1.Sub-chronic aluminum exposure induces cognitive impairment through ALKBH5/PTEN/AKT signaling pathway in rats
Yan LIU ; Xiaoyu ZHANG ; Weitao ZHANG ; Feifan XIAO ; Ping CUI ; Binhong WANG ; Xinru CHEN ; Bin JIANG ; Huan CHEN ; Li LIN ; Jing ZHANG ; Huan LI
China Occupational Medicine 2024;51(2):144-149
ObjectiveTo investigate the effects of maltol aluminum exposure on miR-193a-3p, demethylase AlkB homolog 5 (ALKBH5), phosphatase and tensin homolog deleted on chromosome ten (PTEN) and protein kinase B (AKT), and whether miR-193a-3p is involved in aluminum-induced cognitive impairment by regulating ALKBH5/PTEN/AKT signaling pathway. Methods Specific pathogen-free male SD rats were randomly divided into control group and low-, medium- and high- dose groups according to their body weight, with eight rats in each group. Rats in the low-, medium-, and high- dose groups were intraperitoneally injected with maltol aluminum solution at concentrations of 10.00, 20.00, and 40.00 μmol/kg body weight, respectively, while the rats in control group were given an equal volume of 0.9% sodium chloride solution. Rats were injected for five days every week for three months. After injection, the novel object recognized test was used to assess the learning and memory ability of the rats. The relative expression of miR-193a-3p and B-cell lymphocytoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cysteine aspartate protease-3 (Caspase-3) mRNA in rat hippocampus was detected using the real-time quantitative polymerase chain reaction. The relative protein expression of ALKBH5, PTEN, and AKT2 in the rat hippocampus was detected using Western blot. Results The discrimination index and the preference index of the new object recognition test of the rats in high-dose group were lower than those in control group and low-dose group (all P<0.05). The relative expression of miR-193a-3p and Bcl-2 mRNA in the hippocampus of the rats in high-dose group was lower than those in control group and low-dose group (all P<0.05). The relative mRNA expression of Bax in the high-dose group was higher than those in the control group and low-dose group (both P<0.05). The relative mRNA expression of Caspase-3 of the rats in the high-dose group was higher than that in the other three groups (both P<0.05). The relative protein expression of ALKBH5 in the hippocampus of the rats in the high-dose group was lower than that in the control group (P<0.05). The relative expression of PTEN protein was higher than those in the control group and low-dose group (both P<0.05). The relative protein expression of AKT2 was lower than those in the control group and low-dose group (both P<0.05). Conclusion Sub-chronic aluminum exposure can inhibit the expression of miR-193a-3p in the hippocampus of rats, which may disrupt the ALKBH5/PTEN/AKT pathway and affect normal neuronal homeostasis and cellular function. This pathway may play an important role in aluminum-induced cognitive impairment.
2.The immediate brain effect of acupuncture at Fengchi in patients with posterior circulation ischemia vertigo:a resting-state functional MRI study
Xiaojia YUAN ; Yu XU ; Weitao WANG ; Liqun ZHONG ; Xiaozhen LI ; Dan XU ; Wenlong SHE ; Zhengguang CHEN
Journal of Practical Radiology 2024;40(2):186-189
Objective To investigate the immediate brain effect of acupuncture at Fengchi using amplitude of low-frequency fluctuation(ALFF)and functional connectivity by the resting-state functional magnetic resonance imaging(rs-fMRI)in patients with posterior circulation ischemia vertigo(PCIV).Methods Twenty patients with PCIV were enrolled.The dizziness handicap inventory(DHI)was used to evaluate the severity of vertigo.The patients were randomly divided into acupuncture group and sham acupoint acupuncture group.Rs-fMRI scan was performed before and after acupuncture.MATLAB-based DPABI 6.1 software was used to analyze rs-fMRI data.Correlation analysis was used between the altered ALFF values and DHI scores.The regions of altered ALFF were taken as seeds to analyze functional connectivity.Results Compared with the sham acupoint acupuncture group,the increased ALFF values were mainly located on the left precuneus,left superior frontal gyrus and left caudate nucleus after acupuncture in the acupuncture group;the decreased ALFF values were mainly located on the left cerebellum and right inferior occipital gyrus.The ALFF value of the left superior frontal gyrus was negatively correlated with the DHI score(P=0.04).The increased functional connectivity was mainly found between left precuneus and the right middle frontal gyrus,the right superior frontal gyrus,the decreased functional connectivity was mainly found between left precuneus and the bilateral paracentral lobule and right cerebellum.Conclusion The ALFF value and functional connectivity are different before and after acupuncture,indicating that the vestibular network,visual and motor brain regions functional activities are changed after needling at Fengchi,which may be the brain functional basis of Fengchi for vertigo in PCIV.
