1.CircRNA_PLEKHM3 regulates epithelial mesenchymal transformation of cervical cancer cells through the miR-320/KLF4 axis
Yanan ZHANG ; Ying CUI ; Tianjiao WANG ; Zhonglei DU
Acta Universitatis Medicinalis Anhui 2024;59(3):403-412
Objective To investigate the role and mechanism of circular RNA containing pleckstrin homology do-main of Pleckstrin homology domain family M member 3(circRNA_PLEKHM3)in regulating epithelial-mesenchy-mal transition(EMT)behavior in cervical cancer cells through the miR-320 and KLF4.Methods The expression levels of circRNA_PLEKHM3 in cervical cancer cells Hela and CaSki were detected by real-time quantitative PCR(qRT-PCR).RNA fluorescence in situ hybridization was used to determine the localization of circRNA_PLEKHM3 in human cervical cancer epithelial cells CaSki.Dual luciferase reporter gene experiments were conducted to inves-tigate the targeting relationship between circRNA_PLEKHM3 and miR-320,as well as the targeting relationship be-tween miR-320 and KLF4.CaSki cells were overexpressed with circRNA_PLEKHM3.Additionally,three groups were set up:overexpression of miR-320 on the basis of circRNA_PLEKHM3 overexpression,silencing of KLF4 on the basis of circRNA_PLEKHM3 overexpression,and silencing of KLF4 on the basis of miR-320 overexpression.qRT-PCR was performed to detect the expression levels of miR-320 in CaSki.Western blot experiments were con-ducted to determine the expression of KLF4 and EMT markers including E-cadherin,N-cadherin,Vimentin,MMP-2,and MMP-9 in CaSki cells.Transwell assays were performed to measure cell migration and invasion.Results The expression of circRNA_PLEKHM3 decreased in Hela and CaSki cells(P<0.05),mainly localized in the cy-toplasm.The dual luciferase reporter gene experiment demonstrated a targeting relationship between miR-320 and circRNA_PLEKHM3,as well as between KLF4 and miR-320.Overexpression of circRNA_PLEKHM3 inhibited the protein expression of miR-320,N-cadherin,Vimentin,MMP-2,and MMP-9,up-regulated the protein expression of E-cadherin,and reduced cell migration and invasion(P<0.05).Overexpression of miR-320 or silencing of KLF4 on the basis of circRNA_PLEKHM3 overexpression both promoted the protein expression of miR-320,N-cad-herin,Vimentin,MMP-2,and MMP-9,down-regulated the protein expression of E-cadherin,and increased cell migration and invasion(P<0.05).However,silencing of KLF4 on the basis of miR-320 overexpression inhibited the protein expression of KLF4,N-cadherin,Vimentin,MMP-2,and MMP-9,up-regulated the protein expression of E-cadherin,and reduced cell migration and invasion(P<0.05).Conclusion Overexpression of circRNA_PLEKHM3 regulates EMT in cervical cancer cells through the miR-320/KLF4 axis.
