1.Establishment of a rapid diagnostic method to detect the presence of intracellular infection with Staphylococcus aureus
Luyao WEN ; Minghang LI ; Qijun ZHOU ; Jianhui WANG ; Shuai LIAN ; Rui WU ; Jianfa WANG
Chinese Journal of Veterinary Science 2025;45(9):1937-1943
Staphylococcus aureus has the ability to invade bovine mammary epithelial cells and cause persistent intracellular infections.For diagnosing mastitis in cows caused by the bacterial in-fections,it is necessary to determine whether there is a persistent intracellular infection by S.au-reus within mammary epithelial cells,and to establish scientific treatment strategies and imple-ment proactive management decisions.The content of this study is the establishment of a brand new diagnostic method aimed at confirming this type of bovine disease efficiently and cost-effec-tively.The diagnostic procedure is as follows:Add 100 μL S.aureus liquid culture with a concentra-tion of 8.5 ×1010 CFU/mL to the well-grown bovine mammary epithelial cell line and co-culture at 37 degrees Celsius for 2 h to establish an intracellular infection model;After the co-cultivation is completed,add 100 μL of lysostaphin to the dish to thoroughly eliminate extracellular S.aureus.It can be confirmed that the model has been successfully established that black spherical bacteria with a diameter of approximately 1μm are observed within the sample cells using a transmission e-lectron microscope.Intracellular infection is evidenced by the observation of DNA-staining positive material within the cytoplasm of DAPI-stained sample cells,as observed through a laser confocal microscope.The sample is subjected to gram staining after being treated with 200 μL of 0.5%Tri-tonX-100 for 15 min,and intracellular infection is observed within the cytoplasm of the sample cells with Gram-positive results under oil immersion microscopy.The results of the novel"Tri-tonX-100 & gram staining method"validate that the presence of Staphylococcus aureus individu-als within bovine mammary epithelial cells is consistent with images observed under transmission electron microscopy and laser confocal microscopy.The verification results indicate that the"Tri-tonX-100 & gram staining method"is suitable for the diagnosis of S.aureus persistent intracellu-lar infection in bovine mastitis.Later,somatic cells from milk were extracted by gradient centrifu-gation and purified to obtain bovine mammary epithelial cells for clinical testing of 70 S.aureus positive mastitis cattle,which showed a positive rate of 82.9%and a negative rate of 17.1%.
2.Exon Sequencing of HNF1β in Chinese Patients with Early-Onset Diabetes
Siqian GONG ; Hong LIAN ; Yating LI ; Xiaoling CAI ; Wei LIU ; Yingying LUO ; Meng LI ; Si-min ZHANG ; Rui ZHANG ; Lingli ZHOU ; Yu ZHU ; Qian REN ; Xiuying ZHANG ; Jing CHEN ; Jing WU ; Xianghai ZHOU ; Xirui WANG ; Xueyao HAN ; Linong JI
Diabetes & Metabolism Journal 2025;49(2):321-330
Background:
Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients.
Methods:
Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines.
Results:
Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher.
Conclusion
MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.
3.Exon Sequencing of HNF1β in Chinese Patients with Early-Onset Diabetes
Siqian GONG ; Hong LIAN ; Yating LI ; Xiaoling CAI ; Wei LIU ; Yingying LUO ; Meng LI ; Si-min ZHANG ; Rui ZHANG ; Lingli ZHOU ; Yu ZHU ; Qian REN ; Xiuying ZHANG ; Jing CHEN ; Jing WU ; Xianghai ZHOU ; Xirui WANG ; Xueyao HAN ; Linong JI
Diabetes & Metabolism Journal 2025;49(2):321-330
Background:
Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients.
Methods:
Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines.
Results:
Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher.
Conclusion
MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.
4.Exon Sequencing of HNF1β in Chinese Patients with Early-Onset Diabetes
Siqian GONG ; Hong LIAN ; Yating LI ; Xiaoling CAI ; Wei LIU ; Yingying LUO ; Meng LI ; Si-min ZHANG ; Rui ZHANG ; Lingli ZHOU ; Yu ZHU ; Qian REN ; Xiuying ZHANG ; Jing CHEN ; Jing WU ; Xianghai ZHOU ; Xirui WANG ; Xueyao HAN ; Linong JI
Diabetes & Metabolism Journal 2025;49(2):321-330
Background:
Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients.
