1.Updates and amendments of the Chinese Pharmacopoeia 2025 Edition (Volume Ⅰ)
LI Hao ; SHEN Mingrui ; ZHANG Pang ; ZHAI Weimin ; NI Long ; HAO Bo ; ZHAO Yuxin ; HE Yi ; MA Shuangcheng ; SHU Rong
Drug Standards of China 2025;26(1):017-022
The Chinese Pharmacopoeia is the legal technical standard which should be followed during the research, production, use, and administration of drugs. At present, the new edition of the Chinese Pharmacopoeia is planned to be promulgated and implemented. This article summarizes and analyzes the main characteristics and the content of updates and amendments of the Chinese Pharmacopoeia 2025 Edition(Volume Ⅰ), to provide a reference for the correct understanding and accurate implementation the new edition of the pharmacopoeia.
2.Clinical Observation and Mechanism Study of Xu's Shenqi Yizhu Decoction Combined with Chemotherapy in the Treatment of Cancer-related Fatigue of Stagnated-toxin Spleen Deficiency Type in Postoperative Gastric Cancer Patients
Zhixiang SHEN ; Weimin LU ; Chuan SHI ; Mengwei YE ; Mengyuan LEI ; Ziyun LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):143-151
ObjectiveTo evaluate the clinical efficacy of Xu's Shenqiyizhu (SQYZ) decoction combined with chemotherapy in the treatment of cancer-related fatigue (CRF) of stagnated-toxin spleen deficiency type after gastric cancer surgery and explore its possible mechanism. MethodsFifty postoperative gastric cancer patients with CRF of stagnated-toxin spleen deficiency type were selected and randomly divided into an experimental group and a control group by using a random number table,with 25 cases in each group. The control group was treated with FLOT chemotherapy (50 mg·m-2 docetaxel (iv drip on day 1) + 85 mg·m-2 oxaliplatin (iv drip on day 1) + 200 mg·m-2 calcium folinate (iv drip on day 1) + 2 600 mg·m-2 fluorouracil (iv drip for 24 h on day 1),once every three weeks) and basic and symptomatic supportive treatment. The experimental group was treated with Xu's SQYZ decoction (decocted twice,200 mL taken orally twice a day) in addition to the treatment of the control group. One course of treatment lasted for three weeks,with a total of four courses conducted. Observation was performed on the piper fatigue scale (PFS) scores,karnofsky performance status (KPS) scores,European Organization for Research and Treatment of cancer quality of life questionnaire (EORTC QLQ-C30) scores,traditional Chinese medicine syndrome scores,and serum levels of tumor necrosis factor-α (TNF-α),interferon-γ (IFN-γ),and interleukin-6 (IL-6)detected via enzyme linked immunosorbent assay (ELISA) before and after treatment in the two groups. The safety test results before and after treatment for the two groups of patients,as well as the occurrence of adverse events during treatment, were recorded. Transcriptome sequencing data of peripheral blood samples from gastric adenocarcinoma patients and normal individuals were downloaded from the gene expression omnibus (GEO) database,and differentially expressed genes between the tumor and normal groups were identified. Differential gene enrichment analysis was made based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO). The CRF relevance scores of genes were retrieved from the GeneCards database. Results① Compared with that before treatment,the total PFS score in the experimental group was significantly reduced (P<0.05). Compared with the control group after treatment,the experimental group showed significantly reduced total PFS score (P<0.05). ② Compared with that before treatment,the KPS score in the experimental group decreased significantly (P<0.05). Compared with the control group after treatment,the experimental group exhibited a significantly decreased KPS score (P<0.05). The experimental group demonstrated significantly increased functional scores (physical function,role function,emotional function,social function,and overall health) (P<0.05) and significantly reduced symptom scores (fatigue,shortness of breath,loss of appetite,constipation,and diarrhea) of the EORTC QLQ-C30 scale after treatment compared with before treatment. Compared with the control group after treatment,the experimental group presented significantly increased functional scores (physical function,emotional function,social function,and overall health) (P<0.05) and significantly reduced