Objective To analyze the nonlinear relationship between hypothermic machine perfusion (HMP) parameters and delayed graft function (DGF) and optimize the construction of a predictive model for DGF. Methods The data of 923 recipients who underwent kidney transplantation from deceased donors were retrospectively analyzed. According to the occurrence of DGF, the recipients were divided into DGF group (n=823) and non-DGF group (n=100). Donor data, HMP parameters and recipient data were analyzed for both groups. The nonlinear relationship between HMP parameters and the occurrence of DGF was explored based on restricted cubic splines (RCS). Over-sampling, under-sampling and balanced sampling were used to address the imbalance in the proportion of DGF to construct logistic regression predictive models. The area under the curve (AUC) of each model was compared in the validation set, and a nomogram model was constructed. Results Donor BMI, cold ischemia time of the donor kidney, and HMP parameters (initial and final pressures, resistance, and perfusion time) were significantly different between the DGF and non-DGF groups (all P<0.05). The RCS analysis revealed a threshold-like nonlinear relationship between HMP parameters and the risk of DGF. Among the models constructed using different sampling methods, the balanced sampling model had the highest AUC. Using this model, a nomogram was constructed to stratify recipients based on risk scores. Recipients in the high-risk group had higher serum creatinine levels at 1, 6, and 12 months after kidney transplantation compared to those in the low-risk group (all P<0.05). Conclusions There is a nonlinear relationship between HMP parameters and the risk of DGF, and the threshold is helpful for organ quality assessment and monitoring of graft function after transplantation. The predictive model for DGF constructed on the base of balanced sampling algorithms helps perioperative decision-making and postoperative graft function monitoring of kidney transplantation.

In recent years, a growing number of studies have shown that non-coding RNA plays an important role in the pathogenesis and development of epilepsy, and is involved in neuroinflammation, cell apoptosis, synaptic remodeling and cell differentiation in epilepsy. This review focused on the recent research progress in the relationship between microRNA, long-nocoding RNA and circular RNA with epilepsy.

Objective To explore the protective role of PR-39 on the ischemia reperfusion injury renal tissue of rats.Methods 60 male SD rats were enrolled and randomly divided into experimental KP group and negative control KE group.The rats orthotopic renal transplantation model were established.The recombinant adenoviral vectors containing promoter KSP and PR-39 gene (KP) or none (KE) were injected via renal artery.HE staining,immunohistochemistry and TUNEL staining were performed on the rats' renal tissues at 2 days after IRI modeling,and the renal tubular interstitial injury score and peritublar capillary rarefaction was also evaluated.The renal function related factors of rats was monitored before and after IRI,and the blood urea nitrogen and creatinine level was also compared between KP and KE groups.Results Blood urea and creatinine concentration was increased,part of glomerular and tubular injury in renal tissue were also visible in rats underwent renal IRI,which proved the rat orthotopic renal transplantation model were successfully estabilshed.PR-39 gene specifically expressed in renal interstitial tissue,but not in glomerular.The degree of kidney tublar injury and the number of apoptotic cells were lower in KP groups by comparing with negative control KE groups.Renal tubular injury was significantly decreased in group KP than in group KE.Peritubular capillary rarefaction index is smaller in KP than in KE group.The blood urea nitrogen and creatinine level in KP group were significantly lower than those in KE group,and the renal function were also recovered more quickly.Conclusion The specific expressed PR-39 in renal interstitial and tubule,can inhibit the apoptosis of renal tubular epithelial cells and promote the angiogenesis of peritubular capillary,so as to exert its protective role on IRI renal tissue.

Objective The magnetic beads direct adsorption method was used to extract the cell-free DNA (cfDNA) from three kinds of human humoral biological samples, including urine, saliva and blood, as to provide a reference for forensic cfDNA research and forensic inspection. Methods The cfDNA was isolated from humoral samples by centrifuging, and the cfDNA was extracted with the method of magnetic beads direct adsorption. Then the samples were sequentially amplified with Identifiler-Plus amplication kit, and the STR genotyping was detected by ABI 3500 Analyzer. Results The cfDNA was detected from all the three kinds of samples. The detection rate of cfDNA from the blood samples was 100%, the saliva was 90%, and the urine was 70%. Conclusion The results suggest that human humoral biological samples contain cfDNA. What's more, the magnetic beads direct adsorption method can be used to extract cfDNA efficiently and conveniently.

Objective To evaluate the effect of conversion from mycophenolic acid (MPA) to mizoribine (MZR) in renal transplant recipients with gastrointestinal tract (GI) symptoms.Methods A total of 355 renal transplant recipients with GI symptoms caused by MPA administration were enrolled from April 2015 to March 2017 in 25 different renal transplant centers in China.The symptomatic improvement of GI before (baseline) and after conversion to MZR (1,2,4 weeks) was assessed by each item of GI symptoms indication.In addition,the efficacy and safety of the conversion therapy during 12 months were determined.Results Patients showed improvement in GI symptoms including diarrhea,abdominal pain,abdominal distention and stomachache after conversion to MZR 1,2,4 weeks (P＜0.05).In patients with different severity of diarrhea,conversion to MZR therapy significantly improved diarrhea (P＜0.05).During 12 months,no patient experienced clinical immune rejection.We did not observe any infections,leucopenia and other serious side effects.Conclusion MZR could markedly improve GI symptoms caused by MPA administration in renal transplant recipients.

