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

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.

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 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.

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.

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 clinical features and microsurgical outcomes of patients with parasellar cavernous hemangiomas.Methods The clinical data and prognosis of 12 patients with parasellar cavemous hemangiomas treated with microsurgery were retrospectively analyzed.Magnetic resonance imaging showed that cavernous hemangiomas appeared slight hypointensity on T1WI,hyperintense on T2WI,and significant contrast on enhancing.Results Total resection and sub-total resection was achieved in 9 and 3 patients.No death occurred postoperatively.After operation,all patients were released from intracranial hypertension.In 6 patients who had visual disorder,5 patients improved and 1 patient had no change.In 5 patients who had eyeball motor disorder,3 patients improved and 2 patients had no change.In 4 patients who had pituitary gland dyshormonism,2 patients improved and 2 patients had no change.At postoperative early stage,diabetes insipidus and serum electrolyte disorder occurred in 3 patients,2 patients developed oculomotor paralysis.Patients were followed up (30.51 + 2.57) months.Three patients recovered from diabetes insipidus and serum electrolyte disorder,and 1 patient recovered from oculomotor paralysis.No patient suffered tumor recurrence or regrowth.Conclusions The correct diagnosis of parasellar cavernous hemangiomas can be achieved according to the preoperative MRI.Microsurgery is the first-line therapy for parasellar cavernous hemangiomas.

BACKGROUND:Skeletal muscle satel ite cells are totipotential stem cells with multi-directional differentiation potential, locate in skeletal muscle interstitium, have a certain tolerance to ischemia and hypoxia, and are important cells in stem cellengineering.
OBJECTIVE:To establish a thrifty, convenient culture procedure and create a simple, efficient method to transfect skeletal muscle satel ite cells, and investigate genetic expression after the transfection for cellular cardiomyoplasty.
METHODS:Skeletal muscle satel ite cells were isolated from rabbit thigh and cultured. Their growth curves were determined by CKK-8 method. Grouped by different proportions of the plasmid and liposome, skeletal muscle satel ite cells were transfered by the enhanced green fluorescent protein plasmid based on liposome. After transfection, the efficiency and character of target genetic expression was determined.
RESULTS AND CONCLUSION:Satel ite cells were isolated, cultured and transfected successful y. In suitable ratio of plasmid and liposomes, the transfection efficiency reached up to above 35%. The target protein was expressed within 12 hours after transfection, reached maximum in 48-72 hours and decreased gradual y after one week. The expression stil could be observed two weeks latter. The enhanced green fluorescent protein plasmid conducted by cationic liposome could be transfered into skeletal muscle satel ite cells efficiently. The transfection efficiency was correlated closely to the ratio of plasmid and lipofectamine. The change of target gene expression depended on time.

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.

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.