symptom scores (fatigue,nausea and vomiting,shortness of breath,loss of appetite,and diarrhea) of the EORTC QLQ-C30 scale (P<0.05). Compared with those before treatment,the traditional Chinese medicine syndrome scores (eating too little and poor digestion,fatigue and weakness,postprandial bloating,abnormal bowel movements,lassitude and weakness,and total score) in the experimental group were significantly reduced (P<0.05). Compared with the control group after treatment,the experimental group had significantly reduced traditional Chinese medicine syndrome scores (eating too little and poor digestion,fatigue and weakness,nausea and vomiting,and sallow complexion) (P<0.05), which indicated better efficacy in the experimental group than in the control group (χ2=7.996,P<0.05). The serum levels of TNF-α,IL-6,and IFN-γ were significantly correlated with each other (P<0.01). Compared with those before treatment,the levels of serum cytokines TNF-α,IL-6,and IFN-γ in the experimental group were significantly reduced (P<0.05). Compared with the control group after treatment,the experimental group showed significantly reduced serum levels of cytokines TNF-α,IL-6,and IFN-γ (P<0.05). ③ There were no significant intra-group and inter-group differences in the safety test results of the two groups before and after treatment. During the treatment period,there was no significant difference in the incidence of adverse reactions between the two groups of patients. ④ Compared with the normal group,the tumor group exhibited a total of 328 significantly up-regulated genes in the peripheral blood (P<0.05),and KEGG and GO analyses showed that they were significantly enriched in signaling pathways such as TNF (P<0.05). ⑤ TNF,IL6,IFNG, and other cytokine encoding genes may be key pathogenic genes for CRF. ConclusionXu's SQYZ decoction can alleviate symptoms such as fatigue in postoperative chemotherapy patients with gastric cancer and improve their functional status and quality of life. Its mechanism may be related to improving cytokine imbalance.
3.Fronto-orbital plasty using premature frontal suture osteotomy with frontoparietal transposition for the treatment of metopic synostosis
Jun YAN ; Jie CUI ; Yi JI ; Weimin SHEN
Chinese Journal of Plastic Surgery 2024;40(10):1059-1064
Objective:To evaluate the efficacy and safety of fronto-orbital plasty using premature frontal suture osteotomy with frontoparietal transposition for the treatment of metopic synostosis.Methods:Retrospective analysis was performed on the clinical data of children with metopic synostosis who were treated in the Department of Burns and Plastic Surgery, Children’ s Hospital of Nanjing Medical University from January 2015 to October 2023 with fronto-orbital plasty, which involved premature frontal suture osteotomy with frontoparietal transposition. The frontal and parietal bones were divided into two bones, then the supraorbital bridge was cut off and the premature frontal suture was removed. The parietal bone block was then placed on the upper edge of the orbital bone, followed by the placement of the two pieces of the frontal bone on the upper edge of the parietal bone. Finally, all bone blocks were secured using absorbable plates. Pre-and post-operative CT scans and reconstructions were performed to measure changes in cranial cavity volume, and complications such as cerebrospinal fluid leakage, infection, and recurrence of metopic synostosis were followed up.Results:A total of 5 patients with metopic synostosis were enrolled, 4 males and 1 female, aged 5 to 14 months. The operation time was 4.5 to 5.5 hours, and the intraoperative blood loss was 50 to 150 ml. All patients completed the treatment without serious complications. The cranial cavity volume increased by 48.53 to 172.00 cm 3 after the operation. Follow-up was conducted for 3 to 6 months, and the trigonocephaly was completely improved without any serious complications. Short-term follow-up indicated stable craniofacial morphology. Conclusion:Fronto-orbital plasty using premature frontal suture osteotomy with frontoparietal transposition is an effective and safe treatment for metopic synostosis. The procedure has a low risk of recurrence and complications, and the result are positive. Recurrence was scarce in the short term, and the long-term situation needs to be observed.
