1.Increased expression of autoantibody protein and soluble IL-6 receptor in serum of patients with Tourette's syndrome
Yuhang CHENG ; Yi ZHENG ; Lifang WANG ; Ruimei LIU ; Yang GAO
Basic & Clinical Medicine 2010;30(2):113-116
Objective To explore the mechanism of immunologic injury in patients with Tourette's syndrome(TS). Methods The serum level of anti-brain antibody ( ABAb) , antinuclear antibody ( ANAb) , soluble IL-6 receptor (sIL-6R) and soluble gpl30(sgp130) in patients with TS were analyzed by commercially available ELISA kits,and the titer of anti-streptolysin O ( ASO) in TS and in control was examined with latex enhanced immunoturbidimetry. Results The level of sIL-6R and sgpl30 was significantly elevated in TS compared with control group [(44. 1 ± 15.8)ng/mL vs (30. 3 ± 9. 0) ng/mL and (69. 0 ±24. 6)ng/mL vs (47. 3 ±14. l)ng/mL,P <0. 01 respectively]. ASO titer(250 U/mL) was found higher in TS than that in control(P <0. 01). The positive rates of anti-brain an-tibody and antinuclear antibody in TS were higher than that in control group (66% vs 4% and 53% vs 25% , P <0. 01 respectively). The level of ABAb negatively correlated with sgpl30 concentration( r = 0. 375, P <0. 05 ). Conclusion IL-6 signal transduction might be involved in Tourette's syndrome to lead the function enhancementand start cascade of feedback inhibition, because of elevated levels of slL-6R, sgp130, ANAb and ABAb in serum.Autoimmune-related injuries may potentially explain the pathogenesis of the disease.
2.Role of MafA gene in insulin production-Analysis of heterozygous knockout mice
Chuan ZHANG ; Bo LI ; Yan CHENG ; Yuhang YAO ; Lijuan SUN ; Satoru TAKAHASHI
Chinese Journal of Endocrinology and Metabolism 2013;29(12):1048-1051
Objective To clarify the role of MafA gene in development of MODY (maturity onset diabetes of the young) by studying insulin production,gene expression,and serum glucose level in heterozygous MafA gene knockout mice.Methods C57BL/6J mice were used as control animals,MafA gene heterozygous mice were analyzed.The distribution curve of blood sugar levels over time and serum insulin of heterozygous mice were determined by using IPGTT.The sensitivity of the mice to insulin was examined by injecting insulin assay.The expression levels of genes of MafA,insulin,pdX1,Beta2,and other genes of heterozygous mice were analyzed by semi-quantitative RT-PCR.Morphological changes in pancreatic tissue and α-and β-cell counts were obtained by using immunofluorescence/histological examination.Results (1) Two weeks after birth,MafA gene heterozygous mice began to show that the blood glucose level was increased,weight was reduced,and the amount of insulin secretion was clearly decreased (P<0.05 or P<0.01) while the insulin sensitivity did not change significantly.(2)The islet volume in MafA gene heterozygous mice was increased significantly as compared with the control group.However there were no significant changes in the number of pancreatic cells,distribution patterns,and the ratio of α and β cell.(3) Semi-quantitative RT-PCR detection showed that,compared with the control group,MafA gene level,the amount of insulin and Beta2 gene in MafA gene heterozygous mice were significantly reduced(all P<0.05),while no changes in the amount of glucagons and level of Pdx1 were found.Conclusions The blood glucose level of MafA gene heterozygous mice was raised early after birth.MafA gene is likely to be a new disease ralated gene of MODY.
