Humans ; Neoplasms ; genetics ; immunology ; therapy ; RNA Interference ; RNA, Small Interfering ; RNA-Induced Silencing Complex

Animals ; Antioxidants ; pharmacology ; Ascorbic Acid ; pharmacology ; Cumulative Trauma Disorders ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Muscle, Skeletal ; injuries ; pathology ; Oxidation-Reduction ; Oxidative Stress ; drug effects ; Rats

Objective: To express glycosyl-phosphatidylinositol (GPI) modified Met- RANTES fusion protein on Chinese hamster ovary (CHO) cells and to develop a novel immunosuppressant GPI anchored form of Met-RANTES. Methods: The eukaryotic expression vector PEF/GPI-Met-RANTES were constructed and transfected into CHO cells by electroporation. The transfectants were selected with methotrexate (MTX). Expression of the recombinant protein was assessed by flow cytometric analysis, cell immunofluorescence staining and immunogold electron microscopy. Results: The chimeric molecules of GPI anchored form of Met-RANTES including the whole reading frame were constructed, and the sequence was identical to the designed sequence. GPI anchored form of Met-RANTES was stably expressed on CHO- DHFR- cells. Conclusion: A large amount of GPI modified Met-RANTES fusion protein was expressed on CHO cells. GPI anchored form of Met-RANTES may be used as novel immunosuppressant for suppressing reaction in graft rejection.

Zinc-fingers and homeoboxes 2 (ZHX2) is a novel transcriptional repression factor participating in regulating the expression of alpha fetal protein (AFP). ZHX2 has been confirmed to be largely correlated with development and metastasis of hepatocellular carcinoma (HCC) and invasion of multiple myeloma.

[Objective] To study the role of scaffold internal three dimensional structure of on vascularization of ?-TCP biomaterials in vivo.[Method]Twenty four adult rabbits were selected for operation.Two different three-dimensional structure ?-TCP biomaterials(wafer ?-TCP,the pore diameter was from 400 ?m to 500 ?m,the pore inter-connection diameter was 120 ?m;granulation ?-TCP,the particle diameter was from 100 ?m to 200 ?m)were implanted separately into fascia lumbodorsalis of every rabbit.The specimens were harvested in 1,2,4,8 weeks after surgery for histology,scanning electronic microscope(SEM)and SPECT studies in order to observe the vascularization of two different structure ?-TCP biomaterials in vivo.[Result]The biocompatibility of two different ?-TCP biomaterials was favourable.Only a few of immature blood capillaries were detected in some peripheral pores of two different biomaterials in one week after surgery.In four weeks of implantation,the result of histology indicated that the wafer artificial bone had vascularized completely.The number and lumens of blood vessel had increased.The peripheral blood vessel had been mature,showing vascularization crest-time.In eight weeks after sugery,there was no more increase of the number of blood vessel,while the lumens of blood vessel had increased.The mature capillaries were observed by chance.To compare with the wafer artificial bone,the vas cularization rate of the granulation artificial bone biomaterials was slower,and the number of blood vessel was less.On the other hand,the smaller lumens diameter and the infant structure existed in most of blood capillaries.Many blood vessels were not mature in four weeks,the vascular occlusion in some pores was detected.[Conclusion]The pore interconnection pathway in scaffolds is a key factor for vascularization.In other words,the higher density of pore interconnection pathway can induct more complete vascularization in scaffolds,and the diameter can restrict the lumens of blood vessel diameter.

The “vertical tongue”method was used in speech training for 12 patients with functional speech disorder of consonant /L/.Af-ter treatment,the average vocal intilligibility of the 12 patients increased from 86.3% to 98.9%(P <0.05)./L/consonant average intelligi-bility increased from 42.9% to 85.2%(P <0.05).

Objective:To investigate the effect of genistein,a soybean-derived isoflavone,on thestimulating effect on bone resorption of IL-1?.Methods:The osteoclasts(OCs)were isolated with themethods of Yu Shifeng.The rat calvaria were cultured as an organ.The cells in the experiment weregrown in four respectively:Control(without genistein or IL-1?),10~(-6) mol/L genistein,10 ?g/L IL-1?and 10~(-6) mol/L genistein+10 ?g/L IL-1?.The area of bone resorption,the concentration of Ca~(2+) in thesupernatant liquid of OCs cultures and mice calvaria were tested.The contents of acid phosphatase(ACP)were also examined by biochemistry method.The index of bone resorption was counted as the ra-tio of the experiment average and control ones,which indicated the increase in bone resorption when itwas above 1.0.Results:The area of bone resorption of 10~(-6) mol/L genistein+10 ?g/L IL-1? increasedcompared with that of 10~(-6) mol/L genistein,while the concentration of Ca~(2+) in the supernatant liquid ofOCs cultures decreased significantly.The index of bone resorption of 10~(-6) mol/L genistein+10 ?g/L IL-1? lied between 10~(-6) mol/L genistein and 10 ?g/L IL-1?.In the organ culture,there was no differencein the content of ACP among all the groups,The index of bone resorption of 10~(-6) mol/L genistein+10?g/L IL-1? was below that of 10?g/L IL-1?,but both were above 1.0.The index of bone resorptionwas below 1.0 in the group of 10~(-6) mol/L genistein.Conclusion:Genistein can suppress obviously thebone resorption simulated by single IL-1?.

