1.Effect mechanism of LncRNA MALAT1 on doxorubicin resistance in osteosarcoma cells
Fudong LIANG ; Shufang DI ; Wei LUO ; Jianghua QI ; Libing LIU
China Pharmacy 2025;36(6):698-703
OBJECTIVE To investigate the relationship of long non-coding RNA (LncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) and doxorubicin (DOX) resistance in osteosarcoma (OS) cells. METHODS MG-63 and MG-63/DOX cells were treated with different concentrations of DOX (0, 0.01, 0.05, 0.1, 1 μmol/L), and survival rates and half maximal inhibitory concentration were determined using CCK-8 assay. The expressions of LncRNA MALAT1 in MG-63 and MG-63/ DOX cells were detected by real-time quantitative fluorescence PCR. MG-63/DOX cells were divided into Control group, knocking down LncRNA MALAT1 negative control (sh-NC) group, sh-MALAT1 group, sh-MALAT1+anti-NC group, and sh-MALAT1+ anti-miR-154-5p group. The expressions of LncRNA MALAT1, miR-154-5p and cyclin D1 (CCND1) mRNA in MG-63/DOX cells of each group were detected. The effects of knocking down LncRNA MALAT1 on the proliferation, migration, invasion, and apoptosis of MG-63/DOX cells were detected by CCK-8 assay, scratch test, Transwell experiment and flow cytometry, respectively. The expression of proliferating cell nuclear protein (PCNA) and CCND1 protein in MG-63/DOX cells was detected by Western blot assay. Interactions between LncRNA MALAT1 and miR-154-5p, miR-154-5p and CCND1 were detected by dual luciferase reporter gene experiment. RESULTS Compared with 0 μmol/L DOX, 0.01, 0.05, 0.1 and 1 μmol/L DOX could reduce the survival rates of MG-63 and MG-63/DOX cells (except for 0.01 μmol/L DOX) (P<0.05), IC50 were 0.07 and 0.13 μmol/L, respectively. The survival rate, cell migration number and invasion number of MG-63/DOX cells, scratch closure rate, mRNA expressions of LncRNA MALAT1, mRNA and protein expressions of CCND1, and PCNA protein expression in sh-MALAT1 group were significantly lower than sh-NC group and Control group; the apoptosis rate and miR-154-5p expression were significantly higher than sh-NC group and Control group (P<0.05). sh-MALAT1+anti-miR-154-5p group was able to reverse the aforementioned biological effects in sh-MALAT1 group (P<0.05). In MG-63/DOX cells transfected with both MALAT1-wild type (WT) and CCND1-WT, the luciferase activity in the miR-154-5p mimic group was significantly lower than mimic negative control group (P< 0.05). CONCLUSIONS Knocking down LncRNA MALAT1 can inhibit the DOX resistance of OS cells, and its mechanism may be targeting the miR-154-5p/CCND1 axis.
3.Research progress on quality control methods for monitoring illicit drugs use in wastewater
Yue XIAO ; Shuai YUAN ; Ruxin LUO ; Ruiqin ZHU ; Bin DI ; Ping XIANG
Journal of China Pharmaceutical University 2025;56(2):139-147
The use of wastewater analysis, or wastewater-based epidemiology, to assess and monitor the situation of drug abuse is now widely used at home and abroad. However, there is currently a lack of effective evaluation methods and effective ways of comparison, supervision and standardization, which is not conducive to the analysis and comparisons of data in different countries and regions. Quality control techniques can control the laboratory's analytical errors, safeguard the consistency and comparability of identification conclusions, and promote the further improvement of the level and capacity of urban drug governance, thus playing significant roles. This paper provides an overview of sample collection, sample preservation and transportation, laboratory analysis, back-calculation of drug use and external laboratory quality control in the process of wastewater analysis, with a view to exploring more comprehensive scientific and objective methods and approaches suitable for examining and evaluating qualitative and quantitative analysis of drugs in wastewater among laboratories.