4.Inhibitory effect of siRNA-KCNQ1OT1 on human lens epithelial cell apoptosis by targeting miR-199a-5p
Cheng LU ; Fengyan ZHANG ; Yuhang ZHANG ; Jiajuan ZHANG
Chinese Journal of Experimental Ophthalmology 2022;40(6):514-523
Objective:To explore the inhibitory effect of long non-coding RNA (lncRNA) KCNQ1 overlapping transcript 1 (KCNQ1OT1) by targeting microRNA-199a-5p (miR-199a-5p) on the apoptosis of human lens epithelial cells (LECs).Methods:The anterior lens capsule tissue of 23 age-related cataract patients who underwent cataract surgery in Xinxiang First People's Hospital from December 2018 to August 2019 was collected.At the same time, anterior lens capsules from 20 healthy donor were collected.The expressions of KCNQ1OT1 and miR-199a-5p in the tissues were detected by real-time fluorescence PCR.Human LECs SRA01/04 cultured in vitro were divided into blank control group, model control group, small interfering RNA-negative control (siR-NC) group, siR-KCNQ1OT1 group, miR-NC group, miR-199a-5p group, siR-KCNQ1OT1+ anti-miR-NC group and siR-KCNQ1OT1+ anti-miR-199a-5p group.No intervention was administered to blank control group.Cells in model control group were cultured with 100 μmol/L H 2O 2 for 24 hours to establish oxidative stress injured model, and cells in the other six groups were transfected with corresponding transfection reagents for 6 hours by liposome method according to grouping, and then treated with 100 μmol/L H 2O 2 for 24 hours.The expressions of KCNQ1OT1 and miR-199a-5p in lens anterior capsule tissue and LECs cells were determined by real-time fluorescent quantitative PCR.Cell viability was detected with thiazolyl blue (MTT). Cell apoptosis was analyzed by flow cytometry.The expressions of B-cell lymphoma/leukemia-2 (bcl-2) and bcl-2 related X protein (Bax) proteins were assayed by Western blot.The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured by enzyme-linked immunosorbent assay (ELISA). The targeting relationship between KCNQ1OT1 and miR-199a-5p was verified by dual luciferase reporter experiment.The study protocol was approved by an Ethics Committee of Xinxiang First People's Hospital (No.2019-001). Written informed consent was obtained from relatives of patient. Results:The relative expression of KCNQ1OT1 in the anterior capsule of patients with age-related cataract was 2.41±0.42, which was significantly higher than 0.97±0.19 of normal people, and the relative expression of miR-199a-5p in the capsule of patients with age-related cataract was 0.36±0.12, which was lower than 1.04±0.15 of normal people, and the differences were statistically significant ( t=14.112, 16.507; both at P<0.001). Compared with blank control group, the relative expressions of KCNQ1OT1 and bax protein, cell apoptosis rate and MDA content were significantly increased, and the relative expressions of miR-199a-5p and bcl-2 protein, cell viability and SOD activity were significantly reduced in model control group, showing statistically significant differences (all at P<0.001). Compared with siR-NC group, the relative expressions of KCNQ1OT1 and bax protein, cell apoptosis rate and MDA content in cells of siR-KCNQ1OT1 group were decreased, while the relative expression of bcl-2 protein, cell survival rate and SOD activity were increased, and the differences were statistically significant (all at P<0.05). Compared with miR-NC group, the KCNQ1OT1-wild type (WT) luciferase activity in miR-199a-5p group was significantly decreased, with a statistically significant difference ( t=21.131, P<0.001). The relative expression levels of miR-199a-5p and bcl-2 proteins, cell survival rate and SOD activity were significantly increased, and the relative expression of bax protein, cell apoptosis rate and MDA content were significantly decreased in miR-199a-5p group than those in miR-NC group, and the differences were statistically significant (all at P<0.05). The relative expression levels of miR-199a-5p and bcl-2 proteins, cell survival rate and SOD activity were significantly lower, and the cell apoptosis rate, relative expression of bax protein and MDA content were significantly higher in siR-KCNQ1OT1+ anti-miR-199a-5p group than those in siR-KCNQ1OT1+ anti-miR-NC group, and the differences were statistically significant (all at P<0.05). Conclusions:The inhibition of KCNQ1OT1 can promote the cell viability of human LECs, inhibit H 2O 2-induced cell apoptosis and oxidative stress, and the mechanism may be related to the up-regulation of miR-199a-5p.