3.Prediction of risks of early anastomotic recurrence following primary bowel resection in patients with Crohn's disease based on preoperative magnetic resonance enterography
Weitao HE ; Xiaodi SHEN ; Yangdi WANG ; Jinfang DU ; Xuehua LI ; Shanshan XIONG ; Zhoulei LI ; Shaochun LIN
The Journal of Practical Medicine 2024;40(5):664-671
Objective To develop a nomogram for predicting the risks of early anastomotic recurrence(EAR)after primary bowel resection in patients with Crohn's disease(CD).Methods The patients with CD under-going preoperative magnetic resonance enterography(MRE)and primary bowel resection were enrolled in this retrospective study and divided into an EAR group(18 patients)and EAR-free group(12 patients).The EAR group included the patients having an endoscopic Rutgeerts score of≥I2 month or the need for anastomotic resection within 12 months after surgery.All the 38 indexes including preoperative demographic characteristics,laboratory examina-tions,multi-parameter MRE features of the resected intestine and its adjacent mesentery,histological findings,and postoperative pharmacotherapy were analyzed.Least absolute shrinkage and selection operator(LASSO)regression and multivariate binary logistic regression analysis were performed to identify independent risk factors to be incorpo-rated into the nomogram for predicting the risks of early anastomotic recurrence and the prediction performance was evaluated.Results Mesenteric creeping fat index on MRE and comb sign were independent risks of EAR,with a concordance index of 0.882(95%CI:0.764~1).The calibration plot revealed a strong relationship between actual observation and predicted probability of EAR.Conclusions The preoperative MRE-based nomogram may be a potential tool for predicting EAR following surgery in patients with CD,which is beneficial to individual management in those patients.It provides reference for the formulation of early postoperative individualized drug adjuvant therapy in patients at high risk of EAR.
4.Effect of comprehensive nutrition management on blood glucose and pregnancy outcome of individuals with gestational diabetes mellitus
Rui WANG ; Mingming QI ; Weitao YANG ; Jian HUANG ; Jinyan XIAO ; Yichun LI ; Yonghong WANG ; Yanping LIU
Basic & Clinical Medicine 2024;44(4):434-439
Objective To investigate the effects of comprehensive nutrition management on glycolipid metabolism and pregnancy outcomes in patients with gestational diabetes mellitus(GDM).Methods A total of 121 pregnant women with GDM at 24-28 weeks gestation who were registered in the obstetrics department of 6 sub-central hospi-tals in China from May 2021 to July 2021 were included in this study and were randomly divided into intervention group(n=74)and control group(n=47).The intervention group received intensive comprehensive nutrition man-agement,including at least 6 outpatient interventions,individualized nutrition management and a half-day standard-ized outpatient education on gestational diabetes mellitus,continuous dynamic blood glucose monitoring and micro-blood glucose monitoring,and routine check of glycated albumin and urine every 4 weeks.Body weight,body com-position and diet and exercise implementation procedures and fetal development as well as complications were recor-ded.The control group received conventional nutritional guidance.The two groups were compared for difference in blood glucose related indicators at 37 weeks of gestation,weight gain before delivery,some lipid metabolism indica-tors,pregnancy outcomes,and oral glucose tolerance test(OGTT)at 42 days postpartum.Results Compared with the control group,the level of prenatal fasting blood glucose(P=0.006),intravenous plasma glucose(P=0.009)and blood ketone(P = 0.044)in the intervention group was significantly reduced.There was no significant difference in weight gain and weight attainment rate between the two groups.The 2-hour postpartum OGTTs of preg-nant women in the intervention group(P=0.006)were significantly lower than those in the control group,and the incidence of preeclampsia and postpartum blood loss were lower than those in the control group but no statistical difference was found.For newborns,the incidence of macrosomia(P=0.042)and planation(P=0.048)in the in-tervention group was slightly lower than that in the control group,and the results were statistically different.Other adverse pregnancy outcomes were not statistically different between the two groups.Conclusions Intensive compre-hensive nutrition management has a positive impact on the control of the blood glucose in pregnant women and im-proves the maternal and neonatal outcomes of women with GDM.