4.Methods for Identifying Seeds of Chinese Medicinal Materials: A Review
Zhihao LIU ; Liangping ZHA ; Li LUO ; Yuyang ZHAO ; Xiaolin LI ; Yuan YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):1-11
Seeds are the source for the production of Chinese medicinal materials. The seed authenticity and quality of directly affect the effectiveness and safety of Chinese medicinal materials. The seed quality is faced with the problems such as mixed sources, existence of adulterants and seeds stocked for years, low maturity, and low purity. To ensure the high-quality and sustainable development of the Chinese medicinal material industry, it is urgent to standardize the seed market and identify and evaluate the quality of the seeds circulating in the market. Seed identification methods include visual inspection, microscopic observation, micro-character identification, chemical fingerprinting, molecular identification, electronic nose, X-ray diffraction, electrochemical fingerprinting, spectral imaging, and artificial intelligence. These methods have different application scopes and unique advantages and disadvantages. According to the different species of Chinese herbal medicines and different requirements of testing sites, suitable methods can be selected to achieve rapid and accurate identification with low costs. In the future, the seed identification methods should be developed based on emerging technologies with interdisciplinary knowledge, and intelligent, nondestructive, and single-grain detection methods are needed for the modern Chinese medicinal material industry. This paper introduces the seed identification technologies currently applied in research and production, compares the principles, applicability, advantages, and disadvantages of different technologies, and provides an outlook on the future development of seed identification technologies, aiming to provide a reference for the identification and quality evaluation of seeds of Chinese medicinal material.
5.Specific PCR for Identification of Astragalus membranaceus var. mongholicus Seeds, A. membranaceus Seeds, and Adulterants
Li LUO ; Li HU ; Chao JIANG ; Ziyuan CHEN ; Xiaolin LI ; Yuan YUAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):21-28
ObjectiveTo establish a method based on specific polymerase chain reaction (PCR) that can accurately and rapidly identify Astragalus membranaceus var. mongholicus (AMM) seeds and A. membranaceus (AM) seeds. MethodThe Chloroplast Genome Information Resource (CGIR) and IdenDSS were used to obtain the characteristic DNA fragments of AMM and AM, and the specific single nucleotide polymorphism (SNP) sites of AMM and AM were screened out, on the basis of which the specific primers MG-F/MG-R of AMM and MJ-F/MJ-R of AM were designed. The specific PCR method for identifying AMM and AM was established and optimized, and the specificity and applicability of the method were investigated. ResultThe specific PCR conditions for the identification of AMM were primers MG-F/MG-R, annealing at 62 ℃, and 28 cycles. After PCR amplification and gel electrophoresis, the specific band appeared at about 220 bp, with no band for the seeds of AM or adulterants. The specific PCR conditions for identifying the AM were primers MJ-F/MJ-R, annealing at 58 ℃, and 28 cycles. After PCR amplification and gel electrophoresis, the band appeared at about 150 bp, with no band of AMM or adulterants. ConclusionThe specific PCR method established in this study can accurately and quickly identify the seeds of AMM and AM, which provides a basis for the classification and accurate identification of Astragalus seeds and adulterants.
6. Research progress in RIPK3 and its inhibitors
Di CHEN ; Jun PENG ; Xiu-Ju LUO
Chinese Pharmacological Bulletin 2024;40(3):401-405
Receptor-interacting serine/threonine-protein kinase 3(RIPK3),a member of the RIP kinase family,plays an important role in cell death,especially in necroptosis. In addition,RIPK3 is also involved in apoptosis and pyroptosis,suggesting that RIPK3 may be the intersection of multiple cell death and it possesses the potential to be a target for precise regulation of cell death. According to the kinase binding mode,current RIPK3 inhibitors can be classified into type ,type Ⅱ and other types. This review summarizes the research progress in the role of RIPK3 in cell death and its inhibitors,which is of great significance in seeking drugs for the treatment of injury-related diseases.
