1.Whole genome sequencing of Bifidobacterium animalis subsp. lactic B4 and analysis of its probiotic properties
Ruipei Ding ; Chengzhong Liu ; Cancan Shi ; Huiyu Zhang ; Yiping Dai ; Ruxue Mu ; Zilong Li ; Maozhang He ; Zhuojun Li
Acta Universitatis Medicinalis Anhui 2025;60(2):255-265
Objective :
To investigate whole genome information of a newly isolatedBifidobacterium animalissubsp. lactic B4 strain from healthy human feces was analyzed and its probiotic properties.
Methods :
The antimicrobial resistance, hemolytic, gastric acid tolerance and biochemical characteristics of B. animalis B4 were evaluated byin vitroexperiments, and its whole genome was sequentially sequenced and functional annotation was performed by next and three-generation sequencing technology.
Results:
Whole genome sequencing of B. animalis B4 showed that its genome size was 1 944 146 bp, with GC content of 60.49%, no plasmid, and a total of 1 642 genes. The results ofin vitroanalysis showed that the B. animalis B4 had good probiotic properties, including non-hemolytic and stomach acid resistance. At the same time, the genome results showed that the B. animalis B4 strain did not have toxin and disease-related genes, drug resistance genes were few and the transmission ability was not high, so it had high safety. Gene annotation of KEGG, COG and GO showed that it contained many biological active enzymes, such as β-galactosidase, L-lactate dehydrogenase and other probiotic genes.
Conclusion
The B. animalis B4 has good probiotic properties, showing excellent safety at the genetic level, with a probiotic gene sequence.
2.Qingshen Granules alleviates renal fibrosis in mice by regulating exosomes,miR-330-3p,and CREBBP expression
Rong DAI ; Zeping CAO ; Chuanjiao LIU ; Yong GE ; Meng CHENG ; Weili WANG ; Yizhen CHEN ; Lei ZHANG ; Yiping WANG
Journal of Southern Medical University 2024;44(8):1431-1440
Objective To explore the effects of Qingshen Granules(QSG)on adenine-induced renal fibrosis in mice and in uric acid(UA)-stimulated NRK-49F cells and its mechanism for regulating exosomes,miR-330-3p and CREBBP.Methods A mouse model of adenine-induced renal fibrosis were treated daily with QSG at 8.0 g·kg-1·d-1 via gavage for 12 weeks.An adeno-associated virus vector was injected into the tail vein,and renal tissues of the mice were collected for analyzing exosomal marker proteins CD9,Hsp70,and TSG101 and expressions of Col-Ⅲ,α-SMA,FN,and E-cad using Western blotting and immunofluorescence and for observing pathological changes using HE and Masson staining.In the cell experiment,NRK-49F cells were stimulated with uric acid(400 μmol/L)followed by treatment with QSG-medicated serum from SD rats,and the changes in expressions of the exosomal markers and Col-Ⅲ,α-SMA,FN,and E-cad were analyzed.Dual luciferase reporter assay was employed to examine the targeting relationship between miR-330-3p and CREBBP,whose expressions were detected by RT-qPCR and Western blotting in treated NRK-49F cells.Results The mouse models of adenine-induced renal fibrosis showed significantly increased levels of CD9,Hsp70,and TSG101,which were decreased by treatment with QSG.The expressions of Col-Ⅲ,α-SMA,and FN increased and E-cad decreased in the mouse models but these changes were reversed by QSG treatment.QSG treatment obviously alleviated renal fibrosis in the mouse models.Intravenous injection of adeno-associated viral vector obviously inhibited miR-330-3p,increased CREBBP levels,and reduced fibrosis in the mouse models.Dual luciferase assay confirmed CREBBP as a target of miR-330-3p,which was consistent with the results of the cell experiments.Conclusion QSG inhibits renal fibrosis in mice by regulating the exosomes,reducing miR-330-3p levels,and increasing CREBBP expression.