5.Effect of miR-192 on the proliferation,migration,and invasion ability of colorectal cancer cell lines
Ying CHEN ; Weitao LI ; Na LI ; Chan ZHOU ; Yongqiang XU
International Journal of Laboratory Medicine 2024;45(1):60-63
Objective To study the effect of microRNA-192(miR-192)on the proliferation,migration and invasion ability of colorectal cancer(CC)cell lines.Methods Group A(SW1116 CC transfected with physio-logical saline),group B(SW1116 CC transfected with miR-192 mimics)and group C(SW1116 CC transfected with miR-192 inhibitor)were set up,respectively.Cell proliferation was detected by MTT assay,cell apoptosis was detected by flow cytometry,cell migration ability was detected by scratch assay,and cell invasion ability was detected by Transwell assay.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression levels of miR-192 and WNT family member 2B(WNT2B)in each group.Results The survival rate and monoclonal number of SW1116 CC cells in group B were(57.32± 6.19)%and(284.59±15.08),which were lower than(76.21±8.23)%and(601.47±23.16)in group A and(89.52±10.62)%and(2 150.68±34.79)in group C,while the apoptosis rate in group B was(20.52± 2.52)%,which was higher than(13.78±1.62)%in group A and(11.62±1.41)%in group C.The survival rate and number of monoclonal formation of SW1116 CC cells in group C were higher than those in group A,while the apoptosis rate was lower than that in group A(all P<0.05).The scratch width of SW1116 CC cells in group B was(785.10±46.18)mm,which was higher than(601.32±33.21)mm in group A and(326.99± 17.48)mm in group C,while the scratch width in group C was lower than that in group A.The number of per-forating cells in group B was(624.67±19.05),which was lower than(875.23±27.30)in group A and(1 204.17±34.59)in group C,and the number of perforating cells in group C was higher than that in group A(all P<0.05).The relative expression level of miR-192 mRNA in SW1116 CC cells in group B was(3.01± 0.26),which was higher than(1.87±0.20)in group A and(0.97±0.23)in group C,and the relative expres-sion level of miR-192 mRNA in group C was lower than that in group A.The expression level of WNT2B mR-NA in group B was(2.16±0.26),which was lower than(4.11±0.50)in group A and(6.08±0.72)in group C,and the expression level of WNT2B mRNA in group C was higher than that in group A(all P<0.05).Con-clusion miR-192 could inhibit the malignant evolution of CC,and one of its main mechanisms may be related to the regulation of WNT2B expression.
6.Causal relationship between sleep phenotype and idiopathic normal pressure hydrocephalus:a two-sample bidirectional Mendelian randomization study
Weitao ZHONG ; Weisong LI ; Zelin LI ; Qiang WANG ; Wangming ZHANG
Journal of Southern Medical University 2024;44(8):1612-1619
Objective To explore the causal relationship between sleep phenotype and idiopathic normal pressure hydrocephalus(iNPH)using two-sample bidirectional Mendelian randomization.Methods The exposure data including 8 sleep phenotypes used in this study were obtained from GWAS catalog,FinnGenR10 and MRCIEU GWAS.The outcome data for idiopathic normal-pressure hydrocephalus were obtained from FinnGen R10.We used the inverse-variance weighted(IVW)method to perform the principal analyses.Cochrane Q-statistics test was used to assess the heterogeneity and MR Egger-intercept test performed to evaluate the pleiotropy for sensitivity analyses.Results IVW result showed that frequent daytime nap was associated with higher odds of iNPH(OR=3.3393,95 CI%:1.0646-10.4742,P=0.0270).Cochrane Q-statistics test and MR Egger-intercept test showed that the MR analysis had no pleiotropy or heterogeneity(P>0.05).The external validation reproduced this result(OR=2.5660,95 CI%:1.1680-5.6373,P=0.0189;OR=4.0424,95 CI%:1.5709-10.4024,P=0.0038).Reverse Mendelian randomization suggested that iNPH did not have significant impact on sleep phenotype.Conclusion The frequency of daytime naps is causally associated with iNPH,and reducing the frequency of weekly daytime naps can reduce the risk of iNPH in the elderly population.