5.Analysis on detection situation of motor cognitive risk syndrome and risk factor
Lang PENG ; Honghan LIU ; Jingling CAI ; Shichao ZHAO ; Yuhang CHENG ; Zhengliang QI ; Qingwei XIANG
Chinese Journal of Geriatrics 2022;41(3):261-265
Objective:To investigate the detection rate of motoric cognitive risk(MCR)syndrome and explore the possible risk factors at different age groups.Methods:A total of 561 patients from geriatric outpatient clinic of Hubei Provincial Hospital of Traditional Chinese Medicine from November 2018 to December 2019 were divided into two age groups under 70 years old(n=241)and 70 years old and above(n=320). The general information, Pittsburgh Sleep Quality Index, Geriatric Depression Scale-15(GDS-15), 4-meter walking test, Mini-Mental State Examination and Morse Fall Scale were collected.Patients with MCR were screened out according to the MCR diagnostic criteria.Logistic multiple regression analysis was used to analyze the associated risk factors.Results:7 cases(7/241, 2.9%)met the MCR diagnostic criteria in age<70 years group, and 34 cases(34/320, 10.7%)in age ≥ 70 years group.The proportion of hearing impairment complaints and GDS-15 scores of MCR patients were higher than those of the non-MCR group in age<70 years group, and the Morse Fall Scale of MCR patients was higher than that of the non-MCR group in age ≥70 years old group( P<0.05). After adjusting for associated confounding factors, multiple logistic regression analysis showed that hearing impairment complaints( OR=26.394, P<0.05)and GDS-15( OR=1.385, P<0.05)were independent risk factors for MCR in age<70 years group.And female( OR=0.445, P<0.05)was a protective factor for MCR in age ≥70 years old group. Conclusions:Motoric cognitive risk syndrome has different risk factors in different age groups, which may indicate that the causes and predictive significance of MCR in these two different age groups are different.
6.Comparison of ultrasound-guided transrectal and transperineal prostate biopsies in clinical application.
Li-rong YUAN ; Cheng-guang ZHANG ; Lai-xing LU ; Lei RUAN ; Jian-hong LAN ; Sen-qiang FENG ; Jin-dan LUO
National Journal of Andrology 2014;20(11):1004-1007
OBJECTIVETo compare the positive rates and complications of ultrasound-guided transrectal and transperineal prostate biopsies.
METHODSWe retrospectively analyzed 156 cases of ultrasound-guided transrectal (n = 97) and transperineal (n = 59) prostate biopsy, and compared the positive rate and post-biopsy complications between the two approaches.
RESULTSThe positive rates in the transrectal and transperineal groups were 48.4% and 44.1%, respectively, with no significant difference between the two approaches according to different PSA levels (P >0.05). No statistically significant differences were observed between the transrectal and transperineal groups in the post-biopsy incidence rates of such complications as hematuria (54.6% vs 42.4%, P >0.05), lower urinary tract symptoms (17.5% vs 22.0%, P >0.05), dysuria (9.3% vs 6.8%, P >0.05), and acute urinary retention (7.2% vs 6.8%, P >0.05). However, the incidence rates of post-biopsy infection and rectal bleeding were remarkably higher (15.5% vs 3.4%, P<0.05 and 50.5% vs 3.4%, P >0.01) while that of perineal swelling markedly lower in the former than in the latter (3.1% vs 13.6%, P <0.05).
CONCLUSIONTransrectal and transperineal biopsies are both effective for the diagnosis of prostate cancer. Since their complications vary, the choice between the two methods depends on the specific condition of the patient.