4.Application effect of allogeneic acellular dermal matrix in the repair of nail fold asymmetry deformity after complete syndactyly reconstruction in children
Zhengfu YU ; Jie CUI ; Weimin SHEN ; Jijun ZOU
Chinese Journal of Plastic Surgery 2024;40(11):1168-1174
Objective:To explore the clinical application effect of allogeneic acellular dermal matrix (ADM) in the repair of asymmetric nail fold deformity after complete syndactyly surgery in children.Methods:A retrospective analysis was conducted on the clinical data of children with secondary asymmetric nail fold deformity after complete syndactyly surgery treated at the Department of Burn and Plastic Surgery, Children’s Hospital of Nanjing Medical University, from January 2022 to December 2023. The patients required a secondary surgical repair due to insufficient soft tissue capacity with or without scar contracture, causing asymmetry of the nail fold after finger separation at the distal end following syndactyly surgery. During the operation, continuous Z-plasty was designed along the original surgical scar axis and incised towards the proximal end of the lateral nail fold, followed by thorough scar release. Scar debulking surgery was performed if there was significant local scar hyperplasia. A tunnel was created from the incision site near the lateral nail fold to reach the fingertip subcutaneously. ADM was filled into this tunnel until a satisfactory appearance of the lateral nail fold was achieved. The flaps were then closed by cross-suturing. Regular follow-up observations were conducted on the incision healing, scar, and appearance of the finger, and aesthetic evaluation according to Bulic’s standards was performed, classifying them into four grades: excellent, good, fair, and poor.Results:The study enrolled 12 patients, and nail fold asymmetry appeared 3 to 6 months after surgery, with 8 males and 4 females, ranging from 1 to 9 years old (average age 4.6 years). Affected digits included both hands in 7 cases and one hand in 5 cases, totaling 38 fingers. All 12 cases including 25 fingers presented with insufficient distal capacity in their fingers; seven cases including 16 fingers also had scar contracture resulting in poor appearance. All procedures were successfully performed. Post-operatively, one case of necrosis at the tip of the scar flap was observed after 12 days of removing the dressing but healed after dressing changes, and no infections or flap necrosis occurred in the remaining patients, with the incisions healing primarily. Follow-up periods ranged from 1 to 24 months, with an average duration of 13.6 months. All patients achieved the restoration of soft tissue capacity in the finger lateral nail fold, varying degrees of correction for asymmetrical nail fold deformities, and complete release for scar contracture, with 7 fingers in 4 cases rated as excellent, 12 fingers in 5 cases as good, 4 fingers in 2 cases as fair, and 2 fingers in one case as poor in evaluation of fingertip appearance.Conclusion:The use of ADM for filling is effective in restoring soft tissue capacity and correcting asymmetrical deformities caused by secondary nail fold abnormalities after complete syndactyly repair surgery in children. It has shown good clinical outcomes with minimal complications.
5.Fronto-orbital plasty using premature frontal suture osteotomy with frontoparietal transposition for the treatment of metopic synostosis
Jun YAN ; Jie CUI ; Yi JI ; Weimin SHEN
Chinese Journal of Plastic Surgery 2024;40(10):1059-1064
Objective:To evaluate the efficacy and safety of fronto-orbital plasty using premature frontal suture osteotomy with frontoparietal transposition for the treatment of metopic synostosis.Methods:Retrospective analysis was performed on the clinical data of children with metopic synostosis who were treated in the Department of Burns and Plastic Surgery, Children’ s Hospital of Nanjing Medical University from January 2015 to October 2023 with fronto-orbital plasty, which involved premature frontal suture osteotomy with frontoparietal transposition. The frontal and parietal bones were divided into two bones, then the supraorbital bridge was cut off and the premature frontal suture was removed. The parietal bone block was then placed on the upper edge of the orbital bone, followed by the placement of the two pieces of the frontal bone on the upper edge of the parietal bone. Finally, all bone blocks were secured using absorbable plates. Pre-and post-operative CT scans and reconstructions were performed to measure changes in cranial cavity volume, and complications such as cerebrospinal fluid leakage, infection, and recurrence of metopic synostosis were followed up.Results:A total of 5 patients with metopic synostosis were enrolled, 4 males and 1 female, aged 5 to 14 months. The operation time was 4.5 to 5.5 hours, and the intraoperative blood loss was 50 to 150 ml. All patients completed the treatment without serious complications. The cranial cavity volume increased by 48.53 to 172.00 cm 3 after the operation. Follow-up was conducted for 3 to 6 months, and the trigonocephaly was completely improved without any serious complications. Short-term follow-up indicated stable craniofacial morphology. Conclusion:Fronto-orbital plasty using premature frontal suture osteotomy with frontoparietal transposition is an effective and safe treatment for metopic synostosis. The procedure has a low risk of recurrence and complications, and the result are positive. Recurrence was scarce in the short term, and the long-term situation needs to be observed.
