1.Surface labeling of bone marrow mesenchymal stem cells by biotin-streptavidin
Lin YANG ; Fuli LUO ; Yun LI ; Jun WEN ; Yang XU
Chinese Journal of Tissue Engineering Research 2016;20(10):1382-1388
BACKGROUND:Currently, there is a lack of efficient, non-invasive way to transplant stem cels to the target organ or tissue. Exploring a way to guide targeting transplantation of stem cels and to improve the efficiency of stem cel homing is now one of focuses in the field of stem cels research.
OBJECTIVE: To establish a simple and feasible method to chemicaly modify the cel surface using biotin-streptavidin reaction system, and to evaluate the efficiency of this method to label bone marrow mesenchymal stem cels (BMSCs) and its effects on cel biological functions.
METHODS: Passage 3 BMSCs were obtained by whole bone marrow culture method and verified by flow cytometry. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were used to equip the adhesion molecule ligand, sialyated LewisX (SLeX), to the BMSCs surface. The labeling rate of BMSCs was assessed using fluorescence microscope, the vitality of BMSCs was evaluated by trypan blue staining, and the proliferation of BMSCs was evaluated by cel counting kit-8 assay. Adipogenic and osteogenic inductions were used to evaluate the effect of the method on the multi-differentiation function of BMSCs.
RESULTS AND CONCLUSION: After culture for 2 weeks, passage 3 BMSCs were obtained and confirmed by expressing CD90, CD29 and lack of CD34, CD45. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were successfuly used to equip sialyated LewisX (SLeX) to the BMSCs surface and had minor effects on the vitality, proliferation, and differentiation of BMSCs. This method was simple for surface modification and had a high modification rate of 88%. The homing of BMSCs modified by this method to the target organ or tissue could be greatly enhanced. Therefore, this method potentialy could have extensive and important applications.
2.Meta-analysis of the efficacy of two-compartment peritoneal dialysis fluid with low glucose degradation products in peritoneal dialysis patients
Fuli LUO ; Dehan CAI ; Lin YANG ; Yun LI
Chinese Journal of Nephrology 2014;30(9):671-678
Objectives To study the efficacy of the two-compartment peritoneal dialysis fluid with low glucose degradation products in peritoneal dialysis (PD) patients.Methods Pubmed,EBMASE,Cochrane Library,Wanfang,VIP,CNKI,CBM and other databases were searched,at the same time the information form relevant literatures until December 2013 were searched by hand.To be eligible,studies had to be randomized controlled trials that allocated PD patients to two-compartment peritoneal dialysis fluid with low glucose degradation products (low-GPDs group) or to traditional dialysis fluid (control group).The qualities of included articles were assessed and then a meta-analysis was conducted by using RevMan 5.2 software.Results A total of 12 documents,11 studies met the inclusion criteria,and 1 059 cases were included.Meta-analysis results were as follows:(1)the low-GPDs group had higher level of CA125 in peritoneal dialysis effluent,higher residual renal function compared with that in the control group and the weighted mean difference were 19.61 (95%CI 12.04-27.18,P < 0.01) and 0.78 (0.14-1.43,P=0.02),respectively; (2)There was no statistically significant difference between control and low-GPDs group in the ultrafiltration,peritonitis and plasma bicarbonate (all P > 0.05); (3)Four studies showed no difference in peritoneal dialysis technique survival between the two group (P > 0.05).Conclusions The two-compartment peritoneal dialysis fluid with low glucose degradation products is effective and safe,has no negative effects on the frequency of peritonitis,patient' s peritoneal member transport function and plasma bicarbonate,but it causes less mesothelial damage and has higher residual renal function in patients than conventional ones,and does not affect the technique survival time.
3.Bilateral stereotactic radiofrequency amygdalohippocampectomy for the treatment of bilateral medial temporal lobe epilepsy
Quanjun ZHAO ; Huimin LUO ; Zengmin TIANA ; Zhaohui WU ; Fuli WANG ; Feng YIN ; Hulin ZHAO ; Haiying WANG
The Journal of Practical Medicine 2014;(6):853-856
Objective To retrospectively analyze the effect and safety of bilateral stereotactic radiofrequency amygdalohippocampectomy (SAHE) for treatment of bilateral medial temporal lobe epilepsy (BMTLE). Methods Twelve BMTLE patients were treated with bilateral SAHE under limited coagulations. Clinical parameters were evaluated with the programs of Engel′s classification, Liverpool Seizure Severity Scale (LSSS) 2.0, Wechsler Adult Intelligence Scale-Revised (WAIS-R) and Wechsler Memory Scale-Revised (WMS-R), respectively. Results Five patients (42%) were assessed as EngelⅠwith 12 ~ 62-month follow-up. Seizure severity scores were declined sharply compared with the baseline of the patients with out seizure free. Function of memory and intelligence was transiently declined without statistical significance immediately after operation (P >0.05), but was significantly increasedat 6 months after operation (P < 0.05). Conclusion Bilateral SAHE could terminate seizures or reduce seizure severity in patients with BMTLE. Under the circumstance of limited coagulations, neuropsychological function was improved along with seizure control.