Biopsy, Needle ; adverse effects ; methods ; Hematuria ; etiology ; Humans ; Lower Urinary Tract Symptoms ; etiology ; Male ; Prostate ; pathology ; Prostatic Neoplasms ; pathology ; Rectum ; Retrospective Studies ; Ultrasonography, Interventional ; methods ; Urination Disorders ; etiology
7.MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus Cavernosum by Targeting Krüppel-Like Factor 4
Daoyuan HU ; Yunlong GE ; Yuhang XI ; Jialiang CHEN ; Hua WANG ; Chi ZHANG ; Yubin CUI ; Lizhao HE ; Ying SU ; Jun CHEN ; Cheng HU ; Hengjun XIAO
The World Journal of Men's Health 2024;42(3):638-649
Purpose:
The poor retention and ambiguous differentiation of stem cells (SCs) within corpus cavernosum (CC) limit the cell application in erectile dysfunction (ED). Herein, the effects and mechanism of microRNA-145 (miR-145) gene modification on modulating the traits and fate of bone marrow-derived mesenchymal stem cells (BMSCs) were investigated.
Materials and Methods:
The effects of miR-145 on cell apoptosis, proliferation, migration, and differentiation were determined by flow cytometry, cell counting kit-8, transwell assays and myogenic induction. Then, the age-related ED rats were recruited to four groups including phosphate buffer saline, BMSC, vector-BMSC, overexpressed-miR-145-BMSC groups. After cell transplantation, the CC were harvested and prepared to demonstrate the retention and differentiation of BMSCs by immunofluorescent staining. Then, the target of miR-145 was verified by quantitative real-time polymerase chain reaction and immunohistochemical. After that, APTO-253, as an inducer of Krüppel-like factor 4 (KLF4), was introduced for rescue experiments in corpus cavernosum smooth muscle cells (CCSMCs) under the co-culture system.
Results:
In vitro, miR-145 inhibited the migration and apoptosis of BMSCs and promoted the differentiation of BMSCs into smooth muscle-like cells with stronger contractility. In vivo, the amount of 5-ethynyl-2′-deoxyuridine (EdU)+cells within CC was significantly enhanced and maintained in the miR-145 gene modified BMSC group. The EdU/CD31 co-staning was detected, however, no co-staining of EdU/α-actin was observed. Furthermore, miR-145, which secreted from the gene modified BMSCs, dampened the expression of KLF4. However, the effects of miR-145 on CCSMCs could be rescued by APTO-253.
Conclusions
Overall, miR-145 modification prolongs the retention of the transplanted BMSCs within the CC, and this effect might be attributed to the modulation of the miR-145/KLF4 axis. Consequently, our findings offer a promising and innovative strategy to enhance the local stem cell-based treatments.
8.The preliminary study about feasibility and accuracy of the frozen section during targeted prostate biopsy
Miao WANG ; Zhengtong LYU ; Xuan WANG ; Huimin HOU ; Yuhang FU ; Hao CHENG ; Chunmei LI ; Longteng LIU ; Jia CHEN ; Xin DING ; Jianye WANG ; Ming LIU ; Wei ZHANG
Chinese Journal of Urology 2022;43(7):490-494
Objective:To investigate the feasibility and reliability of the frozen section during targeted prostate biopsy.Methods:The clinical and pathological information of patients who received cognitive fusion transperineal targeted plus systematic biopsy and frozen section of 1-2 core targeted biopsy were consecutively collected and retrospectively studied. The median age was 70 (ranging 64-78) years, with the median prostate-specific antigen (PSA) level of 11.00 (ranging 6.63-16.52) ng/ml and the median prostate volume of 35.72 (ranging 22.59-47.71) ml. All patients received bi-parametric magnetic resonance imaging (bp-MRI) and have Prostate Imaging Reporting and Data System (PI-RADS) 3 or higher lesions diagnosed on bp-MRI. The suspected lesions would be taken by targeted biopsy of which one or two cores would be sent to prepare for the frozen sections. Then a cognitive fusion targeted and systematic biopsy covering the above targeted zones would be routinely administered under a transperineal approach as a standard protocol. The total time used for diagnosis of the frozen sections, the pathological diagnosis and the International Society