1.Behavior of cartilage-derived microtissue and ability of cartilage formation in three-dimensional dynamic and static culture conditions
Wei LIU ; Hongyu JIANG ; Jiajie CHEN ; Yuyang GAO ; Yanjun GUAN ; Zhibo JIA ; Ying JIAO ; Zhen HUA ; Gehan JIANG ; Ying HE ; Aiyuan WANG ; Jiang PENG ; Jianhong QI
Chinese Journal of Tissue Engineering Research 2024;28(25):4022-4026
BACKGROUND:Compared with traditional two-dimensional culture,three-dimensional microtissue culture can show greater advantages.However,more favorable cultivation methods in three-dimensional culture still need to be further explored. OBJECTIVE:To evaluate the cell behavior of microtissue and its ability to promote cartilage formation under two three-dimensional culture methods. METHODS:Cartilage-derived microcarriers were prepared by chemical decellularization and tissue crushing.DNA quantification and nuclear staining were used to verify the success of decellularization,and histological staining was used to observe the matrix retention before and after decellularization.The microcarriers were characterized by scanning electron microscopy and CCK-8 assay.Cartilage-derived microtissues were constructed by combining cartilage-derived microcarriers with human adipose mesenchymal stem cells through three-dimensional static culture and three-dimensional dynamic culture methods.The cell viability and chondrogenic ability of the two groups of microtissues were detected by scanning electron microscopy,live and dead staining,and RT-qPCR. RESULTS AND CONCLUSION:(1)Cartilage-derived microcarriers were successfully prepared.Compared with before decellularization,the DNA content significantly decreased after decellularization(P<0.001).Scanning electron microscope observation showed that the surface of the microcarrier was surrounded by collagen,maintaining the characteristics of the natural extracellular matrix of cartilage cells.CCK-8 assay indicated that microcarriers had no cytotoxicity and could promote cell proliferation.(2)Scanning electron microscopy and live and dead staining results showed that compared with the three-dimensional static group,the three-dimensional dynamic group had a more extended morphology of microtissue cells,and extensive connections between cells and cells,between cells and matrix,and between matrix.(3)The results of RT-qPCR showed that the expressions of SOX9,proteoglycan,and type Ⅱ collagen in microtissues of both groups were increased at 7 or 14 days.The relative expression levels of each gene in the three-dimensional dynamic group were significantly higher than those in the three-dimensional static group at 14 days(P<0.05).At 21 days,the three-dimensional static group had significantly higher gene expression compared with the three-diomensional dynamic group(P<0.001).(4)The results showed that compared with three-dimensional static culture microtissue,three-dimensional dynamic culture microtissue could achieve higher expression of chondrogen-related genes in a shorter time,showing better cell viability and chondrogenic ability.
2.LncRNA LUCAT1 Promotes the Pathogenesis of Intrauterine Adhesion by Regulating AREG
Jianhong WU ; Yucui TIAN ; Zi-Wen JIANG
Journal of Medical Research 2024;53(5):92-98
Objective To investigate the effect of long non-coding RNA(lncRNA)lung cancer-related transcript 1(LUCAT1)on the pathogenesis of intrauterine adhesion by regulating the expression of amphiregulin(AREG),to provide a new molecular target for the prevention and treatment of intrauterine adhesions.Methods Real-time quantitative polymerase chain reaction(RTqPCR)was used to determine the mRNA expression levels of lncRNA LUCAT1,fibrotic markers of α-smooth muscle actin(α-SMA)and collagen type Ⅰ alpha 1 chain protein(COL1A1)in samples of intrauterine adhesion tissue and endometrial stromal cell treated with transforming growth factor-β1.Western blot was used to determine the protein expression levels of AREG,α-SMA,COL1A1.Then,si-LUCAT1,pcDNA LUCAT1,si-AREG were transfected into ESC and RT-qPCR was used to detect the mRNA expression levels of lncRNA LUC-AT1 and AREG.Next,si-LUCAT1 and pcDNA LUCAT1 were transfected into ESC and treated these cells with TGF-β1 for 48h,re-spectively.Western blot was used to further detect the protein expression levels of AREG,α-SMA and COL1A1,and cell proliferation and apoptosis were detected by cell proliferation assay and cell apoptosis