4.Preparation and characterization of sustained-release levofloxacin bone tissue-engineered three-dimensional silk fibroin/chitosan/nano-hydroxyapatite scaffold
Peng YE ; Fuli LUO ; Anping LIU ; Haizhen DUAN ; Quan HU ; Wenjin HUANG ; Yun CHENG ; Anyong YU
Chinese Journal of Tissue Engineering Research 2019;23(14):2147-2155
BACKGROUND: Preliminary study has prepared the three-dimensional silk fibroin/chitosan/nano-hydroxyapatite scaffold successfully.OBJECTIVE: To explore the mechanical properties, physical characteristics, chemical composition and antibiotic sustained-release ability of three-dimensional silk fibroin/chitosan/nano-hydroxyapatite scaffold loaded with levofloxacin. METHODS: Levofloxacin/chitosan (3:1) microspheres were constructed by emulsion settlement filter method. 5, 7.5 and 10 g of microspheres were added into 2% of silk fibroin/chitosan/nano-hydroxyapatite mixed solution through freeze drying and chemical cross-linking to obtain the scaffolds loaded with antibiotics. The scaffolds loaded with antibiotics underwent scanning electron microscope observation, and chemical composition analysis. The sustained release, mechanical properties, porosity, water absorption expansion rate and hot water soluble loss rate were detected. RESULTS AND CONCLUSION: (1) Scanning electron microscope observed that there were drug microspheres at the inner wall of the scaffold, and the voidage was decreased with mass of microspheres increasing. (2) Energy spectrum analysis showed that the three kinds of scaffolds were rich in calcium and phosphonium ions. (3) The three kinds of scaffolds showed the same releasing trend, which presented with sudden-release effect at the former 3 days (release> 50%) , and then tended to be stable. The release rate was the slowest in the scaffold loaded with 10 g of microscopes, and the rapidest in the scaffold loaded with 5 g of microscopes. (4) With the mass of microspheres increasing, there was an increase in the compressive and tension abilities and hot water soluble loss rate, and a decrease in the porosity, mean pore size and water absorption expansion rate. (5) These results indicate that the three-dimensional tissue-engineered scaffold loaded with levofloxacin is constructed successfully by freeze drying and chemical cross-linking method, which holds good sustained-release effect and compressive ability, water absorption expansion rate and hot water soluble loss rate.
5.Cervical heart transplantation in mice model
CAO Jianbin ; GONG Yongquan ; WEI Chengxin ; LI Fuli ; LUO Peng
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2017;24(11):886-889
Objective To investigate how to establish stable mice cervical heart transplantation model. Methods Totally, 40 male C57 mice with the age of 6-8 weeks and weight of 19-24 g were randomly divided into recipients and donors (n=20 in each group). Mice cervical heart transplantation model was established by connecting the ascending aorta of donors to the right cervical common artery of recipients through end to side anastmosis and the pulmonary artery of donors to the right external jugular vein of recipients through end to end anastmosis. Results More than 95% recipients survived after surgery. Cold ischemia time was 15±5 min, warm ischemia time 23±6 min, and the whole operation took about 55±15 min. The recipients survived more than 30 d with functional heart grafts. Histologically, there was no difference between the heart graft one month after the transplantion and the normal heart. Conclusion Cervical heart transplantation of mice model is reliable and feasible, which is easy to monitor the survival condition of heart graft by visual examination and palpation, which will benefit the basic research in transplantation field.
