Objective: To construct mcpr1prokaryoti c expression vector and to express MCPR1 protein.Methods:PCR was used to obtain coding region of mcpr1. Construction of a high-level fusion protein expression vector pGEX-4T-mcpr1 was conducted by inserting the fra gment of coding region of mcpr1into a fusion protein expression vector pGEX -4T-1. Then the recombinant plasmid was transferred into E. colito prepar e the MCPR1/GST fusion protein. DNA sequencing and endonucleases digesting were used to check the coding region. Results:pGEX-4T- mcpr1 wa s constructed successfully and the coding region was inserted into the vector co rrectly. A new protein band of 36 000 was observed by SDS-PAGE analysis after i nduction by IPTG. The 36 000 protein amounted to 39 percent of the total prote in and existed mostly in precipitation of broken bacteria. Conclusion: MCPR1 protein can be expressed in E. coliexpression system and purif ied initially.

Objective To develop a superparamagnetic iron oxide (SPIO)-based MR probe containing vascular endothelial growth factor(VEGF) to investigate their biological and chemical properties and targeting effect of colorectal cancer cells in vitro. Methods The anti-VEGF-SPIO probe was fabricated with VEGF antibody and SPIO through chemical method. Its biological and chemical properties and reflexivity were tested with SDS-PAGE and MRL The SW620 cells incubated with anti-VEGF-SPIO probe for 30, 60 and 90 minutes respectively and compared with marrow mesenchymal stem cell at 37℃. The comparison among groups was conducted by using analysis of variance and LSD-t test. The MRI results were confirmed by the Prussian blue staining. The comparison among groups was performed by analysis of variance and factorial experiment. Results SPIO-based MR probe containing VEGF was successfully contributed and isolated. The reflexivity of anti-VEGF-SPIO probe was 0.0426×10~6 mol/s. The immunofluorescence and prussia blue stain proved high expression of VEGF in SW620 cells. Anti-VEGF-SPIO probe and SW620 cellscombined at 37℃ in vitro MRI proved the SW620 cells incubated with anti-VEGF-SPIO probe appeared hypointense on T_2WI and T_2~* WI. MR signal were 392±7,91±8,264±10 for 30, 60 and 90 minutes respectively, which were statistically different from that before incubation 679±12 (F=4735.489, P< 0.01). The intensity decreased most significantly at 60 minutes in vitro. Its MR signal 82±7 were statistically different compared with marrow mesenchymal stem cell 689±43, t=39.167,P<0.05). While SW620 cells incubated without SPIO were not statistically different compared with marrow mesenchymal stem cell, which were 419±59 and 400±41 respectively(t=-0.718,P>0.05). Conclusion Nanoscale iron particles containing the anti-vascular endothelial growth factor molecular probe can evaluate tumor angiogenesis at the receptor level, which provides a new way of the tumor angiogenesis diagnosis and anti-angiogenesis therapy.

?Advances in genome-wide analysis have revealed that up to 90%of the human genome is transcribed.However, only approximately 1% of RNA transcripts encode proteins, and the remaining transcripts are noncoding RNAs.Noncoding RNAs can be roughly divided into small noncoding RNAs (<200nt ) and long noncoding RNAs ( LncRNAs, >200nt ). Small noncoding RNAs include microRNAs, transfer RNAs and small nucleolar RNAs, whereas the long noncoding RNAs comprise ribosomal RNA, natural antisense transcripts, etc. Although the biosynthesis and biological activities of microRNAs are well studied through bioinformatics and active biological molecules analysis, the understanding of LncRNAs on these aspects is still limited.LncRNAs play multiple roles in regulating gene transcription and translation, and epigenetics.Aberrant LncRNAs expression can occur in various pathological processes and significantly related to the pathogenesis or poor prognosis of ophthalmological diseases. In this review, we will focus on the characteristics and regulatory functions of LncRNAs that are commonly associated with ophthalmological diseases.