3.Qingshen Granules alleviates renal fibrosis in mice by regulating exosomes,miR-330-3p,and CREBBP expression
Rong DAI ; Zeping CAO ; Chuanjiao LIU ; Yong GE ; Meng CHENG ; Weili WANG ; Yizhen CHEN ; Lei ZHANG ; Yiping WANG
Journal of Southern Medical University 2024;44(8):1431-1440
Objective To explore the effects of Qingshen Granules(QSG)on adenine-induced renal fibrosis in mice and in uric acid(UA)-stimulated NRK-49F cells and its mechanism for regulating exosomes,miR-330-3p and CREBBP.Methods A mouse model of adenine-induced renal fibrosis were treated daily with QSG at 8.0 g·kg-1·d-1 via gavage for 12 weeks.An adeno-associated virus vector was injected into the tail vein,and renal tissues of the mice were collected for analyzing exosomal marker proteins CD9,Hsp70,and TSG101 and expressions of Col-Ⅲ,α-SMA,FN,and E-cad using Western blotting and immunofluorescence and for observing pathological changes using HE and Masson staining.In the cell experiment,NRK-49F cells were stimulated with uric acid(400 μmol/L)followed by treatment with QSG-medicated serum from SD rats,and the changes in expressions of the exosomal markers and Col-Ⅲ,α-SMA,FN,and E-cad were analyzed.Dual luciferase reporter assay was employed to examine the targeting relationship between miR-330-3p and CREBBP,whose expressions were detected by RT-qPCR and Western blotting in treated NRK-49F cells.Results The mouse models of adenine-induced renal fibrosis showed significantly increased levels of CD9,Hsp70,and TSG101,which were decreased by treatment with QSG.The expressions of Col-Ⅲ,α-SMA,and FN increased and E-cad decreased in the mouse models but these changes were reversed by QSG treatment.QSG treatment obviously alleviated renal fibrosis in the mouse models.Intravenous injection of adeno-associated viral vector obviously inhibited miR-330-3p,increased CREBBP levels,and reduced fibrosis in the mouse models.Dual luciferase assay confirmed CREBBP as a target of miR-330-3p,which was consistent with the results of the cell experiments.Conclusion QSG inhibits renal fibrosis in mice by regulating the exosomes,reducing miR-330-3p levels,and increasing CREBBP expression.
4.Rotator cuff repair combined with transposition of the long head of the biceps tendon for massive rotator cuff tears with or without subscapularis injury
Jinsong YANG ; Zhihui HAN ; Ming XIANG ; Yiping LI ; Qing ZHANG ; Fei DAI ; Mingyue DENG
Chinese Journal of Orthopaedics 2024;44(14):956-962
Objective:To compare the clinical efficacy of rotator cuff repair combined with transposition of the long head of the biceps tendon in the treatment of massive rotator cuff tears with or without subscapularis injury.Methods:A total of 73 patients with massive rotator cuff tears who underwent arthroscopic rotator cuff repair combined with transposition of the long head of the biceps tendon in the Department of Upper Arm, Sichuan Orthopaedic Hospital from January 2019 to September 2022 were retrospectively analyzed, including 35 males and 38 females, aged 58.8±7.6 years (range, 46-73 years). There were 11 cases on the left side and 62 cases on the right side. Causes of injuries: 6 cases of car accidents, 13 cases of fall injuries, 54 cases of no obvious cause of injury. The patients were divided into subscapularis injury group (37 cases) and no subscapularis injury group (36 cases) according to the subscapularis injury or not. The visual analogue scale (VAS), Constant - Murley score, University of California Los Angeles (UCLA) shoulder score and range of motion were used to evaluate shoulder pain and functional improvement. MRI of the shoulder was taken at the final follow-up and the rotator cuff was assessed for retear using Sugaya grading.Results:All patients successfully completed the operation. The VAS score at 1 year postoperatively in the group without subscapularis injury was 0.58±0.87, which was less than 1.16±1.14 in the group with subscapularis injury ( t=-2.426, P=0.018). The Constant-Murley score and UCLA score at 1 year postoperatively in the group without subscapularis injury were 94.03±6.19 and 32.94±2.59, respectively, which were greater than those of the group with subscapularis injury, which were 86.51±11.96 and 30.38±3.33, and the difference was statistically significant ( P<0.05). Anterior flexion and body external rotation at 1 year postoperatively in the group without subscapularis injury were 137.22°±26.79° and 54.72°±13.20°, which were greater than those in the group with subscapularis injury of 121.08°±22.83° and 38.65°±17.19°, and the difference was statistically significant ( P<0.05), and the difference between the two groups in body internal rotation was not statistically significant ( P>0.05). The subacromial spacing at 1 year postoperatively was 9.4±1.3 mm in the group without subscapularis injury and 9.0±1.5 mm in the group with subscapularis injury, and the difference was not statistically significant ( t=1.318, P=0.192). There were 2 cases of rotator cuff retear 1 year after surgery without subscapular muscle injury and 5 cases of subscapular muscle injury. Conclusion:The clinical efficacy of arthroscopic rotator cuff repair combined with transposition of the tendon of the long head of the biceps tendon in the treatment of massive rotator cuff tears with or without subscapularis muscle injury is satisfactory, and the improvement of pain and function is more obvious in patients without subscapularis muscle injury.