7.Effect of miR-761 on epithelial-mesenchymal transition in osteosarcoma MG63 cells by regulating tumor-associated macrophage polarization
Shilei GAO ; Jiaqiang WANG ; Weitao YAO ; Zhichao TIAN ; Chao LI ; Xiaoxiao LIANG ; Xin WANG
Journal of Jilin University(Medicine Edition) 2024;50(4):978-988
Objective:To discuss the effect of exosome(Exo)microRNA-761(miR-761)on the epithelial-mesenchymal transition(EMT)process of the osteosarcoma(OS)cells by regulating tumor-associated macrophage(TAM)polarization,and to clarify its related mechanism.Methods:The miR-761 plasmid and negative control(miR-NC)plasmid were transfected into the HEK293 cells,and the non-transfected cells were regarded as control group.The transfection efficiency was detected using real-time fluorescence quantitative PCR(RT-qPCR)method.The Exo containing miR-761 was isolated,and the morphology of Exo was observed by transmission electron microscope.The concentration and size distribution of Exo samples were detected by nanoparticle analyzer,and the expression of Exo surface marker protein was detected by Western blotting method.The human monocyte leukemia THP-1 cells were stimulated with phorbol 12-myristate 13-acetate(PMA)to become the M0 macrophages,which were then treated with Exo containing miR-761 and co-cultured with the OS MG63 cells to establish the co-culture system.The experiment was divided into M0 group,TAM group,miR-761 NC group,and miR-761 Exo group.The M0 macrophages were collected from various groups,and the positive rates of M1 macrophage marker CD86 and M2 macrophage marker CD206 in various groups were detected by flow cytometry;the protein expression levels of M1 macrophage secreted factors interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)and M2 macrophage secreted factors interleukin-10(IL-10)and transforming growth factor-β1(TGF-β1)in various groups were detected by Western blotting method.The M0 macrophages were treated with Exo containing miR-761 and co-cultured with MG63 cells to establish the co-culture system.The experiment was divided into control group,TAM group,miR-NC Exo+TAM group,and miR-761 Exo+TAM group.The MG63 cells in various groups were collected,and the fluorescence intensities of E-cadherin and Vimentin in the MG63 cells in various groups were observed by immunofluorescence staining;the expression levels of E-cadherin,Vimentin,and EMT regulation-related transcription factors Twist1,Snail,and Slug proteins in the cells in various groups were detected by Western blotting method;the numbers of invasion and migration cells in various groups were detected by Transwell chamber assay.Results:The HEK293 cells containing miR-761 were successfully obtained by transfection experiments,and the Exo was isolated.Compared with M0 group,the positive rate of CD86 of the macrophages in TAM group was decreased(P<0.05),while the positive rate of CD206 was increased(P<0.05),the expression levels of IL-1β and TNF-α proteins were decreased(P<0.05),while the expression levels of IL-10 and TGF-β1 proteins were increased(P<0.05).Compared with TAM group,the positive rate of CD86 of the macrophages in miR-761 Exo group was increased(P<0.05),while the positive rate of CD206 was decreased(P<0.05),the expression levels of IL-1β and TNF-α proteins were increased(P<0.05),while the expression levels of IL-10 and TGF-β1 proteins were decreased(P<0.05).Compared with control group,the fluorescence intensity of E-cadherin in the MG63 cells in TAM group was decreased,while the fluorescence intensity of Vimentin was increased,the expression level of E-cadherin protein was decreased(P<0.05),while the expression levels of Vimentin,Twist1,Snail,and Slug proteins were increased(P<0.05),and the numbers of invasion and migration cells were increased(P<0.05).Compared with TAM group,the fluorescence intensity of E-cadherin in the MG63 cells in miR-761 Exo+TAM group was increased,while the fluorescence intensity of Vimentin was decreased,the expression level of E-cadherin protein was increased(P<0.05),while the expression levels of Vimentin,Twist1,Snail,and Slug proteins were decreased(P<0.05),and the numbers of invasion and migration cells were decreased(P<0.05).Conclusion:The exo-delivered miR-761 can inhibit the EMT process of the OS cells,thereby inhibiting the cell migration and cell invasion;its mechanism may be related to regulating TAM polarization.
