1.Development status analysis and suggestions of TCM pharmacists in Chinese public TCM hospitals
Baojuan XUE ; Ning WU ; Yang ZHAO ; Junshu GE ; Yi WANG ; Zheyuan LIU ; Zhaoheng YANG ; Ying SUN
China Pharmacy 2025;36(8):903-907
OBJECTIVE To understand the development status and existing problems of traditional Chinese medicine(TCM) pharmacists in public TCM hospitals in China, aiming to provide suggestions for the competent departments to formulate management policies for TCM pharmacists and promote the healthy development of TCM. METHODS The data on the number and professional titles of TCM pharmacists in public TCM hospitals in China from 2019 to 2023 were collected. Descriptive analysis was employed to analyze the number, distribution and professional titles of TCM pharmacists in public TCM hospitals across the country, and to measure the quantity shortfalls of the number of TCM pharmacists in these hospitals. RESULTS From 2019 to 2023, the number of TCM pharmacists in public TCM hospitals in China grew slowly, with an average annual growth rate of 2.56%. However, the proportion of TCM pharmacists to the total number of pharmacists in public TCM hospitals gradually decreased, with an average annual growth rate of -0.65%. In terms of hospital grades, the number of TCM pharmacists in tertiary public TCM hospitals showed positive growth, while those in secondary and primary public TCM hospitals showed negative growth. In terms of hospital types, the average annual growth rate of TCM pharmacists in TCM hospitals was 2.22%, in integrated Chinese and Western medicine hospitals it was 7.97%, and in ethnic minority medicine hospitals it was 2.74%. The development of TCM pharmacists in different provinces was uneven. The annual growth rate of TCM pharmacists in Guizhou exceeded 10%, while the growth rate in Hunan and Heilongjiang was negative. In 2023, the number of TCM pharmacists per thousand population in public TCM hospitals was 0.03, indicating a relatively low staffing level. The professional titles of TCM pharmacists in public TCM hospitals were mainly primary and E-mail:601907549@qq.com intermediate, with a total of 67.33%. According to the calculation that the proportion of TCM pharmacists to pharmacists was not less than 60%, public TCM hospitals and hospitals of integrated TCM and Western medicine should be reconfigured with TCM pharmacists 6 212 and 1 288 people, respectively. CONCLUSIONS The number of TCM pharmacists in public TCM hospitals is growing slowly, with insufficient staffing levels, relatively low professional titles, and uneven distribution and development across provinces. It is suggested that relevant competent departments strengthen policy guidance, increase the attention given by the state level to TCM pharmacists, strengthen the construction of the talent team for TCM pharmacists, improve the quality and optimize the allocation of TCM pharmacist talents in order to promote the high-quality development of TCM services.
2.Analysis of the nonlinear relationship between hypothermic machine perfusion parameters and delayed graft function and construction of an optimized predictive model based on sampling algorithms
Boqing DONG ; Chongfeng WANG ; Yuting ZHAO ; Huanjing BI ; Ying WANG ; Jingwen WANG ; Zuhan CHEN ; Ruiyang MA ; Wujun XUE ; Yang LI ; Xiaoming DING
Organ Transplantation 2025;16(4):582-590
Objective To analyze the nonlinear relationship between hypothermic machine perfusion (HMP) parameters and delayed graft function (DGF) and optimize the construction of a predictive model for DGF. Methods The data of 923 recipients who underwent kidney transplantation from deceased donors were retrospectively analyzed. According to the occurrence of DGF, the recipients were divided into DGF group (n=823) and non-DGF group (n=100). Donor data, HMP parameters and recipient data were analyzed for both groups. The nonlinear relationship between HMP parameters and the occurrence of DGF was explored based on restricted cubic splines (RCS). Over-sampling, under-sampling and balanced sampling were used to address the imbalance in the proportion of DGF to construct logistic regression predictive models. The area under the curve (AUC) of each model was compared in the validation set, and a nomogram model was constructed. Results Donor BMI, cold ischemia time of the donor kidney, and HMP parameters (initial and final pressures, resistance, and perfusion time) were significantly different between the DGF and non-DGF groups (all P<0.05). The RCS analysis revealed a threshold-like nonlinear relationship between HMP parameters and the risk of DGF. Among the models constructed using different sampling methods, the balanced sampling model had the highest AUC. Using this model, a nomogram was constructed to stratify recipients based on risk scores. Recipients in the high-risk group had higher serum creatinine levels at 1, 6, and 12 months after kidney transplantation compared to those in the low-risk group (all P<0.05). Conclusions There is a nonlinear relationship between HMP parameters and the risk of DGF, and the threshold is helpful for organ quality assessment and monitoring of graft function after transplantation. The predictive model for DGF constructed on the base of balanced sampling algorithms helps perioperative decision-making and postoperative graft function monitoring of kidney transplantation.
