1.Effect of lncRNA-TNFRSF13C on hypoxia-inducible factor 1alpha in periodontal cells by modulation of miR-1246
Jing BAI ; Xue ZHANG ; Yan REN ; Yuehui LI ; Xiaoyu TIAN
Chinese Journal of Tissue Engineering Research 2025;29(5):928-935
BACKGROUND:LncRNA-TNFRSF13C,an important factor in B cell development and function,is expressed in periodontal tissues of patients with periodontitis,but the specific mechanism is still unclear. OBJECTIVE:To investigate the mechanism of lncRNA-TNFRSF13C regulating miR-1246 on hypoxia-inducible factor 1α in periodontal cells. METHODS:Human periodontal ligament cells(hPDLCs)were treated with lipopolysaccharide and divided into group A(hPDLCs cell lines without transfection),group B(hPDLCs cell lines transfected with TNFRSF13C NC-siRNA),group C(hPDLCs cell lines transfected with TNFRSF13C-siRNA),group D(hPDLCs cell line transfected with miR-1246 mimics),group E(hPDLCs cell line transfected with miR-1246 siRNA),group F(hPDLCs cell line transfected with TNFRSF13C-siRNA+miR-1246 mimics),and group G(hPDLCs cell line transfected with TNFRSF13C-siRNA+miR-1246 siRNA).The relative expression of lncRNA-TNFRSF13C and miR-1246 in each group was detected by qRT-PCR.Cell counting kit-8 assay was used to detect cell viability.Apoptosis was detected by flow cytometry.Expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins was detected by western blot.The correlation between lncRNA-TNFRSF13C and miR-1246 was analyzed by Pearson,and the targeting relationship was analyzed by dual-luciferase reporter assay. RESULTS AND CONCLUSION:There was no significant difference in human periodontal ligament cell activity,apoptosis rate and protein indexes between groups A and B(P>0.05).Compared with group B,hPDLCS cell activity in group C was increased,and apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins were decreased(P<0.05).Compared with group C,hPDLCS cell activity in group D was decreased,and apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins were increased(P<0.05).Compared with group D,the cell activity of group E was increased(P<0.05).The cell activity in group F was lower than that in group E,and the apoptosis rate was reduced in both groups E and F(P<0.05).Compared with group F,the cell activity of group G was increased,and the apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor were decreased(P<0.05).LncRNA-TNFRSF13C was positively correlated with miR-1246(P<0.05).Compared with the TNFRSF13C-siRNA group,the fluorescence activity of miR-1246-wt in the TNFRSF13C-NC group was reduced(P>0.05);compared with the miR-1246-NC group,the fluorescence activities of hypoxia-inducible factor 1α-wt and vascular endothelial growth factor-wt in the miR-1246 mimics group were increased(P<0.05).To conclude,down-regulation of lncRNA-TNFRSF13C can promote the activity of periodontal cells treated with lipopolysaccharide,reduce apoptosis,and inhibit hypoxia-inducible factor 1α and vascular endothelial growth factor.The mechanism is related to the regulation of miR-1246 activity.
