1. Mechanism of ellagic acid improving cognitive dysfunction in APP/PS double transgenic mice based on PI3K/AKT/GSK-3β signaling pathway
Li-Li ZHONG ; Xin LU ; Ying YU ; Qin-Yan ZHAO ; Jing ZHANG ; Tong-Hui LIU ; Xue-Yan NI ; Li-Li ZHONG ; Yan-Ling CHE ; Dan WU ; Hong LIU
Chinese Pharmacological Bulletin 2024;40(1):90-98
Aim To investigate the effect of ellagic acid (EA) on cognitive function in APP/PS 1 double- transgenic mice, and to explore the regulatory mechanism of ellagic acid on the level of oxidative stress in the hippocampus of double-transgenic mice based on the phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase-3 (PI3K/AKT/GSK-3 β) signaling pathway. Methods Thirty-two SPF-grade 6-month-old APP/PS 1 double transgenic mice were randomly divided into four groups, namely, APP/PS 1 group, APP/PS1 + EA group, APP/PS1 + LY294002 group, APP/PS 1 + EA + LY294002 group, with eight mice in each group, and eight SPF-grade C57BL/6J wild type mice ( Wild type) were selected as the blank control group. The APP/PS 1 + EA group was given 50 mg · kg
2.Colorimetric Determination of Antioxidant Capacity by Peroxidase Mimics Based on Ruthenium Nanoparticles Supported on Carbon Nanosheets
Ru-Xue HE ; Peng XU ; Fang-Ning LIU ; Peng-Juan NI ; Yi-Zhong LU
Chinese Journal of Analytical Chemistry 2024;52(1):45-53,中插5-中插13
Lattice strain ruthenium nanoparticles uniformly and stably supported on nitrogen-modified carbon nanosheets(RuNPs/NC)were prepared via simple wet-chemical and subsequent pyrolysis method.The nitrogen doped NC could effectively improve their uniform dispersion and lattice compression of RuNPs.Through changing the pyrolysis temperature,the nitrogen content,types and degree of lattice strain of RuNPs could be effectively tuned,which could be used to adjust and control their peroxidase-like activity.The as-prepared RuNPs/NC-900 exhibited highest peroxidase-like activity,and could catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine(TMB)to produce a blue product with the maximum absorption at 652 nm in the presence of H2O2.The steady-state kinetic analysis indicated that the reaction catalyzed by RuNPs/NC followed the Michaelis-Menten kinetic model.Tannic acid(TA),gallic acid(GA)and ascorbic acid(AA)could effectively inhibit the RuNPs/NC-H2O2-triggered chromogenic reaction of TMB,resulting in color fading and decrease in absorbance.Based on this,a sensitive and accurate system was constructed for detection of TA,GA and AA.The detection limits(3σ/S)for TA,GA and AA were 0.014,0.014 and 0.29 μmol/L,respectively.This study not only developed a colorimetric sensing method based on RuNPs/NC nanozyme but also offered a new approach for the sensitive detection of antioxidants in food.
3.Study on the Distribution Pattern of Traditional Chinese Medicine Syndromes in Patients with Dry Eye and Its Correlation with Gender and Age
Yu-Xuan LI ; Ni TIAN ; Lan YU ; Rui-Ying ZHONG
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(3):550-554
Objective To explore the etiology and pathogenesis of dry eye by studying the distribution pattern of gender,age and traditional Chinese medicine(TCM)syndrome type in dry eye patients and by analyzing their correlation.Methods A total of 244 patients with dry eye who met the inclusion criteria were selected.The distribution of gender,age and TCM syndrome types was statistically analyzed,and then the correlation of TCM syndrome types with gender and age of dry eye patients was explored.Results(1)Of the 244 dry eye patients,96(39.34%)were male and 148(60.66%)were female,the incidence of the female being higher than that of the male.There were 124(50.82%)patients younger than 45 years old,81(33.20%)patients aged 45-60 years old,and 39(15.98%)patients older than 60 years old.The proportion of the patients younger than 45 years old was higher than that of other age groups.(2)Among the 244 patients with dry eyes,89 cases(36.47%)were differentiated as liver and kidney deficiency syndrome,75 cases(30.74%)were differentiated as qi stagnation and blood stasis syndrome,69 cases(28.28%)were differentiated as spleen and kidney deficiency,and 11 cases(4.51%)were differentiated as yin deficiency and damp-heat syndrome.And the occurrence frequency of the above four syndrome types was in descending order.(3)In the dry eye patients of various age groups,patients aged<45 years old predominantly suffered from qistagnation and blood stasis syndrome,accounting for 41.94%(52/124);patients aged 45-60 years old and those aged>60 years old predominantly suffered from liver and kidney deficiency syndrome,accounting for 46.91%(38/81)and 53.85%(21/39),respectively.The distribution of TCM syndrome types varied in the patients with different age groups,and the difference was statistically significant(χ2 = 22.128,P<0.01).(4)In male dry eye patients,qi stagnation and blood stasis syndrome was predominant,accounting for 39.58%(38/96);among female dry eye patients,liver and kidney deficiency syndrome and spleen and kidney deficiency syndrome were prevalent,accounting for 41.89%(62/148)and 31.08%(46/148),respectively.The distribution of TCM syndrome types varied in the patients with different genders,and the difference was statistically significant(χ2 = 82.610,P<0.01).Conclusion The TCM syndromes of patients with dry eyes are frequently differentiated as liver and kidney deficiency syndrome,followed by the qi stagnation and blood stasis syndrome.The prevalence of dry eyes is related to the gender and age,and gender and age are correlated with the TCM syndrome types to certain extent.