of Urological Pathology (ISUP) grade groups (GG) would be recorded. The sensitivity, the positive predictive value, and the accuracy on grade groups would be analyzed, using the pathological diagnosis based on standard sections from the same targeted lesion.Results:A total of 29 patients were included in this study. Accordingly, 29 suspected lesions were identified on bp-MRI. A total of 20 lesions were finally diagnosed of PCa on frozen section, with the detection rate of 69.0%. Of those, 9(45.0%) cases were ISUP GG 1 diseases, 5(25.0%) cases were GG 2 diseases, 1(5.0%) case was GG 3 disease, and 5(25.0%) cases were GG 4-5 diseases. A total of 22 lesions were diagnosed with PCa on standard sections of cores from the same targeted lesions, with the detection rate of 75.9%. Of those, 6(27.3%) cases were GG 1 disease, 11(50.0%) cases were GG 2 diseases, 1(4.5) case was GG 3 disease, and 4(18.2%) cases were GG 4-5 diseases. The sensitivity and the positive predictive value of frozen section were 90.9% and 100%, respectively. No false positive diagnosis was made by frozen section. Compared to diagnosis from frozen sections, the GG diagnosed from final standard sections were found to upgrade and downgrade in 2 and 2 cases, respectively. The accuracy rate on GG of frozen sections was 80%. The time used for the diagnosis of frozen sections was (11±2) minutes. The histology quality control of four specimens was dissatisfactory. Two were due to tissue loss and deformation during sampling, and the other two were due to cytoclasis during low-temperature transferring.Conclusion:It is feasible and reliable to make a pathological diagnosis from frozen section of prostate targeted biopsy.
9. Diabetes mellitus concurrent aspergillus pneumonia: One case report
Chuanfeng LIU ; Yue ZHOU ; Yuhang ZHAO ; Bingzi DONG ; Bingfei CHENG ; Fangchao LIU ; Shengnan SUN ; Yangang WANG
Chinese Journal of Endocrinology and Metabolism 2020;36(2):150-152
Diabetic patients with poor glycemic control are exposed to media containing mold spores, and spores enter the body, which may lead to refractory infections. This article combines case and literature reviews, proposes the diagnosis and treatment method of mold infection, and provides some guidances for subjects who long-term exposure to mold groups such as farmers and immunocompromised people.
10.Effects of EFHD2 protein deletion on tight junction proteins in mouse Sertoli cells
Xiaolin Ye ; Guolin Xu ; Tijun Qian ; Feng Qin ; Yuntao Wang ; Yuhang Cheng ; Wenzhen Zhao
Acta Universitatis Medicinalis Anhui 2022;57(11):1744-1749
Objective :
To study the effect of EFHD2 protein deletion in Sertoli cells on Occludin,a component of tight junction protein and the localization and expression of EF-hand domain family member D2 (EFHD2) in mouse testis.
Methods :
Total RNA and protein were extracted from adult mice's heart ,liver ,spleen ,lung ,kidney, brain and testis tissues.The mRNA and protein levels of EFHD2 in each organ tissue were detected by qRT-PCR and Western blot.Immunofluorescence and immunohistochemistry detected the localization and expression of EF- HD2 in testicular tissues.SiRNA interference was used to reduce EFHD2 in Sertoli cells to detect Occludin protein expression.
Results :
qRT-PCR showed that the expression of EFHD2 was the highest in the testis.Western blot results showed that the expression level increased in testis tissue.Indirect immunofluorescence and immunohisto- chemistry results showed that the protein was mainly distributed in Sertoli cells and co-localized with cytoskeletal Vimentin,indicating that the protein was expressed in Sertoli cells.After the decrease of EFHD2 protein expres- sion,Occludin protein expression also decreased.
Conclusion
The expression of EFHD2 protein in the testis is relatively high,mainly distributed in Sertoli cells of the testis,co-localized with Vimentin,and can affect the nor- mal expression of tight junction protein Occludin.It is suggested that EFHD2 can promote and maintain the junction structure of Sertoli cells and provide a stable microenvironment for spermatogenesis.