7.Factor analysis of pain after arthroscopic rotator cuff suture bridge
Li-Na SI ; Jin-Wei LUO ; Di WU ; Yue-Bing QIAO ; Yong-Ming LÜ ; Cong XU
Acta Anatomica Sinica 2024;55(2):210-214
Objective To analyze the factors associated with pain after arthroscopic rotator cuff bridge suture.Methods According to the inclusion and exclusion criteria,the data of 112 patients with unilateral rotator cuff injury who received arthroscopic bridge suture in our department were collected and were investigated in the form of telephone follow-up.In this study,SPSS 23.0 was used to input data and conduct statistical analysis.Logistic regression analysis was used to analyze the correlation between the above influencing factors and postoperative pain.Results A total of 112 patients were included for statistical analysis,single factor analysis revealed,including course of disease,smoking history,preoperative University of California,Los Angeles(UCLA)score,Constant score,numeric rating scale(NRS),size of rotator cuff tear,whether it was full-thickness tear and degree of tendon retraction might be related to postoperative pain(P<0.05).The age,gender,body mass index(BMI),drinking history,diabetes and hypertension were not related to postoperative pain(P>0.05).Multiple linear regression analysis concluded that there were four factors related to postoperative pain,and the correlation degree was preoperative NRS,preoperative UCLA score,tear size and smoking history.Conclusion The causes of postoperative pain after arthroscopic rotator cauff repair are complex and diverse.Analyzing the cause of postoperative pain can effectively reduce the pain of patients and promote the recovery of shoulder joint function.
8.Overview of Research on Intervention Methods in the Mechanism of Acupuncture Treatment of Cocaine Addiction
Luqiang SUN ; Fangli LUO ; Di QIN ; Shuai CHEN ; Haiyan WANG ; Ying LI
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(1):275-280
Cocaine,as a widely abused and highly addictive drug,has a serious impact on the physical and mental health of individuals and carries a certain degree of social harm and economic burden.Acupuncture can assist in the treatment of cocaine addiction with fewer side effects.However,a well-defined mode of stimulation is an important factor in elucidating the various mechanisms by which acupuncture treats disease.This paper summarizes the problems in the mechanism of cocaine addiction,such as different parameters of stimulation,unstable depth of acupuncture,different acupoint selection,and different lengths of acupuncture time.To standardize the intervention measures of acupuncture experiments,it is suggested that in future research,the stimulation method should explore the best parameters,the selection of acupoints should be based on clinical practice,the timing of acupuncture should be objective,and the treatment course should consider the effects of acupuncture.
9.Stepwise treatment strategy for spontaneous osteonecrosis of the medial femoral condyle of the knee joint
Jianke PAN ; Meiping YANG ; Yanhong HAN ; Di ZHAO ; Hetao HUANG ; Houran CAO ; Jun LIU ; Minghui LUO ; Xiang LI ; Hongyun CHEN ; Weiyi YANG
Chinese Journal of Tissue Engineering Research 2024;28(12):1907-1913
BACKGROUND:Currently,there have been a variety of conservative and surgical treatment plans for spontaneous osteonecrosis of the knee,achieving excellent results.However,a broad consensus on indication and guide of surgical treatment has not been announced.In clinical practice,there is still a misunderstanding that unicondylar replacement or total knee arthroplasty should be performed upon the discovery of spontaneous osteonecrosis of the knee,while an urgent need for universal access to the concept of stepwise therapy. OBJECTIVE:To summarize and find the factors leading to the poor effect of conservative treatment in spontaneous osteonecrosis of the knee,which occurred on the medial femoral condyle,from the literature and clinical cases,at the same time,combined with the Koshino stage,to propose the strategy of stepwise spontaneous osteonecrosis of the knee treatment on the medial femoral condyle. METHODS:A systematic search of the literature database was conducted to summarize the factors leading to poor outcomes of conservative treatment in spontaneous osteonecrosis of the medial femoral condyle.Meanwhile,according to the Clinical&Health Records for analytics&Sharing system,the cases receiving conservative and surgical treatment in spontaneous osteonecrosis of the medial femoral condyle in the Department of Orthopedics of Guangdong Provincial Hospital of Chinese Medicine from January 2017 to January 2023 were analyzed retrospectively,then the causes of success and failure in typical cases were summarized and analyzed. RESULTS AND CONCLUSION:(1)Early diagnosis and treatment of spontaneous osteonecrosis of the knee were very important for prognosis.For sudden knee pain in some patients,if no obvious abnormality was found in the X-ray examination,and the symptoms persisted and could not be relieved for more than 1 week,an MRI examination was recommended to detect early spontaneous osteonecrosis of the knee.