1.Effect of chromosome-instability-associated gene GALNT7 on proliferation and apoptosis of HCT116 colon cancer cells
Xiao Fang ; Wei Zhao ; Wenying Yu ; Lingjie Pei ; Wenxuan Qian ; Ya Zhao
Acta Universitatis Medicinalis Anhui 2025;60(1):96-101
		                        		
		                        			Objective :
		                        			To investigate the effect of chromosome instability(CIN) associated gene polypeptide N-acetylgalactosaminyltransferase 7(GALNT7) on proliferation and apoptosis of HCT116 colon cancer cells.
		                        		
		                        			Methods :
		                        			The HCT116 cell line withGALNT7knockdown was constructed by lentiviral infection. The correlation betweenGALNT7and CIN was verified by chromosome spread assay. The effect ofGALNT7on cell proliferation was detected by live cell counting, and the effect ofGALNT7on cell cycle distribution was detected by flow cytometry and Western blot. Caspase-3 activity and Western blot assays were used to detect the effect ofGALNT7on apoptosis. 
		                        		
		                        			Results :
		                        			HCT116 cells showed a slower proliferation rate upon knocking down ofGALNT7, and exhibited a more scattered karyotype distribution and a phenotype of increased degree of CIN. Inhibition ofGALNT7in HCT116 cells resulted in cell cycle arrest, upregulation of P21 and downregulation of CDK6 protein levels, as well as increased levels of Caspase-3 activity, cleaved PARP1 and PUMA protein expression, and decreased levels of BCL-2 protein expression. 
		                        		
		                        			Conclusion 
		                        			TheGALNT7gene may promote proliferation and inhibit apoptosis of HCT116 colon cancer cells through the suppression of CIN generation.
		                        		
		                        		
		                        		
		                        	
2.Study of the mechanism of acupuncture and moxibustion in protecting the intestinal mucosal barrier in DSS-induced UC rats based on the IL-9/IL-9R pathway
Yan HUANG ; Kexin SUN ; Jing XU ; Zhe MA ; Jimeng ZHAO ; Lingjie LI ; Chen ZHAO ; Ling YANG ; Feng LI ; Yanan LIU ; Yue ZHANG ; Luyi WU ; Rude HUANG
Journal of Acupuncture and Tuina Science 2024;22(2):91-103
		                        		
		                        			
		                        			Objective:To observe the effects of acupuncture and moxibustion on interleukin(IL)-9/IL-9 receptor(IL-9R)in the colon tissue of rats with ulcerative colitis(UC)and investigate the protective mechanism of acupuncture and moxibustion on the intestinal mucosal barrier in UC rats. Methods:Male Sprague-Dawley rats were randomly divided into a normal control(NC)group and a modeling group.UC models were prepared by giving 4%dextran sulfate sodium(DSS)water for 7 d.After the successful construction of the UC rat model,the modeling group was randomly divided into a UC group,a herb-insulated moxibustion(HM)group,and an electroacupuncture(EA)group.HM and EA interventions at bilateral Tianshu(ST25)were performed once a day for 7 d.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes in the colon.The serum concentrations of IL-9,IL-6,IL-1β,and hemoglobin-H(HbH)were determined by enzyme-linked immunosorbent assay.The protein expression levels of IL-9,IL-9R,claudin-2,zonula occludens-1(ZO-1),and occludin in the colon tissue were measured by Western blotting or immuno-histochemistry.Immunofluorescence was used to detect the co-expression of PU.1 and CD4 with the IL-9 protein. Results:Compared with the NC group,the colon tissue of UC rats was severely damaged and ulcerated with congestion and edema,and the colonic histopathological score increased significantly(P<0.01).The serum HbH concentration decreased significantly(P<0.01),while the serum concentrations of IL-9,IL-6,and IL-1β increased(P<0.01).The protein expression of colonic ZO-1 and occludin decreased significantly(P<0.01),while the protein expression of colonic IL-9 and IL-9R increased(P<0.05).The positive co-expression levels of IL-9/PU.1 and IL-9/CD4 increased in the colon tissue(P<0.05).Compared with the UC group,the colonic mucosal structures were gradually repaired in both HM group and EA group,and healed ulcers could be observed,the colonic histopathological score decreased significantly(P<0.05).The serum concentration of HbH increased(P<0.01),while the serum concentrations of IL-9,IL-6,and IL-1β decreased(P<0.05).The protein expression levels of ZO-1 and occludin increased(P<0.05),while the protein expression levels of IL-9 and IL-9R decreased(P<0.01).The positive co-expression levels of IL-9/PU.1 and IL-9/CD4 decreased in the colon tissue(P<0.05). Conclusion:Both HM and EA can inhibit the protein expression levels of IL-9 and IL-9R in the UC colon by regulating the transcription factor PU.1,promote the repair of intestinal mucosal barrier,and down-regulate protein contents of proinflammatory factors IL-9,IL-6,and IL-1β in the serum,which may be one of the key mechanisms of acupuncture and moxibustion in reducing the inflammation of UC colonic mucosa and protecting the intestinal mucosal barrier.
		                        		