4.The nomogram based on preoperative inflammatory biomarkers used for predicting the prognosis of HCC patients treated with transcatheter arterial chemoembolization:its construction and validation
Dongxu ZHAO ; Binyan ZHONG ; Zhongheng HOU ; Yi ZHAN ; Caifang NI
Journal of Interventional Radiology 2024;33(3):245-258
Objective To construct and validate a predictive model based on preoperative inflammatory biomarkers,and to evaluate its ability in predicting the prognosis of patients with unresectable hepatocellular carcinoma(HCC)after receiving transcatheter arterial chemoembolization(TACE).Methods A total of 544 patients with HCC,who received TACE as the initial treatment at six medical institutions between January 2007 and December 2020,were retrospectively collected.The patients were divided into training cohort(n=376)and validation cohort(n=168).LASSO algorithm and Cox regression analysis were used to screen out the independent influencing factors and to make modelling.The model was validated based on the discrimination,calibration and clinical applicability,and the Kaplan-Meier risk stratification curves were plotted to determine the prognostic differences between groups.The likelihood ratio chi-square value,R2 value,akaike information criterion(AIC)value,C-index and AUROC value of the model were calculated to determine its accuracy and efficiency.Results The training cohort and validation cohort had 376 participants and 168 participants respectively.Multivariate analysis indicated that BCLC,tumor size,number of tumor lesions,neutrophil and prognostic nutritional index(PNI)were the independent influencing factors for postoperative overall survival(OS),with all P being<0.05;the BCLC grade,tumor size,number of tumor lesions,NLR,PNI and PS score were the independent influencing factors for progression-free survival(PFS),with all P being<0.05.The C-indexes of the OS and PFS models were 0.735(95% CI=0.708-0.762)and 0.736(95% CI=0.711-0.761)respectively,and the external validation was 0.721(95% CI=0.680-0.762)and 0.693(95% CI=0.656-0.730)respectively.Ideal discrimination ability of the nomogram was exhibited in time-dependent C-index,time-dependent ROC,and time-dependent AUC.The calibration curves significantly coincided with the ideal standard lines,indicating that the model had high stability and low over-fitting level.Decision curve analysis revealed that there was a wider range of threshold probabilities and it could augment net benefits.The Kaplan-Meier curves for risk stratification indicated that the prognosis of patients varied dramatically between risk categories(P<0.000 1).The Kaplan-Meier curves for risk stratification indicated that the prognosis of patients varied dramatically among different risk groups(P<0.000 1).The likelihood ratio chi-square value,R2 value,AIC value,C-index and AUROC value of the model were better than those of other models commonly used in clinical practice.Conclusion The newly-developed prognostic nomogram based on preoperative inflammatory indicators has excellent accuracy as well as excellent prediction effect in predicting the prognosis of patients with unresectable HCC after receiving TACE,therefore,it can be used as an effective tool for guiding individualized treatment and for predicting prognosis.(J Intervent Radiol,2024,33:245-258)
5.Toxicity and Mechanism of Di-(2-ethylhexyl) Phthalate on Testis
An-Ni FENG ; Ren-Ren SUN ; Yu-Bo XIAO ; Zhao-Ming ZENG ; Zhong-Cheng MO ; Yuan-Jie XIE
Progress in Biochemistry and Biophysics 2024;51(3):555-563
Di-(2-ethylhexyl) phthalate (DEHP) is currently one of the most widely used plasticizers, widely found in all kinds of items, such as children’s toys and food packaging materials, but also added to wallpaper, cable protective agents and other building decoration materials. DEHP is toxic and absorbed by the human body through respiratory tract, digestive tract and skin contact, which can cause damage to multiple systems, especially the male reproductive system, and testis is an important target organ. Oxidative stress injury is the core mechanism of spermatogenesis disorder caused by DEHP. DEHP exposure can cause oxidative stress or reactive oxygen species (ROS) increase in germ cells, and on this basis, promote cell apoptosis or cause excessive autophagy. The toxicity of DEHP to Leydig cells is mainly to interfere with the synthesis of steroid hormones. For Sertoli cells, ferroptosis and destruction of the blood-testis barrier are common injury mechanisms. In addition, gene methylation caused by DEHP not only affects the spermatogenic process, but also has epigenetic effects on offspring. In this paper, we reviewed the pathological damage, germ cell toxicity and epigenetic effects of DEHP on testis, and focused on the damage and molecular mechanism on testicular spermatogenic cells, Leydig cells and Sertoli cells. Future research is required to elucidate the body’s clearance mechanism and treatment plan after exposure to DEHP and whether DEHP will damage the function of myoid cells. It is hoped that this can provide new ideas for prevention and treatment of male reproductive disorders resulting from long-term exposure to plastic products.
