Objective To evaluate the efficiency of primary hepatic carcinoma(PHC)using tran- scatheter arterial chemoembolization(TALE)combining with MSC.Methods Treatment group(30 patients with unresectable PHC)adopt MSC after TALE,while control group(10 patients with unresectable PHC)only adopt TALE,then observing the curative effect and survival rate.Results One year after treating,the local effect was 90 %,60 %,1～2 year survival rate was 90 %,30 % respectively,median life span was 15 months. Conclusion To treat PHC with TALE combining with MSC is an effective and non-invasive method.

Background Choroidal neovascularization (CNV) is a serious complication of many fundus diseases.A variety of factors are associated with CNV.Research showed that recombinant human endostatin ( rhendostatin ) can arrest the vascular endothelial growth factor (VEGF) pathway and inhibit the proliferation of endothelial cells and angiogenesis.Objective This study was to observe the inhibition of rh-endostatin on experimental CNV.Methods The CNV animal models were created by Argon laser with the wavelength 532 nm to irradiate the inferior retina away optical disc 1-2 DD for 25 spots in 32 eyes of 16 chinchilla rabbits.The laser parameters were as follows:power 800 mW,spot diameter 75 μm and time shutter 50 ms.The models were then divided into model control group and rh-endostatin group.Rh-endostatin was intravitreously injected via scleral incision in 16 eyes of 8 model rabbits at 1 week after photocoagulation.Fundus fluorescein angiography (FFA) and optical coherence topography(OCT) were performed at 1,2,4 weeks after photocoagulation respectively.The eyeballs were enucleated and the retinal sections were prepared for the histopathologieal examination,and the contents of VEGF and pigment epithelial derived factor(PEDF) in rabbit vitreous,and blood serum were detected by ELISA at 2,4 weeks after photocoagulation.Results Retinal edema and exudes were seen in 1 week and scarring in 4 weeks after photocoagulation.In rh-endostatin injection group,the hyperfluorescence masses were seen in the background phase and early arterial phase in 42％ (84/200) of spots in the first week.The fluorescence leakage was decreased in the rh-endostatin injection group compared with control group in the second week and ceased at the third week on the FFA after injection.Variety forms of hyperreflective zones were found below the retinal pigment epithelium on the seventh day after photocoagulation.But the partial vessel occlusion and fibroplasias were identified in the rh-endostatin injection group in the third week by the OCT.The histopathological examination showed that the morphological abnormality was mild in the rh-endostatin injection group in comparison with model group.The serum PEDF concentration was significantly elevated but the VEGF/PEDF values in vitreous and serum were declined in rhendostatin injection group compared with model group (P ＜ 0.0 1 ).Conclusions Argon laser photocoagulation could induce the experimental CNV in chinchilla rabbit.Intravitreous injection of rh-endostar can effectively inhibit laser-induced CNV in rabbit.

Objective To study the influence of low-iodine diet on the expression of homeobox gene nkx2.1 in rat cerebral tissue. Methods Forty female Wistar rats were randomly divided into two groups according to body. quality: low-iodine group and control group,both fed with low-iodine feed at an iodine content of 13.66 μg/kg,respectively given the deionized water and 200 μg/L KIO3 solution. The hormone levels of two group rats were determined with chemiluminescence immunoassay after three months, and then mated with healthy male rats. Cerebral tissues were taken from the fetus of 16-day pregnancy,newborn and 20 days old offspring in low-iodine and control group to detect the content of nkx2.1 mRNA using real-time fluorescence quantitative PCR (FQ-PCR) techniques. Results Serum TT3, TT4, FT3, FT4 level of rats in low-iodine group(0.89±0.20, 0.32±0.16, 3.33± 0.61, 3.28±0.80) was respectively lower than that in the control group(1.04±0.06, 39.42±14.68,4.83±0.33, 26.99±4.48;t = 2.71,6.52,5.70, 12.89, P < 0.05 or < 0.01). The relative nkx2.1 mRNA expression was(5.60± 0.30)×10-3, (1.20 ± 0.29)×10-3, (0.18± 0.06)×10-3 respectively in the fetus of 16-day pregnancy, newborn and 20 days old offspring of control group, while it was (3.00 ± 0.55)×10-3, (1.90 ± 0.21)×10-3,(0.69 ± 0.15)×10-3 in the low-iodine group. The difference of nkx2.1 mRNA expression was significant among fetal and neonatal rats in the control group and low-iodine group(F = 210.07,162.40, both P < 0.01). The nkx2.1 mRNA expression of newborn rats was lower than that of 16-day pregnancy in both groups(P < 0.01), and that of 20 days old rats was lower than that of 16-day pregnant and neonatal rats(P < 0.01). The 16-day pregnant rats of control group had obviously higher level of nkx2.1 expression than those in the low-iodine group(t = 16.073, P< 0.01), while the nkx2.1 of newborn and 20 days old low-iodine rats expressed much higher than healthy rats(t = 7.573,12.221, P < 0.01). Conclusions Brain development retardation caused by low-iodine is closely related to nkx2.1 differential expression in the brain tissue.