Objective To investigate the features of computed tomography (CT) and magnetic resonance imaging (MRI) on pancreatic neuroendocrine tumors (pNENs).Methods The retrospective and descriptive study was adopted.The clinicopathological data of 33 patients with pNENs who were admitted to the Tongji Hospital of Tongji Medical School of Huazhong University of Science and Technology between May 2012 and February 2016 were collected.All the patients underwent plain and enhanced scans of CT and MRI.Observation indicators:(1) overall imaging findings and pathological results of pNENs,(2) imaging findings of functional pNENs,(3) imaging findings of non-functional pNENs.Main analysis indicators included tumor diameter,location,boundary,density,cystic degeneration,enhancement,signal,calcification,with or without pancreaticobiliary duct dilation,with or without surrounding tissues invasion,lymph node and distant organ metastases.Results (1) Overall imaging findings and pathological results of pNENs:of 33 patinets with pNENs,24 underwent CT examination,3 underwent MRI examination and 6 underwent CT and MRI examinations.Tumors of 33 patients were solitary with a diameter of 0.6-16.0 cm.Ten,1,13 and 9 tumors were respectively located at the head of pancreas,uncinate process of pancreas,body of pancreas and tail of pancreas.Thirty-three patients were diagnosed as pNENs by pathological examination,including 20 with functional pNENs (insulinoma) and 13 with non-functional pNENs,and G1,G2 and G3 were respectively detected in 24,7 and 2 patients.The coincidence rate between preoperative CT or MRI examination and pathological examination was 90.9％ (30/33).One,1 and 1 patients were misdiagnosed as pancreatic cancer,enlargement of peripancreatic lymph nodes and duodenal gastrointestinal stromal tumor,respectively.(2) Imaging findings of functional pNENs:tumor diameter of 20 patients with functional pNENs was 0.6-3.0 cm with an average diameter of 1.5 cm.Fòur,10 and 6 tumors were respectively located at the head of pancreas,body of pancreas and tail of pancreas.Of 20 patients with functional pNENs,tumors of 19 patients showed clear boundary and 1 showed unclear boundary,and tumors of 18 patients had uniform density and 2 had uneven density with cystic degeneration,without the occurrence of calcification.Of 20 patients undergoing dynamic enhanced scans,tumors of 19 patients demonstrated obvious enhancement in arterial phase and slightly obvious enhancement or were equal to normal pancreatic tissues in portal vein phase and lag phase,and tumor of 1 patient demonstrated slight enhancement in arterial phase and was equal to or less than normal pancreatic tissues in portal vein phase and lag phase.Tumors in 3 patients undergoing MRI scans were manifested as hypointensity on T1-weighted imaging (T1WI),hyperintensity on T2WI and hyperintensity on DWI (b =1 000 s/m2),with clear imaging.Of 20 patients,1 was accompanied with atrophy of pancreatic tissues at distal tumor,pancreatic duct dilatation,multiple retention cyst and enlargement of lymph nodes around the hepatic artery.(3) Imaging findings of non-functional pNENs:tumor diameter of 13 patients with non-functional pNENs was 1.5-16.0 cm with an average diameter of 5.0 cm.Six,1,3 and 3 tumors were respectively located at the head of pancreas,uncinate process of pancreas,body of pancreas and tail of pancreas.Of 13 patients with non-functional pNENs,tumors of 11 patients showed clear boundary and 2 showed unclear boundary,tumors of 3 patients had uniform density and 10 had uneven density with cystic degeneration,and tumors of 2 patients had calcification.Of 13 patients undergoing dynamic enhanced scans,tumors of 12 patients demonstrated obvious enhancement in arterial phase,continuous enhancement in portal vein phase and lag phase and less obvious enhancement at cystic degeneration area,with marked enlargement of supplying arteries and draining veins in partial tumors.Tumor of 1 patient demonstrated slight enhancement,and its enhancement was slightly less than normal pancreatic tissues in arterial phase,portal vein phase and lag phase,with unclear boundary.Results of MRI scans in 6 patients showed that tumors of 4 patients were manifested as hypointensity on T1WI,slight hyperintensity or mixed signal on T2WI and hyperintensity on DWI (b =1 000 s/m2),and tumors of 2 patients were manifested as hypointensity on T1WI,hypointensity on T2WI and hyperintensity on DWI (b =800 s/m2).Of 13 patients with non-functional pNENs,4 had pancreaticobiliary duct dilation and 7 had local tissues invasion or distant organ metastasis (4 with liver metastasis,1 with peripanereatic lymph node metastasis,1 with liver and peripancreatic lymph node metastases and 1 with liver metastasis combined with splenic venous and arterial invasion),including 1 in G1,4 in G2 and 2 in G3.Of 5 patients with tumor diameter ＞ 5.0 cm,4 were complicated with liver or lymph node metastases.Conclusions CT and MRI features of pNENs have a certain characteristics.For functional pNENs,benign and solid tumor is common,with clear boundary and smaller diameter.For non-functional pNENs,tumor size is bigger and cystic necrosis occurs within the tumor,with various enhancements.