5.Qingshen granules regulate mitophagy through miR-4516/SIAH3/PINK1 to attenuate renal fibrosis in rats
Kejun REN ; Hua JIN ; Dong WANG ; Yanping MAO ; Rong DAI ; Meng CHENG ; Yiping WANG ; Lei ZHANG
Chinese Journal of Pathophysiology 2024;40(8):1479-1487
AIM:To explore the mechanism by which Qingshen granules(QSG)intervene in the microRNA-4516(miR-4516)targeted regulation of the SIAH3/PINK1 axis,enhancing mitophagy and inhibiting renal fibrosis.METHODS:Male SD rats were randomly divided into three groups:control,model,and QSG groups.The QSG aqueous solution was administered via gavage once daily,4 mL each time,for 8 consecutive weeks.Blood creatinine levels were measured in each group.Hematoxylin-eosin(HE)and Masson staining were utilized to assess the degree of renal patholog-ical damage.Western blot analysis was performed to determine the expression levels of β-actin,PINK1,Parkin,SIAH3,VDAC1,Mfn1,Mfn2,OPA1,LC3B,and P62 proteins in renal tissue.RT-qPCR was used to detect the mRNA expres-sion level of SIAH3 in rat kidney tissue,and transmission electron microscopy was employed to observe mitochondrial dam-age in renal tissue.QSG-containing serum and transforming growth factor-β1(TGF-β1)were used to induce an HK-2 cell fibrosis model.The cells were divided into the following groups:normal cell(NC)group,model cell(MC)group,MC+miR-4516 mimics group,MC+miR-4516 NC+QSG group,MC+miR-4516 mimics+QSG group,and MC+QSG group.Cell activity in each group was detected using the CCK-8 method,and Western blot analysis was performed to determine E-cad-herin and α-SMA protein expression levels.The regulation of SIAH3 by miR-4516 was verified using a dual luciferase re-porter assay.RT-qPCR was used to detect the mRNA expression of miR-4516,SIAH3 mRNA,and PINK1.RESULTS:The results indicated that QSG intervention reduced fibrosis in rat renal tissue and HK-2 cells,decreased SIAH3 mRNA expression,increased PINK1 expression,and activated mitophagy in renal tissue.In vitro results confirmed that QSG can elevate miR-4516 expression,inhibit SIAH3 mRNA expression,promote PINK1 expression in HK-2 cells,and reduce the expression of the fibrosis marker protein α-SMA.CONCLUSION:In summary,this study preliminarily clarified the mechanism by which QSG intervention targets miR-4516 to regulate the SIAH3/PINK1 axis,thereby enhancing mitophagy and inhibiting renal fibrosis.