6.Construction of luminescent bacteriophage using CRISPR technology and its application in Escherichia coli indentification
Minwei LI ; Jing YAN ; Hangyi LI ; Zhiyun HAO ; Zhong NI ; Zhaoyang HU ; Xiaorong WANG ; Menghan XU ; Chi WANG ; Ruibing LI ; Chengbin WANG
Chinese Journal of Laboratory Medicine 2024;47(4):437-443
Objective:To construct a recombinant bioluminescent bacteriophage (HT7) targeting Escherichia coli, and evaluate its ability to identify Escherichia coli. Methods:Initially, pCRISPR-sg (1-10) and PFN-1000 plasmid strains were constructed by genetic engineering, and the most efficient small guild RNA (sgRNA) were screened by bilayer plate. By the gene editing technique, which comprised homologous recombination and clustered regularly interspaced short palin dromic repeats (CRISPR)-Cas system, the Nanoluc luciferase gene was integrated into the downstream non-coding region of 10A gene of T7 phage, to constructe the bioluminescent phage HT7 successfully. The difference of biological characteristics between HT7 phage and T7 phage was evaluated by plaque assay and liquid amplification assay. In addition, 51 strains of Escherichia coli, 20 strains of Klebsiella pneumoniae, 14 strains of Staphylococcus aureus, 6 strains of Enterococcus faecium, 5 strains of Enterococcus faecalis, 3 strains of Acinetobacter baumannii and 1 strain of Pseudomonas aeruginosa were collected and isolated to evaluate the limit of detection and specificity of HT7 phage. Results:Among the 10 CRISPR-targeted cleavage systems constructed, sgRNA8 exhibited the highest cleavage efficiency, with a cleavage rate of 0.18. After three rounds of recombination screening using the pCas9/pCRISPR/PFN-1000 triple-plasmid system, PCR validation yielded recombinant phage bands at 2 798 bp, indicating the successful construction of the HT7 phage. The recombinant phage showed significant differences in biological characteristics in terms of lysis efficiency ( P<0.001), one-step growth curve ( P=0.001), and infection multiplicity ( P=0.031). Both lysis burst time and log growth node were extended by 10 min, with the optimal infection multiplicity being 0.1. Clinical sample testing identified lysis of 6 strains of Escherichia coli within 4.5 h, while other strains remained unaffected, with detection of pathogenic bacteria below 10 CFU/ml. Conclusions:The developed pCas9/pCRISPR/PFN-1000 triple-plasmid editing system efficiently edits the bacteriophage genome. The constructed HT7 fluorescent bacteriophage enables the detection of Escherichia coli below 10 CFU/ml within 4.5 hours, demonstrating low detection limits and high detection specificity.