2.Evaluation of FTA-LAMP direct extraction method for extracting DNA from Streptococcus mutans
Yuehui WANG ; Jin SHANG ; Chen YANG ; Dongge FU ; Can CAO ; Xiaodong ZHANG ; Jingfu WANG
Chinese Journal of Tissue Engineering Research 2025;29(5):1043-1049
BACKGROUND:Streptococcus mutans is an important pathogen of dental caries,and timely detection of its levels is of great significance for early detection and treatment of dental caries. OBJECTIVE:To evaluate the effect of loop-mediated isothermal amplification(FTA-LAMP)direct extraction of Streptococcus mutans DNA. METHODS:(1)Bacterial suspensions containing ATCC standard strains(Streptococcus mutans)were prepared and inoculated into the brain-heart leachate medium.After mixed thoroughly,the mixture was then diluted in a 10-fold gradient into seven concentrations(4.2×107,4.2×106,4.2×105,4.2×104,4.2×103,4.2×102,4.2×10 CFU/mL),two parallel controls were made for each dilution level,and sterile water was used as a blank control.(2)The DNA of Streptococcus mutans was extracted using FTA Elute card,boiling method,kit extraction and lysate extraction methods separately and then amplified using LAMP technology was amplified.A specificity test was also performed to compare the differences between the four DNA extraction methods.RESULTS AND CONCLUSION:The DNA extracted by all four methods met the requirements for LAMP amplification.Specificity test results showed that only Streptococcus mutans could specifically amplify the target gene.The detection limit value of the DNA concentration was 4.2×103 CFU/mL for the lysate method,4.2×104 CFU/mL for the FTA Elute card extraction method,4.2×106 CFU/mL for the kit extraction method,and 4.2×107 CFU/mL for the boiling method.In the other aspects of the four extraction methods,the kit extraction method had the highest experimental cost,number of steps and time;the other three methods had the same number of steps,with the FTA Elute card method requiring the least amount of instruments,the boiling method having the lowest single cost,and the lysate extraction method taking the least amount of time.Only a small amount of bacteria were needed for successful extraction using both the FTA Elute card and lysate extraction methods.Compared with the FTA Elute card method,the lysate extraction method was superior in terms of time,but it had a high single cost and required more equipment.To conclude,the FTA-LAMP technology established in this study has the advantages of ease of operation,high specificity,high sensitivity,and visualization,which is expected to be a new way for efficient extraction and detection of Streptococcus mutans.
3.Effect of Modified Shoutai Pill (寿胎丸加味方) on Inflammatory Reaction and Expression of Endometrial Receptivity-Related Factors in A Rat Model of Polycystic Ovary Syndrome and Miscarriage with High Testosterone-Insulin Resistance
Tingting GUO ; Meng JIANG ; Huaiying YANG ; Xiang JI ; Yuehui ZHANG
Journal of Traditional Chinese Medicine 2025;66(3):275-282
ObjectiveTo explore the possible mechanisms of Modified Shoutai Pill (寿胎丸加味方, MSP) in treating polycystic ovary syndrome (PCOS) with hyperandrogenism, insulin resistance, and miscarriage, focusing on inflammatory response and endometrial receptivity. MethodsThirty female SPF-grade SD rats with regular estrous cycles and in proestrus, and 15 male SPF-grade SD rats were housed together in a 2∶1 ratio at 18:00. At 8:00 next morning, rats showing abundant sperm and vaginal plugs were considered pregnant on the day 0.5. The 30 pregnant rats were randomly divided into three groups, normal group, model group, and MSP group, with 10 rats in each group. From day 0.5 to day 13.5 of pregnancy, the MSP group was given 26.6 g/(kg·d) of the MSP via gavage twice a day for 14 consecutive days. The normal group and the model group received 4 ml of normal saline daily. From day 7.5 to day 13.5 of pregnancy, the rats in the model group and MSP group were intraperitoneally injected with dihydrotestosterone (DHT) and insulin (INS) for 7 consecutive days to establish a PCOS model with hyperandrogenism, insulin resistance, and miscarriage. On day 13.5 of pregnancy, an oral glucose tolerance test (OGTT) was performed to measure blood glucose levels at 0, 30, 60, 90, and 120 minutes. On day 14.5, serum level of progesterone (P4), estradiol (E2), fasting insulin (FINS), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were measured by ELISA. The insulin resistance index (HOMA-IR) was calculated. Embryo implantation, miscarriage rate, and average number of live