6.Common types and treatment of elbow injuries in skiing or snowboarding
Jinsong YANG ; Ming XIANG ; Yiping LI ; Qing ZHANG ; Fei DAI
Chinese Journal of Orthopaedic Trauma 2023;25(8):676-683
Objective:To investigate the common types of elbow joint injuries in skiing or snowboarding and their treatment.Methods:A retrospective study was conducted to analyze the data of 90 patients with elbow injury caused by skiing or snowboarding who had been admitted to Department of Upper Limbs, Sichuan Orthopaedic Hospital from February, 2015 to February, 2022. There were 53 males and 37 females with an age of (31.8±8.4) years. The types of elbow injury, visual analogue scale (VAS), range of motion (ROM) of the elbow and Mayo elbow performance score (MEPS) of the patients were recorded before treatment and at the last follow-up.Results:Of the patients, 18 were treated conservatively (3 simple Mason type I radial head fractures and 15 O'Driscoll type I coronal process fractures) and 72 surgically. Their follow-up time was (31.3±18.7) months. The fractures all healed by the time of the last follow-up. Simple elbow fracture was observed in 63 patients, simple elbow dislocation in 2 patients, fracture plus dislocation in 25 patients. The posterior dislocation was the most common (22 cases). Among the elbow fractures, radial head fracture occurred in 27 patients, ulnar coronoid process fracture in 41 patients, proximal ulnar fracture in 13 patients, and distal humeral fracture in 28 patients. Collateral ligament injuries were complicated in 65 cases. In the patients undergoing conservative treatment, their VAS, elbow ROM, and MEPS were all improved significantly from 4.7±1.4, 92.2°±14.4° and 63.9±6.5 before treatment to 0.4 (0,1.0), 110.6°±0.6°, and 92.2±3.9 at the last follow-up ( P<0.05); in the patients undergoing surgical treatment, their VAS, elbow ROM, and MEPS were also all improved significantly from 5.6±1.7, 24.3°±18.4°, and 26.9±12.2 before surgery to 0.6 (0,1.0), 97.4°±14.0° and 86.6±7.1 at the last follow-up ( P<0.05). After surgery, 8 patients presented with neurological symptoms and 7 patients developed heterotopic ossification. Conclusions:In skiing or snowboarding, the coronoid process of the ulna is the most vulnerable to fracture. In the elbow injuries due to skiing or snowboarding, posterior dislocation is the most common type which is often accompanied by injuries to the medial and lateral collateral ligaments. For simple Mason type Ⅰ radial head fractures and O'-Driscoll type Ⅰ coronoid process fractures in which the elbow is stable, conservative treatment can be adopted; surgical treatment is indicated for the other injuries.
7.Analysis of the supervision status of intensive countywide medical community in Fujian province based on thematic frame analysis
Zhao SHANG ; Dongyu XUE ; Yiping ZHENG ; Yueping LI ; Yue DAI
Chinese Journal of Hospital Administration 2023;39(6):411-416
Objective:To analyze the supervision mode and problems of intensive countywide medical communities and explore the methods and strategies to improve their supervision system.Methods:From March 2022 to May 2022, a stratified random sampling method was used and the intensive countywide medical communities in the province were selected for field study. In-depth interviews were made with the staff of the healthcare administration departments and health insurance agencies affiliated with intensive countywide medical communities, directors of general hospitals, directors of township hospitals, and heads of functional departments. These interviews mainly covered the principal entities of countywide medical community supervision, supervision measures and contents, supervision effects, supervision weaknesses, and how to improve internal and external supervision mechanisms. The interviews results were analyzed using thematic framework analysis and the data were subject to a descriptive analysis.Results:23 persons of five intensive countywide medical communities were selected for key person interview. The finalized framework of regulatory elements of intensive countywide medical communities consists of such five general themes as regulatory subjects, regulatory content, regulatory measures, regulatory effectiveness and existing problems, comprising 14 level-1 indicators and 52 level-2 indicators. The main regulatory bodies were mainly government departments (15 times), the regulatory content was focused on medical resources (10 times), the regulatory measures cover specific regulations (4 times), specific activities (4 times) and assessment and evaluation (4 times). And the regulatory effectiveness demonstrated a currently strong regulation strength of medical communities (5 times), and the existing problems were insufficient government supervision (12 times), difficulties encountered by social forces in their supervision (11 times) and internal governance (5 times).Conclusions:The supervision system of intensive countywide medical communities should be optimized by improving the governmental supervision mechanism, inviting supervision of social forces and building an effective internal governance.