3.Effects of Angelica Sinensis and Astragalus capsules on myocardial autophagy in rats with heart failure
Xue WU ; Xin-Fang LÜ ; Xiao-Dong ZHI ; Xin-Ke ZHAO ; Ying-Dong LI
The Chinese Journal of Clinical Pharmacology 2024;40(10):1453-1457
Objective To investigate whether Angelica Sinensis and Astragalus capsules(AAC)regulates myocardial autophagy in heart failure rats via the phosphatidylinositol 3 kinase(PI3K)/protein kinase(Akt)/mammalian target of sirolimus(mTOR)signaling pathway.Methods A rat model of heart failure was constructed by intraperitoneal 2.5 mg·kg-1 doxorubicin,and another 8 rats served as the control group.The modeling rats were randomly divided into model group,control group and experimental-L,-M,-H groups.Control group was given 30 mg·kg-1 3-methyladenine by intraperitoneal injection;experimental-L,-M,-H groups were given 150,300 and 450 mg·kg-1 AAC by gavage,respectively;blank and model groups were given the same quantity of sterile distilled water.Six groups were administered once daily for 6 weeks.The cardiac function was measured by ultrasound,and the expression levels of PI3K,Akt,mTOR,sequestosome 1(P62)and microtubule-associated light chain protein 3-Ⅱ/Ⅰ(LC3 Ⅱ/Ⅰ)in myocardial tissue were measured by Western blot.Results In the blank,model,control and experimental-H groups,the left ventricular ejection fraction values were(85.00±3.63)%,(56.75±4.83)%,(75.63±3.70)%and(72.75±4.23)%;the relative expression levels of PI3K were 1.00±0,0.28±0.05,0.64±0.08 and 0.74±0.16;phosphorylated Akt/Akt were 1.00±0,0.49±0.06,0.90±0.16 and 0.95±0.10;phosphorylated mTOR/mTOR values were 1.00±0,0.42±0.09,0.73±0.13 and 0.83±0.08;the relative expression levels of P62 proteins were 1.00±0,0.24±0.12,0.57±0.09 and 0.96±0.10;the relative expression levels of LC3 Ⅱ/Ⅰ proteins were 1.00±0,4.31±0.75,2.20±0.76 and 1.59±0.24,respectively.Compared to the model group,statistical significant were identified in the experimental-H and control groups(all P<0.05).Conclusion AAC can regulate PI3K/Akt/mTOR pathway,inhibit myocardial autophagy and improve cardiac function in rats with heart failure.