6.Anatomical location as a prognostic factor in surgical treatment of gallbladder carcinoma
Zhencheng ZHU ; Kunlun LUO ; Bin WU ; Hong LIU ; Zheng FANG ; Yang BAI ; Fuli LI ; Weiwei LIU ; Liuqun SHAN
Chinese Journal of Hepatobiliary Surgery 2020;26(8):600-605
Objective:To explore the prognostic factors of patients with gallbladder cancer in different anatomical locations.Methods:A retrospective analysis of the clinical data of 140 patients with gallbladder cancer who were surgically treated in the 904 Hospital of the Joint Logistics Support Force from January 2015 to December 2015. Among them, 49 were males and 91 were females, with a median age of 64.0 years. According to the location of gallbladder cancer, they were divided into the neck of the gallbladder and the bottom of the gallbladder. The clinical data of the two groups were compared. The Kaplan-Meier method was used for univariate analysis, and Cox multivariate regression analysis was used to analyze the significant factors of univariate survival to determine the independent risk factors that affect the prognosis of patients with gallbladder cancer. Use Pearson correlation to analyze the correlation between clinicopathological characteristics.Results:Patients in the neck of the gallbladder group had higher levels of preoperative total bilirubin, preoperative albumin, carcinoembryonic antigen, carbohydrate antigen 19-9, extrahepatic bile duct resection (biliary-enteric anastomosis), radical resection, breakthrough of the liver/serous membrane invasion, lymph node metastasis, TNM staging, local recurrence in the operation area in the surgical area were statistically significant ( P<0.05). Univariate analysis suggested that preoperative jaundice, Glasgow prognostic score (GPS), C-reactive protein, carcinoembryonic antigen, carbohydrate antigen 19-9, accidental gallbladder cancer, surgical method, tumor pathological type, tumor differentiation degree, tumor growth location (neck than the bottom of the body) are related to the prognosis (all P<0.05). Cox regression analysis indicated that GPS ( RR=3.272, 95 CI: 1.987-5.388), surgical method ( RR=4.149, 95 CI: 2.561-6.723), tumor location ( RR=0.316, 95 CI: 0.209-0.478), distant metastasis ( RR=1.695, 95 CI: 1.036-2.775) and TNM staging ( RR=3.686, 95 CI: 2.222-6.115) are independent risk factors affecting the prognosis of gallbladder cancer. Correlation analysis suggests that neck tumors are related to later staging, liver bile duct invasion, lymph node metastasis, high inflammation levels, and low radical cure rates. Conclusions:Gallbladder cancer that occurs in the neck of the gallbladder is an independent factor influencing the poor prognosis of patients with surgical treatment. In addition, TNM staging, distant metastasis and Glasgow score are important predictors of survival in patients with gallbladder cancer.
7.Expression of GOLPH3 and NLRP3 in gallbladder carcinoma and their clinical significance
Zhencheng ZHU ; Kunlun LUO ; Hong LIU ; Zheng FANG ; Fuli LI ; Yang BAI ; Weiwei LIU ; Liuqun SHAN ; Haibin ZHAO
Chinese Journal of Hepatobiliary Surgery 2020;26(11):846-849
Objective:To investigate the expression and clinical significance of Golgi phosphoprotein 3 (GOLPH3) and NOD-like receptor protein 3 (NLRP3) in gallbladder carcinoma (GBC).Methods:Surgical specimens and clinical data were collected from 63 patients with GBC who underwent radical cholecystectomy in the 904th Hospital of Joint Logistic Support Force of PLA from January 2014 to January 2019. In the GBC group, there were 21 males and 42 females, with an average age of 62.5 years. For 30 patients with mild to moderate atypical hyperplasia of gallbladder during the same period were included in the precancerous lesion group, including 9 males and 21 females, with an average age of 62.4 years. Normal gallbladder specimens from 20 patients who underwent surgical resection due to liver trauma or giant hepatic hemangioma were collected and included in the normal group, including 7 males and 13 females, with an average age of 61.9 years. The expressions of GOLPH3, NLRP3, Ki-67 were detected by immunohistochemistry. Log-rank test and Cox regression were adopted to analyze the GOLPH3 and NLRP3 expression and survival prognosis of gallbladder cancer patients.Results:Expression of GOLPH3 and NLRP3 in the tumor group, precancerous lesion group and normal group was decreased separately. In GBC tissues, the level of GOLPH3 and NLRP3 was positively correlated with the Ki-67 expression ( r=0.972 and r=0.969, both P<0.05). Multivariate analysis showed that high level of GOLPH3 ( HR=4.891, 95% CI: 1.776-13.470) and NLRP3 ( HR=3.006, 95% CI: 1.273-7.099) was an independent risk factor for predicting the postoperative survival of patients with GBC (both P<0.05). Conclusion:GOLPH3 and NLRP3 are highly expressed in GBC tissues, and high expression of GOLPH3 and NLRP3 is an independent risk factor for postoperative survival in patients with GBC.