6.Application effect of allogeneic acellular dermal matrix in the repair of nail fold asymmetry deformity after complete syndactyly reconstruction in children
Zhengfu YU ; Jie CUI ; Weimin SHEN ; Jijun ZOU
Chinese Journal of Plastic Surgery 2024;40(11):1168-1174
Objective:To explore the clinical application effect of allogeneic acellular dermal matrix (ADM) in the repair of asymmetric nail fold deformity after complete syndactyly surgery in children.Methods:A retrospective analysis was conducted on the clinical data of children with secondary asymmetric nail fold deformity after complete syndactyly surgery treated at the Department of Burn and Plastic Surgery, Children’s Hospital of Nanjing Medical University, from January 2022 to December 2023. The patients required a secondary surgical repair due to insufficient soft tissue capacity with or without scar contracture, causing asymmetry of the nail fold after finger separation at the distal end following syndactyly surgery. During the operation, continuous Z-plasty was designed along the original surgical scar axis and incised towards the proximal end of the lateral nail fold, followed by thorough scar release. Scar debulking surgery was performed if there was significant local scar hyperplasia. A tunnel was created from the incision site near the lateral nail fold to reach the fingertip subcutaneously. ADM was filled into this tunnel until a satisfactory appearance of the lateral nail fold was achieved. The flaps were then closed by cross-suturing. Regular follow-up observations were conducted on the incision healing, scar, and appearance of the finger, and aesthetic evaluation according to Bulic’s standards was performed, classifying them into four grades: excellent, good, fair, and poor.Results:The study enrolled 12 patients, and nail fold asymmetry appeared 3 to 6 months after surgery, with 8 males and 4 females, ranging from 1 to 9 years old (average age 4.6 years). Affected digits included both hands in 7 cases and one hand in 5 cases, totaling 38 fingers. All 12 cases including 25 fingers presented with insufficient distal capacity in their fingers; seven cases including 16 fingers also had scar contracture resulting in poor appearance. All procedures were successfully performed. Post-operatively, one case of necrosis at the tip of the scar flap was observed after 12 days of removing the dressing but healed after dressing changes, and no infections or flap necrosis occurred in the remaining patients, with the incisions healing primarily. Follow-up periods ranged from 1 to 24 months, with an average duration of 13.6 months. All patients achieved the restoration of soft tissue capacity in the finger lateral nail fold, varying degrees of correction for asymmetrical nail fold deformities, and complete release for scar contracture, with 7 fingers in 4 cases rated as excellent, 12 fingers in 5 cases as good, 4 fingers in 2 cases as fair, and 2 fingers in one case as poor in evaluation of fingertip appearance.Conclusion:The use of ADM for filling is effective in restoring soft tissue capacity and correcting asymmetrical deformities caused by secondary nail fold abnormalities after complete syndactyly repair surgery in children. It has shown good clinical outcomes with minimal complications.