Objective To study the influence of low-iodine diet on the expression of homeobox gene Nkx-6.1 and Nkx-6.2 in rat cerebrum tissue, and to explore the possible molecular mechanism of cerebrum development retardation caused by low-iodine. Methods Twenty female Wistar rats were randomly equally divided into two groups: low-iodine group and control group, both fed with low-iodine diet as low as 13.66 μg/kg determinated by spectrophotometry in Tianjin Institute of Endocrinology and the former with deionized water, the later 200 μg/L potassium iodate. Thyroid hormone level was detected using chemiluminescence immunoassay 3 months later and they were mated with male rats normally fed. Rats of 16-day pregnancy, new-born and 20th days old were detected the content of Nkx-6.1 and Nkx-6.2 mRNA in the cerebrum tissue by real-time fluorescence quantitative PCR 0.61), (3.28±0.80)pmol/L] were lower than the control group[(1.04±0.06), (39.42±14.68)nmol/L, (4.83±0.33), day pregnancy, new-born and 20th days old of control group was (1.90±0.23)×10-3,(1.86±0.40)×10-4, (1.11± 0.27)×10-4(F=827.58, P<0.01), Nkx-6.1 mRNA expression level gradually decreased along with aging(all P<0.05). The intra-group difference was significant (F=297.25, P<0.01) and the Nkxr.1 mRNA expression level during 16 days of pregnancy was the highest(P<0.01). It was higher in the control group than in the low-iodine group during 16 days of pregnancy (t=10.14, P<0.01) as well as in the low-iodine group than in the in 16-day pregnancy, new-born and 20th days old of control group was respectively(1.03±0.19)×10-2, (1.33± 0.10)×10-3, (8.79±0,87)×10-3, and that of low-iodine group was (0.31±0.03)×10-2, (1.53±0.13)×10-3, (7.51±0.86)×10-2. The intra-group difference was significant(F=1293.02,1065.83, all P<0.01). Nkx-6.2 expression level during 20th days old was the highest(P<0.01) and that of newborn was the lowest(P<0.01). The Nkx6.2 mRNA expression level in control group were higher than the low-iodine group during 16-day pregnancy and 20th days old(t=14.35, 4.05, all P<0.01). It was higher in the low-iodine group than in the control group during newboru(t=4.78, P<0.01). Conclusions The difference in the expression of Nkx-6.1 and Nkx-62 is highly related to the brain development retardation caused by low-iodine.

Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

OBJECTIVE: To eludicate the risk factors of mechanical ventilation and prolonged mechanical ventilation in patients with severe multiple injuries. METHODS: Consecutive patients with severe multiple injures who were treated in Peking University People's Hospital Trauma Medical Center between December 2016 and December 2019 were enrolled in this restropective chart-review study. According to mechanical ventilation and ventilatory time, the patients were divided into mechanical ventilation (MV) group and non-mechanical ventilation (NMV) groups, prolonged mechanical ventilation (PMV) group and shortened mechanical ventilation (SMV) groups. Clinical data such as gender, age, base excess, mechanism of injury, Glasgow Coma Scale (GCS), abbreviated injury scale (AIS) and injury severity score (ISS) were collected. To indentify the risk factors of mechanical ventilation and prolonged mecha-nical ventilation, univariate and multivariate Logistic analyses were carried out. RESULTS: In the present study, 112 patients (82 male, 30 female) with severe multiple injuries having a median age of 52 (range: 16-89 years) and a median ISS of 34 (range: 16-66) were enrolled. The primary mechanism of injury was traffic accident injury and falling injury. In the study, 62 and 50 patients were assigned to MV and NMV groups, respectively. Logistic analysis showed that GCS (OR=0.72, 95%CI: 0.53-0.92, P=0.03), base excess (OR=0.56, 95%CI: 0.37-0.88, P=0.002) and multiple rib fracture (OR=1.72, 95%CI: 1.60-2.80, P=0.012) were independent significant risk factors for mechanical ventilation after severe multiple injuries. Within the mechanical ventilation group, 38 and 24 patients were assigned to PMV and SMVgroups, respectively. Compared with the SMV group, the PMV group had a higher ISS and higher rate of severe head trauma. The length of hospital stay of PMV group was longer than that of SMV groups. Meanwhile, the incidence of tracheotomy in PMV group was high. CONCLUSIONS GCS, base excess and rib fracture might be independent risk factors for mechanical ventilation. Higher ISS and lower GCS might prolong the ventilatory time and the length of hospital stay. Meanwhile, the incidence of tracheotomy was high in PMV group because of the longer ventilatory time and poor consciousness.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Injury Severity Score ; Male ; Middle Aged ; Multiple Trauma ; Respiration, Artificial ; Retrospective Studies ; Risk Factors ; Young Adult

OBJECTIVESTo investigate whether antisense Bmi-1 plasmid could inhibit the proliferation of Jurkat cells.