4.Role of sphingosine-1-phosphate signaling in the proliferation of breast cancer BT549 cells
Juan SONG ; Ming WANG ; Xin-Yang LIU ; Hao-Tian ZHANG ; Qi ZHANG ; Xue-Mei ZHAO ; Ying LIU ; Hong-Xia CUI
The Chinese Journal of Clinical Pharmacology 2024;40(11):1578-1582
Objective To study the role of sphingosine-1-phosphate(S1P)signal on the proliferation of breast cancer BT549 cells.Methods Cells were divided into control group and experimental group,experimental group were treated with 0.1,1.0,10.0 μmol·L-1 S1P receptor agonist SEW2871 for 72 h.Control group was cultured with 0.1%fetal bovine serum.Cell proliferation was detected by methyl thiazolyl tetrazolium(MTT)assay.Cell models of overexpressing S1P receptors in BT549 were divided into three groups:blank plasmid group(LUC),wild type S1P receptor overexpression group(WT),S1P receptor phosphorylation site mutation overexpression group(MUT);the proliferation ratio was detected by MTT,the number of cell clones was counted by colony formation experiment.S1P antagonist W146(10 μmol·L-1)and protein kinase(AKT)signaling inhibitor MK2206(90 nmol·L-1)were used to detect the role of S1P signaling in the proliferation of breast cancer cells.The expression of phosphorylate signal transducer and activator of transcription 3(p-STAT3),c-Myc proteins were detected by Western blot.Results The growth ratio of BT549 cells in control group and 0.1,1.0,10.0 μmol·L-1experimental groups were 1.00±0.03,1.13±0.06,1.06±0.10 and 1.07±0.03,0.1 μmol·L-1 SEW2871 promot the cell proliferation(P<0.05).Compared between WT group,MUT group and LUC group,the growth rate and the number of clonal colonies were increased after overexpression of S1P receptor(all P<0.05).The growth ratio of BT549 cells after treatment with W146 and MK2206 in the LUC group,WT group and MUT group were 1.25±0.12,1.31±0.03,1.43±0.14 and 0.87±0.15,0.77±0.03,0.88±0.02.Compared between MUT group,WT group and corresponding DMSO group,the differences were statistically significant(all P<0.01).The number of cell clony formation number after treatment with W146 were 65.65±5.12,141.48±5.63 and 93.64±5.14;compared between MUT,WT group and corresponding DMSO group,the differences were statistically significant(all P<0.05).The relative protein expression levels of p-STAT3 in LUC group,WT group and MUT group were 0.67±0.04,0.69±0.08 and 0.81±0.06,the relative protein expression levels of proto-oncogene c-Myc were 1.69±0.03,0.70±0.10 and 0.67±0.07,compared between WT group,MUT group and corresponding DMSO group,the difference was statistically significant(P<0.05).Conclusion S1P signaling can promote proliferation in breast cancer BT549 cells,and the mechanism could be related to AKT and STAT3 signaling pathway.
5.Meta-analysis of the effects of SLCO1B1 gene polymorphisms on the efficacy and safety of rosuvastatin
Chunyun LU ; Song WANG ; Kefeng LIU ; Ying XUE ; Juanjuan CHEN ; Yuanxia ZHAO ; Shuzhang DU
China Pharmacy 2024;35(19):2397-2403
OBJECTIVE To study the correlation between SLCO1B1 (521T>C and 388A>G) gene polymorphisms and the efficacy and safety of rosuvastatin. METHODS Retrieved from PubMed, Embase, Cochrane Library, PharmGKB, CNKI database and Wanfang database, the studies about the effects of 521T>C and 388A>G gene polymorphisms on the efficacy and safety of rosuvastatin were collected during the inception to Dec. 2023. The included data were analyzed by using RevMan 5.3 software. RESULTS A total of 16 studies were included. The results of meta-analysis showed that 521T>C gene polymorphism was significantly correlated with the efficacy of rosuvastatin. In the dominant gene model, compared with TT genotype, CC+TC genotype significantly improved the efficacy of rosuvastatin in raising high-density lipoprotein cholesterol (HDL-C) [MD=2.38, 95%CI(0.61,4.16), P=0.009 0]. In the homozygous gene model, compared with TT genotype, CC genotype significantly improved the efficacy of rosuvastatin in reducing total cholesterol [MD=-7.50,95%CI(-13.05, -1.95), P=0.008 0]. In heterozygous gene model, compared with TT genotype, TC genotype significantly improved rosuvastatin in reducing low-density lipoprotein cholesterol (LDL-C) [MD=-5.14, 95%CI(-9.74, -0.53), P=0.03] and increasing HDL-C [MD=5.67, 95%CI 232102311200) (2.61, 8.73), P=0.000 3]. 388A>G gene polymorphism was also significantly correlated with the efficacy of rosuvastatin. In dominant or homozygous gene models, compared with AA E-mail:dushuzhang911@163.com genotype, GG+AG genotype [MD=-6.88, 95%CI (-7.46,-6.30),P<0.000 1] or GG genotype [MD=-9.23, 95%CI(-9.41, 9.04), P<0.000 1] significantly improved the efficacy of rosuvastatin in lowering LDL-C. In the heterozygous gene model, compared with AA genotype, AG genotype significantly improved the efficacy of rosuvastatin in lowering LDL-C [MD=-3.00, 95%CI(-3.19, -2.82), P<0.000 1], total cholesterol [MD=-5.80, 95%CI(-6.00, -5.59), P<0.000 1] and triglyceride [MD=-11.79, 95%CI(-19.57, -4.02), P=0.003 0]. In the recessive gene model, compared with AA+AG genotype, GG genotype significantly improved the therapeutic efficacy of rosuvastatin in reducing LDL-C[MD=-4.31, 95%CI(-8.47, -0.14), P=0.040 0] and elevating HDL-C [MD=4.49, 95%CI (2.20, 6.77), P=0.000 1]. Under 4 gene models, there was a significant correlation between 521T>C gene polymorphism and rosuvastatin-related ADR probability (P<0.05), but no significant correlation was found in 388A>G gene polymorphism (P>0.05). CONCLUSIONS The polymorphism of 521T>C gene is significantly related to the efficacy and safety of rosuvastatin in lowering lipid, and the C allele may be one of the factors leading to the increase of rosuvastatin in lipid-lowering efficacy and ADR. 388A> G gene polymorphism is significantly associated with the lipid-lowering efficacy of rosuvastatin, but not with its safety.