8.Application of peritoneal interposition flap technique in the prevention of pelvic lymphocysts after laparoscopic radical prostatectomy with extended pelvic lymph node dissection
Liqun HUANG ; Xuelei WANG ; Guosheng YANG ; Rongbing LI ; Dongyang LI ; Jing YE ; Weitao HUANG ; Hang WANG ; Jianming GUO ; Xiaofei WEN
Chinese Journal of Urology 2024;45(8):608-613
Objective:To investigate the efficacy of Peritoneal interposition flap (PIF) technique in preventing postoperative pelvic lymphocele formation during laparoscopic radical prostatectomy with extended pelvic lymph node dissection (LRP+ ePLND).Methods:A retrospective analysis was conducted on clinical data of 113 patients with locally high-risk or locally advanced prostate cancer who underwent LRP+ ePLND at Shanghai East Hospital, from January 2020 to November 2023. Among them, 27 patients received PIF technique and 86 received traditional LRP+ ePLND. ePLND was carried out as the clearance of external iliac vessels, medial side of the internal iliac artery, and pararectal lymph nodes. The PIF technique was the suturing the peritoneal flap after freeing the bladder to the lateral side of the bladder, pulling the peritoneal edge that follows the bladder's free edge posteriorly to the pubis, curling it onto the lateral surface of the bladder. This could expose the lymph node clearance bed, establishing a pathway from the lymph node clearance bed to the abdominal cavity space, allowing exuded lymphatic fluid to flow into the abdominal cavity for absorption by the peritoneum. There were no statistically significant differences in age [(68.37±6.92)years vs.(70.47±5.72)years], body mass index [(25.47±2.49)kg/m 2vs.(24.46±2.80)kg/m 2], and preoperative PSA [(23.28±13.94)ng/ml vs.(24.81±13.99)ng/ml] between the PIF group and the control group ( P>0.05). Biopsy Gleason score in PIF group: 6 in 2 cases, 7 in 9 cases, 8 in 9 cases, 9-10 in 2 cases. Biopsy Gleason score in control group: 6 in 4 cases, 7 in 35 cases, 8 in 27 cases, 9-10 in 20 cases. Clinic stage in PIF group: T 2 in 18 cases, T 3 in 6 cases, T 4 in 3 cases. Clinic stage in control group: T 2 in 51cases, T 3 in 27 cases, T 4 in 8 cases. The preoperative Gleason scores and TNM staging comparisons between the PIF group and the control group showed no statistically significant differences ( P>0.05). Surgical duration, intraoperative blood loss, lymph node positivity rate, incidence of postoperative lymphocele, and recovery of urinary control were compared between the two groups. Results:All surgeries were completed successfully without intraoperative complications in both groups. There were no statistically significant differences between the PIF group and the control group in terms of surgical duration [(202.96±24.15)min vs.(201.1±29.85)min], intraoperative blood loss [(85.56±32.27)ml vs.(90.7±49.25)ml], and lymph node positivity rate [(4 in PIF group, 14.8%)vs.(25 in control group, 29.1%)]( P>0.05). Urinary catheters were retained for 10-14 days postoperatively. Following catheter removal, there were no statistically significant differences in urinary control rates at 1 month [51.85%(14/27)vs. 48.83%(42/86)]and 2 months[74.07%(20/27) vs. 72.09%(62/86)] between the PIF group and the control group ( P>0.05). At the 2 to 6-month follow-up CT scan, none of the 27 patients in the PIF group developed pelvic lymphocele, whereas 9 patients in the control group did (6 cases bilateral, 3 cases unilateral), showing a statistically significant difference between the two groups ( P=0.002). Postoperatively, 3 patients in the control group experienced symptoms, with 1 case of lymphocele infection causing fever 1 month after surgery. Lymphocysts were found in 2 patients with ipsilateral lower extremity swelling 2 weeks after surgery. Conclusions:The application of PIF technique during laparoscopic radical prostatectomy with extended pelvic lymph node dissection via the abdominal approach could be safe and feasible. It may prevent postoperative pelvic lymphocele formation.