		                        		
		                        		
		                        	
3.Analysis of the efficacy of root canal cleaning by PTG and TFA root canal treatment systems
Haiou SUN ; Jiaojiao LIU ; Lingjie ZHAO
Journal of China Medical University 2024;53(8):747-751
		                        		
		                        			
		                        			Objective To compare the cleaning abilities of ProTaper Gold(PTG)and TF-Adaptive(TFA).Methods Forty single-abstracted premolars were randomly divided into PTG and TFA groups.The debris and smear layer of the root coronal 1/3,middle 1/3 and apical 1/3 in the two groups were observed using scanning electron microscope(SEM).The cleaning abilities of the PTG and TFA rotary systems were scored according to the scoring standard developed by YAMASHITA.The scores of the two groups were compared and then analyzed.Results There was no significant difference found in the scores of the smear layer and debris at the root coronal 1/3 and the middle 1/3.The scores of the debris and smear layer at the apical 1/3 in the group PTG were higher than those in the TFA group(P<0.05).Conclusion PTG and TFA can effectively remove the root canal smear layer and debris at the root coronal 1/3,and the middle 1/3.However,at the apical 1/3,the cleaning efficiency of TFA is better than that of PTG.
		                        		
		                        		
		                        		
		                        	
4.Application of patient-based real-time quality control using exponentially weighted moving average method on quality control procedures of thyroid function tests
Xiaoqing ZHAO ; Aijun NIU ; Xiaoling WANG ; Lingjie REN ; Haiting MAO
Chinese Journal of Laboratory Medicine 2024;47(5):526-535
		                        		
		                        			
		                        			Objective:To investigate the application of patient-based real-time quality control (PBRTQC) using exponentially weighted moving average (EWMA) method in internal quality control (IQC) procedures of thyroid function tests.Methods:The serum thyroid function test results of outpatients and inpatients in the Second Hospital of Shandong University from December 1, 2022 to April 30, 2023 were collected. Based on the PBRTQC professional intelligent software system, normality correction, parameter setting, program preparation and real-time operation of test data were carried out. The results of all patients who underwent thyroid function testing between May 1, 2023 and August 31, 2023 were used as the validation dataset. The estimated EWMA value of thyroid function test results and the cumulative coefficient of variation ( CV) over 4 months were calculated. The cumulative CV was compared with the criteria of precision quality standard (1/3TEa) and the CV of IQC. Westgard 2-2s and 1-3s rules were used for alarm setting. The early warning information of the EWMA quality control program were recorded and the potential causes of performance changes were analyzed. DxLab Mind software was used to conduct normal distribution statistics for all data, and the Kolmogorov-Smirnov test was performed on the test results. Results:The items related to serum thyroid function of the patients were all positively skewed. After data correction by Box-Cox method, the PBRTQC data of free triiodothyronine (FT3) and free thyroxine (FT4) were normally distributed, and their cumulative precisions ( CV) of EWMA within 4 months were 6.26% and 2.86%, respectively, both of which were lower than the precision quality target of 8.33%. However, the data of thyroid-stimulating hormone (TSH), thyroid peroxidase antibodies (TPOAb) and thyroglobulin antibody (TgAb) were still positive skewed after modification. The EWMA cumulative CV of TSH, TPOAb and TgAb were 13.16%, 15.31% and 16.77%, which were higher than the precision quality targets of 8.33%, 10% and 10%, respectively. The EWMA QC program can detect different out-of-control alarms, including FT4 false alarms due to sample source concentration and TSH result bias caused by changes in reagent performance. In addition, the EWMA QC program can also detect differences in FT3 results between different DXI800 fully automated chemiluminescence instrument instruments. Conclusions:The EWMA program based on PBRTQC professional intelligent software tools can monitor the patient data of the detection system in real time and continuously, dynamically identify and monitor the errors generated during the analysis process and give early warning. It can be used as a useful supplement for the daily IQC of thyroid function items, especially FT3 and FT4, and has good clinical application value.
		                        		
