1.Recombinant mycobacterium tuberculosis heat shock protein 10 in human osteoclast differentiation
Yuanyu ZHANG ; Yongrong GUO ; Xia LIU ; Kun LI
Chinese Journal of Tissue Engineering Research 2014;(38):6116-6122
BACKGROUND:The mycobacterium tuberculosis heat shock protein 10 exerts effects on the osteoclasts by in vitro mouse cranium experiment,
OBJECTIVE:To investigate the effect and mechanism of recombinant mycobacterium tuberculosis heat shock protein 10 (CPN10) on the differentiation of osteoclasts in the in vitro culture system that induces osteoclast differentiation.
METHODHuman macrophage colony-stimulating factor-dependent adhesive blood mononuclear cells were divided into four groupreceptor activator for nuclear factor-κB ligand (RANKL)+CPN10 (1 mg/L), RANKL, CPN10 (1 mg/L), and negative control (complete culture medium). Monocytes were resuspended in a-MEM medium containing macrophage colony-stimulating factor, and were cultured in each group for 7, 14, 21 days. The morphology, quantity and bone resorption area of osteoclasts were examined by tartrate-resistant acid phosphatase (TRAP) staining. The expressions of NFATc1 and c-Fos gene and protein were also detected.
RESULTS AND CONCLUSION:In negative control group, no TRAP-positive multinucleated osteoclasts generated, while in the other groups, TRAP-positive multinucleated osteoclasts differentiated and formed the lacunae in the smal bone grinding. The number of osteoclasts formation and resorption in CPN10 group were significantly lower than that in RANKL+CPN10 group. The expression of NFATc1 and c-Fos in the negative control group C was significantly lower than that of RANKL+CPN10 group and CPN10 group. However, CPN10 expressed NFATc1 and c-Fos protein, which was significantly lower than RANKL+CPN10 group. CPN10 is involved in the formation of osteoclasts, and the mechanism is related with the upregulation of NFATc1, c-Fos expression.
2.Effect of recombinant Mycobacterium tuberculosis heat shock protein 10 on proliferation of human osteoblasts and regulation of bone metabolism
Yuanyu ZHANG ; Xia LIU ; Kun LI ; Yongrong GUO ; Jingping BAI
Chinese Journal of Tissue Engineering Research 2014;(11):1665-1671
BACKGROUND:Mycobacterium tuberculosis heat shock protein 10 (r-Mt cpn10) is one of the main factors that cause bone tuberculosis dissolution and absorption as wel as inhibits the proliferation of osteoblasts. Receptor activator of nuclear factor kappa B ligand and osteoprotegerin are the important factors influencing bone metabolism.
OBJECTIVE:To observe the effect of r-Mt cpn10 on human osteoblast proliferation, alkaline phosphatase secretion, expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA. METHODS:Human bone marrow stromal cel s were induced to differentiate into osteoblasts, and osteoblasts at passage 3 were cultured with various concentrations of r-Mt cpn10 (0.1, 1, 10 mg/L). Osteoblasts cultured without r-Mt CPN10 were assigned as controls.
RESULTS AND CONCLUSION:MTT assay results showed that, compared with control group, r-Mt cpn10 at different concentrations inhibited osteoblast proliferation and alkaline phosphatase secretion (P<0.05). RT-PCR analysis showed that, r-Mt cpn10 at different concentrations increased receptor activator of nuclear factor-kappa B ligand mRNA expression (P<0.01), and inhibited osteoprotegerin mRNA expression in a concentration-dependent manner (P<0.01). 10 mg/L r-Mt cpn10 exhibited the strongest effect (P<0.01). The r-Mt cpn10 can inhibit osteoblast proliferation and alkaline phosphatase activity, and it may influence bone metabolism by regulating the expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA.
3.Distribution and antibiotic resistance of pathogens isolated from inpatients with urinary tract infections
Yuanyu GUO ; Heyue LEI ; Qinqin CHEN ; Guoxiang ZHANG
Chinese Journal of Clinical Infectious Diseases 2012;(6):350-354
Objective To investigate the distribution and antibiotic resistance of pathogens isolated from inpatients with urinary tract infections.Methods A retrospective analysis was carried out on 1033 strains of pathogens isolated from urine culture in patients with urinary tract infections in Zhejiang Xiaoshan Hospital during January 2009 and December 2011.Urine specimens were cultured with Uricult,and K-B method was used for drug susceptibility test,WHONET 5.6 software was used to analyse drug susceptibility test.Results Among 1033 strains of pathogens,681 (65.9%) were gram-negative bacteria,197 (19.1%) were gram-positive bacteria,and 155 (15.0%) were fungi.The three most prevalent bacteria were Escherichia coli (402 strains,38.9%),Klebsiella pneumoniae (74 strains,7.2%) and Candida albicans (64 strains,6.2%).60.7% (244/402) of Escherichia coli and 45.9% (34/74) of Klebsiella pneumoniae were extended-spectrum β-lactamases (ESBLs) positive.Escherichia coli and Klebsiella pneumoniae were susceptible to imipenem,meropenem,cefoperazone/sulbactam,piperacillin/tazobactam and amikacin.Enterococcus and staphylococcus were susceptible to vancomycin,linezolid and furadantin.Candida was sensitive to flucytosine,voriconazole and amphotericin B.Conclusion Gram-negative bacteria mainly E.coli is the predominant pathogen to urinary tract infections in this group of patients.Regular analysis and monitoring of pathogen species and drug resistance is important for rational use of antibiotics.