6.Study on identification of medicinal plant Gentiana scabra Bge. in Liaoning province based on DNA barcode sequences
Jianing XUE ; Rong ZHAO ; Ying YU ; Mingze GAO ; Haibo YIN
International Journal of Traditional Chinese Medicine 2024;46(1):103-107
Objective:To identify and analyze the genuine medicinal plant Gentiana scabra Bge. from 9 regions in Liaoning Province using DNA barcode technology based on the base sequence of internal transcribed spacer. Methods:DNA was extracted from the medicinal parts of 26 Gentiana scabra Bge. samples by using DNA kit extraction method. The ITS sequence was amplified through polymerase chain reaction (PCR), and then two-way sequencing was carried out. Other sources and outgroup sequences of the medicinal plant Gentiana scabra Bge. were downloaded from Genbank. After the sequencing results were spliced by using SeqMan 7.1.0 software, MEGA 7.0 software was used to analyze and compare the data, and calculate the genetic distance of K2P (Kimura 2-parameter). The phylogenetic tree was established by Neighbor-Joining (NJ) method for analysis. Results:According to the results of NJ cluster tree, all Gentiana scabra Bge. samples from different sources were clustered into one large branch, and Gentiana scabra Franch. and Gentiana triflora Pall. were clustered into one branch respectively, with obvious differences; Gentiana scabra and Gentiana manshurica Kitag. were clustered into one branch, and the genetic relationship was relatively close. In combination with the variation site and genetic distance, the base sequences of Gentiana scabra and Gentianamanshurica were very similar, and the interspecific differences were very small. Except for the intraspecific variation of only one sample collected in Liaoning Province, the base sequences of the other samples were the same, and there was no difference between " Gentiana scabra Bge. in Qingyuan" and Gentiana scabra Bge. samples from other regions in Liaoning Province. Conclusion:The DNA barcode technology of ITS sequence can be used to differentiate and identify medicinal plant Gentiana scabra Bge. and its original plants from different sources with a high success rate.
7.Study on DNA barcoding of Atractylodes chinensis (DC.) Koidz. herbs from Liaoning Province based on rbcL sequences
Ying YU ; Rong ZHAO ; Mingze GAO ; Jianing XUE ; Haibo YIN
International Journal of Traditional Chinese Medicine 2024;46(2):249-253
Objective:To use rbcL sequences to identify the rhizomes of the Liaoning collection of Atractylodes chinensis (DC.) Koidz.; To provide a basis for ensuring the feasibility of cultivation of the native herb in Liaoning Province. Methods:A total of 30 rhizomes of Atractylodes chinensis (DC.) Koidz. were collected from 10 regions cultivated in Liaoning Province, and the total DNA was extracted. DNA barcodes were screened by PCR, and the rbcL sequences of the samples were amplified and sequenced, and the amplification and sequencing success rates were calculated. Sequence alignment was performed using MEGA 7.0 software; a systematic clustering tree was constructed using the neighbour-joining method. Results:The success rates of DNA extraction from the rhizomes of Atractylodes chinensis (DC.) Koidz. were all 93.3%, and the success rates of PCR amplification and sequencing were all 100%. Among the 30 samples of Atractylodes chinensis (DC.) Koidz. in Liaoning Province, two samples had intraspecific variation, and the rest of the base sequences of Atractylodes chinensis (DC.) Koidz. were identical. Atractylodes chinensis (DC.) Koidz. was closer to the herbs of the genus Cangzhu, a relative species of Asteraceae, and was genetically more distant from the rest of Asteraceae. The NJ tree could distinguish Atractylodes chinensis (DC.) Koidz. and its relatives. Conclusion:The quality of Atractylodes chinensis (DC.) Koidz. cultivars in Liaoning Province is basically similar, and the rbcL sequence can be used as a valid sequence fragment for the identification of Atractylodes chinensis (DC.) Koidz. DNA barcode.