Objective To investigate the dosimetric properties of PTW 60019 synthetic diamond detector in small photon beams.Methods A PTW 60019 synthetic diamond detector was tested under 6 and 10 MV photon beams,respectively.Linearity with dose,dose rate dependence and off-axis ratio were measured and compared to those measured by an IBA SFD.Percentage depth doses were measured and compared to those measured by an IBA SFD and a PTW 31010 semiflex chamber.Total scatter factors were measured and compared to those measured by an IBA SFD and a PTW 31016 PinPoint chamber.Results The dose response of a PTW 60019 synthetic diamond detector showed a good linear behavior as a function of dose,with observed deviations below 0.2％ over a dose range from 100 to 1 000 MU.The dose rate response was almost independent,with deviations below 0.2％ in the dose rate range from 37 to 614 MU/min.For the fields of 20-100 mm in diameter,there were dose differences in percentage depth doses within 1％ as compared to an IBA SFD and a PTW 31010 semiflex chamber.For the 10 mm diameter field,the differences were up to 5.8％ in the build-up region.Off-axis ratios measurements showed a good agreement among the involved detectors (＜ 1％).The higher differences appeared in the penumbra region.A good agreement was also found in terms of total scatter factor measurements for the related detectors.Conclusions The observed dosimetric properties of the PTW 60019 synthetic diamond detector indicate that it is a suitable candidate for small photon beam dosimetry.

Objective To investigate the expression of microRNA -216a(miR -216a)in patients suf-fering with breast cancer ,and identify the function and mechanism of miR -216 a on autophagy .Methods The expression of miR-216 a in 30 tumor tissues and paired normal tissue of breast cancer were detected by qRT -PCR.Inhibiton of miR-216a in MCF-7 cell lines was done by transfection of miR -216a inhibitor ( AMO-216a).Cell viability was detected by MTT assay .The level of Beclin 1 was detected by western blot .Results The level of miR-216 a was significant elevated in tumor tissue .Cell viability was markedly decreased owing to inhibition of miR-216a in MCF-7 cells.The level of Beclin 1 was significantly increased by transfection of miR-216a inhibitor in MCF-7cells(P<0.05).Conclusion The level of miR-216a is increased in breast canc-er tissue, and it might at least in part promote breast cancer via downregulating Beclin 1 and affecting autophagy .

Objective To observe the 2-year survival situation of high intensity focused ultrasound(HIFU) treatment in unr esectable advanced pancreatic carcinoma.Methods Thirty-eight patients with unresectable pancreatic cancer received HIFU treatment.After treatment,the changes of laboratory tumor markers examination results,pain score,life quality score and survival situation were recorded.Results Among 35 patients with pain symptom before HIFU treatment,pain was relieved after HIFU treatment in 28 cases,the remission rate was 80.0％.The CA19-9 and CEA levels after HIFU therapy were obviously reduced compared with before treatment.The imaging examination showed the coagulation necrosis in HIFU-treated area.It was found the tumor volume was obviously shrunk during follow-up period.The median survival period was extended to (12.9 ± 6.6) months.Conclusion The HIFU treatment can effectively improve the life quality in the patients with unresectable pancreatic cancer and extends their survival period.

Aim To screen peptides binding specifically to anti-human PTA1mAbs from a random twelve-peptide phage-disp-layed library. Methods Series of PTA1mAbs(LeoA1、 1B11、 C9、 2D1、 2E9、 2G8、 2H2 and E8)were purified using protein-A affinity column. PTA1mAbs which could bind PTA1-Fc fusion protein binding to its ligand were confirmed by flow cytometry, and then used as target to screen phage library. After three rounds of affinity screening, the peptide sequences of positive phage clones were determined and analyzed. Results LeoA1 could block PTA1-Fc fusion protein binding to its ligand. 13 phages which could bind specifically to LeoA1 were isolated from phage library and further confirmed by ELISA. Conserved motifs were found among the sequences of the peptides. Conclusion It was shown that the conserved motifs were candidafe regions binding to PTA1 ligand,which is important to identify functional epitopes for seeking ligand of PTA1 and further investigation of biological function of PTA1.