2.Relationship between plasma mtDNA,tissue MIP1α,MCP-1 and recovery of hip function in patients with hip fractures
Zhonglei SU ; Yuntao GUO ; Jinfeng LIU ; Wenjuan WANG ; Yuantao DUAN ; Xin'an GUO
Journal of Clinical Surgery 2024;32(11):1200-1204
Objective To analyze the relationship between plasma mitochondrial DNA(mtDNA),macrophage inflammatory protein-1α(MIP1α)and monocyte chemotactic protein 1(MCP-1)in vastus lateralis tissues and postoperative muscle atrophy,recovery of hip function in patients with hip fractures.Methods A total of 86 patients with hip fractures and 43 patients with coxitis in Jinan Eighth People's Hospital were enrolled as hip fracture group and coxitis group between October 2020 and October 2022,respectively.The lateral muscle tissues were collected as samples during surgery.The level of plasma mtDNA was detected by real-time fluorescence quantitative polymerase chain reaction.Before surgery,levels of serum interleukin-6(IL-6)and tumor necrosis factor α(TNF-α)were detected by enzyme-linked immunosorbent assay.Before surgery,cross-sectional areas of types Ⅰ and Ⅱ vastus lateralis fibers were detected by immunofluorescence method.Before surgery,expression levels of MIP1α and MCP-1 proteins in lateral muscle tissues were detected by Western blot.All patients with hip fracture were effectively followed up for 6 months after surgery.At 3 and 6 months after surgery,total lean mass(TLM)and unaffected limb lean mass(ULLM)were detected by DXA.Results The level of plasma mtDNA in hip fracture group was higher than that in coxitis group before surgery[(4.12±0.53)vs(2.37±0.36),P<0.05],levels of serum IL-6 and TNF-α were higher than those in coxitis group[(34.68±6.14)pg/ml,(21.54±4.12)pg/ml vs(12.74±3.06)pg/ml,(10.81±2.71)pg/ml,P<0.05],cross-sectional areas of types Ⅰ and Ⅱ vastus lateralis fibers were smaller than those in coxitis group[(4321.45±441.36)μm2,(2384.38±247.11)μm2 vs(5417.63±553.27)μm2,(3569.24±368.22)μm2,P<0.05],and expression levels of MIP1α and MCP-1 proteins were higher than those in coxitis group[(2.34±0.25),(2.47±0.28)vs(1.18±0.15),(1.95±0.23),P<0.05].In patients with hip fracture after 6 months of follow-up,there were 53 cases with good prognosis and 33 cases with poor prognosis.The level of plasma mtDNA in poor prognosis group was higher than that in good prognosis group before surgery[(4.53±0.52)vs(3.87±0.44),P<0.05],levels of serum IL-6 and TNF-α were higher than those in good prognosis group[(35.97±5.32)pg/ml,(20.74±4.27)pg/ml vs(33.51±5.16)pg/ml,(22.83±4.33)pg/ml,P<0.05],cross-sectional areas of types Ⅰ and Ⅱ vastus lateralis fibers were smaller than those in good prognosis group[(4174.26±434.60)μm2,(2309.56±246.18)μm2 vs(4394.42±450.12)μm2,(2430.97±250.72)μm2,P<0.05],and expression levels of MIP1α and MCP-1 proteins were higher than those in good prognosis group[(2.47±0.28),(1.95±0.23)vs(2.26±0.24),(1.82±0.21),P<0.05].TLM and ULLM at 6 months after surgery were lower than those at 3 months after surgery in good prognosis group and poor prognosis group(P<0.05).At 3 and 6 months after surgery,there was no significant different in TLM or ULLM between good prognosis group and poor prognosis group(P>0.05).Conclusion Traumatic stress injury will increase level of plasma mtDNA in patients with hip fracture,which will induce the increase of systemic inflammatory indexes(serum IL-6,TNF-α)and inflammatory factors(MCP-1,MIP1α)levels,aggravate muscle atrophy and cause postoperative decline of hip function.