		                        		
		                        		
		                        	
5.Tangeretin inhibits tumor stemness of non-small cell lung cancer by regulating PI3K/AKT/mTOR signaling pathway
Sai WANG ; Lingjie WANG ; Yanli LI ; Peng LI ; Mengjun LI ; Donghua ZHAO ; Yongjie WANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2023;30(04):614-621
		                        		
		                        			
		                        			Objective    To study the effect of Tangeretin on non-small cell lung cancer (NSCLC) and the tumor stemness, and to find the molecular mechanism of its effect. Methods    We used cell counting and cell cloning experiments to study the effect of Tangeretin on the proliferation of NSCLC cells in vitro. The effect of Tangeretin on the invasion of NSCLC cells was detected by transwell assay. We detected the effect of Tangeretin on the proliferation of NSCLC cells in vivo by nude mouse tumor-bearing experiment. The effect of Tangeretin on tumor stemness of NSCLC cells was detected by self-renew assay, and CD133 and Nanog protein expressions. The expressions of PI3K/AKT/mTOR signaling pathway-related proteins were detected by Western blotting (WB). Results    Tangeretin had a good inhibitory effect on the proliferation of NSCLC cells in vivo and in vitro. Cell counting experiment, clonal formation experiment and nude mouse tumor-bearing experiment showed that Tangeretin could inhibit the proliferation activity, clonal formation ability, and tumor size of NSCLC cells in vivo. Self-renew experiments showed that Tangeretin could inhibit the self-renew ability of NSCLC cells. WB experiments showed that Tangeretin inhibited the expressions of tumor stemness markers CD133 and Nanog in NSCLC cells. Tangeretin could inhibit the activation of PI3K/AKT/mTOR signaling pathway-related proteins in NSCLC cells, and the activation of PI3K/AKT/mTOR signaling pathway could partially remit the inhibitory  effect of Tangeretin on tumor stemness of NSCLC cells. Conclusion    Tangeretin can inhibit the tumor stemness of NSCLC cells, which may be related to the regulation of PI3K/AKT/mTOR signaling pathway.
		                        		
		                        		
		                        		
		                        	
6.Telmisartan affects proliferation, migration and apoptosis of non-small cell lung cancer cell A549 through the Wnt/β-catenin signaling pathway
Lingjie WANG ; Donghua ZHAO ; Zhangfeng HUANG ; Mengjun LI ; Pengfei GUO ; Yongjie WANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(01):100-105
		                        		
		                        			
		                        			Objective    To investigate the effects of telmisartan on the proliferation, migration and apoptosis of non-small cell lung cancer A549 and the mechanism of regulating Wnt signaling pathway. Methods    Non-small cell lung cancer cell line A549 was cultured in vitro. Cell counting kit-8 (CCK-8) assay was used to detect the effect of telmisartan at different concentrations on the proliferative activity of A549 cells. The survival fraction of A549 treated with different concentrations of telmisartan was determined by colony-formation assay. The effect of telmisartan at different concentrations on the migration ability of A549 cells was examined in the wounding healing assay. Hoechst staining was used to detect the effects of telmisartan at different concentrations on the apoptosis of A549. Western bloting was used to detect the expressions of β-actin, proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, Wnt-3a, Beta-catenin (β-catenin), serine protein kinase 3β (p-GSK-3β), glycogen synthase kinase-3β (GSK-3β) and c-myc. Results    Different concentrations of telmisartan treatment inhibited the proliferation activity, colony-formation rate and migration of A549 cells, and reduced the expression of PCNA in a concentration-dependent manner. Telmisartan treatment promoted the apoptosis of A549 cells, significantly increased the expression of pro-apoptotic protein Bax and decreased the expression of anti-apoptotic protein Bcl-2. The expression levels of Wnt-3a, β-catenin, p-GSK-3β, and c-myc in A549 cells increased after treatment with telmisartan, while the expression levels of GSK-3β decreased. Conclusion    Telmisartan may play a role in the proliferation, migration and apoptosis of non-small cell lung cancer A549 cells, and  inhibiting the Wnt/β-catenin signaling pathway may be one of the mechanisms.
		                        		