5.Genotypes of β-lactamases produced by Klebsiella pneumoniae
Yuanyu GUO ; Zeqing WEI ; Peiqiong ZHU ; Xiaoxing DU ; Qing YANG ; Ping SHEN ; Yunsong YU
Chinese Journal of Clinical Infectious Diseases 2010;3(3):138-141
Objective To investigate the genotypes of β-lactamases produced by Klebsiella pneumoniae.Methods Plasmid conjugation,PCR amplification,gene cloning and DNA sequencing,isoelectric focusing electrophoresis and extended-spectrum β-lactamase(ESBLs)confirmatory test were carried out for analyzing the encoding gene of β-lactamases in clinical strains of Klebsiella pneumoniae collected from hospital wards.Results Totally 75 clinical strains of Klebsiella pneumoniae were collected,in which 48 strains were confirmed to produce genotype of β-laetamases(64.0%),including 39 ESBLs-producing Btraim(52.0%).Among 48 strains,17 isolates(35.4%)carried 2 types of ESBLs genes,7(14.6%)carried 3 types of ESBL8 genes,and 5(10.4%)carried 4 types of ESBLs genes.CTX-M was the most comon type(30/48,62.5%),followed by TEM(26/48,54.2%)and SHV(25/48,52.1%).Among 9 isolates with DHA-1 AmpC β-laetamase,8 produced AmpC β-lactamases and ESBLs.Class A carbapenemase KPC-2 was produced in 3 isolates.False negative rate of ESBLs confirmatory test was 23.1%(9/39).Condusion Genotypes of β-lactamases produced by Klebsiella pneumoniae are complicated,which results in multi-drug resistance in clinic.
6.Application of en-bloc mesogastric excision in the treatment of advanced gastric cancer.
Fujian JI ; Xuedong FANG ; Junnan JIANG ; Yuanyu WU ; Ye FENG ; Huiling GUO
Chinese Journal of Gastrointestinal Surgery 2016;19(10):1097-1100
OBJECTIVETo evaluate the value of en-bloc mesogastric excision (EME) in the treatment of advanced gastric cancer.
METHODSA retrospective analysis on clinical data of 98 gastric cancer patients who underwent total gastrectomy in China-Japan Union Hospital of Jilin University from January 2013 to December 2015 was carried out, including EME group of 48 cases (according to the mesangial space) and D2 radical group of 50 cases(D2 lymphadenectomy according to the vascular markers). Operations were performed by the same single surgeon team. Surgical indexes and recent efficacy indexes were compared between two groups.
RESULTSGeneral informations pertaining to two groups were comparable (P>0.05). All the operations were performed successfully. Compared with D2 radical group, EME group had a shorter operative time [(155.3±13.6) vs. (171.2±14.9) minutes, P=0.012] and less intraoperative blood loss [(95.1±19.5) vs.(122.6±28.0) milliliters, P=0.011]. There were no significant differences in the number of harvested lymph node (30.8±3.9 vs. 31.5±4.7, P=0.675), time to postoperative bowel function return [(3.2±1.2) vs.(3.9±1.4) days, P=0.179], postoperative hospital stay [(10.9±2.7) vs.(11.3±3.2) days, P=0.788], and the incidence of postoperative complication [8.3% vs. 10.0%, P=0.775]. During the follow-up of 1 year, all the patients had no long-term complications, no tumor recurrence or death.
CONCLUSIONSFor advanced gastric cancer, EME result in the same clinical efficacy compared with standard D2 resection. At the same time, EME can shorten the operative time and reduce the intraoperative blood loss, which is a new technology and worthy promoting.