Methods:
Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines.
Results:
Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher.
Conclusion
MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.
5.Exon Sequencing of HNF1β in Chinese Patients with Early-Onset Diabetes
Siqian GONG ; Hong LIAN ; Yating LI ; Xiaoling CAI ; Wei LIU ; Yingying LUO ; Meng LI ; Si-min ZHANG ; Rui ZHANG ; Lingli ZHOU ; Yu ZHU ; Qian REN ; Xiuying ZHANG ; Jing CHEN ; Jing WU ; Xianghai ZHOU ; Xirui WANG ; Xueyao HAN ; Linong JI
Diabetes & Metabolism Journal 2025;49(2):321-330
Background:
Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients.
Methods:
Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines.
Results:
Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher.
Conclusion
MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.
6.Establishment of a rapid diagnostic method to detect the presence of intracellular infection with Staphylococcus aureus
Luyao WEN ; Minghang LI ; Qijun ZHOU ; Jianhui WANG ; Shuai LIAN ; Rui WU ; Jianfa WANG
Chinese Journal of Veterinary Science 2025;45(9):1937-1943
Staphylococcus aureus has the ability to invade bovine mammary epithelial cells and cause persistent intracellular infections.For diagnosing mastitis in cows caused by the bacterial in-fections,it is necessary to determine whether there is a persistent intracellular infection by S.au-reus within mammary epithelial cells,and to establish scientific treatment strategies and imple-ment proactive management decisions.The content of this study is the establishment of a brand new diagnostic method aimed at confirming this type of bovine disease efficiently and cost-effec-tively.The diagnostic procedure is as follows:Add 100 μL S.aureus liquid culture with a concentra-tion of 8.5 ×1010 CFU/mL to the well-grown bovine mammary epithelial cell line and co-culture at 37 degrees Celsius for 2 h to establish an intracellular infection model;After the co-cultivation is completed,add 100 μL of lysostaphin to the dish to thoroughly eliminate extracellular S.aureus.It can be confirmed that the model has been successfully established that black spherical bacteria with a diameter of approximately 1μm are observed within the sample cells using a transmission e-lectron microscope.Intracellular infection is evidenced by the observation of DNA-staining positive material within the cytoplasm of DAPI-stained sample cells,as observed through a laser confocal microscope.The sample is subjected to gram staining after being treated with 200 μL of 0.5%Tri-tonX-100 for 15 min,and intracellular infection is observed within the cytoplasm of the sample cells with Gram-positive results under oil immersion microscopy.The results of the novel"Tri-tonX-100 & gram staining method"validate that the presence of Staphylococcus aureus individu-als within bovine mammary epithelial cells is consistent with images observed under transmission electron microscopy and laser confocal microscopy.The verification results indicate that the"Tri-tonX-100 & gram staining method"is suitable for the diagnosis of S.aureus persistent intracellu-lar infection in bovine mastitis.Later,somatic cells from milk were extracted by gradient centrifu-gation and purified to obtain bovine mammary epithelial cells for clinical testing of 70 S.aureus positive mastitis cattle,which showed a positive rate of 82.9%and a negative rate of 17.1%.
7.Feasibility study of selective stentless management after ureteroscopic lithotripsy
Li JIANG ; Yue WU ; Hua HU ; Lian FU ; Chaoying LIU ; Chao MA ; Gang ZHOU ; Yunhan RAN ; Rui ZHU
Chongqing Medicine 2025;54(11):2616-2620
Objective To investigate the feasibility of selectively omitting ureteral stent placement after ureteroscopic lithotripsy(URL).Methods A total of 118 patients with distal ureteral calculi undergoing URL from 2021 to 2024 were enrolled.Patients were divided into a control group(indwelling ureteral stent for 2 weeks,n=86)and an observation group(no ureteral stent placement,n=32).General data,operation time,hospital stay,and total medical costs were compared between the two groups.Patients were followed 2 weeks postoperatively for assessment of flank pain visual analogue scale(VAS)scores,bladder irritation symptoms,hematuria,and incidence of urinary tract infection.Hydronephrosis was evaluated by ultrasonography 3 months after surgery.Results There was no significant difference in the general information and operation time between the two groups(P>0.05).The length of hospital stay and total treatment cost in the observa-tion group were significantly lower than those in the control group(P<0.05).Two weeks after surgery,the VAS scores of low back pain on the affected side and occurrence rates of bladder irritation symptoms,hematu-ria,and urinary tract infection in the observation group were significantly lower than those in the control group(P<0.01).Three months after operation,no hydronephrosis was observed in both groups.Conclusion It is safe and feasible to avoid indwelling ureteral stent after URL in appropriate cases.