		                        		
		                        		
		                        	
7.Lunch satisfaction and leftovers among students of Changsha compulsory education schools
XU Lingjie, QIU Fuping, CHEN Zhiqiu, ZHAO Haiyang, GUO Shiyin, MA Guansheng
Chinese Journal of School Health 2021;42(2):198-202
		                        		
		                        			Objective:
		                        			To understand lunch satisfaction and leftovers of Changsha compulsory education schools under different supply modes served by school canteens and specialized enterprises,and to provide scientific basis for the government to formulate school lunch improvement policies.
		                        		
		                        			Methods:
		                        			Multistage stratified cluster sampling method was used to random select 2 203 students from 8 primary and secondary schools in Changsha who were administered with questionnair survey in April to May of 2018.
		                        		
		                        			Results:
		                        			For school lunch, the overall rate of satisfaction was 78.8%.The overall lunch satisfaction of female and primany school students were higher than that of male and junior high school students respectively,and the difference was statistically significant(P<0.05). Totally 76.0% of students had the highest satisfaction in food hygiene and the lowest satisfaction rate was 60.0% in food taste. By comparing different supply modes, the overall satisfaction rate of meal quantity was higher (47.8%) under school canteens, and that of dining environment was relatively high (43.7%) under specialized enterprises. "Don-t like some food" (63.8%) was the main reason for students leftovers under the two supply modes. The lunch leftover rate of specialized enterprises (37.8%) was significantly higher than that of school cafeteria (30.6%)(χ 2=12.81,P<0.05).
		                        		
		                        			Conclusion
		                        			The satisfaction of food taste and the rate of lunch leftovers need to be further improved. School canteens environment and management should be strengthened, as well as the flexibility and communication under specialized enterprises and family-school communication. The joint efforts by school and family are needed for healthy eating education.
		                        		
		                        		
		                        		
		                        	
8.Clinical significances of additional chromosome abnormalities and t (15;17) in acute promyelocytic leukemia
Yingying LIU ; Tingting FU ; Chunting ZHAO ; Boli CUI ; Lingjie SUN ; Zhan SU ; Xiaodan LIU ; Tianlan LI
Journal of Leukemia & Lymphoma 2019;28(2):100-103
		                        		
		                        			
		                        			Objective To investigate the clinical significances of additional chromosome abnormalities and t(15;17) in acute promyelocytic leukemia (APL). Methods A total of 90 newly diagnosed APL patients in the Affiliated Hospital of Qingdao University from January 2007 to June 2014 were analyzed retrospectively. Patients with different chromosome karyotypes were divided into four groups: additional chromosome number abnormalities group (16 cases), additional chromosome structural abnormalities group (14 cases), additional chromosome number and structural abnormalities group (4 cases) and typical chromosome group (56 cases). According to whether the patient contained t(15;17), the patients were divided into group with t (15;17) and group without t (15;17). The short-term efficacy and survival of each group were analyzed and compared. Results The rate of complete remission in additional chromosome number abnormalities group, additional chromosome structural abnormalities group, additional chromosome number and structural abnormalities group and typical t(15;17) chromosome changes group were 56.3%(9/16), 100.0%(14/14), 25.0%(1/4) and 82.1%(46/56), the early mortality rates were 25.0%(4/16), 0 (0/14), 75.0%(3/4) and 8.9% (5/56) respectively. Among them, the additional number and structural abnormalities group had lower complete remission rate and higher early mortality rate, and compared with other groups, the differences were statistically significant (all P< 0.05). The complete remission rates of the group with t (15;17) and the group without t (15;17) were 80.5% (66/82) and 50.0% (4/8), respectively, and the difference was not statistically significant (P= 0.070). Conclusions APL patients with karyotypes with additional number and structural changes have low complete remission rate, high early mortality rate and poor prognosis. Patients with t(15;17)have a high rate of complete remission.
		                        		
		                        		
		                        		
		                        	
9.A study of the frequencies of HLA-DQA1 alleles and their clinical application values in the donor-recipient HLA-10/10 matched hematopoietic stem cell transplantation
Qinqin ZHAO ; Yang LI ; Zhijuan PAN ; Lingjie LI ; Luyao CHEN ; Jun HE
Chinese Journal of Microbiology and Immunology 2015;35(12):926-931
		                        		