7.The effect of preoperative patellar tilt angle on postoperative outcome of total knee arthroplasty
Hao ZHOU ; Kai SHEN ; Jinwen WANG ; Zhenggang TAN ; Weimin FAN ; Feng LIU ; Jiuxiang LIU
Chinese Journal of Orthopaedics 2024;44(9):594-600
Objective:To investigate the effect of preoperative patellar tilt angle on postoperative outcome of total knee arthroplasty (TKA).Methods:A total of 277 patients with knee osteoarthritis who underwent TKA without patellar replacement in the First Affiliated Hospital of Nanjing Medical University from October 2020 to September 2022 were retrospectively analyzed. There were 72 males and 205 females, aged 69.16±6.77 years (range, 52-87 years), body mass index 27.01±3.81 kg/m 2 (range 18.14-39.01 kg/m 2). The patients were divided into three groups according to the preoperative tilt angle of the patella: tilt angle<5° for mild tilt group, 5°≤tilt angle<10° for moderate tilt group, and tilt angle≥10° for severe tilt group. There were 103 cases in the mild group, 137 cases in the moderate group, and 37 cases in the severe group. The preoperative and postoperative visual analogue scale (VAS), Hospital for Special Surgery (HSS) score and Feller's score of patella were compared. Results:All patients were followed up for 15.73±3.06 months (range, 12-22 months). The patellar tilt angle was 6.86°±3.55° preoperatively and 3.63°±2.61° postoperatively ( t=19.086, P<0.001). The patellar tilt angle of mild group, moderate group and severe group decreased after operation, and the difference was statistically significant compared with that before operation ( P<0.05). The anterior knee VAS of the three groups decreased after operation, and the differences were statistically significant compared with those before operation. The anterior knee VAS at 3, 6, and 12 months after operation were lower than those before operation. The differences between groups at 6 and 12 months postoperatively were statistically significant ( P<0.05), with anterior knee VAS scores of 2.59±0.55 and 2.03±0.55 in the severe group being greater than those of 2.15±0.38 and 1.57±0.50 in the mild group and 2.19±0.49 and 1.67± 0.61 in the moderate group. The HSS score of the three groups was increased after operation, and the difference was statistically significant compared with that before operation ( P<0.05), and the HSS scores at 3, 6 and 12 months after operation were higher than those before operation. There were significant differences at 6 and 12 months after operation ( P<0.05), the HSS scores of the severe group were 86.27±2.04 and 87.73±2.28, which were lower than those of the mild group 89.02±2.33 and 89.83±1.48, and the moderate group 88.77±2.83 and 89.52±1.95. Postoperative patellar Feller score increased in all three groups, and the difference was statistically significant compared with that before operation ( P<0.05), the patellar Feller score at 3, 6 and 12 months after operation was higher than that before operation. There were significant differences at 6 and 12 months after operation ( P<0.05), the patellar Feller scores in the severe group were 18.32±1.99 and 20.32±1.60, which were lower than those in the mild group 20.92±1.01 and 23.07±1.39 and the moderate group 20.91±1.95 and 22.69±1.59. Conclusion:In TKA patients without patella replacement, the increase of patellar tilt angle before operation can lead to anterior knee pain and reduced knee function.
8.Mechanism of baicalein in alleviating osteoarthritis by inhibiting fibroblast-like synoviocytes senescence through Sirt1
Yong XIAO ; Kai SHEN ; Hao ZHOU ; Chenjun ZHAI ; Jinchun ZHOU ; Feng LIU ; Weimin FAN
Chinese Journal of Orthopaedics 2024;44(13):889-899
Objective:To investigate the mechanism by which baicalein alleviates osteoarthritis by regulating Sirtuin (Sirt) to inhibit the senescence of fibroblast-like synoviocytes (FLS).Methods:FLS were extracted from synovium of non-osteoarthritis patients (3 patients) and osteoarthritis patients (3 patients), and assessed the senescence of FLS in osteoarthritis patients by Western blot, β-galactosidase staining and immunofluorescence, and examined the changing levels of Sirt family proteins and mRNAs in FLS. The relative expression level of Sirt1 in FLS was knocked down by transfection with Sirt1 siRNA, and the antioxidative capacity and senescence degree of FLS were analyzed. Chondrocytes were co-cultured with FLS and functional changes in chondrocytes were detected by Western blot. After inducing FLS senescence with bleomycin (BLM), different concentrations of baicalein were administered to detect the expression levels of Sirt1, p16, and p21 proteins and