8.Efficacy of cosmetic suturing techniques combined with topical recombinant human basic fibroblast growth factor in repairing facial trauma
Huibin LIAN ; Pingsong LI ; Kunjie RONG ; Gang XU ; Jie ZHOU ; Yi LUO ; Zhongyin ZHUFU ; Haibin JU ; Junjun JIN ; Rui JIAO ; Xiaowei ZHANG ; Zhimin YIN
Chinese Journal of Medical Aesthetics and Cosmetology 2024;30(6):571-575
Objective:To observe the efficacy of cosmetic suturing techniques combined with topical recombinant human basic fibroblast growth factor (rh-bFGF) in repairing facial trauma.Methods:A prospective study was conducted on 140 patients with facial trauma admitted to the Department of Burn and Plastic Surgery, Northern Jiangsu People's Hospital from January to December 2022. Patients were divided into two groups based on different treatment methods using a random number table method: treatment group (70 cases), including 38 males and 32 females aged 3 to 54 (23.1±8.2) years, received cosmetic suturing techniques combined with topical rh-bFGF for wound repair; control group (70 cases), including 36 males and 34 females aged 2 to 49 (22.3±7.5) years, only received cosmetic suturing techniques for wound repair. Patients were followed up 2 weeks post-surgery to evaluate wound healing quality. Patient satisfaction was assessed using the visual analogue scale (VAS). Six months post-surgery, scar conditions were evaluated using the Vancouver scar scale (VSS).Results:In the treatment group, 65 cases were directly sutured, and 5 cases were repaired with skin flaps, with a first-class healing rate of 100% (70/70). In the control group, 66 cases were directly sutured, and 4 cases were repaired with skin flaps, with a first-class healing rate of 91.4% (64/70). The first-class healing rate in the treatment group was higher than that in the control group, with a statistically significant difference ( P=0.037). Two weeks post-surgery, the VAS score for surgical satisfaction in the treatment group was (1.13±0.52) scores, which was lower than that in the control group (2.56±1.32) scores, with a statistically significant difference ( P<0.001). Six months post-surgery, the VSS score for the treatment group was (2.49±1.27) scores, which was lower than that in the control group (4.67±1.93) scores, with a statistically significant difference ( P<0.001). Conclusions:In repairing facial trauma, the combination of cosmetic suturing techniques and topical rh-bFGF can improve wound healing quality, reduce wound scarring, and enhance patient satisfaction with surgery.
9.MRI-based radiomics machine learning model for differentiating small renal cell carcinoma from fat-poor renal angiomyolipoma
Rui-Ting WANG ; Lian-Ting ZHONG ; Xian-Pan PAN ; Lei CHEN ; Meng-Su ZENG ; Yu-Qin DING ; Jian-Jun ZHOU
Chinese Journal of Clinical Medicine 2023;30(6):940-945
Objective To investigate the value of multi-phase MRI-based radiomics machine learning models in differentiating small renal cell carcinoma(sRCC)from fat-poor renal angiomyolipoma(fp-AML).Methods 79 cases of sRCCs and 35 cases of fp-AMLs(diameter≤4 cm)which were confirmed by pathology were retrospectively analyzed.The volume of interest(VOI)of the total tumor was manually delineated on the images of T2WI(T2),unenhanced phase(UP),corticomedullary phase(CMP)and nephrographic phase(NP)and then the radiomics of the VOIs were extracted respectively.The training set and the test set were set according to the ratio of 7∶3.The t-test,maximal relevance and minimal redundancy(mRMR)and the least absolute shrinkage and selection operator(LASSO)were used to select the radiomics features.The selected features were used to build classification models with logistic regression(LR)and support vector machine(SVM).The receiver operating characteristic(ROC)curve was used to evaluate the classification performances of the models.Results There were 4,12,3,11 and 15 optimal features obtained from T2、UP、CMP、NP and the combined four phases,respectively.The radiomics features based on NP or the combined four phases with LR model performed best,AUCs were respectively 0.956,0.986 in the training set and both were 0.881 in the test set.Conclusion The multi-phase MRI-based radiomics machine learning model has favorable diagnostic performance in differentiating sRCC from fp-AML.