9.Exploration of pathogenesis and treatment of epilepsy based on astrocyte energy metabolism
Zihao ZHANG ; Yiming LI ; Weitao WANG ; Daopei ZHANG ; Huailiang ZHANG
Chinese Journal of Neuromedicine 2024;23(10):1069-1073
Astrocytes play a key role in maintaining the homeostasis and function of the central nervous system. Abnormal lipid accumulation, ferroptosis, elevated lactic acid, and high DNA methylation mediated by astrocytes in their own or toxic states are particularly closely related to seizures. In recent years, epilepsy treatment from astrocyte energy metabolism perspective has become a research hotspot. Metabolic therapy, such as ketogenic diet, can inhibit seizures via a variety of mechanisms as astrocyte homeostasis maintenance. This paper reviews the relationship between astrocyte energy metabolism and epilepsy pathogenesis and its related metabolic therapy, in order to provide new ideas for clinical treatment of drug-resistant epilepsy.
10.Efficacy of autofluorescence point-spectral analysis combined with the immune colloidal gold technique for the detection of ectopic microscopic parathyroid glands to guide surgery for secondary hyperparathyroidism
Kun PENG ; Baozhong YAO ; Hongcun CHEN ; Jun ZHANG ; Wenzhong BAO ; Wenbo LI ; Weitao SONG ; Sailong SANG ; Li LIN ; Zhixing JIA ; Liang LI
The Journal of Practical Medicine 2024;40(20):2905-2912
Objective To evaluate the intraoperative identification of ectopic parathyroid tissue in the central neck region using autofluorescence point-spectral analysis(AFPSA)combined with immune colloidal gold technique(ICGT),for guiding total parathyroidectomy(TPTX)or clean parathyroidectomy(CPTX)in the management of secondary hyperparathyroidism(SHPT).Methods Retrospectively collected and compared the clinical data of 64 patients with SHPT from October 2019 to June 2023.In the observation group,TPTX was performed as the initial procedure in 36 cases,followed by sampling of suspicious targets using AFPSA in the central neck area and subsequent detection through ICGT.CPTX was then conducted if a positive result was obtained.On the other hand,the control group consisted of 28 cases where only TPTX was performed without any additional tests during surgery.The surgical data,parathyroid hormone(PTH)levels,blood calcium levels,blood phosphorus levels,alkaline phosphatase(ALP)levels,regression of clinical symptoms,changes in parathyroid function and occurrence of hypocalcemia were compared between these two groups.Results In the observation group,there were 9 cases of AFPSA-ICGT positivity,including 2 left-sided cases,4 right-sided cases,and 3 thymic cases;among these posi-tive cases,there were a total of 10 locations with mildly hyperplastic or nonhyperplastic microscopic parathyroid tissue.The difference in the number of total parathyroid glands removed(including ectopic)between the two groups was statistically significant(P<0.05).At both 3 and 6 months postoperatively,ALP levels in the observation group were significantly lower than those in the control group(P<0.01 and P<0.001 respectively);at 6 months postoperatively,differences in PTH and blood phosphorus levels between the two groups were also statistically significant(P<0.05 and P<0.001 respectively).Joint bone pain and skin itching recurred in some patients within the control group at six months after surgery(P<0.05),whereas recurrence of SHPT was less frequent within the observation group compared to controls(P<0.05);however,no statistically significant differences were observed regarding postoperative hypoparathyroidism or hyperparathyroidism as well as hypocalcemia between either groups.Conclusion The AFPSA-ICGT intraoperative test can be utilized to guide surgery for SHPT,enabling accurate and efficient identification as well as safe targeting of parathyroid tissues that may not exhibit obvious hyperplasia in the central cervical region.

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