METHODSThe antisense plasmid was constructed by PCR amplification of a 171 bp segment spanning Bmi-1 start codon and zinc finger structure and the PCR product was subsequently inserted reversely to plasmid pLNCX2. The final construct was confirmed through restriction enzyme digestion. G418 was added into the medium after the plasmid was successfully introduced into Jurkat cells by using lipofectin-mediated DNA transfection. The proliferation of Jurkat cells were determined by MTT and colony formation assays. Cell cycle was determined by flow cytometry. The p16 expression of Jurkat cells was studied by immunofluorescent histochemistry.

RESULTSThe growth rate of antisense Bmi-1 transfected Jurkat cells was significantly lower than that of the controls, and the colony forming capacity of the transfected cells decreased significantly (P < 0.01), the colony numbers being (90.7 +/- 9.07)/10(3) cells, (83.3 +/- 6.11)/10(3) cells and (56.0 +/- 5.56)/10(3) cells for control cells, empty plasmid transfected Jurkat cells and antisense Bmi-1 transfected Jurkat cells, respectively. The percentage of G, phase cells was increased and the p16 expression of antisense Bmi-1 transfected cells was significantly upregulated than that of control cells.

CONCLUSIONAntisense Bmi-1 can inhibit the growth and upregulate the expression of p16 of Jurkat cells in vitro.

Cell Cycle ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Jurkat Cells ; Nuclear Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Plasmids ; genetics ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; Transfection

To clone the 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (TwMCT) full length cDNA from Tripterygium wilfordii, the specific primers were designed according to the transcriptome data and the LCPCR were carried out. After a series of bioinformatics analysis on the TwMCT, the MeJA induced expression content were investigated by real-time fluorescence quantification polymerase chain reaction (RT-qPCR). The result showed that the full of TwMCTcDNA was 1 318 bp nucleotides encoding 311 amino acids. The molecular weight of the deduced TwMCT protein was about 34.14 kDa and the theoretical isoelectric point was 8.65. Result of the RT-qPCR analysis indicated that the content of TwMCT mRNA expression in T. wilfordii suspension cell was rising after treating with MeJA and reached the maximum in 24 h. Cloning and analyzing TwMCT gene from T. wilfordii provided gene element for studying the function and expression regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Erythritol ; analogs & derivatives ; metabolism ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Nucleotidyltransferases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Sequence Alignment ; Sugar Phosphates ; metabolism ; Tripterygium ; chemistry ; enzymology ; genetics

Albuminuria ; drug therapy ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; Diabetic Nephropathies ; drug therapy ; Humans ; Tetrazoles ; therapeutic use

ObjectiveTo investigate the value of 99Tcm labeled survivin mRNA antisense peptide nucleic acid (PNA) as an imaging agent in the specific diagnosis for carcinoma.MethodsSurvivin mRNA antisense PNA was labeled directly with 99Tcm by the ligand-exchange method.Twenty nude mice with lung carcinoma A549 xenografts were randomly divided into 4 groups.Three groups were used for biodistribution study and one group was used for imaging study.Other twenty mice infected by staphylococcus aureus underwent the same procedure.The biodistribution and imaging of 99Tcm-survivin mRNA antisense PNA was studied at 1,2 and 4 h respectively after the intravenous injection in nude mice bearing lung carcinoma A549 xenografts or inflammation models.SPSS 13.0 was used in the study and all data were analyzed by t test.ResultsBiodistribution results showed that the highest radioactivity was found in the liver,and then in the kidney.Four hours after the administration of the imaging agent,the radioactivity ratios of target-tonon target (T/NT,tumor or inflamumatory lesions to the contralateral regions) in tumor model group were significantly higher than those in inflammation model group ( 3.69 ± 1.13 vs 2.03 ± 0.47,t =3.01,P =0.02 ).Tumors were clearly visible in the tumor model groups at 0.5 h and still clearly seen at 4 h after the injection of antisense PNA.On the contrary,inflammatory lesions could not be seen clearly.Conclusion 99Tcm labeled survivin mRNA antisense PNA can be used to distinguish tumor from inflammation and it may provide a new feasible method for specific tumor diagnosis.