fetuses were observed. Uterine tissue pathology was examined by HE staining, and mRNA expression of Il-6, Tnf-α, leukemia inhibitory factor (Lif), homeobox gene 10 (Hoxa10), prolactin family 8 subfamily A member 2 (Prl8a2), and insulin-like growth factor-binding protein 1 (Igfbp1) in the uterine tissue was detected by qRT-PCR. ResultsCompared with the normal group, the model group had significantly higher blood glucose level at 0, 30, 60, 90, and 120 minutes, increased miscarriage rate, elevated HOMA-IR, decreased average number of live fetuses, lower level of P4 and E2, higher level of IL-6, TNF-α, and FINS, and higher mRNA expression of Il-6 and Tnf-α in the uterine tissue. The mRNA expression of Lif, Hoxa10, and Prl8a2 was reduced (P<0.05 or P<0.01). The uterus had a dark red color, visible areas of bleeding, fewer embryos with developmental abnormalities, and increased placental necrosis. Pathological examination revealed thrombus in the decidual layer, unclear decidual cell morphology, loose arrangement, scattered distribution, edema degeneration in the cytoplasm, and nuclear shrinkage or disappearance, with extensive infiltration of inflammatory cells. In contrast, compared with the model group, the MSP group showed significantly lower blood glucose level at 0, 30, 60, 90, and 120 min, reduced miscarriage rate, lower HOMA-IR, increased number of live fetuses, higher level of P4 and E2, and lower level of IL-6, TNF-α, and FINS. The mRNA expression of Il-6 and Tnf-α in the uterine tissue was lower, while the expression of Lif, Hoxa10, and Prl8a2 mRNA was higher (P<0.05 or P<0.01). There was significant improvement in uterine and embryo conditions, as well as in uterine tissue pathology. ConclusionThe MSP can reduce the miscarriage rate in a PCOS model with hyperandrogenism, insulin resistance, and miscarriage. Its mechanism may involve inhibiting inflammation, improving endometrial receptivity, and restoring the defects in endometrial decidualization.
4.Itaconic acid improves vascular endothelial dysfunction in diabetic mice by inhibiting endothelial cell apoptosis
Jia TANG ; Yuehui YIN ; Yunlin CHEN ; Jian YANG ; Fuwei ZHANG ; Luning WANG
Journal of Chongqing Medical University 2025;50(9):1195-1201
Objective:To investigate the protective effect and mechanism of 4-octyl itaconate(4-OI)on vascular endothelial function in diabetic mice,since endothelial dysfunction is the key and initial factor for vascular complications in diabetes.Methods:The db/db mice were used to establish a mouse model of diabetes,and high glucose(HG)-induced human umbilical vein endothelial cells(HUVECs)were used to establish a cell model of diabetes.After 4-OI treatment,HE staining,Masson staining,and reticular fiber staining were used to observe pathological changes of the thoracic aorta in the control group,the model group,and the intervention group;Western blot was used to measure the expression of α-smooth muscle actin(α-SMA),a marker for endothelial fibrosis;immuno-fluorescence assay was used to measure the expression levels of Bcl-2-associated X protein(BAX)and B-cell lymphoma-2(Bcl-2)in the thoracic aorta of mice in the control group,the model group,and the intervention group;immunoblotting was used to observe the effect of 4-OI intervention on the expression of BAX,Bcl-2,and the proteins associated with the cytochrome C(CytC)/cysteinyl aspartate specific proteinase(caspase)pathway in HG-induced HUVECs.Results:The results showed that 4-OI could significantly improve vascular endothelial dysfunction in db/db diabetic mice.Compared with the model group,the intervention group had a significant increase in the expression of Bcl-2(0.73±0.07 vs.0.52±0.04,F=49.380,P=0.009)and a significant reduction in the expression of BAX(1.41±0.14 vs.1.89±0.12,F=37.270,P=0.008)after 4-OI treatment.In the cell model of HUVECs induced by HG,compared with the high glucose group,the intervention group had a sig-nificant increase in the expression of Bcl-2(1.22±0.04 vs.0.81±0.09,F=75.410,P<0.001)and a significant reduction in the expres-sion of BAX(0.83±0.04 vs.1.12±0.04,F=268.100,P<0.001)after 4-OI treatment,as well as significant reductions in the expres-sion of CytC(0.61±0.03 vs.1.01±0.03,F=234.000,P<0.001),caspase-9(0.62±0.03 vs.1.04±0.04,F=164.300,P<0.001),and caspase-3(0.92±0.06 vs.1.74±0.04,F=424.100,P<0.001).Conclusion:In conclusion,4-OI inhibits vascular endothelial cell apoptosis in diabetic mice by affecting the expression of the CytC/Caspase pathway factors,thereby improving vascular endothelial dys-function in diabetes.