7.Application of Allograft Endometriosis Rat Model in Pharmaco-dynamic Evaluation of GnRH Agonists
Ruihua ZHONG ; Guoting LI ; Wenjie YANG ; Xiangjie GUO ; Jieyun ZHOU ; Yingyi HU ; Qicheng NI ; Ye YANG ; Min ZHANG ; Yan ZHU
Laboratory Animal and Comparative Medicine 2024;44(2):127-138
Objective To establish an allogeneic rat model of endometriosis and to evaluate the effects of gonadotropin-releasing hormone (GnRH) agonist GenSci006 on experimental rat endometriosis. Methods Endometrium from SPF grade donor female SD rats were transplanted onto the abdominal wall of recipient female rats to construct an allogeneic endometriosis model. The rats undergoing sham surgery were divided into the sham group. Three weeks later, the length, width and height of the ectopic endometrium were measured, and the volume of the endometrium (V1) was calculated before drug administration. The modeling rats were randomly divided into four groups: model group, triptorelin group (0.25 mg/kg), GenSci006-1 group (0.125 mg/kg) and GenSci006-2 group (0.25 mg/kg). Each group had 16 rats and received a single dose of the corresponding drug. The sham group and model group were administered an equal volume of solvent. Three weeks after administration, ectopic endometrium was measured to calculate the volume V2 and inhibition rate. The effect of GenSci006 on rat uterus and ovarian tissues was assessed by comparing organ coefficients and changes in pathological sections. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of serum estradiol (E2), progesterone (P4), follicle stimulating hormone (FSH), and luteinizing hormone (LH). Real-time fluorescent quantitative PCR was used to detect the expression of GnRH receptor (GnRHR) mRNA in the hypothalamus and pituitary. Western blot was used to detect the expression of estradiol receptor alpha (ERα), beta (ERβ) and progesterone receptor (PR) in ectopic endometrium. Results Three weeks after administration, compared with the model group, the body weight of rats in the triptorelin and GenSci006-2 groups significantly increased (P < 0.05), while the volume of ectopic endometrium significantly decreased (P < 0.05). Compared with the sham group, the model group showed no significant changes in uterine and ovarian organ coefficients or endometrial thickness (P > 0.05). Compared with the model group, the uterine organ coefficients and endometrial thickness were significantly reduced in the triptorelin and GenSci006-2 groups (P < 0.05). Compared with the sham group, the serum levels of E2, P4, FSH and LH in the model group showed no significant changes (P > 0.05). Compared with the model group, the ovarian organ coefficient and serum P4 levels of rats in the Triptorelin, GenSci006-1, and GenSci006-2 groups were significantly reduced (P < 0.05), while the serum LH levels of rats in the GenSci006-1 group were significantly increased (P < 0.05). However, there were no significant changes in serum E2 and FSH levels in each group (P > 0.05). Compared with the model group, the expression levels of GnRHR mRNA in the pituitary tissue of rats in the triptorelin and GenSci006-2 groups were significantly downregulated (P < 0.05), with no significantly changes in the hypothalamus (P > 0.05). There were no significant changes in the expression level of GnRHR mRNA in the hypothalamus or the protein levels of ERα, ERβ and PR in the ectopic endometrial tissue in any group (P > 0.05). Conclusion The allogeneic endometriosis rat model is a suitable animal model for screening and evaluating drugs for treating endometriosis. The volume of ectopic endometrium, inhibition rate, uterine and ovarian organ coefficients, and serum E2 levels may serve as indicators for detecting drug efficacy.
8.Downregulation of MUC1 Inhibits Proliferation and Promotes Apoptosis by Inactivating NF-κB Signaling Pathway in Human Nasopharyngeal Carcinoma
Shou-Wu WU ; Shao-Kun LIN ; Zhong-Zhu NIAN ; Xin-Wen WANG ; Wei-Nian LIN ; Li-Ming ZHUANG ; Zhi-Sheng WU ; Zhi-Wei HUANG ; A-Min WANG ; Ni-Li GAO ; Jia-Wen CHEN ; Wen-Ting YUAN ; Kai-Xian LU ; Jun LIAO
Progress in Biochemistry and Biophysics 2024;51(9):2182-2193
ObjectiveTo investigate the effect of mucin 1 (MUC1) on the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) and its regulatory mechanism. MethodsThe 60 NPC and paired para-cancer normal tissues were collected from October 2020 to July 2021 in Quanzhou First Hospital. The expression of MUC1 was measured by real-time quantitative PCR (qPCR) in the patients with PNC. The 5-8F and HNE1 cells were transfected with siRNA control (si-control) or siRNA targeting MUC1 (si-MUC1). Cell proliferation was analyzed by cell counting kit-8 and colony formation assay, and apoptosis was analyzed by flow cytometry analysis in the 5-8F and HNE1 cells. The qPCR and ELISA were executed to analyze the levels of TNF-α and IL-6. Western blot was performed to measure the expression of MUC1, NF-кB and apoptosis-related proteins (Bax and Bcl-2). ResultsThe expression of MUC1 was up-regulated in the NPC tissues, and NPC patients with the high MUC1 expression were inclined to EBV infection, growth and metastasis of NPC. Loss of MUC1 restrained malignant features, including the proliferation and apoptosis, downregulated the expression of p-IкB、p-P65 and Bcl-2 and upregulated the expression of Bax in the NPC cells. ConclusionDownregulation of MUC1 restrained biological characteristics of malignancy, including cell proliferation and apoptosis, by inactivating NF-κB signaling pathway in NPC.