8.Comparison of reverse shoulder arthroplasty and hemiarthroplasty in the treatment of three- or four-part fractures of proximal humeral in the elderly
Fei DAI ; Jingming DONG ; Jinsong YANG ; Qing ZHANG ; Yiping LI ; Ming XIANG
Chinese Journal of Orthopaedics 2022;42(4):204-212
Objective:To compare the clinical effects of reverse shoulder arthroplasty and hemiarthroplasty in the treatment of three- or four-part proximal humeral fractures in the elderly.Methods:The clinical data of 58 elderly patients with three- or four-part proximal humeral fractures treated with hemiarthroplasty or reverse shoulder arthroplasty from June 2014 to June 2020 were retrospectively analyzed. Among them, 46 cases were from Sichuan Provincial Orthopaedic Hospital (22 cases of hemiarthroplasty and 24 cases of reverse shoulder arthroplasty), and 12 cases were from Tianjin Hospital (5 cases of hemiarthroplasty and 7 cases of reverse shoulder arthroplasty). In the hemiarthroplasty group, there were 27 patients, including 7 males and 20 females, with an average age of 70.29±6.81 years (range, 61-87 years), and there were 10 cases of 3-part fractures and 17 cases of 4-part fractures. In the reverse shoulder arthroplasty group, including 9 males and 22 females, with an average age of 75.06 ±4.25 years (range, 67-86 years), and there were 9 cases of 3-part fractures and 22 cases of 4-part fractures. The postoperative efficacy evaluation indexes included visual analogue scale (VAS), range of motion (ROM), prosthesis upward displacement, healing of greater tuberosity, scapular glenoid notch, American Shoulder and Elbow Surgeons (ASES) and Constant-Murley score.Results:The average follow-up was 50.63±16.02 months (range, 24-75 months) in the hemiarthroplasty group and 28.32±11.93 months (range, 14-56 months) in the reverse shoulder arthroplasty group. The anterior elevation in the reverse shoulder arthroplasty group was 118.22°±27.22°, and those in the hemiarthroplasty group was 102.77°±25.88°, which was significant difference ( t=2.21, P=0.032); the results of external rotation (ER) and internal rotation (IR) in two groups were similar, and no significant difference (ER: t=0.57, P=0.616; IR: χ 2=2.61, P=0.273); the average Constant-Murley and ASES in the reverse shoulder arthroplasty group were significantly better than those in the hemiarthroplasty group ( P=0.019 and 0.018); the complication rates of hemiarthroplasty group and reverse shoulder arthroplasty group were 37% (10/27) and 13% (4/31), respectively (χ 2=4.59, P=0.032). In the hemiarthroplasty group, 6 patients had upward movements of the prosthesis and 2 patients had wear of the glenoid side; notching (sirveaux grade 1) was noted in 1 patient in the reverse shoulder arthroplasty group. Conclusion:In the treatment of three- or four-part proximal humeral fractures in the elderly, reverse shoulder arthroplasty achieves significantly better functional results compared to hemiarthroplasty.