assay.Results The expression levels of lncRNA LUC AT1,AR-EG and fibrosis markers α-SMA and COL1A1 were upregulated in endometrial tissues from patients with intrauterine adhesion and in ESC that had been treated with TGF-β1.AREG changed with the change of lncRNA LUCAT1,and the expression of lncRNA LUC-AT1did not change significantly after downregulation of AREG,and AREG was positively regulated by lncRNA LUCAT1.During the process of the transformation of ESC into fibroblasts,si-LUCAT1 significantly inhibited the protein expression levels of AREG,α-SMA and COL1A1(P<0.01),significantly reduced cell proliferation and significantly induced cell apoptosis,the difference was statistically significant(P<0.001).pcDNA LUCAT1 significantly induced the protein expression levels of AREG,α-SMA and COL1A1,and the difference was statistically significant(P<0.01).Conclusion LncRNA LUCAT1 promotes the pathogenesis of intrauterine adhesion by up-regulating the expression of AREG.LncRNA LUCAT1/AREG axis may provide novel molecular target for the prevention and treat-ment of intrauterine adhesion.
3.Value of amide proton transfer-weighted imaging with intravoxel incoherent motion imaging for diagnosing and evaluating the differentiation of cervical squamous cell carcinoma
Zhonghong XIN ; Jianhong PENG ; Xiande LU ; Jiang NAN ; Yaping ZHANG ; Zixian CHEN ; Xiaohui WANG ; Jun ZHU ; Junqiang LEI
Chinese Journal of Radiology 2024;58(6):627-632
Objective:To explore the value of amide proton transfer-weighted (APTw) imaging and intravoxel incoherent motion (IVIM) imaging for diagnosing and evaluating the pathological differentiation of cervix squamous cell carcinoma (CSCC).Methods:This study was a diagnostic trial. Totally 56 patients pathologically diagnosed with CSCC at the First Hospital of Lanzhou University from October 2021 to October 2022 were retrospectively collected, as the CSCC group. And 36 female healthy volunteers who underwent physical examinations at the First Hospital of Lanzhou University from October 2021 to October 2023 were recruited as the control group. CSCC patients were divided into well-moderately differentiated ( n=34) and poorly differentiated groups ( n=22). The region of interest was placed in the lesions of CSCC group and normal cervical stroma of control group, and the quantitative parameters for asymmetric magnetization transfer ratio (MTR asym) of APTw imaging and pure diffusion coefficient (D), false diffusion coefficient (D *) and perfusion fraction (f) for IVIM were obtained. The independent sample t test was used to compare the differences in quantitative parameters between the two groups, the logistic regression model was used to establish combined parameters for the quantitative parameters with statistical significance between the two groups. The receiver operator characteristic curve was used to evaluate the diagnostic efficacy of single quantitative parameters and combined parameters to distinguish the CSCC group from the control group, and the well-moderately differentiated group from the poorly differentiated group in CSCC patients. The area under the curve (AUC) was compared using the DeLong test. Results:There were significant differences in MTR asym, D and f between CSCC group and control group ( t=-9.79, 10.09, 11.35, P<0.001). Also, significant differences were found for MTR asym and D between the well-moderately differentiated and poorly differentiated group ( t=4.11, -3.76, P<0.001). There was no significant difference in other quantitative parameters ( P>0.05). When comparing the CSCC group and control group, the AUC (95% CI) of MTR asym, D, f and combined parameter (MTR asym+D+f) were 0.887 (0.804-0.944), 0.940 (0.871-0.979), 0.968 (0.909-0.993), 0.995 (0.950-1.000). The AUC of the combined parameter was higher than those of MTR asym and D, with statistical significance ( Z=3.07, 2.06, P=0.002, 0.040). When comparing the well-moderately differentiated and poorly differentiated group, the AUC (95% CI) of MTR asym, D, and combined parameter (MTR asym+D) were 0.789 (0.660-0.887), 0.775 (0.644-0.876), 0.852 (0.731-0.932). There was no significant difference between each two AUCs ( P>0.05). Conclusion:The quantitative parameters of APTw and IVIM imaging can be used to diagnose and preliminarily evaluate the pathological differentiation of CSCC. Joint parameters can improve the diagnostic efficiency of CSCC.