9.CT and MRI findings of different types of small round cell tumor in the nasal cavity and sinuses
Bo QIAN ; Yeming ZHONG ; Ting NI ; Hongbo JI ; Jie CUI ; Zigang CHE
Journal of Practical Radiology 2024;40(4):539-542
Objective To investigate the CT and MRI findings of different types of small round cell tumor(SRCT)in the nasal cavity and sinuses.Methods A retrospective analysis was conducted on the imaging data and clinical data of 35 SRCT patients confirmed by pathology.Thirty-one SRCT patients underwent CT examination,and 19 SRCT patients underwent MRI examination.Results There were 20 cases of tumors that invaded the nasal cavity and 19 cases involved the sinuses,including 11 cases of the maxillary sinus,7 cases of the ethmoid sinus,2 cases of the sphenoid sinus,and 1 case of the frontal sinus.CT findings of SRCT were all soft tissue masses.Lymphoma was relatively homogeneous with mild bone destruction,and usually involved nasal vestibular skin.Rhabdomyosarcoma(embryonic type)happened at an early age and easily caused bone destruction and metastasis.Melanoma generally occurred in the nasal septum and nasal cavity,which was prone to bleeding.Small cell neuroendocrine carcinoma was heterogeneous,with moderate to significant enhancement,bone absorption and destruction were often noticed.The MRI manifestations of SRCT were equal or long signal on T1WI,high signal on T2WI,and significant diffusion limitation on diffusion weighted imaging(DWI)and apparent diffusion coefficient(ADC)except for melanoma.On contrast-enhanced images,lymphoma showed mild to moderate enhancement,rhabdomyosarcoma showed typical"grape sign",and small cell neuroendocrine carcinoma showed"sieve"and"map-like"obvious enhancement.Typical melanoma showed a high signal on T1WI and a low signal on T2WI and usually caused bleeding.The MRI findings were related to the presence of melanoma and hemorrhage within the lesion.Conclusion SRCT of the nasal cavity and sinuses have a high degree of malignancy and poor prognosis,CT and MRI have many similar manifestations.Combining clinical data,bone destruction,MRI enhancement,and DWI sequence can effectively distinguish different types of SRCT,as well as squamous cell carcinoma of the nasal cavity and sinuses and adenoid cystic carcinoma.
10.Quality Standard of Tibetan Medicine "Yajima" (Chrysosplenium Axillare)
Gang REN ; Chaowei PU ; Jingjing WEN ; Wei JIANG ; Guoyue ZHONG ; Weizao LUO ; Zhen NI ; Jiamei XIANG
Chinese Journal of Modern Applied Pharmacy 2024;41(4):469-475
OBJECTIVE
To establish the quality standards of medicinal materials in light of related methods in the general principles of part four of Chinese Pharmacopoeia(2020 Edition), and to conduct systematic research on the Tibetan medicine "Yajima"(Chrysosplenium axillare).
METHODS
The powder characteristics of medicinal materials were described by microscopic identification method. Silica gel GF254 thin-layer plate was employed to establish a TLC identification method with 5-O-demethylapulein and oxyayanin A as reference substances. Loss on drying, total ash, acid-insoluble ash and ethanol-soluble extractives of 10 batches of Chrysosplenium axillare were determined according to the general principles of part four of Chinese Pharmacopoeia(2020 Edition). HPLC was used to establish the characteristic chromatogram of Chrysosplenium axillare, and the content determination method was established with chrysosplenoside I(CI) and chrysosplenoside A(CA) as the quality control index components of Chrysosplenium axillare.
RESULTS
The water content, total ash, acid-insoluble ash, ethanol-soluble extractive and the content of CI and CA of all samples varied in the ranges of 9.17%−12.52%, 14.11%−16.74%, 1.50%−4.72%, 32.77%−40.30%, 0.30%−0.99% and 0.28%−0.88%, respectively.
CONCLUSION
The identification and content determination methods of Yajima(Chrysosplenium axillare) are established for the first time. The methods are easy to operate and exclusive, which is of great significance to accurately evaluate the internal quality of medicinal materials and ensure the quality of drug used.


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