9.Circular RNA expression profiles of peripheral blood mononuclear cells by microarray analysis in gout patients
Fei DAI ; Jianxiong ZHENG ; Yiping TANG ; Zengrong DONG ; Wenjun ZHOU ; Ting YI ; Quanbo ZHANG ; Yufeng QING
Chinese Journal of Rheumatology 2021;25(1):26-31,c1-3,c1-4
Objective:To analyze the expression of circular RNA (circRNA) in peripheral blood mononuclear cells (PBMCs) of patients with gout and to explore the possible mechanism of circRNA in the pathogenesis of gout.Methods:Peripheral blood samples of 24 patients with acute gout (AG), 24 patients with intermittent gout (IG) and 24 healthy control subjects (HC) were collected. Three cases of AG, IG, and HC were randomly selected, and the differentially expressed circRNA in PBMCs was screened by human circNA microarrays. The 6 circRNAs with large differences between the two comparison groups were selected, and the relative expression levels of 6 circRNAs in all the collected 72 PBMCs of the study subjects were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The significantly differentially ex-pressed circRNA (fold change>1.5, P<0.05) was analyzed by GO analysis, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and its interaction with microRNA (miRNA) was predicted. The median (interquartile range) was used to describe the data, and the Mann-Whitney U test was used for comparison between groups. Results:(1) The microarray analysis results showed that compared with the HC group, the AG group and the IG group had 116 and 41 significantly differently expressed circRNAs, respectively; com-pared with the IG group, the AG group had 105 significantly differently expressed circRNAs. (2) Among the 6 circRNAs verified by PT-qPCR, the expression trends of 5 were consistent with the microarray results. The expression of hsa_circRNA_105034 in the AG group [5.17(4.60)] was statistically significantly different com-pared to the IG [1.68(2.39)] and HC [0.90(0.73)] groups (AG vs IG: Z=-4.413, P<0.01; AG vs HC Z=-5.052, P<0.01). (3) Bioinformatics analysis: ① GO analysis found that differential circRNA swere mainly involved in DNA transcriptional regulation, positive cell regulation and protein modification, etc. ② KEGG pathway analysis revealed that differential circRNA might be involved in the immune response mediated by the mitogen-activated protein kinase signaling pathway. ③ CircRNA might affect its inflammatory response by targeting molecules such as miRNA-146a, miRNA-302b and miRNA-23a. Conclusion:There are differentially expressed circRNAs in PBMCs of patients with gout, which may be closely related to the occurrence and development of gout.
10.Study on the expression profile of circular RNA in patients with ankylosing spondylitis
Yiping TANG ; Ting YI ; Jianxiong ZHENG ; Fei DAI ; Zengrong DONG ; Quanbo ZHANG ; Yufeng QING
Chinese Journal of Rheumatology 2021;25(5):323-326,C5-2,C5-3
Objective:To screen for circle RNA (circRNA) differentially expressed in peripheral blood mononuclear cells (PBMCs) of patients with ankylosing spondylitis (AS), and to analyze its expression profile to explore the role of circRNAs in the pathogenesis of AS.Methods:CircRNA microarray chip technology was used to detect the expression of circRNAs in PBMCs of 3 patients with active AS, 3 patients with stable AS and 3 healthy controls (HC), and then screening for differentially expressed circRNAs by fold change (FC) and P value. Then differentially expressed circRNAs among the circRNAs with the highest differential expression were selected randomly to verify the chip results by real-time fluorescence quantitative polymerase chain reaction (qPCR); Differentially expressed circRNAs were subjected to Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and microRNA (miRNA) target prediction software was used to predict the circRNA/miRNA interaction relationship. Finally, the data were statistically analyzed by t test and Mann-Whitney U test. Results:① Chip text results showed that there were 800 circRNAs with significantly different expression (FC>1.5, P<0.05) in active AS than HC, of which 466 were up-regulated and 334 were down-regulated; the stable AS had a total of 1 149 significantly differentially expressed when compared with the HC (FC>1.5, P<0.05) circRNAs, of which 589 were up-regulated and 560 were down-regulated. 233 circRNAs were significantly differentially expressed (FC>1.5, P<0.05) between active AS and stable AS, of which 145 were up-regulated and 88 were down-regulated. ②The RT-qPCR verification results suggested that the expression trends of the four differentially expressed circRNAs were consistent with the results of the chip. ③ GO analysis results suggested that these differentially expressed circRNAs were mainly involved in nonsense-mediated mRNA decay, Rho GTPase binding and other processes. The analysis showed that the KEGG pathway were enriched in Th17 cell differentiation and chemokine signaling pathways. The results of miRNA target prediction software analysis suggested that differentially expressed circRNAs might play a role by targeting miR-650, let-7b-5p and other miRNAs. Conclusion:Compared with HC group, there were differentially expressed circRNAs in Peripheral Blood Mononuclear Cells (PBMCs) of AS patients, The results of this study suggest that these circRNAs may be involved in the pathogenesis of AS.


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