		                        			
		                        			Objective To analyze the frequencies of HLA-DQA1 alleles and their clinical values in the donor-recipient HLA-A,-B,-C,-DRB1,-DQB1 (10/10) matched hematopoietic stem cell transplantation (HSCT).Methods This study recruited 127 patients who received allogeneic HSCT and 127 unrelated donors.High-resolution (High Res) DNA typing for HLA-DQA1 alleles were performed on the 254 subjects by using sequence specific oligonucleotide probes (SSOP) and high resolution of sequence specific primer(High Res SSP).Results The DQA1 allele genotypes of 36 pairs of donor-recipient were directly identified by using SSOP.The ambiguous DQA1 allele genotypes of the rest 91 pairs were identified by using High Res SSP.Among the 127 pairs of donor-recipient,5 pairs were HLA-DQA1 alleles mismatched,while the others were all matched.No significant differences in the distribution of HLA-DQA1 alleles were observed between the donors and the recipients.Sixteen HLA-DQA1 alleles were detected in the 127 donors,which were DQA1 * 02 ∶ 01 (19.3%),DQA1* 01 ∶ 02(19.3%),DQA1 * 03 ∶ 02/03 (17.0%),DQA1 *01∶03 (9.8%),DQA1*06∶01(9.1%),DQA1*05∶ 01(7.1%),DQA1*05∶05(5.9%),DQA1*03∶01 (4.7%),DQA1*01 ∶04(2.4%),DQA1*01∶05(2.0%),DQA1*01∶01(1.2%),DQA1*05 ∶ 03(0.8%),DQA1 *05 ∶ 08(0.8%),DQA1*04 ∶ 01(0.4%),DQA1*05 ∶ 06(0.4%) from high to low frequency.Moreover,a new allele was detected in the patients.The haplotypes' frequencies and linkage disequilibrium(LD) analysis of HLA-DQA1 and HLA-DQB1 showed that the most common haplotype was DQA1 *02 ∶ 01-DQB1 *02 ∶ 02(16.1%),followed by DQA1 *03 ∶ 02/03-DQB1 *03 ∶ 03 (11.8%)and DQA1 *01 ∶ 03-DQB1 * 06 ∶ 01 (9.1%).Stronger LD were observed between DQA1 * 02 ∶ 01 and DQB1*02 ∶ 02,DQA1 *03 ∶ 02 and DQB1*03 ∶ 03,DQA1 *01 ∶ 03 and DQB1*06 ∶ 01,HLA-DQA1*06∶01 andDQB1*03 ∶ 01,DQA1*05 ∶ 01 and DQB1*02 ∶ 02(P<0.001).Conclusion There was strong linkage disequilibrium between HLA-DQA1 and HLA-DQB1 genes.The polymorphism of HLA-DQA1 gene was less than that of HLA-DQB1 gene.No more guidance was provided to donor selection in unrelated donor-recipient HLA matched HSCT by adding HLA-DQA1 genotyping,but it might have clinical application values in HSCT with HLA Ⅱ locus mismatched donor and recipient.
		                        		
		                        		
		                        		
		                        	
10.Effect of curcumin on expression of synovial osteoprotegerin and receptor activator of nuclear factor-κB ligand protein in adjuvant arthritis rats
Yan QIU ; Wei SHANG ; Zhiming ZHAO ; Junhao GUO ; Hui CAI ; Lingjie ZHAO
Chinese Journal of Immunology 2014;(11):1490-1493,1498
		                        		
		                        			
		                        			Objective:By observing the effects of curcumin on synovial pathology, synovial osteoprotegerin and receptor activator of nuclear factor κB ligand expression factor of adjuvant arthritis rat models, we try to explore the possible mechanism of prevention and treatment of rheumatoid arthritis.Methods: There were three groups, the normal group, the model group and the curcumin group.We made adjuvant arthritis rat models with Freund′s complete adjuvant,and they were divided into the model group and the curcumin group.On d 28 after modeling,all rats were killed.All synovial membranes were sent for routine histological evaluation with HE staining.And synovial RANKL and OPG protein expression were measured by Western blot.Results: Curcumin could influence synovium pathology change of adjuvant arthritis rats,the pathological picture showed that infiltration of joint synovial cell,pro-liferation of synovial fibroblasts in model group rats were significantly increased,compared with normal group( P<0.01).Pathology of rats synovial tissue in rats of curcumin group were significantly improved, compared with that in rats of model group.Expression of RANKL,OPG protein in synovial tissue of curcumin group,based on pairwise comparison between with model group,the results were statistically significant( P<0.01 ).Synovial RANKL/OPG ratio were significantly lower than model group ( P<0.01 ).Conclusion:Curcumin can improve synovial pathology damage of adjuvant arthritis rats,curcumin may exert a protective effect on arthritis rats by regulating OPG/RANKL signaling pathway.
		                        		
		                        		
		                        		
		                        	
            

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