3.Clinical Efficacy of Gandouling Combined with Short-term Decoppering on Hepatic Fibrosis of Wilson's Disease Based on Transient Elastography
Yu ZHANG ; Meixia WANG ; Jing ZHANG ; Qian ZHAO ; Meng ZHANG ; Yiming CHEN ; Zhonglei LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(13):123-129
ObjectiveTo explore the accuracy of clinical common serum fibrosis indexes hyaluronic acid (HA), type Ⅳ collagen (CⅣ), laminin (LN), and type Ⅲ procollagen peptide (PⅢNP), in combination with liver stiffness measurement (LSM, measured by transient elastography) and non-invasive markers of fibrosis aspartate aminotransferase to platelet ratio index (APRI) and fibrosis-4 (FIB-4) in the prediction of the hepatic fibrosis of Wilson's disease (WD) and to observe the clinical effect of Gandouling (GDL). MethodThe data of 76 WD patients were collected and the LSM, serum fibrosis indexes (HA, PⅢNP, CⅣ, LN), APRI, and FIB-4 before treatment were recorded. The correlation of LSM with serum fibrosis indexes, APRI, and FIB-4 was discussed via Pearson′s correlation analysis. According to the therapeutic schemes, patients were classified into the control group (36 cases) and treatment group (40 cases). Patients in control group were treated with sodium dimercaptopropylsulfonate (DMPS), while those in the treatment group received GDL in addition to the western medicine therapy. The treatment lasted 6 courses (8 days/course) and the influence of GDL on the indictors was evaluated. ResultHA, CⅣ, LN, PⅢNP, APRI, and FIB-4 were in positive correlation with LSM (r=0.517, 0.438, 0.281, 0.457, 0.778, 0.847, P<0.01). HA, CⅣ, LN, and PⅢNP in the treatment group were lower after treatment than before treatment (P<0.05, P<0.01). HA, CⅣ, and LN in the control group were lower after treatment than before treatment (P<0.05, P<0.01), and PⅢNP showed no significant difference. LSM, FIB-4, and APRI in both groups decreased after treatment (P<0.05). After treatment, LSM, FIB-4, APRI, HA, and PⅢNP in the treatment group were lower than those in the control group (P<0.05, P<0.01), but CⅣ and LN demonstrated no significant difference from the control group. ConclusionLSM in combination with serum fibrosis indexes (HA, PⅢNP, CⅣ, LN), FIB-4, and APRI can help accurately identify the level of the hepatic fibrosis in WD. Moreover, on the basis of decoppering by western medicine, GDL can significantly improve the liver function and hepatic fibrosis of WD patients.
4.Application of laparoendoscopic single-site combined with transurethral approach for unilateral retrograde nephroureterectomy
Zhonglei DENG ; Xuelin SU ; Jian SU ; Luming SHEN ; Yang ZHANG ; Lin YUAN ; Ninghong WANG ; Guojiang XU ; Ping ZHOU ; Qingyi ZHU
Chinese Journal of Urology 2020;41(11):820-824
Objective:To investigate the feasibility and safety of laparoendoscopic single-site combined with transurethral approach for unilateral retrograde nephroureterectomy in the treatment of upper urinary tract epithelial carcinoma.Methods:The clinical data of 12 patients from January 2018 to November 2019 with unilateral retrograde nephroureterectomy were analyzed retrospectively. There were 7 males and 5 females with an average age of 65.9 years, the age ranged from 50 to 78 years.There were 8 cases with left ureteral tumor, 6 cases with left renal pelvis tumor, 4 cases with right tumor(2 cases of right ureteral tumor and 2 cases of right renal pelvis tumor). Surgical methods: 1470 laser sleeve was used to remove the inner segment of the ureter bladder wall after the lower ureter was clipped through abdominal approach, and the urethra was inserted under the guidance of zebra guide wire.The operation time, intraoperative blood loss, intraoperative auxiliary cannula, postoperative hospital stay, postoperative drainage tube removal time, intraoperative and postoperative complications, postoperative pathology were recorded.Results:All of the operations were successful. The mean operation time was 194(135-260)min, the mean estimated blood loss was 50(25-100) ml, and the mean hospitalization time was 11.6(5-24)d. Among the 12 patients, 8 patients had abdominal drainage tube after operation. The mean time for drainage was 6.8(3-11)d. One patient added a 5 mm ancillary port.One patient had urinary leakage at the bladder anastomotic site, the catheter was removed 3 weeks later. The other patients had no postoperative incision infection, fever, bleeding, venous thrombosis and other related complications.No patient received blood transfusion and the pathological margin was negative. The median follow-up time was 12 months (5-15 months). One patient died of lumbar metastasis 8 months after operation, and others were neither tumor recurrence nor distant metastasis.Conclusions:The application of laparoendoscopic single-site combined with transurethral approach for unilateral retrograde nephroureterectomy in the treatment of upper urinary tract epithelial carcinoma is safe, accurate and effective, with less trauma and less bleeding. It is worth applying in clinical practice.