BACKGROUND: Donor complications have been detected following autologous tendon transplantation for posterior cruciate ligament reconstruction. Although artificial tendon development and tissue-engineered tendon have achieved great progresses, there are some issues in clinical application. Since 1980's, allogenic tendon transplantation has aroused increasing attention. OBJECTIVE: To explore the selection of allogenic tendon materials and the effect of their application on reconstructing posterior cruciate ligament. METHODS: A total of 17 patients with posterior cruciate ligament injury of knee joint were treated with cryopreserved allogenic tendon by Tibial-inlay technique. During the operation, two tracts of tendons soaked in gentamicin saline for 15 minutes were conduplicated, and one end of the tendon was cancellous bone screw and fixed to the tibia attachment point of posterior cruciate ligament, and the other end was introduced into the joint through retention suture. The posterior joint capsule was repaired. The patient was placed at supine position, and the knee was flexed for 90°. The other end of the graft was introduced to femoral tunnel, and anterior drawer was tensed, and fixed by screw. RESULTS AND CONCLUSlN: The preoperative posterior drawer test of patients was >2+, including 7 cases of 3+ and 6 of 4+. The postoperative posterior drawer test was 0 in 4 cases, 1+ in 8 cases, 2+ in 4 cases and 3+ in 1 case, suggesting the posterior movement of the knee joint was significantly improved. Lysholm scores of patients were (48.5±4.3) points before operation and (88.3±5.4) points after operation. Results show that cryopreserved allogenic tendon by Tibial-inlay technique could restored function of posterior cruciate ligament with a favorable effect.

Objective To explore the effects of murinecytomegalovirus(MCMV)infected mouse embryonic fibroblasts(MEF)on the proliferation and activation of regulatory T cell in vitro. Methods A co-culture system of T cell and MCMV infected MEF(T-MEF MCMV)was established.The viral load of supernatant was determined by plaque assay.The proliferation of T cells was observed with cell counting.The proportion of CD4+ CD25+ cells was measured by flow cytometry.The levels of Foxp3 protein were measured by Western blot.Reverse transcription polymerase chain reaction(RT-PCR)assay was utilized tO determine whether MCMV infection of MEF influenced the level of transforming growth factor(TGF)-β mRNA.The level of TGF-β protein in supernatant was measured by double-antibody sandwich enzyme linked immunosorbent assay(ELISA).The difference between T-MEF MCMV group and control group was assessed by one-way ANOVA.Results When T cells were co-cultured with MCMV infected MEF for 1 day and 3 days,the viral load in supernatant decreased.But when co-culture lasted for 6 days,the antiviral effect obviously diminished,as the viral load[(5.58±0.67)×105 PFU/mL]of the experimental group showed no statistic difference with MEF MCMV control group[(6.05±0.34)×105PFU/mL].When co-cultured with MCMV infected MEF for 3 days,T cell increased from pre-culture level of[(2.02±0.05)×106/mL]to(2.25± 0.13)×106/mL(P＜0.05).But when co-culture lasted for 6 days,the number of T eelI returned to (2.08±0.14)×106/mL,which had no statistic difference with that of co-culture for 3 days system. Both the expressions of Foxp3 protein and the proportion of CD4+ CD25+ Foxp3+ Treg cells in T-MEF MCMV group were up-regulated as the infection time extended,which were two times and three times of the control group level,respectively.The mRNA level(A value)of TGF-β in MCMV infected MEF increased from baseline of 1.09±0.13 to 3.15±0.54 on day 3 after infection.The expression of TGF-β in supernatant 3 days after infection was(3.85±0.32) μg/L,which was significantly higher than that before infection[(1.74±0.14)μg/L,P＜0.05].Conclusions Activated T cells have antiviral effect.However,the function of T cells is rapidly inhibited after activation,which may be due to the expression of Foxp3 mRNA induced by MCMV infected MEF and increased CD4+ CD25+ Treg proportion of the co-cultured T cells.TGF-β level is significantly increased after CMV infection,which may be an important mechanism of Treg proliferation.MCMV may manipulate Treg to evade specific immune elimination and,as a result,to cause CMV replication.