4.Systematic review of factors related to quality of life in patients with oral cancer:a systematic review
Mei LIU ; Xinglian SHI ; Zhezhen LI ; Jianhong JIANG
West China Journal of Stomatology 2024;42(4):486-493
Objective This study aims to evaluate factors related to quality of life of patients with oral cancer.Methods CNKI,Wanfang,VIP,CBM,Pubmed,Medline,Web of Science,Embase,and The Cochrane Library were searched up to May 2023 for studies that evaluated the quality of life of patients with oral cancer.All the included studies were independently selected,extracted,and rated by two researchers,and results are summarized by qualitative analysis.Results Twenty-four articles on 2 717 patients were included.Factors related to the quality of life of patients with oral cancer mainly included age,tumor TNM stage,radiochemotherapy,and gender,which could be summarized into three as-pects:personal factors,disease-related factors,and surgical factors.More than five studies reported on the analysis of age,gender,tumor TNM stage,pathological stage,neck dissection method,marital status,recurrence,smoking,educa-tion level,etc.The results are relatively consistent.Conclusion The incidence of oral cancer increases,and many fac-tors affected the quality of life.The included literature is a cross-sectional study,and the sample size is limited.The caus-al relationship between relevant factors and quality of life should be verified using large sample sizes.
5.Practice of paperless management of medical records in a hospital
Ling HONG ; Weifeng YING ; Qin SUN ; Jianhong JIANG ; Liangfeng ZHU
Chinese Journal of Hospital Administration 2024;40(5):400-404
Paperless management of medical records is of great significance in promoting refined hospital management. Driven by national policies and the development of information technology in medical institutions, paperless medical records have become an inevitable trend in the development of medical records. However, at present, there were still a large number of medical record data in China that cannot be digitized or directly uploaded to hospital electronic medical record systems. Since 2019, the First Hospital of Jiaxing has launched a paperless medical record construction project and established a paperless medical record management system, forming a closed-loop management system for the entire process of medical record collection, archiving, storage, management, and utilization. By the end of 2021, paperless of medical records in all departments had been achieved, electronic signatures had been fully implemented, and paperless versions of reports of standalone equipment, high-value consumables barcodes, surgical anesthesia related documents, and outsourced inspection reports had been achieved, with a paperless rate of 98.5%. Through paperless management of medical records, the supervision of medical record quality and medical quality has been strengthened, operating costs have been reduced and work efficiency has been improved. At the same time, this has provided convenience for medical personnel to access medical record information and applicate of the data, and has effectively improved the hospital′s level of refined management. It can provide reference for paperless management of medical records in other medical institutions.