5.Application and Implications of Surgical Complications Indicators from the Perspective of Hospital Performance Evaluation
Yuehui LIU ; Shunfei LI ; Li MAO ; Tianyi ZHANG
Chinese Hospital Management 2025;45(12):31-34
Surgical complications,as a key focus and challenge in hospital quality management,serve as important indicators for medical quality monitoring and hospital performance evaluation.From the perspective of hospital performance evaluation,it reviews relevant indicators related to surgical complications in domestic and international hospital performance evaluation systems.lt analyzes the definition and assessment methods of surgical complications,the calculation and evaluation methods of surgical complication incidence rates,and the association between surgical complications and medical insurance payments.The aim is to provide reference guidelines for further improving China's monitoring and evaluation system for surgical complications,thereby enhancing the accuracy,scientific rigor,and fairness of performance evaluations
6.WANG Xiuxia's Clinical Experience in Treating Hyperprolactinemia with Liver Soothing Therapy
Yu WANG ; Danni DING ; Yuehui ZHANG ; Songli HAO ; Meiyu YAO ; Ying GUO ; Yang FU ; Ying SHEN ; Jia LI ; Fangyuan LIU ; Fengjuan HAN
Journal of Traditional Chinese Medicine 2025;66(14):1428-1432
This paper summarizes Professor WANG Xiuxia's clinical experience in treating hyperprolactinemia using the liver soothing therapy. Professor WANG identifies liver qi stagnation and rebellious chong qi (冲气) as the core pathomechanisms of hyperprolactinemia. Furthermore, liver qi stagnation may transform into fire or lead to pathological changes such as spleen deficiency with phlegm obstruction or kidney deficiency with essence depletion. The treatment strategy centers on soothing the liver, with a modified version of Qinggan Jieyu Decoction (清肝解郁汤) as the base formula. Depending on different syndrome patterns such as liver stagnation transforming into fire, liver stagnation with spleen deficiency, or liver stagnation with kidney deficiency, heat clearing, spleen strengthening, or kidney tonifying herbs are added accordingly. In addition, three paired herb combinations are commonly used for symptom specific treatment, Danggui (Angelica sinensis) with Chuanxiong (Ligusticum chuanxiong), Zelan (Lycopus lucidus) with Yimucao (Leonurus japonicus) , and Jiegeng (Platycodon grandiflorus) with Zisu (Perilla frutescens).
7.Construction of a rapid and quantitative platform for loop-mediated isothermal amplification and smart-phone-based detection of Cariogenic microorganisms
Jin SHANG ; Xiaodong ZHANG ; Jingfu WANG ; Yuehui WANG ; Chen YANG ; Heliang WANG ; Jiao-jiao LIU ; Ran ZHANG
Journal of Practical Stomatology 2025;41(5):595-599
Objective:To establish a rapid,quantitative and visualized method for the rapid detection of Streptococcus mutans(S.mutans)based on loop-mediated isothermal amplification(LAMP).Methods:Nucleic acid of S.mutans was extracted for LAMP reaction,the detection experimental conditions were optimized.Finally the positive results of the experiments were quantita-tively analyzed by using the mobile-based detection equipment.Results:The system detected the samples within 23 min and showed good specificity and sensitivity with an accuracy of 1.625×10-3 ng/μL.Positive results were captured,identified and quantified u-sing a mobile application,and the quantified results showed a good linear relationship between the absolute G value and the DNA concentration,with an equation of y=3.2x+57.133(R2=0.988 2).Conclusion:Quantitative detection of S.mutans by LAMP has the characteristics of high sensitivity,high specificity,result visualization,convenience and time-saving,and can quantify bacterial DNA.It is a new method for the early detection of caries susceptibility,and can be widely used in clinic.