Objective To investigate the effects of IL-10/TGF-β-modified macrophages on renal ischemia reperfusion injury (IRI).Methods Bone marrow-derived macrophages were modified ex vivo by IL-10/TGF-β to acquire M2c (a subset of activated macrophages).M2c were transferred into treated C57BL/6 mice by a single tail-vein injection at 6 h after renal IRI.Mice were killed on the day 3 after renal IRI.Blood samples were collected to check renal function.Kidneys were harvested to determine tubular necrosis and apoptosis by H&E staining and TUNEL assay.Immunofluorescence was performed to analyze the proliferating tubular cell nuclear antigen.Meanwhile,proinflammatory cytokines and regulatory T cells in renal tissues were analyzed with real-time PCR and flow cytometry.Results In comparison with M1,M2c expressed lower levels of MHC Ⅱ (P＜0.01),CD86 (P＜ 0.01),TNFα (P＜0.01) and IL-1β (P＜0.01) and higher level of IL-10 (P＜0.01).M2c significantly attenuated renal functional decline (P＜0.01 or 0.05),structural injury (P＜0.05),apoptosis of tubular cells (P＜0.01) and inflammation factors infiltration (P＜0.01 or 0.05).What's more,the cells could promote tubular cells proliferation (P＜0.05) and regulatory T cells expression (P＜0.01).Conclusion Our results demonstrated that M2c macrophages effectively protect against renal IRI and may become a therapeutic strategy for renal IRI.

Fatty acid synthase (FAS) catalyses the reaction between acetyl-CoA and malonyl-CoA to produce fatty acids. It is one of the most important enzyme in lipid biosynthesis. FAS of the oleaginous yeast Rhodosporidium toruloides has two acyl carrier protein (ACP) domains and a distinct subunit composition compared with FASs of other species. As ACP is a protein cofactor crucial for fatty acid chain elongation, more ACPs in the FAS may facilitate the reaction. To study the biochemical and structural properties of this novel FAS from R. toruloides, plasmids were constructed and transformed into Escherichia coli BL21 (DE3). The strain ZWE06 harboring plasmids pET22b-FAS1 and pET24b-FAS2 could co-overexpress the two subunits. The recombinant FAS was purified by sequentially using ammonium sulphate precipitation, sucrose density gradient centrifugation and anion exchange chromatography. The specific activity of the recombinant FAS was 548 mU/mg. The purified complex would be used to study enzyme kinetics and protein structure of FAS, and heterogeneous expression and purification will facilitate revealing the mechanism of this novel FAS with double ACPs.
Acyl Carrier Protein ; Basidiomycota ; enzymology ; Chromatography ; Escherichia coli ; metabolism ; Fatty Acid Synthases ; biosynthesis ; genetics ; Fatty Acids ; biosynthesis ; Plasmids ; Recombinant Proteins ; biosynthesis ; genetics

Sclareol is a member of labdane type diterpenes mostly used as fragrance ingredient. To enable microbial production of sclareol, synthetic pathways were constructed by incorporating labdenediol diphosphate synthase (LPPS) and terpene synthase (TPS) of the plant Salvia sclarea into Saccharomyces cerevisiae. It was found that sclareol production could be benefited by overexpression of key enzyme for precursor biosynthesis, construction of fusion protein for substrate channeling, and removal of signal peptides from LPPS and TPS. Under optimal shake flask culture conditions, strain S6 produced 8.96 mg/L sclareol. These results provided useful information for development of heterologous hosts for production of terpenoids.
Alkyl and Aryl Transferases ; biosynthesis ; genetics ; Diterpenes ; metabolism ; Metabolic Engineering ; methods ; Metabolic Networks and Pathways ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Saccharomyces cerevisiae ; genetics ; metabolism ; Salvia ; chemistry ; enzymology ; genetics

To observe the effects of resveratrol on proliferation of human hepatocellular carcinoma cell line SMMC-7721 cells and expression of matrix metalloproteinase-9 (MMP-9) in vitro.

OBJECTIVE: To observe the efficacy of modified acupotome combined with blocking therapy in patients with carpal tunnel syndrome (CTS). METHODS: Fifty-five patients with CTS were divided into three groups, which were modified acupotome group including 26 CTS patients with 28 lesions treated by modified acupotome combined with blocking therapy, traditional acupotome group including 14 CTS patients with 16 lesions treated by traditional acupotome combined with blocking therapy, and blocking therapy group including 15 CTS patients with 15 lesions only treated by local blocking. The treatment outcome and one-year recurrence rate were observed. RESULTS: The response rate and one-year recurrence rate after operation in the modified acupotome group were 85.7% (24/28) and 20.8% (5/24) respectively, which had no significant differences as compared with 81.3% (13/16) and 38.5% (5/13) in the traditional acupotome group. The response rate and one-year recurrence rate after operation in the above two groups were both improved significantly as compared with those in the blocking therapy group which were 46.7% (7/15) and 85.7% (6/7) respectively. There were no acupotome-related adverse effects and injuries observed in the modified acupotome group. CONCLUSION: The modified acupotome is a considerable treatment method for CTS with respect to its simple manipulation and high effectiveness.