844 pathogens were isolated and detected, including 4 757 Gram-positive bacteria (32.05%) and 10 087 Gram-negative bacteria (67.95%). The detection rate of extended spectrum beta-lactamases Escherichia coli was 43.23%, and it maintained high sensitivity to carbapenems and enzyme inhibitors, and the sensitivity rate was about 99.00%. The drug resistance of Klebsiella pneumoniae was more serious, and carbapenem-resistant, multi-drug resistance and extensively-drug resistance appeared. In 2022,the detection rate of carbapenem-resistant Klebsiella pneumoniae was 54.82%; the detection rates of carbapenem-resistant Pseudomonas Aeruginosa and carbapenem-resistant Acinetobacter baumannii in non-fermentative bacteria were 22.02% and 74.34%, the detection rate of multi-drug resistant Acinetobacter baumannii was >70%. Conclusion The problem of drug resistance of Gram-negative bacteria was still serious, especially in Klebsiella pneumoniae and Acinetobacter baumannii. The problem of bacterial resistance poses great challenges to clinical treatment, and it is necessary to conduct regular monitoring and promote rational drug use.
6.Evaluation of chemiluminescence immunoassay kit for detection of hepatitis D virus IgG antibody
Rongchen YUAN ; Fangming CHENG ; Kuanhui XIANG ; Yongcong LI ; Tianxun HUANG ; Zhenchao TIAN ; Xiongwei LIU ; Xiaozhong WANG ; Zhuanguo WANG ; Yahong MA ; Jing ZHOU ; Erhei DAI ; Chungen QIAN ; Tong LI ; Tao SHEN ; Bangning CHENG
Chinese Journal of Laboratory Medicine 2024;47(3):234-238
Objective:This study evaluates the performance of chemiluminescence assay, which is designed to detect Hepatitis D Virus (HDV) Immunoglobulin G (IgG) antibodies.Methods:A comparative analysis was conducted among chemiluminescence anti-HDV IgG reagent, the magnetic particle-based domestic reagent A and domestic reagent B, and the Robo Gene HDV RNA kit, using 1909 HBsAg-positive plasma samples. This comparison aimed to delineate clinical specificity and detection accuracy. The anti-HDV IgG reagent precision was assessed at three different concentration levels following the Clinical Laboratory Standards Institute EP5-A2 guidelines. The specificity of the assay was validated using 200 HAV IgM positive, 545 HBsAg-positive but anti-HDV IgG-negative, 350 anti HCV positive plasma samples and 200 healthy human blood samples. Additionally, a concordance study was conducted with 545 HBsAg-positive and 37 anti-HDV IgG-positive plasma samples, comparing the anti-HDV IgG reagent against reagent A.Results:1 909 HBsAg-positive plasma samples were tested using 3 anti HDV IgG reagent and 1 HDV RNA reagent, 19 samples were identified as anti-HDV IgG-positive. The anti-HDV IgG demonstrated superior accuracy and specificity. The assay exhibited excellent precision, with intra-assay coefficient of variation (CV) values ranging from 1.57% to 4.30%, and inter-assay CV values between 1.71% and 4.67% for detecting samples at high, medium, and low concentration levels. Concordance with Reagent A showed consistent results in both positive and negative detections.Conclusion:In this study, the anti-HDV IgG reagent (chemiluminescence method) displayed outstanding specificity in detecting clinical samples and exhibited a high conformity rate with commercialized reagents, making it potentially suitable for screening anti-HDV IgG in HBsAg-positive samples.
7.Assessment and preliminary clinical application of a domestic nucleic acid detection reagent for hepatitis D virus
Yongcong LI ; Rongchen YUAN ; Kuanhui XIANG ; Guomin OU ; Tianxun HUANG ; Fangming CHENG ; Zhenchao TIAN ; Xiongwei LIU ; Xiaozhong WANG ; Feng GUO ; Yahong MA ; Jing ZHOU ; Erhei DAI ; Bangning CHENG ; Tong LI ; Tao SHEN ; Chungen QIAN
Chinese Journal of Laboratory Medicine 2024;47(3):239-244
Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.
8.Advances in therapeutic drug monitoring methods based on liquid chromatography-tandem mass spectrometry
Ziying LI ; Jie XIE ; Ziyu QU ; You JIANG ; Di ZHANG ; Songlin YU ; Xiaoli MA ; Ling QIU ; Xinhua DAI ; Xiang FANG ; Xiaoping YU
Chinese Journal of Laboratory Medicine 2024;47(3):332-340
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology has the characteristics of high specificity and high throughput, making it rapidly applied and developed in the field of clinical testing. Its application in the monitoring of therapeutic drugs can effectively improve the quantitative accuracy and sensitivity, and formulate a personalized and optimal dosing plan for patients. However, this technology still faces some challenges, and automation, quality control, and quantitative traceability will be the future development direction.
9.Upregulation of α1-acidglycoprotein to ameliorate hepatocyte lipid accumulation by Astragaloside derivative HHQ16
Xiang LI ; Xianmin DAI ; Xia LIU ; Yang SUN
Journal of Pharmaceutical Practice and Service 2024;42(4):141-146
Objective To investigate the effect and mechanism of HHQ 16, a derivative of astragaloside Ⅳ, on hepatocyte lipid accumulation. Methods Free fatty acids were used to stimulate lipid hepatocyte accumulation. Triglyceride and Oil Red O staining were detected to reflect hepatocyte lipid accumulation. The expression of α1-acidglycoprotein (ORM) and its regulators were detected by real-time quantitative PCR and immunoblotting. The expression of ORM1 was interfered with siRNA to determine whether it mediated the action of HHQ16.The expression of ORM1 was interfered by siRNA to determine whether it mediated the action of HHQ16. Results HHQ16 significantly ameliorated FFA-induced hepatocyte lipid deposition. HHQ16 elevated ORM expression, and the protective effect of HHQ16 on hepatocyte lipid accumulation was reversed by ORM interference. Conclusion HHQ16 could ameliorate hepatocyte lipid accumulation by elevating ORM.
10.Surgical treatment decision and clinical value of molecularly defined renal carcinoma with highly aggressive behavior
Xiang LI ; Lijing XU ; Guangxi SUN ; Jindong DAI
Journal of Modern Urology 2024;29(4):293-297
Molecularly defined renal carcinoma is a pathologic subtype of renal cell cancer (RCC) with a definite driver gene, which was first proposed in the 2022 WHO classification of tumors of the urinary system and male reproductive organs. The fumarate hydratase-deficient renal cell carcinoma and SMARCB1-deficient renal medullary carcinoma are highly aggressive and lethal subtypes. Due to the low incidence and lack of research on the mechanism, there is almost no effective treatment for these aggressive RCC subtypes. Cytoreductive nephrectomy or metastasectomy are important methods to improve the survival and quality of life of metastatic RCC patients under effective systemic therapy. However, for the highly aggressive RCC, the clinical value of the above surgical strategies is still unclear. In this review, we will discuss these problems in order to provide reference for the improvement of prognosis for patients with highly aggressive RCC subtypes.

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