8.Mechanism of Shenshuai Xiezhuo Decoction in Intervention of Chronic Kidney Disease Combined with Atherosclerosis Based on Regulation of PI3K/Akt/mTOR Signaling Pathway by miRNA126
Gengxin LIU ; Gedi ZHANG ; Wenqi YUAN ; Fusheng LI ; Min GUO ; Fuli LUO ; Ziyou YAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):48-55
ObjectiveExploring the role of microRNA126 (miRNA126) in chronic kidney disease combined with atherosclerosis (CKD AS) by regulating the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway and the mechanism of Shenshuai Xiezhuo decoction in the intervention of CKD AS rats with 5/6 nephrectomy combined with high-fat feeding. MethodA total of 60 SD rats were randomly divided into sham operation group, model group, losartan group, and low, medium, and high dose groups of Shenshuai Xiezhuo decoction. The CKD AS rat model was established by 5/6 nephrectomy combined with high-fat feeding for 10 weeks. The low, medium, and high dose groups (6.0, 12.0, 24.0 g·kg-1·d-1) of Shenshuai Xiezhuo decoction and the losartan group (20 mg·kg-1·d-1) were gavaged, and the corresponding intervention was carried out for eight weeks. Then, the rats were killed, and samples were collected for corresponding detection. Fully automated biochemical analyzers were used to detect kidney function and blood lipids in rats: blood creatinine (SCr), blood urea nitrogen (BUN), total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) levels. Hematoxylin-eosin (HE) and Masson staining of aortic tissue and pathological observation under a light microscope were carried out, and autophagosomes and autophagy lysosomes were observed by transmission electron microscopy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to determine the mRNA levels of miRNA126, PI3K, Akt, and mTOR in rats, and Western blot was used to determine the protein expression levels of phosphorylated (p)-PI3K, PI3K, p-Akt, Akt, p -mTOR, mTOR, benzyl chloride 1 (Beclin-1), and microtubule-associated protein light chain 3Ⅱ/Ⅰ (LC3Ⅱ/LC3Ⅰ). ResultCompared with the sham operation group, the serum SCr, BUN, TC, TG, and LDL-C in the model group were significantly increased (P<0.01). Compared with the model group, the SCr, BUN, TC, TG, and LDL-C were decreased in the losartan group and low, medium, and high dose groups of Shenshuai Xiezhuo decoction (P<0.05). Compared with the sham operation group, thickening plaques, infiltration of mononuclear macrophages, a small number of foam cells, disordered arrangement of smooth muscle fibers in the tunica media, and increased collagen fibers were observed in the model group, and the lesions in the losartan group and Shenshuai Xiezhuo decoction groups were alleviated compared with those in the model group. Compared with the model group, the number of autophagosomes and autophagy lysosomes increased in the medium and high dose groups of Shenshuai Xiezhuo decoction. Compared with the sham operation group, the expression of miRNA126 in the aortic tissue of the model group was significantly decreased (P<0.01), and the mRNA expressions of PI3K, Akt, and mTOR were significantly increased (P<0.01). Compared with the model group, the expression of miRNA126 in the aortic tissue of rats in high, medium, and low dose groups of Shenshuai Xiezhuo decoction and losartan group was significantly increased (P<0.01), while the mRNA expressions of PI3K, Akt, and mTOR were significantly decreased (P<0.01). Compared with the sham operation group, the protein expressions of p-PI3K, PI3K, p-Akt, Akt, p-mTOR, and mTOR in the model group were significantly increased (P<0.01), while the protein levels of Beclin-1, LC3Ⅰ, and LC3Ⅱ were significantly decreased (P<0.01). Compared with the model group, the protein expressions of p-PI3K, PI3K, p-Akt, Akt, p-mTOR, and mTOR in the losartan group and low, medium, and high dose groups of Shenshuai Xiezhuo decoction were decreased (P<0.05), while the protein levels of Beclin-1 and LC3Ⅱ/LC3Ⅰ were increased (P<0.05). ConclusionThe expression of miRNA126 is decreased in the aortic tissue of CKD AS rats, and the PI3K/Akt/mTOR pathway is activated to inhibit autophagy flux. Shenshuai Xiezhuo decoction regulates the PI3K/Akt/mTOR signaling pathway through miRNA126, restores the autophagy of aortic endothelial cells, protects the damage of CKD vessels, reduces the formation of As plaques, and slows the development of cardiovascular complications.