intracellular reactive oxygen species (ROS). The expression level of Sirt1 in FLS was knocked down, and BLM and baicalein treatments were applied. The relative expression levels of Sirt1 and ROS were detected using Western blot and ROS assays. After BLM-induced FLS senescence, baicalein and Compound C were added. The relative expression levels of phosphorylated AMP-activated protein kinase (pAMPK), nuclear factor erythroid 2-related factor 2 (NRF2), p16, p21, and ROS were detected by Western blot and ROS assays.Results:The relative expression of p16 and p21 in FLS in the osteoarthritic group were 3.66±0.38 and 3.55±0.34, which were higher than those in non-osteoarthritis group 1.00±0.07 and 1.00±0.09 ( t=11.860, P<0.001; t=12.520, P<0.001). The relative expression levels of Sirt1 and Sirt6 in FLS in the osteoarthritis group were 0.30±0.04 and 0.16±0.01, which were smaller than those in the non-osteoarthritis group 1.00 ± 0.03 and 1.00±0.04 ( t=23.840, P<0.001; t=34.130, P<0.001). After baicalein treatment, the relative expression of ROS was 2.58±0.28, 1.65±0.14 and 1.00±0.24 in the BLM, BLM+Bai and control groups, respectively, with statistically significant differences between the groups ( F=35.700, P<0.001), which was greater in the BLM and BLM+Bai than in the control group, and lower in the BLM+Bai than in the BLM group ( P<0.05). The relative expression of pAMPK and NRF2 was 0.28±0.02 and 0.38±0.09 after Compound C+Baicalein treatment, which was lower than that of 0.56±0.07 and 0.60±0.08 in the BLM+Bai group ( P<0.05). The relative expression of ROS increased from 1.75±0.16 to 3.45±0.12 ( P<0.001). The proportion of positivity in the BLM+Bai+Compound C, BLM+Bai and control groups was 47.30%±4.29%, 18.18%±3.89% and 7.70%±3.53% ( F=109.700, P<0.001), respectively, with the BLM+Bai+Compound C group being higher than that in the BLM+Bai group ( P<0.05). Conclusion:The downregulation of Sirt1 expression in patients with osteoarthritis leads to FLS senescence and accelerates the progression of osteoarthritis. Baicalein can inhibit FLS senescence by activating Sirt1/AMPK/NRF2 pathway, which may delay the progression of osteoarthritis and improve the function of chondrocytes.
9.Expression of miR-15b and miR-503 in peripheral blood and their relationship with cognitive function in patients with depression
Weimin HE ; Xiaofeng JIANG ; Qundi SHEN ; Ying CHEN
China Modern Doctor 2024;62(27):38-42
Objective To investigate the expression of microRNA(miR)-15b and miR-503 in peripheral blood in patients with depression and their relationship with cognitive function.Methods A total of 103 patients with depression admitted and diagnosed in Shaoxing Seventh People's Hospital from January 2023 to June 2024 were set as study group,and 94 healthy participants were selected as control group.The cognitive function of patients with depression was evaluated by Montreal cognitive assessment(MoCA),and the patients in study group were divided into normal cognitive function group(n=68)and cognitive dysfunction group(n=35).Pearson method was used to analyze the correlation between the expression level of miR-15b,miR-503 and cognitive function;The diagnostic value of miR-15b and miR-503 for cognitive dysfunction in patients with depression was analyzed by using receiver operating characteristic(ROC)curve.Results miR-15b and miR-503 in the peripheral blood of study group were lower than those of control group(P<0.05),miR-15b and miR-503 in the peripheral blood of cognitive dysfunction group were lower than those of normal cognitive function group(P<0.05),the scores of MoCA and mini-mental state examination in normal cognitive function group were higher than those in cognitive dysfunction group(P<0.05);The results of Pearson analysis showed that the levels of miR-15b and miR-503 in peripheral blood of patients with depression were significantly positively correlated with MoCA scores(r=0.635,0.668,P<0.05);The area under the curve(AUC)of miR-15b level alone in diagnosing cognitive dysfunction in patients with depression was 0.836(95%CI:0.750-0.902),the sensitivity and specificity were 77.14%and 89.71%respectively;The AUC of miR-503 level alone in diagnosing cognitive dysfunction in patients with depression was 0.892(95%CI:0.816-0.945),the sensitivity and specificity were 80.00%and 88.24%respectively;The AUC for the combined diagnosis of miR-15b and miR-503 was significantly greater than the AUC for miR-15b and miR-503 alone.Conclusion The expression of miR-15b and miR-503 is decreased in patients with depression,and they are of high diagnostic value for cognitive dysfunction in patients with depression.