8. 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
9.Feasibility of non-invasive prenatal testing for the detection of fetal chromosomal copy number variants
Ying XUE ; Longwei QIAO ; Quanze HE ; Chunhua ZHANG ; Xiaojuan WU ; Guodong ZHAO ; Ting WANG
Chinese Journal of Medical Genetics 2024;41(6):696-701
Objective:To explore the feasibility of non-invasive prenatal testing (NIPT) for detecting fetal chromosomal copy number variants (CNV).Methods:A retrospective analysis was carried out on NIPT positive samples in Suzhou Municipal Hospital from January 1, 2019 to December 31, 2021. The effect of NIPT on fetal CNV detection was assessed by comparison with the results of karyotype analysis and/or chromosomal microarray analysis (CMA).Results:Among the 525 NIPT positive samples, 146 were CNV cases, of which 84 were further verified by karyotyping and/or CMA, 29(34.5%) were true positive. Among them, 12 cases were pathogenic variants, 2 cases were likely pathogenic variants and 15 cases were variants of uncertain significance.Conclusion:NIPT could detect CNV with high accuracy, and to combine CNV detection and chromosomal aneuploidy detection has great significance to improve the prenatal and postnatal care.
10.Improvement of isolation and culture methods for primary chondrocytes of neonatal rats
Dandan YANG ; Jiaoyang CHEN ; Xinheng WANG ; Zetong ZHAO ; Ying PAN ; Baigong XUE ; Changzhao GAO
Journal of Jilin University(Medicine Edition) 2024;50(5):1438-1449
Objective:To discuss the improved methods for the isolation and culture of primary chondrocytes from the neonatal rats,and to establish an efficient and economical in vitro chondrocyte culture system.Methods:The primary chondrocytes were isolated from the joints of neonatal rats and divided into overnight digestion(OD)group and rapid digestion(RD)group for separation.The chondrocytes in OD group were digested overnight by type Ⅱ collagenase,while the chondrocytes in RD group were separated by the combination of pre-digestion with physical and chemical digestion methods.The chondrocytes were cultured in modified media containing 0%(blank group 1),1%,2%,4%,and 10%fetal bovine serum(FBS),0(blank group 2),0.1,0.2,0.4,0.8,1.0,and 2.0 g·L-1 vitamin C(VC),and 0(blank group 3),0.5,1.0,2.0,4.0,8.0,10.0 μg·L-1 poly(lactic-co-glycolic acid)(PLGA)nanoparticles.The media containing different concentrations of FBS,VC,and PLGA were mixed with Dulbecco's modified Eagle's medium/nutrient mixture F-12(DMEM/F12),and were divided into related groups based on the concentrations of ingredients.Cell counter was used to count the chondrocytes in various groups and the survival rates and diameters of the chondrocytes in various groups were detected;Toluidine blue staining was used to detect the morphology of the chondrocytes in various groups;CCK-8 method was used to detect the proliferative activities of the chondrocytes in various groups;cell adhesion assay was used to detect the adhesion rates of the chondrocytes in various groups;Hoechst/propidium iodide(PI)staining was used to detect the apoptosis of the chondrocytes in various groups;MTT assay was used to detect the proliferation activities of the chondrocytes in various groups after treated with modified media.The cells were divided into DMEM/F12+10%FBS group,DMEM/F12+1%FBS group,and DMEM/F12+1%FBS+0.4 g·L-1 VC+1 μg·L-1 PLGA group.