Aged ; Blood Loss, Surgical ; Defecation ; Female ; Gastrectomy ; methods ; Humans ; Laparoscopy ; Length of Stay ; Lymph Node Excision ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Operative Time ; Postoperative Complications ; Postoperative Period ; Retrospective Studies ; Stomach Neoplasms ; surgery ; Treatment Outcome
7.Inhibition of islet allograft rejection by Qa-1/PD-L1 artificial liposome
Meng GUO ; Yuanyu ZHAO ; Hao YIN ; Jiayong DONG ; Junsong JI ; Qi LU ; Hang YUAN ; Fei TENG ; Wenyuan GUO ; Guoshan DING
Chinese Journal of Organ Transplantation 2019;40(2):72-77
Objective To explore the effects of Qa-1 and PD-L1 loaded artificial liposomal treatment in allograft rejection and its outcomes .Methods The extracellular domains of Qa-1 and PD-L1 were loaded on liposome surface by streptavidin-biotin system . Mixed lymphocyte reaction (MLR) was performed for measuring Qa-1/PD-L1 liposome biological function .Then liposome was co-transplanted with allo-islets via portal vein .The levels of blood glucose and C-peptide were detected daily after transplantation .Also hepatic lymphocytes after transplantation were isolated for determining the proportion of activated cells and signaling pathway changes .Results Artificial liposome could be easily loaded with biotinylated peptide and its diameter was between 50 to 500 nm . Qa-1/PD-L1 liposome could significantly suppress lymphocyte proliferation , activation and secretion of IFN-γ in MLR by an activation of SHP1/2 and an inhibition of Syk pathway .Qa-1/ PD-L1 liposomes could suppress the activation of hepatic lymphocytes in vivo by activating SHP1/2 ,protecting islet allografts and maintaining a normal level of blood glucose in recipients .Conclusions Qa-1/PD-L1 loaded liposome can effectively suppress allograft rejection and improve the outcomes of islet transplantation .
8.Effect of adoptive reinfusion of Treg on immune rejection of islet allografts in mice
Junhui LI ; Yuanyu ZHAO ; Meng GUO ; Junsong JI ; Hang YUAN ; Hui WANG ; Qi LU ; Zhiren FU ; Guoshan DING ; Hao YIN
Organ Transplantation 2019;10(6):690-
Objective To investigate the effects of adoptive reinfusion of regulatory T cell (Treg) on the recovery of islet function and graft survival time after islet allograft transplantation. Methods The diabetic model was established using C57BL/6 mice as recipients, and Balb/c mice were chosen as donors for islet allografts transplantation beneath the renal capsule. The recipient mice were divided into 3 groups and 3 mice in each group according to different processing Methods: Treg experiment group (Treg group, 1×106 Treg cells were injected via tail vein at 1 d before operation), positive control group [sirolimus (SRL) group, SRL at a dose of 300 μg/(kg·d) was intragastrically given every day from 1 d before operation] and blank control group (control group, an equivalent volume of normal saline was intragastrically given every day from 1 d before operation). Enzyme-linked immune absorbent assay (ELISA) was used to detect the changes of blood glucose and C-peptide in mice within 14 days after transplantation.
9.Research advances in total pancreatectomy with autologous islet cell transplantation
Jingjing LI ; Yuanyu ZHAO ; Hong FU ; Fei TENG ; Jiaxi MAO ; Wenyuan GUO
Journal of Clinical Hepatology 2023;39(10):2506-2512
Since the 1970s, patients with chronic pancreatitis (CP) have benefited from total pancreatectomy with autologous islet cell transplantation (TPAIT). With the continuous development of surgical techniques and perioperative management over the past few decades, there have been improvements in islet cell function, insulin independence rate, and the survival rate of patients. This article summarizes the preoperative indications for TPAIT, the development of surgical operations, postoperative management and monitoring, and prognosis, so as to help clinicians learn more about TPAIT.
10.Opportunities and challenges of marginal donor liver
Xinyi LU ; Fei TENG ; Hong FU ; Yuanyu ZHAO ; Liye ZHU ; Jiayong DONG ; Jiaxi MAO ; Wenyuan GUO
Organ Transplantation 2024;15(3):463-468
With persistent breakthrough and maturity of surgical procedures and postoperative immunosuppressive therapy, the survival rate of liver transplant recipients and grafts has been significantly increased. The shortage of donor liver has become the main obstacle for clinical development of liver transplantation. How to expand the source of donor liver has become an urgent issue. Groundbreaking progresses have been made in the use of common marginal donor livers in clinical liver transplantation, such as elderly donor liver, steatosis donor liver, viral hepatitis donor liver and liver from donation after cardiac death. Nevertheless, multiple restrictions still exist regarding the use of marginal donor liver. Consequently, the definition of marginal donor liver and research progress in the application of common marginal donor livers were reviewed, and the opportunities and challenges of mariginal donoor liver were illustrated, aiming to provide reference for expanding the donor pool for clinical liver transplantation and bringing benefits to more patients with end-stage liver disease.