6.Clinical application of jejunal feeding tube for early enteral nutrition after surgical treatment of upper digestive tract malformation in newborns
Yuli ZHONG ; Lin ZHOU ; Xue JIANG ; Jianhong YAN ; Xing ZHOU ; Ling LIU
Chinese Journal of Neonatology 2023;38(3):146-150
Objective:To study the clinical application of jejunal feeding tube (J-tube) for early enteral nutrition after surgical treatment of upper digestive tract malformation in newborns.Methods:From January 2019 to December 2021, newborns with upper digestive tract malformation received stage Ⅰ small bowel resection and anastomosis in our hospital were enrolled in this prospective randomized controlled study. According to different types of postoperative nutritional support, these patients were randomly assigned into J-tube group and control group using block randomization method. The J-tube group were given enteral nutrition vis J-tube within 48-72 h after surgery and the control group were given oral feeding after the recovery of gastrointestinal function. Calories and proteins intake, growth indicators, duration of hospital stay and parenteral nutrition, time needed for full oral feeding and complications were compared between the two groups.Results:A total of 24 patients were in J-tube group and 28 in controlled group. No significant differences existed on the general status between the two groups ( P>0.05). The average daily intake of calories and proteins in j-tube group in the first week after surgery were significantly higher than control group [(108.7±8.3) kcal/(kg·d) vs. (97.9±7.0) kcal/(kg·d), (3.4±0.3) g/(kg·d) vs. (3.1±0.2) g/(kg·d)] ( P<0.05). No significant differences existed in the average daily intake of calories and proteins during the second postoperative week between the two groups ( P>0.05). Compared with control group,J-tube group showed increased growth velocity in head circumference and weight over time ( P<0.05), while the trend over time in length growth was not significant ( P>0.05). No significant differences existed in the duration of hospital stay and parenteral nutrition, time needed for full oral feeding and complications between the two groups ( P>0.05). Conclusions:Enteral nutrition via J-tube 48-72 h after surgery is safe and feasible in the postoperative nutritional management of newborns with upper digestive tract malformation. This strategy may promote physical growth after surgery without increasing the incidences of complications.
7.Expression of POU2F2 in clear cell renal cell carcinoma and its effects on the biological behavior of the cancer cells
Meiyu JIANG ; Xinhong ZHAO ; Yunfeng NIU ; Fanglong LI ; Jianhong QIU
Journal of Modern Urology 2023;28(6):529-535
【Objective】 To investigate the expression of transcription factor POU domain class 2 transcription factor 2 (POU2F2) in clear cell renal cell carcinoma (ccRCC) and human renal cancer cell lines (786-O and ACHN) and its effects on the cells’ biological behaviors such as proliferation, migration and invasion in vitro. 【Methods】 The mRNA expressions of POU2F2 in ccRCC tissues, adjacent normal tissues, cell lines 786-O and ACHN were detected with real-time polymerase chain reaction (qRT-PCR). The protein expression of POU2F2 in ccRCC tissues and adjacent normal tissues were detected with immunohistochemistry. The effects of knockdown of POU2F2 on the mRNA and protein expressions of epithelial mesenchymal transformation (EMT)-related tumor markers were detected with qRT-PCR and Western blot. 【Results】 The mRNA expression of POU2F2 in ccRCC tissues was significantly higher than that in adjacent normal tissues, and was correlated with patients’ gender, WHO/ISUP nuclear grade and TNM stage. The protein expression of POU2F2 was significantly higher in ccRCC tissues than in adjacent normal tissues, and was correlated with tumor pathological grade and TNM stage. The mRNA expression of POU2F2 was significantly decreased in 786-O cells after sh-POU2F2-1013 plasmid transfection (P<0.05); the proliferation ability, clonal formation rate, migration ability and invasion ability were significantly reduced (P<0.05). Knockdown of POU2F2 down-regulated the mRNA and protein expressions of MMP2, MMP9 and Twist in 786-O cells, while up-regulated E-ca expression. 【Conclusion】 The mRNA expression of POU2F2 was significantly up-regulated in ccRCC tissues and renal cancer cells. Knockdown of POU2F2 inhibited the proliferation, migration and invasion of cells in vitro, and slowed or inhibited the occurrence and development of renal cancer.
8.AGO and RDRP genes are involved in the stress response of Aspergillus flavus
Xiang Liu ; Bi Yang ; Xun Tian ; Jianhong Zhou ; Yonghui Liao ; Lingling Liu ; Wenfeng Yu ; Xiaolan Qi ; Yinhui Jiang
Acta Universitatis Medicinalis Anhui 2023;58(9):1442-1449
Objective :
To explore the role of Argonaute ( Ago) gene and RNA⁃Dependent RNA Polymerase (RDRP) gene of Aspergillus flavus in the growth and development about the RNAi mechanism .