5.A comparative study of cognitive functions in individuals at ultra high-risk for psychosis
Zhonglei WANG ; Cailan HOU ; Yi TANG ; Fujun JIA
Chinese Journal of Behavioral Medicine and Brain Science 2015;24(8):706-709
Objective To investigate the differences of cognitive functions among ultra high-risk individuals(UHR),first-episode schizophrenia patients and healthy controls.Methods Twenty four participants meeting clinical inclusion criteria for initial prodromal states (IPS) selected from the first degree relatives of schizophrenia,23 first onset schizophrenia patients from the same family with UHR as well as 30 healthy controls were assessed with a comprehensive neuropsychological test battery including Trail Making Test (TMT),Stroop color word test and the Continuous Performance Test (CPT) to test the speed of information processing and sustained attention ability.Results ①The time of the Trail Making Test in UHR ((45.12±14.87) s) was shorter than first-episode schizophrenia patients((60.43± 19.30)s),but longer than healthy controls((27.13±7.74) s).The whole score of the Stroop color word test in UHR (28.46± 7.87) was higher than first-episode schizophrenia patients (18.78± 6.86),and lower than healthy controls (43.40±9.64).The score of the Continuous Performance Test in UHR (1.86 ± ± 1.05) was significanly higher than first-episode schizophrenia patients(1.17±0.67),but lower than healthy controls (2.63±0.67),and all these differences were significant (P<0.01).②Taking age,years of education,occupation status,marital status and the Positive and Negative Syndrome Scale (PANSS) scores as the covariance,the analysis of covariance showed TMT and Stroop color word test were still statistically significant.Conclusion There are cognitive function changes in individuals at ultra high-risk for psychosis,lying between first-episode schizophrenia cases and healthy controls.
6.Optimization on Hydrolysis Conditions of Coix Prolamin and Angiotensin-converting Enzyme Inhibitory Activity of Hydrolysates
Jianna YUAN ; Xiumei CHEN ; Shuai CUI ; Yingxi LIANG ; Zhonglei WANG ; Xiaohua ZHANG ; Lingzhi WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(4):806-810
This study was aimed to optimize the hydrolysis conditions of Coix prolamin, which occupied the highest content in total protein of it, and then evaluate the angiotensin-converting enzyme (ACE) inhibitory activity of the corresponding hydrolysates. Pepsin was used to hydrolyze prolamin and OPA method was applied to determine the degree of hydrolysis. The optimum conditions of hydrolyzing prolamin were obtained by orthogonal design subse-quently. The ACE inhibitory activity of small molecular weight (less than 3 kD) peptides gained by ultrafiltration of the hydrolysates was assayed by RP-HPLC method. The result showed that the optimized hydrolysis conditions were substrate concentration of 1%, enzyme-to-substrate ratio of 1:5 and hydrolysis duration for 48 h. The hydrolysates exhibited an ACE inhibitory ratio of (83.40 ± 0.93)% at the concentration of 0.1 mg·mL-1. It was concluded that prolamin hydrolysates displayed a high ACE inhibitory activity in vitro, which provided guidance for further research of antihypertensive component and development of functional food from Coix.
7.Optimization of Preparation Process for 5, 7, 3', 4'-O-tetramethylquercetin
Zhonglei WANG ; Liyan YANG ; Xiaohua ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1182-1187
This study was aimed to optimize optimum preparation process of 5,7,3',4'-O-tetramethylquercetin (TMQ). With 5,7,3',4'-O-tetramethylrutin as raw material, TMQ was prepared by acidic aqueous solution method. The preparation process of TMQ was optimized by single factor test and orthogonal test. The results showed that the optimum technology conditions were ethanol concentration at 0, temperature at 80℃, hydrochloric acid concentration at 0.50%, and the hydrolysis time for 1.0 h. It was concluded that the optimum preparation process had the charac-teristics of high yield, simple process, stable and feasible.