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of sex-determining region Y-box 9(SOX9),collagen type Ⅱ alpha 1 chain(Col2A1),collagen type X alpha 1 chain(Col10A1),and matrix metallopeptidase 13(MMP13)mRNAs in the chondrocytes in various groups after treated with modified media;immunofluorescence staining was used to detect the expressions of type Ⅱ collagen(COL Ⅱ)and SOX9 in the chondrocytes in various groups after treated with modified media.Results:The survival rate of primary chondrocytes in OD group was lower than that in RD group,and the average cell diameter was larger than that in RD group.The primary chondrocytes in OD group were larger and spindle-shaped,and most cells exhibited pseudopodia;in RD group,the primary chondrocytes were smaller,mostly rhomboid in shape,with only a portion of the cells showing pseudopodia.The Toluidine blue staining results showed significant coloration in both groups,but the digestion time of the chondrocytes in RD group was shorter,and compared with OD group,the actual culture time of the chondrocytes was reduced by 9-13 h,and more immature morphology of the primary chondrocytes were observed.The proliferation activity of the primary chondrocytes in OD group was slow at 24 h of culture but increased at 48 h of culture,and the proliferation activity of the primary chondrocytes was significantly higher at 48 h of culture compared with 12 h of culture(P<0.01).Compared with 12 h of culture,the proliferation rates of the primary chondrocytes in RD group were increased at 24 and 48 h of culture(P<0.01).At 24 and 48 h of culture,compared with OD group,the proliferation rates of the primary chondrocytes in RD group were increased(P<0.05).The number of apoptotic chondrocytes in RD group was lower than that in OD group,and no necrotic chondrocytes were observed in either group.The proliferation activities of chondrocytes of the rats were increased with the rising of FBS concentration in the culture medium.Compared with blank group 1,the proliferation activities of chondrocytes of the rats after treated with culture mediums containing 1%,2%,4%,and 10%FBS were significantly increased(P<0.05).Compared with blank group 2,the proliferative activities of chondrocytes of the rats after treated with culture mediums containing 0.2-1.0 g·L-1 VC were significantly increased(P<0.05),and the highest proliferation activity was found when the concentration of VC was 0.4 g·L-1(P<0.01).Compared with blank group 3,the proliferation activities of chondrocytes of the rats after treated with culture mediums containing 1-4 μg·L-1 PLGA were significantly increased(P<0.05),and the highest proliferation activity was found after treated with culture medium containing 1 μg·L-1 PLGA(P<0.05).Compared with DMEM/F12+10%FBS group,the expression levels of SOX9 mRNA and Col2A1 mRNA in the chondrocytes in DMEM/F12+1%FBS group were significantly increased(P<0.05 or P<0.01).Compared with DMEM/F12+10%FBS group,the expression levels of SOX9 mRNA and Col2A1 mRNA in the chondrocytes in DMEM/F12+1%FBS+0.4 g·L-1 VC+1 μg·L-1 PLGA group were significantly increased(P<0.01).The immunofluorescence staining results showed that the green fluorescence signal of COL Ⅱ and the red fluorescence signal of SOX9 were observed in some chondrocytes in DMEM/F12+10%FBS group under fluorescence microscope,and the fluorescence intensity was weak.In DMEM/F12+1%FBS group,most chondrocytes exhibited COL Ⅱ green fluorescence signal and SOX9 red fluorescence signal,and the fluorescence intensity was significantly stronger than that in DMEM/F12+10%FBS group.In DMEM/F12+1%FBS+0.4 g·L-1 VC+1 μg·L-1 PLGA group,the COLⅡ green fluorescence signal and SOX9 red fluorescence signal were found in all the chondrocytes,and the fluorescence intensity was significantly higher than those in DMEM/F12+10%FBS and DMEM/F12+1%FBS groups.The expression levels of COLⅡ and SOX9 proteins in the chondrocytes in DMEM/F12+1%FBS group were significantly higher than those in DMEM/F12+10%FBS group,and the expression levels of COL Ⅱ and SOX9 proteins in the chondrocytes in DMEM/F12+1%FBS+0.4 g·L-1 VC+1 μg·L-1 PLGA group were significantly higher than those in DMEM/F12+10%FBS group.Conclusion:The improved methods for the isolation and culture of primary chondrocytes of the rats can overcome the shortcomings of traditional methods,shorten the isolation time of primary chondrocytes,and improve the quality of in vitro culture of primary chondrocytes.

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