(2)The X-ray images of Koshino stage 1 and stage 2 of spontaneous osteonecrosis of the medial femoral condyle were difficult to be distinguished,which needed to be probed by MRI.MRI images of Koshino stage 1 were mainly characterized by bone marrow edema,and an osteonecrosis area with a clear boundary was not formed,while MR images of Koshino stage 2 showed a necrotic area with a clear boundary.(3)Five factors leading to the poor effect of conservative treatment on spontaneous osteonecrosis of the medial femoral condyle were summarized:a.The necrotic area was>5 cm2;b.The necrotic area accounted for more than 40%of the condyle;c.relative compression percentage of medial meniscus≥33%(with or without medial meniscus injury and subchondral bone marrow edema);d.MRI depth of necrotic area(anterior-posterior diameter of sagittal necrotic area)>20 mm;e.varus deformity of lower limb>6°.(4)Conservative treatment of spontaneous osteonecrosis of the knee in Koshino stage 1 was good.For spontaneous osteonecrosis of the knee in Koshino stage 2,conservative treatment was preferred or combined with drilling decompression.If there was no relief or improvement of symptoms or in MRI after 3 months,while the patient had any of the previous five factors,then knee preservation surgery should be considered.For spontaneous osteonecrosis of the knee in Koshino stage 3 and stage 4,knee preservation surgery should be selected based on the previous five factors,including age,gender and activity level of the patient.Total knee arthroplasty was used for spontaneous osteonecrosis in Koshino stage 4,which was associated with symptomatic patellofemoral arthritis,valgus alignment,or necrotic area,which greatly affected the stability of unicondyle prosthesis.
10.Analyses of high-risk factors for poor neurologic prognosis in full-term neonatal purulent meningitis based on clinical and MRI characteristics
Huanyu LUO ; Di HU ; Shuangfeng YANG ; Hang LI ; Yun PENG
Chinese Journal of Radiology 2024;58(3):301-306
Objective:To analyze high-risk factors for poor neurological prognosis in full-term neonatal purulent meningitis based on clinical and brain MRI features.Methods:This study was a case-control study. The clinical and brain MRI data of 79 neonates with purulent meningitis were retrospectively collected at Beijing Children′s Hospital, Capital Medical University from January 2016 to January 2022. Follow-up assessments including growth and development, as well as neurological sequelae, were conducted over a minimum follow-up period of 6 months. The patients were divided into two groups with good ( n=49) and poor prognosis ( n=30) according to follow-up results. Chi-square tests were used to compare clinical and brain MRI features between the two groups, and a multivariate logistic regression analysis was performed to explore the high-risk factors for poor neurologic prognosis in full-term neonates with purulent meningitis. Results:There were statistically differences between two groups regarding the incidence of seizures, early-onset manifestations, positive cerebrospinal fluid (CSF) culture, CSF white cell counts, and CSF protein concentration ( P<0.05). Statistically differences were also found in the occurrence rates of ependymitis, obvious ventricular dilatation/hydrocephalus, spotty and patchy brain injury/hemorrhage, and destructive lesions within the brain parenchyma ( P<0.05). The results of multivariate logistic regression analysis indicated that seizures ( OR=5.722, 95% CI 1.126-29.072, P=0.035), early-onset neonatal purulent meningitis ( OR=3.657, 95% CI 1.073-12.459, P=0.038), ependymitis ( OR=8.851, 95% CI 1.169-67.017, P=0.035), obvious ventricular dilatation/hydrocephalus ( OR=12.675, 95% CI 1.085-148.110, P=0.043), and destructive lesions within the brain parenchyma ( OR=16.370, 95% CI 1.575-170.175, P=0.019) were independent risk factors for poor prognosis. Conclusions:The occurrence of seizures, early-onset manifestations as well as ependymitis, obvious ventricular dilatation/hydrocephalus, and destructive lesions within the brain parenchyma on MRI are high-risk factors for poor prognosis in the full-term neonate with purulent meningitis.

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