8.Construction of a rapid and quantitative platform for loop-mediated isothermal amplification and smart-phone-based detection of Cariogenic microorganisms
Jin SHANG ; Xiaodong ZHANG ; Jingfu WANG ; Yuehui WANG ; Chen YANG ; Heliang WANG ; Jiao-jiao LIU ; Ran ZHANG
Journal of Practical Stomatology 2025;41(5):595-599
Objective:To establish a rapid,quantitative and visualized method for the rapid detection of Streptococcus mutans(S.mutans)based on loop-mediated isothermal amplification(LAMP).Methods:Nucleic acid of S.mutans was extracted for LAMP reaction,the detection experimental conditions were optimized.Finally the positive results of the experiments were quantita-tively analyzed by using the mobile-based detection equipment.Results:The system detected the samples within 23 min and showed good specificity and sensitivity with an accuracy of 1.625×10-3 ng/μL.Positive results were captured,identified and quantified u-sing a mobile application,and the quantified results showed a good linear relationship between the absolute G value and the DNA concentration,with an equation of y=3.2x+57.133(R2=0.988 2).Conclusion:Quantitative detection of S.mutans by LAMP has the characteristics of high sensitivity,high specificity,result visualization,convenience and time-saving,and can quantify bacterial DNA.It is a new method for the early detection of caries susceptibility,and can be widely used in clinic.
9.Application and Implications of Surgical Complications Indicators from the Perspective of Hospital Performance Evaluation
Yuehui LIU ; Shunfei LI ; Li MAO ; Tianyi ZHANG
Chinese Hospital Management 2025;45(12):31-34
Surgical complications,as a key focus and challenge in hospital quality management,serve as important indicators for medical quality monitoring and hospital performance evaluation.From the perspective of hospital performance evaluation,it reviews relevant indicators related to surgical complications in domestic and international hospital performance evaluation systems.lt analyzes the definition and assessment methods of surgical complications,the calculation and evaluation methods of surgical complication incidence rates,and the association between surgical complications and medical insurance payments.The aim is to provide reference guidelines for further improving China's monitoring and evaluation system for surgical complications,thereby enhancing the accuracy,scientific rigor,and fairness of performance evaluations
10.Discussion on the medication law of TCM compound patents in treating polycystic ovary syndrome
Yaoman WANG ; Lei GAO ; Yuehui ZHANG
International Journal of Traditional Chinese Medicine 2024;46(3):377-382
Objective:To analyze the medication and compatibility law of TCM compounds with national patents for the treatment of polycystic ovary syndrome (PCOS); To provide reference for the new TCM prescriptions for the treatment of PCOS.Methods:TCM compound patents for PCOS were retrieved from the China National Intellectual Property Administration's Chinese Patent Announcement Website from the establishment of the website to April 20, 2023. SPSS 25.0, IBM SPSS Modeler 18.0, and Cytoscape 3.8.0 were used to carry out frequency statistics, association rule analysis on the data, and a TCM core complex network and systematic clustering analysis were built.Results:126 compound patents were included, involving 392 kinds of Chinese materia medica, with a total frequency of 1 709 times. The medicinal property was mainly warm, the taste was mainly sweet, and the meridian was mainly liver meridian. High frequency drugs included Angelica Sinensis (46 times), Cuscutae Semen (44 times), Cyperi Rhizoma (42 times), Rehmanniae Radix Praeparata (41 times), Salviae Miltiorrhizae Radix et Rhizoma (37 times), etc. Commonly used medicinal pairs included Poria-Atractylodis Macrocephalae Rhizoma (support 16.67%, confidence 76.19%), Angelica Sinensis-Paeoniae Radix Alba (support 15.87%, confidence 80.00%), etc. The triple drug combinations were Cyperi Rhizoma-Pinelliae Rhizoma-Poria (support 12.70%, confidence 81.25%). The core prescriptions included "Poria, Pinelliae Rhizoma, Citri Rettculatae Pericarpium, Cyperi Rhizoma, Salviae Miltiorrhizae Radix et Rhizoma, Epimedii Folium, Atractylodis Macrocephalae Rhizoma, Morindae Officinalis Radix, Gleditsiae Spina, Acori Tatarinowii Rhizoma", etc. Potential prescriptions were "Hordei Fructus Germinatus, Setariae Fructus Germinatus, Sargentodoxae Caulis, Puerariae Lobatae Radix, Leonuri Fructus" and so on.Conclusion:The treatment of PCOS with TCM compounds with national patents mainly focuses on drying dampness and strengthening the spleen, tonifying kidney yang and nourishing kidney yin, promoting blood circulation.

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