Objective To determine whether cleavage developmentally retarded embryos have not cleaved during a 24 hour period could develop into blastocysts and produce hESC cell lines.Methods A total of 120 such embryos were cultured to blastocyst stage by sequential culture.Blastocysts formation rate and quality of blastocyst were detected under microscope.The relation between blastocyst formation rate and blastomere number,the fragment of blastomere and blastomere symmetry were analyzed by stepwise Logistical regression analysis.Inner cell masses(ICMs)were isolated by immunosurgery.Colonies derived from the ICMs were passed every 4-7 days and the derivatives were passaged and identified.Results A total of 22 blastocysts were obtained from 120 embryos.The blastulation rate was 18.7%.Early blatocyst, blastocyst,full blastocyst,expanded blastocyst,hatching blastoeyst and hatched blastocyst accounted for 5.9%,23.5%,35.3%,23.5%,5.9%,and 5.9% respectively.The grade of ICM and trophoblast was mostly scored C or B.Blastocyst formation rate was related to cell number and blastomere symmetry but not fragment.Immunosurgery resulted in the formation of 7 ICMs and 3 primary colonies,which produced 2 cell lines.The cell lines satisfied the criteria that characterize pluripotent hESC cells.Undifferentiated cells were positive for AKP,SSEA-4,TRA-1-60,and TRA-1-81.It could continue to proliferate in vitro and form embryoid bodies when cultured in suspension.It had capability to form teratoma in SCID mice.Both cell lines had normal karyotypes after 45 and 34 passages respectively.Conclusions Our results suggest that a subset of developmentally retarded embryos can form blastocysts and give rise to hESC cell lines.

Alpha-glycerophosphate oxidase (alpha-GPO) from Enterococcus casseli flavus was successfully isolated and purified by using polyethylene glycol (PEG)/(NH4)2SO4 aqueous two-phase system (ATPS). The results showed that the chosen PEG/(NH4)2SO4 ATPS could be affected by PEG molecular weight, pH, concentration of PEG and (NH4)2SO4, and inorganic salt as well as additional amount of crude enzyme. After evaluating these influencing factors, the final optimum purification strategy was formed by 16.5% (m/m) PEG2000, 13.2% (m/m) (NH4)2SO4, pH 7.5 and 30% (m/m) additive crude enzyme, respectively. The NaCl was a negative influencing factor which would lead to lower purification fold and activity recovery. These conditions eventually resulted in the activity recovery of 89% (m/m), distribution coefficient of 1.2 and purification fold of 7.0.
Ammonium Sulfate ; chemistry ; Glycerolphosphate Dehydrogenase ; chemistry ; isolation & purification ; Molecular Weight ; Polyethylene Glycols ; chemistry ; Water

Objective To analyze laparoscopic lymphadenectomy and the safety of gastrectomy for gastroesophageal junction cancer.Methods From Jan 2011 to Dec 2012 72 gastroesophageal junction cancer patients were enrolled, including 46 patients in laparoscopic group, and 26 in open surgery group.Results There was no significant difference in the numgbers of lymph node dissection between the two groups and nor difference in the number of positive lymph node dissection.Esophagus resection length in open group was (2.0 ± 1.0) cm, while that was (3.0 ± 0.8) cm in laparoscopic group (t =0.471, P ＜ 0.001).5 (19％)patients in open group had positive margins compared to six in laparoscopic group (13％), x2 =0.491, P =0.483.7 patients in the open group underwent thoracoabdominal resection, while in laparoscopic group 3 patients did, x2 =5.781, P =0.016.Laparoscopic splenic hilar lymphnodes dissection harvested more lymph nodes (t =0.260, P =0.011).Laparoscopic gastrectomy used less operation time (t =0.237, P =0.021) experinced less blood loss (t =0.451, P ＜ 0.01) than open group.There was no difference in major complications between the two groups.Conclusions Laparoscopic splenic hilar lymphnodes dissection in gastroesophageal junction cancer surgery is superior to open surgery, with more lymph nodes harvested, longer esophageal cutting distances, lower incidence of thoracoabdominal surgery, shorter operation time, and less blood loss.