Methods :
A. flavus Ago1 , Ago2 , RDRP1 , RDRP3 gene mutant strains were constructed by homologous recombination . The growth and development of the mutant strains were observed on potato dextrose agar(PDA) + uracil uridine (UU) medium inoculated with 3 μl 106 CFU/mL spores . 200 , 400 μg cell wall pressure agent conidored ( CR) , 0. 8 mol/L , 1 . 6 mol/L osmotic pressure agent NaCl , 2 mmol/L , 4 mmol/L oxidative pressure agent hydrogen peroxide (H2 O2 ) and 0. 01% , 0. 02% genomic damage agent methyl mesylate (MMS) were added to the Yeast extract Glucose Minimum (YGM) + UU medium to analyze the stress response of the mutant strains .
Results :
A. flavus mutant strains about ΔAgo1 , ΔAgo2 , ΔRDRP1 , ΔRDRP3 were successfully constructed and its growth and development were normal . The ΔAgo1 and ΔAgo2 strains reduced the stress effects on cell wall and osmotic pressure compared to the control . Ago1 gene deletion reduced the effect of H2 O2 , and conversely RDRP3 gene deletion increased the inhibition of H2 O2 . The Ago2 and RDRP1 strains reduced the effect on genetic damage agent . In addition , ΔRDRP1 increased the effect of osmotic stress .
Conclusion
The Ago1 , Ago2 , RDRP1 and RDRP3 genes of A. flavus are not in⁃ volved in the regulation of growth rate and asexual reproduction and can participate in the regulating of the host stress response to the environment .
9.Study on Inflammatory Immune Mechanism of Lactoferrin in the Treatment of Periodontitis
XU Junfeng ; XU Wanjun ; DONG Yanrong ; DENG Zuyue ; JIANG Xia ; YUAN Ying ; FANG Jianhong ; WAN Yue ; REN Yanyun
Chinese Journal of Modern Applied Pharmacy 2023;40(15):2086-2092
OBJECTIVE To study the anti-inflammatory immune response effects of lactoferrin in the treatment of periodontitis and its mechanism. METHODS One hundred SD rats were randomly divided into blank control group, model group, lactoferrin administration group low, medium, high dose group(1, 2, 3 g·kg-1), metronidazole positive control group (0.02 g·kg-1), PDTC group(200 mg·kg-1), lactoferrin+PDTC group(2 g·kg-1, 200 mg·kg-1), MCC950 group(1 mg·kg-1) and lactoferrin+MCC950 group(2 g·kg-1, 1 mg·kg-1), 10 rats in each group. Silk thread ligation combined with 10% sucrose drinking water was used to establish the model, and then the drug was administered orally once a day. The blank control group and the model group were administered orally with 0.9% NaCl. The rats in each group were sacrificed after one month of continuous administration. The contents of IL-1b, IL-8 and IL-10 were detected by ELISA kit, and the expressions of TLR2-NF-κB pathway and NLRP3 inflammasome related proteins were detected by Western blotting. HE staining was used to observe the pathological changes of the periodontal tissues of the rats in each group. RESULTS Compared with the model group, the symptoms of periodontitis in each dose group of lactoferrin were significantly improved. HE staining showed that the infiltration of inflammatory cells was reduced, and the proliferation of fibroblasts was active. The protein expressions of TLR2, NF-κB, NLRP3, Caspase-1 p20 and GSDMD-N decreased, the content of pro-inflammatory factor IL-8 and IL-1b decreased, and the content of anti-inflammatory factor IL-10 increased. CONCLUSION Lactoferrin may play a role in the regulation of inflammatory immune response in the treatment of periodontitis by down-regulating the protein expression of TLR2-NF-κB-NLRP3 pathway, reducing the initiation of inflammatory response and the release of inflammatory factors, so as to achieve the purpose of anti-inflammatory.


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