8.Comparative study on EEG and neuroimaging of patients with cerebral cysti-cercosis before and after treatment
Zhonglei WANG ; Tingxia FU ; Yingxin HU ; Guiling LI ; Dehua MAO ; Wenbin TAN
Chinese Journal of Schistosomiasis Control 2014;(3):311-313,319
Objective To study the changes of cerebral function and pathological morphology before and after the antiparasit-ic treatment with albendazole and praziquantel in patients with cerebral cysticercosis. Methods The data of EEG and neuroimag-ing of 412 patients with cerebral cysticercosis were retrospectively analyzed. Results Before the treatment,the mild abnormali-ty,moderate abnormality,and severe abnormality were observed in 40.53%,45.63% and 13.84% of the patients respectively, which mainly showed the diffuse or focal irregular slow waves,or epileptiform discharges found in the abnormal brain waves. CT/MRI manifestation could be divided into six types,including single sacculus type(23.59%),multiple sacculus type(44.42%), encephalitis type(13.59%),coexistence of macrocyst and sacculus type(4.85%),calcification type(2.18%),and mixed type (11.41%). After 3 courses of the treatment,the normal and improved EEGs were observed in 79.85%and 20.15%,respectively. CT/MRI showed the foci being all absorbed(77.18%),being most absorbed(20.63%),and being no changes(20.18%)which were calcified focus. When cerebral cysticercosis were in acute stage(the single and multiple sacculus type,encephalitis type, and macrocyst and sacculus coexistence type),the therapeutic effect was good;while in the mixed type,the therapeutic effect was relatively poor. If cysticercosis were in the calcification stage,the patients only needed the heteropathy. Conclusions In the pa-tients with cerebral cysticercosis,EEGs show the mild to severe abnormalities,and CT/MRI mainly shows the multiple sacculus type. After the treatment,the abnormal EEGs are gradually recovered and the low density foci can be all absorbed,but some calci-fied focus still exist in some patients.
9.Research Progress of Flavonoids with Antioxidant Activity Constituents from Chinese Materia Medica
Zhonglei WANG ; Xiaohua ZHANG ; Liyan YANG ; Xiangwei ZENG ; Pengshou LI ; Jiyang PAN ; Zhen YANG ; Jianna YUAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(3):551-554
Flavonoids, which is composed of a C6-C3-C6 backbone, is a kind of secondary metabolites of Chinese materia medica (CMM). It has received extensive attention from all over the world and obtained a rapid develop-ment due to its widespread distribution, structural complexity and active diversity. This article has summarized the progress of flavonoids both in antioxidant activity and in mechanism, so as to provide some references for the re-search and development of new medicine of flavonoids with antioxidant activity.
10.Standalone and Collaboration Application of New Technologies in Extraction of Chinese Medicine Polysaccharide Components
Zhonglei WANG ; Liyan YANG ; Xiangwei ZENG ; Xiaohua ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(6):1441-1446
With the modernization of traditional Chinese medicine (TCM), classic extraction methods such as sol-vent extraction, steam distillation, squeezing method, sublimation method have become much more difficult in sat-isfying needs of the development of the society as a result of low extraction rate of effective components, low per-centage impurity clearance. Therefore, in recent years the new extraction techniques with the advantages of green environmental protection , conditional stability , high extraction rate , strong biological activity , have been widely used in TCM extraction. These new extraction techniques have already shown each of its strength. The application progress on new extraction technology of Chinese medicine polysaccharide, both in standalone application and in collaboration application, were discussed in this article, so as to make new technologies be applied more reason-ably and effectively in Chinese medicine polysaccharide components.


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