10.Exosomes secreted by hypoxia-preconditioned BMSCs promote cartilage regeneration
Kai SHEN ; Chenjun ZHAI ; Qiang ZUO ; Wenwei LIANG ; Weimin FAN ; Feng LIU
Chinese Journal of Orthopaedics 2023;43(12):831-840
Objective:Observing the effect of exosomes derived from hypoxic Bone marrow mesenchymal stem cells (BMSCs) on the function of chondrocytes, and exploring the role and mechanism of exosomal miR-196b-5p. Evaluating the application prospects of hypoxic BMSCs exosomes and miR-196b-5p for cartilage regeneration.Methods:Chondrocytes were cultured in the supernatant of BMSCs cultured under normoxia or hypoxia, respectively. The proliferation of chondrocytes was detected by CCK-8 assay and the expressions of Collagen type 2 (Col2), Col1, Aggrecan and SOX9 were detected by qPCR to evaluate the effect of hypoxic BMSCs paracrine on chondrocyte functions. Obtaining normoxic and hypoxic exosomes through ultracentrifugation, and testing their effects on the proliferation and anabolic-related genes of chondrocytes through CCK-8 assay and qPCR. Verifying the expression of miR-196b-5p in hypoxic exosomes based on exosomal miRNA array. Knocking out miR-196b-5p in hypoxic BMSCs, and detecting the effect of hypoxic exosomal miR-196b-5p on the functions of chondrocytes by loss-of-function assay. Predicting the downstream of miR-196b-5p through bioinformatics tools, and exploring the mechanism of hypoxic exosomal miR-196b-5p by gain-of-function assays. Hypoxic exosomes and miR-196b-5p-knockout hypoxic exosomes were loaded on silk fibroin hydrogel and subcutaneously into nude mice. After 4 weeks of culture, histological staining of saffron O, Masson and biochemical content of sGAG and collagen were performed to assess the application prospect of hypoxic exosomes and hypoxic exosomal miR-196b-5p on cartilage regeneration. Results:The results of CCK-8 assay and qPCR indicated that the supernatant of hypoxic BMSCs significantly promoted the proliferation of chondrocytes 1.20±0.07 and the expression of cartilage-related markers (Col2 2.95±0.17, Aggrecan 2.45±0.27, SOX9 2.92±0.29) compared to normoxic BMSCs (0.94±0.04, 1.89±0.09, 1.67±0.21, 1.76±0.16), the differences were statistically significant ( P<0.05). The result of CCK-8 assay showed that hypoxic exosomes (1.28±0.04) promoted the proliferation of chondrocytes compared to normoxic exosomes 1.05±0.06, the differences were statistically significant ( P<0.05). CCK-8 assay revealed that the down-regulation of miR-196b-5p in hypoxic exosomes 0.99±0.06 attenuated the proliferation of chondrocytes compared to control group 1.20±0.07, the differences were statistically significant ( P<0.05); the expression of Col2 0.56±0.04, Aggrecan 0.74±0.09, and SOX9 0.45±0.05 in chondrocytes was reduced in the miR-196b-5p knockdown group compared to the control group (1.00±0.09, 1.00±0.12, 1.00±0.07), the differences were statistically significant ( P<0.05). Co-transfection of pmirGLO-BACH1-WT reporter vector with miR-196b-5p mimics decreased the luciferase activity 0.73±0.06, the differences were statistically significant ( P<0.05). Co-transfection of pmirGLO-BACH1-MUT reporter vector with miR-196b-5p mimics showed no change in luciferase activity. BACH1 is the target of miR-196b-5p. Subcutaneous culture in nude mice showed that hypoxic exosomes significantly promoted the deposition of sGAG 383.2±21.54 and collagen 67.40±3.45, while reducing the expression of miR-196b-5p in hypoxic exosomes weakened the deposition of sGAG 258.4±19.50 and collagen 57.15±4.95, the differences were statistically significant ( P<0.05). Conclusion:Hypoxic exosomes promoted the functions of chondrocytes by inhibiting the expression of BACH1 through miR-196b-5p. Hypoxic exosomes can be applied in cartilage regeneration.

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