10.Effects of Compound Danshen Dripping Pills on Ventricular Remodeling and Cardiac Function after Acute Anterior Wall ST-Segment Elevation Myocardial Infarction (CODE-AAMI): Protocol for a Randomized Placebo-Controlled Trial.
Yu-Jie WU ; Bo DENG ; Si-Bo WANG ; Rui QIAO ; Xi-Wen ZHANG ; Yuan LU ; Li WANG ; Shun-Zhong GU ; Yu-Qing ZHANG ; Kai-Qiao LI ; Zong-Liang YU ; Li-Xing WU ; Sheng-Biao ZHAO ; Shuang-Lin ZHOU ; Yang YANG ; Lian-Sheng WANG
Chinese journal of integrative medicine 2023;29(12):1059-1065
BACKGROUND:
Ventricular remodeling after acute anterior wall ST-segment elevation myocardial infarction (AAMI) is an important factor in occurrence of heart failure which additionally results in poor prognosis. Therefore, the treatment of ventricular remodeling needs to be further optimized. Compound Danshen Dripping Pills (CDDP), a traditional Chinese medicine, exerts a protective effect on microcirculatory disturbance caused by ischemia-reperfusion injury and attenuates ventricular remodeling after myocardial infarction.
OBJECTIVE:
This study is designed to evaluate the efficacy and safety of CDDP in improving ventricular remodeling and cardiac function after AAMI on a larger scale.
METHODS:
This study is a multi-center, randomized, double-blind, placebo-controlled, parallel-group clinical trial. The total of 268 patients with AAMI after primary percutaneous coronary intervention (pPCI) will be randomly assigned 1:1 to the CDDP group (n=134) and control group (n=134) with a follow-up of 48 weeks. Both groups will be treated with standard therapy of ST-segment elevation myocardial infarction (STEMI), with the CDDP group administrating 20 tablets of CDDP before pPCI and 10 tablets 3 times daily after pPCI, and the control group treated with a placebo simultaneously. The primary endpoint is 48-week echocardiographic outcomes including left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume index (LVEDVI), and left ventricular end-systolic volume index (LVESVI). The secondary endpoint includes the change in N terminal pro-B-type natriuretic peptide (NT-proBNP) level, arrhythmias, and cardiovascular events (death, cardiac arrest, or cardiopulmonary resuscitation, rehospitalization due to heart failure or angina pectoris, deterioration of cardiac function, and stroke). Investigators and patients are both blinded to the allocated treatment.
DISCUSSION
This prospective study will investigate the efficacy and safety of CDDP in improving ventricular remodeling and cardiac function in patients undergoing pPCI for a first AAMI. Patients in the CDDP group will be compared with those in the control group. If certified to be effective, CDDP treatment in AAMI will probably be advised on a larger scale. (Trial registration No. NCT05000411).
Humans
;
ST Elevation Myocardial Infarction/therapy*
;
Stroke Volume
;
Ventricular Remodeling
;
Prospective Studies
;
Microcirculation
;
Ventricular Function, Left
;
Myocardial Infarction/etiology*
;
Treatment Outcome
;
Percutaneous Coronary Intervention/adverse effects*
;
Heart Failure/drug therapy*
;
Drugs, Chinese Herbal/therapeutic use*
;
Randomized Controlled Trials as Topic
;
Multicenter Studies as Topic

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